@article {pmid38535401, year = {2024}, author = {Cholvi, M and Trelis, M and Bueno-Marí, R and Khoubbane, M and Gil, R and Marcilla, A and Moretti, R}, title = {Wolbachia Infection through Hybridization to Enhance an Incompatible Insect Technique-Based Suppression of Aedes albopictus in Eastern Spain.}, journal = {Insects}, volume = {15}, number = {3}, pages = {}, doi = {10.3390/insects15030206}, pmid = {38535401}, issn = {2075-4450}, abstract = {The emergence of insecticide resistance in arbovirus vectors is putting the focus on the development of new strategies for control. In this regard, the exploitation of Wolbachia endosymbionts is receiving increasing attention due to its demonstrated effectiveness in reducing the vectorial capacity of Aedes mosquitoes. Here, we describe the establishment of a naïve Wolbachia infection in a wild Aedes albopictus population of eastern Spain through a hybridization approach to obtain males capable of sterilizing wild females. The obtained lines were compared with the Wolbachia donor, Ae. albopictus ARwP, previously artificially infected with Wolbachia wPip, regarding immature and adult survival, female fecundity, egg fertility, and level of induced sterility. Our results did not show significant differences between lines in any of the biological parameters analyzed, indicating the full suitability of the hybrids to be used as a control tool against Ae. albopictus. In particular, hybrid males induced 99.9% sterility in the eggs of wild females without the need for any preliminary treatment. Being harmless to non-target organisms and the environment, the use of this bacterium for the control of Ae. albopictus deserves further exploration. This is especially relevant in areas such as eastern Spain, where this mosquito species has recently spread and may represent a serious threat due to its competence as a vector for dengue, chikungunya, and Zika viruses.}, } @article {pmid38534451, year = {2024}, author = {Miranda, LS and Rudd, SR and Mena, O and Hudspeth, PE and Barboza-Corona, JE and Park, HW and Bideshi, DK}, title = {The Perpetual Vector Mosquito Threat and Its Eco-Friendly Nemeses.}, journal = {Biology}, volume = {13}, number = {3}, pages = {}, doi = {10.3390/biology13030182}, pmid = {38534451}, issn = {2079-7737}, support = {na//an Internal Research Grant through the Office of the Provost and the Faculty Development Committee at California Baptist University./ ; }, abstract = {Mosquitoes are the most notorious arthropod vectors of viral and parasitic diseases for which approximately half the world's population, ~4,000,000,000, is at risk. Integrated pest management programs (IPMPs) have achieved some success in mitigating the regional transmission and persistence of these diseases. However, as many vector-borne diseases remain pervasive, it is obvious that IPMP successes have not been absolute in eradicating the threat imposed by mosquitoes. Moreover, the expanding mosquito geographic ranges caused by factors related to climate change and globalization (travel, trade, and migration), and the evolution of resistance to synthetic pesticides, present ongoing challenges to reducing or eliminating the local and global burden of these diseases, especially in economically and medically disadvantaged societies. Abatement strategies include the control of vector populations with synthetic pesticides and eco-friendly technologies. These "green" technologies include SIT, IIT, RIDL, CRISPR/Cas9 gene drive, and biological control that specifically targets the aquatic larval stages of mosquitoes. Regarding the latter, the most effective continues to be the widespread use of Lysinibacillus sphaericus (Ls) and Bacillus thuringiensis subsp. israelensis (Bti). Here, we present a review of the health issues elicited by vector mosquitoes, control strategies, and lastly, focus on the biology of Ls and Bti, with an emphasis on the latter, to which no resistance has been observed in the field.}, } @article {pmid38534421, year = {2024}, author = {Hyder, M and Lodhi, AM and Wang, Z and Bukero, A and Gao, J and Mao, R}, title = {Wolbachia Interactions with Diverse Insect Hosts: From Reproductive Modulations to Sustainable Pest Management Strategies.}, journal = {Biology}, volume = {13}, number = {3}, pages = {}, doi = {10.3390/biology13030151}, pmid = {38534421}, issn = {2079-7737}, support = {32202276//National Science Foundation of China/ ; 2022GDASZH-2022010106, 2022GDASZH-2022030501-08//GDAS Special Project of Science and Technology Development/ ; KTP20210352//Guangdong Province Rural Science and Technology Commissioner Project/ ; 2023SDZG06//Top Ten Critical Priorities of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province/ ; }, abstract = {Effective in a variety of insect orders, including dipteran, lepidopteran, and hemipteran, Wolbachia-based control tactics are investigated, noting the importance of sterile and incompatible insect techniques. Encouraging approaches for controlling Aedes mosquitoes are necessary, as demonstrated by the evaluation of a new SIT/IIT combination and the incorporation of SIT into Drosophila suzukii management. For example, Wolbachia may protect plants from rice pests, demonstrating its potential for agricultural biological vector management. Maternal transmission and cytoplasmic incompatibility dynamics are explored, while Wolbachia phenotypic impacts on mosquito and rice pest management are examined. The importance of host evolutionary distance is emphasised in recent scale insect research that addresses host-shifting. Using greater information, a suggested method for comprehending Wolbachia host variations in various contexts emphasises ecological connectivity. Endosymbionts passed on maternally in nematodes and arthropods, Wolbachia are widely distributed around the world and have evolved both mutualistic and parasitic traits. Wolbachia is positioned as a paradigm for microbial symbiosis due to advancements in multiomics, gene functional assays, and its effect on human health. The challenges and opportunities facing Wolbachia research include scale issues, ecological implications, ethical conundrums, and the possibility of customising strains through genetic engineering. It is thought that cooperative efforts are required to include Wolbachia-based therapies into pest management techniques while ensuring responsible and sustainable ways.}, } @article {pmid38530785, year = {2024}, author = {Fricke, LC and Lindsey, ARI}, title = {Identification of parthenogenesis-inducing effector proteins in Wolbachia.}, journal = {Genome biology and evolution}, volume = {}, number = {}, pages = {}, doi = {10.1093/gbe/evae036}, pmid = {38530785}, issn = {1759-6653}, abstract = {Bacteria in the genus Wolbachia have evolved numerous strategies to manipulate arthropod sex, including the conversion of would-be male offspring to asexually reproducing females. This so-called "parthenogenesis-induction" phenotype can be found in a number of Wolbachia strains that infect arthropods with haplodiploid sex determination systems, including parasitoid wasps. Despite the discovery of microbe-mediated parthenogenesis more than 30 years ago, the underlying genetic mechanisms have remained elusive. We used a suite of genomic, computational, and molecular tools to identify and characterize two proteins that are uniquely found in parthenogenesis-inducing Wolbachia and have strong signatures of host-associated bacterial effector proteins. These putative parthenogenesis-inducing proteins have structural homology to eukaryotic protein domains including nucleoporins, the key insect sex-determining factor Transformer, and a eukaryotic-like serine-threonine kinase with leucine rich repeats. Furthermore, these proteins significantly impact eukaryotic cell biology in the model Saccharomyces cerevisiae. We suggest these proteins are parthenogenesis-inducing factors and our results indicate this would be made possible by a novel mechanism of bacterial-host interaction.}, } @article {pmid38528862, year = {2024}, author = {Soo, WF and Gunasekaran, K and Ng, DX and Kwek, K and Tan, NC}, title = {Literacy and attitude of Asian youths on dengue and its prevention in an endemic developed community.}, journal = {Frontiers in public health}, volume = {12}, number = {}, pages = {1361717}, pmid = {38528862}, issn = {2296-2565}, abstract = {BACKGROUND: Over the past few decades, the incidence of dengue fever has considerably increased. Effective vector control strategies and specific protection using dengue vaccine are thought to be the key elements to combat dengue. The dengue incidence among the Singapore youths (15-24 years) was second only to that of adults (25-44 years). This study evaluated the knowledge and attitude of Singapore youths on dengue and its preventive measures.

METHODS: A cross-sectional study using online-based questionnaire survey was conducted among Singapore youths from September to November 2022. Data were analyzed for descriptive statistics whereas Chi-squared test, linear regression analysis and Pearson correlation were used to determine the association between demographic factors and youth's attitude on dengue prevention using Rstudio.

RESULTS: A total of 624 respondents completed the survey out of 1822 surveys distributed nation-wide, with a response rate of 34.2% (mean age 17.4 years ± 1.84; 59.3% female; 89.9% Chinese). The mean dengue knowledge scores of participants were 14.1 ±2.8. Univariate analysis showed that teenagers (15-19 years) had significantly higher knowledge score than the young adults (20-24 years) (β=0.82,95%CI = 0.13-1.51, p = 0.021). Majority of them were aware of the Mozzie Wipeout campaign (90.2%) followed by the release of Wolbachia mosquitos (69.1%). Two-thirds of the youths who were aware of Wolbachia and Gravitrap considered that it was effective in reducing dengue infection rates. Participants suggested information about the current dengue infection rate (71.9%) as the most effective of the five proposed strategies to improve uptake of dengue preventive measures. In comparison to young adults, teenagers were more likely to uptake dengue preventive measures if widespread mosquito control practices were implemented (69.1% vs. 42.3%).

CONCLUSION: The overall knowledge of the youths on dengue and its prevention was satisfactory. Future health promotion campaigns targeting the youths should focus on transforming the knowledge into practice.}, } @article {pmid38525276, year = {2024}, author = {Guse, K and Pietri, JE}, title = {Endosymbiont and gut bacterial communities of the brown-banded cockroach, Supella longipalpa.}, journal = {PeerJ}, volume = {12}, number = {}, pages = {e17095}, pmid = {38525276}, issn = {2167-8359}, abstract = {The brown-banded cockroach (Supella longipalpa) is a widespread nuisance and public health pest. Like the German cockroach (Blattella germanica), this species is adapted to the indoor biome and completes the entirety of its life cycle in human-built structures. Recently, understanding the contributions of commensal and symbiotic microbes to the biology of cockroach pests, as well as the applications of targeting these microbes for pest control, have garnered significant scientific interest. However, relative to B. germanica, the biology of S. longipalpa, including its microbial associations, is understudied. Therefore, the goal of the present study was to quantitatively examine and characterize both the endosymbiont and gut bacterial communities of S. longipalpa for the first time. To do so, bacterial 16S rRNA gene amplicon sequencing was conducted on DNA extracts from whole adult females and males, early instar nymphs, and late instar nymphs. The results demonstrate that the gut microbiome is dominated by two genera of bacteria known to have beneficial probiotic effects in other organisms, namely Lactobacillus and Akkermansia. Furthermore, our data show a significant effect of nymphal development on diversity and variation in the gut microbiome. Lastly, we reveal significant negative correlations between the two intracellular endosymbionts, Blattabacterium and Wolbachia, as well as between Blattabacterium and the gut microbiome, suggesting that Blattabacterium endosymbionts could directly or indirectly influence the composition of other bacterial populations. These findings have implications for understanding the adaptation of S. longipalpa to the indoor biome, its divergence from other indoor cockroach pest species such as B. germanica, the development of novel control approaches that target the microbiome, and fundamental insect-microbe interactions more broadly.}, } @article {pmid38507220, year = {2024}, author = {Hollingsworth, BD and Cho, C and Vella, M and Roh, H and Sass, J and Lloyd, AL and Brown, ZS}, title = {Economic Optimization of Wolbachia-infected Aedes aegypti release to prevent dengue.}, journal = {Pest management science}, volume = {}, number = {}, pages = {}, doi = {10.1002/ps.8086}, pmid = {38507220}, issn = {1526-4998}, abstract = {BACKGROUND: Dengue virus, primarily transmitted by the Aedes aegypti mosquito, is a major public health concern affecting approximately 3.83 billion people worldwide. Recent releases of Wolbachia-transinfected Ae. aegypti in several cities worldwide have shown that it can reduce dengue transmission. However, these releases are costly, and, to date, no framework has been proposed for determining economically optimal release strategies that account for both costs associated with disease risk and releases.

RESULTS: We present a flexible stochastic dynamic programming framework for determining optimal release schedules for Wolbachia-transinfected mosquitoes that balances the cost of dengue infection with the costs of rearing and releasing transinfected mosquitoes. Using an ordinary differential equation model of Wolbachia and dengue in a hypothetical city loosely describing areas at risk of new dengue epidemics, we determined that an all-or-nothing release strategy that quickly brings Wolbachia to fixation is often the optimal solution. Based on this, we examined the optimal facility size, finding that it was less than unitary elastic with respect to the mosquito population size, with a 100% increase in population size resulting in a 50-67% increase in optimal facility size. Further, we found that these results are robust to mosquito life-history parameters and are mostly determined by the mosquito population size and the fitness costs associated with Wolbachia.

CONCLUSIONS: These results reinforce that Wolbachia-transinfected mosquitoes can reduce the cost of dengue epidemics. Further, they emphasize the importance of determining the size of the target population and fitness costs associated with Wolbachia before releases occur. This article is protected by copyright. All rights reserved.}, } @article {pmid38504126, year = {2024}, author = {Du Toit, A}, title = {Drosophila sperm sabotage by Wolbachia prophage.}, journal = {Nature reviews. Microbiology}, volume = {}, number = {}, pages = {}, pmid = {38504126}, issn = {1740-1534}, } @article {pmid38502456, year = {2024}, author = {Singh, AS and Pathak, D and Devi, MS and Anifowoshe, AT and Nongthomba, U}, title = {Antibiotic alters host's gut microbiota, fertility, and antimicrobial peptide gene expression vis-à-vis ampicillin treatment on model organism Drosophila melanogaster.}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {}, number = {}, pages = {}, pmid = {38502456}, issn = {1618-1905}, support = {DBT-RA/2022/January NE/994//Department of Biotechnology, Government of India./ ; DBT-RA/2022/January NE/994//Department of Biotechnology, Government of India./ ; }, abstract = {Antibiotics are commonly used to treat infectious diseases; however, persistence is often expressed by the pathogenic bacteria and their long-term relative effect on the host have been neglected. The present study investigated the impact of antibiotics in gut microbiota (GM) and metabolism of host. The effect of ampicillin antibiotics on GM of Drosophila melanogaster was analyzed through deep sequencing of 16S rRNA amplicon gene. The dominant phyla consisted of Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria, Planctomycetes, Chloroflexi, Euryarchaeota, Acedobacteria, Verrucomicrobia, and Cyanobacteria. It was found that the composition of GM was significantly altered on administration of antibiotics. On antibiotic treatments, there were decline in relative abundance of Proteobacteria and Firmicutes, while there were increase in relative abundance of Chlorophyta and Bacteroidota. High abundance of 14 genera, viz., Wolbachia, Lactobacillus, Bacillus, Pseudomonas, Thiolamprovum, Pseudoalteromonas, Vibrio, Romboutsia, Staphylococcus, Alteromonas, Clostridium, Lysinibacillus, Litoricola, and Cellulophaga were significant (p ≤ 0.05) upon antibiotic treatment. Particularly, the abundance of Acetobacter was significantly (p ≤ 0.05) declined but increased for Wolbachia. Further, a significant (p ≤ 0.05) increase in Wolbachia endosymbiont of D. melanogaster, Wolbachia endosymbiont of Curculio okumai, and Wolbachia pipientis and a decrease in the Acinetobacter sp. were observed. We observed an increase in functional capacity for biosynthesis of certain nucleotides and the enzyme activities. Further, the decrease in antimicrobial peptide production in the treated group and potential effects on the host's defense mechanisms were observed. This study helps shed light on an often-overlooked dimension, namely the persistence of antibiotics' effects on the host.}, } @article {pmid38496649, year = {2024}, author = {Hague, MTJ and Wheeler, TB and Cooper, BS}, title = {Comparative analysis of Wolbachia maternal transmission and localization in host ovaries.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2024.03.03.583170}, pmid = {38496649}, abstract = {Many insects and other animals carry microbial endosymbionts that influence their reproduction and fitness. These relationships only persist if endosymbionts are reliably transmitted from one host generation to the next. Wolbachia are maternally transmitted endosymbionts found in most insect species, but transmission rates can vary across environments. Maternal transmission of w Mel Wolbachia depends on temperature in natural Drosophila melanogaster hosts and in transinfected Aedes aegypti , where w Mel is used to block pathogens that cause human disease. In D. melanogaster , w Mel transmission declines in the cold as Wolbachia become less abundant in host ovaries and at the posterior pole plasm (the site of germline formation) in mature oocytes. Here, we assess how temperature affects maternal transmission and underlying patterns of Wolbachia localization across 10 Wolbachia strains diverged up to 50 million years-including strains closely related to w Mel-and their natural Drosophila hosts. Many Wolbachia maintain high transmission rates across temperatures, despite highly variable (and sometimes low) levels of Wolbachia in the ovaries and at the developing germline in late-stage oocytes. Identifying strains like closely related w Mel-like Wolbachia with stable transmission across variable environmental conditions may improve the efficacy of Wolbachia -based biocontrol efforts as they expand into globally diverse environments.}, } @article {pmid38493661, year = {2024}, author = {González, MA and Ruiz-Arrondo, I and Bravo-Barriga, D and Cervera-Acedo, C and Santibáñez, P and Oteo, JA and Miranda, MÁ and Barceló, C}, title = {Surveillance and screening of Stomoxyinae flies from Mallorca Island (Spain) reveal the absence of selected pathogens but confirm the presence of the endosymbiotic bacterium Wolbachia pipientis.}, journal = {Research in veterinary science}, volume = {171}, number = {}, pages = {105206}, doi = {10.1016/j.rvsc.2024.105206}, pmid = {38493661}, issn = {1532-2661}, abstract = {Adult brachycera biting flies can significantly impact livestock through both direct effects (reduction of food intake, disturbance, painful bites, and blood loss) and indirect effects (pathogen transmission), leading to substantial economic losses and production damage. This study aimed to assess the presence of blood-sucking flies in six mixed-animal farm environments on the island of Mallorca (Balearic Islands, Spain) by employing multiple trapping methods. Additionally, distribution maps of brachycera biting fly species recorded in Spain were created, based on data extracted thorough review of scientific literature and citizen digital databases. Investigation of several pathogens, including equine infectious anemia virus (EIAV), Anaplasmataceae bacteria, and piroplasm protozoa, was carried out using different PCR targets (18S rRNA, 16S rRNA, groESL, and tat genes). Citizen science databases and literature review corroborated the consistent distribution trend for two Stomoxyinae species, underscoring the importance of citizen collaboration as a complement to traditional entomological surveillance. Our study confirmed the presence of two biting Stomoxyinae species: the prevalent stable fly Stomoxys calcitrans across all sampled farms, and the horn fly Haematobia irritans, which turned out to be less abundant. DNA barcoding techniques validated the identification of the two species. Neither EIAV nor bacterial/protozoan pathogens were detected using the selected PCR targets in either fly species. However, Wolbachia pipientis (clustered in the supergroup A together with the only sequence of W. pipientis from the USA) was identified through PCR targeting 16S rRNA, groESL and wsp genes in all pools of H. irritans (n = 13) collected from two of the examined farms. This study represents the first attempt to investigate pathogens in Stomoxyinae biting flies in Spain. The discovery of the endosymbiotic Wolbachia organism in H. irritans represents the first record in Spain and the second from Europe. This finding holds significant implications for future research on the applications of this bacterium in biocontrol programs.}, } @article {pmid38489098, year = {2024}, author = {Calleros-González, P and Ibarra-Juarez, A and Lamelas, A and Suárez-Moo, P}, title = {How host species and body part determine the microbial communities of five ambrosia beetle species.}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {}, number = {}, pages = {}, pmid = {38489098}, issn = {1618-1905}, abstract = {The ambrosia beetles are farming insects that feed mainly on their cultivated fungi, which in some occasions are pathogens from forest and fruit trees. We used a culture-independent approach based on 16S and 18S rRNA gene metabarcoding analysis to investigate the diversity and composition of the bacterial and fungal communities associated with five ambrosia beetle species: four species native to America (Monarthrum dimidiatum, Dryocoetoides capucinus, Euwallacea discretus, Corthylus consimilis) and an introduced species (Xylosandrus morigerus). For the bacterial community, the beetle species hosted a broad diversity with 1,579 amplicon sequence variants (ASVs) and 66 genera, while for the fungal community they hosted 288 ASVs and 39 genera. Some microbial groups dominated the community within a host species or a body part (Wolbachia in the head-thorax of E. discretus; Ambrosiella in the head-thorax and abdomen of X. morigerus). The taxonomic composition and structure of the microbial communities appeared to differ between beetle species; this was supported by beta-diversity analysis, which indicated that bacterial and fungal communities were clustered mainly by host species. This study characterizes for the first time the microbial communities associated with unexplored ambrosia beetle species, as well as the factors that affect the composition and taxonomic diversity per se, contributing to the knowledge of the ambrosia beetle system.}, } @article {pmid38484041, year = {2024}, author = {Simmons, CP and Donald, W and Tagavi, L and Tarivonda, L and Quai, T and Tavoa, R and Noran, T and Manikaoti, E and Kareaua, L and Abwai, TT and Chand, D and Rama, V and Deo, V and Deo, KK and Tavuii, A and Valentine, W and Prasad, R and Seru, E and Naituku, L and Ratu, A and Hesketh, M and Kenny, N and Beebe, SC and Goundar, AA and McCaw, A and Buntine, M and Green, B and Frossard, T and Gilles, JRL and Joubert, DA and Wilson, G and Duong, LQ and Bouvier, JB and Stanford, D and Forder, C and Duyvestyn, JM and Pacidônio, EC and Flores, HA and Wittmeier, N and Retzki, K and Ryan, PA and Denton, JA and Smithyman, R and Tanamas, SK and Kyrylos, P and Dong, Y and Khalid, A and Hodgson, L and Anders, KL and O'Neill, SL}, title = {Successful introgression of wMel Wolbachia into Aedes aegypti populations in Fiji, Vanuatu and Kiribati.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {3}, pages = {e0012022}, doi = {10.1371/journal.pntd.0012022}, pmid = {38484041}, issn = {1935-2735}, abstract = {Pacific Island countries have experienced periodic dengue, chikungunya and Zika outbreaks for decades. The prevention and control of these mosquito-borne diseases rely heavily on control of Aedes aegypti mosquitoes, which in most settings are the primary vector. Introgression of the intracellular bacterium Wolbachia pipientis (wMel strain) into Ae. aegypti populations reduces their vector competence and consequently lowers dengue incidence in the human population. Here we describe successful area-wide deployments of wMel-infected Ae. aegypti in Suva, Lautoka, Nadi (Fiji), Port Vila (Vanuatu) and South Tarawa (Kiribati). With community support, weekly releases of wMel-infected Ae. aegypti mosquitoes for between 2 to 5 months resulted in wMel introgression in nearly all locations. Long term monitoring confirmed a high, self-sustaining prevalence of wMel infecting mosquitoes in almost all deployment areas. Measurement of public health outcomes were disrupted by the Covid19 pandemic but are expected to emerge in the coming years.}, } @article {pmid38483475, year = {2024}, author = {Maciel-de-Freitas, R and Sauer, FG and Kliemke, K and Garcia, GA and Pavan, MG and David, MR and Schmidt-Chanasit, J and Hoffmann, A and Lühken, R}, title = {Wolbachia strains wMel and wAlbB differentially affect Aedes aegypti traits related to fecundity.}, journal = {Microbiology spectrum}, volume = {}, number = {}, pages = {e0012824}, doi = {10.1128/spectrum.00128-24}, pmid = {38483475}, issn = {2165-0497}, abstract = {Two Wolbachia strains, wMel and wAlbB, have been transinfected into Aedes aegypti mosquitoes for population replacement with the aim of reducing dengue transmission. Epidemiological data from various endemic sites suggest a pronounced decrease in dengue transmission after implementing this strategy. In this study, we investigated the impact of the Wolbachia strains wMel and wAlbB on Ae. aegypti fitness in a common genetic background. We found that Ae. aegypti females infected with the wMel strain exhibited several significant differences compared with those infected with the wAlbB strain. Specifically, wMel-infected females laid significantly fewer eggs, ingested a lower amount of blood, had a reduced egg production rate, and exhibited a decreased Wolbachia density at a later age compared with mosquitoes infected with the wAlbB strain. Conversely, the wAlbB strain showed only mild negative effects when compared with Wolbachia-uninfected specimens. These differential effects on Ae. aegypti fitness following infection with either wMel or wAlbB may have important implications for the success of population replacement strategies in invading native Ae. aegypti populations in endemic settings. Further research is needed to better understand the underlying mechanisms responsible for these differences in fitness effects and their potential impact on the long-term efficacy of Wolbachia-based dengue control programs.IMPORTANCEThe transmission of arboviruses such as dengue, Zika, and chikungunya is on the rise globally. Among the most promising strategies to reduce arbovirus burden is the release of one out of two strains of Wolbachia-infected Aedes aegypti: wMel and wAlbB. One critical aspect of whether this approach will succeed involves the fitness cost of either Wolbachia strains on mosquito life history traits. For instance, we found that wMel-infected Ae. aegypti females laid significantly fewer eggs, ingested a lower amount of blood, had a reduced egg production rate, and exhibited a decreased Wolbachia density at a later age compared with mosquitoes infected with the wAlbB strain. Conversely, the wAlbB strain showed only mild negative effects when compared with Wolbachia-uninfected specimens. These differential effects on mosquito fitness following infection with either wMel or wAlbB may have important implications for the success of population replacement strategies in invading native Ae. aegypti populations.}, } @article {pmid38470882, year = {2024}, author = {Auguste, A and Ris, N and Belgaidi, Z and Kremmer, L and Mouton, L and Fauvergue, X}, title = {Insect population dynamics under Wolbachia-induced cytoplasmic incompatibility: Puzzle more than buzz in Drosophila suzukii.}, journal = {PloS one}, volume = {19}, number = {3}, pages = {e0300248}, doi = {10.1371/journal.pone.0300248}, pmid = {38470882}, issn = {1932-6203}, abstract = {In theory, the introduction of individuals infected with an incompatible strain of Wolbachia pipientis into a recipient host population should result in the symbiont invasion and reproductive failures caused by cytoplasmic incompatibility (CI). Modelling studies combining Wolbachia invasion and host population dynamics show that these two processes could interact to cause a transient population decline and, in some conditions, extinction. However, these effects could be sensitive to density dependence, with the Allee effect increasing the probability of extinction, and competition reducing the demographic impact of CI. We tested these predictions with laboratory experiments in the fruit fly Drosophila suzukii and the transinfected Wolbachia strain wTei. Surprisingly, the introduction of wTei into D. suzukii populations at carrying capacity did not result in the expected wTei invasion and transient population decline. In parallel, we found no Allee effect but strong negative density dependence. From these results, we propose that competition interacts in an antagonistic way with Wolbachia-induced cytoplasmic incompatibility on insect population dynamics. If future models and data support this hypothesis, pest management strategies using Wolbachia-induced CI should target populations with negligible competition but a potential Allee effect, for instance at the beginning of the reproductive season.}, } @article {pmid38469495, year = {2023}, author = {Field, EN and Smith, RC}, title = {Seasonality influences key physiological components contributing to Culex pipiens vector competence.}, journal = {Frontiers in insect science}, volume = {3}, number = {}, pages = {1144072}, pmid = {38469495}, issn = {2673-8600}, abstract = {Mosquitoes are the most important animal vector of disease on the planet, transmitting a variety of pathogens of both medical and veterinary importance. Mosquito-borne diseases display distinct seasonal patterns driven by both environmental and biological variables. However, an important, yet unexplored component of these patterns is the potential for seasonal influences on mosquito physiology that may ultimately influence vector competence. To address this question, we selected Culex pipiens, a primary vector of the West Nile virus (WNV) in the temperate United States, to examine the seasonal impacts on mosquito physiology by examining known immune and bacterial components implicated in mosquito arbovirus infection. Semi-field experiments were performed under spring, summer, and late-summer conditions, corresponding to historically low-, medium-, and high-intensity periods of WNV transmission, respectively. Through these experiments, we observed differences in the expression of immune genes and RNA interference (RNAi) pathway components, as well as changes in the distribution and abundance of Wolbachia in the mosquitoes across seasonal cohorts. Together, these findings support the conclusion that seasonal changes significantly influence mosquito physiology and components of the mosquito microbiome, suggesting that seasonality may impact mosquito susceptibility to pathogen infection, which could account for the temporal patterns in mosquito-borne disease transmission.}, } @article {pmid38468292, year = {2024}, author = {Ahouandjinou, MJ and Sovi, A and Sidick, A and Sewadé, W and Koukpo, CZ and Chitou, S and Towakinou, L and Adjottin, B and Hougbe, S and Tokponnon, F and Padonou, GG and Akogbéto, M and Messenger, LA and Ossè, RA}, title = {First report of natural infection of Anopheles gambiae s.s. and Anopheles coluzzii by Wolbachia and Microsporidia in Benin: a cross-sectional study.}, journal = {Malaria journal}, volume = {23}, number = {1}, pages = {72}, pmid = {38468292}, issn = {1475-2875}, abstract = {BACKGROUND: Recently, bacterial endosymbiont, including Wolbachia and Microsporidia were found to limit the infection of Anopheles mosquitoes with Plasmodium falciparum. This study aimed to investigate the natural presence of key transmission-blocking endosymbionts in Anopheles gambiae and Anopheles coluzzii in Southern Benin.

METHODS: The present study was conducted in seven communes (Cotonou, Porto-Novo, Aguégués, Ifangni, Pobè Athiémé, and Grand-Popo) of Southern Benin. Anopheles were collected using indoor/outdoor Human Landing Catches (HLCs) and Pyrethrum Spray Catches (PSCs). Following morphological identification, PCR was used to identify An. gambiae sensu lato (s.l.) to species level and to screen for the presence of both Wolbachia and Microsporidia. Plasmodium falciparum sporozoite infection was also assessed using ELISA.

RESULTS: Overall, species composition in An. gambiae s.l. was 53.7% An. coluzzii, while the remainder was An. gambiae sensu stricto (s.s.). Combined data of the two sampling techniques revealed a mean infection prevalence with Wolbachia of 5.1% (95% CI 0.90-18.6) and 1.3% (95% CI 0.07-7.8) in An. gambiae s.s. and An. coluzzii, respectively. The mean infection prevalence with Microsporidia was 41.0% (95% CI 25.9-57.8) for An. gambiae s.s. and 57.0% (95% CI 45.4-67.9) for An. coluzzii. Wolbachia was only observed in Ifangni, Pobè, and Cotonou, while Microsporidia was detected in all study communes. Aggregated data for HLCs and PSCs showed a sporozoite rate (SR) of 0.80% (95% CI 0.09-2.87) and 0.69% (95% CI 0.09-2.87) for An. gambiae and An. coluzzii, respectively, with a mean of 0.74% (95% CI 0.20-1.90). Of the four individual mosquitoes which harboured P. falciparum, none were also infected with Wolbachia and one contained Microsporidia.

CONCLUSIONS: The present study is the first report of natural infections of field-collected An. gambiae s.l. populations from Benin with Wolbachia and Microsporidia. Sustained efforts should be made to widen the spectrum of bacteria identified in mosquitoes, with the potential to develop endosymbiont-based control tools; such interventions could be the game-changer in the control of malaria and arboviral disease transmission.}, } @article {pmid38468766, year = {2022}, author = {Fernández, MB and Bleidorn, C and Calcaterra, LA}, title = {Wolbachia Infection in Native Populations of the Invasive Tawny Crazy Ant Nylanderia fulva.}, journal = {Frontiers in insect science}, volume = {2}, number = {}, pages = {905803}, pmid = {38468766}, issn = {2673-8600}, abstract = {Antagonistic interactions can affect population growth and dispersal of an invasive species. Wolbachia are intracellular endosymbiont bacteria that infect arthropod and nematode hosts and are able to manipulate reproduction, which in some cases leads to cocladogenesis. Moreover, the presence of the strictly maternally transferred Wolbachia in a population can indirectly induce selective sweeps on the hosts' mitochondria. Ants have a Wolbachia infection rate of about 34%, which makes phylogenetic studies using mitochondrial markers vulnerable of being confounded by the effect of the endosymbiont. Nylanderia fulva is an invasive ant native to South America, considered a pest in the United States. Its distribution and biology are poorly known in its native range, and the taxonomic identity of this and its closely related species, Nylanderia pubens, has only recently been understood with the aid of molecular phylogenies. Aiming at estimating robust phylogenetic relationships of N. fulva in its native range, we investigated the presence and pattern of Wolbachia infection in populations of N. fulva from Argentina, part of its native range, to account for its possible effect on the host population structure. Using the ftsZ gene, 30 nests of N. fulva and four from sympatric Nylanderia species were screened for the presence of Wolbachia. We sequenced the MLST genes, the highly variable gene wsp, as well as glyQ, a novel target gene for which new primers were designed. Phylogeny of the ants was estimated using mtDNA (COI). We found supergroup A Wolbachia strains infecting 73% of N. fulva nests and two nests of Nylanderia sp. 1. Wolbachia phylogenetic tree inferred with MLST genes is partially congruent with the host phylogeny topology, with the exception of a lineage of strains shared by ants from different N. fulva clades. Furthermore, by comparing with Wolbachia sequences infecting other ants, we found that the strains infecting different N. fulva clades are not monophyletic. Our findings suggest there are three recent independent horizontally transmitted Wolbachia infections in N. fulva, and we found no evidence of influence of Wolbachia in the host mtDNA based phylogeny.}, } @article {pmid38461924, year = {2024}, author = {Ateutchia-Ngouanet, S and Nanfack-Minkeu, F and Mavridis, K and Wanji, S and Demanou, M and Vontas, J and Djouaka, R}, title = {Monitoring Aedes populations for arboviruses, Wolbachia, insecticide resistance and its mechanisms in various agroecosystems in Benin.}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107178}, doi = {10.1016/j.actatropica.2024.107178}, pmid = {38461924}, issn = {1873-6254}, abstract = {Aedes mosquitoes are the main vectors of arboviruses in Benin. Cases of dengue have been reported in Benin with all four serotypes of the virus actively circulating in this region. Some agricultural settings are known to harbor Aedes vectors responsible for the transmission of arboviruses. The massive use of certain insecticides in agricultural settings has probably contributed to insecticide resistance in these vectors. In Benin, the susceptibility of arbovirus vectors to insecticides is poorly studied. In addition, the distribution of Wolbachia spp., which is used against some arboviruses is unknown. Moreover, there is limited information regarding the vectors responsible for the transmission of arboviruses in Benin. This present study monitored the species composition, arboviruses, and Wolbachia symbiont status, as well as the phenotypic and molecular insecticide resistance profile of Aedes populations from three agroecosystems in Benin. Aedes species identification was performed morphologically and confirmed using qPCR. (RT)-qPCR assay was applied for monitoring the presence of DENV, CHIKV, ZIKV, and WNV pathogens as well as for naturally occurring Wolbachia symbionts. Insecticide resistance was assessed phenotypically, by permethrin (0.75%) exposure of Adults (F0) using World Health Organization (WHO) bioassay protocols, and at the molecular level, using TaqMan (RT)-qPCR assays for assessing knock-down resistance (kdr) mutations (F1534C, V1016G/I, and S989P) and the expression levels of eight detoxification genes (P450s from the CYP9 and CYP6 families, carboxylesterases and glutathione-S-transferases). Aedes aegypti (Ae. aegypti) mosquitoes were the most abundant (93.9%) in the three agroecosystems studied, followed by Aedes albopictus (Ae. albopictus) mosquitoes (6.1%). No arboviruses were detected in the study's mosquito populations. Naturally occurring Wolbachia symbionts were present in 7 pools out of 15 pools tested. This could influence the effectiveness of vector control strategies based on exogenously introduced Wolbachia, all present in the three agroecosystems. Full susceptibility to permethrin was observed in all tested populations of Ae. albopictus. On the contrary, Ae. aegypti were found to be resistant in all three agroecosystem sites except for banana plantation sites, where full susceptibility was observed. Molecular analysis revealed that individual target site resistance kdr mutations F1534C and V1016G/I were detected in most Ae. aegypti populations. Additionally, double mutant (F1534C + V1016G/I) mosquitoes were found in some populations, and in one case, triple mutant (F1534C + V1016G/I + S989P) mosquitoes were detected. Metabolic resistance, as reflected by overexpression of three P450 genes (CYP6BB2, CYP9J26, and CYP9J32), was also detected in Ae. aegypti mosquitoes. Our study provides information that could be used to strategize future vector control strategies and highlights the importance of continuing vector surveillance. Future studies should assess the effect of piperonyl butoxide (PBO) on metabolic resistance and identify the different strains of Wolbachia spp., to choose the best vector control strategies in Benin.}, } @article {pmid38460359, year = {2024}, author = {Zurita, A and Trujillo, I and Cutillas, C}, title = {New records of pathogenic bacteria in different species of fleas collected from domestic and peridomestic animals in Spain. A potential zoonotic threat?.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {107}, number = {}, pages = {102153}, doi = {10.1016/j.cimid.2024.102153}, pmid = {38460359}, issn = {1878-1667}, abstract = {Climate change is causing many vectors of infectious diseases to expand their geographic distribution as well as the pathogens they transmit are also conditioned by temperature for their multiplication. Within this context, it is worth highlighting the significant role that fleas can play as vectors of important pathogenic bacteria. For this purpose, our efforts focused on detecting and identifying a total of 9 bacterial genera (Rickettsia sp.; Bartonella sp.; Yersinia sp.; Wolbachia sp., Mycobacterium sp., Leishmania sp., Borrelia sp., Francisella sp. and Coxiella sp.) within fleas isolated from domestic and peridomestic animals in the southwestern region of Spain (Andalusia). Over a 19-months period, we obtained flea samples from dogs, cats and hedgehogs. A total of 812 fleas was collected for this study. Five different species were morphologically identified, including C. felis, C. canis, S. cuniculi, P. irritans, and A. erinacei. Wolbachia sp. was detected in all five species identified in our study which a total prevalence of 86%. Within Rickettsia genus, two different species, R. felis and R. asembonensis were mainly identified in C. felis and A. erinacei, respectively. On the other hand, our results revealed a total of 131 fleas testing positive for the presence of Bartonella sp., representing a prevalence rate of 16% for this genus identifying two species B. henselae and B. clarridgeiae. Lastly, both Y. pestis and L. infantum were detected in DNA of P. irritans and C. felis, respectively isolated from dogs. With these data we update the list of bacterial zoonotic agents found in fleas in Spain, emphasizing the need to continue conducting future experimental studies to assess and confirm the potential vectorial role of certain synanthropic fleas.}, } @article {pmid38458883, year = {2024}, author = {Moore, CO and André, MR and Šlapeta, J and Breitschwerdt, EB}, title = {Vector biology of the cat flea Ctenocephalides felis.}, journal = {Trends in parasitology}, volume = {}, number = {}, pages = {}, doi = {10.1016/j.pt.2024.02.006}, pmid = {38458883}, issn = {1471-5007}, abstract = {Ctenocephalides felis, the cat flea, is among the most prevalent and widely dispersed vectors worldwide. Unfortunately, research on C. felis and associated pathogens (Bartonella and Rickettsia spp.) lags behind that of other vectors and vector-borne pathogens. Therefore, we aimed to review fundamental aspects of C. felis as a vector (behavior, epidemiology, phylogenetics, immunology, and microbiome composition) with an emphasis on key techniques and research avenues employed in other vector species. Future laboratory C. felis experimental infections with Bartonella, Rickettsia, and Wolbachia species/strains should examine the vector-pathogen interface utilizing contemporary visualization, transcriptomic, and gene-editing techniques. Further environmental sampling will inform the range and prevalence of C. felis and associated pathogens, improving the accuracy of vector and pathogen modeling to improve infection/infestation risk assessment and diagnostic recommendations.}, } @article {pmid38454541, year = {2024}, author = {Carlassara, M and Khorramnejad, A and Oker, H and Bahrami, R and Lozada-Chávez, AN and Mancini, MV and Quaranta, S and Body, MJA and Lahondère, C and Bonizzoni, M}, title = {Population-specific responses to developmental temperature in the arboviral vector Aedes albopictus: Implications for climate change.}, journal = {Global change biology}, volume = {30}, number = {3}, pages = {e17226}, doi = {10.1111/gcb.17226}, pmid = {38454541}, issn = {1365-2486}, support = {//2022J45MLL/ ; PE00000007//NextGeneration EU-MUR PNRR Extended Partnership/ ; }, abstract = {The increase of environmental temperature due to current global warming is not only favouring the expansion of the distribution range of many insect species, but it is also changing their phenology. Insect phenology is tightly linked to developmental timing, which is regulated by environmental temperatures. However, the degree to which the effects of developmental temperatures extend across developmental stages and their inter-stage relationships have not been thoroughly quantified in mosquitoes. Here, we used the mosquito Aedes albopictus, which is an aggressive invasive species and an arboviral vector, to study how developmental temperature influences fitness across developmental stages, thermal traits, energy reserves, transcriptome and Wolbachia prevalence in laboratory-reared populations originally collected from either temperate or tropical regions. We show that hatchability, larval and pupal viability and developmental speed are strongly influenced by temperature, and these effects extend to wing length, body mass, longevity and content of water, protein and lipids in adults in a population-specific manner. On the contrary, neither adult thermal preference nor heat resistance significantly change with temperature. Wolbachia density was generally lower in adult mosquitoes reared at 18°C than at other tested temperatures, and transcriptome analysis showed enrichment for functions linked to stress responses (i.e. cuticle proteins and chitin, cytochrome p450 and heat shock proteins) in mosquitoes reared at both 18 and 32°C. Our data showed an overall reduced vector fitness performance when mosquitoes were reared at 32°C, and the absence of isomorphy in the relationship between developmental stages and temperature in the laboratory population deriving from larvae collected in northern Italy. Altogether, these results have important implications for reliable model projections of the invasion potentials of Ae. albopictus and its epidemiological impact.}, } @article {pmid38452081, year = {2024}, author = {Kaur, R and McGarry, A and Shropshire, JD and Leigh, BA and Bordenstein, SR}, title = {Prophage proteins alter long noncoding RNA and DNA of developing sperm to induce a paternal-effect lethality.}, journal = {Science (New York, N.Y.)}, volume = {383}, number = {6687}, pages = {1111-1117}, doi = {10.1126/science.adk9469}, pmid = {38452081}, issn = {1095-9203}, abstract = {The extent to which prophage proteins interact with eukaryotic macromolecules is largely unknown. In this work, we show that cytoplasmic incompatibility factor A (CifA) and B (CifB) proteins, encoded by prophage WO of the endosymbiont Wolbachia, alter long noncoding RNA (lncRNA) and DNA during Drosophila sperm development to establish a paternal-effect embryonic lethality known as cytoplasmic incompatibility (CI). CifA is a ribonuclease (RNase) that depletes a spermatocyte lncRNA important for the histone-to-protamine transition of spermiogenesis. Both CifA and CifB are deoxyribonucleases (DNases) that elevate DNA damage in late spermiogenesis. lncRNA knockdown enhances CI, and mutagenesis links lncRNA depletion and subsequent sperm chromatin integrity changes to embryonic DNA damage and CI. Hence, prophage proteins interact with eukaryotic macromolecules during gametogenesis to create a symbiosis that is fundamental to insect evolution and vector control.}, } @article {pmid38450861, year = {2024}, author = {Urakova, N and Joseph, RE and Huntsinger, A and Macias, VM and Jones, MJ and Sigle, LT and Li, M and Akbari, OS and Xi, Z and Lymperopoulos, K and Sayre, RT and McGraw, EA and Rasgon, JL}, title = {Alpha-mannosidase-2 modulates arbovirus infection in a pathogen- and Wolbachia-specific manner in Aedes aegypti mosquitoes.}, journal = {Insect molecular biology}, volume = {}, number = {}, pages = {}, doi = {10.1111/imb.12904}, pmid = {38450861}, issn = {1365-2583}, support = {R01AI116636/NH/NIH HHS/United States ; R01AI150251/NH/NIH HHS/United States ; R01AI143758/NH/NIH HHS/United States ; R01AI151004/NH/NIH HHS/United States ; DP2AI152071/NH/NIH HHS/United States ; R21AI156078/NH/NIH HHS/United States ; }, abstract = {Multiple Wolbachia strains can block pathogen infection, replication and/or transmission in Aedes aegypti mosquitoes under both laboratory and field conditions. However, Wolbachia effects on pathogens can be highly variable across systems and the factors governing this variability are not well understood. It is increasingly clear that the mosquito host is not a passive player in which Wolbachia governs pathogen transmission phenotypes; rather, the genetics of the host can significantly modulate Wolbachia-mediated pathogen blocking. Specifically, previous work linked variation in Wolbachia pathogen blocking to polymorphisms in the mosquito alpha-mannosidase-2 (αMan2) gene. Here we use CRISPR-Cas9 mutagenesis to functionally test this association. We developed αMan2 knockouts and examined effects on both Wolbachia and virus levels, using dengue virus (DENV; Flaviviridae) and Mayaro virus (MAYV; Togaviridae). Wolbachia titres were significantly elevated in αMan2 knockout (KO) mosquitoes, but there were complex interactions with virus infection and replication. In Wolbachia-uninfected mosquitoes, the αMan2 KO mutation was associated with decreased DENV titres, but in a Wolbachia-infected background, the αMan2 KO mutation significantly increased virus titres. In contrast, the αMan2 KO mutation significantly increased MAYV replication in Wolbachia-uninfected mosquitoes and did not affect Wolbachia-mediated virus blocking. These results demonstrate that αMan2 modulates arbovirus infection in A. aegypti mosquitoes in a pathogen- and Wolbachia-specific manner, and that Wolbachia-mediated pathogen blocking is a complex phenotype dependent on the mosquito host genotype and the pathogen. These results have a significant impact for the design and use of Wolbachia-based strategies to control vector-borne pathogens.}, } @article {pmid38450576, year = {2024}, author = {Mancini, MV and Murdochy, SM and Bilgo, E and Ant, TH and Gingell, D and Gnambani, EJ and Failloux, AB and Diabate, A and Sinkins, SP}, title = {Wolbachia strain wAlbB shows favourable characteristics for dengue control use in Aedes aegypti from Burkina Faso.}, journal = {Environmental microbiology}, volume = {26}, number = {3}, pages = {e16588}, doi = {10.1111/1462-2920.16588}, pmid = {38450576}, issn = {1462-2920}, support = {AV/PP0025/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; 226166/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Dengue represents an increasing public health burden worldwide. In Africa, underreporting and misdiagnosis often mask its true epidemiology, and dengue is likely to be both more widespread than reported data suggest and increasing in incidence and distribution. Wolbachia-based dengue control is underway in Asia and the Americas but has not to date been deployed in Africa. Due to the genetic heterogeneity of African Aedes aegypti populations and the complexity of the host-symbiont interactions, characterization of key parameters of Wolbachia-carrying mosquitoes is paramount for determining the potential of the system as a control tool for dengue in Africa. The wAlbB Wolbachia strain was stably introduced into an African Ae. aegypti population by introgression, and showed high intracellular density in whole bodies and different mosquito tissues; high intracellular density was also maintained following larval rearing at high temperatures. No effect on the adult lifespan induced by Wolbachia presence was detected. Moreover, the ability of this strain to strongly inhibit DENV-2 dissemination and transmission in the host was also demonstrated in the African background. Our findings suggest the potential of harnessing Wolbachia for dengue control for African populations of Ae. aegypti.}, } @article {pmid38446807, year = {2024}, author = {Pérez-Guerra, CL and Rosado-Santiago, C and Ramos, SA and Marrero-Santos, KM and González-Zeno, G and Partridge, SK and Rivera-Amill, V and Paz-Bailey, G and Sánchez-González, L and Hayden, MH}, title = {Acceptability of emergent Aedes aegypti vector control methods in Ponce, Puerto Rico: A qualitative assessment.}, journal = {PLOS global public health}, volume = {4}, number = {3}, pages = {e0002744}, doi = {10.1371/journal.pgph.0002744}, pmid = {38446807}, issn = {2767-3375}, abstract = {Aedes aegypti control has been fraught with challenges in Puerto Rico. The government has implemented commonly used vector control methods, but arboviral epidemics still occur. It is necessary to explore new Ae. aegypti control methods. This study aimed to understand the perceptions of community members in Ponce, Puerto Rico about emergent and traditional Ae. aegypti vector control methods and determine their acceptability and support for these methods. We identified the type of information needed to increase support for emergent vector control methods, and the preferred strategies to disseminate this information. Four group discussions were conducted with a total of 32 participants representing eight of the 14 clusters participating in the Communities Organized for the Prevention of Arboviruses (COPA), a project designed to mobilize communities in Ponce, Puerto Rico to prevent diseases transmitted by mosquitoes. Group discussions began with an overview of different methods used for controlling Ae. aegypti mosquitoes. These overviews facilitated participant understanding of the mosquito control methods presented. Use of source reduction, autocidal gravid ovitraps (AGO), and manual application of larvicide for arboviral mosquito control received support from almost all participants. Vector control methods that use more familiar techniques in Puerto Rico such as truck-mounted larvicide spraying (TMLS) and insecticide residual spraying received support from most participants. More than half of participants supported the use of emergent mosquito control methods including Wolbachia suppression, Wolbachia replacement, or genetically modified mosquitoes (GMM). Participants preferred to receive vector control information through house-to-house visits with the distribution of written materials, followed by dissemination of information through traditional (i.e., radio, television) and social media. The detailed information resulting from this study was used to develop messages for a communications campaign to garner future community support. Community acceptance and support are critical for the success of vector control programs using emergent mosquito control methods.}, } @article {pmid38437189, year = {2024}, author = {Dye, D and Cain, JW}, title = {Efficacy of Wolbachia-based mosquito control: Predictions of a spatially discrete mathematical model.}, journal = {PloS one}, volume = {19}, number = {3}, pages = {e0297964}, doi = {10.1371/journal.pone.0297964}, pmid = {38437189}, issn = {1932-6203}, abstract = {Wolbachia is an endosymbiont bacterium present in many insect species. When Wolbachia-carrying male Aedes aegypti mosquitoes mate with non-carrier females, their embryos are not viable due to cytoplasmic incompatibility. This phenomenon has been exploited successfully for the purpose of controlling mosquito populations and the spread of mosquito-borne illnesses: Wolbachia carriers are bred and released into the environment. Because Wolbachia is not harmful to humans, this method of mosquito control is regarded as a safer alternative to pesticide spraying. In this article, we introduce a mathematical framework for exploring (i) whether a one-time release of Wolbachia carriers can elicit a sustained presence of carriers near the release site, and (ii) the extent to which spatial propagation of carriers may allow them to establish fixation in other territories. While some prior studies have formulated mosquito dispersal models using advection-reaction-diffusion PDEs, the predictive power of such models requires careful ecological mapping: advection and diffusion coefficients exhibit significant spatial dependence due to heterogeneity of resources and topography. Here, we adopt a courser-grained view, regarding the environment as a network of discrete, diffusively-coupled "habitats"-distinct zones of high mosquito density such as stagnant ponds. We extend two previously published single-habitat mosquito models to multiple habitats, and calculate rates of migration between pairs of habitats using dispersal kernels. Our primary results are quantitative estimates regarding how the success of carrier fixation in one or more habitats is determined by: the number of carriers released, sizes of habitats, distances between habitats, and the rate of migration between habitats. Besides yielding sensible and potentially useful predictions regarding the success of Wolbachia-based control, our framework applies to other approaches (e.g., gene drives) and contexts beyond the realm of insect pest control.}, } @article {pmid38431055, year = {2024}, author = {Walt, HK and King, JG and Sheele, JM and Meyer, F and Pietri, JE and Hoffmann, FG}, title = {Do bed bugs transmit human viruses, or do humans spread bed bugs and their viruses? A worldwide survey of the bed bug RNA virosphere.}, journal = {Virus research}, volume = {}, number = {}, pages = {199349}, doi = {10.1016/j.virusres.2024.199349}, pmid = {38431055}, issn = {1872-7492}, abstract = {BED BUGS (HEMIPTERA: : Cimicidae) are a globally distributed hematophagous pest that routinely feed on humans. Unlike many blood-sucking arthropods, they have never been linked to pathogen transmission in a natural setting, and despite increasing interest in their role as disease vectors, little is known about the viruses that bed bugs naturally harbor. Here, we present a global-scale survey of the bed bug RNA virosphere. We sequenced the metatranscriptomes of 22 individual bed bugs (Cimex lectularius and Cimex hemipterus) from 8 locations around the world. We detected sequences from two known bed bug viruses (Shuangao bedbug virus 1 and Shuangao bedbug virus 2) which extends their geographical range. We identified three novel bed bug virus sequences from a tenui-like virus (Bunyavirales), a toti-like virus (Ghabrivirales), and a luteo-like virus (Tolivirales). Interestingly, some of the bed bug viruses branch near to insect-transmitted plant-infecting viruses, opening questions regarding the evolution of plant virus infection. When we analyzed the viral sequences by their host's collection location, we found unexpected patterns of geographical diversity that may reflect humans' role in bed bug dispersal. Additionally, we investigated the effect that Wolbachia, the primary bed bug endosymbiont, may have on viral abundance and found that Wolbachia infection neither promotes nor inhibits viral infection. Finally, our results provide no evidence that bed bugs transmit any known human pathogenic viruses.}, } @article {pmid38427588, year = {2024}, author = {Aldridge, RL and Gibson, S and Linthicum, KJ}, title = {Aedes aegypti Controls AE. Aegypti: SIT and IIT-An Overview.}, journal = {Journal of the American Mosquito Control Association}, volume = {40}, number = {1}, pages = {32-49}, doi = {10.2987/23-7154}, pmid = {38427588}, issn = {1943-6270}, abstract = {The sterile insect technique (SIT) and the incompatible insect technique (IIT) are emerging and potentially revolutionary tools for controlling Aedes aegypti (L.), a prominent worldwide mosquito vector threat to humans that is notoriously difficult to reduce or eliminate in intervention areas using traditional integrated vector management (IVM) approaches. Here we provide an overview of the discovery, development, and application of SIT and IIT to Ae. aegypti control, and innovations and advances in technology, including transgenics, that could elevate these techniques to a worldwide sustainable solution to Ae. aegypti when combined with other IVM practices.}, } @article {pmid38426058, year = {2024}, author = {Cantin, LJ and Dunning Hotopp, JC and Foster, JM}, title = {Improved metagenome assemblies through selective enrichment of bacterial genomic DNA from eukaryotic host genomic DNA using ATAC-seq.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1352378}, doi = {10.3389/fmicb.2024.1352378}, pmid = {38426058}, issn = {1664-302X}, abstract = {Genomics can be used to study the complex relationships between hosts and their microbiota. Many bacteria cannot be cultured in the laboratory, making it difficult to obtain adequate amounts of bacterial DNA and to limit host DNA contamination for the construction of metagenome-assembled genomes (MAGs). For example, Wolbachia is a genus of exclusively obligate intracellular bacteria that live in a wide range of arthropods and some nematodes. While Wolbachia endosymbionts are frequently described as facultative reproductive parasites in arthropods, the bacteria are obligate mutualistic endosymbionts of filarial worms. Here, we achieve 50-fold enrichment of bacterial sequences using ATAC-seq (Assay for Transposase-Accessible Chromatin using sequencing) with Brugia malayi nematodes, containing Wolbachia (wBm). ATAC-seq uses the Tn5 transposase to cut and attach Illumina sequencing adapters to accessible DNA lacking histones, typically thought to be open chromatin. Bacterial and mitochondrial DNA in the lysates are also cut preferentially since they lack histones, leading to the enrichment of these sequences. The benefits of this include minimal tissue input (<1 mg of tissue), a quick protocol (<4 h), low sequencing costs, less bias, correct assembly of lateral gene transfers and no prior sequence knowledge required. We assembled the wBm genome with as few as 1 million Illumina short paired-end reads with >97% coverage of the published genome, compared to only 12% coverage with the standard gDNA libraries. We found significant bacterial sequence enrichment that facilitated genome assembly in previously published ATAC-seq data sets from human cells infected with Mycobacterium tuberculosis and C. elegans contaminated with their food source, the OP50 strain of E. coli. These results demonstrate the feasibility and benefits of using ATAC-seq to easily obtain bacterial genomes to aid in symbiosis, infectious disease, and microbiome research.}, } @article {pmid38412904, year = {2024}, author = {Liu, K and Fang, S and Li, Q and Lou, Y}, title = {Effectiveness evaluation of mosquito suppression strategies on dengue transmission under changing temperature and precipitation.}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107159}, doi = {10.1016/j.actatropica.2024.107159}, pmid = {38412904}, issn = {1873-6254}, abstract = {Widespread resurgence of dengue outbreaks has seriously threatened the global health. Due to lack of treatments and vaccines, one key strategy in dengue control is to reduce the vector population size. As an environment-friendly mosquito control approach, releasing male mosquitoes transinfected with specific Wolbachia strain into the field to suppress the wild mosquito population size has become wildly accepted. The current study evaluates the effectiveness of this suppression strategy on dengue control under changing temperature and precipitation profiles. We formulate a mathematical model which includes larval intra-specific competition, the maturation period for mosquitoes, the extrinsic incubation period (EIP) and intrinsic incubation period (IIP). The persistence of mosquitoes and disease is discussed in terms of two basic reproduction numbers (RM and R0) and the release ratio pw. Further numerical simulations are carried out to not only validate theoretical results, but also provide interesting quantitative observations. Sensitivity analysis on the reproduction numbers, peak size, peak time and the final epidemic size is performed with respect to model parameters, which highlights effective control measures against dengue transmission. Moreover, by assuming temperature and precipitation dependent mosquito-related parameters, the model can be used to project the effectiveness of releasing Wolbachia-carrying males under climatic variations. It is shown that the effectiveness of various control strategies is highly dependent on the changing temperature and precipitation profiles. In particular, the model projects that it is most challenging to control the disease at the favorable temperature (around 27∼30[∘]C) and precipitation (5∼8mm/day) range, during which the basic reproduction number R0 is very high and more Wolbachia-infected males should be released.}, } @article {pmid38404602, year = {2024}, author = {Yan, J and Green, K and Noel, K and Kim, CH and Stone, CM}, title = {Effects of seasonality and developed land cover on Culex mosquito abundance and microbiome diversity.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1332970}, pmid = {38404602}, issn = {1664-302X}, abstract = {The vectorial capacity of mosquitoes, which influences the dynamics of vector-borne disease transmission, is intricately linked to mosquito abundance and the composition and diversity of their associated microbiomes. However, the influence of environmental factors on mosquito populations and microbiome diversity remains underexplored. Here we examined the effects of seasonality and developed land cover on Culex mosquito abundance and bacterial diversity. Biweekly field sampling of female Culex mosquitoes was conducted using gravid and CDC light traps, spanning summer to autumn across varying developed land cover levels in two urban areas in Central Illinois. Mosquito abundance was assessed by the number of mosquitoes captured per trap night and compared across seasons and developed levels. The mean mosquito abundance for gravid and light traps was 12.96 ± 2.15 and 7.67 ± 1.44, respectively. Notably, higher levels of developed land cover exhibited higher Culex abundance than the low level for light traps, but no significant difference was found between summer and early autumn. In gravid traps, no significant differences were detected across seasons or developed levels. Microbial analysis of the mosquito microbiome revealed that Proteobacteria and Wolbachia, with a mean relative abundance of 80.77 and 52.66% respectively, were identified as the most dominant bacterial phylum and genus. Their relative abundance remained consistent across seasons and developed land cover levels, with negligible variations. Alpha diversity, as measured by observed species, Chao1, Shannon, and Simpson, showed slightly higher values in early-autumn compared to late-summer. A notable pattern of bacterial diversity, as indicated by all four diversity indexes, is evident across varying levels of land development. Significantly, high or intermediate developed levels consistently showed reduced alpha diversity when compared to the lower level. This underscores the pronounced impact of anthropogenic ecological disturbances in shaping mosquito microbiomes. Beta diversity analysis revealed no significant dissimilarities in bacterial community composition across seasons and developed levels, although some separation was noted among different levels of developed land cover. These findings highlight the significant role of environmental factors in shaping mosquito abundance and their associated microbiomes, with potential implications for the vectorial capacity in the transmission of vector-borne diseases.}, } @article {pmid38399990, year = {2024}, author = {Corrêa-Antônio, J and David, MR and Couto-Lima, D and Garcia, GA and Keirsebelik, MSG and Maciel-de-Freitas, R and Pavan, MG}, title = {DENV-1 Titer Impacts Viral Blocking in wMel Aedes aegypti with Brazilian Genetic Background.}, journal = {Viruses}, volume = {16}, number = {2}, pages = {}, doi = {10.3390/v16020214}, pmid = {38399990}, issn = {1999-4915}, support = {E26/2001.844/2017//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; E26/210.335/2022//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; E26/201.370/2022//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; E26/211.159/2019//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; finance code 001//Coordenação de Aperfeicoamento de Pessoal de Nível Superior/ ; }, abstract = {Several countries have been using Wolbachia deployments to replace highly competent native Aedes aegypti populations with Wolbachia-carrying mosquitoes with lower susceptibility to arboviruses such as dengue, Zika, and chikungunya. In Rio de Janeiro, Wolbachia deployments started in 2015 and still present a moderate introgression with a modest reduction in dengue cases in humans (38%). Here, we evaluated the vector competence of wild-type and wMel-infected Ae. aegypti with a Brazilian genetic background to investigate whether virus leakage could contribute to the observed outcomes in Brazil. We collected the specimens in three areas of Rio de Janeiro with distinct frequencies of mosquitoes with wMel strain and two areas with wild Ae. aegypti. The mosquitoes were orally exposed to two titers of DENV-1 and the saliva of DENV-1-infected Ae. aegypti was microinjected into wMel-free mosquitoes to check their infectivity. When infected with the high DENV-1 titer, the presence of wMel did not avoid viral infection in mosquitoes' bodies and saliva but DENV-1-infected wMel mosquitoes produced lower viral loads than wMel-free mosquitoes. On the other hand, wMel mosquitoes infected with the low DENV-1 titer were less susceptible to virus infection than wMel-free mosquitoes, although once infected, wMel and wMel-free mosquitoes exhibited similar viral loads in the body and the saliva. Our results showed viral leakage in 60% of the saliva of wMel mosquitoes with Brazilian background; thus, sustained surveillance is imperative to monitor the presence of other circulating DENV-1 strains capable of overcoming the Wolbachia blocking phenotype, enabling timely implementation of action plans.}, } @article {pmid38399310, year = {2024}, author = {Voronin, D and Tricoche, N and Peguero, R and Kaminska, AM and Ghedin, E and Sakanari, JA and Lustigman, S}, title = {Repurposed Drugs That Activate Autophagy in Filarial Worms Act as Effective Macrofilaricides.}, journal = {Pharmaceutics}, volume = {16}, number = {2}, pages = {}, doi = {10.3390/pharmaceutics16020256}, pmid = {38399310}, issn = {1999-4923}, support = {R21AI153649//National Institute of Allergy and Infectious Diseases/ ; ZIA-AI001322//Division of Intramural Research of the NIAID/NIH/ ; }, abstract = {Onchocerciasis and lymphatic filariasis are two neglected tropical diseases caused by filarial nematodes that utilize insect vectors for transmission to their human hosts. Current control strategies are based on annual or biannual mass drug administration (MDA) of the drugs Ivermectin or Ivermectin plus Albendazole, respectively. These drug regimens kill the first-stage larvae of filarial worms (i.e., microfilariae) and interrupt the transmission of infections. MDA programs for these microfilaricidal drugs must be given over the lifetime of the filarial adult worms, which can reach 15 years in the case of Onchocerca volvulus. This is problematic because of suboptimal responses to ivermectin in various endemic regions and inefficient reduction of transmission even after decades of MDA. There is an urgent need for the development of novel alternative treatments to support the 2030 elimination goals of onchocerciasis and lymphatic filariasis. One successful approach has been to target Wolbachia, obligatory endosymbiotic bacteria on which filarial worms are dependent for their survival and reproduction within the human host. A 4-6-week antibiotic therapy with doxycycline, for example, resulted in the loss of Wolbachia that subsequently led to extensive apoptosis of somatic cells, germline, embryos, and microfilariae, as well as inhibition of fourth-stage larval development. However, this long-course regimen has limited use in MDA programs. As an alternative approach to the use of bacteriostatic antibiotics, in this study, we focused on autophagy-inducing compounds, which we hypothesized could disturb various pathways involved in the interdependency between Wolbachia and filarial worms. We demonstrated that several such compounds, including Niclosamide, an FDA-approved drug, Niclosamide ethanolamine (NEN), and Rottlerin, a natural product derived from Kamala trees, significantly reduced the levels of Wolbachia in vitro. Moreover, when these compounds were used in vivo to treat Brugia pahangi-infected gerbils, Niclosamide and NEN significantly decreased adult worm survival, reduced the release of microfilariae, and decreased embryonic development depending on the regimen and dose used. All three drugs given orally significantly reduced Wolbachia loads and induced an increase in levels of lysosome-associated membrane protein in worms from treated animals, suggesting that Niclosamide, NEN, and Rottlerin were effective in causing drug-induced autophagy in these filarial worms. These repurposed drugs provide a new avenue for the clearance of adult worms in filarial infections.}, } @article {pmid38394822, year = {2024}, author = {Zhang, X and Ferree, PM}, title = {PSRs: Selfish chromosomes that manipulate reproductive development.}, journal = {Seminars in cell & developmental biology}, volume = {159-160}, number = {}, pages = {66-73}, doi = {10.1016/j.semcdb.2024.01.008}, pmid = {38394822}, issn = {1096-3634}, abstract = {B chromosomes are intriguing "selfish" genetic elements, many of which exhibit higher-than-Mendelian transmission. This perspective highlights a group of B chromosomes known as Paternal Sex Ratio chromosomes (PSRs), which are found in several insects with haplo-diploid reproduction. PSRs harshly alter the organism's reproduction to facilitate their own inheritance. A manifestation of this effect is the conversion of female destined individuals into males. Key to this conversion is the mysterious ability of PSRs to cause elimination of the sperm-inherited half of the genome during zygote formation. Here we discuss how PSRs were discovered, what is known about how they alter paternal chromatin dynamics to cause sex conversion, and how PSR-induced genome elimination is different from other forms of programmed genome elimination in different insects. PSRs also stand out because their DNA sequence compositions differ in remarkable ways from their insect's essential chromosomes, a characteristic suggestive of interspecies origins. Broadly, we also highlight poorly understood aspects of PSR dynamics that need to be investigated.}, } @article {pmid38392845, year = {2024}, author = {Lamichhane, B and Brockway, C and Evasco, K and Nicholson, J and Neville, PJ and Levy, A and Smith, D and Imrie, A}, title = {Metatranscriptomic Sequencing of Medically Important Mosquitoes Reveals Extensive Diversity of RNA Viruses and Other Microbial Communities in Western Australia.}, journal = {Pathogens (Basel, Switzerland)}, volume = {13}, number = {2}, pages = {}, doi = {10.3390/pathogens13020107}, pmid = {38392845}, issn = {2076-0817}, abstract = {Mosquitoes harbor a wide diversity of microorganisms, including viruses that are human pathogens, or that are insect specific. We used metatranscriptomics, an unbiased high-throughput molecular approach, to describe the composition of viral and other microbial communities in six medically important mosquito species from across Western Australia: Aedes vigilax, Culex annulirostris, Cx. australicus, Cx. globocoxitus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. We identified 42 viral species, including 13 novel viruses, from 19 families. Culex mosquitoes exhibited a significantly higher diversity of viruses than Aedes mosquitoes, and no virus was shared between the two genera. Comparison of mosquito populations revealed a heterogenous distribution of viruses between geographical regions and between closely related species, suggesting that geography and host species may play a role in shaping virome composition. We also detected bacterial and parasitic microorganisms, among which Wolbachia bacteria were detected in three members of the Cx. pipiens complex, Cx. australicus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. In summary, our unbiased metatranscriptomics approach provides important insights into viral and other microbial diversity in Western Australian mosquitoes that vector medically important viruses.}, } @article {pmid38392551, year = {2024}, author = {Zhu, X and Li, J and He, A and Gurr, GM and You, M and You, S}, title = {Developmental Shifts in the Microbiome of a Cosmopolitan Pest: Unraveling the Role of Wolbachia and Dominant Bacteria.}, journal = {Insects}, volume = {15}, number = {2}, pages = {}, doi = {10.3390/insects15020132}, pmid = {38392551}, issn = {2075-4450}, support = {2022J06013//The Fujian Natural Science Fund for Distinguished Young Scholars/ ; 2022L3087//Central Guided Local Science and Technology Development Project/ ; }, abstract = {Wolbachia bacteria (phylum Proteobacteria) are ubiquitous intracellular parasites of diverse invertebrates. In insects, coevolution has forged mutualistic associations with Wolbachia species, influencing reproduction, immunity, development, pathogen resistance, and overall fitness. However, the impact of Wolbachia on other microbial associates within the insect microbiome, which are crucial for host fitness, remains less explored. The diamondback moth (Plutella xylostella), a major pest of cruciferous vegetables worldwide, harbors the dominant Wolbachia strain plutWB1, known to distort its sex ratio. This study investigated the bacterial community diversity and dynamics across different developmental life stages and Wolbachia infection states in P. xylostella using high-throughput 16S rDNA amplicon sequencing. Proteobacteria and Firmicutes dominated the P. xylostella microbiome regardless of life stage or Wolbachia infection. However, the relative abundance of dominant genera, including an unclassified genus of Enterobacteriaceae, Wolbachia, Carnobacterium, and Delftia tsuruhatensis, displayed significant stage-specific variations. While significant differences in bacterial diversity and composition were observed across life stages, Wolbachia infection had no substantial impact on overall diversity. Nonetheless, relative abundances of specific genera differed between infection states. Notably, Wolbachia exhibited a stable, high relative abundance across all stages and negatively correlated with an unclassified genus of Enterobacteriaceae, Delftia tsuruhatensis, and Carnobacterium. Our findings provide a foundational understanding of the complex interplay between the host, Wolbachia, and the associated microbiome in P. xylostella, paving the way for a deeper understanding of their complex interactions and potential implications for pest control strategies.}, } @article {pmid38392506, year = {2024}, author = {Budrys, E and Orlovskytė, S and Budrienė, A}, title = {Ecological Speciation without Morphological Differentiation? A New Cryptic Species of Diodontus Curtis (Hymenoptera, Pemphredonidae) from the Centre of Europe.}, journal = {Insects}, volume = {15}, number = {2}, pages = {}, doi = {10.3390/insects15020086}, pmid = {38392506}, issn = {2075-4450}, support = {Contract No S-MIP-20-23//Lietuvos Mokslo Taryba/ ; }, abstract = {Upon exploring the mitotype diversity of the aphid-hunting wasp, Diodontus tristis, we revealed specimens with highly divergent mitotypes from two localities in Lithuania and nesting in clayey substrate, while the specimens with typical mitotypes were found nesting in sandy sites. The comparison of inter- and intra-specific distances and application of delimitation algorithms supported the species status of the clay-nesting populations. Using a set of DNA markers that included complete or partial sequences of six mitochondrial genes, three markers of ribosomal operon, two homeobox genes, and four other nuclear genes, we clarified the phylogenetic relationships of the new cryptic species. The endosymbiotic bacteria infestation was checked, considering the option that the divergent populations may represent clades isolated by Wolbachia infection; however, it did not demonstrate any specificity. We found only subtle morphological differences in the new clay-nesting species, D. argillicola sp. nov.; the discriminant analysis of morphometric measurements did not reliably segregate it as well. Thus, we provide the molecular characters of the cryptic species, which allow confident identification, its phylogenetic position within the genus, and an updated identification key for the D. tristis species group.}, } @article {pmid38392339, year = {2024}, author = {Reese, C and Graber, LC and Ramalho, MO and Moreau, CS}, title = {The Diversity of Wolbachia across the Turtle Ants (Formicidae: Cephalotes spp.).}, journal = {Biology}, volume = {13}, number = {2}, pages = {}, doi = {10.3390/biology13020121}, pmid = {38392339}, issn = {2079-7737}, support = {DGE-1650441; NSF DEB 1900357//National Science Foundation/ ; }, abstract = {Wolbachia is a widespread and well-known bacterium that can induce a wide range of changes within its host. Ants specifically harbor a great deal of Wolbachia diversity and are useful systems to study endosymbiosis. The turtle ants (Cephalotes) are a widespread group of tropical ants that rely on gut microbes to support their herbivorous diet for their survival, yet little is known of the extent of this diversity. Therefore, studying their endosymbionts and categorizing the diversity of bacteria within Cephalotes hosts could help to delimit species and identify new strains and can help lead to a further understanding of how the microbiome leads to survival and speciation in the wild. In our study, 116 individual samples were initially tested for positive infection with the wsp gene. Of the initial 116 samples, 9 samples were infected with only one strain of Wolbachia, and 7 were able to be used successfully for multilocus sequence typing (MLST). We used the new MLST data to infer a phylogeny with other Formicidae samples from the MLST online database to identify new Wolbachia strains and related genes, of which only one came back as an exact match. The 18 Wolbachia-positive samples ranged across 15 different species and 7 different countries, which we further test for species identity and geographic correlation. This study is the first comprehensive look into the diversity of Wolbachia in the turtle ants, providing insight into how endosymbionts are oriented in widespread species and providing a strong foundation for further research in host-microbe interactions.}, } @article {pmid38388801, year = {2024}, author = {Ye, QT and Gong, X and Liu, HH and Wu, BX and Peng, CW and Hong, XY and Bing, XL}, title = {The symbiont Wolbachia alleviates pesticide susceptibility in the two-spotted spider mite Tetranychus urticae through enhanced host detoxification pathways.}, journal = {Insect science}, volume = {}, number = {}, pages = {}, doi = {10.1111/1744-7917.13341}, pmid = {38388801}, issn = {1744-7917}, support = {32020103011//National Natural Science Foundation of China/ ; 32372528//National Natural Science Foundation of China/ ; TJ-2023-038//Young Elite Scientists Sponsorship Program by Jiangsu Association for Science and Technology/ ; BK20211213//Natural Science Foundation of Jiangsu Province/ ; }, abstract = {The two-spotted spider mite (Tetranychus urticae) is one of the most well-known pesticide-resistant agricultural pests, with resistance often attributed to changes such as target-site mutations and detoxification activation. Recent studies show that pesticide resistance can also be influenced by symbionts, but their involvement in this process in spider mites remains uncertain. Here, we found that infection with Wolbachia, a well-known bacterial reproductive manipulator, significantly increased mite survival after exposure to the insecticides abamectin, cyflumetofen, and pyridaben. Wolbachia-infected (WI) mites showed higher expression of detoxification genes such as P450, glutathione-S-transferase (GST), ABC transporters, and carboxyl/cholinesterases. RNA interference experiments confirmed the role of the two above-mentioned detoxification genes, TuCYP392D2 and TuGSTd05, in pesticide resistance. Increased GST activities were also observed in abamectin-treated WI mites. In addition, when wild populations were treated with abamectin, WI mites generally showed better survival than uninfected mites. However, genetically homogeneous mites with different Wolbachia strains showed similar survival. Finally, abamectin treatment increased Wolbachia abundance without altering the mite's bacterial community. This finding highlights the role of Wolbachia in orchestrating pesticide resistance by modulating host detoxification. By unraveling the intricate interplay between symbionts and pesticide resistance, our study lays the groundwork for pioneering strategies to combat agricultural pests.}, } @article {pmid38381797, year = {2024}, author = {Mendoza-Roldan, JA and Perles, L and Filippi, E and Szafranski, N and Montinaro, G and Carbonara, M and Scalera, R and de Abreu Teles, PP and Walochnik, J and Otranto, D}, title = {Parasites and microorganisms associated with the snakes collected for the "festa Dei serpari" in Cocullo, Italy.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {2}, pages = {e0011973}, doi = {10.1371/journal.pntd.0011973}, pmid = {38381797}, issn = {1935-2735}, abstract = {While in much of the Western world snakes are feared, in the small, rural, mountainous town of Cocullo, in the middle of central Italy, snakes are annually collected and celebrated in a sacro-profane ritual. Every 1st of May, Serpari (snake catchers) capture and showcase dozens of non-venomous snakes to celebrate the ritual of San Domenico. In order to detect potential zoonotic pathogens within this unique epidemiological context, parasites and microorganisms of snakes harvested for the "festa dei serpari" ritual was investigated. Snakes (n = 112) were examined and ectoparasites collected, as well as blood and feces sampled. Ectoparasites were identified morpho-molecularly, and coprological examination conducted through direct smear and flotation. Molecular screenings were performed to identify parasites and microorganisms in collected samples (i.e., Mesostigmata mites, Anaplasma/Ehrlichia spp., Rickettsia spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Babesia/Theileria spp., Cryptosporidium spp., Giardia spp., Leishmania spp. and helminths). Overall, 28.5% (32/112) of snakes were molecularly positive for at least one parasite and/or microorganism. Endosymbiont Wolbachia bacteria were identified from Macronyssidae mites and zoonotic vector-borne bacteria (e.g., Rickettsia, Leishmania), as well as orally transmitted pathogens (i.e., Cryptosporidium, Giardia, Proteus vulgaris, Pseudomonas), were detected from blood and feces. Thus, given the central role of the snakes in the tradition of Cocullo, surveys of their parasitic fauna and associated zoonotic pathogens may aid to generate conservation policies to benefit the human-snake interactions, whilst preserving the cultural patrimony of this event.}, } @article {pmid38380092, year = {2024}, author = {Pramono, AK and Hidayanti, AK and Tagami, Y and Ando, H}, title = {Bacterial community and genome analysis of cytoplasmic incompatibility-inducing Wolbachia in American serpentine leafminer, Liriomyza trifolii.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1304401}, pmid = {38380092}, issn = {1664-302X}, abstract = {Liriomyza trifolii, an agricultural pest, is occasionally infected by Wolbachia. A Wolbachia strain present in Liriomyza trifolii is associated with cytoplasmic incompatibility (CI) effects, leading to the death of embryos resulting from incompatible crosses between antibiotic-treated or naturally Wolbachia-free strain females and Wolbachia-infected males. In this study, high-throughput sequencing of hypervariable rRNA genes was employed to characterize the bacterial community in Wolbachia-infected L. trifolii without antibiotic treatment. The analysis revealed that Wolbachia dominates the bacterial community in L. trifolii, with minor presence of Acinetobacter, Pseudomonas, and Limnobacter. To elucidate the genetic basis of the CI phenotype, metagenomic sequencing was also conducted to assemble the genome of the Wolbachia strain. The draft-genome of the Wolbachia strain wLtri was 1.35 Mbp with 34% GC content and contained 1,487 predicted genes. Notably, within the wLtri genome, there are three distinct types of cytoplasmic incompatibility factor (cif) genes: Type I, Type III, and Type V cifA;B. These genes are likely responsible for inducing the strong cytoplasmic incompatibility observed in L. trifolii.}, } @article {pmid38374896, year = {2024}, author = {Mazel, F and Pitteloud, C and Guisan, A and Pellissier, L}, title = {Contrasted host specificity of gut and endosymbiont bacterial communities in alpine grasshoppers and crickets.}, journal = {ISME communications}, volume = {4}, number = {1}, pages = {ycad013}, pmid = {38374896}, issn = {2730-6151}, abstract = {Bacteria colonize the body of macroorganisms to form associations ranging from parasitic to mutualistic. Endosymbiont and gut symbiont communities are distinct microbiomes whose compositions are influenced by host ecology and evolution. Although the composition of horizontally acquired symbiont communities can correlate to host species identity (i.e. harbor host specificity) and host phylogeny (i.e. harbor phylosymbiosis), we hypothesize that the microbiota structure of vertically inherited symbionts (e.g. endosymbionts like Wolbachia) is more strongly associated with the host species identity and phylogeny than horizontally acquired symbionts (e.g. most gut symbionts). Here, using 16S metabarcoding on 336 guts from 24 orthopteran species (grasshoppers and crickets) in the Alps, we observed that microbiota correlated to host species identity, i.e. hosts from the same species had more similar microbiota than hosts from different species. This effect was ~5 times stronger for endosymbionts than for putative gut symbionts. Although elevation correlated with microbiome composition, we did not detect phylosymbiosis for endosymbionts and putative gut symbionts: closely related host species did not harbor more similar microbiota than distantly related species. Our findings indicate that gut microbiota of studied orthopteran species is more correlated to host identity and habitat than to the host phylogeny. The higher host specificity in endosymbionts corroborates the idea that-everything else being equal-vertically transmitted microbes harbor stronger host specificity signal, but the absence of phylosymbiosis suggests that host specificity changes quickly on evolutionary time scales.}, } @article {pmid38362501, year = {2024}, author = {Hegde, S and Marriott, AE and Pionnier, N and Steven, A and Bulman, C and Gunderson, E and Vogel, I and Koschel, M and Ehrens, A and Lustigman, S and Voronin, D and Tricoche, N and Hoerauf, A and Hübner, MP and Sakanari, J and Aljayyoussi, G and Gusovsky, F and Dagley, J and Hong, DW and O'Neill, P and Ward, SA and Taylor, MJ and Turner, JD}, title = {Combinations of the azaquinazoline anti-Wolbachia agent, AWZ1066S, with benzimidazole anthelmintics synergise to mediate sub-seven-day sterilising and curative efficacies in experimental models of filariasis.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1346068}, doi = {10.3389/fmicb.2024.1346068}, pmid = {38362501}, issn = {1664-302X}, abstract = {Lymphatic filariasis and onchocerciasis are two major neglected tropical diseases that are responsible for causing severe disability in 50 million people worldwide, whilst veterinary filariasis (heartworm) is a potentially lethal parasitic infection of companion animals. There is an urgent need for safe, short-course curative (macrofilaricidal) drugs to eliminate these debilitating parasite infections. We investigated combination treatments of the novel anti-Wolbachia azaquinazoline small molecule, AWZ1066S, with benzimidazole drugs (albendazole or oxfendazole) in up to four different rodent filariasis infection models: Brugia malayi-CB.17 SCID mice, B. malayi-Mongolian gerbils, B. pahangi-Mongolian gerbils, and Litomosoides sigmodontis-Mongolian gerbils. Combination treatments synergised to elicit threshold (>90%) Wolbachia depletion from female worms in 5 days of treatment, using 2-fold lower dose-exposures of AWZ1066S than monotherapy. Short-course lowered dose AWZ1066S-albendazole combination treatments also delivered partial adulticidal activities and/or long-lasting inhibition of embryogenesis, resulting in complete transmission blockade in B. pahangi and L. sigmodontis gerbil models. We determined that short-course AWZ1066S-albendazole co-treatment significantly augmented the depletion of Wolbachia populations within both germline and hypodermal tissues of B. malayi female worms and in hypodermal tissues in male worms, indicating that anti-Wolbachia synergy is not limited to targeting female embryonic tissues. Our data provides pre-clinical proof-of-concept that sub-seven-day combinations of rapid-acting novel anti-Wolbachia agents with benzimidazole anthelmintics are a promising curative and transmission-blocking drug treatment strategy for filarial diseases of medical and veterinary importance.}, } @article {pmid38348307, year = {2023}, author = {Yang, K and Zhang, HY and Wang, P and Jin, GX and Chu, D}, title = {Both symbionts and environmental factors contribute to shape the microbiota in a pest insect, Sogatella furcifera.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1336345}, pmid = {38348307}, issn = {1664-302X}, abstract = {INTRODUCTION: Bacterial symbionts are prevalent in arthropods globally and play a vital role in the fitness and resistance of hosts. While several symbiont infections have been identified in the white-backed planthopper Sogatella furcifera, the impact of environmental factors on the microbiota within S. furcifera remains elusive.

METHODS: In this study, a total of 142 S. furcifera individuals from 18 populations were collected from 14 locations across six countries (China, Thailand, Myanmar, Cambodia, Vietnam, and Laos) analyzed with 2bRAD-M sequencing, to examine the effects of symbionts on the microbiota in the S. furcifera population, as well as the vital effects of environmental factors on the bacterial communities.

RESULTS AND DISCUSSION: Based on the results, in S. furcifera, the presence of symbionts Wolbachia and Cardinium negatively influenced the abundance of other bacteria, including Enterobacter, Acinetobacter, and Lysinibacillus, while Wolbachia infection significantly decreased the diversity of the microbial community. Moreover, several environmental factors, including longitude, latitude, temperature, and precipitation, affected the abundance of symbionts and microbiota diversity in S. furcifera. These results collectively highlight the vital role of Wolbachia in S. furcifera microbiota, as well as the intricate effects of environmental factors on the bacterial communities of S. furcifera.}, } @article {pmid38342109, year = {2024}, author = {Lim, JT and Bansal, S and Chong, CS and Dickens, B and Ng, Y and Deng, L and Lee, C and Tan, LY and Chain, G and Ma, P and Sim, S and Tan, CH and Cook, AR and Ng, LC}, title = {Efficacy of Wolbachia-mediated sterility to reduce the incidence of dengue: a synthetic control study in Singapore.}, journal = {The Lancet. Microbe}, volume = {}, number = {}, pages = {}, doi = {10.1016/S2666-5247(23)00397-X}, pmid = {38342109}, issn = {2666-5247}, abstract = {BACKGROUND: Due to the absence of available therapeutics and good vaccines, vector control solutions are needed to mitigate the spread of dengue. Matings between male Aedes aegypti mosquitoes infected with the wAlbB strain of Wolbachia and wildtype females yield non-viable eggs. We evaluated the efficacy of releasing wAlbB-infected A aegypti male mosquitoes to suppress dengue incidence.

METHODS: In this synthetic control study, we conducted large-scale field trials in Singapore involving release of wAlbB-infected A aegypti male mosquitoes for dengue control via vector population suppression, from epidemiological week (EW) 27, 2018, to EW 26, 2022. We selected two large towns (Yishun and Tampines) to adopt an expanding release strategy and two smaller towns (Bukit Batok and Choa Chu Kang) to adopt a targeted-release approach. Releases were conducted two times a week in high-rise public housing estates. All intervention and control locations practised the same baseline dengue control protocol. The main outcome was weekly dengue incidence rate caused by any dengue virus serotype. We used incidence data collected by the Singapore Ministry of Health to assess the efficacy of the interventions. To compare interventions, we used the synthetic control method to generate appropriate counterfactuals for the intervention towns using a weighted combination of 30 control towns between EW 1, 2014 and EW 26, 2022.

FINDINGS: Our study comprised an at-risk population of 607 872 individuals living in intervention sites and 3 894 544 individuals living in control sites. Interventions demonstrated up to 77·28% (121/156, 95% CI 75·81-78·58) intervention efficacy despite incomplete coverage across all towns until EW 26, 2022. Intervention efficacies increased as release coverage increased across all intervention sites. Releases led to 2242 (95% CI 2092-2391) fewer cases per 100 000 people in intervention sites during the study period. Secondary analysis showed that these intervention effects were replicated across all age groups and both sexes for intervention sites.

INTERPRETATION: Our results demonstrated the potential of Wolbachia-mediated incompatible insect technique for strengthening dengue control in tropical cities, where dengue burden is the greatest.

FUNDING: Singapore Ministry of Finance, Ministry of Sustainability, and the National Environment Agency, and the Singapore National Robotics Program.}, } @article {pmid38327789, year = {2024}, author = {Hoffmann, AA and Ahmad, NW and Keong, WM and Ling, CY and Ahmad, NA and Golding, N and Tierney, N and Jelip, J and Putit, PW and Mokhtar, N and Sandhu, SS and Ming, LS and Khairuddin, K and Denim, K and Rosli, NM and Shahar, H and Omar, T and Ridhuan Ghazali, MK and Aqmar Mohd Zabari, NZ and Abdul Karim, MA and Saidin, MI and Mohd Nasir, MN and Aris, T and Sinkins, SP}, title = {Introduction of Aedes aegypti mosquitoes carrying wAlbB Wolbachia sharply decreases dengue incidence in disease hotspots.}, journal = {iScience}, volume = {27}, number = {2}, pages = {108942}, doi = {10.1016/j.isci.2024.108942}, pmid = {38327789}, issn = {2589-0042}, abstract = {Partial replacement of resident Aedes aegypti mosquitoes with introduced mosquitoes carrying certain strains of inherited Wolbachia symbionts can result in transmission blocking of dengue and other viruses of public health importance. Wolbachia strain wAlbB is an effective transmission blocker and stable at high temperatures, making it particularly suitable for hot tropical climates. Following trial field releases in Malaysia, releases using wAlbB Ae. aegypti have become operationalized by the Malaysian health authorities. We report here on an average reduction in dengue fever of 62.4% (confidence intervals 50-71%) in 20 releases sites when compared to 76 control sites in high-rise residential areas. Importantly the level of dengue reduction increased with Wolbachia frequency, with 75.8% reduction (61-87%) estimated at 100% Wolbachia frequency. These findings indicate large impacts of wAlbB Wolbachia invasions on dengue fever incidence in an operational setting, with incidence expected to further decrease as wider areas are invaded.}, } @article {pmid38327358, year = {2023}, author = {Liu, Y and Zhao, L and Qiu, Z and Yuan, H}, title = {The gut microbiota diversity of five Orthoptera (Insecta, Polyneoptera) insects determined by DNA metabarcoding.}, journal = {Biodiversity data journal}, volume = {11}, number = {}, pages = {e98162}, doi = {10.3897/BDJ.11.e98162}, pmid = {38327358}, issn = {1314-2828}, abstract = {Most orthopteran insects are phytophagous and some are important pests in agriculture and forests. Many intestinal microflora of Orthoptera insects have been reported, primarily from Acridoidea and there have been few reports of other taxa. In this study, we collected 15 individuals representing five species (Ruspolialineosa, Tetrixjaponica, Erianthusversicolor, Gryllotalpaorientalis and Teleogryllusemma) belonging to five orthopteran superfamilies (Tettigonioidea, Tetrigoidea, Eumastacoidea, Gryllotalpoidea and Grylloidea) to characterise and compare the gut microbiota with greater taxonomic width by performing sequencing analysis of the 16S rRNA V4 region in gut material. A total of 606,053 high-quality sequences and 3,105 OTUs were acquired from 15 gut samples representing 24 phyla, 48 classes, 69 orders, 133 families and 219 genera. Firmicutes and bacteria were the most abundant phyla, followed by Bacteroidetes, Cyanobacteria, Actinobacteria and Acidobacteria. At the genus level, Serratia, Citrobacter, Wolbachia, Lactobacillus and Parabacteroides were the most predominant genera in R.lineosa, T.japonica, E.versicolor, G.orientalis and T.emma, respectively. Both Principal Coordinates Analysis (PCoA) and heatmap results revealed significant differences in bacterial community composition across species. Additionally, alpha diversity analysis indicated the bacterial richness was significantly different amongst the five species.}, } @article {pmid38315724, year = {2024}, author = {Namias, A and Ngaku, A and Makoundou, P and Unal, S and Sicard, M and Weill, M}, title = {Intra-lineage microevolution of Wolbachia leads to the emergence of new cytoplasmic incompatibility patterns.}, journal = {PLoS biology}, volume = {22}, number = {2}, pages = {e3002493}, doi = {10.1371/journal.pbio.3002493}, pmid = {38315724}, issn = {1545-7885}, abstract = {Mosquitoes of the Culex pipiens complex are worldwide vectors of arbovirus, filarial nematodes, and avian malaria agents. In these hosts, the endosymbiotic bacteria Wolbachia induce cytoplasmic incompatibility (CI), i.e., reduced embryo viability in so-called incompatible crosses. Wolbachia infecting Culex pipiens (wPip) cause CI patterns of unparalleled complexity, associated with the amplification and diversification of cidA and cidB genes, with up to 6 different gene copies described in a single wPip genome. In wPip, CI is thought to function as a toxin-antidote (TA) system where compatibility relies on having the right antidotes (CidA) in the female to bind and neutralize the male's toxins (CidB). By repeating crosses between Culex isofemale lines over a 17 years period, we documented the emergence of a new compatibility type in real time and linked it to a change in cid genes genotype. We showed that loss of specific cidA gene copies in some wPip genomes results in a loss of compatibility. More precisely, we found that this lost antidote had an original sequence at its binding interface, corresponding to the original sequence at the toxin's binding interface. We showed that these original cid variants are recombinant, supporting a role for recombination rather than point mutations in rapid CI evolution. These results strongly support the TA model in natura, adding to all previous data acquired with transgenes expression.}, } @article {pmid38296097, year = {2024}, author = {Li, JH and Liu, XH and Liang, GR and Gao, HT and Guo, SH and Zhou, XY and Xing, D and Zhao, T and Li, CX}, title = {Microplastics affect mosquito from aquatic to terrestrial lifestyles and are transferred to mammals through mosquito bites.}, journal = {The Science of the total environment}, volume = {}, number = {}, pages = {170547}, doi = {10.1016/j.scitotenv.2024.170547}, pmid = {38296097}, issn = {1879-1026}, abstract = {Microplastics (MPs) transfer from the environment to living organisms is a nonignorable global problem. As a complete metamorphosis insect, the larvae and adult Culex quinquefasciatus mosquito live in aquatic and terrestrial environments, respectively, where they easily access MPs. However, little is known about mosquitoes' potential role in MPs accumulation throughout ecosystems. Therefore, we conducted a study with different MPs particle sizes (0.1/1/10 μm) and concentrations (0.5/5/50 μg/mL) on Cx. quinquefasciatus to address this issue. Once exposed at the young larval stage, MPs could accompany the mosquitoes their entire life. The fluorescence signals of MPs in the larvae were mainly located in the intestines. Its intensity increased (from 3.72 × 10[6] AU to 5.45 × 10[7] AU) as the concentrations of MPs increases. The fluorescence signals of MPs were also detected in the blood and skin tissues of mice bitten by adult mosquitoes with MPs containing in their bodies. Mosquitos exposed to MPs showed longer larval pupation and eclosion time as well as lower adult body weight. In addition, MPs significantly reduced the lethal effect of pyrethroid insecticides (97.77 % vs. 48.88 %, p < 0.05) with 15.1 % removal of the deltamethrin concentration. After MPs exposure, the relative abundance of the Cx. quinquefasciatus gut microbiome, such as Wolbachia spp., Elizabethkingia spp., and Asaia spp., changed as the MPs size and concentration changes. Mosquitoes provide a new pathway for MPs accumulation and transfer to higher-level living organisms. Moreover, MPs significantly reduce the control effect of deltamethrin, providing new guidelines for mosquito insecticide application in MPs contamination circumstances.}, } @article {pmid38287029, year = {2024}, author = {McNamara, CJ and Ant, TH and Harvey-Samuel, T and White-Cooper, H and Martinez, J and Alphey, L and Sinkins, SP}, title = {Transgenic expression of cif genes from Wolbachia strain wAlbB recapitulates cytoplasmic incompatibility in Aedes aegypti.}, journal = {Nature communications}, volume = {15}, number = {1}, pages = {869}, pmid = {38287029}, issn = {2041-1723}, support = {202888/Z/16/Z//Wellcome Trust (Wellcome)/ ; MC_ST_CVR_2019//RCUK | Medical Research Council (MRC)/ ; }, abstract = {The endosymbiotic bacteria Wolbachia can invade insect populations by modifying host reproduction through cytoplasmic incompatibility (CI), an effect that results in embryonic lethality when Wolbachia-carrying males mate with Wolbachia-free females. Here we describe a transgenic system for recreating CI in the major arbovirus vector Aedes aegypti using CI factor (cif) genes from wAlbB, a Wolbachia strain currently being deployed to reduce dengue transmission. CI-like sterility is induced when cifA and cifB are co-expressed in testes; this sterility is rescued by maternal cifA expression, thereby reproducing the pattern of Wolbachia-induced CI. Expression of cifB alone is associated with extensive DNA damage and disrupted spermatogenesis. The strength of rescue by maternal cifA expression is dependent on the comparative levels of cifA/cifB expression in males. These findings are consistent with CifB acting as a toxin and CifA as an antitoxin, with CifA attenuating CifB toxicity in both the male germline and in developing embryos. These findings provide important insights into the interactions between cif genes and their mechanism of activity and provide a foundation for the building of a cif gene-based drive system in Ae. aegypti.}, } @article {pmid38278818, year = {2024}, author = {Piccinno, R and Tatti, A and Avosani, S and Galla, G and Lazazzara, V and Pedrazzoli, F and Zadra, N and Rodeghiero, M and Seljak, G and Özgen, İ and Hauffe, HC and Verrastro, V and Stacconi, MVR and Mazzoni, V and Rota-Stabelli, O}, title = {A multidisciplinary approach to tackling invasive species: barcoding, morphology, and metataxonomy of the leafhopper Arboridia adanae.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {2229}, pmid = {38278818}, issn = {2045-2322}, abstract = {The leafhopper genus Arboridia includes several species that feed on Vitis vinifera and cause leaf chlorosis. We report the first alien Arboridia infestation in Italy in 2021 in an Apulian vineyard. To confirm the taxonomic status of the species responsible for crop damage, and reconstruct its demographic history, we barcoded individuals from Apulia together with Arboridia spp. from Crete (Greece), A. adanae from Central Turkey and other specimens of the presumed sister species, A. dalmatina from Dalmatia (Croatia). Molecular phylogenies and barcoding gap analysis identified clades not associated with sampling locations. This result is incongruent with classical specimen assignment and is further supported by morphological analyses, which did not reveal significant differences among the populations. Therefore, we propose A. dalmatina as a junior synonym of A. adanae, which would become the only grapevine-related Arboridia species in the eastern Mediterranean. To further characterise A. adanae evolution, we performed a molecular clock analysis that suggested a radiation during the Pleistocene glaciations. Finally, to assess whether the Apulian individuals carried microorganisms of agricultural relevance, we sequenced their bacterial microbiota using 16S rRNA amplicon sequencing identifying three phytopathogens not generally associated with Arboridia activities as well as Wolbachia in one Apulian haplogroup. We discuss the agricultural implications of this infestation.}, } @article {pmid38257815, year = {2024}, author = {Rainey, SM and Lefteri, DA and Darby, C and Kohl, A and Merits, A and Sinkins, SP}, title = {Evidence of Differences in Cellular Regulation of Wolbachia-Mediated Viral Inhibition between Alphaviruses and Flaviviruses.}, journal = {Viruses}, volume = {16}, number = {1}, pages = {}, doi = {10.3390/v16010115}, pmid = {38257815}, issn = {1999-4915}, support = {202888/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; WT094664MA/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, abstract = {The intracellular bacterium Wolbachia is increasingly being utilised in control programs to limit the spread of arboviruses by Aedes mosquitoes. Achieving a better understanding of how Wolbachia strains can reduce viral replication/spread could be important for the long-term success of such programs. Previous studies have indicated that for some strains of Wolbachia, perturbations in lipid metabolism and cholesterol storage are vital in Wolbachia-mediated antiviral activity against the flaviviruses dengue and Zika; however, it has not yet been examined whether arboviruses in the alphavirus group are affected in the same way. Here, using the reporters for the alphavirus Semliki Forest virus (SFV) in Aedes albopictus cells, we found that Wolbachia strains wMel, wAu and wAlbB blocked viral replication/translation early in infection and that storage of cholesterol in lipid droplets is not key to this inhibition. Another alphavirus, o'nyong nyong virus (ONNV), was tested in both Aedes albopictus cells and in vivo in stable, transinfected Aedes aegypti mosquito lines. The strains wMel, wAu and wAlbB show strong antiviral activity against ONNV both in vitro and in vivo. Again, 2-hydroxypropyl-β-cyclodextrin (2HPCD) was not able to rescue ONNV replication in cell lines, suggesting that the release of stored cholesterol caused by wMel is not able to rescue blockage of ONNV. Taken together, this study shows that alphaviruses appear to be inhibited early in replication/translation and that there may be differences in how alphaviruses are inhibited by Wolbachia in comparison to flaviviruses.}, } @article {pmid38254452, year = {2024}, author = {Cheng, Z and Liu, Q and Huang, X}, title = {Partial Correspondence between Host Plant-Related Differentiation and Symbiotic Bacterial Community in a Polyphagous Insect.}, journal = {Animals : an open access journal from MDPI}, volume = {14}, number = {2}, pages = {}, doi = {10.3390/ani14020283}, pmid = {38254452}, issn = {2076-2615}, support = {31970446//National Natural Science Foundation of China/ ; }, abstract = {Host plants play a vital role in insect population differentiation, while symbiotic associations between bacteria and insects are ubiquitous in nature. However, existing studies have given limited attention to the connection between host-related differentiation and symbiotic bacterial communities in phytophagous insects. In this study, we collected 58 samples of Aphis odinae from different host plants in southern China and constructed phylogenetic trees to investigate their differentiation in relation to host plants. We also selected aphid samples from the five most preferred host plants and analyzed their symbiotic bacterial composition using Illumina sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. The phylogeny and symbiotic bacterial community structure of A. odinae populations on different host plants showed that samples from Triadica sebifera (Euphorbiaceae) had a consistent presence of Wolbachia as the predominant secondary symbiont and suggested the possibility of undergoing differentiation. Conversely, although differentiation was observed in samples from Rhus chinensis (Anacardiaceae), no consistent presence of predominant secondary symbionts was found. Additionally, the samples from Heptapleurum heptaphyllum (Araliaceae) consistently carried Serratia, but no host differentiation was evident. In summary, this study reveals a partial correspondence between symbiotic bacterial communities and host-related differentiation in A. odinae. The findings contribute to our understanding of the microevolutionary influencing the macroevolutionary relationships between bacterial symbionts and phytophagous insects. The identification of specific symbionts associated with host-related differentiation provides valuable insights into the intricate dynamics of insect-bacteria interactions.}, } @article {pmid38243558, year = {2024}, author = {Sohail, A and Anders, KL and McGuinness, SL and Leder, K}, title = {The epidemiology of imported and locally-acquired dengue in Australia, 2012-2022.}, journal = {Journal of travel medicine}, volume = {}, number = {}, pages = {}, doi = {10.1093/jtm/taae014}, pmid = {38243558}, issn = {1708-8305}, support = {2002792//National Health and Medical Research/ ; 2017229//NHMRC Investigator Grants/ ; 1155005//NHMRC Senior Research Fellowship/ ; 224459/Z/21/Z//Welcome Trust/ ; }, abstract = {BACKGROUND: Dengue is the most important arboviral disease globally, and poses ongoing challenges for control including in non-endemic countries with competent mosquito vectors at risk of local transmission through imported cases. We examined recent epidemiological trends in imported and locally-acquired dengue in Australia, where the Wolbachia mosquito population replacement method was implemented throughout dengue-prone areas of northern Queensland between 2011-2019.

METHODS: We analysed dengue cases reported to the Australian National Notifiable Disease Surveillance System (NNDSS) between January 2012 and December 2022, and Australian traveller movement data.

RESULTS: Between 2012 and 2022, 13 343 dengue cases were reported in Australia (median 1466 annual cases); 12 568 cases (94.2%) were imported, 584 (4.4%) were locally acquired and 191 (1.4%) had no origin recorded. Locally-acquired cases decreased from a peak in 2013 (n = 236) to zero in 2021-2022. Annual incidence of imported dengue ranged from 8·29/100000 (n = 917 cases) to 22·10/100000 (n = 2203) annual traveller movements between 2012 and 2019, decreased in 2020 (6·74/100000 traveller movements; n = 191) and 2021 (3·32/100000 traveller movements; n = 10) during COVID-19-related border closures, then rose to 34·79/100000 traveller movements (n = 504) in 2022. Imported cases were primarily acquired in Southeast Asia (n = 9323; 74%), Southern and Central Asia (n = 1555; 12%) and Oceania (n = 1341; 11%). Indonesia (n = 5778; 46%) and Thailand (n = 1483; 12%) were top acquisition countries. DENV-2 (n = 2147; 42%) and DENV-1 (n = 1526; 30%) were predominant serotypes.

CONCLUSION: Our analysis highlights Australia's successful control of locally-acquired dengue with Wolbachia. Imported dengue trends reflect both Australian travel destinations and patterns and local epidemiology in endemic countries.

FUNDING: AS is supported by a National Health and Medical Research (NHMRC) Postgraduate Scholarship (grant number 2002792), SLM by NHMRC Investigator Grants (grant number 2017229), KL by a NHMRC Senior Research Fellowship (grant number 1155005), and KLA by the Welcome Trust (grant number 224459/Z/21/Z).}, } @article {pmid38235434, year = {2023}, author = {Mantilla-Granados, JS and Castellanos, JE and Velandia-Romero, ML}, title = {A tangled threesome: understanding arbovirus infection in Aedes spp. and the effect of the mosquito microbiota.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1287519}, pmid = {38235434}, issn = {1664-302X}, abstract = {Arboviral infections transmitted by Aedes spp. mosquitoes are a major threat to human health, particularly in tropical regions but are expanding to temperate regions. The ability of Aedes aegypti and Aedes albopictus to transmit multiple arboviruses involves a complex relationship between mosquitoes and the virus, with recent discoveries shedding light on it. Furthermore, this relationship is not solely between mosquitoes and arboviruses, but also involves the mosquito microbiome. Here, we aimed to construct a comprehensive review of the latest information about the arbovirus infection process in A. aegypti and A. albopictus, the source of mosquito microbiota, and its interaction with the arbovirus infection process, in terms of its implications for vectorial competence. First, we summarized studies showing a new mechanism for arbovirus infection at the cellular level, recently described innate immunological pathways, and the mechanism of adaptive response in mosquitoes. Second, we addressed the general sources of the Aedes mosquito microbiota (bacteria, fungi, and viruses) during their life cycle, and the geographical reports of the most common microbiota in adults mosquitoes. How the microbiota interacts directly or indirectly with arbovirus transmission, thereby modifying vectorial competence. We highlight the complexity of this tripartite relationship, influenced by intrinsic and extrinsic conditions at different geographical scales, with many gaps to fill and promising directions for developing strategies to control arbovirus transmission and to gain a better understanding of vectorial competence. The interactions between mosquitoes, arboviruses and their associated microbiota are yet to be investigated in depth.}, } @article {pmid38230418, year = {2024}, author = {Nindrea, RD}, title = {Wolbachia: New Hopes for the Prevention of Dengue Hemorrhagic Fever in Indonesia.}, journal = {Asia-Pacific journal of public health}, volume = {}, number = {}, pages = {10105395231226296}, doi = {10.1177/10105395231226296}, pmid = {38230418}, issn = {1941-2479}, } @article {pmid38204789, year = {2024}, author = {Arai, H and Watada, M and Kageyama, D}, title = {Two male-killing Wolbachia from Drosophila birauraia that are closely related but distinct in genome structure.}, journal = {Royal Society open science}, volume = {11}, number = {1}, pages = {231502}, pmid = {38204789}, issn = {2054-5703}, abstract = {Insects harbour diverse maternally inherited bacteria and viruses, some of which have evolved to kill the male progeny of their hosts (male killing: MK). The fly species Drosophila biauraria carries a maternally transmitted MK-inducing partiti-like virus, but it was unknown if it carries other MK-inducing endosymbionts. Here, we identified two male-killing Wolbachia strains (wBiau1 and wBiau2) from D. biauraria and compared their genomes to elucidate their evolutionary processes. The two strains were genetically closely related but had exceptionally different genome structures with considerable rearrangements compared with combinations of other Wolbachia strains. Despite substantial changes in the genome structure, the two Wolbachia strains did not experience gene losses that would disrupt the male-killing expression or persistence in the host population. The two Wolbachia-infected matrilines carried distinct mitochondrial haplotypes, suggesting that wBiau1 and wBiau2 have invaded D. biauraria independently and undergone considerable genome changes owing to unknown selective pressures in evolutionary history. This study demonstrated the presence of three male-killers from two distinct origins in one fly species and highlighted the diverse and rapid genome evolution of MK Wolbachia in the host.}, } @article {pmid38196945, year = {2023}, author = {Côrtes, N and Lira, A and Prates-Syed, W and Dinis Silva, J and Vuitika, L and Cabral-Miranda, W and Durães-Carvalho, R and Balan, A and Cabral-Marques, O and Cabral-Miranda, G}, title = {Integrated control strategies for dengue, Zika, and Chikungunya virus infections.}, journal = {Frontiers in immunology}, volume = {14}, number = {}, pages = {1281667}, pmid = {38196945}, issn = {1664-3224}, mesh = {Animals ; Humans ; *Zika Virus ; *Chikungunya Fever/epidemiology/prevention & control ; Mosquito Vectors ; *Zika Virus Infection/epidemiology/prevention & control ; Antiviral Agents ; *Dengue/epidemiology/prevention & control ; }, abstract = {Arboviruses are a major threat to public health in tropical regions, encompassing over 534 distinct species, with 134 capable of causing diseases in humans. These viruses are transmitted through arthropod vectors that cause symptoms such as fever, headache, joint pains, and rash, in addition to more serious cases that can lead to death. Among the arboviruses, dengue virus stands out as the most prevalent, annually affecting approximately 16.2 million individuals solely in the Americas. Furthermore, the re-emergence of the Zika virus and the recurrent outbreaks of chikungunya in Africa, Asia, Europe, and the Americas, with one million cases reported annually, underscore the urgency of addressing this public health challenge. In this manuscript we discuss the epidemiology, viral structure, pathogenicity and integrated control strategies to combat arboviruses, and the most used tools, such as vaccines, monoclonal antibodies, treatment, etc., in addition to presenting future perspectives for the control of arboviruses. Currently, specific medications for treating arbovirus infections are lacking, and symptom management remains the primary approach. However, promising advancements have been made in certain treatments, such as Chloroquine, Niclosamide, and Isatin derivatives, which have demonstrated notable antiviral properties against these arboviruses in vitro and in vivo experiments. Additionally, various strategies within vector control approaches have shown significant promise in reducing arbovirus transmission rates. These encompass public education initiatives, targeted insecticide applications, and innovative approaches like manipulating mosquito bacterial symbionts, such as Wolbachia. In conclusion, combatting the global threat of arbovirus diseases needs a comprehensive approach integrating antiviral research, vaccination, and vector control. The continued efforts of research communities, alongside collaborative partnerships with public health authorities, are imperative to effectively address and mitigate the impact of these arboviral infections on public health worldwide.}, } @article {pmid38193019, year = {2024}, author = {Mat Udin, AS and Uni, S and Rodrigues, J and Martin, C and Junker, K and Agatsuma, T and Low, VL and Saijuntha, W and Omar, H and Zainuri, NA and Fukuda, M and Matsubayashi, M and Kimura, D and Takaoka, H and Ramli, R}, title = {Redescription, molecular characterisation and Wolbachia endosymbionts of Mansonella (Tupainema) dunni (Mullin & Orihel, 1972) (Spirurida: Onchocercidae) from the common treeshrew Tupaia glis Diard & Duvaucel (Mammalia: Scandentia) in Peninsular Malaysia.}, journal = {Current research in parasitology & vector-borne diseases}, volume = {5}, number = {}, pages = {100154}, pmid = {38193019}, issn = {2667-114X}, abstract = {The genus Mansonella Faust, 1929 includes 29 species, mainly parasites of platyrrhine monkeys in South America and anthropoid apes in Africa. In Malaysia, Mansonella (Tupainema) dunni (Mullin & Orihel, 1972) was described from the common treeshrew Tupaia glis Diard & Duvaucel (Scandentia). In a recent classification of the genus Mansonella, seven subgenera were proposed, with M. (Tup.) dunni as a monotypic species in the subgenus Tupainema. In this study, we collected new material of M. (Tup.) dunni from common treeshrews in Peninsular Malaysia and redescribed the morphological features of this species. We found that M. (Tup.) dunni differs from M. (Cutifilaria) perforata Uni et al., 2004 from sika deer Cervus nippon (Cetartiodactyla) in Japan, with regards to morphological features and predilection sites in their respective hosts. Based on multi-locus sequence analyses, we examined the molecular phylogeny of M. (Tup.) dunni and its Wolbachia genotype. Species of the genus Mansonella grouped monophyletically in clade ONC5 and M. (Tup.) dunni was placed in the most derived position within this genus. Mansonella (Tup.) dunni was closely related to M. (M.) ozzardi (Manson, 1897) from humans in Central and South America, and most distant from M. (C.) perforata. The calculated p-distances between the cox1 gene sequences for M. (Tup.) dunni and its congeners were 13.09% for M. (M.) ozzardi and 15.6-16.15% for M. (C.) perforata. The molecular phylogeny of Mansonella spp. thus corroborates their morphological differences. We determined that M. (Tup.) dunni harbours Wolbachia endosymbionts of the supergroup F genotype, in keeping with all other Mansonella species screened to date.}, } @article {pmid38192184, year = {2024}, author = {Strunov, A and Schönherr, C and Kapun, M}, title = {Wolbachia effects on thermal preference of natural Drosophila melanogaster are influenced by host genetic background, Wolbachia type, and bacterial titer.}, journal = {Environmental microbiology}, volume = {}, number = {}, pages = {e16579}, doi = {10.1111/1462-2920.16579}, pmid = {38192184}, issn = {1462-2920}, support = {FWF P32275//Austrian Science Fund/ ; }, abstract = {Temperature plays a fundamental role in the fitness of all organisms. In particular, it strongly affects metabolism and reproduction in ectotherms that have limited physiological capabilities to regulate their body temperature. The influence of temperature variation on the physiology and behaviour of ectotherms is well studied but we still know little about the influence of symbiotic interactions on thermal preference (Tp) of the host. A growing number of studies focusing on the Wolbachia-Drosophila host-symbiont system found that Wolbachia can influence Tp in Drosophila laboratory strains. Here, we investigated the effect of Wolbachia on Tp in wild-type D. melanogaster flies recently collected from nature. Consistent with previous data, we found reduced Tp compared to an uninfected control in one of two fly strains infected with the wMelCS Wolbachia type. Additionally, we, for the first time, found that Wolbachia titer variation influences the thermal preference of the host fly. These data indicate that the interaction of Wolbachia and Drosophila resulting in behavioural variation is strongly influenced by the genetic background of the host and symbiont. More studies are needed to better understand the evolutionary significance of Tp variation influenced by Wolbachia in natural Drosophila populations.}, } @article {pmid38172384, year = {2024}, author = {Volpe, HXL and Carmo-Sousa, M and Luvizotto, RAG and de Freitas, R and Esperança, V and Darolt, JC and Pegoraro, AAL and Magalhães, DM and Favaris, AP and Wulff, NA and Miranda, MP and Bento, JMS and Leal, WS}, title = {The greening-causing agent alters the behavioral and electrophysiological responses of the Asian citrus psyllid to a putative sex pheromone.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {455}, pmid = {38172384}, issn = {2045-2322}, support = {201600147//National Institute of Science and Technology of Semiochemicals in Agriculture (INCT)/ ; 2014/50871-0//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 201600147//FUNDECITRUS/ University of California-Davis Research Agreement/ ; }, mesh = {Female ; Male ; Animals ; *Hemiptera/physiology ; *Sex Attractants/pharmacology ; Brazil ; *Citrus/microbiology ; *Citrus sinensis ; *Wolbachia ; *Rhizobiaceae/physiology ; Acetates ; Plant Diseases/microbiology ; }, abstract = {The Asian Citrus Psyllid (ACP), Diaphorina citri, is a vector of the pathological bacterium Candidatus Liberibacter asiaticus (CLas), which causes the most devastating disease to the citrus industry worldwide, known as greening or huanglongbing (HLB). Earlier field tests with an acetic acid-based lure in greening-free, 'Valencia' citrus orange groves in California showed promising results. The same type of lures tested in São Paulo, Brazil, showed unsettling results. During the unsuccessful trials, we noticed a relatively large proportion of females in the field, ultimately leading us to test field-collected males and females for Wolbachia and CLas. The results showed high rates of Wolbachia and CLas infection in field populations. We then compared the olfactory responses of laboratory-raised, CLas-free, and CLas-infected males to acetic acid. As previously reported, CLas-uninfected males responded to acetic acid at 1 µg. Surprisingly, CLas-infected males required 50 × higher doses of the putative sex pheromone, thus explaining the failure to capture CLas-infected males in the field. CLas infection was also manifested in electrophysiological responses. Electroantennogram responses from CLas-infected ACP males were significantly higher than those obtained with uninfected males. To the best of our knowledge, this is the first report of a pathogen infection affecting a vector's response to a sex attractant.}, } @article {pmid38162582, year = {2023}, author = {Muharromah, AF and Reyes, JIL and Kagia, N and Watanabe, K}, title = {Genome-wide detection of Wolbachia in natural Aedes aegypti populations using ddRAD-Seq.}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1252656}, pmid = {38162582}, issn = {2235-2988}, mesh = {Humans ; Animals ; Male ; Female ; *Aedes/microbiology ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Philippines ; Ribosomal Proteins ; }, abstract = {BACKGROUND: Wolbachia, an endosymbiotic bacterium, is globally used to control arboviruses because of its ability to block arboviral replication and manipulate the reproduction of Wolbachia host, Aedes aegypti. Polymerase chain reaction (PCR)-based Wolbachia detection has been recently reported from natural Ae. aegypti populations. However, due to the technical limitations of PCR, such as primer incompatibility, PCR-based assays are not sufficiently reliable or accurate. In this study, we examined double digestion restriction site-associated DNA sequencing (ddRAD-Seq) efficiency and limitations in Wolbachia detection and quantification in field-collected Ae. aegypti natural populations in Metro Manila, the Philippines, compared with PCR-based assays.

METHODS: A total of 217 individuals Ae. aegypti were collected from Metropolitan Manila, Philippines. We separated it into 14 populations consisting of 7 female and male populations. We constructed a library for pool ddRAD-Seq per population and also screened for Wolbachia by PCR assays using wsp and 16S rRNA. Wolbachia density per population were measured using RPS17 as the housekeeping gene.

RESULTS: From 146,239,637 sequence reads obtained, 26,299 and 43,778 reads were mapped across the entire Wolbachia genome (with the wAlbA and wAlbB strains, respectively), suggesting that ddRAD-Seq complements PCR assays and supports more reliable Wolbachia detection from a genome-wide perspective. The number of reads mapped to the Wolbachia genome per population positively correlated with the number of Wolbachia-infected individuals per population based on PCR assays and the relative density of Wolbachia in the Ae. aegypti populations based on qPCR, suggesting ddRAD-Seq-based semi-quantification of Wolbachia by ddRAD-Seq. Male Ae. aegypti exhibited more reads mapped to the Wolbachia genome than females, suggesting higher Wolbachia prevalence rates in their case. We detected 150 single nucleotide polymorphism loci across the Wolbachia genome, allowing for more accurate the detection of four strains: wPip, wRi, TRS of Brugia malayi, and wMel.

CONCLUSIONS: Taken together, our results demonstrate the feasibility of ddRAD-Seq-based Wolbachia detection from field-collected Ae. aegypti mosquitoes.}, } @article {pmid38157329, year = {2023}, author = {, }, title = {Expression of Concern: A Potential Role for the Interaction of Wolbachia Surface Proteins with the Brugia malayi Glycolytic Enzymes and Cytoskeleton in Maintenance of Endosymbiosis.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {12}, pages = {e0011860}, pmid = {38157329}, issn = {1935-2735}, } @article {pmid38143905, year = {2023}, author = {Mouillaud, T and Berger, A and Buysse, M and Rahola, N and Daron, J and Agbor, JP and Sango, SN and Neafsey, DE and Duron, O and Ayala, D}, title = {Limited association between Wolbachia and Plasmodium falciparum infections in natural populations of the major malaria mosquito Anopheles moucheti.}, journal = {Evolutionary applications}, volume = {16}, number = {12}, pages = {1999-2006}, pmid = {38143905}, issn = {1752-4571}, abstract = {Since the discovery of natural malaria vector populations infected by the endosymbiont bacterium Wolbachia, a renewed interest has arisen for using this bacterium as an alternative for malaria control. Among naturally infected mosquitoes, Anopheles moucheti, a major malaria mosquito in Central Africa, exhibits one of the highest prevalences of Wolbachia infection. To better understand whether this maternally inherited bacterium could be used for malaria control, we investigated Wolbachia influence in An. moucheti populations naturally infected by the malaria parasite Plasmodium falciparum. To this end, we collected mosquitoes in a village from Cameroon, Central Africa, where this mosquito is the main malaria vector. We found that the prevalence of Wolbachia bacterium was almost fixed in the studied mosquito population, and was higher than previously recorded. We also quantified Wolbachia in whole mosquitoes and dissected abdomens, confirming that the bacterium is also elsewhere than in the abdomen, but at lower density. Finally, we analyzed the association of Wolbachia presence and density on P. falciparum infection. Wolbachia density was slightly higher in mosquitoes infected with the malaria parasite than in uninfected mosquitoes. However, we observed no correlation between the P. falciparum and Wolbachia densities. In conclusion, our study indicates that naturally occurring Wolbachia infection is not associated to P. falciparum development within An. moucheti mosquitoes.}, } @article {pmid38139239, year = {2023}, author = {Gruntenko, NE and Deryuzhenko, MA and Andreenkova, OV and Shishkina, OD and Bobrovskikh, MA and Shatskaya, NV and Vasiliev, GV}, title = {Drosophila melanogaster Transcriptome Response to Different Wolbachia Strains.}, journal = {International journal of molecular sciences}, volume = {24}, number = {24}, pages = {}, pmid = {38139239}, issn = {1422-0067}, support = {21-14-00090//Russian Science Foundation/ ; }, mesh = {Animals ; Female ; *Drosophila melanogaster/physiology ; *Wolbachia/genetics ; Transcriptome ; Genotype ; }, abstract = {Wolbachia is a maternally inherited, intercellular bacterial symbiont of insects and some other invertebrates. Here, we investigated the effect of two different Wolbachia strains, differing in a large chromosomal inversion, on the differential expression of genes in D. melanogaster females. We revealed significant changes in the transcriptome of the infected flies compared to the uninfected ones, as well as in the transcriptome of flies infected with the Wolbachia strain, wMelPlus, compared to flies infected with the wMelCS[112] strain. We linked differentially expressed genes (DEGs) from two pairwise comparisons, "uninfected-wMelPlus-infected" and "uninfected-wMelCS[112]-infected", into two gene networks, in which the following functional groups were designated: "Proteolysis", "Carbohydrate transport and metabolism", "Oxidation-reduction process", "Embryogenesis", "Transmembrane transport", "Response to stress" and "Alkaline phosphatases". Our data emphasized similarities and differences between infections by different strains under study: a wMelPlus infection results in more than double the number of upregulated DEGs and half the number of downregulated DEGs compared to a wMelCS[112] infection. Thus, we demonstrated that Wolbachia made a significant contribution to differential expression of host genes and that the bacterial genotype plays a vital role in establishing the character of this contribution.}, } @article {pmid38132636, year = {2023}, author = {Loterio, RK and Monson, EA and Templin, R and de Bruyne, JT and Flores, HA and Mackenzie, JM and Ramm, G and Helbig, KJ and Simmons, CP and Fraser, JE}, title = {Antiviral Wolbachia strains associate with Aedes aegypti endoplasmic reticulum membranes and induce lipid droplet formation to restrict dengue virus replication.}, journal = {mBio}, volume = {}, number = {}, pages = {e0249523}, doi = {10.1128/mbio.02495-23}, pmid = {38132636}, issn = {2150-7511}, abstract = {Aedes aegypti transmits a range of important human pathogenic viruses like dengue. However, infection of Ae. aegypti with the insect endosymbiotic bacterium, Wolbachia, reduces the risk of mosquito to human viral transmission. Wolbachia is being utilized at field sites across more than 13 countries to reduce the incidence of viruses like dengue, but it is not well understood how Wolbachia induces its antiviral effects. To examine this at the subcellular level, we compared how different strains of Wolbachia with varying antiviral strengths associate with and modify host cell structures. Strongly antiviral strains were found to specifically associate with the host endoplasmic reticulum and induce striking impacts on host cell lipid droplets. Inhibiting Wolbachia-induced lipid redistribution partially restored dengue virus replication demonstrating this is a contributing role for Wolbachia's antiviral activity. These findings provide new insights into how antiviral Wolbachia strains associate with and modify Ae. aegypti host cells.}, } @article {pmid38132616, year = {2023}, author = {Shapoval, NA and Kir'yanov, AV and Krupitsky, AV and Yakovlev, RV and Romanovich, AE and Zhang, J and Cong, Q and Grishin, NV and Kovalenko, MG and Shapoval, GN}, title = {Phylogeography of Two Enigmatic Sulphur Butterflies, Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897 (Lepidoptera, Pieridae), with Relations to Wolbachia Infection.}, journal = {Insects}, volume = {14}, number = {12}, pages = {}, pmid = {38132616}, issn = {2075-4450}, support = {22-24-01086//Russian Science Foundation/ ; }, abstract = {The genus Colias Fabricius, 1807 includes numerous taxa and forms with uncertain status and taxonomic position. Among such taxa are Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897, interpreted in the literature either as conspecific forms, as subspecies of different but morphologically somewhat similar Colias species or as distinct species-level taxa. Based on mitochondrial and nuclear DNA markers, we reconstructed a phylogeographic pattern of the taxa in question. We recover and include in our analysis DNA barcodes of the century-old type specimens, the lectotype of C. tamerlana deposited in the Natural History Museum (Museum für Naturkunde), Berlin, Germany (ZMHU) and the paralectotype of C. tamerlana and the lectotype of C. mongola deposited in the Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia (ZISP). Our analysis grouped all specimens within four (HP_I-HP_IV) deeply divergent but geographically poorly structured clades which did not support nonconspecifity of C. mongola-C. tamerlana. We also show that all studied females of the widely distributed haplogroup HP_II were infected with a single Wolbachia strain belonging to the supergroup B, while the males of this haplogroup, as well as all other investigated specimens of both sexes, were not infected. Our data highlight the relevance of large-scale sampling dataset analysis and the need for testing for Wolbachia infection to avoid erroneous phylogenetic reconstructions and species misidentification.}, } @article {pmid38132601, year = {2023}, author = {Adonyeva, NV and Efimov, VM and Gruntenko, NE}, title = {The Effect of Genotype Combinations of Wolbachia and Its Drosophila melanogaster Host on Fertility, Developmental Rate and Heat Stress Resistance of Flies.}, journal = {Insects}, volume = {14}, number = {12}, pages = {}, pmid = {38132601}, issn = {2075-4450}, support = {21-14-00090//Russian Science Foundation/ ; }, abstract = {The best-known effect of the intracellular bacterium Wolbachia is its mostly negative influence on the reproduction of the host. However, there is evidence of a positive influence of Wolbachia on the host's resistance to stress, pathogens, and viruses. Here, we analyzed the effects of two Wolbachia strains belonging to wMel and wMelCS genotypes on D. melanogaster traits, such as fertility, survival under acute heat stress, and developmental rate. We found that D. melanogaster lines under study differ significantly in the above-mentioned characteristics, both when the natural infection was preserved, and when it was eliminated. One of Wolbachia strains, wMel, did not affect any of the studied traits. Another strain, wMelPlus, had a significant effect on the development time. Moreover, this effect is observed not only in the line in which it was discovered but also in the one it was transferred to. When transferred to a new line, wMelPlus also caused changes in survival under heat stress. Thus, it could be concluded that Wolbachia-Drosophila interaction depends on the genotypes of both the host and the symbiont, but some Wolbachia effects could depend not on the genotypes, but on the fact of recent transfer of the symbiont.}, } @article {pmid38132578, year = {2023}, author = {Zadra, N and Tatti, A and Silverj, A and Piccinno, R and Devilliers, J and Lewis, C and Arnoldi, D and Montarsi, F and Escuer, P and Fusco, G and De Sanctis, V and Feuda, R and Sánchez-Gracia, A and Rizzoli, A and Rota-Stabelli, O}, title = {Shallow Whole-Genome Sequencing of Aedes japonicus and Aedes koreicus from Italy and an Updated Picture of Their Evolution Based on Mitogenomics and Barcoding.}, journal = {Insects}, volume = {14}, number = {12}, pages = {}, pmid = {38132578}, issn = {2075-4450}, support = {PRIN 2022 Prot. n 2022YNL8ZH//Partially funded by the European Union under NextGenerationEU/ ; }, abstract = {Aedes japonicus and Aedes koreicus are two invasive mosquitoes native to East Asia that are quickly establishing in temperate regions of Europe. Both species are vectors of arboviruses, but we currently lack a clear understanding of their evolution. Here, we present new short-read, shallow genome sequencing of A. japonicus and A. koreicus individuals from northern Italy, which we used for downstream phylogenetic and barcode analyses. We explored associated microbial DNA and found high occurrences of Delftia bacteria in both samples, but neither Asaia nor Wolbachia. We then assembled complete mitogenomes and used these data to infer divergence times estimating the split of A. japonicus from A. koreicus in the Oligocene, which was more recent than that previously reported using mitochondrial markers. We recover a younger age for most other nodes within Aedini and other Culicidae. COI barcoding and phylogenetic analyses indicate that A. japonicus yaeyamensis, A. japonicus amamiensis, and the two A. koreicus sampled from Europe should be considered as separate species within a monophyletic species complex. Our studies further clarify the evolution of A. japonicus and A. koreicus, and indicate the need to obtain whole-genome data from putative species in order to disentangle their complex patterns of evolution.}, } @article {pmid38132308, year = {2023}, author = {Lin, YJ and Yeh, CH and Wu, CZ and Wu, LH}, title = {Climate and Wolbachia Impacts on Anoplolepis gracilipes (Hymenoptera: Formicidae).}, journal = {Biology}, volume = {12}, number = {12}, pages = {}, doi = {10.3390/biology12121482}, pmid = {38132308}, issn = {2079-7737}, support = {111-2313-B-020-003-MY3//National Science and Technology Council, Taiwan/ ; 112AS-5.5.3-BQ-B1//Animal and Plant Health Inspection Agency (APHIA), Ministry of Agriculture (MOA), Taiwan/ ; }, abstract = {The yellow crazy ant (Anoplolepis gracilipes (Smith, 1857)) is a prominent invasive species exhibiting variable population dynamics. Through collecting long-term climate data and validating field surveys with MaxEnt SDM projections, our results indicated that winter temperature and precipitation accumulation strongly influence the population dynamics. An aggression analysis showed that A. gracilipes nests with higher aggression levels (over 2.5 scores) experienced a higher mean maximum temperature (31.84 ± 0.43 °C) and lower prevalence of wAgra (84.8 ± 4.70%) in A. gracilipes from June to October. The nest manipulation and aggression experiments confirmed that temperature increases aggression (1.3 to 2.8 scores) among A. gracilipes workers due to the reduced prevalence of wAgra. To the best of our knowledge, this is the first case of a notable reduction in the prevalence of Wolbachia (100 to 66%) within a colony of A. gracilipes while maintaining stable nests for further experiments.}, } @article {pmid38125201, year = {2024}, author = {Ochieng, FO}, title = {SEIRS model for malaria transmission dynamics incorporating seasonality and awareness campaign.}, journal = {Infectious Disease Modelling}, volume = {9}, number = {1}, pages = {84-102}, pmid = {38125201}, issn = {2468-0427}, abstract = {Malaria, a devastating disease caused by the Plasmodium parasite and transmitted through the bites of female Anopheles mosquitoes, remains a significant public health concern, claiming over 600,000 lives annually, predominantly among children. Novel tools, including the application of Wolbachia, are being developed to combat malaria-transmitting mosquitoes. This study presents a modified susceptible-exposed-infectious-recovered-susceptible (SEIRS) compartmental mathematical model to evaluate the impact of awareness-based control measures on malaria transmission dynamics, incorporating mosquito interactions and seasonality. Employing the next-generation matrix approach, we calculated a basic reproduction number (R0) of 2.4537, indicating that without robust control measures, the disease will persist in the human population. The model equations were solved numerically using fourth and fifth-order Runge-Kutta methods. The model was fitted to malaria incidence data from Kenya spanning 2000 to 2021 using least squares curve fitting. The fitting algorithm yielded a mean absolute error (MAE) of 2.6463 when comparing the actual data points to the simulated values of infectious human population (Ih). This finding indicates that the proposed mathematical model closely aligns with the recorded malaria incidence data. The optimal values of the model parameters were estimated from the fitting algorithm, and future malaria dynamics were projected for the next decade. The research findings suggest that social media-based awareness campaigns, coupled with specific optimization control measures and effective management methods, offer the most cost-effective approach to managing malaria.}, } @article {pmid38105949, year = {2023}, author = {Shropshire, JD and Conner, WR and Vanderpool, D and Hoffmann, AA and Turelli, M and Cooper, BS}, title = {Rapid turnover of pathogen-blocking Wolbachia and their incompatibility loci.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38105949}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, abstract = {At least half of all insect species carry maternally inherited Wolbachia alphaproteobacteria, making Wolbachia the most common endosymbionts in nature. Wolbachia spread to high frequencies is often due to cytoplasmic incompatibility (CI), a Wolbachia-induced sperm modification that kills embryos without Wolbachia. Several CI-causing Wolbachia variants, including wMel from Drosophila melanogaster, also block viruses. Establishing pathogen-blocking wMel in natural Aedes aegypti mosquito populations has reduced dengue disease incidence, with one study reporting about 85% reduction when wMel frequency is high. However, wMel transinfection establishment is challenging in many environments, highlighting the importance of identifying CI-causing Wolbachia variants that stably persist in diverse hosts and habitats. We demonstrate that wMel-like variants have naturally established in widely distributed holometabolous dipteran and hymenopteran insects that diverged approximately 350 million years ago, with wMel variants spreading rapidly among these hosts over only the last 100,000 years. Wolbachia genomes contain prophages that encode CI-causing operons (cifs). These cifs move among Wolbachia genomes - with and without prophages - even more rapidly than Wolbachia move among insect hosts. Our results shed light on how rapid host switching and horizontal gene transfer contribute to Wolbachia and cif diversity in nature. The diverse wMel variants we report here from hosts present in different climates offer many new options for broadening Wolbachia-based biocontrol of diseases and pests.}, } @article {pmid38105374, year = {2023}, author = {Cain, JL and Norris, JK and Swan, MP and Nielsen, MK}, title = {A diverse microbial community and common core microbiota associated with the gonad of female Parascaris spp.}, journal = {Parasitology research}, volume = {123}, number = {1}, pages = {56}, pmid = {38105374}, issn = {1432-1955}, mesh = {Animals ; Horses ; Female ; *Ascaridoidea/genetics ; *Horse Diseases/parasitology ; *Ascaridida Infections/veterinary/parasitology ; Drug Resistance ; Feces/parasitology ; *Parasites ; *Helminths ; Gonads ; }, abstract = {The microbiome plays an important role in health, where changes in microbiota composition can have significant downstream effects within the host, and host-microbiota relationships can be exploited to affect health outcomes. Parasitic helminths affect animals globally, but an exploration of their microbiota has been limited, despite the development of anti-Wolbachia drugs to help control infections with some filarial nematodes. The equine ascarids, Parascaris spp., are considered the most pathogenic nematodes affecting juvenile horses and are also the only ascarid parasite to have developed widespread anthelmintic resistance. The aim of this study was to characterize the microbiota of this helminth, focusing on the female gonad, determine a core microbiota for this organ, identify bacterial species, and show bacterial localization to the female gonad via in situ hybridization (ISH). A total of 22 gonads were isolated from female Parascaris spp. collected from three foals, and 9 female parasites were formalin-fixed and paraffin-embedded for ISH. Next-generation sequencing was performed using V3-V4 primers as well as the Swift Amplicon™ 16S+ ITS Panel. Overall, ten genera were identified as members of the Parascaris spp. female gonad and twelve bacterial species were identified. The most prevalent genus was Mycoplasma, followed by Reyranella, and there were no differences in alpha diversity between parasites from different horses. Specific eubacteria staining was identified in both the intestine and within the gonad using ISH. Overall, this study provided in-depth information regarding the female Parascaris spp. microbiota and was the first to identify the core microbiota within a specific parasite organ.}, } @article {pmid38104431, year = {2024}, author = {Lau, DC and Power, RI and Šlapeta, J}, title = {Exploring multiplex qPCR as a diagnostic tool for detecting microfilarial DNA in dogs infected with Dirofilaria immitis: A comparative analysis with the modified Knott's test.}, journal = {Veterinary parasitology}, volume = {325}, number = {}, pages = {110097}, doi = {10.1016/j.vetpar.2023.110097}, pmid = {38104431}, issn = {1873-2550}, mesh = {Animals ; Dogs ; *Dirofilaria immitis/genetics ; Microfilariae/genetics ; Australia ; *Dog Diseases/diagnosis ; *Dirofilariasis/diagnosis ; DNA ; Lactones ; }, abstract = {Current recommendations to diagnose cardiopulmonary dirofilariosis in dogs caused by Dirofilaria immitis involves tandem antigen and circulating microfilariae tests. The modified Knott's test is an important tool in heartworm diagnosis, allowing identification of circulating microfilariae. However, the subjective nature of the modified Knott's test affects its accuracy and diagnostic laboratories usually do not provide a quantitative outcome. Quantitative enumeration of microfilariae enables clinicians to track treatment progress and acts as a proxy for detecting emerging macrocyclic lactone resistance. There is a need for better diagnostic tools suitable for routine use to efficiently and accurately quantify the presence of D. immitis microfilaremia. The aim of this study was to determine whether the quantitative modified Knott's test can be substituted by multiplex quantitative polymerase chain reaction (qPCR) targeting D. immitis and associated Wolbachia endosymbiont DNA in canine blood samples. To do this, genomic DNA samples (n = 161) from Australian dogs, collected as part of a previous 2021 study, were assessed in a TaqMan qPCR targeting DNA of D. immitis, Wolbachia sp. and Canis lupus familiaris. Of the 161 genomic DNA samples, eight were considered positive for D. immitis microfilariae. The qPCR assay demonstrated good efficiency (E = 90 to 110%, R[2] > 0.94). Considering the performance and efficient use of bench time, this TaqMan qPCR assay is a suitable alternative to the modified Knott's test for quantitative enumeration of microfilariae (Cohen's kappa coefficient [κ]: κ = 1 using D. immitis qPCR marker, κ = 0.93 using Wolbachia qPCR marker). The qPCR data demonstrated a comparable result to that of the quantitative modified Knott's test in a 2022 survey of D. immitis in Australian dogs (n = 23) before and after macrocyclic lactone (ML) administration. Improving the detection and diagnosis of canine heartworm infections will assist veterinarians in better managing and controlling disease outcomes and will be valuable for tracking the spread of ML resistance in Australia.}, } @article {pmid38100392, year = {2023}, author = {Malavige, GN and Sjö, P and Singh, K and Piedagnel, JM and Mowbray, C and Estani, S and Lim, SCL and Siquierra, AM and Ogg, GS and Fraisse, L and Ribeiro, I}, title = {Facing the escalating burden of dengue: Challenges and perspectives.}, journal = {PLOS global public health}, volume = {3}, number = {12}, pages = {e0002598}, pmid = {38100392}, issn = {2767-3375}, abstract = {Dengue is the most rapidly emerging mosquito-borne infection and, due to climate change and unplanned urbanization, it is predicted that the global burden of dengue will rise further as the infection spreads to new geographical locations. Dengue-endemic countries are often unable to cope with such increases, with health care facilities becoming overwhelmed during each dengue season. Furthermore, although dengue has been predominantly a childhood illness in the past, it currently mostly affects adults in many countries, with higher incidence of severe disease and mortality rates in pregnant women and in those with comorbidities. As there is currently no specific treatment for dengue and no early biomarker to identify those who will progress to develop vascular leakage, all individuals with dengue are closely monitored in case they need fluid management. Furthermore, diagnosing patients with acute dengue is challenging due to the similarity of clinical symptoms during early illness and poor sensitivity and specificity of point-of-care diagnostic tests. Novel vector control methods, such as the release of Wolbachia-infected mosquitoes, have shown promising results by reducing vector density and dengue incidence in clinical trial settings. A new dengue vaccine, TAK-003, had an efficacy of 61.2% against virologically confirmed dengue, 84.1% efficacy against hospitalizations and a 70% efficacy against development of dengue haemorrhagic fever (DHF) at 54 months. While vaccines and mosquito control methods are welcome, they alone are unlikely to fully reduce the burden of dengue, and a treatment for dengue is therefore essential. Several novel antiviral drugs are currently being evaluated along with drugs that inhibit host mediators, such as mast cell products. Although viral proteins such as NS1 contribute to the vascular leak observed in severe dengue, the host immune response to the viral infection also plays a significant role in progression to severe disease. There is an urgent need to discover safe and effective treatments for dengue to prevent disease progression.}, } @article {pmid38076953, year = {2023}, author = {Fricke, LC and Lindsey, AR}, title = {Identification of parthenogenesis-inducing effector proteins in Wolbachia.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38076953}, support = {R35 GM150991/GM/NIGMS NIH HHS/United States ; }, abstract = {Bacteria in the genus Wolbachia have evolved numerous strategies to manipulate arthropod sex, including the conversion of would-be male offspring to asexually reproducing females. This so-called "parthenogenesis-induction" phenotype can be found in a number of Wolbachia strains that infect arthropods with haplodiploid sex determination systems, including parasitoid wasps. Despite the discovery of microbe-mediated parthenogenesis more than 30 years ago, the underlying genetic mechanisms have remained elusive. We used a suite of genomic, computational, and molecular tools to identify and characterize two proteins that are uniquely found in parthenogenesis-inducing Wolbachia and have strong signatures of host-associated bacterial effector proteins. These putative parthenogenesis-inducing proteins have structural homology to eukaryotic protein domains including nucleoporins, the key insect sex-determining factor Transformer, and a eukaryotic-like serine-threonine kinase with leucine rich repeats. Furthermore, these proteins significantly impact eukaryotic cell biology in the model, Saccharomyces cerevisiae. We suggest these proteins are parthenogenesis-inducing factors and our results indicate this would be made possible by a novel mechanism of bacterial-host interaction.}, } @article {pmid38071646, year = {2023}, author = {Urairi, C and Fujito, S}, title = {Interbiotype hybridization between biotypes A and B of Liriomyza chinensis (Diptera: Agromyzidae).}, journal = {Journal of economic entomology}, volume = {}, number = {}, pages = {}, doi = {10.1093/jee/toad223}, pmid = {38071646}, issn = {1938-291X}, abstract = {Liriomyza chinensis (Kato) is a formidable pest of Allium species, especially the Japanese bunching onion Allium fistulosum L. Recently, a novel biotype of L. chinensis (biotype B) has emerged, which causes more severe damage than the native biotype A. It has been reported that biotype B has frequently displaced biotype A in the Japanese bunching onion fields in Japan. As interbiotype hybridization is a possible factor that influences such displacement, interbiotype hybridization was conducted between L. chinensis biotypes A and B. Eggs were not laid under one-by-one crossing conditions; however, adult hybrid progeny of both sexes emerged from no-choice mating combinations-when multiple males and females were present. The fertility of F1 hybrid adults was also investigated, and backcrossed adults emerged from F1 females in both mating combinations. F1 males might have exhibited reproductive abnormalities because only a small number of backcross progeny emerged from the mating combinations using F1 males. Additionally, 3 representative endosymbionts (Wolbachia, Spiroplasma, and Cardinium) were investigated, and both biotypes were found to be infected by the same strain of Wolbachia. In addition, the courtship signals (tapping) of male adults differed between biotypes A and B as well as between F1 hybrids; the F1 males exhibited tapping behavior that was intermediate between biotypes A and B. Therefore, mating sounds serve as a form of premating reproductive isolation between biotypes A and B.}, } @article {pmid38070178, year = {2023}, author = {Arai, H and Ueda, M and Hirano, T and Akizuki, N and Lin, SR and Hanh, DK and Widada, J and Rohman, MS and Nakai, M and Kunimi, Y and Vang, LV and Wijonarko, A and Inoue, MN}, title = {Conserved infections and reproductive phenotypes of Wolbachia symbionts in Asian tortrix moths.}, journal = {Environmental microbiology reports}, volume = {}, number = {}, pages = {}, doi = {10.1111/1758-2229.13219}, pmid = {38070178}, issn = {1758-2229}, support = {19J13123//Japan Society for the Promotion of Science/ ; 21J00895//Japan Society for the Promotion of Science/ ; 21KK0105//Japan Society for the Promotion of Science/ ; 1539057329//JSPS Overseas Challenge Program for Young Researchers (2019) with RISTEK Foreign Research Permit/ ; }, abstract = {Wolbachia is a ubiquitous endosymbiotic bacterium that manipulates insect reproduction. A notable feature of Wolbachia is male killing (MK), whereby sons of infected females are killed during development; however, the evolutionary processes by which Wolbachia acquired the MK ability remain unclear. The tea tortrix moth Homona magnanima (Tortricidae) harbours three non-MK Wolbachia strains (wHm-a, wHm-b and wHm-c) and an MK strain wHm-t. Although wHm-t and wHm-c are closely related, only wHm-t has an MK-associated prophage region. To understand the evolutionary processes underlying the emergence of MK wHm-t, we examined Wolbachia infections and phenotypes in 62 tortricid species collected from 39 localities across Japan, Taiwan, Vietnam and Indonesia. PCR assays detected wHm-c relatives in 51 species and triple infection of wHm-a, wHm-b and wHm-c in 31 species. Apart from Taiwanese H. magnanima, no species exhibited the MK phenotype and were positive for the wHm-t-specific prophage. While wHm-t infection was dominant in Taiwanese H. magnanima, wHm-a, wHm-b and wHm-c were dominant in Japanese H. magnanima populations. These results suggest that wHm-a, wHm-b and wHm-c strains descended from a common ancestor with repeated infection loss and that wHm-t evolved from the wHm-c acquiring MK ability in allopatric populations of H. magnanima.}, } @article {pmid38060903, year = {2023}, author = {Valdez-Delgado, KM and Ríos-Delgado, JC and Nettel-Cruz, JA and Angulo-Kladt, R and Villarreal-Treviño, C}, title = {Aerial release of Aedes aegypti male mosquitoes using an unmanned aerial vehicle: a novel control strategy.}, journal = {Salud publica de Mexico}, volume = {65}, number = {4, jul-ago}, pages = {387-393}, doi = {10.21149/14466}, pmid = {38060903}, issn = {1606-7916}, mesh = {Humans ; Animals ; Female ; Male ; *Aedes ; Unmanned Aerial Devices ; Temperature ; Sexual Behavior ; *Wolbachia ; Mosquito Control/methods ; }, abstract = {OBJECTIVE: To development of a methodology for the chilling, handling, transport, and release of male Aedes aegypti mosquitoes, reared in insectary conditions to release in the field with unmanned vehicles to compete sexually with wild males in the field.

MATERIALS AND METHODS: A population of Ae. aegypti from different areas in Tapachula, Chiapas, was used. Laboratory tests were conducted: Effect of temperature and cooling time on the knockdown, recovery of males, and copulatory success.

RESULTS: The chilling temperature of 3 ± 1ºC for 30 min, was used as a knockdown temperature before handling, packing, transportation, and aerial release. The males subjected to the entire process, including the semi-field aerial release test, showed normal sexual behavior activity, obtaining 100% of females inseminated.

CONCLUSION: These results present the feasibility of applying a new control methodology using unmanned aerial vehicle (UAV) as support for the sterile insect release technique (SIT), use of Wolbachia or both, in male Ae. aegypti, for the design of strategies to control their populations.}, } @article {pmid38060854, year = {2023}, author = {López-Ordóñez, T and Díaz-Rodarte, KI and Torres-Monzón, JA and Casas-Martínez, M and Danis-Lozano, R and Mosso-González, C}, title = {[Not Available].}, journal = {Salud publica de Mexico}, volume = {65}, number = {2 mar-abr}, pages = {136-143}, doi = {10.21149/14154}, pmid = {38060854}, issn = {1606-7916}, abstract = {OBJETIVO: Analizar la expresión diferencial de proteínas de Aedes aegypti infectados con Wolbachia y su asociación con el ciclo viral del virus dengue (DENV). Material y métodos. Se revisó una base de datos de proteínas de Ae. aegypti infec-tados y no infectados con Wolbachia, cepa wMel y se buscaron estas en revistas indizadas, que hablaran de la proteína y el ciclo viral de DENV.

RESULTADOS: La expresión diferencial de proteínas de los mosquitos durante la infección con Wolbachia intervienen en los procesos de entrada, replicación y salida del DENV.

CONCLUSIONES: Existen cambios en la expresión de proteínas de células infectadas con Wolbachia, que son necesarias para el ciclo de replicación de DENV, explicando porque algunos mosquitos infectados con Wolbachia son refractarios a la infección por DENV.}, } @article {pmid38052643, year = {2023}, author = {Liu, Z and Zhou, T}, title = {The effect of migration on transmission of Wolbachia in Nilaparvata lugens.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {20}, number = {11}, pages = {20213-20244}, doi = {10.3934/mbe.2023895}, pmid = {38052643}, issn = {1551-0018}, mesh = {Animals ; *Wolbachia ; *Hemiptera ; Insecta ; *Oryza ; }, abstract = {Brown planthopper Nilaparvata lugens, which can transmit rice ragged stunt virus, is a serious and damaging pest to rice plants. Rice plants can protect themselves from the associated diseases of N.lugens by either suppressing or replacing N.lugens by releasing N.lugens infected by a special strain of Wolbachia wStri. The long-distance migration habit of N.lugens is one of the important precursors leading up to the large-scale occurrence of N.lugens. To study the effect of migration on the transmission of Wolbachia in N.lugens, a Wolbachia spreading dynamics model with migration of N.lugens between two patches is put forward. The existence and local stability conditions of equilibrium points of the system and its subsystems are obtained. Moreover, the effects of migration on the dynamic properties and the control of N.lugens are analyzed; the results show that the system can exhibit a bistable phenomenon, and the migration can change the stability of equilibrium infected with wStri from stable to unstable. The quantitative control methods for the migration of the insect N.lugens are proposed, which provide a theoretical guidance for future field experiments. Lastly, we use the Markov chain Monte Carlo (MCMC) method to estimate the parameters of the wild N.lugens migration model based on limited observational data; the numerical simulation results show that migration can increase the quantity of N.lugens, which is consistent with the relevant experimental results.}, } @article {pmid38047691, year = {2024}, author = {Nakabachi, A and Suzaki, T}, title = {Ultrastructure of the bacteriome and bacterial symbionts in the Asian citrus psyllid, Diaphorina citri.}, journal = {Microbiology spectrum}, volume = {12}, number = {1}, pages = {e0224923}, pmid = {38047691}, issn = {2165-0497}, support = {21687020, 26292174, 20H02998//MEXT | Japan Society for the Promotion of Science (JSPS)/ ; }, mesh = {Animals ; *Hemiptera ; Bacteria/genetics ; Symbiosis ; }, abstract = {Omics analyses suggested a mutually indispensable tripartite association among the host D. citri and organelle-like bacteriome associates, Carsonella and Profftella, which are vertically transmitted through host generations. This relationship is based on the metabolic complementarity among these organisms, which is partly enabled by horizontal gene transfer between partners. However, little was known about the fine morphology of the symbionts and the bacteriome, the interface among these organisms. As a first step to address this issue, the present study performed transmission electron microscopy, which revealed previously unrecognized ultrastructures, including aggregations of ribosomes in Carsonella, numerous tubes and occasional protrusions of Profftella, apparently degrading Profftella, and host organelles with different abundance and morphology in distinct cell types. These findings provide insights into the behaviors of the symbionts and host cells to maintain the symbiotic relationship in D. citri.}, } @article {pmid38037192, year = {2023}, author = {Lühken, R and Brattig, N and Becker, N}, title = {Introduction of invasive mosquito species into Europe and prospects for arbovirus transmission and vector control in an era of globalization.}, journal = {Infectious diseases of poverty}, volume = {12}, number = {1}, pages = {109}, pmid = {38037192}, issn = {2049-9957}, support = {01Kl2022//Bundesministerium für Bildung und Forschung/ ; }, mesh = {Animals ; Humans ; *Arboviruses ; Introduced Species ; Europe/epidemiology ; Mosquito Vectors ; *Aedes ; *Zika Virus ; *Zika Virus Infection ; Mosquito Control ; }, abstract = {BACKGROUND: Mosquito research in Europe has a long history, primarily focused on malaria vectors. In recent years, invasive mosquito species like the Asian tiger mosquito (Aedes albopictus) and the spread of arboviruses like dengue virus, chikungunya virus or bluetongue virus have led to an intensification of research and monitoring in Europe. The risk of further dissemination of exotic species and mosquito-borne pathogens is expected to increase with ongoing globalization, human mobility, transport geography, and climate warming. Researchers have conducted various studies to understand the ecology, biology, and effective control strategies of mosquitoes and associated pathogens.

MAIN BODY: Three invasive mosquito species are established in Europe: Asian tiger mosquito (Aedes albopictus), Japanese bush mosquito (Ae. japonicus), and Korean bush mosquito (Aedes koreicus). Ae. albopictus is the most invasive species and has been established in Europe since 1990. Over the past two decades, there has been an increasing number of outbreaks of infections by mosquito-borne viruses in particular chikungunya virus, dengue virus or Zika virus in Europe primary driven by Ae. albopictus. At the same time, climate change with rising temperatures results in increasing threat of invasive mosquito-borne viruses, in particular Usutu virus and West Nile virus transmitted by native Culex mosquito species. Effective mosquito control programs require a high level of community participation, going along with comprehensive information campaigns, to ensure source reduction and successful control. Control strategies for container breeding mosquitoes like Ae. albopictus or Culex species involve community participation, door-to-door control activities in private areas. Further measures can involve integration of sterile insect techniques, applying indigenous copepods, Wolbachia sp. bacteria, or genetically modified mosquitoes, which is very unlike to be practiced as standard method in the near future.

CONCLUSIONS: Climate change and globalization resulting in the increased establishment of invasive mosquitoes in particular of the Asian tiger mosquito Ae. albopictus in Europe within the last 30 years and increasing outbreaks of infections by mosquito-borne viruses warrants intensification of research and monitoring. Further, effective future mosquito control programs require increase in intense community and private participation, applying physical, chemical, biological, and genetical control activities.}, } @article {pmid38036023, year = {2024}, author = {Ngnindji-Youdje, Y and Lontsi-Demano, M and Diarra, AZ and Makaila, AM and Tchuinkam, T and Berenger, JM and Parola, P}, title = {Morphological, molecular, and MALDI-TOF MS identification of bed bugs and associated Wolbachia species from Cameroon.}, journal = {Acta tropica}, volume = {249}, number = {}, pages = {107086}, doi = {10.1016/j.actatropica.2023.107086}, pmid = {38036023}, issn = {1873-6254}, mesh = {Male ; Animals ; Female ; *Bedbugs/genetics/microbiology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; *Wolbachia/genetics ; Cameroon ; *Ectoparasitic Infestations ; }, abstract = {After vanishing from the public eye for more than 50 years, bed bugs have resurged to become one of the most widely discussed and heavily researched insect pests in the world. This study presents the basic information of infestations of tropical bed bugs, Cimex hemipterus (Hemiptera: Cimicidae), in Cameroon. A total of 248 immature stage and adult bed bug specimens were collected from households and a travel agency in Yaoundé and Douala, Cameroon. The ability of MALDI-TOF MS to identify bed bugs was tested using heads for adults and cephalothoraxes for immature stages. Microorganism screening was performed by qPCR and confirmed by regular PCR and sequencing. Based on morphometrical criteria, four stages of immature bed bugs are represented. Of the 248 bed bug specimens morphologically identified as Cimex hemipterus, 246 (77 males, 65 females and 104 immature specimens) were submitted to MALDI-TOF MS analysis. Of the 222 adults and immature specimens tested, 122 (59.9 %) produced good quality MALDI-TOF MS spectra (35 adults and 87 immature specimens). Blind testing allowed species level identification of 98.21 % of adult and immature C. hemipterus. Among the bacteria tested, only Wolbachia DNA was found in 12/246 (4.8 %) bed bugs. More surveys in the country are warranted to assess the true level of bed bug infestations, in order to take appropriate action for their control.}, } @article {pmid38032857, year = {2023}, author = {Velez, ID and Tanamas, SK and Arbelaez, MP and Kutcher, SC and Duque, SL and Uribe, A and Zuluaga, L and Martínez, L and Patiño, AC and Barajas, J and Muñoz, E and Mejia Torres, MC and Uribe, S and Porras, S and Almanza, R and Pulido, H and O'Neill, SL and Santacruz-Sanmartin, E and Gonzalez, S and Ryan, PA and Denton, JA and Jewell, NP and Dufault, SM and Simmons, CP and Anders, KL}, title = {Reduced dengue incidence following city-wide wMel Wolbachia mosquito releases throughout three Colombian cities: Interrupted time series analysis and a prospective case-control study.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {11}, pages = {e0011713}, pmid = {38032857}, issn = {1935-2735}, mesh = {Animals ; Humans ; *Wolbachia ; Colombia/epidemiology ; *Dengue Virus ; Cities/epidemiology ; Incidence ; Interrupted Time Series Analysis ; Case-Control Studies ; Reproducibility of Results ; Pest Control, Biological/methods ; *Aedes ; *Dengue/epidemiology/prevention & control ; Mosquito Vectors ; }, abstract = {BACKGROUND: The introduction of Wolbachia (wMel strain) into Aedes aegypti mosquitoes reduces their capacity to transmit dengue and other arboviruses. Randomised and non-randomised studies in multiple countries have shown significant reductions in dengue incidence following field releases of wMel-infected Ae. aegypti. We report the public health outcomes from phased, large-scale releases of wMel-Ae. aegypti mosquitoes throughout three contiguous cities in the Aburrá Valley, Colombia.

Following pilot releases in 2015-2016, staged city-wide wMel-Ae. aegypti deployments were undertaken in the cities of Bello, Medellín and Itagüí (3.3 million people) between October 2016 and April 2022. The impact of the Wolbachia intervention on dengue incidence was evaluated in two parallel studies. A quasi-experimental study using interrupted time series analysis showed notified dengue case incidence was reduced by 95% in Bello and Medellín and 97% in Itagüí, following establishment of wMel at ≥60% prevalence, compared to the pre-intervention period and after adjusting for seasonal trends. A concurrent clinic-based case-control study with a test-negative design was unable to attain the target sample size of 63 enrolled virologically-confirmed dengue (VCD) cases between May 2019 and December 2021, consistent with low dengue incidence throughout the Aburrá Valley following wMel deployments. Nevertheless, VCD incidence was 45% lower (OR 0.55 [95% CI 0.25, 1.17]) and combined VCD/presumptive dengue incidence was 47% lower (OR 0.53 [95% CI 0.30, 0.93]) among participants resident in wMel-treated versus untreated neighbourhoods.

CONCLUSIONS/SIGNIFICANCE: Stable introduction of wMel into local Ae. aegypti populations was associated with a significant and sustained reduction in dengue incidence across three Colombian cities. These results from the largest contiguous Wolbachia releases to-date demonstrate the real-world effectiveness of the method across large urban populations and, alongside previously published results, support the reproducibility of this effectiveness across different ecological settings.

TRIAL REGISTRATION: NCT03631719.}, } @article {pmid38032856, year = {2023}, author = {Velez, ID and Uribe, A and Barajas, J and Uribe, S and Ángel, S and Suaza-Vasco, JD and Mejia Torres, MC and Arbeláez, MP and Santacruz-Sanmartin, E and Duque, L and Martínez, L and Posada, T and Patiño, AC and Gonzalez, SM and Velez, AL and Ramírez, J and Salazar, M and Gómez, S and Osorio, JE and Iturbe-Ormaetxe, I and Dong, Y and Muzzi, FC and Rances, E and Johnson, PH and Smithyman, R and Col, B and Green, BR and Frossard, T and Brown-Kenyon, J and Joubert, DA and Grisales, N and Ritchie, SA and Denton, JA and Gilles, JRL and Anders, KL and Kutcher, SC and Ryan, PA and O'Neill, SL}, title = {Large-scale releases and establishment of wMel Wolbachia in Aedes aegypti mosquitoes throughout the Cities of Bello, Medellín and Itagüí, Colombia.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {11}, pages = {e0011642}, pmid = {38032856}, issn = {1935-2735}, mesh = {Animals ; Humans ; *Aedes ; *Wolbachia ; Cities ; Colombia ; Environment ; Mosquito Vectors ; }, abstract = {BACKGROUND: The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and has been shown to reduce the transmission of dengue and other Aedes-borne viruses. Here we report the entomological results from phased, large-scale releases of Wolbachia infected Ae. aegypti mosquitoes throughout three contiguous cities located in the Aburrá Valley, Colombia.

Local wMel Wolbachia-infected Ae. aegypti mosquitoes were generated and then released in an initial release pilot area in 2015-2016, which resulted in the establishment of Wolbachia in the local mosquito populations. Subsequent large-scale releases, mainly involving vehicle-based releases of adult mosquitoes along publicly accessible roads and streets, were undertaken across 29 comunas throughout Bello, Medellín and Itagüí Colombia between 2017-2022. In 9 comunas these were supplemented by egg releases that were undertaken by staff or community members. By the most recent monitoring, Wolbachia was found to be stable and established at consistent levels in local mosquito populations (>60% prevalence) in the majority (67%) of areas.

CONCLUSION: These results, from the largest contiguous releases of wMel Wolbachia mosquitoes to date, highlight the operational feasibility of implementing the method in large urban settings. Based on results from previous studies, we expect that Wolbachia establishment will be sustained long term. Ongoing monitoring will confirm Wolbachia persistence in local mosquito populations and track its establishment in the remaining areas.}, } @article {pmid38021190, year = {2023}, author = {Bawm, S and Khaing, Y and Chel, HM and Hmoon, MM and Win, SY and Bo, M and Naing, T and Htun, LL}, title = {Molecular detection of Dirofilaria immitis and its Wolbachia endosymbionts in dogs from Myanmar.}, journal = {Current research in parasitology & vector-borne diseases}, volume = {4}, number = {}, pages = {100148}, pmid = {38021190}, issn = {2667-114X}, abstract = {Heartworm disease in dogs and cats caused by Dirofilaria immitis continues to be a major clinical issue globally. This study focused on dogs suspicious of having tick-borne diseases (TBD) brought to a clinic and a veterinary teaching hospital in Myanmar. Blood samples were collected and initially screened using SNAP® 4Dx® Plus test kit. All dog blood samples were subjected to conventional PCR to detect both Dirofilaria spp. (cox1 gene) and Wolbachia spp. (16S rDNA) infections. Infection with D. immitis was detected in 14 (28.0%) of 50 examined samples, while the detection rate of TBD causative agents, including Anaplasma phagocytophilum and Ehrlichia canis, was 26.0% (13/50) and 26.0% (13/50), respectively, as determined by ELISA rapid test. In this study, D. immitis infection was moderately but significantly correlated with TBD infections (Pearson's r = 0.397, P = 0.008). Comparative sequence and phylogenetic analyses provided molecular identification of D. immitis in Myanmar and confirmed the identity of its Wolbachia endosymbiont with Wolbachia endosymbionts isolated from D. immitis, Rhipicephalus sanguineus and Aedes aegypti. The present study contributes to our understanding of the coexistence of D. immitis and Wolbachia endosymbiosis in dogs, and the findings may benefit the future prevention and control of dirofilariasis in dogs.}, } @article {pmid38020682, year = {2023}, author = {McPherson, AE and Abram, PK and Curtis, CI and Wannop, ER and Dudzic, JP and Perlman, SJ}, title = {Dynamic changes in Wolbachia infection over a single generation of Drosophila suzukii, across a wide range of resource availability.}, journal = {Ecology and evolution}, volume = {13}, number = {11}, pages = {e10722}, pmid = {38020682}, issn = {2045-7758}, abstract = {Wolbachia bacteria are maternally inherited symbionts that commonly infect terrestrial arthropods. Many Wolbachia reach high frequencies in their hosts by manipulating their reproduction, for example by causing reproductive incompatibilities between infected male and uninfected female hosts. However, not all strains manipulate reproduction, and a key unresolved question is how these non-manipulative Wolbachia persist in their hosts, often at intermediate to high frequencies. One such strain, wSuz, infects the invasive fruit pest Drosophila suzukii, spotted-wing drosophila. Here, we tested the hypothesis that wSuz infection provides a competitive benefit when resources are limited. Over the course of one season, we established population cages with varying amounts of food in a semi-field setting and seeded them with a 50:50 mixture of flies with and without Wolbachia. We predicted that Wolbachia-infected individuals should have higher survival and faster development than their uninfected counterparts when there was little available food. We found that while food availability strongly impacted fly fitness, there was no difference in development times or survival between Wolbachia-infected and uninfected flies. Interestingly, however, Wolbachia infection frequencies changed dramatically, with infections either increasing or decreasing by as much as 30% in a single generation, suggesting the possibility of unidentified factors shaping Wolbachia infection over the course of the season.}, } @article {pmid38018822, year = {2023}, author = {Dai, X and Quan, Q and Jiao, J}, title = {Modelling and analysis of periodic impulsive releases of the Nilaparvata lugens infected with wStri-Wolbachia.}, journal = {Journal of biological dynamics}, volume = {17}, number = {1}, pages = {2287077}, doi = {10.1080/17513758.2023.2287077}, pmid = {38018822}, issn = {1751-3766}, mesh = {Animals ; *Wolbachia ; Models, Biological ; Population Dynamics ; *Hemiptera ; }, abstract = {In this paper, we formulate a population suppression model and a population replacement model with periodic impulsive releases of Nilaparvata lugens infected with wStri. The conditions for the stability of wild-N.lugens-eradication periodic solution of two systems are obtained by applying the Floquet theorem and comparison theorem. And the sufficient conditions for the persistence in the mean of wild N.lugens are also given. In addition, the sufficient conditions for the extinction and persistence of the wild N.lugens in the subsystem without wLug are also obtained. Finally, we give numerical analysis which shows that increasing the release amount or decreasing the release period are beneficial for controlling the wild N.lugens, and the efficiency of population replacement strategy in controlling wild populations is higher than that of population suppression strategy under the same release conditions.}, } @article {pmid38016137, year = {2023}, author = {Azarm, A and Koosha, M and Dalimi, A and Zahraie-Ramazani, A and Akhavan, AA and Saeidi, Z and Mohebali, M and Azam, K and Vatandoost, H and Oshaghi, MA}, title = {Association Between Wolbachia Infection and Susceptibility to Deltamethrin Insecticide in Phlebotomus papatasi (Diptera: Psychodidae), the Main Vector of Zoonotic Cutaneous Leishmaniasis.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {}, number = {}, pages = {}, doi = {10.1089/vbz.2023.0085}, pmid = {38016137}, issn = {1557-7759}, abstract = {Background: Phlebotomus papatasi (Diptera: Psychodidae) is the main vector of zoonotic cutaneous leishmaniasis. Wolbachia is a symbiotic alphaproteobacteria of arthropods that can be involved in susceptibility or resistance. This study aimed to investigate the relationship between Wolbachia and Deltamethrin susceptibility/resistance in Ph. papatasi. Deltamethrin filter papers (0.00002%) were used to test sand fly field collected from southern Iran. After the test, PCR amplification of the Wolbachia surface protein gene (wsp) was used to measure Wolbachia infection rate in the killed, surviving, and control groups. Result: The rates of infection by Wolbachia strain (wPap, super group A) differed between killed (susceptible) and surviving (resistant) Ph. papatasi specimens. The rate of Wolbachia infection in susceptible individuals was more than twice (2.3) (39% vs. 17%) in resistant individuals with the same genetic background. This difference was highly significant (p < 0.001), indicating a positive association between Wolbachia infection and susceptibility to Deltamethrin. In addition, the results showed that Deltamethrin can act as a PCR inhibitor during detection of Wolbachia in Ph. papatasi. Conclusion: Results of this study show that Wolbachia is associated with Deltamethrin susceptibility level in Ph. papatasi. Also, as Deltamethrin has been identified as a PCR inhibitor, great care must be taken in interpreting Wolbachia infection status in infected populations. The results of this study may provide information for a better understanding of the host-symbiont relationship, as well as application of host symbiosis in pest management.}, } @article {pmid38012718, year = {2023}, author = {Zhang, H and Trueman, E and Hou, X and Chew, X and Deng, L and Liew, J and Chia, T and Xi, Z and Tan, CH and Cai, Y}, title = {Different mechanisms of X-ray irradiation-induced male and female sterility in Aedes aegypti.}, journal = {BMC biology}, volume = {21}, number = {1}, pages = {274}, pmid = {38012718}, issn = {1741-7007}, mesh = {Humans ; Animals ; Male ; Female ; *Infertility, Female ; X-Rays ; *Aedes ; Mosquito Vectors ; Mosquito Control/methods ; *Wolbachia ; Insecta ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {BACKGROUND: Aedes aegypti (Ae. aegypti) is the major vector that transmits many diseases including dengue, Zika, and filariasis in tropical and subtropical regions. Due to the growing resistance to chemical-based insecticides, biological control methods have become an emerging direction to control mosquito populations. The sterile insect technique (SIT) deploys high doses of ionizing radiation to sterilize male mosquitoes before the release. The Wolbachia-based population suppression method of the incompatible insect technique (IIT) involves the release of Wolbachia-infected males to sterilize uninfected field females. Due to the lack of perfect sex separation tools, a low percentage of female contamination is detected in the male population. To prevent the unintentional release of these Wolbachia-infected females which might result in population replacement, a low dose of X-ray irradiation is deployed to sterilize any female escapees. However, it remains unclear whether these irradiation-induced male and female sterilizations share common mechanisms.

RESULTS: In this work, we set out to define the minimum dose of X-ray radiation required for complete female sterilization in Ae. aegypti (NEA-EHI strain). Further results showed that this minimum dose of X-ray irradiation for female sterilization significantly reduced male fertility. Similar results have been reported previously in several operational trials. By addressing the underlying causes of the sterility, our results showed that male sterility is likely due to chromosomal damage in the germ cells induced by irradiation. In contrast, female sterility appears to differ and is likely initiated by the elimination of the somatic supporting cells, which results in the blockage of the ovariole maturation. Building upon these findings, we identified the minimum dose of X-ray irradiation on the Wolbachia-infected NEA-EHI (wAlbB-SG) strain, which is currently being used in the IIT-SIT field trial. Compared to the uninfected parental strain, a lower irradiation dose could fully sterilize wAlbB-SG females. This suggests that Wolbachia-carrying mosquitoes are more sensitive to irradiation, consistent with a previous report showing that a lower irradiation dose fully sterilized Wolbachia-infected Ae. aegypti females (Brazil and Mexican strains) compared to those uninfected controls.

CONCLUSIONS: Our findings thus reveal the distinct mechanisms of ionizing X-ray irradiation-induced male or female sterility in Ae. aegypti mosquitoes, which may help the design of X-ray irradiation-based vector control methods.}, } @article {pmid38011160, year = {2023}, author = {Oliveira, TMP and Rojas, MVR and Amorim, JA and Alonso, DP and de Carvalho, DP and Ribeiro, KAN and Sallum, MAM}, title = {Bacterial diversity on larval and female Mansonia spp. from different localities of Porto Velho, Rondonia, Brazil.}, journal = {PloS one}, volume = {18}, number = {11}, pages = {e0293946}, pmid = {38011160}, issn = {1932-6203}, mesh = {Female ; Animals ; Larva/microbiology ; Brazil ; RNA, Ribosomal, 16S/genetics ; *Culicidae/genetics ; Bacteria/genetics ; }, abstract = {Studies based on the bacterial diversity present in Mansonia spp. are limited; therefore, the aim of this study was to investigate the bacterial diversity in females and larvae of Mansonia spp., describe the differences between the groups identified, and compare the microbiota of larvae from different collection sites. Sequences of the 16S rRNA region from the larvae and females of Mansonia spp. were analyzed. Diversity analyzes were performed to verify the possible bacterial differences between the groups and the collection sites. The results showed Pseudomonas was the most abundant genus in both females and larvae, followed by Wolbachia in females and Rikenellaceae and Desulfovibrio in larvae. Desulfovibrio and Sulfurospirillum, sulfate- and sulfur-reducing bacteria, respectively, were abundant on the larvae. Aminomonas, an amino acid-degrading bacterium, was found only in larvae, whereas Rickettsia was identified in females. Bacterial diversity was observed between females and larvae of Mansonia and between larvae from different collection sites. In addition, the results suggest that the environment influenced bacterial diversity.}, } @article {pmid38006562, year = {2024}, author = {Serbus, LR}, title = {A Light in the Dark: Uncovering Wolbachia-Host Interactions Using Fluorescence Imaging.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {349-373}, pmid = {38006562}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Arthropods/microbiology ; *Nematoda ; Symbiosis ; Optical Imaging ; }, abstract = {The success of microbial endosymbionts, which reside naturally within a eukaryotic "host" organism, requires effective microbial interaction with, and manipulation of, the host cells. Fluorescence microscopy has played a key role in elucidating the molecular mechanisms of endosymbiosis. For 30 years, fluorescence analyses have been a cornerstone in studies of endosymbiotic Wolbachia bacteria, focused on host colonization, maternal transmission, reproductive parasitism, horizontal gene transfer, viral suppression, and metabolic interactions in arthropods and nematodes. Fluorescence-based studies stand to continue informing Wolbachia-host interactions in increasingly detailed and innovative ways.}, } @article {pmid38006561, year = {2024}, author = {Bordenstein, SR}, title = {Isolation of Phage WO Particles from Wolbachia-Infected Arthropods.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {337-348}, pmid = {38006561}, issn = {1940-6029}, mesh = {Animals ; *Arthropods ; *Wolbachia/genetics ; *Bacteriophages/genetics ; Prophages ; Myoviridae ; }, abstract = {Nearly all arthropod-associated Wolbachia contain intact and/or genomic remnants of phage WO, temperate bacteriophages that facilitate horizontal gene transfer, genomic rearrangement of the bacterial chromosome, and symbiotic interactions between Wolbachia and their arthropod hosts. Integrated prophage WO genomes produce active, lytic particles; but the lack of a cell-free culturing system for Wolbachia render them difficult to purify and study. This chapter describes polyethylene glycol (PEG) precipitation of phage particles from Wolbachia-infected arthropods, followed by confirmation of phage WO isolation and purification using electron microscopy and PCR.}, } @article {pmid38006560, year = {2024}, author = {Murphy, RO and Beckmann, JF}, title = {Using Baker's Yeast to Determine Functions of Novel Wolbachia (and Other Prokaryotic) Effectors.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {321-336}, pmid = {38006560}, issn = {1940-6029}, mesh = {Animals ; Humans ; *Saccharomyces cerevisiae/genetics/metabolism ; *Wolbachia/genetics ; Genome ; Protein Interaction Mapping ; Insecta/genetics ; Symbiosis ; }, abstract = {Yeasts are single-celled eukaryotic organisms classified as fungi, mostly in the phylum Ascomycota. Of about 1500 named species, Saccharomyces cerevisiae, also known as baker's yeast, domesticated by humans in the context of cooking and brewing, is a profound genetic tool for exploring functions of novel effector proteins from Wolbachia and prokaryotes in general. Wolbachia is a Gram-negative alpha-proteobacterium that infects up to ~75% of all insects as an obligate intracellular microbe (Jeyaprakash A, Hoy MA, Insect Mol Biol 9:393-405, 2000). Wolbachia's lifestyle presents unique challenges for researchers. Wolbachia cannot be axenically cultured and has never been genetically manipulated. Furthermore, many Wolbachia genes have no known function or well-annotated orthologs in other genomes. Yet given the effects of Wolbachia on host phenotypes, which have considerable practical applications for pest control, they undoubtedly involve secreted effector proteins that interact with host gene products. Studying these effectors is challenging with Wolbachia's current genetic limitations. However, some of the constraints to working with Wolbachia can be overcome by expressing candidate proteins in S. cerevisiae. This approach capitalizes on yeast's small genome (~6500 genes), typical eukaryotic cellular organization, and the sophisticated suite of genetic tools available for its manipulation in culture. Thus, yeast can serve as a powerful mock eukaryotic host background to study Wolbachia effector function. Specifically, yeast is used for recombinant protein expression, drug discovery, protein localization studies, protein interaction mapping (yeast two-hybrid system), modeling chromosomal evolution, and examining interactions between proteins responsible for complex phenotypes in less tractable prokaryotic systems. As an example, the paired genes responsible for Wolbachia-mediated cytoplasmic incompatibility (CI) encode novel proteins with limited homology to other known proteins, and no obvious function. This article details how S. cerevisiae was used as an initial staging ground to explore the molecular basis of one of Wolbachia's trademark phenotypes (CI).}, } @article {pmid38006559, year = {2024}, author = {Cortez, CT and Murphy, RO and Owens, IM and Beckmann, JF}, title = {Use of Drosophila Transgenics to Identify Functions for Symbiont Effectors.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {301-320}, doi = {10.1007/978-1-0716-3553-7_18}, pmid = {38006559}, issn = {1940-6029}, mesh = {Animals ; *Drosophila ; Drosophila melanogaster/genetics ; Animals, Genetically Modified ; Biological Evolution ; Insecta ; *Wolbachia/genetics ; Symbiosis/genetics ; }, abstract = {Wolbachia, one of the most successful and studied insect symbionts, and Drosophila, one of the most understood model insects, can be exploited as complementary tools to unravel mechanisms of insect symbiosis. Although Wolbachia itself cannot be grown axenically as clonal isolates or genetically manipulated by standard methods, its reproductive phenotypes, including cytoplasmic incompatibility (CI), have been elucidated using well-developed molecular tools and precise transgenic manipulations available for Drosophila melanogaster. Current research only scratches the surface of how Drosophila can provide a tool for understanding Wolbachia's evolutionary success and the molecular roles of its genetic elements. Here, we briefly outline basic methodologies inherent to transgenic Drosophila systems that have already contributed significant advances in understanding CI, but may be unfamiliar to those who lack experience in Drosophila genetics. In the future, these approaches will continue providing significant insights into Wolbachia that undoubtedly will be extended to other insect symbionts and their biological capabilities.}, } @article {pmid38006558, year = {2024}, author = {Valerio, F and Twort, VG and Duplouy, A}, title = {A Worked Example of Screening Genomic Material for the Presence of Wolbachia Infection.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {275-299}, doi = {10.1007/978-1-0716-3553-7_17}, pmid = {38006558}, issn = {1940-6029}, mesh = {*Wolbachia/genetics ; Phylogeny ; Symbiosis/genetics ; Genomics ; Genome, Bacterial ; }, abstract = {This chapter gives a brief overview of how to screen existing host genomic data for the presence of endosymbionts, such as Wolbachia. The various programs used provide test examples, and the corresponding manuals and discussion boards provide invaluable information. Please do consult these resources.}, } @article {pmid38006557, year = {2024}, author = {Valerio, F and Twort, VG and Duplouy, A}, title = {Screening Host Genomic Data for Wolbachia Infections.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {251-274}, pmid = {38006557}, issn = {1940-6029}, mesh = {*Wolbachia/genetics ; Genome, Bacterial ; Genomics ; Base Sequence ; DNA, Bacterial/genetics ; Symbiosis/genetics ; }, abstract = {Less than a decade ago, the production of Wolbachia genomic assemblies was tedious, time-consuming, and expensive. The production of Wolbachia genomic DNA free of contamination from host DNA, as required for Wolbachia-targeted sequencing, was then only possible after the amplification and extraction of a large amount of clonal Wolbachia DNA. However, as an endosymbiotic bacterium, Wolbachia does not grow outside the host cell environment, and large-scale recovery of the bacteria required mass rearing of their host, preferably clones of a single individual to avoid strain genetic diversity, or amplification of cell cultures infected with a single Wolbachia strain. Bacterial DNA could be separated from host DNA based on genomic size. Nowadays, the production of full Wolbachia genomes does not require the physical isolation of the bacterial strains from their respective hosts, and the bacterium is often sequenced as a by-catch of host genomic projects. Here, we provide a step-by-step protocol to (1) identify whether host genome projects contain reads from associated Wolbachia and (2) isolate/retrieve the Wolbachia reads from the rest of the sequenced material. We hope this simple protocol will support many projects aiming at studying diverse Wolbachia genome assemblies.}, } @article {pmid38006555, year = {2024}, author = {Chrostek, E}, title = {Procedures for the Detection of Wolbachia-Conferred Antiviral Protection in Drosophila melanogaster.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {219-237}, pmid = {38006555}, issn = {1940-6029}, mesh = {Animals ; Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Host-Pathogen Interactions/genetics ; *Virus Diseases ; Antiviral Agents ; Symbiosis ; }, abstract = {Spread of Wolbachia infections in host populations may be enhanced by Wolbachia-conferred protection from viral pathogens. Wolbachia-infected Drosophila melanogaster survive the pathogenic effects of positive-sense single-stranded RNA virus infections at a higher rate than the flies without Wolbachia. The protection can occur with or without detectable reduction in virus titer. For the comparisons to be meaningful, Wolbachia-harboring and Wolbachia-free insects need to be genetically matched, and original populations of gut microbiota need to be restored after the removal of Wolbachia using antibiotics. Here, I describe the procedures needed to detect Wolbachia-conferred antiviral protection against Drosophila C virus measured as the difference in survival and viral titer between flies with and without Wolbachia.}, } @article {pmid38006554, year = {2024}, author = {Walker, T}, title = {Detection of Natural Wolbachia Strains in Anopheles Mosquitoes.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {205-218}, pmid = {38006554}, issn = {1940-6029}, mesh = {Animals ; Humans ; *Anopheles/genetics ; *Wolbachia/genetics ; In Situ Hybridization, Fluorescence ; Mosquito Vectors ; *Malaria ; }, abstract = {Wolbachia is an endosymbiotic bacterium that naturally infects many insect species, including mosquitoes that transmit human diseases. Wolbachia strains have been shown to inhibit the transmission of both arboviruses and malaria Plasmodium parasites. The existence of natural strains in wild Anopheles (An.) mosquitoes, the vectors of malaria parasites, in an endosymbiotic relationship is still to be fully determined. Although Wolbachia has been reported to be present in wild populations of the An. gambiae complex, the primary vectors of malaria in Sub-Saharan Africa, Wolbachia DNA sequence density and infection frequencies are low. As most studies have used highly sensitive nested PCR as the only detection method, more robust evidence is required to determine whether Wolbachia strains are established as endosymbionts in Anopheles species. Techniques such as fluorescent in situ hybridization, microbiome sequencing, and Wolbachia whole genome sequencing have provided concrete evidence for genuine Wolbachia strains in two mosquito species: An. moucheti and An. demeilloni. In this chapter, the current methodology used to determine if resident strains exist in Anopheles mosquitoes will be reviewed, including both PCR- and non-PCR-based protocols.}, } @article {pmid38006553, year = {2024}, author = {Ross, PA}, title = {Measuring Host Fitness Effects and Transmission of Wolbachia Strains in Aedes aegypti Mosquitoes.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {189-203}, pmid = {38006553}, issn = {1940-6029}, mesh = {Animals ; Male ; *Aedes ; *Wolbachia ; Mosquito Vectors ; *Arboviruses ; }, abstract = {Lines of Aedes aegypti mosquitoes infected with heritable Wolbachia bacteria are being developed and released for arbovirus control. Coordinated releases of lab-reared Wolbachia mosquitoes have reduced local disease incidence by spreading virus-blocking Wolbachia strains and by crashing mosquito populations through incompatible male releases. The phenotypic effects of Wolbachia are diverse and depend on both genetics and the environment. Accurate assessments of Wolbachia effects in mosquitoes are essential, as such effects can make the difference between success and failure of a Wolbachia release program. This chapter provides guidelines for testing key Wolbachia host effects and transmission in Aedes aegypti: the most important arbovirus vector and the most common target of Wolbachia release programs. The protocols should be useful for evaluating mosquito strains prior to field release.}, } @article {pmid38006552, year = {2024}, author = {Zhang, M and Xi, Z}, title = {Wolbachia Transinfection Via Embryonic Microinjection.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {175-188}, pmid = {38006552}, issn = {1940-6029}, mesh = {Animals ; Mosquito Vectors ; *Wolbachia ; Microinjections ; Cytoplasm ; Cytosol ; *Aedes ; Insecta ; }, abstract = {The process of transferring Wolbachia from one species to another to establish a stable, maternally inherited infection in the target species is known as transinfection. The success of transinfection is primarily achieved through embryonic microinjection, which is the most direct and efficient means of delivering Wolbachia into the germline of the target species and establishing stable maternal transmission. For the fundamental studies, transinfection is often used to characterize Wolbachia-host interactions, including Wolbachia host range, the role of host or bacterial factors in symbiosis, and evolution of Wolbachia-host associations. For the applied studies, use of transinfection to generate a novel infection in the target species is the first step to build the weapon for both population replacement and population suppression for controlling insect pests or their transmitted diseases. For the primary dengue vector Aedes aegypti and Anopheles vectors of malaria, which either do not naturally carry Wolbachia or are infected with strains that lack necessary features for implementation, transinfection can be established by introducing a novel strain capable of inducing both cytoplasmic incompatibility (CI) and pathogen blocking. For A. albopictus and Culex mosquito species, which naturally harbor CI-inducing Wolbachia, transinfection can be achieved by either introducing a novel strain to generate superinfection or replacing the native infection with a different Wolbachia strain in a symbiont-free line, which is derived from antibiotic treatment of the wild type. Here, we use A. aegypti as an example to describe the Wolbachia transinfection method, which can be adapted to other insect species, such as planthoppers, according to their specific developmental requirements.}, } @article {pmid38006551, year = {2024}, author = {Fallon, AM}, title = {Preparation of Infectious Wolbachia from a Mosquito Cell Line.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {157-171}, pmid = {38006551}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; Mitomycin ; Cell Line ; *Aedes ; }, abstract = {Eventual genetic engineering of Wolbachia will require maximizing recovery of infectious bacteria, maintaining Wolbachia in a viable state for efficient manipulation, and reinfection of host cells for propagation and expansion of recombinant progeny. Challenges to manipulating Wolbachia arise from its obligate intracellular lifestyle and inability to divide outside a host cell, requiring modifications of standard bacteriological methods. The Aedes albopictus C7-10 cell line has proven to be a good recipient for the Wolbachia supergroup B strain, wStri, from the planthopper Laodelphax striatellus; the persistently infected C/wStri1 population provides a source of wStri inoculum that can be used systematically to explore conditions that increase yields of infectious material from input Wolbachia and identify conditions conducive to Wolbachia replication. After reintroduction into naive, uninfected C7-10 cells, wStri recovery, relative to the input inoculum, is influenced by diverse conditions, such as the cell cycle arrest that follows treatment of infected host cells with the insect steroid hormone, 20-hydroxyecdysone. Pretreatment of recipient cells with mitomycin C, which cross-links DNA and inhibits host cell replication, can improve recovery from low levels of input Wolbachia. This protocol describes preparation of infectious inoculum from Aedes albopictus C/wStri1 cells and amplification of Wolbachia in mitomycin C-treated, uninfected C7-10 cells, followed by a brief description of conditions used for various small-scale manipulations of Wolbachia in infected cells.}, } @article {pmid38006550, year = {2024}, author = {Bishop, C and Asgari, S}, title = {Use of Antibiotics to Eliminate Wolbachia from Mosquitoes and Cell Culture.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {147-156}, pmid = {38006550}, issn = {1940-6029}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Wolbachia/genetics ; *Aedes ; Cell Line ; Cell Culture Techniques ; }, abstract = {Removal of Wolbachia from infected insects is required in most experimental settings when the effects of Wolbachia on biological traits, pathogen blocking, reproduction, and fitness are assessed. This is to ensure that the genetic backgrounds of Wolbachia-infected and uninfected insects are the same. Here, we describe methodologies used for the elimination of Wolbachia from insects and insect cell lines with antibiotics.}, } @article {pmid38006549, year = {2024}, author = {Madhav, M and James, P}, title = {Developing Wolbachia-Infected Cell Lines from Insects.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {137-145}, pmid = {38006549}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia ; Insecta ; Cell Line ; }, abstract = {Wolbachia, an intracellular bacterium of arthropods, is an ideal candidate for use in the biological control of insect pests. The inability of Wolbachia to grow in the extracellular environment requires maintenance in live insects or insect cell cultures. Growing and adapting high-density Wolbachia in the targeted host cells improves the possibility of cross-species transinfection. Here, we describe a method for the establishment of a primary cell culture from insect embryos and its transinfection with Wolbachia.}, } @article {pmid38006548, year = {2024}, author = {Kaur, A and Brown, AMV}, title = {Detection and Analysis of Wolbachia in Plant-Parasitic Nematodes and Insights into Wolbachia Evolution.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {115-134}, pmid = {38006548}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Nematoda/genetics ; Plants/parasitology ; Genomics ; Metagenomics ; }, abstract = {Since the discovery of Wolbachia in plant-parasitic nematodes (PPNs), there has been increased interest in this earliest branching clade that may hold important clues to early transitions in Wolbachia function in the Ecdysozoa. However, due to the specialized skills and equipment of nematology and the difficulty in culturing most PPNs, these PPN-type Wolbachia remain undersampled and poorly understood. To date, there are few established laboratory methods for working with PPN-type Wolbachia strains, and most research has relied on chance discovery and comparative genomics. Here, we address this challenge by providing detailed methods to assist researchers with more efficiently collecting PPNs and screen these communities, populations, or single nematodes with a newly developed PPN-type Wolbachia-specific PCR assay. We provide an overview of the typical yields and outcomes of these methods, to facilitate further targeted cultivation or experimental methods, and finally we provide a short introduction to some of the specific challenges and solutions in following through with comparative or population genomics on PPN-type Wolbachia strains.}, } @article {pmid38006547, year = {2024}, author = {Kakumanu, ML and Hickin, ML and Schal, C}, title = {Detection, Quantification, and Elimination of Wolbachia in Bed Bugs.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {97-114}, pmid = {38006547}, issn = {1940-6029}, mesh = {Animals ; *Bedbugs/genetics ; *Wolbachia/genetics ; Anti-Bacterial Agents ; }, abstract = {Wolbachia is an obligatory nutritional symbiont of the common bed bug, Cimex lectularius, providing B-vitamins to its host. The biological significance of Wolbachia to bed bugs is investigated primarily by eliminating the symbiont with antibiotics, which is followed by confirmation with molecular assays. In this chapter, we describe a protocol for eliminating Wolbachia in bed bugs using the ansamycin antibiotic rifampicin (also known as rifampin) and three molecular methods to accurately detect and quantify the Wolbachia gene copies in bed bug samples. We describe the digital droplet PCR (ddPCR), a highly sensitive technique for absolute quantification of low abundance target genes, which has proven to be a valuable technique for confirmation of the elimination of Wolbachia.}, } @article {pmid38006546, year = {2024}, author = {Bertaux, J and Raimond, M and Grève, P}, title = {Pillbug: A Toolbox for Feminisation.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {69-95}, pmid = {38006546}, issn = {1940-6029}, mesh = {Animals ; Male ; Humans ; Female ; *Isopoda/genetics/metabolism ; Feminization ; *Butterflies ; Sex Differentiation ; Androgens/metabolism ; *Wolbachia ; }, abstract = {Wolbachia-induced feminisation is an extended phenotype that has been observed in only three models: isopods, butterflies and leafhoppers. Even among these, the process of feminisation differs markedly, especially since sexual differentiation is coordinated body-wide by a hormone in isopods, whereas in insects it is a chromosomally determined process and enforced cell by cell. For isopods, Armadillidium vulgare has contributed substantially to an understanding of Wolbachia-mediated feminisation, because a wide array of know-how and methods has been tailored to this model. Here, we describe establishment and maintenance of Wolbachia-infected laboratory lineages from animals collected in the field, the grafting of an androgenic gland to demonstrate the mode of action of Wolbachia (i.e. the disruption of the Androgenic Hormone pathway), and transinfection of naïve adults with Wolbachia. Finally, we describe selection of a ZZ genetic background in lineages created from transinfected WZ females, which is necessary to benefit from a property intrinsic to the naturally infected lineages: In a ZZ background, the vertical transmission rate of Wolbachia directly equates to the female ratio in the progeny. These protocols provide the essential framework for investigating molecular aspects of Wolbachia-induced feminisation.}, } @article {pmid38006545, year = {2024}, author = {Fricke, LC and Lindsey, ARI}, title = {Examining Wolbachia-Induced Parthenogenesis in Hymenoptera.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {55-68}, pmid = {38006545}, issn = {1940-6029}, mesh = {Animals ; Male ; Female ; *Wolbachia/genetics ; Parthenogenesis/genetics ; *Wasps/genetics ; Reproduction ; }, abstract = {The maternally transmitted reproductive manipulator Wolbachia can impact sex ratios of its arthropod host by different mechanisms, ultimately promoting the spread of infection across a population. One of these reproductive phenotypes, parthenogenesis induction (PI), is characterized by the asexual production of female offspring, which in many cases results in an entirely female population. Cases of Wolbachia-mediated PI are most common in the order Hymenoptera, specifically in parasitoid wasps. The complex sex determination pathways of hymenopterans, their diverse life histories, the multiple cytogenetic mechanisms of PI, and the lack of males make functional studies of parthenogenesis induction challenging. Here, we describe the mechanisms of PI, outline methods to recognize and cure PI-Wolbachia infection, and note possible complications when working with PI-Wolbachia strains and their parthenogenetic hosts.}, } @article {pmid38006544, year = {2024}, author = {Schedl, ME and Nguyen, NH and Unckless, RL and Perlmutter, JI}, title = {Maintenance and Evaluation of Wolbachia Male-Killers of Dipterans.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {29-53}, pmid = {38006544}, issn = {1940-6029}, mesh = {Male ; Animals ; *Wolbachia ; Drosophila ; Sex Ratio ; Embryonic Development ; }, abstract = {As research on Wolbachia male-killers of dipterans increases over time, several standard methods have emerged for the validation and characterization of these Wolbachia strains. Here, we describe typical steps for maintenance and evaluation of male-killers in the laboratory. Drosophila that host Wolbachia male-killers include both mushroom-feeders and flies reared on standard media, which require different maintenance protocols. In addition, male killing in flies typically occurs during embryonic development, and as such, many experiments require embryo tissue samples. We provide step-by-step instructions for standard methods of isolation, rearing, confirmation of Wolbachia male-killing as the basis of a sex ratio bias, and collection of embryos from these species. Thus, anyone intending to study a male-killer in the lab will have the complete set of protocols necessary to collect and rear them, validate them, and collect embryonic tissue for further study.}, } @article {pmid38006543, year = {2024}, author = {Namias, A and Atyame, C and Pasteur, N and Sicard, M and Weill, M}, title = {Investigation of Cytoplasmic Incompatibility Patterns in Culex pipiens: From Field Samples to Laboratory Isofemale Lines.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {17-27}, pmid = {38006543}, issn = {1940-6029}, mesh = {Animals ; *Culex/genetics ; Cytoplasm ; Cytosol ; }, abstract = {This protocol describes approaches to qualify Wolbachia-induced cytoplasmic incompatibility (CI) patterns (compatible, uni or bidirectional) in crosses between two or more Culex pipiens isofemale lines, hosting different Wolbachia (wPip) strains. Here, we describe how to (1) collect the larvae in the field and grow them to the adult stage in the insectary, (2) set up isofemale lines in the insectary, (3) genetically characterize the wPip group of these lines, and (4) perform reciprocal crosses to qualify CI.}, } @article {pmid38006542, year = {2024}, author = {Fallon, AM}, title = {Wolbachia: Advancing into a Second Century.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {1-13}, pmid = {38006542}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Filarioidea ; *Arthropods ; Symbiosis ; }, abstract = {Wolbachia pipientis had its scientific debut nearly a century ago and has recently emerged as a target for therapeutic treatment of filarial infections and an attractive tool for control of arthropod pests. Wolbachia was known as a biological entity before DNA was recognized as the molecule that carries the genetic information on which life depends, and before arthropods and nematodes were grouped in the Ecdysozoa. Today, some investigators consider Wolbachia the most abundant endosymbiont on earth, given the numbers of its hosts and its diverse mutualistic, commensal, and parasitic roles in their life histories. Recent advances in molecular technologies have revolutionized our understanding of Wolbachia and its associated reproductive phenotypes. New models have emerged for its investigation, and substantial progress has been made towards Wolbachia-based interventions in medicine and agriculture. Here I introduce Wolbachia, with a focus on aspects of its biology that are covered in greater detail in subsequent chapters.}, } @article {pmid38003752, year = {2023}, author = {Garrigós, M and Garrido, M and Panisse, G and Veiga, J and Martínez-de la Puente, J}, title = {Interactions between West Nile Virus and the Microbiota of Culex pipiens Vectors: A Literature Review.}, journal = {Pathogens (Basel, Switzerland)}, volume = {12}, number = {11}, pages = {}, pmid = {38003752}, issn = {2076-0817}, support = {PID2020-118205GB-I00//Ministerio Español de Ciencia e Innovación/ ; PRE2021-098544//Ministerio Español de Ciencia e Innovación/ ; FJC2021-048057-I//Ministerio Español de Ciencia e Innovación/ ; María Zambrano//Ministerio Español de Universidades/ ; Margarita Salas//Ministerio Español de Universidades/ ; P21_00049//Junta de Andalucía, Consejería de Universidad, Investigación e Innovación/ ; }, abstract = {The flavivirus West Nile virus (WNV) naturally circulates between mosquitoes and birds, potentially affecting humans and horses. Different species of mosquitoes play a role as vectors of WNV, with those of the Culex pipiens complex being particularly crucial for its circulation. Different biotic and abiotic factors determine the capacity of mosquitoes for pathogen transmission, with the mosquito gut microbiota being recognized as an important one. Here, we review the published studies on the interactions between the microbiota of the Culex pipiens complex and WNV infections in mosquitoes. Most articles published so far studied the interactions between bacteria of the genus Wolbachia and WNV infections, obtaining variable results regarding the directionality of this relationship. In contrast, only a few studies investigate the role of the whole microbiome or other bacterial taxa in WNV infections. These studies suggest that bacteria of the genera Serratia and Enterobacter may enhance WNV development. Thus, due to the relevance of WNV in human and animal health and the important role of mosquitoes of the Cx. pipiens complex in its transmission, more research is needed to unravel the role of mosquito microbiota and those factors affecting this microbiota on pathogen epidemiology. In this respect, we finally propose future lines of research lines on this topic.}, } @article {pmid37999039, year = {2023}, author = {El Yamlahi, Y and Bel Mokhtar, N and Maurady, A and Britel, MR and Batargias, C and Mutembei, DE and Nyingilili, HS and Malulu, DJ and Malele, II and Asimakis, E and Stathopoulou, P and Tsiamis, G}, title = {Characterization of the Bacterial Profile from Natural and Laboratory Glossina Populations.}, journal = {Insects}, volume = {14}, number = {11}, pages = {}, pmid = {37999039}, issn = {2075-4450}, support = {URT-22658//FAO/IAEA Agriculture and Biotechnology Laboratories/ ; }, abstract = {Tsetse flies (Glossina spp.; Diptera: Glossinidae) are viviparous flies that feed on blood and are found exclusively in sub-Saharan Africa. They are the only cyclic vectors of African trypanosomes, responsible for human African trypanosomiasis (HAT) and animal African trypanosomiasis (AAT). In this study, we employed high throughput sequencing of the 16S rRNA gene to unravel the diversity of symbiotic bacteria in five wild and three laboratory populations of tsetse species (Glossina pallidipes, G. morsitans, G. swynnertoni, and G. austeni). The aim was to assess the dynamics of bacterial diversity both within each laboratory and wild population in relation to the developmental stage, insect age, gender, and location. Our results indicated that the bacterial communities associated with the four studied Glossina species were significantly influenced by their region of origin, with wild samples being more diverse compared to the laboratory samples. We also observed that the larval microbiota was significantly different than the adults. Furthermore, the sex and the species did not significantly influence the formation of the bacterial profile of the laboratory colonies once these populations were kept under the same rearing conditions. In addition, Wigglesworthia, Acinetobacter, and Sodalis were the most abundant bacterial genera in all the samples, while Wolbachia was significantly abundant in G. morsitans compared to the other studied species. The operational taxonomic unit (OTU) co-occurrence network for each location (VVBD insectary, Doma, Makao, and Msubugwe) indicated a high variability between G. pallidipes and the other species in terms of the number of mutual exclusion and copresence interactions. In particular, some bacterial genera, like Wigglesworthia and Sodalis, with high relative abundance, were also characterized by a high degree of interactions.}, } @article {pmid37995370, year = {2023}, author = {Strunov, A and Kirchner, S and Schindelar, J and Kruckenhauser, L and Haring, E and Kapun, M}, title = {Historic Museum Samples Provide Evidence for a Recent Replacement of Wolbachia Types in European Drosophila melanogaster.}, journal = {Molecular biology and evolution}, volume = {40}, number = {12}, pages = {}, pmid = {37995370}, issn = {1537-1719}, support = {P 32275/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Museums ; Biological Evolution ; Reproduction ; Symbiosis ; }, abstract = {Wolbachia is one of the most common bacterial endosymbionts, which is frequently found in numerous arthropods and nematode taxa. Wolbachia infections can have a strong influence on the evolutionary dynamics of their hosts since these bacteria are reproductive manipulators that affect the fitness and life history of their host species for their own benefit. Host-symbiont interactions with Wolbachia are perhaps best studied in the model organism Drosophila melanogaster, which is naturally infected with at least 5 different variants among which wMel and wMelCS are the most frequent ones. Comparisons of infection types between natural flies and long-term lab stocks have previously indicated that wMelCS represents the ancestral type, which was only very recently replaced by the nowadays dominant wMel in most natural populations. In this study, we took advantage of recently sequenced museum specimens of D. melanogaster that have been collected 90 to 200 yr ago in Northern Europe to test this hypothesis. Our comparison to contemporary Wolbachia samples provides compelling support for the replacement hypothesis. Our analyses show that sequencing data from historic museum specimens and their bycatch are an emerging and unprecedented resource to address fundamental questions about evolutionary dynamics in host-symbiont interactions. However, we also identified contamination with DNA from crickets that resulted in co-contamination with cricket-specific Wolbachia in several samples. These results underpin the need for rigorous quality assessments of museomic data sets to account for contamination as a source of error that may strongly influence biological interpretations if it remains undetected.}, } @article {pmid37994906, year = {2023}, author = {Meyer, DF and Moumène, A and Rodrigues, V}, title = {Microbe Profile: Ehrlichia ruminantium - stealthy as it goes.}, journal = {Microbiology (Reading, England)}, volume = {169}, number = {11}, pages = {}, pmid = {37994906}, issn = {1465-2080}, mesh = {*Ehrlichia ruminantium ; }, abstract = {Ehrlichia ruminantium is an obligate intracellular pathogenic bacterium that causes heartwater, a fatal disease of ruminants in tropical areas. Some human cases have also been reported. This globally important pathogen is primarily transmitted by ticks of the Amblyomma genus and threatens American mainland. E. ruminantium replicates within eukaryotic mammal or tick cell is a membrane-bound vacuole, where it undergoes a biphasic developmental growth cycle and differentiates from noninfectious replicative form into infectious elementary bodies. The ability of E. ruminantium to hijack host cellular processes and avoid innate immunity is a fundamental, but not yet fully understood, virulence trait of this stealth pathogen in the genomic era.}, } @article {pmid37989350, year = {2023}, author = {Bannister-Tyrrell, M and Hillman, A and Indriani, C and Ahmad, RA and Utarini, A and Simmons, CP and Anders, KL and Sergeant, E}, title = {Utility of surveillance data for planning for dengue elimination in Yogyakarta, Indonesia: a scenario-tree modelling approach.}, journal = {BMJ global health}, volume = {8}, number = {11}, pages = {}, pmid = {37989350}, issn = {2059-7908}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Humans ; *Dengue Virus ; Indonesia/epidemiology ; *Aedes/microbiology ; Incidence ; *Dengue/epidemiology/prevention & control ; }, abstract = {INTRODUCTION: Field trials and modelling studies suggest that elimination of dengue transmission may be possible through widespread release of Aedes aegypti mosquitoes infected with the insect bacterium Wolbachia pipientis (wMel strain), in conjunction with routine dengue control activities. This study aimed to develop a modelling framework to guide planning for the potential elimination of locally acquired dengue in Yogyakarta, a city of almost 400 000 people in Java, Indonesia.

METHODS: A scenario-tree modelling approach was used to estimate the sensitivity of the dengue surveillance system (including routine hospital-based reporting and primary-care-based enhanced surveillance), and time required to demonstrate elimination of locally acquired dengue in Yogyakarta city, assuming the detected incidence of dengue decreases to zero in the future. Age and gender were included as risk factors for dengue, and detection nodes included the probability of seeking care, probability of sample collection and testing, diagnostic test sensitivity and probability of case notification. Parameter distributions were derived from health system data or estimated by expert opinion. Alternative simulations were defined based on changes to key parameter values, separately and in combination.

RESULTS: For the default simulation, median surveillance system sensitivity was 0.131 (95% PI 0.111 to 0.152) per month. Median confidence in dengue elimination reached 80% after a minimum of 13 months of zero detected dengue cases and 90% confidence after 25 months, across different scenarios. The alternative simulations investigated produced relatively small changes in median system sensitivity and time to elimination.

CONCLUSION: This study suggests that with a combination of hospital-based surveillance and enhanced clinic-based surveillance for dengue, an acceptable level of confidence (80% probability) in the elimination of locally acquired dengue can be reached within 2 years. Increasing the surveillance system sensitivity could shorten the time to first ascertainment of elimination of dengue and increase the level of confidence in elimination.}, } @article {pmid37985236, year = {2023}, author = {Fukui, T and Kiuchi, T and Tomihara, K and Muro, T and Matsuda-Imai, N and Katsuma, S}, title = {Expression of the Wolbachia male-killing factor Oscar impairs dosage compensation in lepidopteran embryos.}, journal = {FEBS letters}, volume = {}, number = {}, pages = {}, doi = {10.1002/1873-3468.14777}, pmid = {37985236}, issn = {1873-3468}, support = {2023//G-7 Scholarship Foundation/ ; 17H06431//Japan Society for the Promotion of Science/ ; 22H00366//Japan Society for the Promotion of Science/ ; JP21J12325//Japan Society for the Promotion of Science/ ; }, abstract = {Wolbachia are intracellular bacteria in insects that can manipulate the sexual development and reproduction by male killing or other methods. We have recently identified a Wolbachia protein named Oscar that acts as a male-killing factor for lepidopteran insects. Oscar interacts with the Masculinizer (Masc) protein, which is required for both masculinization and dosage compensation (DC) in lepidopteran insects. Embryonic expression of Oscar inhibits masculinization and causes male killing in two lepidopteran species, Ostrinia furnacalis and Bombyx mori. However, it remains unknown whether Oscar-induced male killing is caused by a failure of DC. Here, we performed a transcriptome analysis of Oscar complementary RNA-injected O. furnacalis and B. mori embryos, and found that Oscar primarily targets the Masc protein, resulting in male killing by interfering with DC in lepidopteran insects.}, } @article {pmid37978413, year = {2023}, author = {Pascar, J and Middleton, H and Dorus, S}, title = {Aedes aegypti microbiome composition covaries with the density of Wolbachia infection.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {255}, pmid = {37978413}, issn = {2049-2618}, support = {R21 HD088910/HD/NICHD NIH HHS/United States ; }, mesh = {Humans ; Animals ; Female ; *Dengue Virus ; *Aedes ; *Wolbachia/genetics ; Mosquito Vectors/microbiology ; Drosophila melanogaster/microbiology ; *Microbiota ; }, abstract = {BACKGROUND: Wolbachia is a widespread bacterial endosymbiont that can inhibit vector competency when stably transinfected into the mosquito, Aedes aegypti, a primary vector of the dengue virus (DENV) and other arboviruses. Although a complete mechanistic understanding of pathogen blocking is lacking, it is likely to involve host immunity induction and resource competition between Wolbachia and DENV, both of which may be impacted by microbiome composition. The potential impact of Wolbachia transinfection on host fitness is also of importance given the widespread release of mosquitos infected with the Drosophila melanogaster strain of Wolbachia (wMel) in wild populations. Here, population-level genomic data from Ae. aegypti was surveyed to establish the relationship between the density of wMel infection and the composition of the host microbiome.

RESULTS: Analysis of genomic data from 172 Ae. aegypti females across six populations resulted in an expanded and quantitatively refined, species-level characterization of the bacterial, archaeal, and fungal microbiome. This included 844 species of bacteria across 23 phyla, of which 54 species were found to be ubiquitous microbiome members across these populations. The density of wMel infection was highly variable between individuals and negatively correlated with microbiome diversity. Network analyses revealed wMel as a hub comprised solely of negative interactions with other bacterial species. This contrasted with the large and highly interconnected network of other microbiome species that may represent members of the midgut microbiome community in this population.

CONCLUSION: Our bioinformatic survey provided a species-level characterization of Ae. aegypti microbiome composition and variation. wMel load varied substantially across populations and individuals and, importantly, wMel was a major hub of a negative interactions across the microbiome. These interactions may be an inherent consequence of heightened pathogen blocking in densely infected individuals or, alternatively, may result from antagonistic Wolbachia-incompatible bacteria that could impede the efficacy of wMel as a biological control agent in future applications. The relationship between wMel infection variation and the microbiome warrants further investigation in the context of developing wMel as a multivalent control agent against other arboviruses. Video Abstract.}, } @article {pmid37968789, year = {2023}, author = {Valdivia, C and Newton, JA and von Beeren, C and O'Donnell, S and Kronauer, DJC and Russell, JA and Łukasik, P}, title = {Microbial symbionts are shared between ants and their associated beetles.}, journal = {Environmental microbiology}, volume = {25}, number = {12}, pages = {3466-3483}, doi = {10.1111/1462-2920.16544}, pmid = {37968789}, issn = {1462-2920}, support = {//"Excellence Initiative - Research University" at the Faculty of Biology of the Jagiellonian University in Kraków, Poland/ ; PPN/PPO/2018/1/00015//Narodowa Agencja Wymiany Akademickiej/ ; 2018/31/B/NZ8/01158//Narodowe Centrum Nauki/ ; 1050360//National Science Foundation/ ; }, mesh = {Animals ; *Ants/genetics/microbiology ; Phylogeny ; *Coleoptera ; RNA, Ribosomal, 16S/genetics ; Larva ; *Microbiota ; Bacteria/genetics ; Symbiosis ; }, abstract = {The transmission of microbial symbionts across animal species could strongly affect their biology and evolution, but our understanding of transmission patterns and dynamics is limited. Army ants (Formicidae: Dorylinae) and their hundreds of closely associated insect guest species (myrmecophiles) can provide unique insights into interspecific microbial symbiont sharing. Here, we compared the microbiota of workers and larvae of the army ant Eciton burchellii with those of 13 myrmecophile beetle species using 16S rRNA amplicon sequencing. We found that the previously characterized specialized bacterial symbionts of army ant workers were largely absent from ant larvae and myrmecophiles, whose microbial communities were usually dominated by Rickettsia, Wolbachia, Rickettsiella and/or Weissella. Strikingly, different species of myrmecophiles and ant larvae often shared identical 16S rRNA genotypes of these common bacteria. Protein-coding gene sequences confirmed the close relationship of Weissella strains colonizing army ant larvae, some workers and several myrmecophile species. Unexpectedly, these strains were also similar to strains infecting dissimilar animals inhabiting very different habitats: trout and whales. Together, our data show that closely interacting species can share much of their microbiota, and some versatile microbial species can inhabit and possibly transmit across a diverse range of hosts and environments.}, } @article {pmid37957741, year = {2023}, author = {Moldovan, OT and Carrell, AA and Bulzu, PA and Levei, E and Bucur, R and Sitar, C and Faur, L and Mirea, IC and Șenilă, M and Cadar, O and Podar, M}, title = {The gut microbiome mediates adaptation to scarce food in Coleoptera.}, journal = {Environmental microbiome}, volume = {18}, number = {1}, pages = {80}, pmid = {37957741}, issn = {2524-6372}, support = {R01 DE024463/DE/NIDCR NIH HHS/United States ; R01DE024463/NH/NIH HHS/United States ; }, abstract = {Beetles are ubiquitous cave invertebrates worldwide that adapted to scarce subterranean resources when they colonized caves. Here, we investigated the potential role of gut microbiota in the adaptation of beetles to caves from different climatic regions of the Carpathians. The beetles' microbiota was host-specific, reflecting phylogenetic and nutritional adaptation. The microbial community structure further resolved conspecific beetles by caves suggesting microbiota-host coevolution and influences by local environmental factors. The detritivore species hosted a variety of bacteria known to decompose and ferment organic matter, suggesting turnover and host cooperative digestion of the sedimentary microbiota and allochthonous-derived nutrients. The cave Carabidae, with strong mandibula, adapted to predation and scavenging of animal and plant remains, had distinct microbiota dominated by symbiotic lineages Spiroplasma or Wolbachia. All beetles had relatively high levels of fermentative Carnobacterium and Vagococcus involved in lipid accumulation and a reduction of metabolic activity, and both features characterize adaptation to caves.}, } @article {pmid37952351, year = {2024}, author = {El Hamss, H and Maruthi, MN and Omongo, CA and Wang, HL and van Brunschot, S and Colvin, J and Delatte, H}, title = {Microbiome diversity and composition in Bemisia tabaci SSA1-SG1 whitefly are influenced by their host's life stage.}, journal = {Microbiological research}, volume = {278}, number = {}, pages = {127538}, doi = {10.1016/j.micres.2023.127538}, pmid = {37952351}, issn = {1618-0623}, mesh = {Animals ; *Hemiptera/microbiology ; *Microbiota ; Life Cycle Stages ; }, abstract = {Within the Bemisia tabaci group of cryptic whitefly species, many are damaging agricultural pests and plant-virus vectors, conferring upon this group the status of one of the world's top 100 most invasive and destructive species, affecting farmers' income and threatening their livelihoods. Studies on the microbiome of whitefly life stages are scarce, although their composition and diversity greatly influence whitefly fitness and development. We used high-throughput sequencing to understand microbiome diversity in different developmental stages of the B. tabaci sub-Saharan Africa 1 (SSA1-SG1) species of the whitefly from Uganda. Endosymbionts (Portiera, Arsenophonus, Wolbachia, and Hemipteriphilus were detected but excluded from further statistical analysis as they were not influenced by life stage using Permutational Multivariate Analysis of Variance Using Distance Matrices (ADONIS, p = 0.925 and Bray, p = 0.903). Our results showed significant differences in the meta microbiome composition in different life stages of SSA1-SG1. The diversity was significantly higher in eggs (Shannon, p = 0.024; Simpson, p = 0.047) than that in nymphs and pupae, while the number of microbial species observed by the amplicon sequence variant (ASV) was not significant (n(ASV), p = 0.094). At the phylum and genus levels, the dominant constituents in the microbiome changed significantly during various developmental stages, with Halomonas being present in eggs, whereas Bacillus and Caldalkalibacillus were consistently found across all life stages. These findings provide the first description of differing meta microbiome diversity in the life stage of whiteflies, suggesting their putative role in whitefly development.}, } @article {pmid37949964, year = {2024}, author = {Bickerstaff, JRM and Jordal, BH and Riegler, M}, title = {Two sympatric lineages of Australian Cnestus solidus share Ambrosiella symbionts but not Wolbachia.}, journal = {Heredity}, volume = {132}, number = {1}, pages = {43-53}, pmid = {37949964}, issn = {1365-2540}, abstract = {Sympatric lineages of inbreeding species provide an excellent opportunity to investigate species divergence patterns and processes. Many ambrosia beetle lineages (Curculionidae: Scolytinae) reproduce by predominant inbreeding through sib mating in nests excavated in woody plant parts wherein they cultivate symbiotic ambrosia fungi as their sole source of nutrition. The Xyleborini ambrosia beetle species Cnestus solidus and Cnestus pseudosolidus are sympatrically distributed across eastern Australia and have overlapping morphological variation. Using multilocus sequencing analysis of individuals collected from 19 sites spanning their sympatric distribution, we assessed their phylogenetic relationships, taxonomic status and microbial symbionts. We found no genetic differentiation between individuals morphologically identified as C. solidus and C. pseudosolidus confirming previous suggestions that C. pseudosolidus is synonymous to C. solidus. However, within C. solidus we unexpectedly discovered the sympatric coexistence of two morphologically indistinguishable but genetically distinct lineages with small nuclear yet large mitochondrial divergence. At all sites except one, individuals of both lineages carried the same primary fungal symbiont, a new Ambrosiella species, indicating that fungal symbiont differentiation may not be involved in lineage divergence. One strain of the maternally inherited bacterial endosymbiont Wolbachia was found at high prevalence in individuals of the more common lineage but not in the other, suggesting that it may influence host fitness. Our data suggest that the two Australian Cnestus lineages diverged allopatrically, and one lineage then acquired Wolbachia. Predominant inbreeding and Wolbachia infection may have reinforced reproductive barriers between these two lineages after their secondary contact contributing to their current sympatric distribution.}, } @article {pmid37948354, year = {2023}, author = {Hussain, M and Zhong, Y and Tao, T and Xiu, B and Ye, F and Gao, J and Mao, R}, title = {Effect of tree height and spraying methods on Diaphorina citri kuwayama endosymbionts in the context of Huanglongbing disease management in citrus orchards.}, journal = {Pest management science}, volume = {}, number = {}, pages = {}, doi = {10.1002/ps.7880}, pmid = {37948354}, issn = {1526-4998}, support = {2021GDASYL-20210103051//The GDAS Special Project of Science and Technology Development/ ; 2022GDASZH-2022030501//The GDAS Special Project of Science and Technology Development/ ; QN2022030005//Foreign Youth Talent Programs/ ; 2023SDZG06//the open competition program of top ten critical priorities of Agricultural Science and Technology Innovation for the 14th Five-year plan of Guangdong province/ ; 2022SDZG06//the open competition program of top ten critical priorities of Agricultural Science and Technology Innovation for the 14th Five-year plan of Guangdong province/ ; 20220610043//the Research Programs of Guangzhou/ ; 202103000065//the Research Programs of Guangzhou/ ; KTP20210352//Science and Technology Planning Project of Guangdong Province/ ; }, abstract = {BACKGROUND: Huanglongbing (HLB) (caused by Candidatus Liberibacter asiaticus) is the most damaging disease of citrus around the world. This study investigated the effects of citrus tree height on Diaphorina citri Kuwayama mortality, endosymbiont responses, and HLB distribution.

RESULTS: The results reveal that the age of citrus trees plays a significant role in psyllid mortality. Interestingly, the cumulative mean mortality (%) of psyllids over the seven-day observation period was higher (31.50±0.03) when four-year-old (501A1, 502A2, 501A3) citrus trees were sprayed with a US-SMART mechanical sprayer. In contrast, the psyllids mortality was 0.09±0.23 for the 13-year-old citrus trees (104A2, 104A3, 104C1) sprayed with a US-SMART mechanical sprayer and 9.10±0.05 for 13-year-old (502A2, 502B2, 502D1) citrus trees sprayed with a fixed US-SMART mechanical sprayer. Our findings also revealed that psyllids from both four- and 13-year-old citrus trees carried Candidatus Carsonella ruddii species and Wolbachia, the primary and secondary endosymbionts, respectively. Surprisingly, infection rates of these endosymbionts remained consistent across different age groups, as confirmed by quantitative polymerase chain reaction analysis. Furthermore, our study highlights the significance of tree height as a proxy for tree age in influencing HLB occurrence. Specifically, four-year-old citrus trees subjected to the US-SMART mechanical sprayer for citrus psyllid control demonstrated effective disease management compared to 13-year-old (104A2, 104A3, 104C1) citrus trees sprayed with US-SMART mechanical sprayers. Additionally, the investigation explored the impact of tree height on HLB distribution. In four-year-old trees, no significant correlation between HLB disease and tree height was observed, potentially due to effective spray coverage with US-SMART mechanical sprayer. However, in 13-year-old (104A2, 104A3, 104C1) citrus tree sprayed with US-SMART mechanical sprayer, a positive correlation between tree height and HLB disease was evident.

CONCLUSION: This research provides valuable insights into the complex interaction between citrus tree age, psyllid endosymbionts responses, and HLB distribution. These results emphasize effective HLB management strategies, especially in orchards with diverse tree age populations, ultimately contributing to the long-term sustainability of citrus cultivation. © 2023 Society of Chemical Industry.}, } @article {pmid37941933, year = {2023}, author = {Caamal-Chan, MG and Barraza, A and Loera-Muro, A and Montes-Sánchez, JJ and Castellanos, T and Rodríguez-Pagaza, Y}, title = {Bacterial communities of the psyllid pest Bactericera cockerelli (Hemiptera: Triozidae) Central haplotype of tomato crops cultivated at different locations of Mexico.}, journal = {PeerJ}, volume = {11}, number = {}, pages = {e16347}, pmid = {37941933}, issn = {2167-8359}, mesh = {Animals ; Haplotypes ; *Solanum lycopersicum ; RNA, Ribosomal, 16S/genetics ; *Hemiptera/genetics ; Phylogeny ; Mexico ; Bacteria/genetics ; Liberibacter/genetics ; Crops, Agricultural/genetics ; }, abstract = {BACKGROUND: The psyllid, Bactericera cockerelli, is an insect vector of 'Candidatus Liberibacter' causing "Zebra chip" disease that affects potato and other Solanaceae crops worldwide. In the present study, we analyzed the bacterial communities associated with the insect vector Bactericera cockerelli central haplotype of tomato crop fields in four regions from Mexico.

METHODS: PCR was used to amplify the mitochondrial cytochrome oxidase I gene (mtCOI) and then analyze the single nucleotide polymorphisms (SNP) and phylogenetic analysis for haplotype identification of the isolated B. cockerelli. Moreover, we carried out the microbial diversity analysis of several B. cockerelli collected from four regions of Mexico through the NGS sequencing of 16S rRNA V3 region. Finally, Wolbachia was detected by the wsp gene PCR amplification, which is the B. cockerelli facultative symbiont. Also we were able to confirm the relationship with several Wolbachia strains by phylogenetic analysis.

RESULTS: Our results pointed that B. cockerelli collected in the four locations from Mexico (Central Mexico: Queretaro, and Northern Mexico: Sinaloa, Coahuila, and Nuevo Leon) were identified, such as the central haplotype. Analyses of the parameters of the composition, relative abundance, and diversity (Shannon index: 1.328 ± 0.472; Simpson index 0.582 ± 0.167), showing a notably relatively few microbial species in B. cockerelli. Analyses identified various facultative symbionts, particularly the Wolbachia (Rickettsiales: Anaplasmataceae) with a relative abundance higher. In contrast, the genera of Sodalis and 'Candidatus Carsonella' (Gammaproteobacteria: Oceanospirillales: Halomonadaceae) were identified with a relatively low abundance. On the other hand, the relative abundance for the genus 'Candidatus Liberibacter' was higher only for some of the locations analyzed. PCR amplification of a fragment of the gene encoding a surface protein (wsp) of Wolbachia and phylogenetic analysis corroborated the presence of this bacterium in the central haplotype. Beta-diversity analysis revealed that the presence of the genus 'Candidatus Liberibacter' influences the microbiota structure of this psyllid species.

CONCLUSIONS: Our data support that the members with the highest representation in microbial community of B. cockerelli central haplotype, comprise their obligate symbiont, Carsonella, and facultative symbionts. We also found evidence that among the factors analyzed, the presence of the plant pathogen affects the structure and composition of the bacterial community associated with B. cockerelli.}, } @article {pmid37936139, year = {2023}, author = {Hakobyan, A and Velte, S and Sickel, W and Quandt, D and Stoll, A and Knief, C}, title = {Tillandsia landbeckii phyllosphere and laimosphere as refugia for bacterial life in a hyperarid desert environment.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {246}, pmid = {37936139}, issn = {2049-2618}, mesh = {Humans ; Soil Microbiology ; *Tillandsia ; Refugium ; *Microbiota ; Bacteria/genetics ; Plants/microbiology ; Soil ; Desert Climate ; }, abstract = {BACKGROUND: The lack of water is a major constraint for microbial life in hyperarid deserts. Consequently, the abundance and diversity of microorganisms in common habitats such as soil are strongly reduced, and colonization occurs primarily by specifically adapted microorganisms that thrive in particular refugia to escape the harsh conditions that prevail in these deserts. We suggest that plants provide another refugium for microbial life in hyperarid deserts. We studied the bacterial colonization of Tillandsia landbeckii (Bromeliaceae) plants, which occur in the hyperarid regions of the Atacama Desert in Chile, one of the driest and oldest deserts on Earth.

RESULTS: We detected clear differences between the bacterial communities being plant associated to those of the bare soil surface (PERMANOVA, R[2] = 0.187, p = 0.001), indicating that Tillandsia plants host a specific bacterial community, not only dust-deposited cells. Moreover, the bacterial communities in the phyllosphere were distinct from those in the laimosphere, i.e., on buried shoots (R[2] = 0.108, p = 0.001), indicating further habitat differentiation within plant individuals. The bacterial taxa detected in the phyllosphere are partly well-known phyllosphere colonizers, but in addition, some rather unusual taxa (subgroup2 Acidobacteriae, Acidiphilum) and insect endosymbionts (Wolbachia, "Candidatus Uzinura") were found. The laimosphere hosted phyllosphere-associated as well as soil-derived taxa. The phyllosphere bacterial communities showed biogeographic patterns across the desert (R[2] = 0.331, p = 0.001). These patterns were different and even more pronounced in the laimosphere (R[2] = 0.467, p = 0.001), indicating that different factors determine community assembly in the two plant compartments. Furthermore, the phyllosphere microbiota underwent temporal changes (R[2] = 0.064, p = 0.001).

CONCLUSIONS: Our data demonstrate that T. landbeckii plants host specific bacterial communities in the phyllosphere as well as in the laimosphere. Therewith, these plants provide compartment-specific refugia for microbial life in hyperarid desert environments. The bacterial communities show biogeographic patterns and temporal variation, as known from other plant microbiomes, demonstrating environmental responsiveness and suggesting that bacteria inhabit these plants as viable microorganisms. Video Abstract.}, } @article {pmid37928691, year = {2023}, author = {André, MR and Ikeda, P and Lee, DAB and do Amaral, RB and Carvalho, LAL and Pinheiro, DG and Torres, JM and de Mello, VVC and Rice, GK and Cer, RZ and Lourenço, EC and Oliveira, CE and Herrera, HM and Barros-Battesti, DM and Machado, RZ and Bishop-Lilly, KA and Dalgard, CL and Dumler, JS}, title = {Characterization of the bacterial microbiome of non-hematophagous bats and associated ectoparasites from Brazil.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1261156}, pmid = {37928691}, issn = {1664-302X}, abstract = {INTRODUCTION: Bats, along with their ectoparasites, harbor a wide diversity of symbiotic and potential pathogenic bacteria. Despite the enormous diversity of bats (181 species), few studies aimed to investigate the bacterial microbiome of Brazilian chiropterans and associated ectoparasites. This study aimed to characterize the bacterial microbiome of non-hematophagous bats and associated Streblidae flies and Macronyssidae and Spinturnicidae mites in the state of Mato Grosso do Sul, midwestern Brazil.

METHODS: Oral and rectal swabs were collected from 30 bats (Artibeus lituratus [n = 13], Artibeus planirostris [n  =  9], Eptesicus furinalis [n = 5], Carollia perspicillata [n = 2], and Platyrrhinus lineatus [n = 1]). In addition, a total of 58 mites (15 Macronyssidae and 43 Spinturnicidae) and 48 Streblidae bat flies were collected from the captured bats. After DNA extraction and purification, each sample's bacterial composition was analyzed with metagenomic sequencing.

RESULTS: The microbiome composition of both oral and rectal bat swab samples showed that Gammaproteobacteria was the most abundant bacterial class. Spiroplasma, Wolbachia and Bartonella represented the most abundant genera in Streblidae flies. While Wolbachia (Alphaproteobacteria) was the most abundant genus found in Spinturnicidae, Arsenophonus (Gammaproteobacteria) was found in high abundance in Macronyssidae mites. In addition to characterizing the microbiome of each sample at the class and genus taxonomic levels, we identified medically significant bacteria able to infect both animals and humans in oral (Streptococcus and Anaplasma) and rectal swabs (Enterobacter, Klebsiella, Escherichia, Enterococcus, Streptococcus), Macronyssidae (Anaplasma, Bartonella, Ehrlichia) and Spinturnicidae (Anaplasma, Bartonella) mites as well as Streblidae flies (Spiroplasma, Bartonella).

DISCUSSION AND CONCLUSION: Besides expanding the knowledge on the bacterial microbiome of non-hematophagous bats and Streblidae flies from Brazil, the present work showed, for the first time, the bacterial community of bat-associated Macronyssidae and Spinturnicidae mites.}, } @article {pmid37928198, year = {2023}, author = {Mee, L and Barribeau, SM}, title = {Influence of social lifestyles on host-microbe symbioses in the bees.}, journal = {Ecology and evolution}, volume = {13}, number = {11}, pages = {e10679}, pmid = {37928198}, issn = {2045-7758}, abstract = {Microbiomes are increasingly recognised as critical for the health of an organism. In eusocial insect societies, frequent social interactions allow for high-fidelity transmission of microbes across generations, leading to closer host-microbe coevolution. The microbial communities of bees with other social lifestyles are less studied, and few comparisons have been made between taxa that vary in social structure. To address this gap, we leveraged a cloud-computing resource and publicly available transcriptomic data to conduct a survey of microbial diversity in bee samples from a variety of social lifestyles and taxa. We consistently recover the core microbes of well-studied corbiculate bees, supporting this method's ability to accurately characterise microbial communities. We find that the bacterial communities of bees are influenced by host location, phylogeny and social lifestyle, although no clear effect was found for fungal or viral microbial communities. Bee genera with more complex societies tend to harbour more diverse microbes, with Wolbachia detected more commonly in solitary tribes. We present a description of the microbiota of Euglossine bees and find that they do not share the "corbiculate core" microbiome. Notably, we find that bacteria with known anti-pathogenic properties are present across social bee genera, suggesting that symbioses that enhance host immunity are important with higher sociality. Our approach provides an inexpensive means of exploring microbiomes of a given taxa and identifying avenues for further research. These findings contribute to our understanding of the relationships between bees and their associated microbial communities, highlighting the importance of considering microbiome dynamics in investigations of bee health.}, } @article {pmid37926475, year = {2023}, author = {Liu, Y and Wang, D and He, Z and Qian, D and Liu, Y and Yang, C and Lu, D and Zhang, H}, title = {[Molecular detection and phylogenetic analysis of Wolbachia infection in common mosquito species in Henan Province].}, journal = {Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control}, volume = {35}, number = {4}, pages = {389-393}, doi = {10.16250/j.32.1374.2023033}, pmid = {37926475}, issn = {1005-6661}, support = {LHGJ20220178//Henan Provincial Medical Science and Technology Research Project/ ; }, mesh = {Animals ; Humans ; Phylogeny ; *Wolbachia/genetics ; *Culex/genetics ; *Aedes/genetics ; DNA, Ribosomal ; }, abstract = {OBJECTIVE: To investigate the infection and genotypes of Wolbachia in common mosquito species in Henan Province, so as to provide insights into management of mosquito-borne diseases.

METHODS: Aedes, Culex and Anopheles samples were collected from cowsheds, sheepfolds and human houses in Puyang, Nanyang City and Xuchang cities of Henan Province from July to September, 2022, and the infection of Wolbachia was detected. The 16S rDNA and wsp genes of Wolbachia were amplified and sequenced. Sequence alignment was performed using the BLAST software, and the obtained 16S rDNA gene sequence was compared with the sequence of the 16S rDNA gene in GenBank database. In addition, the phylogenetic trees were created based on 16S rDNA and wsp gene sequences using the software MEGA 11.0.

RESULTS: A total 506 female adult mosquitoes were collected from three sampling sites in Nanyang, Xuchang City and Puyang cities from July to September, 2022. The overall detection of Wolbachia was 45.1% (228/506) in mosquitoes, with a higher detection rate in A. albopictus than in Cx. pipiens pallens [97.9% (143/146) vs. 50.6% (85/168); χ[2] = 88.064, P < 0.01]. The detection of Wolbachia in Cx. pipiens pallens was higher in Xuchang City (96.8%, 62/64) than in Nanyang (15.6%, 7/45) and Puyang cities (27.1%, 16/59) (χ[2] = 89.950, P < 0.01). The homologies of obtained Wolbachia 16S rDNA and wsp gene sequences were 95.3% to 100.0% and 81.7% to 99.8%. Phylogenetic analysis based on wsp gene sequences showed Wolbachia supergroups A and B in mosquito samples, with wAlbA and wMors strains in supergroup A and wPip and wAlbB strains in supergroup B. Wolbachia strain wAlbB infection was detected in A. albopictus in Puyang and Nanyang Cities, while Wolbachia strain wPip infection was identified in A. albopictus in Xuchang City. Wolbachia strain wAlbA infection was detected in Cx. pipiens pallens sampled from three cities, and one Cx. pipiens pallens was found to be infected with Wolbachia strain wMors in Nanyang City.

CONCLUSIONS: Wolbachia infection is commonly prevalent in Ae. albopictus and Cx. pipiens pallens from Henan Province, and Wolbachia strains wAlbB and wAlbA are predominant in Ae. albopictus, while wPip strain is predominant in Cx. pipiens pallens. This is the first report to present Wolbachia wMors strain infection in Cx. pipiens pallens in Henan Province.}, } @article {pmid37926385, year = {2024}, author = {Rodríguez-Rojas, JJ and Lozano-Sardaneta, YN and Fernández-Salas, I and Sánchez-Casas, RM and Becker, I}, title = {Species diversity, barcode, detection of pathogens and blood meal pattern in Phlebotominae (Diptera: Psychodidae) from northeastern Mexico.}, journal = {Acta tropica}, volume = {249}, number = {}, pages = {107064}, doi = {10.1016/j.actatropica.2023.107064}, pmid = {37926385}, issn = {1873-6254}, mesh = {Animals ; Male ; Female ; Humans ; *Psychodidae/parasitology ; Mexico ; Insect Vectors/parasitology ; *Leishmania/genetics ; *Leishmaniasis ; Feeding Behavior ; }, abstract = {More than 90 species of phlebotomines are vectors of parasites, bacteria, and viruses, which cause disease in animals and humans. Therefore, their study is necessary to establish prevention and control strategies. Mexico is an endemic country for leishmaniasis, mostly in the center and southern regions of the country, yet only few studies have been conducted in the northern part of the country. The present study aims to: (a) assess the alpha diversity of Phlebotominae in an annual cycle, (b) to correlate climatic variables with abundance, (c) to generate barcodes of these insects as part of the integrative taxonomy, and (d) to detect Leishmania, Wolbachia and blood sources in an area close to where a case of autochthonous leishmaniasis has been detected in Nuevo Leon, Mexico. A systematic sampling was conducted during three consecutive nights from 17:00 to 22:00 h., placing Shannon traps, CDC traps with incandescent light, and BG Sentinel 2 + BG Lure traps. A total catch effort of 660 nights/traps/hours was achieved, in which a total number of 707 phlebotomines (58% female and 42% male) of six species were collected and identified. The most abundant species were Psathyromyia cratifer (57%) and Psathyromyia shannoni sensu stricto (26%). The highest abundance (72%; 507/707) was collected during March, April and May 2021. Barcodes were generated for four species of phlebotomines, which represent new records for Mexico. For the molecular detection of microorganisms, 302 specimens were analyzed, although no specimens were positive for Leishmania spp. Wolbachia strains were detected in phlebotomines with an infection rate of 1.32% (4/302) and found in Pa. cratifer and Lu. cruciata. Likewise, human DNA was identified in female Lu. cruciata and Pa. cratifer phlebotomines. These findings indicate the presence of potential vector species of the parasite Leishmania spp. This result shows the need for further entomological surveillance to elucidate the transmission mechanisms in these northern areas of the country.}, } @article {pmid37926187, year = {2023}, author = {Badger, JH and Giordano, R and Zimin, A and Wappel, R and Eskipehlivan, SM and Muller, S and Donthu, R and Soto-Adames, F and Vieira, P and Zasada, I and Goodwin, S}, title = {Direct sequencing of insect symbionts via nanopore adaptive sampling.}, journal = {Current opinion in insect science}, volume = {61}, number = {}, pages = {101135}, doi = {10.1016/j.cois.2023.101135}, pmid = {37926187}, issn = {2214-5753}, abstract = {Insect symbionts can alter their host phenotype and their effects can range from beneficial to pathogenic. Moreover, many insects exhibit co-infections, making their study more challenging. Less than 1% of insect species have high-quality referenced genomes available and fewer still also have their symbionts sequenced. Two methods are commonly used to sequence symbionts: whole-genome sequencing to concomitantly capture the host and bacterial genomes, or isolation of the symbiont's genome before sequencing. These methods are limited when dealing with rare or poorly characterized symbionts. Long-read technology is an important tool to generate high-quality genomes as they can overcome high levels of heterozygosity, repeat content, and transposable elements that confound short-read methods. Oxford Nanopore (ONT) adaptive sampling allows a sequencing instrument to select or reject sequences in real time. We describe a method based on ONT adaptive sampling (subtractive) approach that readily permitted the sequencing of the complete genomes of mitochondria, Buchnera and its plasmids (pLeu, pTrp), and Wolbachia genomes in two aphid species, Aphis glycines and Pentalonia nigronervosa. Adaptive sampling is able to retrieve organelles such as mitochondria and symbionts that have high representation in their hosts such as Buchnera and Wolbachia, but is less successful at retrieving symbionts in low concentrations.}, } @article {pmid37923779, year = {2023}, author = {Flores, GAM and Lopez, RP and Cerrudo, CS and Perotti, MA and Consolo, VF and Berón, CM}, title = {Wolbachia dominance influences the Culex quinquefasciatus microbiota.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {18980}, pmid = {37923779}, issn = {2045-2322}, mesh = {Animals ; *Aedes/genetics ; Bacteria/genetics ; *Culex/genetics ; *Microbiota ; Mosquito Vectors/microbiology ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {Microorganisms present in mosquitoes and their interactions are key factors affecting insect development. Among them, Wolbachia is closely associated with the host and affects several fitness parameters. In this study, the bacterial and fungal microbiota from two laboratory Culex quinquefasciatus isolines (wild type and tetracycline-cured) were characterized by metagenome amplicon sequencing of the ITS2 and 16S rRNA genes at different developmental stages and feeding conditions. We identified 572 bacterial and 61 fungal OTUs. Both isolines presented variable bacterial communities and different trends in the distribution of diversity among the groups. The lowest bacterial richness was detected in sugar-fed adults of the cured isoline, whereas fungal richness was highly reduced in blood-fed mosquitoes. Beta diversity analysis indicated that isolines are an important factor in the differentiation of mosquito bacterial communities. Considering composition, Penicillium was the dominant fungal genus, whereas Wolbachia dominance was inversely related to that of Enterobacteria (mainly Thorsellia and Serratia). This study provides a more complete overview of the mosquito microbiome, emphasizing specific highly abundant components that should be considered in microorganism manipulation approaches to control vector-borne diseases.}, } @article {pmid37921460, year = {2023}, author = {Zhang, Y and Liu, S and Huang, X-y and Zi, H-b and Gao, T and Ji, R-j and Sheng, J and Zhi, D and Zhang, Y-l and Gong, C-m and Yang, Y-q}, title = {Altitude as a key environmental factor shaping microbial communities of tea green leafhoppers (Matsumurasca onukii).}, journal = {Microbiology spectrum}, volume = {11}, number = {6}, pages = {e0100923}, pmid = {37921460}, issn = {2165-0497}, support = {No.2021YFD1601105//MOST | National Key Research and Development Program of China (NKPs)/ ; No.32172635//MOST | National Natural Science Foundation of China (NSFC)/ ; }, mesh = {Animals ; *Hemiptera ; Altitude ; Tea ; }, abstract = {Host-associated microbial communities play an important role in the fitness of insect hosts. However, the factors shaping microbial communities in wild populations, including environmental factors and interactions among microbial species, remain largely unknown. The tea green leafhopper has a wide geographical distribution and is highly adaptable, providing a suitable model for studying the effect of ecological drivers on microbiomes. This is the first large-scale culture-independent study investigating the microbial communities of M. onukii sampled from different locations. Altitude as a key environmental factor may have shaped microbial communities of M. onukii by affecting the relative abundance of endosymbionts, especially Wolbachia. The results of this study, therefore, offer not only an in-depth view of the microbial diversity of this species but also an insight into the influence of environmental factors.}, } @article {pmid37914998, year = {2023}, author = {Liu, W and Xia, X and Hoffmann, AA and Ding, Y and Fang, JC and Yu, H}, title = {Evolution of Wolbachia reproductive and nutritional mutualism: insights from the genomes of two novel strains that double infect the pollinator of dioecious Ficus hirta.}, journal = {BMC genomics}, volume = {24}, number = {1}, pages = {657}, pmid = {37914998}, issn = {1471-2164}, support = {2021A1515110981//Guangdong Basic and Applied Basic Research Foundation/ ; 2022ZB773//Jiangsu Funding Program for Excellent Postdoctoral Talent/ ; 2022VBA0002//The Chinese Academy of Sciences PIFI Fellowship for Visiting Scientists/ ; 2023YFE0100540//National Key R & D Program of China/ ; 202206010058//Guangzhou Collaborative Innovation Center on Science-tech of Ecology and Landscape/ ; }, mesh = {*Ficus/genetics ; *Wolbachia/genetics ; Biotin/genetics ; Symbiosis/genetics ; Phylogeny ; DNA Transposable Elements/genetics ; Multilocus Sequence Typing ; Prophages/genetics ; Reproduction ; }, abstract = {Wolbachia is a genus of maternally inherited endosymbionts that can affect reproduction of their hosts and influence metabolic processes. The pollinator, Valisia javana, is common in the male syconium of the dioecious fig Ficus hirta. Based on a high-quality chromosome-level V. javana genome with PacBio long-read and Illumina short-read sequencing, we discovered a sizeable proportion of Wolbachia sequences and used these to assemble two novel Wolbachia strains belonging to supergroup A. We explored its phylogenetic relationship with described Wolbachia strains based on MLST sequences and the possibility of induction of CI (cytoplasmic incompatibility) in this strain by examining the presence of cif genes known to be responsible for CI in other insects. We also identified mobile genetic elements including prophages and insertion sequences, genes related to biotin synthesis and metabolism. A total of two prophages and 256 insertion sequences were found. The prophage WOjav1 is cryptic (structure incomplete) and WOjav2 is relatively intact. IS5 is the dominant transposon family. At least three pairs of type I cif genes with three copies were found which may cause strong CI although this needs experimental verification; we also considered possible nutritional effects of the Wolbachia by identifying genes related to biotin production, absorption and metabolism. This study provides a resource for further studies of Wolbachia-pollinator-host plant interactions.}, } @article {pmid37907954, year = {2023}, author = {Pfarr, KM and Krome, AK and Al-Obaidi, I and Batchelor, H and Vaillant, M and Hoerauf, A and Opoku, NO and Kuesel, AC}, title = {The pipeline for drugs for control and elimination of neglected tropical diseases: 2. Oral anti-infective drugs and drug combinations for off-label use.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {394}, pmid = {37907954}, issn = {1756-3305}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Humans ; *Ivermectin/therapeutic use ; Rifampin ; Doxycycline ; Fluconazole ; Off-Label Use ; *Anti-Infective Agents/therapeutic use ; Drug Combinations ; Neglected Diseases/drug therapy/prevention & control ; }, abstract = {In its 'Road map for neglected tropical diseases 2021-2030', the World Health Organization outlined its targets for control and elimination of neglected tropical diseases (NTDs) and research needed to achieve them. For many NTDs, this includes research for new treatment options for case management and/or preventive chemotherapy. Our review of small-molecule anti-infective drugs recently approved by a stringent regulatory authority (SRA) or in at least Phase 2 clinical development for regulatory approval showed that this pipeline cannot deliver all new treatments needed. WHO guidelines and country policies show that drugs may be recommended for control and elimination for NTDs for which they are not SRA approved (i.e. for 'off-label' use) if efficacy and safety data for the relevant NTD are considered sufficient by WHO and country authorities. Here, we are providing an overview of clinical research in the past 10 years evaluating the anti-infective efficacy of oral small-molecule drugs for NTD(s) for which they are neither SRA approved, nor included in current WHO strategies nor, considering the research sponsors, likely to be registered with a SRA for that NTD, if found to be effective and safe. No such research has been done for yaws, guinea worm, Trypanosoma brucei gambiense human African trypanosomiasis (HAT), rabies, trachoma, visceral leishmaniasis, mycetoma, T. b. rhodesiense HAT, echinococcosis, taeniasis/cysticercosis or scabies. Oral drugs evaluated include sparfloxacin and acedapsone for leprosy; rifampicin, rifapentin and moxifloxacin for onchocerciasis; imatinib and levamisole for loiasis; itraconazole, fluconazole, ketoconazole, posaconazole, ravuconazole and disulfiram for Chagas disease, doxycycline and rifampicin for lymphatic filariasis; arterolane, piperaquine, artesunate, artemether, lumefantrine and mefloquine for schistosomiasis; ivermectin, tribendimidine, pyrantel, oxantel and nitazoxanide for soil-transmitted helminths including strongyloidiasis; chloroquine, ivermectin, balapiravir, ribavirin, celgosivir, UV-4B, ivermectin and doxycycline for dengue; streptomycin, amoxicillin, clavulanate for Buruli ulcer; fluconazole and isavuconazonium for mycoses; clarithromycin and dapsone for cutaneous leishmaniasis; and tribendimidine, albendazole, mebendazole and nitazoxanide for foodborne trematodiasis. Additional paths to identification of new treatment options are needed. One promising path is exploitation of the worldwide experience with 'off-label' treatment of diseases with insufficient treatment options as pursued by the 'CURE ID' initiative.}, } @article {pmid37901801, year = {2023}, author = {Minwuyelet, A and Petronio, GP and Yewhalaw, D and Sciarretta, A and Magnifico, I and Nicolosi, D and Di Marco, R and Atenafu, G}, title = {Symbiotic Wolbachia in mosquitoes and its role in reducing the transmission of mosquito-borne diseases: updates and prospects.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1267832}, pmid = {37901801}, issn = {1664-302X}, abstract = {Mosquito-borne diseases such as malaria, dengue fever, West Nile virus, chikungunya, Zika fever, and filariasis have the greatest health and economic impact. These mosquito-borne diseases are a major cause of morbidity and mortality in tropical and sub-tropical areas. Due to the lack of effective vector containment strategies, the prevalence and severity of these diseases are increasing in endemic regions. Nowadays, mosquito infection by the endosymbiotic Wolbachia represents a promising new bio-control strategy. Wild-infected mosquitoes had been developing cytoplasmic incompatibility (CI), phenotypic alterations, and nutrition competition with pathogens. These reduce adult vector lifespan, interfere with reproduction, inhibit other pathogen growth in the vector, and increase insecticide susceptibility of the vector. Wild, uninfected mosquitoes can also establish stable infections through trans-infection and have the advantage of adaptability through pathogen defense, thereby selectively infecting uninfected mosquitoes and spreading to the entire population. This review aimed to evaluate the role of the Wolbachia symbiont with the mosquitoes (Aedes, Anopheles, and Culex) in reducing mosquito-borne diseases. Global databases such as PubMed, Web of Sciences, Scopus, and pro-Quest were accessed to search for potentially relevant articles. We used keywords: Wolbachia, Anopheles, Aedes, Culex, and mosquito were used alone or in combination during the literature search. Data were extracted from 56 articles' texts, figures, and tables of the included article.}, } @article {pmid37891252, year = {2023}, author = {Lenharo, M}, title = {Dengue rates drop after release of modified mosquitoes in Colombia.}, journal = {Nature}, volume = {623}, number = {7986}, pages = {235-236}, pmid = {37891252}, issn = {1476-4687}, mesh = {Animals ; Humans ; *Aedes/microbiology/virology ; Colombia/epidemiology ; *Dengue/epidemiology/prevention & control/transmission/virology ; *Dengue Virus ; *Wolbachia ; *Biological Control Agents ; }, } @article {pmid37887819, year = {2023}, author = {Xoconostle-Cázares, B and Ramírez-Pool, JA and Núñez-Muñoz, LA and Calderón-Pérez, B and Vargas-Hernández, BY and Bujanos-Muñiz, R and Ruiz-Medrano, R}, title = {The Characterization of Melanaphis sacchari Microbiota and Antibiotic Treatment Effect on Insects.}, journal = {Insects}, volume = {14}, number = {10}, pages = {}, pmid = {37887819}, issn = {2075-4450}, support = {grant CF-2023-G-731//Consejo Nacional de Humanidades Ciencias y Tecnologias/ ; }, abstract = {Insects are under constant selective pressure, which has resulted in adaptations to novel niches such as crops. This is the case of the pest Melanaphis sacchari, the sugarcane aphid, native to Africa and currently spreading worldwide. The aphid undergoes successful parthenogenesis, causing important damage to a variety of crops and leading to important economic losses for farmers. A natural M. sacchari population grown in sorghum was studied to identify its microbiome through the sequencing of its 16S rDNA metagenome. A high proportion of Proteobacteria, followed by Firmicutes, Bacteroidetes, and Actinobacteria, was observed. We also detected Wolbachia, which correlates with the asexual reproduction of its host. M. sacchari was challenged in a bioassay with the antibiotics oxytetracycline and streptomycin, resulting in a dose-dependent decay of its survival rate. The possibility of controlling this pest by altering its microbiota is proposed.}, } @article {pmid37887804, year = {2023}, author = {Liu, Y and Liu, J and Zhang, X and Yun, Y}, title = {Diversity of Bacteria Associated with Guts and Gonads in Three Spider Species and Potential Transmission Pathways of Microbes within the Same Spider Host.}, journal = {Insects}, volume = {14}, number = {10}, pages = {}, pmid = {37887804}, issn = {2075-4450}, support = {2018YFA0901101//The National Key Technology R&D Program of China/ ; 2016AHB003//Key Scientific and Technological Projects of Hubei/ ; }, abstract = {Microbial symbiosis plays a crucial role in the ecological and evolutionary processes of animals. It is well known that spiders, with their unique and diverse predatory adaptations, assume an indispensable role in maintaining ecological balance and the food chain. However, our current understanding of spider microbiomes remains relatively limited. The gut microbiota and gonad microbiota of spiders can both potentially influence their physiology, ecology, and behavior, including aspects such as digestion, immunity, reproductive health, and reproductive behavior. In the current study, based on high-throughput sequencing of the 16S rRNA V3 and V4 regions, we detected the gut and gonad microbiota communities of three spider species captured from the same habitat, namely, Eriovixia cavaleriei, Larinioides cornutus, and Pardosa pseudoannulata. In these three species, we observed that, at the phylum level classification, the gut and gonad of E. cavaleriei are primarily composed of Proteobacteria, while those of L. cornutus and P. pseudoannulata are primarily composed of Firmicutes. At the genus level of classification, we identified 372 and 360 genera from the gut and gonad bacterial communities. It is noteworthy that the gut and gonad bacterial flora of E. cavaleriei and L. cornutus were dominated by Wolbachia and Spiroplasma. Results show that there were no differences in microbial communities between females and males of the same spider species. Furthermore, there is similarity between the gut and ovary microbial communities of female spiders, implying a potential avenue for microbial transmission between the gut and gonad within female spiders. By comprehensively studying these two microbial communities, we can establish the theoretical foundation for exploring the relationship between gut and gonad microbiota and their host, as well as the mechanisms through which microbes exert their effects.}, } @article {pmid37882523, year = {2023}, author = {Głowska, E and Gerth, M}, title = {Draft genome sequence of a Wolbachia endosymbiont from Syringophilopsis turdi (Fritsch, 1958) (Acari, Syringophilidae).}, journal = {Microbiology resource announcements}, volume = {12}, number = {11}, pages = {e0060523}, pmid = {37882523}, issn = {2576-098X}, support = {UMO-2015/19/D/NZ8/00191//Narodowe Centrum Nauki (NCN)/ ; GE 2824/1-1//Deutsche Forschungsgemeinschaft (DFG)/ ; }, abstract = {We present the draft genome of a Wolbachia endosymbiont from quill mites. This is the first representative of a recently discovered distinct Wolbachia lineage (supergroup P). We hope the genome will be a useful resource for comparative evolutionary and genomic studies across the globally distributed symbiont Wolbachia.}, } @article {pmid37874788, year = {2023}, author = {Russell, SL and Castillo, JR and Sullivan, WT}, title = {Wolbachia endosymbionts manipulate the self-renewal and differentiation of germline stem cells to reinforce fertility of their fruit fly host.}, journal = {PLoS biology}, volume = {21}, number = {10}, pages = {e3002335}, pmid = {37874788}, issn = {1545-7885}, support = {K99 GM135583/GM/NIGMS NIH HHS/United States ; R00 GM135583/GM/NIGMS NIH HHS/United States ; R35 GM139595/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Wolbachia ; Drosophila/metabolism ; Fertility ; Cell Differentiation ; Germ Cells/metabolism ; Stem Cells/metabolism ; RNA, Messenger/metabolism ; Drosophila melanogaster/genetics ; RNA-Binding Proteins/genetics ; *Drosophila Proteins/metabolism ; }, abstract = {The alphaproteobacterium Wolbachia pipientis infects arthropod and nematode species worldwide, making it a key target for host biological control. Wolbachia-driven host reproductive manipulations, such as cytoplasmic incompatibility (CI), are credited for catapulting these intracellular bacteria to high frequencies in host populations. Positive, perhaps mutualistic, reproductive manipulations also increase infection frequencies, but are not well understood. Here, we identify molecular and cellular mechanisms by which Wolbachia influences the molecularly distinct processes of germline stem cell (GSC) self-renewal and differentiation. We demonstrate that wMel infection rescues the fertility of flies lacking the translational regulator mei-P26 and is sufficient to sustain infertile homozygous mei-P26-knockdown stocks indefinitely. Cytology revealed that wMel mitigates the impact of mei-P26 loss through restoring proper pMad, Bam, Sxl, and Orb expression. In Oregon R files with wild-type fertility, wMel infection elevates lifetime egg hatch rates. Exploring these phenotypes through dual-RNAseq quantification of eukaryotic and bacterial transcripts revealed that wMel infection rescues and offsets many gene expression changes induced by mei-P26 loss at the mRNA level. Overall, we show that wMel infection beneficially reinforces host fertility at mRNA, protein, and phenotypic levels, and these mechanisms may promote the emergence of mutualism and the breakdown of host reproductive manipulations.}, } @article {pmid37873081, year = {2023}, author = {Perlmutter, JI and Atadurdyyeva, A and Schedl, ME and Unckless, RL}, title = {Wolbachia enhances the survival of Drosophila infected with fungal pathogens.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {37873081}, abstract = {Wolbachia bacteria of arthropods are at the forefront of basic and translational research on multipartite host-symbiont-pathogen interactions. These microbes are vertically inherited from mother to offspring via the cytoplasm. They are the most widespread endosymbionts on the planet due to their infamous ability to manipulate the reproduction of their hosts to spread themselves in a population, and to provide a variety of fitness benefits to their hosts. Importantly, some strains of Wolbachia can inhibit viral pathogenesis within and between arthropod hosts. Mosquitoes carrying the wMel Wolbachia strain of Drosophila melanogaster have a greatly reduced capacity to spread viruses like dengue and Zika to humans. Therefore, Wolbachia are the basis of several global vector control initiatives. While significant research efforts have focused on viruses, relatively little attention has been given to Wolbachia-fungal interactions despite the ubiquity of fungal entomopathogens in nature. Here, we demonstrate that Wolbachia increase the longevity of their Drosophila melanogaster hosts when challenged with a spectrum of yeast and filamentous fungal pathogens. We find that this pattern can vary based on host genotype, sex, and fungal species. Further, Wolbachia correlates with higher fertility and reduced pathogen titers during initial fungal infection, indicating a significant fitness benefit. This study demonstrates Wolbachia's role in diverse fungal pathogen interactions and determines that the phenotype is broad, but with several variables that influence both the presence and strength of the phenotype. These results enhance our knowledge of the strategies Wolbachia uses that likely contribute to such a high global symbiont prevalence.}, } @article {pmid37871129, year = {2023}, author = {Wenzel, M and Aquadro, CF}, title = {Wolbachia infection at least partially rescues the fertility and ovary defects of several new Drosophila melanogaster bag of marbles protein-coding mutants.}, journal = {PLoS genetics}, volume = {19}, number = {10}, pages = {e1011009}, pmid = {37871129}, issn = {1553-7404}, mesh = {Animals ; Female ; Male ; *Drosophila melanogaster/genetics/microbiology ; *Drosophila Proteins/genetics/metabolism ; Fertility/genetics ; Ovary/metabolism ; *Wolbachia/genetics/metabolism ; }, abstract = {The D. melanogaster protein coding gene bag of marbles (bam) plays a key role in early male and female reproduction by forming complexes with partner proteins to promote differentiation in gametogenesis. Like another germline gene, Sex lethal, bam genetically interacts with the endosymbiont Wolbachia, as Wolbachia rescues the reduced fertility of a bam hypomorphic mutant. Here, we explored the specificity of the bam-Wolbachia interaction by generating 22 new bam mutants, with ten mutants displaying fertility defects. Nine of these mutants trend towards rescue by the wMel Wolbachia variant, with eight statistically significant at the fertility and/or cytological level. In some cases, fertility was increased a striking 20-fold. There is no specificity between the rescue and the known binding regions of bam, suggesting wMel does not interact with one singular bam partner to rescue the reproductive phenotype. We further tested if wMel interacts with bam in a non-specific way, by increasing bam transcript levels or acting upstream in germline stem cells. A fertility assessment of a bam RNAi knockdown mutant reveals that wMel rescue is specific to functionally mutant bam alleles and we find no obvious evidence of wMel interaction with germline stem cells in bam mutants.}, } @article {pmid37860089, year = {2023}, author = {Rushidi, MNA and Azhari, MLH and Yaakop, S and Hazmi, IR}, title = {Detection and Characterisation of Endosymbiont Wolbachia (Rickettsiales: Anaplasmataceae) in Elaeidobius kamerunicus (Coleoptera: Curculionoidea), Pollinating Agent of Oil Palm, and Its Relationships between Populations.}, journal = {Tropical life sciences research}, volume = {34}, number = {3}, pages = {95-111}, pmid = {37860089}, issn = {1985-3718}, abstract = {Elaeidobius kamerunicus is the most efficient pollinator of oil palm. Wolbachia is an endosymbiotic bacteria associated with E. kamerunicus that has a potential to affect the fecundity and fitness of the E. kamerunicus. Despite their importance, no studies have been conducted to investigate its prevalence in E. kamerunicus. The objectives of this study were to detect and characterise Wolbachia in E. kamerunicus and determine the phylogenetic relationship of Wolbachia strains that infect E. kamerunicus by using three genetic markers namely Filamenting temperature-sensitive mutant Z (ftsZ), Chaperonin folding protein (groEL), and Citrate Synthase Coding Gene (gltA). DNA was extracted from 210 individuals of E. kamerunicus and the Wolbachia infections were detected using the wsp marker. The infected samples (n = 25, 11.9%) were then sequenced using ftsZ, gltA and groEL markers for strain characterization. In this study, a combination of four markers was used to construct the phylogeny of Wolbachia. Similar topologies were shown in all trees; Neighbour-Joining (NJ), Maximum Parsimony (MP), and Bayesian Inference (BI), which showed the mixing of individuals that harbor Wolbachia between populations. Interestingly, Wolbachia on E. kamerunicus was claded together with the species Drosophila simulans under supergroup B. This is the first report of Wolbachia infecting E. kamerunicus which is very valuable and significant as one of the parameters to evaluate the quality of the E. kamerunicus population for sustaining its function as a great pollinator for oil palm.}, } @article {pmid37859959, year = {2023}, author = {Ridha, MR and Marlinae, L and Zubaidah, T and Fadillah, NA and Widjaja, J and Rosadi, D and Rahayu, N and Ningsih, M and Desimal, I and Sofyandi, A}, title = {Control methods for invasive mosquitoes of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Indonesia.}, journal = {Veterinary world}, volume = {16}, number = {9}, pages = {1952-1963}, pmid = {37859959}, issn = {0972-8988}, abstract = {The two invasive mosquito species in Indonesia are Aedes aegypti and Ae. albopictus. These mosquitoes are a serious nuisance to humans and are also the primary vectors of several foreign pathogens, such as dengue, Zika, and chikungunya viruses. Efforts must be made to reduce the possibility of mosquito bites and the potential for disease transmission. Given the invasion of these two Aedes species, this approach should be considered as part of an integrated strategy to manage them. This review discusses existing and developing control techniques for invasive Ae. aegypti and Ae. albopictus, with an emphasis on those that have been and are being used in Indonesia. Environmental, mechanical, biological (e.g., Bacillus thuringiensis and Wolbachia), and chemical (e.g., insect growth regulators and pyrethroids) approaches are discussed in this review, considering their effectiveness, sustainability, and control methods.}, } @article {pmid37838705, year = {2023}, author = {Sounart, H and Voronin, D and Masarapu, Y and Chung, M and Saarenpää, S and Ghedin, E and Giacomello, S}, title = {Miniature spatial transcriptomics for studying parasite-endosymbiont relationships at the micro scale.}, journal = {Nature communications}, volume = {14}, number = {1}, pages = {6500}, pmid = {37838705}, issn = {2041-1723}, mesh = {Animals ; Female ; Humans ; *Parasites/genetics ; Transcriptome ; Anti-Bacterial Agents/metabolism ; Gene Expression Profiling ; *Communicable Diseases ; *Wolbachia/genetics/metabolism ; Symbiosis/genetics ; }, abstract = {Several important human infectious diseases are caused by microscale-sized parasitic nematodes like filarial worms. Filarial worms have their own spatial tissue organization; to uncover this tissue structure, we need methods that can spatially resolve these miniature specimens. Most filarial worms evolved a mutualistic association with endosymbiotic bacteria Wolbachia. However, the mechanisms underlying the dependency of filarial worms on the fitness of these bacteria remain unknown. As Wolbachia is essential for the development, reproduction, and survival of filarial worms, we spatially explored how Wolbachia interacts with the worm's reproductive system by performing a spatial characterization using Spatial Transcriptomics (ST) across a posterior region containing reproductive tissue and developing embryos of adult female Brugia malayi worms. We provide a proof-of-concept for miniature-ST to explore spatial gene expression patterns in small sample types, demonstrating the method's ability to uncover nuanced tissue region expression patterns, observe the spatial localization of key B. malayi - Wolbachia pathway genes, and co-localize the B. malayi spatial transcriptome in Wolbachia tissue regions, also under antibiotic treatment. We envision our approach will open up new avenues for the study of infectious diseases caused by micro-scale parasitic worms.}, } @article {pmid37830551, year = {2023}, author = {Riparbelli, MG and Pratelli, A and Callaini, G}, title = {Wolbachia Induces Structural Defects Harmful to Drosophila simulans Riverside Spermiogenesis.}, journal = {Cells}, volume = {12}, number = {19}, pages = {}, pmid = {37830551}, issn = {2073-4409}, mesh = {Animals ; Male ; *Drosophila simulans ; Drosophila ; *Wolbachia ; Semen ; Spermatogenesis ; }, abstract = {The relationship between cytoplasmic incompatibility and the obligate intracellular alphaproteobacteria Wolbachia has for a long time been reported. Although the molecular mechanisms responsible for this reproductive alteration are beginning to be understood, the effects of Wolbachia on germ cell structure and dynamics have not yet been fully investigated. We report here that the presence of Wolbachia in infected cysts of elongating spermatids is associated with major structural defects that become more evident in mature sperm. We find mitochondrial defects, an improper axoneme structure, reduced sperm numbers, and individualization failures. The large heterogeneous variety of the ultrastructural defects found in elongating spermatids and mature sperm provide the first cytological evidence for the reduced fertility associated with Wolbachia infection in Drosophila simulans males. The observed abnormalities could be the result of the mechanical stress induced by the high bacteria numbers during the process of spermatid elongation, rather than the result of the released factors affecting the proper morphogenesis of the germ cells. Moreover, high Wolbachia densities in male germ cells may not be appropriate for causing cytoplasmic incompatibility as the bacteria are harmful for spermatid differentiation, leading to abnormal sperm that is unlikely to be functional.}, } @article {pmid37822704, year = {2023}, author = {Pavan, MG and Garcia, GA and David, MR and Maciel-de-Freitas, R}, title = {The double-edged sword effect of expanding Wolbachia deployment in dengue endemic settings.}, journal = {Lancet regional health. Americas}, volume = {27}, number = {}, pages = {100610}, pmid = {37822704}, issn = {2667-193X}, } @article {pmid37819592, year = {2023}, author = {Haghshenas-Gorgabi, N and Poorjavd, N and Khajehali, J and Wybouw, N}, title = {Cardinium symbionts are pervasive in Iranian populations of the spider mite Panonychus ulmi despite inducing an infection cost and no demonstrable reproductive phenotypes when Wolbachia is a symbiotic partner.}, journal = {Experimental & applied acarology}, volume = {91}, number = {3}, pages = {369-380}, pmid = {37819592}, issn = {1572-9702}, mesh = {Female ; Male ; Animals ; *Wolbachia ; *Tetranychidae/genetics ; Iran ; Seeds ; Reproduction ; Symbiosis ; Bacteroidetes ; }, abstract = {Maternally transmitted symbionts such as Cardinium and Wolbachia are widespread in arthropods. Both Cardinium and Wolbachia can cause cytoplasmic incompatibility, a reproductive phenotype that interferes with the development of uninfected eggs that are fertilized by infected sperm. In haplodiploid hosts, these symbionts can also distort sex allocation to facilitate their spread through host populations. Without other fitness effects, symbionts that induce strong reproductive phenotypes tend to spread to high and stable infection frequencies, whereas variants that induce weak reproductive phenotypes are typically associated with intermediate and variable frequencies. To study the spread of Cardinium in a haplodiploid host, we sampled Iranian populations of the economically important spider mite Panonychus ulmi in apple orchards. Within several field populations, we also studied the Wolbachia infection frequencies. All P. ulmi field populations carried a Cardinium infection and exhibited high infection frequencies. In contrast, Wolbachia frequency ranged between ca. 10% and ca. 70% and was only found in co-infected mites. To test whether Cardinium induce reproductive phenotypes in P. ulmi, a Cardinium-cured derived line was generated by antibiotic treatment from a co-infected field population. Genetic crosses indicated that Cardinium do not induce demonstrable levels of cytoplasmic incompatibility and sex allocation distortion in co-infected P. ulmi. However, Cardinium infection was associated with a longer developmental time and reduced total fecundity for co-infected females. We hypothesize that Cardinium spread through P. ulmi populations via uncharacterized fitness effects and that co-infection with Wolbachia might impact these drive mechanisms.}, } @article {pmid37811984, year = {2023}, author = {Edwards, B and Ghedin, E and Voronin, D}, title = {Wolbachia interferes with Zika virus replication by hijacking cholesterol metabolism in mosquito cells.}, journal = {Microbiology spectrum}, volume = {11}, number = {6}, pages = {e0218023}, pmid = {37811984}, issn = {2165-0497}, mesh = {Animals ; Humans ; *Zika Virus ; *Wolbachia ; *Aedes/microbiology ; Mosquito Vectors/microbiology ; *Zika Virus Infection ; *Arboviruses ; Virus Replication ; Cholesterol ; }, abstract = {Arthropod-borne viruses are emerging pathogens that are spread widely by mosquitos. Zika virus is an arbovirus that can infect humans and be transmitted from an infected mother to the fetus, potentially leading to microcephaly in infants. One promising strategy to prevent disease caused by arboviruses is to target the insect vector population. Recent field studies have shown that mosquito populations infected with Wolbachia bacteria suppress arbovirus replication and transmission. Here, we describe how intracellular bacteria redirect resources within their host cells and suppress Zika virus replication at the cellular level. Understanding the mechanism behind Wolbachia-induced interference of arbovirus replication could help advance strategies to control arbovirus pathogens in insect vectors and human populations.}, } @article {pmid37808371, year = {2023}, author = {Sharma, AK and Som, A}, title = {Assigning new supergroups V and W to the Wolbachia diversity.}, journal = {Bioinformation}, volume = {19}, number = {3}, pages = {336-340}, pmid = {37808371}, issn = {0973-2063}, abstract = {Wolbachia are endosymbiotic and alphaproteobacteria that belong to the order Rickettsiales. They are known to infect half of the insect population and cause host manipulation, and have been categorized into 19 monophyletic lineages called supergroups. Recently, two strains, wCfeJ and wCfeT were isolated from cat fleas (Ctenocephalides felis), but their supergroup relationships were not assigned. In this article, we have attempted to classify these two novel strains and establish their evolutionary lineage (i.e., supergroup designation). For this we performed 16S rRNA similarity analysis and reconstructed 16S rRNA phylogeny of 52 Wolbachia strains (including two novel strains) belong to 19 supergroups. We also performed average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) studies to measure genomic similarity between the two novel genomes. The results revealed that 16S rRNA similarity between the two novel strains is 97.94%, which is below the threshold value of 98.6% and phylogeny shows that they are placed at the two different positions (i.e., showing distinct evolutionary lineages). Further, genomic similarity analysis revealed that the novel genomes have ANI and dDDH values 79% and 22.4% respectively, which were below the threshold value of ANI (95%) and dDDH (70%). These results suggested that the novel strains neither shared a species boundary between them nor with any other previously identified supergroups, which designate them as two new supergroups, namely supergroup V (strain wCfeJ) and supergroup W (strain wCfeT).}, } @article {pmid37808301, year = {2023}, author = {Cui, X and Liu, Y and Zhang, J and Hu, P and Zheng, Z and Deng, X and Xu, M}, title = {Variation of endosymbiont and citrus tristeza virus (CTV) titers in the Huanglongbing insect vector, Diaphorina citri, on CTV-infected plants.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1236731}, pmid = {37808301}, issn = {1664-302X}, abstract = {"Candidatus Liberibacter asiaticus" (CLas) is a notorious agent that causes Citrus Huanglongbing (HLB), which is transmitted by Diaphorina citri (D. citri). We recently found that the acquisition and transmission of CLas by D. citri was facilitated by Citrus tristeza virus (CTV), a widely distributed virus in the field. In this study, we further studied whether different CTV strains manipulate the host preference of D. citri, and whether endosymbionts variation is related to CTV strains in D. citri. The results showed that the non-viruliferous D. citri preferred to select the shoots infected with CTV, without strain differences was observed in the selection. However, the viruliferous D. citri prefered to select the mixed strain that is similar to the field's. Furthermore, D. citri effectively acquired the CTV within 2-12 h depending on the strains of the virus. The persistence period of CTV in D. citri was longer than 24 days, without reduction of the CTV titers being observed. These results provide a foundation for understanding the transmission mode of D. citri on CTV. During the process of CTV acquisition and persistence, the titers of main endosymbionts in D. citri showed similar variation trend, but their relative titers were different at different time points. The titers of the "Candidatus Profftella armatura" and CTV tended to be positively correlated, and the titers of Wolbachia and "Candidatus Carsonella ruddii" were mostly negatively related with titers of CT31. These results showed the relationship among D. citri, endosymbionts, and CTV and provided useful information for further research on the interactions between D. citri and CLas, which may benefit the development of approaches for the prevention of CLas transmission and control of citrus HLB.}, } @article {pmid37808105, year = {2023}, author = {Pikula, J and Piacek, V and Bandouchova, H and Bartlova, M and Bednarikova, S and Burianova, R and Danek, O and Jedlicka, P and Masova, S and Nemcova, M and Seidlova, V and Zukalova, K and Zukal, J}, title = {Case report: Filarial infection of a parti-coloured bat: Litomosa sp. adult worms in abdominal cavity and microfilariae in bat semen.}, journal = {Frontiers in veterinary science}, volume = {10}, number = {}, pages = {1284025}, pmid = {37808105}, issn = {2297-1769}, abstract = {BACKGROUND: Filarial infections have been understudied in bats. Likewise, little is known about pathogens associated with the reproductive system in chiropterans. While semen quality is critical for reproductive success, semen-borne pathogens may contribute to reproductive failure.

METHODS: For the first time we performed electroejaculation and used computer-assisted semen analysis to provide baseline data on semen quality in a parti-coloured bat (Vespertilio murinus).

RESULTS: The semen quality values measured in the V. murinus male appeared high (semen concentration = 305.4 × 10[6]/mL; progressive and motile sperm = 46.58 and 60.27%, respectively). As an incidental finding, however, microfilariae were observed in the bat semen examined. At necropsy, eight adult filarial worms, later genetically identified as Litomosa sp., were found in the peritoneal cavity, close to the stomach, of the same particoloured bat male dying as a result of dysmicrobia and haemorrhagic gastroenteritis in a wildlife rescue centre. Histopathology revealed microfilariae in the testicular connective tissue and the epidydimal connective and fat tissues. A PCR assay targeting cytochrome c oxidase subunit 1 confirmed that adult worms from the peritoneal cavity and testicular microfilariae were of the same filarial species. Mildly engorged argasid mite larvae attached to the bat skin proved negative for filarial DNA and the adult filarial worms proved negative for endosymbiont Wolbachia.

CONCLUSION: While the standard filarial life cycle pattern involves a vertebrate definitive host and an invertebrate vector, represented by a blood-sucking ectoparasite, our finding suggests that microfilariae of this nematode species may also be semen-borne, with transmission intensity promoted by the polygynous mating system of vespertilionid bats in which an infected male mates with many females during the autumn swarming. Presence of microfilariae may be expected to decrease semen quality and transmission via this route may challenge the success of reproductive events in females after mating. Further investigation will be necessary to better understand the bat-parasite interaction and the life cycle of this filarial worm.}, } @article {pmid37804520, year = {2023}, author = {Sri-Pathmanathan, C and Bao, H and Diluka, PAE and Mee, A and Andari, B and Saunders, E and Wijegunawardana, A and Weerasinghe, I and Hetti, NP and Samaraweera, S and Tang, W and Tucker, JD}, title = {Enhancing Community Participation in Dengue Control Through Digital Crowdsourcing: An Analysis of a World Mosquito Program Digital Open Call in Sri Lanka.}, journal = {The Journal of infectious diseases}, volume = {228}, number = {11}, pages = {1482-1490}, doi = {10.1093/infdis/jiad439}, pmid = {37804520}, issn = {1537-6613}, support = {//World Mosquito Program/ ; }, mesh = {Animals ; Humans ; Adolescent ; Young Adult ; Adult ; *Culicidae ; *Crowdsourcing/methods ; Sri Lanka/epidemiology ; Community Participation ; *Dengue/epidemiology/prevention & control ; Mosquito Control ; }, abstract = {BACKGROUND: Two crowdsourcing open calls were created to enhance community engagement in dengue control in Sri Lanka. We analyzed the process and outcomes of these digital crowdsourcing open calls.

METHODS: We used standard World Health Organization methods to organize the open calls, which used exclusively digital methods because of coronavirus disease 2019 (COVID-19). We collected and analyzed sociodemographic information and digital engagement metrics from each submission. Submissions in the form of textual data describing community-led strategies for mosquito release were coded using grounded theory.

RESULTS: The open calls received 73 submissions. Most people who submitted ideas spoke English, lived in Sri Lanka, and were 18 to 34 years old. The total Facebook reach was initially limited (16 161 impressions), prompting expansion to a global campaign, which reached 346 810 impressions over 14 days. Diverse strategies for the distribution of Wolbachia-infected mosquito boxes were identified, including leveraging traditional festivals, schools, and community networks. Fifteen submissions (21%) suggested the use of digital tools for monitoring and evaluation, sharing instructions, or creating networks. Thirteen submissions (18%) focused on social and economic incentives to prompt community engagement and catalyze community-led distribution.

CONCLUSIONS: Our project demonstrates that digital crowdsourcing open calls are an effective way to solicit creative and innovative ideas in a resource-limited setting.}, } @article {pmid37803282, year = {2023}, author = {Somia, ES and Ullah, I and Alyahya, HS and Mahyoub, JA}, title = {Identification of Wolbachia new strains from Aedes aegypti mosquitoes, the vector of dengue fever in Jeddah Province.}, journal = {BMC microbiology}, volume = {23}, number = {1}, pages = {287}, pmid = {37803282}, issn = {1471-2180}, mesh = {Male ; Animals ; Humans ; *Aedes ; *Wolbachia/genetics ; Phylogeny ; Multilocus Sequence Typing ; Mosquito Vectors ; *Dengue/epidemiology ; }, abstract = {Wolbachia are endosymbiotic bacteria found within many arthropods, including insects. A variety of benefits are provided by these bacteria to human and insect hosts, including protection from viruses and parasites and the ability to kill males. In this study, Wolbachia was identified in Aedes aegypti present in Jeddah, Saudi Arabia. A population of mosquitoes was collected from eight different areas, processed, and tested for Wolbachia using 16 S rRNA specific to Wolbachia bacteria and Wolbachia surface protein (wsp) under optimized PCR conditions. In five ecologically diverse sites to determine Wolbachia prevalence, we identified eleven diverse novel resident Wolbachia strains within Ae. Aegypti for the first time in Jeddah, Saudi Arabia. Future studies to evaluate the possible use of Wolbachia as a control agent in Aedes sp. in Saudi Arabia are necessary. Wolbachia prevalence rates and strain characterization through Sanger sequencing with multilocus sequence typing (MLST) and phylogenetic analysis revealed significant diversity. In developing biocontrol strategies, it is beneficial to consider the implications of resident Wolbachia strains.}, } @article {pmid37799509, year = {2023}, author = {Mercant Osuna, A and Gidley, A and Mayi, MPA and Bamou, R and Dhokiya, V and Antonio-Nkondjio, C and Jeffries, CL and Walker, T}, title = {Diverse novel Wolbachia bacteria strains and genera-specific co-infections with Asaia bacteria in Culicine mosquitoes from ecologically diverse regions of Cameroon.}, journal = {Wellcome open research}, volume = {8}, number = {}, pages = {267}, pmid = {37799509}, issn = {2398-502X}, abstract = {Background: The endosymbiotic bacterium Wolbachia infects numerous species of insects and Wolbachia transinfection of Aedes mosquito species is now being used for biocontrol programs as Wolbachia strains can both inhibit arboviruses and invade wild mosquito populations. The discovery of novel, resident Wolbachia strains in mosquito species warrants further investigation as potential candidate strains for biocontrol strategies. Methods: We obtained mosquito specimens from diverse Culicine mosquitoes from Cameroon including ecologically diverse locations in the Central and West Regions. Wolbachia prevalence rates were assessed in addition to the environmentally acquired bacterial species Asaia in major Culicine genera. PCR-based methods were also used with phylogenetic analysis to confirm identities of host mosquito species and Wolbachia strains were classified using multi-locus sequence typing (MLST). Results: We report high Wolbachia prevalence rates for Culicine species, including in a large cohort of Aedes africanus collected from west Cameroon in which 100% of mono-specific pools were infected. Furthermore, co-infections with Asaia bacteria were observed across multiple genera, demonstrating that these two bacteria can co-exist in wild mosquito populations. Wolbachia strain MLST and phylogenetic analysis provided evidence for diverse Wolbachia strains in 13 different mosquito species across seven different genera. Full or partial MLST profiles were generated from resident Wolbachia strains in six Culex species (quinquefasciatus, watti, cinerus, nigripalpus, perexiguus and rima), two Aedes species (africanus and denderensis) and in Mansonia uniformis, Catageiomyia argenteopunctata, Lutzia tigripes, Eretmapodites chrysogaster and Uranotaenia bilineata. Conclusions: Our study provides further evidence that Wolbachia is widespread within wild mosquito populations of diverse Culicine species and provides further candidate strains that could be investigated as future options for Wolbachia-based biocontrol to inhibit arbovirus transmission.}, } @article {pmid37794084, year = {2023}, author = {Hettiarachchi, A and Cnockaert, M and Joossens, M and Gekière, A and Meeus, I and Vereecken, NJ and Michez, D and Smagghe, G and Vandamme, P}, title = {The wild solitary bees Andrena vaga, Anthophora plumipes, Colletes cunicularius, and Osmia cornuta microbiota are host specific and dominated by endosymbionts and environmental microorganisms.}, journal = {Microbial ecology}, volume = {86}, number = {4}, pages = {3013-3026}, pmid = {37794084}, issn = {1432-184X}, mesh = {Bees ; Animals ; RNA, Ribosomal, 16S/genetics ; *Microbiota ; *Mycobiome ; Bacteria ; *Spiroplasma ; }, abstract = {We characterized the microbial communities of the crop, midgut, hindgut, and ovaries of the wild solitary bees Andrena vaga, Anthophora plumipes, Colletes cunicularius, and Osmia cornuta through 16S rRNA gene and ITS2 amplicon sequencing and a large-scale isolation campaign. The bacterial communities of these bees were dominated by endosymbionts of the genera Wolbachia and Spiroplasma. Bacterial and yeast genera representing the remaining predominant taxa were linked to an environmental origin. While only a single sampling site was examined for Andrena vaga, Anthophora plumipes, and Colletes cunicularius, and two sampling sites for Osmia cornuta, the microbiota appeared to be host specific: bacterial, but not fungal, communities generally differed between the analyzed bee species, gut compartments and ovaries. This may suggest a selective process determined by floral and host traits. Many of the gut symbionts identified in the present study are characterized by metabolic versatility. Whether they exert similar functionalities within the bee gut and thus functional redundancy remains to be elucidated.}, } @article {pmid37782672, year = {2023}, author = {Boehm, EC and Jaeger, AS and Ries, HJ and Castañeda, D and Weiler, AM and Valencia, CC and Weger-Lucarelli, J and Ebel, GD and O'Connor, SL and Friedrich, TC and Zamanian, M and Aliota, MT}, title = {Wolbachia-mediated resistance to Zika virus infection in Aedes aegypti is dominated by diverse transcriptional regulation and weak evolutionary pressures.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {10}, pages = {e0011674}, pmid = {37782672}, issn = {1935-2735}, support = {P51 OD011106/OD/NIH HHS/United States ; P51 RR000167/RR/NCRR NIH HHS/United States ; T32 AI083196/AI/NIAID NIH HHS/United States ; R01 AI132563/AI/NIAID NIH HHS/United States ; R21 AI131454/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Zika Virus/genetics ; *Zika Virus Infection ; *Aedes/physiology ; *Wolbachia/physiology ; *Dengue Virus/physiology ; Mosquito Vectors ; }, abstract = {A promising candidate for arbovirus control and prevention relies on replacing arbovirus-susceptible Aedes aegypti populations with mosquitoes that have been colonized by the intracellular bacterium Wolbachia and thus have a reduced capacity to transmit arboviruses. This reduced capacity to transmit arboviruses is mediated through a phenomenon referred to as pathogen blocking. Pathogen blocking has primarily been proposed as a tool to control dengue virus (DENV) transmission, however it works against a range of viruses, including Zika virus (ZIKV). Despite years of research, the molecular mechanisms underlying pathogen blocking still need to be better understood. Here, we used RNA-seq to characterize mosquito gene transcription dynamics in Ae. aegypti infected with the wMel strain of Wolbachia that are being released by the World Mosquito Program in Medellín, Colombia. Comparative analyses using ZIKV-infected, uninfected tissues, and mosquitoes without Wolbachia revealed that the influence of wMel on mosquito gene transcription is multifactorial. Importantly, because Wolbachia limits, but does not completely prevent, replication of ZIKV and other viruses in coinfected mosquitoes, there is a possibility that these viruses could evolve resistance to pathogen blocking. Therefore, to understand the influence of Wolbachia on within-host ZIKV evolution, we characterized the genetic diversity of molecularly barcoded ZIKV virus populations replicating in Wolbachia-infected mosquitoes and found that within-host ZIKV evolution was subject to weak purifying selection and, unexpectedly, loose anatomical bottlenecks in the presence and absence of Wolbachia. Together, these findings suggest that there is no clear transcriptional profile associated with Wolbachia-mediated ZIKV restriction, and that there is no evidence for ZIKV escape from this restriction in our system.}, } @article {pmid37779725, year = {2023}, author = {Guo, Y and Shao, J and Wu, Y and Li, Y}, title = {Using Wolbachia to control rice planthopper populations: progress and challenges.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1244239}, pmid = {37779725}, issn = {1664-302X}, abstract = {Wolbachia have been developed as a tool for protecting humans from mosquito populations and mosquito-borne diseases. The success of using Wolbachia relies on the facts that Wolbachia are maternally transmitted and that Wolbachia-induced cytoplasmic incompatibility provides a selective advantage to infected over uninfected females, ensuring that Wolbachia rapidly spread through the target pest population. Most transinfected Wolbachia exhibit a strong antiviral response in novel hosts, thus making it an extremely efficient technique. Although Wolbachia has only been used to control mosquitoes so far, great progress has been made in developing Wolbachia-based approaches to protect plants from rice pests and their associated diseases. Here, we synthesize the current knowledge about the important phenotypic effects of Wolbachia used to control mosquito populations and the literature on the interactions between Wolbachia and rice pest planthoppers. Our aim is to link findings from Wolbachia-mediated mosquito control programs to possible applications in planthoppers.}, } @article {pmid37768955, year = {2023}, author = {Chebbah, D and Hamarsheh, O and Sereno, D and Elissa, N and Brun, S and Jan, J and Izri, A and Akhoundi, M}, title = {Molecular characterization and genetic diversity of Wolbachia endosymbionts in bed bugs (Hemiptera; Cimicidae) collected in Paris.}, journal = {PloS one}, volume = {18}, number = {9}, pages = {e0292229}, pmid = {37768955}, issn = {1932-6203}, mesh = {Animals ; *Bedbugs/genetics ; *Wolbachia/genetics ; Phylogeny ; Polymerase Chain Reaction ; Nymph ; Genetic Variation ; }, abstract = {PURPOSE: This study aimed to investigate the genetic diversity of Wolbachia in field-caught bed bug species in Paris areas.

METHODS: The bed bug specimens were captured from various infested localities in Paris and surrounding cities. They belonged to diverse life stages, including egg, nymph, and adult. They were then identified using morphological and molecular approaches. Furthermore, Wolbachia was detected, and its genetic diversity was investigated by conventional PCR of 16S-rRNA and Wolbachia surface protein (wsp) genes.

RESULTS: A total of 256 bed bug specimens belonging to various life stages [adult (183 specimens), nymph (48), and egg (25)] were captured from seven private apartments, five social apartments, three houses, two immigrant residences, and one retirement home situated in 10 districts of Paris and 8 surrounding cities. They were identified as Cimex lectularius (237 specimens) and C. hemipterus (19) using morphological and molecular approaches. The presence and diversity of Wolbachia were ascertained by targeting 16S-rRNA and wsp genes. Based on molecular analysis, 182 and 148 out of 256 processed specimens were positive by amplifying 16S-rRNA and wsp fragments, respectively. The inferred phylogenetic analysis with 16S-rRNA and wsp sequences displayed monophyletic Wolbachia strains clustering each one in three populations. The median-joining network, including the Wolbachia 16S-rRNA and wsp sequences of C. lectularius and C. hemipterous specimens, indicated a significant genetic differentiation among these populations in Paris areas which was consent with Neighbor-Joining analyses. A phylogenetic analysis of our heterogenic Wolbachia sequences with those reported from other arthropod species confirmed their belonging to supergroup F. Moreover, no difference between Wolbachia sequences from eggs, nymphs, and adults belonging to the same clade and between Wolbachia sequences of C. lectularius and C. hemipterus were observed after sequence alignment. Furthermore, no significant correlation was found between multiple geographical locations (or accomodation type) where bed bugs were collected and the genetic diversity of Wolbachia.

CONCLUSIONS: We highlight a significant heterogeneity within Wolbachia symbionts detected in C. lectularius and C. hemipterus. No correlation between Wolbachia species and bed bug species (C. lectularius versus C. hemipterus), physiological stages (egg, nymph, and adult), and sampling location was recorded in this study.}, } @article {pmid37744653, year = {2023}, author = {Thi Hue Kien, D and Edenborough, K and da Silva Goncalves, D and Thuy Vi, T and Casagrande, E and Thi Le Duyen, H and Thi Long, V and Thi Dui, L and Thi Tuyet Nhu, V and Thi Giang, N and Thi Xuan Trang, H and Lee, E and Donovan-Banfield, I and Thi Thuy Van, H and Minh Nguyet, N and Thanh Phong, N and Van Vinh Chau, N and Wills, B and Yacoub, S and Flores, H and Simmons, C}, title = {Genome evolution of dengue virus serotype 1 under selection by Wolbachia pipientis in Aedes aegypti mosquitoes.}, journal = {Virus evolution}, volume = {9}, number = {1}, pages = {vead016}, pmid = {37744653}, issn = {2057-1577}, abstract = {The introgression of antiviral strains of Wolbachia into Aedes aegypti mosquito populations is a public health intervention for the control of dengue. Plausibly, dengue virus (DENV) could evolve to bypass the antiviral effects of Wolbachia and undermine this approach. Here, we established a serial-passage system to investigate the evolution of DENV in Ae. aegypti mosquitoes infected with the wMel strain of Wolbachia. Using this system, we report on virus genetic outcomes after twenty passages of serotype 1 of DENV (DENV-1). An amino acid substitution, E203K, in the DENV-1 envelope protein was more frequently detected in the consensus sequence of virus populations passaged in wMel-infected Ae. aegypti than wild-type counterparts. Positive selection at residue 203 was reproducible; it occurred in passaged virus populations from independent DENV-1-infected patients and also in a second, independent experimental system. In wild-type mosquitoes and human cells, the 203K variant was rapidly replaced by the progenitor sequence. These findings provide proof of concept that wMel-associated selection of virus populations can occur in experimental conditions. Field-based studies are needed to explore whether wMel imparts selective pressure on DENV evolution in locations where wMel is established.}, } @article {pmid37738428, year = {2024}, author = {Thia, JA and Endersby-Harshman, N and Collier, S and Nassar, MS and Tawfik, EA and Alfageeh, MB and Elfekih, S and Hoffmann, AA}, title = {Mitochondrial DNA variation in Aedes aegypti (Diptera: Culicidae) mosquitoes from Jeddah, Saudi Arabia.}, journal = {Journal of medical entomology}, volume = {61}, number = {1}, pages = {250-256}, pmid = {37738428}, issn = {1938-2928}, mesh = {Animals ; *Aedes ; Saudi Arabia ; Prospective Studies ; Mutation ; DNA, Mitochondrial ; *Wolbachia/genetics ; Mosquito Vectors/genetics ; }, abstract = {Wolbachia (Hertig 1936) (Rickettsiales: Ehrlichiaceae) has emerged as a valuable biocontrol tool in the fight against dengue by suppressing the transmission of the virus through mosquitoes. Monitoring the dynamics of Wolbachia is crucial for evaluating the effectiveness of release programs. Mitochondrial (mtDNA) markers serve as important tools for molecular tracking of infected mitochondrial backgrounds over time but require an understanding of the variation in release sites. In this study, we investigated the mitochondrial lineages of Aedes aegypti (Linnaeus 1762) in Jeddah, Saudi Arabia, which is a prospective release site for the "wAlbBQ" Wolbachia-infected strain of this mosquito species. We employed a combination of comprehensive mitogenomic analysis (including all protein-coding genes) and mtDNA marker analysis (cox1 and nad5) using data collected from Jeddah. We combined our mitogenome and mtDNA marker data with those from previous studies to place mitochondrial variation in Saudi Arabia into a broader global context. Our findings revealed the presence of 4 subclades that can be broadly categorized into 2 major mitochondrial lineages. Ae. aegypti mosquitoes from Jeddah belonged to both major lineages. Whilst mitogenomic data offered a higher resolution for distinguishing Jeddah mosquitoes from the wAlbBQ strain, the combination of cox1 and nad5 mtDNA markers alone proved to be sufficient. This study provides the first important characterization of Ae. aegypti mitochondrial lineages in Saudi Arabia and offers essential baseline information for planning future molecular monitoring efforts during the release of Wolbachia-infected mosquitoes.}, } @article {pmid37729233, year = {2023}, author = {Soto, A and De Coninck, L and Devlies, AS and Van De Wiele, C and Rosales Rosas, AL and Wang, L and Matthijnssens, J and Delang, L}, title = {Belgian Culex pipiens pipiens are competent vectors for West Nile virus while Culex modestus are competent vectors for Usutu virus.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {9}, pages = {e0011649}, pmid = {37729233}, issn = {1935-2735}, mesh = {Animals ; Female ; Humans ; *West Nile virus/genetics ; *Culex ; Belgium ; *Flavivirus/genetics ; *Culicidae ; Mosquito Vectors ; *Arboviruses ; *West Nile Fever ; }, abstract = {BACKGROUND: West Nile virus (WNV) and Usutu virus (USUV) are emerging arthropod-borne viruses (arboviruses) in Europe transmitted by Culex mosquitoes. In Belgium, it is currently unknown which Culex species are competent vectors for WNV or USUV and if these mosquitoes carry Wolbachia, an endosymbiotic bacterium that can block arbovirus transmission. The aims of our study were to measure the vector competence of Belgian Culex mosquitoes to WNV and USUV and determine if a naturally acquired Wolbachia infection can influence virus transmission.

Female Culex mosquitoes were captured from urban and peri-urban sites in Leuven, Belgium and offered an infectious bloodmeal containing WNV lineage 2, USUV European (EU) lineage 3, or USUV African (AF) lineage 3. Blood-fed females were incubated for 14 days at 25°C after which the body, head, and saliva were collected to measure infection, dissemination, and transmission rates as well as transmission efficiency. Mosquito species were identified by qRT-PCR or Sanger sequencing, the presence of infectious virus in mosquitoes was confirmed by plaque assays, and viral genome copies were quantified by qRT-PCR. Culex pipiens pipiens were able to transmit WNV (4.3% transmission efficiency, n = 2/47) but not USUV (EU lineage: n = 0/56; AF lineage: n = 0/37). In contrast, Culex modestus were able to transmit USUV (AF lineage: 20% transmission efficiency, n = 1/5) but not WNV (n = 0/6). We found that the presence or absence of Wolbachia was species-dependent and did not associate with virus transmission.

CONCLUSIONS/SIGNIFICANCE: This is the first report that Belgian Culex mosquitoes can transmit both WNV and USUV, forewarning the risk of human transmission. More research is needed to understand the potential influence of Wolbachia on arbovirus transmission in Culex modestus mosquitoes.}, } @article {pmid37720159, year = {2023}, author = {Giordano, R and Weber, EP and Mitacek, R and Flores, A and Ledesma, A and De, AK and Herman, TK and Soto-Adames, FN and Nguyen, MQ and Hill, CB and Hartman, GL}, title = {Patterns of asexual reproduction of the soybean aphid, Aphis glycines (Matsumura), with and without the secondary symbionts Wolbachia and Arsenophonus, on susceptible and resistant soybean genotypes.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1209595}, pmid = {37720159}, issn = {1664-302X}, abstract = {Plant breeding is used to develop crops with host resistance to aphids, however, virulent biotypes often develop that overcome host resistance genes. We tested whether the symbionts, Arsenophonus (A) and Wolbachia (W), affect virulence and fecundity in soybean aphid biotypes Bt1 and Bt3 cultured on whole plants and detached leaves of three resistant, Rag1, Rag2 and Rag1 + 2, and one susceptible, W82, soybean genotypes. Whole plants and individual aphid experiments of A. glycines with and without Arsenophonus and Wolbachia did not show differences in overall fecundity. Differences were observed in peak fecundity, first day of deposition, and day of maximum nymph deposition of individual aphids on detached leaves. Bt3 had higher fecundity than Bt1 on detached leaves of all plant genotypes regardless of bacterial profile. Symbionts did not affect peak fecundity of Bt1 but increased it in Bt3 (A+W+) and all Bt3 strains began to deposit nymphs earlier than the Bt1 (A+W-). Arsenophonus in Bt1 delayed the first day of nymph deposition in comparison to aposymbiotic Bt1 except when reared on Rag1 + 2. For the Bt1 and Bt3 strains, symbionts did not result in a significant difference in the day they deposited the maximum number of nymphs nor was there a difference in survival or variability in number of nymphs deposited. Variability of number of aphids deposited was higher in aphids feeding on resistant plant genotypes. The impact of Arsenophonus on soybean aphid patterns of fecundity was dependent on the aphid biotype and plant genotype. Wolbachia alone had no detectable impact but may have contributed to the increased fecundity of Bt3 (A+W+). An individual based model, using data from the detached leaves experiment and with intraspecific competition removed, found patterns similar to those observed in the greenhouse and growth chamber experiments including a significant interaction between soybean genotype and aphid strain. Combining individual data with the individual based model of population growth isolated the impact of fecundity and host resistance from intraspecific competition and host health. Changes to patterns of fecundity, influenced by the composition and concentration of symbionts, may contribute to competitive interactions among aphid genotypes and influence selection on virulent aphid populations.}, } @article {pmid37716961, year = {2023}, author = {Mfopit, YM and Engel, JS and Chechet, GD and Ibrahim, MAM and Signaboubo, D and Achukwi, DM and Mamman, M and Balogun, EO and Shuaibu, MN and Kabir, J and Kelm, S}, title = {Molecular detection of Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts in wild population of tsetse flies collected in Cameroon, Chad and Nigeria.}, journal = {BMC microbiology}, volume = {23}, number = {1}, pages = {260}, pmid = {37716961}, issn = {1471-2180}, mesh = {Animals ; *Tsetse Flies ; *Wolbachia/genetics ; Cameroon ; Chad ; Nigeria ; *Glossinidae ; *Trypanosomiasis, African ; *Spiroplasma/genetics ; }, abstract = {BACKGROUND: Tsetse flies are cyclical vectors of African trypanosomiasis (AT). The flies have established symbiotic associations with different bacteria that influence certain aspects of their physiology. Vector competence of tsetse flies for different trypanosome species is highly variable and is suggested to be affected by bacterial endosymbionts amongst other factors. Symbiotic interactions may provide an avenue for AT control. The current study provided prevalence of three tsetse symbionts in Glossina species from Cameroon, Chad and Nigeria.

RESULTS: Tsetse flies were collected and dissected from five different locations. DNA was extracted and polymerase chain reaction used to detect presence of Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts, using species specific primers. A total of 848 tsetse samples were analysed: Glossina morsitans submorsitans (47.52%), Glossina palpalis palpalis (37.26%), Glossina fuscipes fuscipes (9.08%) and Glossina tachinoides (6.13%). Only 95 (11.20%) were infected with at least one of the three symbionts. Among infected flies, six (6.31%) had Wolbachia and Spiroplasma mixed infection. The overall symbiont prevalence was 0.88, 3.66 and 11.00% respectively, for Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts. Prevalence varied between countries and tsetse fly species. Neither Spiroplasma species nor S. glossinidius were detected in samples from Cameroon and Nigeria respectively.

CONCLUSION: The present study revealed, for the first time, presence of Spiroplasma species infections in tsetse fly populations in Chad and Nigeria. These findings provide useful information on repertoire of bacterial flora of tsetse flies and incite more investigations to understand their implication in the vector competence of tsetse flies.}, } @article {pmid37715236, year = {2023}, author = {ElKraly, OA and Awad, M and El-Saadany, HM and Hassanein, SE and Elrahman, TA and Elnagdy, SM}, title = {Impact of gut microbiota composition on black cutworm, Agrotis ipsilon (hufnagel) metabolic indices and pesticide degradation.}, journal = {Animal microbiome}, volume = {5}, number = {1}, pages = {44}, pmid = {37715236}, issn = {2524-4671}, abstract = {Endosymbionts are known to have significant effects on their insect hosts, including nutrition, reproduction, and immunity. Insects gut microbiota is a critical component that affects their physiological and behavioral characteristics. The black cutworm (BCW), Agrotis ipsilon, is an economically important lepidopteran pest that has a diverse gut microbiome composed of nine species belonging to three phyla: Proteobacteria, Actinobacteria, and Firmicutes. This study was conducted to investigate the diversity of gut bacteria isolated from BCW larvae and moths and their effects on metabolism and pesticide degradation. The bacterial isolates were identified using the 16 S rRNA gene. The study showed that the gut microbiome composition significantly affected the metabolism of BCW larvae. Based on the screening results of synthesis of digestive enzymes and pesticide degradation, Brachybacterium conglomeratum and Glutamicibacter sp were selected to perform the remaining experiments as single isolates and consortium. The consortium-fed larvae showed high metabolic indices compared to antibiotic-fed larvae and the control. The gut bacteria were also shown to degrade three pesticide groups. Concerns regarding the health risk of chlorpyrifos have been raised due to its extensive use in agriculture. The isolated B. conglomeratum was more effective in chlorpyrifos degradation than the consortium. Furthermore, the study also examined the presence of sex related endosymbionts (Wolbachia, Spiroplasma, and Rickettsia) in the reproductive tissues of adults. The outcomes demonstrated that none of the examined endosymbionts existed. In conclusion, the study highlights the importance of the gut microbiome in insect physiology and behavior and its potential applications in biotechnology. It provides insights into developing eco-friendly pest control and bioremediation strategies using gut bacteria.}, } @article {pmid37706052, year = {2023}, author = {Vinayagam, S and Nirmolia, T and Chetry, S and Kumar, NP and Saini, P and Bhattacharyya, DR and Bhowmick, IP and Sattu, K and Patgiri, SJ}, title = {Molecular Evidence of Wolbachia Species in Wild-Caught Aedes albopictus and Aedes aegypti Mosquitoes in Four States of Northeast India.}, journal = {Journal of tropical medicine}, volume = {2023}, number = {}, pages = {6678627}, pmid = {37706052}, issn = {1687-9686}, abstract = {Wolbachia, a Gram-negative intracellular bacterium, naturally infects many arthropods, including mosquito vectors responsible for the spread of arboviral diseases such as Zika, chikungunya, and dengue fever. Certain Wolbachia strains are involved in inhibiting arbovirus replication in mosquitoes, and this phenomenon is currently being studied to combat disease vectors. A study was conducted in four states in north-eastern India to investigate the presence of natural Wolbachia infection in wild-caught Aedes albopictus and Aedes aegypti mosquitoes, the established vectors of dengue. The detection of a Wolbachia infection was confirmed by nested PCR and sequencing in the two mosquito species Ae. aegypti and Ae. albopictus. Positivity rates observed in Ae. aegypti and Ae. albopictus pools were 38% (44 of 115) and 85% (41 of 48), respectively, and the difference was significant (chi-square = 28.3174, p = 0.00000010). Sequencing revealed that all detected Wolbachia strains belonged to supergroup B. Although Wolbachia infection in Ae. aegypti has been previously reported from India, no such reports are available from north-eastern India. Data on naturally occurring Wolbachia strains are essential for selecting the optimal strain for the development of Wolbachia-based control measures. This information will be helpful for the future application of Wolbachia-based vector control measures in this part of the country.}, } @article {pmid37696983, year = {2023}, author = {Ogunlade, ST and Adekunle, AI and Meehan, MT and McBryde, ES}, title = {Quantifying the impact of Wolbachia releases on dengue infection in Townsville, Australia.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {14932}, pmid = {37696983}, issn = {2045-2322}, mesh = {Animals ; *Wolbachia ; Mosquito Vectors ; Australia/epidemiology ; Queensland/epidemiology ; *Culicidae ; *Dengue/epidemiology/prevention & control ; }, abstract = {From October 2014 to February 2019, local authorities in Townsville, North Queensland, Australia continually introduced Wolbachia-infected mosquitoes to control seasonal outbreaks of dengue infection. In this study, we develop a mathematical modelling framework to estimate the effectiveness of this intervention as well as the relative dengue transmission rates of Wolbachia-infected and wild-type mosquitoes. We find that the transmission rate of Wolbachia-infected mosquitoes is reduced approximately by a factor of 20 relative to the uninfected wild-type population. In addition, the Townsville Wolbachia release program led to a 65% reduction in predicted dengue incidence during the release period and over 95% reduction in the 24 months that followed. Finally, to investigate the potential impact of other Wolbachia release programs, we use our estimates of relative transmissibility to calculate the relationship between the reproductive number of dengue and the proportion of Wolbachia-infected mosquitoes in the vector population.}, } @article {pmid37695720, year = {2023}, author = {Kryukova, NA and Kryukov, VY and Polenogova, OV and Chertkova, ЕА and Tyurin, MV and Rotskaya, UN and Alikina, T and Kabilov, МR and Glupov, VV}, title = {The endosymbiotic bacterium Wolbachia (Rickettsiales) alters larval metabolism of the parasitoid Habrobracon hebetor (Hymenoptera: Braconidae).}, journal = {Archives of insect biochemistry and physiology}, volume = {114}, number = {4}, pages = {e22053}, doi = {10.1002/arch.22053}, pmid = {37695720}, issn = {1520-6327}, support = {//Federal Fundamental Scientific Research Program/ ; //Russian Science Foundation/ ; 23-24-00259//The Russian Science Foundation/ ; 1021051703454-5-1.6.12//The Federal Fundamental Scientific Research Program/ ; }, mesh = {Animals ; Larva/metabolism ; *Hymenoptera ; *Wasps/metabolism ; *Wolbachia ; Rickettsiales ; *Moths/metabolism ; }, abstract = {Infection of intestinal tissues with Wolbachia has been found in Habrobracon hebetor. There are not many studies on the relationship between Habrobracon and Wolbachia, and they focus predominantly on the sex index of an infected parasitoid, its fertility, and behavior. The actual role of Wolbachia in the biology of Habrobracon is not yet clear. The method of complete eradication of Wolbachia in the parasitoid was developed here, and effects of the endosymbiont on the host's digestive metabolism were compared between two lines of the parasitoid (Wolbachia-positive and Wolbachia-negative). In the gut of Wolbachia[+] larvae, lipases' activity was higher almost twofold, and activities of acid proteases, esterases, and trehalase were 1.5-fold greater than those in the Wolbachia[-] line. Analyses of larval homogenates revealed that Wolbachia[+] larvae accumulate significantly more lipids and have a lower amount of pyruvate as compared to Wolbachia[-] larvae. The presented results indicate significant effects of the intracellular symbiotic bacterium Wolbachia on the metabolism of H. hebetor larvae and on the activity of its digestive enzymes.}, } @article {pmid37693004, year = {2023}, author = {Ning, SF and Huo, LX and Lv, L and Wang, Y and Zhang, LS and Che, WN and Dong, H and Zhou, JC}, title = {The identification and expression pattern of the sex determination genes and their sex-specific variants in the egg parasitoid Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae).}, journal = {Frontiers in physiology}, volume = {14}, number = {}, pages = {1243753}, pmid = {37693004}, issn = {1664-042X}, abstract = {Introduction: Trichogramma wasps are egg parasitoids of agricultural lepidopteran pests. The sex of Trichogramma is determined by its ploidy as well as certain sex ratio distorters, such as the endosymbiotic bacteria Wolbachia spp. and the paternal sex ratio (PSR) chromosome. The sex determination systems of hymenopterans, such as Trichogramma spp., involve cascades of the genes transformer (tra), transformer-2 (tra2), and doublesex (dsx) and are associated with sex-specific tra and dsx splicing. First, these genes and their sex-specific variants must be identified to elucidate the interactions between the sex ratio disorders and the sex determination mechanism of Trichogramma. Methods: Here, we characterized the sex determination genes tra, tra2, and dsx in Trichogramma dendrolimi. Sex-specific tra and dsx variants were detected in cDNA samples obtained from both male and female Trichogramma wasps. They were observed in the early embryos (1-10 h), late embryos (12-20 h), larvae (32 h and 48 h), pre-pupae (96 h), and pupae (144 h, 168 h, 192 h, and 216 h) of both male and female T. dendrolimi offspring. Results: We detected female-specific tra variants throughout the entire early female offspring stage. The male-specific variant began to express at 9-10 h as the egg was not fertilized. However, we did not find any maternally derived, female-specific tra variant in the early male embryo. This observation suggests that the female-specific tra variant expressed in the female embryo at 1-9 h may not have originated from the maternal female wasp. Discussion: The present study might be the first to identify the sex determination genes and sex-specific gene splicing in Trichogramma wasps. The findings of this study lay the foundation for investigating the sex determination mechanisms of Trichogramma and other wasps. They also facilitate sex identification in immature T. dendrolimi and the application of this important egg parasitoid in biological insect pest control programs.}, } @article {pmid37686049, year = {2023}, author = {Zhang, Z and Zhang, J and Chen, Q and He, J and Li, X and Wang, Y and Lu, Y}, title = {Complete De Novo Assembly of Wolbachia Endosymbiont of Frankliniella intonsa.}, journal = {International journal of molecular sciences}, volume = {24}, number = {17}, pages = {}, pmid = {37686049}, issn = {1422-0067}, support = {31672031, 32272537//National Natural Science Foundation of China/ ; 2021C02003//Key Research and Development Program of Zhejiang Province, China/ ; 2022YFD1401204, 2022YFC2601405//Key R&D Program of China/ ; }, mesh = {Animals ; *Thysanoptera ; *Wolbachia/genetics ; Flowers ; *Nanopores ; Prophages ; RNA, Ribosomal ; }, abstract = {As an endosymbiont, Wolbachia exerts significant effects on the host, including on reproduction, immunity, and metabolism. However, the study of Wolbachia in Thysanopteran insects, such as flower thrips Frankliniella intonsa, remains limited. Here, we assembled a gap-free looped genome assembly of Wolbachia strain wFI in a length of 1,463,884 bp (GC content 33.80%), using Nanopore long reads and Illumina short reads. The annotation of wFI identified a total of 1838 protein-coding genes (including 85 pseudogenes), 3 ribosomal RNAs (rRNAs), 35 transfer RNAs (tRNAs), and 1 transfer-messenger RNA (tmRNA). Beyond this basic description, we identified mobile genetic elements, such as prophage and insertion sequences (ISs), which make up 17% of the entire wFI genome, as well as genes involved in riboflavin and biotin synthesis and metabolism. This research lays the foundation for understanding the nutritional mutualism between Wolbachia and flower thrips. It also serves as a valuable resource for future studies delving into the intricate interactions between Wolbachia and its host.}, } @article {pmid37680477, year = {2023}, author = {Yeo, H and Tan, HZ and Tang, Q and Tan, TRH and Puniamoorthy, N and Rheindt, FE}, title = {Dense residential areas promote gene flow in dengue vector mosquito Aedes albopictus.}, journal = {iScience}, volume = {26}, number = {9}, pages = {107577}, pmid = {37680477}, issn = {2589-0042}, abstract = {Aedes albopictus is a successful disease vector due to its ability to survive in a wide range of habitats. Despite its ubiquity and impact on public health, little is known about its differential gene flow capabilities across different city habitats. We obtained a comprehensive dataset of >27,000 genome-wide DNA markers across 105 wild-caught Ae. albopictus individuals from Singapore, a dengue-endemic tropical city with heterogeneous landscapes from densely populated urban areas to forests. Despite Singapore's challenging small-scale heterogeneity, our landscape-genomic approach indicated that dense urban areas are characterized by higher Aedes gene flow rates than managed parks and forests. We documented the incidence of Wolbachia infections of Ae. albopictus involving two strains (wAlbA and wAlbB). Our results dispel the misconception that substantial dispersal of Ae. albopictus is limited to urban greenery, with wide implications for vector management and critical insights into urban planning strategies to combat dengue transmission.}, } @article {pmid37669272, year = {2023}, author = {Lau, MJ and Dutra, HLC and Jones, MJ and McNulty, BP and Diaz, AM and Ware-Gilmore, F and McGraw, EA}, title = {Jamestown Canyon virus is transmissible by Aedes aegypti and is only moderately blocked by Wolbachia co-infection.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {9}, pages = {e0011616}, pmid = {37669272}, issn = {1935-2735}, support = {R01 AI143758/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Humans ; *Aedes ; *Encephalitis Virus, California ; *Deer ; *Wolbachia ; *Coinfection ; Mosquito Vectors ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {Jamestown Canyon virus (JCV), a negative-sense arbovirus, is increasingly common in the upper Midwest of the USA. Transmitted by a range of mosquito genera, JCV's primary amplifying host is white-tailed deer. Aedes aegypti is responsible for transmitting various positive-sense viruses globally including dengue (DENV), Zika, chikungunya, and Yellow Fever. Ae. aegypti's distribution, once confined to the tropics, is expanding, in part due to climate change. Wolbachia, an insect endosymbiont, limits the replication of co-infecting viruses inside insects. The release and spread of the symbiont into Ae. aegypti populations have been effective in reducing transmission of DENV to humans, although the mechanism of Wolbachia-mediated viral blocking is still poorly understood. Here we explored JCV infection potential in Ae. aegypti, the nature of the vector's immune response, and interactions with Wolbachia infection. We show that Ae. aegypti is highly competent for JCV, which grows to high loads and rapidly reaches the saliva after an infectious blood meal. The mosquito immune system responds with strong induction of RNAi and JAK/STAT. Neither the direct effect of viral infection nor the energetic investment in immunity appears to affect mosquito longevity. Wolbachia infection blocked JCV only in the early stages of infection. Wolbachia-induced immunity was small compared to that of JCV, suggesting innate immune priming does not likely explain blocking. We propose two models to explain why Wolbachia's blocking of negative-sense viruses like JCV may be less than that of positive-sense viruses, relating to the slowdown of host protein synthesis and the triggering of interferon-like factors like Vago. In conclusion, we highlight the risk for increased human disease with the predicted future overlap of Ae. aegypti and JCV ranges. We suggest that with moderate Wolbachia-mediated blocking and distinct biology, negative-sense viruses represent a fruitful comparator model to other viruses for understanding blocking mechanisms in mosquitoes.}, } @article {pmid37658881, year = {2023}, author = {Owashi, Y and Minami, T and Kikuchi, T and Yoshida, A and Nakano, R and Kageyama, D and Adachi-Hagimori, T}, title = {Microbiome of Zoophytophagous Biological Control Agent Nesidiocoris tenuis.}, journal = {Microbial ecology}, volume = {86}, number = {4}, pages = {2923-2933}, pmid = {37658881}, issn = {1432-184X}, mesh = {Humans ; Animals ; Biological Control Agents ; *Hemiptera/genetics ; RNA, Ribosomal, 16S/genetics ; *Rickettsia/genetics ; *Spiroplasma ; *Wolbachia/genetics ; *Microbiota ; Symbiosis ; }, abstract = {Many insects are associated with endosymbionts that influence the feeding, reproduction, and distribution of their hosts. Although the small green mirid, Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae), a zoophytophagous predator that feeds on plants as well as arthropods, is a globally important biological control agent, its microbiome has not been sufficiently studied. In the present study, we assessed the microbiome variation in 96 N. tenuis individuals from 14 locations throughout Japan, based on amplicon sequencing of the 16S ribosomal RNA gene. Nine major bacteria associated with N. tenuis were identified: Rickettsia, two strains of Wolbachia, Spiroplasma, Providencia, Serratia, Pseudochrobactrum, Lactococcus, and Stenotrophomonas. Additionally, a diagnostic PCR analysis for three typical insect reproductive manipulators, Rickettsia, Wolbachia, and Spiroplasma, was performed on a larger sample size (n = 360) of N. tenuis individuals; the most prevalent symbiont was Rickettsia (69.7%), followed by Wolbachia (39.2%) and Spiroplasma (6.1%). Although some symbionts were co-infected, their prevalence did not exhibit any specific tendency, such as a high frequency in specific infection combinations. The infection frequency of Rickettsia was significantly correlated with latitude and temperature, while that of Wolbachia and Spiroplasma was significantly correlated with host plants. The predominance of these bacteria and the absence of obligate symbionts suggested that the N. tenuis microbiome is typical for predatory arthropods rather than sap-feeding insects. Rickettsia and Wolbachia were vertically transmitted rather than horizontally transmitted from the prey. The functional validation of each symbiont would be warranted to develop N. tenuis as a biological control agent.}, } @article {pmid37653429, year = {2023}, author = {Duong Thi Hue, K and da Silva Goncalves, D and Tran Thuy, V and Thi Vo, L and Le Thi, D and Vu Tuyet, N and Nguyen Thi, G and Huynh Thi Xuan, T and Nguyen Minh, N and Nguyen Thanh, P and Yacoub, S and Simmons, CP}, title = {Wolbachia wMel strain-mediated effects on dengue virus vertical transmission from Aedes aegypti to their offspring.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {308}, pmid = {37653429}, issn = {1756-3305}, mesh = {Female ; Animals ; *Aedes ; *Dengue Virus ; *Wolbachia ; Infectious Disease Transmission, Vertical ; Laboratories ; }, abstract = {BACKGROUND: Dengue virus serotypes (DENV-1 to -4) can be transmitted vertically in Aedes aegpti mosquitoes. Whether infection with the wMel strain of the endosymbiont Wolbachia can reduce the incidence of vertical transmission of DENV from infected females to their offspring is not well understood.

METHODS: A laboratory colony of Vietnamese Ae. aegypti, both with and without wMel infection, were infected with DENV-1 by intrathoracic injection (IT) to estimate the rate of vertical transmission (VT) of the virus. VT in the DENV-infected mosquitoes was calculated via the infection rate estimation from mosquito pool data using maximum likelihood estimation (MLE).

RESULTS: In 6047 F1 Vietnamese wild-type Ae. aegypti, the MLE of DENV-1 infection was 1.49 per 1000 mosquitoes (95% confidence interval [CI] 0.73-2.74). In 5500 wMel-infected Ae. aegypti, the MLE infection rate was 0 (95% CI 0-0.69). The VT rates between mosquito lines showed a statistically significant difference.

CONCLUSIONS: The results reinforce the view that VT is a rare event in wild-type mosquitoes and that infection with wMel is effective in reducing VT.}, } @article {pmid37652566, year = {2023}, author = {Cavany, S and Huber, JH and Wieler, A and Tran, QM and Alkuzweny, M and Elliott, M and España, G and Moore, SM and Perkins, TA}, title = {Does ignoring transmission dynamics lead to underestimation of the impact of interventions against mosquito-borne disease?.}, journal = {BMJ global health}, volume = {8}, number = {8}, pages = {}, pmid = {37652566}, issn = {2059-7908}, support = {R35 GM143029/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Humans ; *Vector Borne Diseases/prevention & control/transmission ; Culicidae ; Bias ; Models, Biological ; }, abstract = {New vector-control technologies to fight mosquito-borne diseases are urgently needed, the adoption of which depends on efficacy estimates from large-scale cluster-randomised trials (CRTs). The release of Wolbachia-infected mosquitoes is one promising strategy to curb dengue virus (DENV) transmission, and a recent CRT reported impressive reductions in dengue incidence following the release of these mosquitoes. Such trials can be affected by multiple sources of bias, however. We used mathematical models of DENV transmission during a CRT of Wolbachia-infected mosquitoes to explore three such biases: human movement, mosquito movement and coupled transmission dynamics between trial arms. We show that failure to account for each of these biases would lead to underestimated efficacy, and that the majority of this underestimation is due to a heretofore unrecognised bias caused by transmission coupling. Taken together, our findings suggest that Wolbachia-infected mosquitoes could be even more promising than the recent CRT suggested. By emphasising the importance of accounting for transmission coupling between arms, which requires a mathematical model, we highlight the key role that models can play in interpreting and extrapolating the results from trials of vector control interventions.}, } @article {pmid37649415, year = {2023}, author = {Martínez-Burgos, M and Lozano-Sardaneta, YN and Rodríguez-Rojas, JJ and Gómez-Rivera, ÁS and Canto-Mis, KL and Flores-Escobar, E and Mis-Ávila, PC and Correa-Morales, F and Becker, I}, title = {Species diversity and detection of pathogens in phlebotomine sand flies collected from forest management areas of Quintana Roo, Mexico.}, journal = {Medical and veterinary entomology}, volume = {37}, number = {4}, pages = {845-858}, doi = {10.1111/mve.12691}, pmid = {37649415}, issn = {1365-2915}, mesh = {Animals ; *Psychodidae ; Mexico ; Insect Vectors ; *Phlebotomus ; Forests ; }, abstract = {Sand flies have expanded their areas of distribution, thereby increasing the risk of pathogen transmission in non-endemic areas. To establish efficient prevention and control strategies for the transmission of vector-borne pathogens, it is important to understand seasonal dynamics of their vectors. In Mexico, there are several areas where the contact between sand flies, hosts and reservoirs favours the transmission of the pathogen. We compared sand fly communities in a forest management area and a conserved area in Noh-Bec, Quintana Roo, Mexico. The analysis included species diversity, activity peaks and molecular detection of pathogens. Sand flies were collected from November to December 2021 and April to May 2022, during 84 night-traps. The conserved area showed higher numbers and greater species heterogeneity of sand flies as compared with the other sites. The β-diversity analysis revealed that sites disturbed by logging (S1, S2, S3) had greater similarity (90%) in their sand fly species composition than a conserved area (S4) (similarity = 36%). Although none of the specimens were infected with Leishmania, we detected Wolbachia (19.4%) in all four sites, as well as Bartonella (3.25%) only in the disturbed sites. Further studies on the dynamics of sand fly populations and their association with pathogens are necessary.}, } @article {pmid37647311, year = {2023}, author = {Wei, TL and Zheng, YP and Wang, ZH and Shang, YX and Pei, MS and Liu, HN and Yu, YH and Shi, QF and Jiang, DM and Guo, DL}, title = {Comparative microbiome analysis reveals the variation in microbial communities between 'Kyoho' grape and its bud mutant variety.}, journal = {PloS one}, volume = {18}, number = {8}, pages = {e0290853}, pmid = {37647311}, issn = {1932-6203}, mesh = {*Chlorobi ; *Vitis ; *Microbiota/genetics ; *Anaplasmataceae ; Enterobacteriaceae ; }, abstract = {Microbes are an important part of the vineyard ecosystem, which significantly influence the quality of grapes. Previously, we identified a bud mutant variety (named 'Fengzao') from 'Kyoho' grapes. The variation of microbial communities in grape and its bud mutant variety has not been studied yet. So, in this study, with the samples of both 'Fengzao' and 'Kyoho', we conducted high-throughput microbiome sequencing and investigated their microbial communities in different tissues. Obvious differences were observed in the microbial communities between 'Fengzao' and 'Kyoho'. The fruit and the stem are the tissues with relatively higher abundance of microbes, while the leaves contained less microbes. The fruit and the stem of 'Kyoho' and the stem of 'Fengzao' had relatively higher species diversity based on the alpha diversity analysis. Proteobacteria, Enterobacteriaceae and Rhodobacteraceae had significantly high abundance in 'Fengzao'. Firmicutes and Pseudomonas were highly abundant in the stems of 'Kyoho', and family of Spirochaetaceae, Anaplasmataceae, Chlorobiaceae, and Sphingomonadaceae, and genera of Spirochaeta, Sphingomonas, Chlorobaculum and Wolbachia were abundant in the fruits of 'Kyoho'. These identified microbes are main components of the microbial communities, and could be important regulators of grapevine growth and development. This study revealed the differences in the microbial compositions between 'Kyoho' and its bud mutant, and these identified microbes will be significant resources for the future researches on the quality regulation and disease control of grapevines.}, } @article {pmid37645949, year = {2023}, author = {Wenzel, M and Aquadro, CF}, title = {Wolbachia infection at least partially rescues the fertility and ovary defects of several new Drosophila melanogaster bag of marbles protein-coding mutants.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2023.03.20.532813}, pmid = {37645949}, abstract = {UNLABELLED: The D. melanogaster protein coding gene bag of marbles (bam) plays a key role in early male and female reproduction by forming complexes with partner proteins to promote differentiation in gametogenesis. Like another germline gene, Sex lethal , bam genetically interacts with the endosymbiont Wolbachia , as Wolbachia rescues the reduced fertility of a bam hypomorphic mutant. Here, we explored the specificity of the bam-Wolbachia interaction by generating 22 new bam mutants, with ten mutants displaying fertility defects. Nine of these mutants trend towards rescue by the w Mel Wolbachia variant, with eight statistically significant at the fertility and/or cytological level. In some cases, fertility was increased a striking 20-fold. There is no specificity between the rescue and the known binding regions of bam , suggesting w Mel does not interact with one singular bam partner to rescue the reproductive phenotype. We further tested if w Mel interacts with bam in a non-specific way, by increasing bam transcript levels or acting upstream in germline stem cells. A fertility assessment of a bam RNAi knockdown mutant reveals that w Mel rescue is specific to functionally mutant bam alleles and we find no obvious evidence of w Mel interaction with germline stem cells in bam mutants.

AUTHOR SUMMARY: Reproduction in the Drosophila melanogaster fruit fly is dependent on the bag of marbles (bam) gene, which acts early in the process of generating eggs and sperm. Mutations to this gene negatively impact the fertility of the fly, causing it to be sterile or have fewer progeny. Interestingly, we find that the bacteria Wolbachia , which resides within reproductive cells across a wide range of insects, partially restores the fertility and ovary phenotype of several bam mutants of which the resultant Bam protein is altered from wildtype. The protein function of Bam is further suggested to be important by the lack of rescue for a fly that has a fertility defect due to low expression of a non-mutated bam gene. Previous work makes similar conclusions about Wolbachia with another reproductive gene, Sex lethal (Sxl), highlighting the potential for rescue of fertility mutants to occur in a similar way across different genes. An understanding of the ways in which Wolbachia can affect host reproduction provides us with context with which to frame Wolbachia 's impact on host genes, such as bam and Sxl, and consider the evolutionary implications of Wolbachia 's infection in D. melanogaster fruit flies.}, } @article {pmid37630471, year = {2023}, author = {Chao, LL and Shih, CM}, title = {First Detection and Genetic Identification of Wolbachia Endosymbiont in Field-Caught Aedes aegypti (Diptera: Culicidae) Mosquitoes Collected from Southern Taiwan.}, journal = {Microorganisms}, volume = {11}, number = {8}, pages = {}, pmid = {37630471}, issn = {2076-2607}, support = {MOST 111-2314-B-037-031//Ministry of Science and Technology, Taiwan/ ; }, abstract = {The prevalence and genetic character of Wolbachia endosymbionts in field-collected Aedes aegypti mosquitoes were examined for the first time in Taiwan. A total of 665 Ae. aegypti were screened for Wolbachia infection using a PCR assay targeting the Wolbachia surface protein (wsp) gene. In general, the prevalence of Wolbachia infection was detected in 3.3% Ae. aegypti specimens (2.0% female and 5.2% male). Group-specific Wolbachia infection was detected with an infection rate of 1.8%, 0.8% and 0.8% in groups A, B and A&B, respectively. Genetic analysis demonstrated that all Wolbachia strains from Taiwan were phylogenetically affiliated with Wolbachia belonging to the supergroups A and B, with high sequence similarities of 99.4-100% and 99.2-100%, respectively. Phylogenetic relationships can be easily distinguished by maximum likelihood (ML) analysis and were congruent with the unweighted pair group with the arithmetic mean (UPGMA) method. The intra- and inter-group analysis of genetic distance (GD) values revealed a lower level within the Taiwan strains (GD < 0.006 for group A and GD < 0.008 for group B) and a higher level (GD > 0.498 for group A and GD > 0.286 for group B) as compared with other Wolbachia strains. Our results describe the first detection and molecular identification of Wolbachia endosymbiont in field-caught Ae. aegypti mosquitoes collected from Taiwan, and showed a low Wolbachia infection rate belonging to supergroups A and B in Ae. aegypti mosquitoes.}, } @article {pmid37628597, year = {2023}, author = {Corpuz, RL and Bellinger, MR and Veillet, A and Magnacca, KN and Price, DK}, title = {The Transmission Patterns of the Endosymbiont Wolbachia within the Hawaiian Drosophilidae Adaptive Radiation.}, journal = {Genes}, volume = {14}, number = {8}, pages = {}, pmid = {37628597}, issn = {2073-4425}, mesh = {Female ; Animals ; *Ecosystem ; Hawaii ; Phylogeny ; *Wolbachia/genetics ; Drosophila/genetics ; }, abstract = {The evolution of endosymbionts and their hosts can lead to highly dynamic interactions with varying fitness effects for both the endosymbiont and host species. Wolbachia, a ubiquitous endosymbiont of arthropods and nematodes, can have both beneficial and detrimental effects on host fitness. We documented the occurrence and patterns of transmission of Wolbachia within the Hawaiian Drosophilidae and examined the potential contributions of Wolbachia to the rapid diversification of their hosts. Screens for Wolbachia infections across a minimum of 140 species of Hawaiian Drosophila and Scaptomyza revealed species-level infections of 20.0%, and across all 399 samples, a general infection rate of 10.3%. Among the 44 Wolbachia strains we identified using a modified Wolbachia multi-locus strain typing scheme, 30 (68.18%) belonged to supergroup B, five (11.36%) belonged to supergroup A, and nine (20.45%) had alleles with conflicting supergroup assignments. Co-phylogenetic reconciliation analysis indicated that Wolbachia strain diversity within their endemic Hawaiian Drosophilidae hosts can be explained by vertical (e.g., co-speciation) and horizontal (e.g., host switch) modes of transmission. Results from stochastic character trait mapping suggest that horizontal transmission is associated with the preferred oviposition substrate of the host, but not the host's plant family or island of occurrence. For Hawaiian Drosophilid species of conservation concern, with 13 species listed as endangered and 1 listed as threatened, knowledge of Wolbachia strain types, infection status, and potential for superinfection could assist with conservation breeding programs designed to bolster population sizes, especially when wild populations are supplemented with laboratory-reared, translocated individuals. Future research aimed at improving the understanding of the mechanisms of Wolbachia transmission in nature, their impact on the host, and their role in host species formation may shed light on the influence of Wolbachia as an evolutionary driver, especially in Hawaiian ecosystems.}, } @article {pmid37622600, year = {2023}, author = {Ciocchetta, S and Frentiu, FD and Montarsi, F and Capelli, G and Devine, GJ}, title = {Investigation on key aspects of mating biology in the mosquito Aedes koreicus.}, journal = {Medical and veterinary entomology}, volume = {37}, number = {4}, pages = {826-833}, doi = {10.1111/mve.12687}, pmid = {37622600}, issn = {1365-2915}, mesh = {Female ; Male ; Animals ; *Aedes ; Reproduction ; Insemination ; Italy ; Biology ; Introduced Species ; Mosquito Vectors ; }, abstract = {Aedes koreicus Edwards, 1917 (Hulecoetomyia koreica) is a mosquito (Diptera: Culicidae) from Northeast Asia with a rapidly expanding presence outside its original native range. Over the years, the species has been discovered in several new countries, either spreading after first introduction or remaining localised to limited areas. Notably, recent studies have demonstrated the ability of the species to transmit zoonotic parasites and viruses both in the field and in laboratory settings. Combined with its invasive potential, the possible role of Ae. koreicus in pathogen transmission highlights the public health risks resulting from its invasion. In this study, we used a recently established population from Italy to investigate aspects of biology that influence reproductive success in Ae. koreicus: autogeny, mating behaviour, mating disruption by the sympatric invasive species Aedes albopictus Skuse, 1894, and the presence of the endosymbiont Wolbachia pipientis Hertig, 1936. Our laboratory population did not exhibit autogenic behaviour and required a bloodmeal to complete its ovarian cycle. When we exposed Ae. koreicus females to males of Ae. albopictus, we observed repeated attempts at insemination and an aggressive, disruptive mating behaviour initiated by male Ae. albopictus. Despite this, no sperm was identified in Ae. koreicus spermathecae. Wolbachia, an endosymbiotic bacterium capable of influencing mosquito reproductive behaviour, was not detected in this Ae. koreicus population and, therefore, had no effect on Ae. koreicus reproduction.}, } @article {pmid37612420, year = {2023}, author = {Strunov, A and Schoenherr, C and Kapun, M}, title = {Wolbachia has subtle effects on thermal preference in highly inbred Drosophila melanogaster which vary with life stage and environmental conditions.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {13792}, pmid = {37612420}, issn = {2045-2322}, mesh = {Animals ; Drosophila melanogaster ; *Wolbachia ; Drosophila ; *Arthropods ; Body Temperature ; }, abstract = {Temperature fluctuations are challenging for ectotherms which are not able to regulate body temperature by physiological means and thus have to adjust their thermal environment via behavior. However, little is yet known about whether microbial symbionts influence thermal preference (Tp) in ectotherms by modulating their physiology. Several recent studies have demonstrated substantial effects of Wolbachia infections on host Tp in different Drosophila species. These data indicate that the direction and strength of thermal preference variation is strongly dependent on host and symbiont genotypes and highly variable among studies. By employing highly controlled experiments, we investigated the impact of several environmental factors including humidity, food quality, light exposure, and experimental setup that may influence Tp measurements in adult Drosophila melanogaster flies. Additionally, we assessed the effects of Wolbachia infection on Tp of Drosophila at different developmental stages, which has not been done before. We find only subtle effects of Wolbachia on host Tp which are strongly affected by experimental variation in adult, but not during juvenile life stages. Our in-depth analyses show that environmental variation has a substantial influence on Tp which demonstrates the necessity of careful experimental design and cautious interpretations of Tp measurements together with a thorough description of the methods and equipment used to conduct behavioral studies.}, } @article {pmid37612118, year = {2023}, author = {Maruyama, J and Inoue, H and Hirose, Y and Nakabachi, A}, title = {16S rRNA Gene Sequencing of Six Psyllid Species of the Family Carsidaridae Identified Various Bacteria Including Symbiopectobacterium.}, journal = {Microbes and environments}, volume = {38}, number = {3}, pages = {}, pmid = {37612118}, issn = {1347-4405}, mesh = {Animals ; *Hemiptera ; RNA, Ribosomal, 16S/genetics ; Genes, rRNA ; Bacteria/genetics ; Enterobacteriaceae/genetics ; *Gammaproteobacteria/genetics ; }, abstract = {Psyllids (Hemiptera: Sternorrhyncha: Psylloidea) are plant sap-sucking insects that are closely associated with various microbes. To obtain a more detailed understanding of the ecological and evolutionary behaviors of microbes in Psylloidea, the bacterial populations of six psyllid species, belonging to the family Carsidaridae, were analyzed using high-throughput amplicon sequencing of the 16S rRNA gene. The majority of the secondary symbionts identified in the present study were gammaproteobacteria, particularly those of the order Enterobacterales, including Arsenophonus and Sodalis, which are lineages found in a wide variety of insect hosts. Additionally, Symbiopectobacterium, another Enterobacterales lineage, which has recently been recognized and increasingly shown to be vertically transmitted and mutualistic in various invertebrates, was identified for the first time in Psylloidea. This lineage is closely related to Pectobacterium spp., which are plant pathogens, but forms a distinct clade exhibiting no pathogenicity to plants. Non-Enterobacterales gammaproteobacteria found in the present study were Acinetobacter, Pseudomonas (both Pseudomonadales), Delftia, Comamonas (both Burkholderiales), and Xanthomonas (Xanthomonadales), a putative plant pathogen. Regarding alphaproteobacteria, three Wolbachia (Rickettsiales) lineages belonging to supergroup B, the major group in insect lineages, were detected in four psyllid species. In addition, a Wolbachia lineage of supergroup O, a minor group recently found for the first time in Psylloidea, was detected in one psyllid species. These results suggest the pervasive transfer of bacterial symbionts among animals and plants, providing deeper insights into the evolution of the interactions among these organisms.}, } @article {pmid37604924, year = {2023}, author = {Osorio, J and Villa-Arias, S and Camargo, C and Ramírez-Sánchez, LF and Barrientos, LM and Bedoya, C and Rúa-Uribe, G and Dorus, S and Alfonso-Parra, C and Avila, FW}, title = {wMel Wolbachia alters female post-mating behaviors and physiology in the dengue vector mosquito Aedes aegypti.}, journal = {Communications biology}, volume = {6}, number = {1}, pages = {865}, pmid = {37604924}, issn = {2399-3642}, mesh = {Animals ; Male ; Female ; Humans ; *Aedes ; Proteomics ; *Wolbachia ; Semen ; Mosquito Vectors ; *Dengue/prevention & control ; }, abstract = {Globally invasive Aedes aegypti disseminate numerous arboviruses that impact human health. One promising method to control Ae. aegypti populations is transinfection with Wolbachia pipientis, which naturally infects ~40-52% of insects but not Ae. aegypti. Transinfection of Ae. aegypti with the wMel Wolbachia strain induces cytoplasmic incompatibility (CI), allows infected individuals to invade native populations, and inhibits transmission of medically relevant arboviruses by females. Female insects undergo post-mating physiological and behavioral changes-referred to as the female post-mating response (PMR)-required for optimal fertility. PMRs are typically elicited by male seminal fluid proteins (SFPs) transferred with sperm during mating but can be modified by other factors, including microbiome composition. Wolbachia has modest effects on Ae. aegypti fertility, but its influence on other PMRs is unknown. Here, we show that Wolbachia influences female fecundity, fertility, and re-mating incidence and significantly extends the longevity of virgin females. Using proteomic methods to examine the seminal proteome of infected males, we found that Wolbachia moderately affects SFP composition. However, we identified 125 paternally transferred Wolbachia proteins, but the CI factor proteins (Cifs) were not among them. Our findings indicate that Wolbachia infection of Ae. aegypti alters female PMRs, potentially influencing control programs that utilize Wolbachia-infected individuals.}, } @article {pmid37603352, year = {2023}, author = {Zhu, Z and Hui, Y and Hu, L}, title = {The impact of predators of mosquito larvae on Wolbachia spreading dynamics.}, journal = {Journal of biological dynamics}, volume = {17}, number = {1}, pages = {2249024}, doi = {10.1080/17513758.2023.2249024}, pmid = {37603352}, issn = {1751-3766}, mesh = {Animals ; *Wolbachia ; Models, Biological ; Mosquito Vectors ; *Culicidae ; Larva ; }, abstract = {Dengue fever creates more than 390 million cases worldwide yearly. The most effective way to deal with this mosquito-borne disease is to control the vectors. In this work we consider two weapons, the endosymbiotic bacteria Wolbachia and predators of mosquito larvae, for combating the disease. As Wolbachia-infected mosquitoes are less able to transmit dengue virus, releasing infected mosquitoes to invade wild mosquito populations helps to reduce dengue transmission. Besides this measure, the introduction of predators of mosquito larvae can control mosquito population. To evaluate the impact of the predators on Wolbachia spreading dynamics, we develop a stage-structured five-dimensional model, which links the predator-prey dynamics with the Wolbachia spreading. By comparatively analysing the dynamics of the models without and with predators, we observe that the introduction of the predators augments the number of coexistence equilibria and impedes Wolbachia spreading. Some numerical simulations are presented to support and expand our theoretical results.}, } @article {pmid37602844, year = {2023}, author = {Cull, B and Wang, XR}, title = {Methods for the Study of Ticks, Mosquitoes, and their Transmitted Pathogens: Toward a Greater Understanding of Vector Biology and Arthropod-Microbe Interactions.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {193}, pages = {}, doi = {10.3791/64986}, pmid = {37602844}, issn = {1940-087X}, support = {R01 AI049424/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Arthropods ; Mosquito Vectors ; *Ixodes ; *Aedes ; Biology ; }, abstract = {Chen, L., Xiao, Q., Shi, M., Cheng, J., Wu, J. Detecting Wolbachia strain wAlbB in Aedes albopictus cell lines. Journal of Visualized Experiments. (184), e63662 (2022). Haziqah-Rashid, A. et al. Determining temperature preference of mosquitoes and other ectotherms. Journal of Visualized Experiments. (187), e64356 (2022). Huang, D. et al. Mosquito-associated virus isolation from field-collected mosquitoes. Journal of Visualized Experiments. (186), e63852 (2022). Khoo, B., Cull, B., Oliver, J. D. Tick artificial membrane feeding for Ixodes scapularis. Journal of Visualized Experiments. 64553 (2022). Leal-Galvan, B., Harvey, C., Thomas, D., Saelao, P., Oliva Chavez, A. S. Isolation of microRNAs from tick ex vivo salivary gland cultures and extracellular vesicles. Journal of Visualized Experiments. (182), e63618 (2022). Liang, Q. et al. Control of Aedes albopictus mosquito larvae with Carpesium abrotanoides L. Journal of Visualized Experiments. (186), e63976 (2022). Wang, F. et al. Experimental viral infection in adult mosquitoes by oral feeding and microinjection. Journal of Visualized Experiments. (185), e63830 (2022). Wang, X. R., Burkhardt, N. Y., Price, L. D., Munderloh, U. G. An electroporation method to transform Rickettsia spp. with a fluorescent protein-expressing shuttle vector in tick cell lines. Journal of Visualized Experiments. (188), e64562 (2022).}, } @article {pmid37601442, year = {2023}, author = {Archer, J and Hurst, GDD and Hornett, EA}, title = {Male-killer symbiont screening reveals novel associations in Adalia ladybirds.}, journal = {Access microbiology}, volume = {5}, number = {7}, pages = {}, pmid = {37601442}, issn = {2516-8290}, abstract = {While male-killing bacteria are known to infect across arthropods, ladybird beetles represent a hotspot for these symbioses. In some host species, there are multiple different symbionts that vary in presence and frequency between populations. To further our understanding of spatial and frequency variation, we tested for the presence of three male-killing bacteria: Wolbachia , Rickettsia and Spiroplasma , in two Adalia ladybird species from a previously unexplored UK population. The two-spot ladybird, A. bipunctata, is known to harbour all three male-killers, and we identified Spiroplasma infection in the Merseyside population for the first time. However, in contrast to previous studies on two-spot ladybirds from continental Europe, evidence from egg-hatch rates indicates the Spiroplasma strain present in the Merseyside population does not cause embryonic male-killing. In the related ten-spot ladybird, A. decempunctata, there is only one previous record of a male-killing symbiont, a Rickettsia , which we did not detect in the Merseyside sample. However, PCR assays indicated the presence of a Spiroplasma in a single A. decempunctata specimen. Marker sequence indicated that this Spiroplasma was divergent from that found in sympatric A. bipunctata. Genome sequencing of the Spiroplasma -infected A. decempunctata additionally revealed the presence of cobionts in the form of a Centistes parasitoid wasp and the parasitic fungi Beauveria. Further study of A. decempunctata from this population is needed to resolve whether it is the ladybird or wasp cobiont that harbours Spiroplasma , and to establish the phenotype of this strain. These data indicate first that microbial symbiont phenotype should not be assumed from past studies conducted in different locations, and second that cobiont presence may confound screening studies aimed to detect the frequency of a symbiont in field collected material from a focal host species.}, } @article {pmid37584011, year = {2023}, author = {Perles, L and Otranto, D and Barreto, WTG and de Macedo, GC and Lia, RP and Mendoza-Roldan, JA and Herrera, HM and de Oliveira, CE and Machado, RZ and André, MR}, title = {Mansonella sp. and associated Wolbachia endosymbionts in ring-tailed coatis (Nasua nasua) in periurban areas from Midwestern Brazil.}, journal = {International journal for parasitology. Parasites and wildlife}, volume = {22}, number = {}, pages = {14-19}, pmid = {37584011}, issn = {2213-2244}, abstract = {Coatis (Nasua nasua) are wild carnivorous well adapted to anthropized environments especially important because they act as reservoirs hosts for many arthropod-borne zoonotic pathogens. Information about filarioids from coatis and associated Wolbachia spp. in Brazil is scant. To investigate the diversity of filarial nematodes, blood samples (n = 100 animals) were obtained from two urban areas in midwestern Brazil and analyzed using blood smears and buffy coats and cPCR assays based on the cox1, 12S rRNA, 18S rRNA, hsp70 and myoHC genes for nematodes and 16S rRNA for Wolbachia. When analyzing coati blood smears and buffy coats, 30% and 80% of the samples presented at least one microfilaria, respectively. Twenty-five cox1 sequences were obtained showing 89% nucleotide identity with Mansonella ozzardi. Phylogenetic analyses clustered cox1 sequences herein obtained within the Mansonella spp. clade. Sequences of both myoHC and two hsp70 genes showed 99.8% nucleotide identity with Mansonella sp. and clustered into a clade within Mansonella sp., previously detected in coatis from Brazil. Two blood samples were positive for Wolbachia, with a 99% nucleotide identity with Wolbachia previously found in Mansonella perstans, Mansonella ozzardi and Mansonella atelensis and in ectoparasites of the genus Pseudolynchia, Melophagus and Cimex. The study showed a high prevalence of Mansonella sp. in the coati population examined, suggesting that this animal species play a role as reservoirs of a novel, yet to be described, species within the Onchocercidae family.}, } @article {pmid37577425, year = {2023}, author = {Garrido, M and Veiga, J and Garrigós, M and Martínez-de la Puente, J}, title = {The interplay between vector microbial community and pathogen transmission on the invasive Asian tiger mosquito, Aedes albopictus: current knowledge and future directions.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1208633}, pmid = {37577425}, issn = {1664-302X}, abstract = {The invasive Asian tiger mosquito Aedes albopictus is nowadays broadly distributed with established populations in all continents except Antarctica. In the invaded areas, this species represents an important nuisance for humans and, more relevant, it is involved in the local transmission of pathogens relevant under a public health perspective. Aedes albopictus is a competent vector of parasites such as Dirofilaria and viruses including dengue virus, Zika virus, and chikungunya virus, among others. The mosquito microbiota has been identified as one of the major drivers of vector competence, acting upon relevant vector functions as development or immunity. Here, we review the available literature on the interaction between Ae. albopictus microbiota and pathogen transmission and identify the knowledge gaps on the topic. Most studies are strictly focused on the interplay between pathogens and Wolbachia endosymbiont while studies screening whole microbiota are still scarce but increasing in recent years, supported on Next-generation sequencing tools. Most experimental trials use lab-reared mosquitoes or cell lines, exploring the molecular mechanisms of the microbiota-pathogen interaction. Yet, correlational studies on wild populations are underrepresented. Consequently, we still lack sufficient evidence to reveal whether the microbiota of introduced populations of Ae. albopictus differ from those of native populations, or how microbiota is shaped by different environmental and anthropic factors, but especially, how these changes affect the ability of Ae. albopictus to transmit pathogens and favor the occurrence of outbreaks in the colonized areas. Finally, we propose future research directions on this research topic.}, } @article {pmid37567493, year = {2023}, author = {Kamkong, P and Jitsamai, W and Thongmeesee, K and Ratthawongjirakul, P and Taweethavonsawat, P}, title = {Genetic diversity and characterization of Wolbachia endosymbiont in canine filariasis.}, journal = {Acta tropica}, volume = {246}, number = {}, pages = {107000}, doi = {10.1016/j.actatropica.2023.107000}, pmid = {37567493}, issn = {1873-6254}, mesh = {Animals ; Dogs ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; *Dirofilariasis ; *Dirofilaria immitis/genetics ; Dirofilaria ; *Filarioidea/genetics ; *Heart Diseases ; Genetic Variation ; *Dog Diseases ; }, abstract = {Canine filariasis is caused by nematodes from the family Onchocercidae, which is transmitted by arthropod vectors. The disease is commonly found in Southeast Asia and exists worldwide. Some filarial nematodes are associated with intracellular bacteria of the genus Wolbachia, which plays an important role in embryogenesis, molting, and the long-term survival of adult worms. This study aims to characterize Wolbachia sp. and determine the association between Wolbachia and canine filarial nematode species in Thailand. A total of 46 dog blood samples that were naturally infected with filarial nematodes were obtained to identify filarial nematode species by Giemsa stained under a light microscope and confirmed using the molecular technique. In order to characterize Wolbachia sp., the nested PCR assay targeting the 16S rRNA gene showed that all samples of Dirofilaria immitis and fifteen samples of Candidatus Dirofilaria hongkongensis were grouped into Wolbachia supergroup C. In addition, all samples of Brugia spp. and five samples of Candidatus Dirofilaria hongkongensis were classified into Wolbachia supergroup D. The genetic diversity analysis conducted using the 16S rRNA gene revealed a similar result when analyzed through phylogenetic tree analysis. This is the first genetic diversity study of Wolbachia of Candidatus Dirofilaria hongkongensis in infected dogs in Thailand.}, } @article {pmid37546688, year = {2023}, author = {Vásquez, VN and Kueppers, LM and Rašić, G and Marshall, JM}, title = {wMel replacement of dengue-competent mosquitoes is robust to near-term change.}, journal = {Nature climate change}, volume = {13}, number = {8}, pages = {848-855}, pmid = {37546688}, issn = {1758-678X}, support = {R01 AI143698/AI/NIAID NIH HHS/United States ; }, abstract = {Rising temperatures are impacting the range and prevalence of mosquito-borne diseases. A promising biocontrol technology replaces wild mosquitoes with those carrying the virus-blocking Wolbachia bacterium. Because the most widely used strain, wMel, is adversely affected by heat stress, we examined how global warming may influence wMel-based replacement. We simulated interventions in two locations with successful field trials using Coupled Model Intercomparison Project Phase 5 climate projections and historical temperature records, integrating empirical data on wMel's thermal sensitivity into a model of Aedes aegypti population dynamics to evaluate introgression and persistence over one year. We show that in Cairns, Australia, climatic futures necessitate operational adaptations for heatwaves exceeding two weeks. In Nha Trang, Vietnam, projected heatwaves of three weeks and longer eliminate wMel under the most stringent assumptions of that symbiont's thermal limits. We conclude that this technology is generally robust to near-term (2030s) climate change. Accelerated warming may challenge this in the 2050s and beyond.}, } @article {pmid37533804, year = {2023}, author = {Namias, A and Sahlin, K and Makoundou, P and Bonnici, I and Sicard, M and Belkhir, K and Weill, M}, title = {Nanopore sequencing of PCR products enables multicopy gene family reconstruction.}, journal = {Computational and structural biotechnology journal}, volume = {21}, number = {}, pages = {3656-3664}, pmid = {37533804}, issn = {2001-0370}, abstract = {The importance of gene amplifications in evolution is more and more recognized. Yet, tools to study multi-copy gene families are still scarce, and many such families are overlooked using common sequencing methods. Haplotype reconstruction is even harder for polymorphic multi-copy gene families. Here, we show that all variants (or haplotypes) of a multi-copy gene family present in a single genome, can be obtained using Oxford Nanopore Technologies sequencing of PCR products, followed by steps of mapping, SNP calling and haplotyping. As a proof of concept, we acquired the sequences of highly similar variants of the cidA and cidB genes present in the genome of the Wolbachia wPip, a bacterium infecting Culex pipiens mosquitoes. Our method relies on a wide database of cid genes, previously acquired by cloning and Sanger sequencing. We addressed problems commonly faced when using mapping approaches for multi-copy gene families with highly similar variants. In addition, we confirmed that PCR amplification causes frequent chimeras which have to be carefully considered when working on families of recombinant genes. We tested the robustness of the method using a combination of bioinformatics (read simulations) and molecular biology approaches (sequence acquisitions through cloning and Sanger sequencing, specific PCRs and digital droplet PCR). When different haplotypes present within a single genome cannot be reconstructed from short reads sequencing, this pipeline confers a high throughput acquisition, gives reliable results as well as insights of the relative copy numbers of the different variants.}, } @article {pmid37532686, year = {2023}, author = {Hasnaoui, B and Bérenger, JM and Delaunay, P and Diarra, AZ and Ndiaye, EHI and M'madi, SA and Masotti, N and Sevestre, J and Parola, P}, title = {Survey of bed bug infestations in homeless shelters in southern France.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {12557}, pmid = {37532686}, issn = {2045-2322}, mesh = {Animals ; Humans ; *Bedbugs ; Surveys and Questionnaires ; Public Health ; *Ectoparasitic Infestations ; France ; *Ill-Housed Persons ; }, abstract = {Bed bug has become a major public health pest worldwide. Infestation may result in numerous negative health effects. Homeless shelters are one of the most habitats that can be infested with bed bugs, a few studies have focused on bed bug infestations in these settings. We conducted a survey of infestations of bed bugs in a homeless shelter in southern France, using an innovative seven-level scale (0-6) to assess the degree of infestation, MALDI TOF-MS to identify bed bugs, and a biomolecular tool to detect bacteria. Bed bug infestations were documented in 13% (9/68) of investigated rooms. A total of 184 bed bugs were collected and morphologically identified as Cimex lectularius. MALDI TOF-MS analysis allowed us to obtain high-quality MS spectra for all 184 specimens, to correctly identify all specimens, and included 178/184 (97%) Log Score Values higher than 1.8. Among the bacteria tested, Wolbachia sp. DNA was found in 149/184 (81%) of the bed bugs, and one sample was positive for Coxiella burnetii, the agent of Q fever. Our study is the first of its kind that offers new perspectives for increasing public awareness of the conditions in homeless shelters.}, } @article {pmid37525959, year = {2023}, author = {Kolasa, M and Kajtoch, Ł and Michalik, A and Maryańska-Nadachowska, A and Łukasik, P}, title = {Till evolution do us part: The diversity of symbiotic associations across populations of Philaenus spittlebugs.}, journal = {Environmental microbiology}, volume = {25}, number = {11}, pages = {2431-2446}, doi = {10.1111/1462-2920.16473}, pmid = {37525959}, issn = {1462-2920}, mesh = {Humans ; Animals ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; Enterobacteriaceae/genetics ; Bacteria/genetics ; *Hemiptera/microbiology ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Symbiotic bacteria have played crucial roles in the evolution of sap-feeding insects and can strongly affect host function. However, their diversity and distribution within species are not well understood; we do not know to what extent environmental factors or associations with other species may affect microbial community profiles. We addressed this question in Philaenus spittlebugs by surveying both insect and bacterial marker gene amplicons across multiple host populations. Host mitochondrial sequence data confirmed morphology-based identification of six species and revealed two divergent clades of Philaenus spumarius. All of them hosted the primary symbiont Sulcia that was almost always accompanied by Sodalis. Interestingly, populations and individuals often differed in the presence of Sodalis sequence variants, suggestive of intra-genome 16S rRNA variant polymorphism combined with rapid genome evolution and/or recent additional infections or replacements of the co-primary symbiont. The prevalence of facultative endosymbionts, including Wolbachia, Rickettsia, and Spiroplasma, varied among populations. Notably, cytochrome I oxidase (COI) amplicon data also showed that nearly a quarter of P. spumarius were infected by parasitoid flies (Verralia aucta). One of the Wolbachia operational taxonomic units (OTUs) was exclusively present in Verralia-parasitized specimens, suggestive of parasitoids as their source and highlighting the utility of host gene amplicon sequencing in microbiome studies.}, } @article {pmid37524915, year = {2023}, author = {Lee, H and Seo, P and Teklay, S and Yuguchi, E and Benetta, ED and Werren, JH and Ferree, PM}, title = {Ability of a selfish B chromosome to evade genome elimination in the jewel wasp, Nasonia vitripennis.}, journal = {Heredity}, volume = {131}, number = {3}, pages = {230-237}, pmid = {37524915}, issn = {1365-2540}, mesh = {*Genome, Insect ; Animals ; *Chromosomes, Insect ; Protamines/genetics/metabolism ; Histones/genetics/metabolism ; Male ; Female ; Genes, rRNA ; DNA, Ribosomal/genetics/metabolism ; Genetic Loci ; }, abstract = {B chromosomes are non-essential, extra chromosomes that can exhibit transmission-enhancing behaviors, including meiotic drive, mitotic drive, and induction of genome elimination, in plants and animals. A fundamental but poorly understood question is what characteristics allow B chromosomes to exhibit these extraordinary behaviors. The jewel wasp, Nasonia vitripennis, harbors a heterochromatic, paternally transmitted B chromosome known as paternal sex ratio (PSR), which causes complete elimination of the sperm-contributed half of the genome during the first mitotic division of fertilized embryos. This genome elimination event may result from specific, previously observed alterations of the paternal chromatin. Due to the haplo-diploid reproduction of the wasp, genome elimination by PSR causes female-destined embryos to develop as haploid males that transmit PSR. PSR does not undergo self-elimination despite its presence with the paternal chromatin until the elimination event. Here we performed fluorescence microscopic analyses aimed at understanding this unexplained property. Our results show that PSR, like the rest of the genome, participates in the histone-to-protamine transition, arguing that PSR does not avoid this transition to escape self-elimination. In addition, PSR partially escapes the chromatin-altering activity of the intracellular bacterium, Wolbachia, demonstrating that this ability to evade chromatin alteration is not limited to PSR's own activity. Finally, we observed that the rDNA locus and other unidentified heterochromatic regions of the wasp's genome also seem to evade chromatin disruption by PSR, suggesting that PSR's genome-eliminating activity does not affect heterochromatin. Thus, PSR may target an aspect of euchromatin to cause genome elimination.}, } @article {pmid37491306, year = {2023}, author = {Moorhead, AR and Evans, CC and Sakamoto, K and Dzimianski, MT and Mansour, A and DiCosty, U and Fricks, C and McCall, S and Carson, B and Nelson, CT and McCall, JW}, title = {Effects of doxycycline dose rate and pre-adulticide wait period on heartworm-associated pathology and adult worm mass.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {251}, pmid = {37491306}, issn = {1756-3305}, mesh = {Animals ; Dogs ; *Dirofilaria immitis ; Doxycycline ; *Dirofilariasis/drug therapy ; *Filaricides ; *Dog Diseases/drug therapy ; *Wolbachia ; }, abstract = {BACKGROUND: The American Heartworm Society canine guidelines recommend treatment with doxycycline prior to adulticide administration to reduce levels of Wolbachia and its associated metabolites, which are known to be a leading cause of pulmonary pathology. Studies have determined that doxycycline administered at 10 mg/kg BID for 28 days is an effective dose for eliminating Wolbachia, but what has not been determined is the clinical relevance of this elimination. The current guidelines also recommend a 30-day wait period following administration of doxycycline to allow for clearance of metabolites, such as Wolbachia surface protein, and for further reduction in heartworm biomass before administration of adulticide. Reducing the doxycycline dose and eliminating the wait period may carry practical benefits for the animal, client, and practitioner.

METHODS: To investigate these treatment practices, Dirofilaria immitis adults were surgically transplanted into each of 45 dogs, which were divided into nine study groups of five dogs each. Seventy-five days after transplantation, two groups each were administered 5, 7.5, or 10 mg/kg BID doxycycline orally for 28 days and 6 µg/kg ivermectin monthly, with three untreated groups serving as controls. Study animals were necropsied and examined prior to treatment as well as 30 and 60 days post-treatment.

RESULTS: Mean worm weight was unaffected by dosage but exhibited a significant increase at 30 days and significant decrease at 60 days post-treatment, including in control groups. Histopathology lesion scores did not significantly differ among groups, with the exception of the lung composite score for one untreated group. Liver enzymes, the levels of which are a concern in doxycycline treatment, were also examined, with no abnormalities in alanine aminotransferase or alkaline phosphatase observed.

CONCLUSIONS: No consistent worsening of tissue lesions was observed with or without the AHS-recommended 30-day wait period, nor did reduced dosages of doxycycline lead to worsening of pathology or any change in efficacy in depleting worm weight. Mean worm weight did significantly increase prior to, and decrease following, the wait period. Future work that also includes adulticide treatment (i.e. melarsomine) will study treatment recommendations that may improve both animal health and owner compliance.}, } @article {pmid37484687, year = {2023}, author = {Adams, GJ and O'Brien, PA}, title = {The unified theory of sleep: Eukaryotes endosymbiotic relationship with mitochondria and REM the push-back response for awakening.}, journal = {Neurobiology of sleep and circadian rhythms}, volume = {15}, number = {}, pages = {100100}, pmid = {37484687}, issn = {2451-9944}, abstract = {The Unified Theory suggests that sleep is a process that developed in eukaryotic animals from a relationship with an endosymbiotic bacterium. Over evolutionary time the bacterium evolved into the modern mitochondrion that continues to exert an effect on sleep patterns, e.g. the bacterium Wolbachia establishes an endosymbiotic relationship with Drosophila and many other species of insects and is able to change the host's behaviour by making it sleep. The hypothesis is supported by other host-parasite relationships, e.g., Trypanosoma brucei which causes day-time sleepiness and night-time insomnia in humans and cattle. For eukaryotes such as Monocercomonoids that don't contain mitochondria we find no evidence of them sleeping. Mitochondria produce the neurotransmitter gamma aminobutyric acid (GABA), and ornithine a precursor of the neurotransmitter GABA, together with substances such as 3,4dihydroxy phenylalanine (DOPA) a precursor for the neurotransmitter dopamine: These substances have been shown to affect the sleep/wake cycles in animals such as Drosophilia and Hydra. Eukaryote animals have traded the very positive side of having mitochondria providing aerobic respiration for them with the negative side of having to sleep. NREM (Quiet sleep) is the process endosymbionts have imposed upon their host eukaryotes and REM (Active sleep) is the push-back adaptation of eukaryotes with brains, returning to wakefulness.}, } @article {pmid37477269, year = {2023}, author = {Ferreira, MU and Crainey, JL and Gobbi, FG}, title = {The search for better treatment strategies for mansonellosis: an expert perspective.}, journal = {Expert opinion on pharmacotherapy}, volume = {24}, number = {15}, pages = {1685-1692}, doi = {10.1080/14656566.2023.2240235}, pmid = {37477269}, issn = {1744-7666}, mesh = {Adult ; Animals ; Humans ; *Mansonelliasis/complications/drug therapy ; Mansonella ; Ivermectin/therapeutic use ; Anti-Bacterial Agents/therapeutic use ; *Anthelmintics/therapeutic use ; Arthralgia/complications/drug therapy ; }, abstract = {INTRODUCTION: Four species of the Mansonella genus infect millions of people across sub-Saharan Africa and Central and South America. Most infections are asymptomatic, but mansonellosis can be associated with nonspecific clinical manifestations such as fever, headache, arthralgia, and ocular lesions (M. ozzardi); pruritus, arthralgia, abdominal pain, angioedema, skin rash, and fatigue (M. perstans and perhaps Mansonella sp. 'DEUX'); and pruritic dermatitis and chronic lymphadenitis (M. perstans).

AREAS COVERED: We searched the PubMed and SciELO databases for publications on mansonelliasis in English, Spanish, Portuguese, or French that appeared until 1 May 2023. Literature data show that anthelmintics - single-dose ivermectin for M. ozzardi, repeated doses of mebendazole alone or in combination with diethylcarbamazine (DEC) for M. perstans, and DEC alone for M. streptocerca - are effective against microfilariae. Antibiotics that target Wolbachia endosymbionts, such as doxycycline, are likely to kill adult worms of most, if not all, Mansonella species, but the currently recommended 6-week regimen is relatively impractical. New anthelmintics and shorter antibiotic regimens (e.g. with rifampin) have shown promise in experimental filarial infections and may proceed to clinical trials.

EXPERT OPINION: We recommend that human infections with Mansonella species be treated, regardless of any apparent clinical manifestations. We argue that mansonellosis, despite being widely considered a benign infection, may represent a direct or indirect cause of significant morbidity that remains poorly characterized at present.}, } @article {pmid37474590, year = {2023}, author = {Rainey, SM and Geoghegan, V and Lefteri, DA and Ant, TH and Martinez, J and McNamara, CJ and Kamel, W and de Laurent, ZR and Castello, A and Sinkins, SP}, title = {Differences in proteome perturbations caused by the Wolbachia strain wAu suggest multiple mechanisms of Wolbachia-mediated antiviral activity.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {11737}, pmid = {37474590}, issn = {2045-2322}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; 202888/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Dengue Virus/physiology ; Proteome ; *Wolbachia/physiology ; Antiviral Agents ; Proteomics ; Lipids ; *Aedes ; }, abstract = {Some strains of the inherited bacterium Wolbachia have been shown to be effective at reducing the transmission of dengue virus (DENV) and other RNA viruses by Aedes aegypti in both laboratory and field settings and are being deployed for DENV control. The degree of virus inhibition varies between Wolbachia strains. Density and tissue tropism can contribute to these differences but there are also indications that this is not the only factor involved: for example, strains wAu and wAlbA are maintained at similar intracellular densities but only wAu produces strong DENV inhibition. We previously reported perturbations in lipid transport dynamics, including sequestration of cholesterol in lipid droplets, with strains wMel/wMelPop in Ae. aegypti. To further investigate the cellular basis underlying these differences, proteomic analysis of midguts was carried out on Ae. aegypti lines carrying strains wAu and wAlbA: with the hypothesis that differences in perturbations may underline Wolbachia-mediated antiviral activity. Surprisingly, wAu-carrying midguts not only showed distinct proteome perturbations when compared to non-Wolbachia carrying and wAlbA-carrying midguts but also wMel-carrying midguts. There are changes in RNA processing pathways and upregulation of a specific set of RNA-binding proteins in the wAu-carrying line, including genes with known antiviral activity. Lipid transport and metabolism proteome changes also differ between strains, and we show that strain wAu does not produce the same cholesterol sequestration phenotype as wMel. Moreover, in contrast to wMel, wAu antiviral activity was not rescued by cyclodextrin treatment. Together these results suggest that wAu could show unique features in its inhibition of arboviruses compared to previously characterized Wolbachia strains.}, } @article {pmid37464886, year = {2023}, author = {Shen, R and Wenzel, M and Messer, PW and Aquadro, CF}, title = {Evolution under a model of functionally buffered deleterious mutations can lead to positive selection in protein-coding genes.}, journal = {Evolution; international journal of organic evolution}, volume = {77}, number = {10}, pages = {2200-2212}, pmid = {37464886}, issn = {1558-5646}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; Evolution, Molecular ; Drosophila/genetics ; Selection, Genetic ; Mutation ; Amino Acids/genetics ; RNA-Binding Proteins/genetics ; *Drosophila Proteins/genetics ; }, abstract = {Selective pressures on DNA sequences often result in departures from neutral evolution that can be captured by the McDonald-Kreitman (MK) test. However, the nature of such selective forces often remains unknown to experimentalists. Amino acid fixations driven by natural selection in protein-coding genes are commonly associated with a genetic arms race or changing biological purposes, leading to proteins with new functionality. Here, we evaluate the expectations of population genetic patterns under a buffering mechanism driving selective amino acids to fixation, which is motivated by an observed phenotypic rescue of otherwise deleterious nonsynonymous substitutions at bag of marbles (bam) and Sex lethal (Sxl) in Drosophila melanogaster. These two genes were shown to experience strong episodic bursts of natural selection potentially due to infections of the endosymbiotic bacteria Wolbachia observed among multiple Drosophila species. Using simulations to implement and evaluate the evolutionary dynamics of a Wolbachia buffering model, we demonstrate that selectively fixed amino acid replacements will occur, but that the proportion of adaptive amino acid fixations and the statistical power of the MK test to detect the departure from an equilibrium neutral model are both significantly lower than seen for an arms race/change-in-function model that favors proteins with diversified amino acids. We find that the observed selection pattern at bam in a natural population of D. melanogaster is more consistent with an arms race model than with the buffering model.}, } @article {pmid37443389, year = {2023}, author = {Wybouw, N and Van Reempts, E and Zarka, J and Zélé, F and Bonte, D}, title = {Egg provisioning explains the penetrance of symbiont-mediated sex allocation distortion in haplodiploids.}, journal = {Heredity}, volume = {131}, number = {3}, pages = {221-229}, pmid = {37443389}, issn = {1365-2540}, mesh = {Animals ; Male ; Female ; Reproduction/physiology ; Penetrance ; *Tetranychidae/genetics ; Bacteroidetes ; Cytoplasm ; *Wolbachia/genetics ; Symbiosis/genetics ; Sex Ratio ; }, abstract = {Maternally transmitted symbionts such as Wolbachia can alter sex allocation in haplodiploid arthropods. By biasing population sex ratios towards females, these changes in sex allocation may facilitate the spread of symbionts. In contrast to symbiont-induced cytoplasmic incompatibility (CI), the mechanisms that underpin sex allocation distortion remain poorly understood. Using a nuclear genotype reference panel of the haplodiploid mite Tetranychus urticae and a single Wolbachia variant that is able to simultaneously induce sex allocation distortion and CI, we unraveled the mechanistic basis of Wolbachia-mediated sex allocation distortion. Host genotype was an important determinant for the strength of sex allocation distortion. We further show that sex allocation distortion by Wolbachia in haplodiploid mites is driven by increasing egg size, hereby promoting egg fertilization. This change in reproductive physiology was also coupled to increased male and female adult size. Our results echo previous work on Cardinium symbionts, suggesting that sex allocation distortion by regulating host investment in egg size is a common strategy among symbionts that infect haplodiploids. To better understand the relevance that sex allocation distortion may have for the spread of Wolbachia in natural haplodiploid populations, we parametrized a model based on generated phenotypic data. Our simulations show that empirically derived levels of sex allocation distortion can be sufficient to remove invasion thresholds, allowing CI to drive the spread of Wolbachia independently of the initial infection frequency. Our findings help elucidate the mechanisms that underlie the widespread occurrence of symbionts in haplodiploid arthropods and the evolution of sex allocation.}, } @article {pmid37432953, year = {2023}, author = {Medina, P and Russell, SL and Corbett-Detig, R}, title = {Deep data mining reveals variable abundance and distribution of microbial reproductive manipulators within and among diverse host species.}, journal = {PloS one}, volume = {18}, number = {7}, pages = {e0288261}, pmid = {37432953}, issn = {1932-6203}, support = {R35 GM128932/GM/NIGMS NIH HHS/United States ; T32 HG008345/HG/NHGRI NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/genetics ; *Arthropods ; Data Mining ; Ecology ; Reproduction ; *Wolbachia/genetics ; }, abstract = {Bacterial symbionts that manipulate the reproduction of their hosts are important factors in invertebrate ecology and evolution, and are being leveraged for host biological control. Infection prevalence restricts which biological control strategies are possible and is thought to be strongly influenced by the density of symbiont infection within hosts, termed titer. Current methods to estimate infection prevalence and symbiont titers are low-throughput, biased towards sampling infected species, and rarely measure titer. Here we develop a data mining approach to estimate symbiont infection frequencies within host species and titers within host tissues. We applied this approach to screen ~32,000 publicly available sequence samples from the most common symbiont host taxa, discovering 2,083 arthropod and 119 nematode infected samples. From these data, we estimated that Wolbachia infects approximately 44% of all arthropod and 34% of all nematode species, while other reproductive manipulators only infect 1-8% of arthropod and nematode species. Although relative titers within hosts were highly variable within and between arthropod species, a combination of arthropod host species and Wolbachia strain explained approximately 36% of variation in Wolbachia titer across the dataset. To explore potential mechanisms for host control of symbiont titer, we leveraged population genomic data from the model system Drosophila melanogaster. In this host, we found a number of SNPs associated with titer in candidate genes potentially relevant to host interactions with Wolbachia. Our study demonstrates that data mining is a powerful tool to detect bacterial infections and quantify infection intensities, thus opening an array of previously inaccessible data for further analysis in host-symbiont evolution.}, } @article {pmid37430172, year = {2023}, author = {Porter, J and Sullivan, W}, title = {The cellular lives of Wolbachia.}, journal = {Nature reviews. Microbiology}, volume = {21}, number = {11}, pages = {750-766}, pmid = {37430172}, issn = {1740-1534}, abstract = {Wolbachia are successful Gram-negative bacterial endosymbionts, globally infecting a large fraction of arthropod species and filarial nematodes. Efficient vertical transmission, the capacity for horizontal transmission, manipulation of host reproduction and enhancement of host fitness can promote the spread both within and between species. Wolbachia are abundant and can occupy extraordinary diverse and evolutionary distant host species, suggesting that they have evolved to engage and manipulate highly conserved core cellular processes. Here, we review recent studies identifying Wolbachia-host interactions at the molecular and cellular levels. We explore how Wolbachia interact with a wide array of host cytoplasmic and nuclear components in order to thrive in a diversity of cell types and cellular environments. This endosymbiont has also evolved the ability to precisely target and manipulate specific phases of the host cell cycle. The remarkable diversity of cellular interactions distinguishes Wolbachia from other endosymbionts and is largely responsible for facilitating its global propagation through host populations. Finally, we describe how insights into Wolbachia-host cellular interactions have led to promising applications in controlling insect-borne and filarial nematode-based diseases.}, } @article {pmid37428804, year = {2023}, author = {Alami, N and Carter, DC and Kwatra, NV and Zhao, W and Snodgrass, L and Porcalla, AR and Klein, CE and Cohen, DE and Gallenberg, L and Neenan, M and Carr, RA and Marsh, KC and Kempf, DJ}, title = {A Phase-I pharmacokinetic, safety and food-effect study on flubentylosin, a novel analog of Tylosin-A having potent anti-Wolbachia and antifilarial activity.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {7}, pages = {e0011392}, pmid = {37428804}, issn = {1935-2735}, mesh = {Adult ; Humans ; Tylosin ; *Wolbachia ; *Onchocerciasis ; Double-Blind Method ; Anti-Bacterial Agents/pharmacokinetics ; Macrolides ; Area Under Curve ; Dose-Response Relationship, Drug ; Administration, Oral ; }, abstract = {BACKGROUND: The parasitic filariae responsible for onchocerciasis and lymphatic filariasis are host to an endosymbiotic bacterium, Wolbachia, which is essential to the fertility and development of the parasites. We performed a Phase-I pharmacokinetic, safety and food-effect study on single and multiple ascending doses of flubentylosin (ABBV-4083), a macrolide antibacterial with activity against Wolbachia, intended to sterilize and eliminate the parasites.

METHODS: Seventy-eight healthy adults were exposed to flubentylosin; 36 were exposed to single ascending 40, 100, 200, 400 or 1000 mg doses; 12 received 1000 mg in the food-effect part; and 30 received multiple ascending daily doses of 100 mg for 7 days, 200 mg for 7 or 14 days, or 400 mg for 7 or 14 days. Twenty-two subjects received placebo.

RESULTS: Maximum concentrations (Cmax) of flubentylosin were reached after 1-2 hours, with a half-life < 4 hours at doses ≤ 400 mg. Cmax and AUC increased in a more than dose-proportional manner, with similar exposure after multiple dose administration. The most frequently reported adverse events were nausea (8/78, 10%) and headache (6/78, 8%). Two subjects given a single dose of flubentylosin 1000 mg in the food-effect part experienced reversible asymptomatic ALT and AST elevations at Grade 2 or Grade 4, with no elevation in bilirubin, deemed related to study drug. The effect of food on exposure parameters was minimal. No treatment-related serious adverse events were reported.

DISCUSSION: Flubentylosin 400 mg for 14 days was the maximum tolerated dose in this first-in-human, Phase-I study in healthy adults. Based on preclinical pharmacokinetic/pharmacodynamic modeling, flubentylosin 400 mg once daily for 7 or 14 days is expected to be an effective dose. A Phase-II, proof-of-concept study with flubentylosin using these regimens is currently ongoing in patients with onchocerciasis in Africa.}, } @article {pmid37426014, year = {2023}, author = {Marriott, AE and Dagley, JL and Hegde, S and Steven, A and Fricks, C and DiCosty, U and Mansour, A and Campbell, EJ and Wilson, CM and Gusovsky, F and Ward, SA and Hong, WD and O'Neill, P and Moorhead, A and McCall, S and McCall, JW and Taylor, MJ and Turner, JD}, title = {Dirofilariasis mouse models for heartworm preclinical research.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1208301}, pmid = {37426014}, issn = {1664-302X}, abstract = {INTRODUCTION: Dirofilariasis, including heartworm disease, is a major emergent veterinary parasitic infection and a human zoonosis. Currently, experimental infections of cats and dogs are used in veterinary heartworm preclinical drug research.

METHODS: As a refined alternative in vivo heartworm preventative drug screen, we assessed lymphopenic mouse strains with ablation of the interleukin-2/7 common gamma chain (γc) as susceptible to the larval development phase of Dirofilaria immitis.

RESULTS: Non-obese diabetic (NOD) severe combined immunodeficiency (SCID)γc[-/-] (NSG and NXG) and recombination-activating gene (RAG)2[-/-]γc[-/-] mouse strains yielded viable D. immitis larvae at 2-4 weeks post-infection, including the use of different batches of D. immitis infectious larvae, different D. immitis isolates, and at different laboratories. Mice did not display any clinical signs associated with infection for up to 4 weeks. Developing larvae were found in subcutaneous and muscle fascia tissues, which is the natural site of this stage of heartworm in dogs. Compared with in vitro-propagated larvae at day 14, in vivo-derived larvae had completed the L4 molt, were significantly larger, and contained expanded Wolbachia endobacteria titres. We established an ex vivo L4 paralytic screening system whereby assays with moxidectin or levamisole highlighted discrepancies in relative drug sensitivities in comparison with in vitro-reared L4 D. immitis. We demonstrated effective depletion of Wolbachia by 70%-90% in D. immitis L4 following 2- to 7-day oral in vivo exposures of NSG- or NXG-infected mice with doxycycline or the rapid-acting investigational drug, AWZ1066S. We validated NSG and NXG D. immitis mouse models as a filaricide screen by in vivo treatments with single injections of moxidectin, which mediated a 60%-88% reduction in L4 larvae at 14-28 days.

DISCUSSION: Future adoption of these mouse models will benefit end-user laboratories conducting research and development of novel heartworm preventatives via increased access, rapid turnaround, and reduced costs and may simultaneously decrease the need for experimental cat or dog use.}, } @article {pmid37425681, year = {2023}, author = {Boehm, EC and Jaeger, AS and Ries, HJ and Castañeda, D and Weiler, AM and Valencia, CC and Weger-Lucarelli, J and Ebel, GD and O'Connor, SL and Friedrich, TC and Zamanian, M and Aliota, MT}, title = {Wolbachia -mediated resistance to Zika virus infection in Aedes aegypti is dominated by diverse transcriptional regulation and weak evolutionary pressures.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {37425681}, abstract = {UNLABELLED: A promising candidate for arbovirus control and prevention relies on replacing arbovirus-susceptible Aedes aegypti populations with mosquitoes that have been colonized by the intracellular bacterium Wolbachia and thus have a reduced capacity to transmit arboviruses. This reduced capacity to transmit arboviruses is mediated through a phenomenon referred to as pathogen blocking. Pathogen blocking has primarily been proposed as a tool to control dengue virus (DENV) transmission, however it works against a range of viruses, including Zika virus (ZIKV). Despite years of research, the molecular mechanisms underlying pathogen blocking still need to be better understood. Here, we used RNA-seq to characterize mosquito gene transcription dynamics in Ae. aegypti infected with the w Mel strain of Wolbachia that are being released by the World Mosquito Program in Medellín, Colombia. Comparative analyses using ZIKV-infected, uninfected tissues, and mosquitoes without Wolbachia revealed that the influence of w Mel on mosquito gene transcription is multifactorial. Importantly, because Wolbachia limits, but does not completely prevent, replication of ZIKV and other viruses in coinfected mosquitoes, there is a possibility that these viruses could evolve resistance to pathogen blocking. Therefore, to understand the influence of Wolbachia on within-host ZIKV evolution, we characterized the genetic diversity of molecularly barcoded ZIKV virus populations replicating in Wolbachia -infected mosquitoes and found that within-host ZIKV evolution was subject to weak purifying selection and, unexpectedly, loose anatomical bottlenecks in the presence and absence of Wolbachia . Together, these findings suggest that there is no clear transcriptional profile associated with Wolbachia -mediated ZIKV restriction, and that there is no evidence for ZIKV escape from this restriction in our system.

AUTHOR SUMMARY: When Wolbachia bacteria infect Aedes aegypti mosquitoes, they dramatically reduce the mosquitoes' susceptibility to infection with a range of arthropod-borne viruses, including Zika virus (ZIKV). Although this pathogen-blocking effect has been widely recognized, its mechanisms remain unclear. Furthermore, because Wolbachia limits, but does not completely prevent, replication of ZIKV and other viruses in coinfected mosquitoes, there is a possibility that these viruses could evolve resistance to Wolbachia -mediated blocking. Here, we use host transcriptomics and viral genome sequencing to examine the mechanisms of ZIKV pathogen blocking by Wolbachia and viral evolutionary dynamics in Ae. aegypti mosquitoes. We find complex transcriptome patterns that do not suggest a single clear mechanism for pathogen blocking. We also find no evidence that Wolbachia exerts detectable selective pressures on ZIKV in coinfected mosquitoes. Together our data suggest that it may be difficult for ZIKV to evolve Wolbachia resistance, perhaps due to the complexity of the pathogen blockade mechanism.}, } @article {pmid37424778, year = {2023}, author = {Zhou, JC and Dong, QJ and Shang, D and Ning, SF and Zhang, HH and Wang, Y and Che, WN and Dong, H and Zhang, LS}, title = {Posterior concentration of Wolbachia during the early embryogenesis of the host dynamically shapes the tissue tropism of Wolbachia in host Trichogramma wasps.}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1198428}, pmid = {37424778}, issn = {2235-2988}, mesh = {Animals ; Female ; *Wasps/microbiology ; *Wolbachia/genetics ; In Situ Hybridization, Fluorescence ; Embryonic Development ; Parthenogenesis ; }, abstract = {INTRODUCTION: The bacterial endosymbiont, Wolbachia spp. induce thelytokous parthenogenesis in certain parasitoid wasps, such as the egg parasitoid wasps Trichogramma spp. To complete the cycle of vertical transmission, Wolbachia displays efficient transovarial transmission by targeting the reproductive tissues and often exhibits strong tissue-specific tropism in their host.

METHOD: The present study aimed to describe the basic Wolbachia distribution patterns that occur during the development of Wolbachia-infected, thelytokous Trichogramma dendrolimi, and T. pretiosum. We used fluorescence in situ hybridization (FISH) to investigate Wolbachia signal dynamics during early embryogenesis (from 30 to 120 min). Wolbachia titers and distributions from the embryo to adult stages of Trichogramma after early embryogenesis were detected by absolute quantitative polymerase chain reaction (AQ-PCR) and FISH. The symmetry ratios (SR) of the Wolbachia signals were calculated using the SR odds ratios in the anterior and posterior parts of the host. The SR was determined to describe Wolbachia tropism during early embryogenesis and various developmental stages of Trichogramma.

RESULTS: Wolbachia was concentrated in the posterior part of the embryo during early embryogenesis and the various developmental stages of both T. dendrolimi and T. pretiosum. Wolbachia density increased with the number of nuclei and the initial mitotic division frequency during early embryogenesis. The total Wolbachia titer increased with postembryogenesis development in both T. dendrolimi and T. pretiosum. However, the Wolbachia densities relative to body size were significantly lower at the adult and pupal stages than they were at the embryonic stage.

DISCUSSION: The present work revealed that posterior Wolbachia concentration during early host embryogenesis determined Wolbachia localization in adult wasps. By this mechanism, Wolbachia exhibits efficient vertical transmission across generations by depositing only female Wolbachia-infected offspring. The results of this study describe the dynamics of Wolbachia during the development of their Trichogramma host. The findings of this investigation helped clarify Wolbachia tropism in Trichogramma wasps.}, } @article {pmid37417698, year = {2023}, author = {Yang, J and Chen, Z and Tan, Y and Liu, Z and Cheke, RA}, title = {Threshold dynamics of a stochastic mathematical model for Wolbachia infections.}, journal = {Journal of biological dynamics}, volume = {17}, number = {1}, pages = {2231967}, doi = {10.1080/17513758.2023.2231967}, pmid = {37417698}, issn = {1751-3766}, mesh = {Animals ; *Aedes/microbiology ; *Wolbachia ; Models, Biological ; *Dengue/epidemiology ; Models, Theoretical ; }, abstract = {A stochastic mathematical model is proposed to study how environmental heterogeneity and the augmentation of mosquitoes with Wolbachia bacteria affect the outcomes of dengue disease. The existence and uniqueness of the positive solutions of the system are studied. Then the V-geometrically ergodicity and stochastic ultimate boundedness are investigated. Further, threshold conditions for successful population replacement are derived and the existence of a unique ergodic steady-state distribution of the system is explored. The results show that the ratio of infected to uninfected mosquitoes has a great influence on population replacement. Moreover, environmental noise plays a significant role in control of dengue fever.}, } @article {pmid37417166, year = {2023}, author = {Tharsan, A and Sivabalakrishnan, K and Arthiyan, S and Eswaramohan, T and Raveendran, S and Ramasamy, R and Surendran, SN}, title = {Wolbachia infection is widespread in brackish and fresh water Aedes albopictus (Diptera: Culicidae) in the coastal Jaffna peninsula of northern Sri Lanka.}, journal = {Journal of vector borne diseases}, volume = {60}, number = {2}, pages = {172-178}, doi = {10.4103/0972-9062.361165}, pmid = {37417166}, issn = {0972-9062}, mesh = {Animals ; *Aedes/physiology ; *Wolbachia/genetics ; Sri Lanka ; Phylogeny ; Mosquito Vectors/physiology ; Fresh Water ; *Dengue/prevention & control ; Membrane Proteins/genetics ; }, abstract = {BACKGROUND & OBJECTIVES: Aedes albopictus and Aedes aegypti are important vectors of dengue and many other arboviral diseases in tropical and sub-tropical locations. Both vectors are tolerant of salinity in the dengue-endemic coastal Jaffna peninsula of northern Sri Lanka. Aedes albopictus pre-imaginal stages are found in field brackish water habitats of up to 14 parts per thousand (ppt, gL[-1]) salt in the Jaffna peninsula. Salinity-tolerance in Aedes is characterized by significant genetic and physiological changes. Infection with the wMel strain of the endosymbiont bacterium Wolbachia pipientis reduces dengue transmission in the field by Ae. aegypti, and the same approach is also being considered for Ae. albopictus. In this context, we investigated natural Wolbachia infections in brackish and fresh water field isolates of Ae. albopictus in the Jaffna district.

METHODS: Aedes albopictus collected as pre-imaginal stages using conventional ovitraps in the Jaffna peninsula and adjacent islands of the Jaffna district were screened by PCR utilizing strain-transcending primers for the presence of Wolbachia. Wolbachia strains were then further identified by PCR using strain-specific primers for the Wolbachia surface protein gene wsp. The Jaffna wsp sequences were compared by phylogenetic analysis with other wsp sequences available in Genbank.

RESULTS: Aedes albopictus were found to be widely infected with the wAlbA and wAlbB strains of Wolbachia in Jaffna. The partial wAlbB wsp surface protein gene sequence in Jaffna Ae. albopictus was identical to a corresponding sequence from South India but different from that in mainland Sri Lanka.

Widespread infection of salinity-tolerant Ae. albopictus with Wolbachia is a factor to be considered when developing Wolbachia-based dengue control in coastal areas like the Jaffna peninsula.}, } @article {pmid37416893, year = {2023}, author = {Wenzel, M and Aquadro, CF}, title = {Wolbachia genetically interacts with the bag of marbles germline stem cell gene in male D. melanogaster.}, journal = {microPublication biology}, volume = {2023}, number = {}, pages = {}, pmid = {37416893}, issn = {2578-9430}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; }, abstract = {The bacterial endosymbiont Wolbachia manipulates reproduction of its arthropod hosts to promote its own maternal vertical transmission. In female D. melanogaster , Wolbachia has been shown to genetically interact with three key reproductive genes (bag of marbles (bam) , Sex-lethal, and mei-P26) , as it rescues the reduced female fertility or fecundity phenotype seen in partial loss-of-function mutants of these genes . Here, we show that Wolbachia also partially rescues male fertility in D. melanogaster carrying a new, largely sterile bam allele when in a bam null genetic background. This finding shows that the molecular mechanism of Wolbachia 's influence on its hosts' reproduction involves interaction with genes in males as well as females, at least in D. melanogaster .}, } @article {pmid37415819, year = {2023}, author = {Dong, L and Li, Y and Yang, C and Gong, J and Zhu, W and Huang, Y and Kong, M and Zhao, L and Wang, F and Lu, S and Pu, J and Yang, J}, title = {Species-level microbiota of ticks and fleas from Marmota himalayana in the Qinghai-Tibet Plateau.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1188155}, pmid = {37415819}, issn = {1664-302X}, abstract = {INTRODUCTION: Ticks and fleas, as blood-sucking arthropods, carry and transmit various zoonotic diseases. In the natural plague foci of China, monitoring of Yersinia pestis has been continuously conducted in Marmota himalayana and other host animals, whereas other pathogens carried by vectors are rarely concerned in the Qinghai-Tibet Plateau.

METHODS: In this study, we investigated the microbiota of ticks and fleas sampling from M. himalayana in the Qinghai-Tibet Plateau, China by metataxonomics combined with metagenomic methods.

RESULTS: By metataxonomic approach based on full-length 16S rDNA amplicon sequencing and operational phylogenetic unit (OPU) analyses, we described the microbiota community of ticks and fleas at the species level, annotated 1,250 OPUs in ticks, including 556 known species and 492 potentially new species, accounting for 48.50% and 41.71% of the total reads in ticks, respectively. A total of 689 OPUs were detected in fleas, consisting of 277 known species (40.62% of the total reads in fleas) and 294 potentially new species (56.88%). At the dominant species categories, we detected the Anaplasma phagocytophilum (OPU 421) and potentially pathogenic new species of Wolbachia, Ehrlichia, Rickettsia, and Bartonella. Using shotgun sequencing, we obtained 10 metagenomic assembled genomes (MAGs) from vector samples, including a known species (Providencia heimbachae DFT2), and six new species affliated to four known genera, i.e., Wolbachia, Mumia, Bartonella, and Anaplasma. By the phylogenetic analyses based on full-length 16S rRNA genes and core genes, we identified that ticks harbored pathogenic A. phagocytophilum. Moreover, these potentially pathogenic novel species were more closely related to Ehrlichia muris, Ehrlichia muris subsp. eauclairensis, Bartonella rochalimae, and Rickettsia limoniae, respectively. The OPU 422 Ehrlichia sp1 was most related to Ehrlichia muris and Ehrlichia muris subsp. eauclairensis. The OPU 230 Bartonella sp1 and Bartonella spp. (DTF8 and DTF9) was clustered with Bartonella rochalimae. The OPU 427 Rickettsia sp1 was clustered with Rickettsia limoniae.

DISCUSSION: The findings of the study have advanced our understanding of the potential pathogen groups of vectors in marmot (Marmota himalayana) in the Qinghai-Tibet Plateau.}, } @article {pmid37399133, year = {2023}, author = {Beliavskaia, A and Tan, KK and Sinha, A and Husin, NA and Lim, FS and Loong, SK and Bell-Sakyi, L and Carlow, CKS and AbuBakar, S and Darby, AC and Makepeace, BL and Khoo, JJ}, title = {Metagenomics of culture isolates and insect tissue illuminate the evolution of Wolbachia, Rickettsia and Bartonella symbionts in Ctenocephalides spp. fleas.}, journal = {Microbial genomics}, volume = {9}, number = {7}, pages = {}, pmid = {37399133}, issn = {2057-5858}, support = {BB/P024378/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Rickettsia/genetics ; *Bartonella/genetics ; *Siphonaptera/microbiology ; *Ctenocephalides/microbiology ; *Wolbachia/genetics ; Metagenomics ; Insecta ; }, abstract = {While fleas are often perceived simply as a biting nuisance and a cause of allergic dermatitis, they represent important disease vectors worldwide, especially for bacterial zoonoses such as plague (transmitted by rodent fleas) and some of the rickettsioses and bartonelloses. The cosmopolitan cat (Ctenocephalides felis) and dog (Ctenocephalides canis) fleas, as well as Ctenocephalides orientis (restricted to tropical and subtropical Asia), breed in human dwellings and are vectors of cat-scratch fever (caused by Bartonella spp.) and Rickettsia spp., including Rickettsia felis (agent of flea-borne spotted fever) and Rickettsia asembonensis , a suspected pathogen. These Rickettsia spp. are members of a phylogenetic clade known as the ‘transitional group’, which includes both human pathogens and arthropod-specific endosymbionts. The relatively depauperate flea microbiome can also contain other endosymbionts, including a diverse range of Wolbachia strains. Here, we present circularized genome assemblies for two C. orientis-derived pathogens (Bartonella clarridgeiae and R. asembonensis) from Malaysia, a novel Wolbachia strain (wCori), and the C. orientis mitochondrion; all were obtained by direct metagenomic sequencing of flea tissues. Moreover, we isolated two Wolbachia strains from Malaysian C. felis into tick cell culture and recovered circularized genome assemblies for both, one of which (wCfeF) is newly sequenced. We demonstrate that the three Wolbachia strains are representatives of different major clades (‘supergroups’), two of which appear to be flea-specific. These Wolbachia genomes exhibit unique combinations of features associated with reproductive parasitism or mutualism, including prophage WO, cytoplasmic incompatibility factors and the biotin operon of obligate intracellular microbes. The first circularized assembly for R. asembonensis includes a plasmid with a markedly different structure and gene content compared to the published plasmid; moreover, this novel plasmid was also detected in cat flea metagenomes from the USA. Analysis of loci under positive selection in the transitional group revealed genes involved in host–pathogen interactions that may facilitate host switching. Finally, the first B. clarridgeiae genome from Asia exhibited large-scale genome stability compared to isolates from other continents, except for SNPs in regions predicted to mediate interactions with the vertebrate host. These findings highlight the paucity of data on the genomic diversity of Ctenocephalides-associated bacteria and raise questions regarding how interactions between members of the flea microbiome might influence vector competence.}, } @article {pmid37397989, year = {2023}, author = {Pujhari, S and Hughes, GL and Pakpour, N and Suzuki, Y and Rasgon, JL}, title = {Wolbachia-induced inhibition of O'nyong nyong virus in Anopheles mosquitoes is mediated by Toll signaling and modulated by cholesterol.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {37397989}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; }, abstract = {Enhanced host immunity and competition for metabolic resources are two main competing hypotheses for the mechanism of Wolbachia-mediated pathogen inhibition in arthropods. Using an Anopheles mosquito - somatic Wolbachia infection - O'nyong nyong virus (ONNV) model, we demonstrate that the mechanism underpinning Wolbachia-mediated virus inhibition is up-regulation of the Toll innate immune pathway. However, the viral inhibitory properties of Wolbachia were abolished by cholesterol supplementation. This result was due to Wolbachia-dependent cholesterol-mediated suppression of Toll signaling rather than competition for cholesterol between Wolbachia and virus. The inhibitory effect of cholesterol was specific to Wolbachia-infected Anopheles mosquitoes and cells. These data indicate that both Wolbachia and cholesterol influence Toll immune signaling in Anopheles mosquitoes in a complex manner and provide a functional link between the host immunity and metabolic competition hypotheses for explaining Wolbachia-mediated pathogen interference in mosquitoes. In addition, these results provide a mechanistic understanding of the mode of action of Wolbachia-induced pathogen blocking in Anophelines, which is critical to evaluate the long-term efficacy of control strategies for malaria and Anopheles-transmitted arboviruses.}, } @article {pmid37391513, year = {2023}, author = {Abel, SM and Hong, Z and Williams, D and Ireri, S and Brown, MQ and Su, T and Hung, KY and Henke, JA and Barton, JP and Le Roch, KG}, title = {Small RNA sequencing of field Culex mosquitoes identifies patterns of viral infection and the mosquito immune response.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {10598}, pmid = {37391513}, issn = {2045-2322}, support = {U01 CK000516/CK/NCEZID CDC HHS/United States ; S10 OD026929/OD/NIH HHS/United States ; }, mesh = {Humans ; Animals ; *Culex ; *Culicidae ; Mosquito Vectors ; *Virus Diseases ; Antiviral Agents ; }, abstract = {Mosquito-borne disease remains a significant burden on global health. In the United States, the major threat posed by mosquitoes is transmission of arboviruses, including West Nile virus by mosquitoes of the Culex genus. Virus metagenomic analysis of mosquito small RNA using deep sequencing and advanced bioinformatic tools enables the rapid detection of viruses and other infecting organisms, both pathogenic and non-pathogenic to humans, without any precedent knowledge. In this study, we sequenced small RNA samples from over 60 pools of Culex mosquitoes from two major areas of Southern California from 2017 to 2019 to elucidate the virome and immune responses of Culex. Our results demonstrated that small RNAs not only allowed the detection of viruses but also revealed distinct patterns of viral infection based on location, Culex species, and time. We also identified miRNAs that are most likely involved in Culex immune responses to viruses and Wolbachia bacteria, and show the utility of using small RNA to detect antiviral immune pathways including piRNAs against some pathogens. Collectively, these findings show that deep sequencing of small RNA can be used for virus discovery and surveillance. One could also conceive that such work could be accomplished in various locations across the world and over time to better understand patterns of mosquito infection and immune response to many vector-borne diseases in field samples.}, } @article {pmid37381006, year = {2023}, author = {Zeb, J and Song, B and Khan, MA and Senbill, H and Aziz, MU and Hussain, S and Waris, A and E-Tabor, A and Sparagano, OA}, title = {Genetic diversity of vector-borne pathogens in ixodid ticks infesting dogs from Pakistan with notes on Ehrlichia canis, Rickettsia raoultii and Dirofilaria immitis detection.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {214}, pmid = {37381006}, issn = {1756-3305}, support = {9380108//Olivier Andre Sparagano/ ; }, mesh = {Humans ; Dogs ; Animals ; Ehrlichia canis/genetics ; *Dirofilaria immitis ; Pakistan/epidemiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Ixodidae ; *Rickettsia/genetics ; Ehrlichia/genetics ; *Canidae ; Dirofilaria ; Genetic Variation ; }, abstract = {BACKGROUND: Vector-/tick-borne pathogens (V/TBPs) pose a potential threat to human and animal health globally. Information regarding canine V/TBPs is scarce and no specific study has been conducted so far to explore the microbial diversity within ticks infesting dogs from Pakistan. Herein, this knowledge gap is addressed by assessing the genetic diversity and prevalence pattern of V/TBPs in ixodid ticks with special implications for public and canine health.

METHODS: A total of 1150 hard ticks were collected from 300 dogs across central Khyber Pakhtunkhwa (KP), Pakistan. After morpho-molecular identification, 120 tick samples were screened for the presence of V/TBPs by amplifying 16S rRNA/gltA (Rickettsia/Ehrlichia and Wolbachia sp.), 18S rRNA (Theileria sp.) and cox1 (Dirofilaria sp.) genes through PCR followed by sequencing and phylogenetic study.

RESULTS: In toto, 50 ixodid ticks (50/120, 41.7%) were found positive for V/TBPs DNA. The detected V/TBPs were categorized into five genera and eight species, viz. Ehrlichia (E. canis and Ehrlichia sp.), Rickettsia (R. massiliae, R. raoultii and Rickettsia sp.), Theileria (T. annulata), Dirofilaria (D. immitis) and Wolbachia (Wolbachia sp.). The pathogen prevalence patterns showed that R. massiliae was the most prevalent zoonotic V/TBP (19.5%), followed by E. canis (10.8%), Rickettsia sp. (7.5%), R. raoultii (6.7%), T. annulata (5.8%), D. immitis (5.8%), Wolbachia sp. (4.2%) and Ehrlichia sp. (3.3%), respectively. Among the screened tick species, most Rhipicephalus sanguineus sensu lato samples were found positive for V/TBP DNA (20/20,100%) followed by Rh. turanicus sensu stricto (13/20, 65%), Hyalomma dromedarii (8/20, 40%), Rh. haemaphysaloides (6/20, 30%), Hy. excavatum (2/20, 10%) and Rh. microplus (1/20, 5%). Co-occurrence of V/TBP was also detected in tick specimens (single V/TBP infection: 32 ticks; double and triple: 13 and 5 tick samples). The detected pathogens shared a phylogenetic relationship with similar isolates published in NCBI GenBank from Old and New World countries.

CONCLUSION: Ixodid ticks infesting dogs harbor a diverse array of V/TBPs including zoonotic agents from Pakistan. Furthermore, the presence of D. immitis in ticks that infest dogs raises the possibility that this parasite has either attained its dead-end host (i.e. the tick) while feeding on dogs or has expanded its range of intermediate/paratenic hosts. Further research work is needed to investigate the epidemiology and confirm the vector competence of screened tick species for these pathogens from Pakistan.}, } @article {pmid37376609, year = {2023}, author = {Baltar, JMC and Pavan, MG and Corrêa-Antônio, J and Couto-Lima, D and Maciel-de-Freitas, R and David, MR}, title = {Gut Bacterial Diversity of Field and Laboratory-Reared Aedes albopictus Populations of Rio de Janeiro, Brazil.}, journal = {Viruses}, volume = {15}, number = {6}, pages = {}, pmid = {37376609}, issn = {1999-4915}, mesh = {Animals ; Female ; *Aedes ; *Zika Virus/genetics ; *Gastrointestinal Microbiome ; *Zika Virus Infection ; Brazil ; RNA, Ribosomal, 16S/genetics ; Mosquito Vectors ; Bacteria/genetics ; }, abstract = {BACKGROUND: The mosquito microbiota impacts different parameters in host biology, such as development, metabolism, immune response and vector competence to pathogens. As the environment is an important source of acquisition of host associate microbes, we described the microbiota and the vector competence to Zika virus (ZIKV) of Aedes albopictus from three areas with distinct landscapes.

METHODS: Adult females were collected during two different seasons, while eggs were used to rear F1 colonies. Midgut bacterial communities were described in field and F1 mosquitoes as well as in insects from a laboratory colony (>30 generations, LAB) using 16S rRNA gene sequencing. F1 mosquitoes were infected with ZIKV to determine virus infection rates (IRs) and dissemination rates (DRs). Collection season significantly affected the bacterial microbiota diversity and composition, e.g., diversity levels decreased from the wet to the dry season. Field-collected and LAB mosquitoes' microbiota had similar diversity levels, which were higher compared to F1 mosquitoes. However, the gut microbiota composition of field mosquitoes was distinct from that of laboratory-reared mosquitoes (LAB and F1), regardless of the collection season and location. A possible negative correlation was detected between Acetobacteraceae and Wolbachia, with the former dominating the gut microbiota of F1 Ae. albopictus, while the latter was absent/undetectable. Furthermore, we detected significant differences in infection and dissemination rates (but not in the viral load) between the mosquito populations, but it does not seem to be related to gut microbiota composition, as it was similar between F1 mosquitoes regardless of their population.

CONCLUSIONS: Our results indicate that the environment and the collection season play a significant role in shaping mosquitoes' bacterial microbiota.}, } @article {pmid37375183, year = {2023}, author = {Han, MJ and Pan, M and Xiao, G and Yuan, Y and Chen, S and Zou, Z}, title = {Assessing Boron-Pleuromutilin AN11251 for the Development of Antibacterial Agents.}, journal = {Molecules (Basel, Switzerland)}, volume = {28}, number = {12}, pages = {}, pmid = {37375183}, issn = {1420-3049}, support = {0/GATES/Bill & Melinda Gates Foundation/United States ; }, mesh = {Humans ; Anti-Bacterial Agents/chemistry ; Boron ; *Diterpenes/chemistry ; *Polycyclic Compounds/pharmacology ; *Onchocerciasis/drug therapy ; Bacteria ; Gram-Positive Bacteria ; Microbial Sensitivity Tests ; Pleuromutilins ; }, abstract = {Pleuromutilins are a group of antibiotics derived from the naturally occurring compound. The recent approval of lefamulin for both intravenous and oral doses in humans to treat community-acquired bacterial pneumonia has prompted investigations in modifying the structure to broaden the antibacterial spectrum, enhance the activity, and improve the pharmacokinetic properties. AN11251 is a C(14)-functionalized pleuromutilin with a boron-containing heterocycle substructure. It was demonstrated to be an anti-Wolbachia agent with therapeutic potential for Onchocerciasis and lymphatic filariasis. Here, the in vitro and in vivo PK parameters of AN11251 were measured including PPB, intrinsic clearance, half-life, systemic clearance, and volume of distribution. The results indicate that the benzoxaborole-modified pleuromutilin possesses good ADME and PK properties. AN11251 has potent activities against the Gram-positive bacterial pathogens tested, including various drug-resistant strains, and against the slow-growing mycobacterial species. Finally, we employed PK/PD modeling to predict the human dose for treatment of disease caused by Wolbachia, Gram-positive bacteria, or Mycobacterium tuberculosis, which might facilitate the further development of AN11251.}, } @article {pmid37370177, year = {2023}, author = {Angstmann, H and Pfeiffer, S and Kublik, S and Ehrhardt, B and Uliczka, K and Rabe, KF and Roeder, T and Wagner, C and Schloter, M and Krauss-Etschmann, S}, title = {The microbial composition of larval airways from Drosophila melanogaster differ between specimens from laboratory and natural habitats.}, journal = {Environmental microbiome}, volume = {18}, number = {1}, pages = {55}, pmid = {37370177}, issn = {2524-6372}, abstract = {BACKGROUND: The fruit fly Drosophila melanogaster lives in natural habitats and has also long been used as a model organism in biological research. In this study, we used a molecular barcoding approach to analyse the airways microbiome of larvae of D. melanogaster, which were obtained from eggs of flies of the laboratory strain w[1118] and from immune deficient flies (NF-kB-K), and from wild-caught flies. To assess intergenerational transmission of microbes, all eggs were incubated under the same semi-sterile conditions.

RESULTS: The airway microbiome of larvae from both lab-strains was dominated by the two families Acetobacteraceae and Lactobacillaceae, while larvae from wild-caught flies were dominated by Lactobacillaceae, Anaplasmataceae and Leuconostocaceae. Barcodes linked to Anaplasmataceae could be further assigned to Wolbachia sp., which is a widespread intracellular pathogen in arthropods. For Leuconostoceae, the most abundant reads were assigned to Weissella sp. Both Wolbachia and Weissella affect the development of the insects. Finally, a relative high abundance of Serratia sp. was found in larvae from immune deficient relish[-/-] compared to w[1118] and wild-caught fly airways.

CONCLUSIONS: Our results show for the first time that larvae from D. melanogaster harbor an airway microbiome, which is of low complexity and strongly influenced by the environmental conditions and to a lesser extent by the immune status. Furthermore, our data indicate an intergenerational transmission of the microbiome as shaped by the environment.}, } @article {pmid37367374, year = {2023}, author = {Hu, D and Li, W and Wang, J and Peng, Y and Yun, Y and Peng, Y}, title = {Interaction of High Temperature Stress and Wolbachia Infection on the Biological Characteristic of Drosophila melanogaster.}, journal = {Insects}, volume = {14}, number = {6}, pages = {}, pmid = {37367374}, issn = {2075-4450}, support = {2019020701011464//Wuhan Science and Technology Bureau/ ; 2018FY100400//National Science and Technology Basic Research Program of China/ ; }, abstract = {It was reported that temperature affects the distribution of Wolbachia in the host, but only a few papers reported the effect of the interaction between high temperature and Wolbachia on the biological characteristic of the host. Here, we set four treatment Drosophila melanogaster groups: Wolbachia-infected flies in 25 °C (W[+]M), Wolbachia-infected flies in 31 °C (W[+]H), Wolbachia-uninfected flies in 25 °C (W[-]M), Wolbachia-uninfected flies in 31 °C (W[-]H), and detected the interaction effect of temperature and Wolbachia infection on the biological characteristic of D. melanogaster in F1, F2 and F3 generations. We found that both temperature and Wolbachia infection had significant effects on the development and survival rate of D. melanogaster. High temperature and Wolbachia infection had interaction effect on hatching rate, developmental durations, emergence rate, body weight and body length of F1, F2 and F3 flies, and the interaction effect also existed on oviposition amount of F3 flies, and on pupation rate of F2 and F3 flies. High temperature stress reduced the Wolbachia vertical transmission efficiency between generations. These results indicated that high temperature stress and Wolbachia infection had negative effects on the morphological development of D. melanogaster.}, } @article {pmid37367332, year = {2023}, author = {Fallon, AM and Carroll, EM}, title = {Virus-like Particles from Wolbachia-Infected Cells May Include a Gene Transfer Agent.}, journal = {Insects}, volume = {14}, number = {6}, pages = {}, pmid = {37367332}, issn = {2075-4450}, abstract = {Wolbachia are obligate intracellular bacteria that occur in insects and filarial worms. Strains that infect insects have genomes that encode mobile genetic elements, including diverse lambda-like prophages called Phage WO. Phage WO packages an approximately 65 kb viral genome that includes a unique eukaryotic association module, or EAM, that encodes unusually large proteins thought to mediate interactions between the bacterium, its virus, and the eukaryotic host cell. The Wolbachia supergroup B strain, wStri from the planthopper Laodelphax striatellus, produces phage-like particles that can be recovered from persistently infected mosquito cells by ultracentrifugation. Illumina sequencing, assembly, and manual curation of DNA from two independent preparations converged on an identical 15,638 bp sequence that encoded packaging, assembly, and structural proteins. The absence of an EAM and regulatory genes defined for Phage WO from the wasp, Nasonia vitripennis, was consistent with the possibility that the 15,638 bp sequence represents an element related to a gene transfer agent (GTA), characterized by a signature head-tail region encoding structural proteins that package host chromosomal DNA. Future investigation of GTA function will be supported by the improved recovery of physical particles, electron microscopic examination of potential diversity among particles, and rigorous examination of DNA content by methods independent of sequence assembly.}, } @article {pmid37363264, year = {2023}, author = {Gashururu, RS and Maingi, N and Githigia, SM and Getange, DO and Ntivuguruzwa, JB and Habimana, R and Cecchi, G and Gashumba, J and Bargul, JL and Masiga, DK}, title = {Trypanosomes infection, endosymbionts, and host preferences in tsetse flies (Glossina spp.) collected from Akagera park region, Rwanda: A correlational xenomonitoring study.}, journal = {One health (Amsterdam, Netherlands)}, volume = {16}, number = {}, pages = {100550}, pmid = {37363264}, issn = {2352-7714}, abstract = {Akagera National Park and its surroundings are home to tsetse flies and a number of their mammalian hosts in Rwanda. A One-health approach is being used in the control and surveillance of both animal and human trypanosomosis in Rwanda. Determination of the infection level in tsetse flies, species of trypanosomes circulating in vectors, the source of tsetse blood meal and endosymbionts is crucial in understanding the epidemiology of the disease in animals and humans in the region. Tsetse flies (n = 1101), comprising Glossina pallidipes (n = 771) and Glossina morsitans centralis (n = 330) were collected from Akagera park and surrounding areas between May 2018 and June 2019. The flies were screened for trypanosomes, vertebrate host DNA to identify sources of blood meal, and endosymbionts by PCR - High Resolution Melting analysis and amplicon sequencing. The feeding frequency and the feeding indices (selection index - W) were calculated to identify the preferred hosts. An overall trypanosome infection rate of 13.9% in the fly's Head and Proboscis (HP) and 24.3% in the Thorax and Abdomen (TA) were found. Eight trypanosome species were identified in the tsetse fly HP and TA, namely: Trypanosoma (T.) brucei brucei, T. congolense Kilifi, T. congolense savannah, T. vivax, T. simiae, T. evansi, T. godfreyi, T. grayi and T. theileri. We found no evidence of human-infective T. brucei rhodesiense. We also identified eighteen species of vertebrate hosts that tsetse flies fed on, and the most frequent one was the buffalo (Syncerus caffer) (36.5%). The frequently detected host by selection index was the rhinoceros (Diceros bicornis) (W = 16.2). Most trypanosome infections in tsetse flies were associated with the buffalo blood meal. The prevalence of tsetse endosymbionts Sodalis and Wolbachia was 2.8% and 4.8%, respectively. No Spiroplasma and Salivary Gland Hypertrophy Virus were detected. These findings implicate the buffaloes as the important reservoirs of tsetse-transmitted trypanosomes in the area. This contributes to predicting the main cryptic reservoirs and therefore guiding the effective control of the disease. The study findings provide the key scientific information that supports the current One Health collaboration in the control and surveillance of tsetse-transmitted trypanosomosis in Rwanda.}, } @article {pmid37362913, year = {2023}, author = {Beckmann, J and Gillespie, J and Tauritz, D}, title = {Modeling emergence of Wolbachia toxin-antidote protein functions with an evolutionary algorithm.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1116766}, pmid = {37362913}, issn = {1664-302X}, abstract = {Evolutionary algorithms (EAs) simulate Darwinian evolution and adeptly mimic natural evolution. Most EA applications in biology encode high levels of abstraction in top-down population ecology models. In contrast, our research merges protein alignment algorithms from bioinformatics into codon based EAs that simulate molecular protein string evolution from the bottom up. We apply our EA to reconcile a problem in the field of Wolbachia induced cytoplasmic incompatibility (CI). Wolbachia is a microbial endosymbiont that lives inside insect cells. CI is conditional insect sterility that operates as a toxin antidote (TA) system. Although, CI exhibits complex phenotypes not fully explained under a single discrete model. We instantiate in-silico genes that control CI, CI factors (cifs), as strings within the EA chromosome. We monitor the evolution of their enzymatic activity, binding, and cellular localization by applying selective pressure on their primary amino acid strings. Our model helps rationalize why two distinct mechanisms of CI induction might coexist in nature. We find that nuclear localization signals (NLS) and Type IV secretion system signals (T4SS) are of low complexity and evolve fast, whereas binding interactions have intermediate complexity, and enzymatic activity is the most complex. Our model predicts that as ancestral TA systems evolve into eukaryotic CI systems, the placement of NLS or T4SS signals can stochastically vary, imparting effects that might impact CI induction mechanics. Our model highlights how preconditions and sequence length can bias evolution of cifs toward one mechanism or another.}, } @article {pmid37360524, year = {2023}, author = {Ratnayake, OC and Chotiwan, N and Saavedra-Rodriguez, K and Perera, R}, title = {The buzz in the field: the interaction between viruses, mosquitoes, and metabolism.}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1128577}, pmid = {37360524}, issn = {2235-2988}, support = {R01 AI121211/AI/NIAID NIH HHS/United States ; R01 AI151166/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Humans ; *Dengue Virus/physiology ; *Aedes ; *Arboviruses ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {Among many medically important pathogens, arboviruses like dengue, Zika and chikungunya cause severe health and economic burdens especially in developing countries. These viruses are primarily vectored by mosquitoes. Having surmounted geographical barriers and threat of control strategies, these vectors continue to conquer many areas of the globe exposing more than half of the world's population to these viruses. Unfortunately, no medical interventions have been capable so far to produce successful vaccines or antivirals against many of these viruses. Thus, vector control remains the fundamental strategy to prevent disease transmission. The long-established understanding regarding the replication of these viruses is that they reshape both human and mosquito host cellular membranes upon infection for their replicative benefit. This leads to or is a result of significant alterations in lipid metabolism. Metabolism involves complex chemical reactions in the body that are essential for general physiological functions and survival of an organism. Finely tuned metabolic homeostases are maintained in healthy organisms. However, a simple stimulus like a viral infection can alter this homeostatic landscape driving considerable phenotypic change. Better comprehension of these mechanisms can serve as innovative control strategies against these vectors and viruses. Here, we review the metabolic basis of fundamental mosquito biology and virus-vector interactions. The cited work provides compelling evidence that targeting metabolism can be a paradigm shift and provide potent tools for vector control as well as tools to answer many unresolved questions and gaps in the field of arbovirology.}, } @article {pmid37349802, year = {2023}, author = {Benkacimi, L and Diarra, AZ and Bompar, JM and Bérenger, JM and Parola, P}, title = {Microorganisms associated with hedgehog arthropods.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {211}, pmid = {37349802}, issn = {1756-3305}, mesh = {Animals ; Humans ; *Arthropods/microbiology ; Hedgehogs/parasitology ; *Rickettsia ; *Bartonella ; Mammals ; *Siphonaptera/microbiology ; *Ticks/microbiology ; *Flea Infestations ; }, abstract = {Hedgehogs are small synanthropic mammals that live in rural areas as well as in urban and suburban areas. They can be reservoirs of several microorganisms, including certain pathogenic agents that cause human and animal public health issues. Hedgehogs are often parasitized by blood-sucking arthropods, mainly hard ticks and fleas, which in turn can also carry various vector-born microorganisms of zoonotic importance. Many biotic factors, such as urbanization and agricultural mechanization, have resulted in the destruction of the hedgehog's natural habitats, leading these animals to take refuge near human dwellings, seeking food and shelter in parks and gardens and exposing humans to zoonotic agents that can be transmitted either directly by them or indirectly by their ectoparasites. In this review, we focus on the microorganisms detected in arthropods sampled from hedgehogs worldwide. Several microorganisms have been reported in ticks collected from these animals, including various Borrelia spp., Anaplasma spp., Ehrlichia spp., and Rickettsia spp. species as well as Coxiella burnetii and Leptospira spp. As for fleas, C. burnetii, Rickettsia spp., Wolbachia spp., Mycobacterium spp. and various Bartonella species have been reported. The detection of these microorganisms in arthropods does not necessarily mean that they can be transmitted to humans and animals. While the vector capacity and competence of fleas and ticks for some of these microorganisms has been proven, in other cases the microorganisms may have simply been ingested with blood taken from an infected host. Further investigations are needed to clarify this issue. As hedgehogs are protected animals, handling them is highly regulated, making it difficult to conduct epidemiological studies on them. Their ectoparasites represent a very interesting source of information on microorganisms circulating in populations of these animals, especially vector-born ones.}, } @article {pmid37347767, year = {2023}, author = {Ho, SH and Lim, JT and Ong, J and Hapuarachchi, HC and Sim, S and Ng, LC}, title = {Singapore's 5 decades of dengue prevention and control-Implications for global dengue control.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {6}, pages = {e0011400}, pmid = {37347767}, issn = {1935-2735}, mesh = {Aged ; Animals ; Humans ; Seroepidemiologic Studies ; Singapore/epidemiology ; *Aedes ; Climate Change ; *Dengue/epidemiology/prevention & control ; }, abstract = {This paper summarises the lessons learnt in dengue epidemiology, risk factors, and prevention in Singapore over the last half a century, during which Singapore evolved from a city of 1.9 million people to a highly urban globalised city-state with a population of 5.6 million. Set in a tropical climate, urbanisation among green foliage has created ideal conditions for the proliferation of Aedes aegypti and Aedes albopictus, the mosquito vectors that transmit dengue. A vector control programme, largely for malaria, was initiated as early as 1921, but it was only in 1966 that the Vector Control Unit (VCU) was established to additionally tackle dengue haemorrhagic fever (DHF) that was first documented in the 1960s. Centred on source reduction and public education, and based on research into the bionomics and ecology of the vectors, the programme successfully reduced the Aedes House Index (HI) from 48% in 1966 to <5% in the 1970s. Further enhancement of the programme, including through legislation, suppressed the Aedes HI to around 1% from the 1990s. The current programme is characterised by 4 key features: (i) proactive inter-epidemic surveillance and control that is stepped up during outbreaks; (ii) risk-based prevention and intervention strategies based on advanced data analytics; (iii) coordinated inter-sectoral cooperation between the public, private, and people sectors; and (iv) evidence-based adoption of new tools and strategies. Dengue seroprevalence and force of infection (FOI) among residents have substantially and continuously declined over the 5 decades. This is consistent with the observation that dengue incidence has been delayed to adulthood, with severity highest among the elderly. Paradoxically, the number of reported dengue cases and outbreaks has increased since the 1990s with record-breaking epidemics. We propose that Singapore's increased vulnerability to outbreaks is due to low levels of immunity in the population, constant introduction of new viral variants, expanding urban centres, and increasing human density. The growing magnitude of reported outbreaks could also be attributed to improved diagnostics and surveillance, which at least partially explains the discord between rising trend in cases and the continuous reduction in dengue seroprevalence. Changing global and local landscapes, including climate change, increasing urbanisation and global physical connectivity are expected to make dengue control even more challenging. The adoption of new vector surveillance and control tools, such as the Gravitrap and Wolbachia technology, is important to impede the growing threat of dengue and other Aedes-borne diseases.}, } @article {pmid37347285, year = {2023}, author = {Ehlers, LP and Slaviero, M and De Lorenzo, C and Fagundes-Moreira, R and de Souza, VK and Perles, L and Baggio-Souza, V and Bezerra-Santos, MA and Modrý, D and Benovics, M and Panziera, W and Driemeier, D and Pavarini, SP and Soares, JF and Otranto, D and Sonne, L}, title = {Pathological findings associated with Dipetalonema spp. (Spirurida, Onchocercidae) infection in two species of Neotropical monkeys from Brazil.}, journal = {Parasitology research}, volume = {122}, number = {9}, pages = {1973-1982}, pmid = {37347285}, issn = {1432-1955}, mesh = {Animals ; *Dipetalonema/genetics ; *Spirurida/genetics ; Brazil/epidemiology ; Haplorhini/genetics ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Filarioidea/genetics ; *Dipetalonema Infections/parasitology ; *Nematoda/genetics ; }, abstract = {Among vector-borne helminths, filarioids of the genus Dipetalonema (Spirurida: Onchocercidae) localize in several tissues and body cavities of several animal species, causing mild to moderate lesions. The pathological findings associated with Dipetalonema spp. infection in Neotropical monkeys from southern Brazil are herein described, along with a fatal case due to filarial polyserositis and entrapment of an intestinal segment. At necropsy, nematodes were observed in abdominal and thoracic cavities, or in the pericardium of 37 (31.3%) out of the 118 individuals examined (i.e., 35 Alouatta guariba clamitans and two Sapajus nigritus). In addition, at histology, 27.0% of positive animals presented microfilarie (inside blood vessels of lung, spleen, liver, and brain) and 8.1% presented adult nematodes in the heart, lung, and liver. In two cases, cross-sections of filarioids were associated with areas of epicardial thickening with intense fibrosis and pyogranulomatous inflammation in the brain, heart, liver, lungs, or spleen. The DNA fragment was amplify using the cox1 gene, sequenced and analyzed to identify the nematode species collected; presence of Wolbachia was assessed in the filarioids using the 16S rRNA gene. At BLAST analysis of the cox1 gene, 10 sequences showed 91.7% nucleotide identity with Dipetalonema gracile, and two with D. gracile (98.5%) and Dipetalonema graciliformis (98.3%). Phylogenetic analyses clustered sequences of the cox1 obtained in this study in two clades corresponding with the host species. Wolbachia sp. endosymbiont was detected in four samples. Data herein reported provide a description of pathological lesions associated with the infection by Dipetalonema spp., suggesting that they may cause disease in Neotropical monkeys. In addition, a better understanding of diversity and biology of Dipetalonema spp. in South America is needed to assess the impact they may cause in native non-human primates from Brazil.}, } @article {pmid37324163, year = {2023}, author = {Zhu, X and Zhang, L and Li, J and He, A and You, M and You, S}, title = {Effects of Antibiotic Treatment on the Development and Bacterial Community of the Wolbachia-Infected Diamondback Moth.}, journal = {Evolutionary bioinformatics online}, volume = {19}, number = {}, pages = {11769343231175269}, pmid = {37324163}, issn = {1176-9343}, abstract = {Based on the important role of antibiotic treatment in the research of the interaction between Wolbachia and insect hosts, this study aimed to identify the most suitable antibiotic and concentration for Wolbachia elimination in the P. xylostella, and to investigate the effect of Wolbachia and antibiotic treatment on the bacterial community of P. xylostella. Our results showed that the Wolbachia-infected strain was plutWB1 of supergroup B in the P. xylostella population collected in Nepal in this study; 1 mg/mL rifampicin could remove Wolbachia infection in P. xylostella after 1 generation of feeding treatment and the toxic effect was relatively low; among the 29 samples of adult P. xylostella in our study (10 WU samples, 10 WA samples, and 9 WI samples), 52.5% of the sequences were of Firmicutes and 47.5% were of Proteobacteria, with the dominant genera being mainly Carnobacterium (46.2%), Enterobacter (10.1%), and Enterococcus (6.2%); Moreover, antibiotic removal of Wolbachia infection in P. xylostella and transfer to normal conditions for 10 generations no longer significantly affected the bacterial community of P. xylostella. This study provides a theoretical basis for the elimination method of Wolbachia in the P. xylostella, as well as a reference for the elimination method of Wolbachia in other Wolbachia-infected insect species, and a basis for the study of the extent and duration of the effect of antibiotic treatment on the bacterial community of the P. xylostella.}, } @article {pmid37318795, year = {2023}, author = {Zhang, DJ and Sun, Y and Yamada, H and Wu, Y and Wang, G and Feng, QD and Paerhande, D and Maiga, H and Bouyer, J and Qian, J and Wu, ZD and Zheng, XY}, title = {Effects of radiation on the fitness, sterility and arbovirus susceptibility of a Wolbachia-free Aedes albopictus strain for use in the sterile insect technique.}, journal = {Pest management science}, volume = {79}, number = {11}, pages = {4186-4196}, doi = {10.1002/ps.7615}, pmid = {37318795}, issn = {1526-4998}, support = {//Guangzhou Basic and Applied Basic Research Foundation/ ; //IAEA Coordinated Research Project/ ; //IAEA Department of Technical Cooperation/ ; //National Key Research and Development Program of China/ ; //National Natural Science Foundation of China/ ; //NSFC-BMGF/ ; //Nuclear Energy R & D Project/ ; //Open Foundation of NHC Key Laboratory of Tropical Disease Control, Hainan Medical University/ ; //Science and Technology Planning Project of Guangdong Province, China/ ; }, abstract = {BACKGROUND: The sterile insect technique (SIT) is a green and species-specific insect pest control technique that suppresses target populations by releasing factory-reared, radiosterilized males into the wild. Once released, it is important to be able to distinguish the released males from the wild males for monitoring purposes. Several methods to mark the sterile males exist. However, most have limitations due to monetary, process efficiency, or insect quality. Aedes albopictus is naturally infected with Wolbachia at a high prevalence, therefore the elimination of Wolbachia can serve as a biomarker to distinguish factory-reared male mosquitoes from wild conspecifics.

RESULTS: In this study, a Wolbachia-free Ae. albopictus GT strain was developed and its fitness evaluated, which was found to be comparable to the wild GUA strain. In addition, GT male mosquitoes were irradiated at the adult stage and a dose of 20 Gy or more induced over 99% sterility. Moreover, a dose of 30 Gy (almost completely sterilizing male and female mosquitoes) had limited effects on the mating competitiveness of GT males and the vector competence of GT females, respectively. However, radiation reduced mosquito longevity, regardless of sex.

CONCLUSION: Our results indicate that the Ae. albopictus GT strain can be distinguished from wild mosquitoes based on Wolbachia status and shows similar fitness, radio-sensitivity and arbovirus susceptibility to the GUA strain, indicating that it is feasible to use the GT strain to suppress Ae. albopictus populations for SIT programmes. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.}, } @article {pmid37317119, year = {2023}, author = {Santos, NACD and Carvalho, VR and Souza-Neto, JA and Alonso, DP and Ribolla, PEM and Medeiros, JF and Araujo, MDS}, title = {Bacterial Microbiota from Lab-Reared and Field-Captured Anopheles darlingi Midgut and Salivary Gland.}, journal = {Microorganisms}, volume = {11}, number = {5}, pages = {}, pmid = {37317119}, issn = {2076-2607}, support = {INV-003970/GATES/Bill & Melinda Gates Foundation/United States ; }, abstract = {Anopheles darlingi is a major malaria vector in the Amazon region and, like other vectors, harbors a community of microorganisms with which it shares a network of interactions. Here, we describe the diversity and bacterial composition from the midguts and salivary glands of lab-reared and field-captured An. darlingi using metagenome sequencing of the 16S rRNA gene. The libraries were built using the amplification of the region V3-V4 16S rRNA gene. The bacterial community from the salivary glands was more diverse and richer than the community from the midguts. However, the salivary glands and midguts only showed dissimilarities in beta diversity between lab-reared mosquitoes. Despite that, intra-variability was observed in the samples. Acinetobacter and Pseudomonas were dominant in the tissues of lab-reared mosquitoes. Sequences of Wolbachia and Asaia were both found in the tissue of lab-reared mosquitoes; however, only Asaia was found in field-captured An. darlingi, but in low abundance. This is the first report on the characterization of microbiota composition from the salivary glands of An. darlingi from lab-reared and field-captured individuals. This study can provide invaluable insights for future investigations regarding mosquito development and interaction between mosquito microbiota and Plasmodium sp.}, } @article {pmid37314477, year = {2023}, author = {An, Y and Braga, MP and Garcia, SL and Grudzinska-Sterno, M and Hambäck, PA}, title = {Host Phylogeny Structures the Gut Bacterial Community Within Galerucella Leaf Beetles.}, journal = {Microbial ecology}, volume = {86}, number = {4}, pages = {2477-2487}, pmid = {37314477}, issn = {1432-184X}, support = {#2019-4980//The Swedish Research Council Vetenskapsrådet to PAH/ ; }, mesh = {Animals ; *Coleoptera/microbiology ; Phylogeny ; *Gastrointestinal Microbiome ; RNA, Ribosomal, 16S/genetics ; Insecta ; Bacteria/genetics ; }, abstract = {Gut microbes play important roles for their hosts. Previous studies suggest that host-microbial systems can form long-term associations over evolutionary time and the dynamic changes of the intestinal system may represent major driving forces and contribute to insect dietary diversification and speciation. Our study system includes a set of six closely related leaf beetle species (Galerucella spp.) and our study aims to separate the roles of host phylogeny and ecology in determining the gut microbial community and to identify eventual relationship between host insects and gut bacteria. We collected adult beetles from their respective host plants and quantified their microbial community using 16S rRNA sequencing. The results showed that the gut bacteria community composition was structured by host beetle phylogeny, where more or less host-specific gut bacteria interact with the different Galerucella species. For example, the endosymbiotic bacteria Wolbachia was found almost exclusively in G. nymphaea and G. sagittariae. Diversity indicators also suggested that α- and β-diversities of gut bacteria communities varied among host beetle species. Overall, our results suggest a phylogenetically controlled co-occurrence pattern between the six closely related Galerucella beetles and their gut bacteria, indicating the potential of co-evolutionary processes occurring between hosts and their gut bacterial communities.}, } @article {pmid37298563, year = {2023}, author = {Fiutek, N and Couger, MB and Pirro, S and Roy, SW and de la Torre, JR and Connor, EF}, title = {Genomic Assessment of the Contribution of the Wolbachia Endosymbiont of Eurosta solidaginis to Gall Induction.}, journal = {International journal of molecular sciences}, volume = {24}, number = {11}, pages = {}, pmid = {37298563}, issn = {1422-0067}, support = {IRGEN_RG_2021-1345/IRGEN/IRGEN/United States ; }, mesh = {Animals ; *Wolbachia/genetics ; Tryptophan ; *Tephritidae/metabolism ; Insecta/metabolism ; Indoleacetic Acids/metabolism ; Cytokinins ; Genomics ; }, abstract = {We explored the genome of the Wolbachia strain, wEsol, symbiotic with the plant-gall-inducing fly Eurosta solidaginis with the goal of determining if wEsol contributes to gall induction by its insect host. Gall induction by insects has been hypothesized to involve the secretion of the phytohormones cytokinin and auxin and/or proteinaceous effectors to stimulate cell division and growth in the host plant. We sequenced the metagenome of E. solidaginis and wEsol and assembled and annotated the genome of wEsol. The wEsol genome has an assembled length of 1.66 Mbp and contains 1878 protein-coding genes. The wEsol genome is replete with proteins encoded by mobile genetic elements and shows evidence of seven different prophages. We also detected evidence of multiple small insertions of wEsol genes into the genome of the host insect. Our characterization of the genome of wEsol indicates that it is compromised in the synthesis of dimethylallyl pyrophosphate (DMAPP) and S-adenosyl L-methionine (SAM), which are precursors required for the synthesis of cytokinins and methylthiolated cytokinins. wEsol is also incapable of synthesizing tryptophan, and its genome contains no enzymes in any of the known pathways for the synthesis of indole-3-acetic acid (IAA) from tryptophan. wEsol must steal DMAPP and L-methionine from its host and therefore is unlikely to provide cytokinin and auxin to its insect host for use in gall induction. Furthermore, in spite of its large repertoire of predicted Type IV secreted effector proteins, these effectors are more likely to contribute to the acquisition of nutrients and the manipulation of the host's cellular environment to contribute to growth and reproduction of wEsol than to aid E. solidaginis in manipulating its host plant. Combined with earlier work that shows that wEsol is absent from the salivary glands of E. solidaginis, our results suggest that wEsol does not contribute to gall induction by its host.}, } @article {pmid37298356, year = {2023}, author = {Mioduchowska, M and Konecka, E and Gołdyn, B and Pinceel, T and Brendonck, L and Lukić, D and Kaczmarek, Ł and Namiotko, T and Zając, K and Zając, T and Jastrzębski, JP and Bartoszek, K}, title = {Playing Peekaboo with a Master Manipulator: Metagenetic Detection and Phylogenetic Analysis of Wolbachia Supergroups in Freshwater Invertebrates.}, journal = {International journal of molecular sciences}, volume = {24}, number = {11}, pages = {}, pmid = {37298356}, issn = {1422-0067}, support = {2017/01/X/NZ8/01873//National Science Centre/ ; 2017/27/B/NZ8/01056//National Science Centre/ ; 2021/43/D/NZ8/00344//National Science Centre/ ; 538/L260/B149/18//Young Scientists competition of University of Gdansk, Poland/ ; 1220/146/2021//UGrants-first of University of Gdansk, Poland/ ; 7862//European Molecular Biology Organization (EMBO)/ ; 2017-04951//Vetenskapsrådets Grant/ ; CA18239//COST (European Cooperation in Science and Technology)/ ; }, mesh = {Animals ; Male ; *Wolbachia/genetics ; Phylogeny ; Ecosystem ; *Arthropods ; Bacteria ; Crustacea ; Symbiosis ; }, abstract = {The infamous "master manipulators"-intracellular bacteria of the genus Wolbachia-infect a broad range of phylogenetically diverse invertebrate hosts in terrestrial ecosystems. Wolbachia has an important impact on the ecology and evolution of their host with documented effects including induced parthenogenesis, male killing, feminization, and cytoplasmic incompatibility. Nonetheless, data on Wolbachia infections in non-terrestrial invertebrates are scarce. Sampling bias and methodological limitations are some of the reasons limiting the detection of these bacteria in aquatic organisms. In this study, we present a new metagenetic method for detecting the co-occurrence of different Wolbachia strains in freshwater invertebrates host species, i.e., freshwater Arthropoda (Crustacea), Mollusca (Bivalvia), and water bears (Tardigrada) by applying NGS primers designed by us and a Python script that allows the identification of Wolbachia target sequences from the microbiome communities. We also compare the results obtained using the commonly applied NGS primers and the Sanger sequencing approach. Finally, we describe three supergroups of Wolbachia: (i) a new supergroup V identified in Crustacea and Bivalvia hosts; (ii) supergroup A identified in Crustacea, Bivalvia, and Eutardigrada hosts, and (iii) supergroup E infection in the Crustacea host microbiome community.}, } @article {pmid37293262, year = {2023}, author = {Yang, F and Li, Y and Gao, M and Xia, Q and Wang, Q and Tang, M and Zhou, X and Guo, H and Xiao, Q and Sun, L}, title = {Comparative expression profiles of carboxylesterase orthologous CXE14 in two closely related tea geometrid species, Ectropis obliqua Prout and Ectropis grisescens Warren.}, journal = {Frontiers in physiology}, volume = {14}, number = {}, pages = {1194997}, pmid = {37293262}, issn = {1664-042X}, abstract = {Insect carboxylesterases (CXEs) can be expressed in multiple tissues and play crucial roles in detoxifying xenobiotic insecticides and degrading olfactory cues. Therefore, they have been considered as an important target for development of eco-friendly insect pest management strategies. Despite extensive investigation in most insect species, limited information on CXEs in sibling moth species is currently available. The Ectropis obliqua Prout and Ectropis grisescens Warren are two closely related tea geometrid species, which share the same host of tea plant but differ in geographical distribution, sex pheromone composition, and symbiotic bacteria abundance, providing an excellent mode species for studies of functional diversity of orthologous CXEs. In this study, we focused on EoblCXE14 due to its previously reported non-chemosensory organs-biased expression. First, the EoblCXE14 orthologous gene EgriCXE14 was cloned and sequence characteristics analysis showed that they share a conserved motif and phylogenetic relationship. Quantitative real-time polymerase chain reaction (qRT-PCR) was then used to compare the expression profiles between two Ectropis spp. The results showed that EoblCXE14 was predominately expressed in E. obliqua larvae, whereas EgriCXE14 was abundant in E. grisescens at multiple developmental stages. Interestingly, both orthologous CXEs were highly expressed in larval midgut, but the expression level of EoblCXE14 in E. obliqua midgut was significantly higher than that of EgriCXE14 in E. grisescens midgut. In addition, the potential effect of symbiotic bacteria Wolbachia on the CXE14 was examined. This study is the first to provide comparative expression profiles of orthologous CXE genes in two sibling geometrid moth species and the results will help further elucidate CXEs functions and identify a potential target for tea geometrid pest control.}, } @article {pmid37291670, year = {2023}, author = {García-Rodríguez, SN and Costa-Rodríguez, N and Matos, JI and Falcón-Cordón, Y and Morchón, R and Carretón, E and Montoya-Alonso, JA}, title = {Feline heartworm disease and environmental allergens hypersensitivity: is there a link?.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {192}, pmid = {37291670}, issn = {1756-3305}, support = {PIFULPGC-2020-2-CCSALUD-2//Universidad de Las Palmas de Gran Canaria/ ; PIFULPGC-2017-CCSALUD-3//Universidad de Las Palmas de Gran Canaria/ ; TESIS2021010010//Agencia Canaria de Investigación, Innovación y Sociedad de la Información/ ; CN-240/030/158//MERCK SHARP & DOHME ANIMAL HEALTH, S.L/ ; }, mesh = {Humans ; Cats ; Animals ; *Dirofilariasis/parasitology ; Allergens ; *Dirofilaria immitis ; *Hypersensitivity/epidemiology/veterinary ; Antibodies ; *Cat Diseases/epidemiology ; }, abstract = {BACKGROUND: Cats can be infected by Dirofilaria immitis, the causative agent of heartworm disease, characterized by respiratory signs, airway hyperreactivity, remodelling and inflammation. Allergy is a multifactorial pathology, and the role of a number of helminth parasites in the development of allergies in humans and other species has been demonstrated in many studies. The aim of the present study was to verify whether cats seropositive for D. immitis present hypersensitivity to some environmental allergens.

METHODS: Blood samples were collected from 120 cats and tested for the presence of specific immunoglobulin G antibodies against D. immitis and for hypersensitivity to 20 allergens, using commercial allergen test kits.

RESULTS: Of the 120 cats tested, 72 (60.0%) were seropositive for anti-D. immitis IgG and 55 (45.8%) showed clinical signs of heartworm disease of a respiratory nature. The results of testing with the allergen kits showed that 50.8% of cats were seropositive for ≥ 1 allergens, with the most common allergens being Dermatophagoides farinae (25.8%), Dermatophagoides pteronyssinus (20.0%), Malassezia (17.5%) and Ctenocephalides felis (14.2%). The prevalence of allergies was significantly higher-by almost threefold-in cats seropositive for D. immitis (68.1% vs. 25%). There were no significant differences between the prevalence of allergic cats and presence/absence of symptoms, and the results confirmed that symptoms were not a decisive factor for the presence of allergies. The risk for developing allergies was 6.3-fold higher in cats seropositive for D. immitis than in cats that were seronegative, confirming that seropositivity for D. immitis is a risk factor.

CONCLUSIONS: Cats with confirmed heartworm can develop serious respiratory signs, potentially leading to progression to permanent lung injury and predisposing cats to hyperresponsive airway disease. Previous studies have shown that seropositivity for D. immitis and Wolbachia is related to the presence of bronchoconstriction and bronchospasm in the affected cat. The results support the suspicion that contact with D. immitis may be a risk factor for the presence of allergies.}, } @article {pmid37289079, year = {2023}, author = {Tarabai, H and Floriano, AM and Zima, J and Filová, N and Brown, JJ and Roachell, W and Smith, RL and Beatty, NL and Vogel, KJ and Nováková, E}, title = {Microbiomes of Blood-Feeding Triatomines in the Context of Their Predatory Relatives and the Environment.}, journal = {Microbiology spectrum}, volume = {11}, number = {4}, pages = {e0168123}, pmid = {37289079}, issn = {2165-0497}, mesh = {Animals ; Phylogeny ; *Triatoma ; *Trypanosoma cruzi ; *Microbiota ; Bacteria/genetics ; }, abstract = {The importance of gut microbiomes has become generally recognized in vector biology. This study addresses microbiome signatures in North American Triatoma species of public health significance (vectors of Trypanosoma cruzi) linked to their blood-feeding strategy and the natural habitat. To place the Triatoma-associated microbiomes within a complex evolutionary and ecological context, we sampled sympatric Triatoma populations, related predatory reduviids, unrelated ticks, and environmental material from vertebrate nests where these arthropods reside. Along with five Triatoma species, we have characterized microbiomes of five reduviids (Stenolemoides arizonensis, Ploiaria hirticornis, Zelus longipes, and two Reduvius species), a single soft tick species, Ornithodoros turicata, and environmental microbiomes from selected sites in Arizona, Texas, Florida, and Georgia. The microbiomes of predatory reduviids lack a shared core microbiota. As in triatomines, microbiome dissimilarities among species correlate with dominance of a single bacterial taxon. These include Rickettsia, Lactobacillus, "Candidatus Midichloria," and Zymobacter, which are often accompanied by known symbiotic genera, i.e., Wolbachia, "Candidatus Lariskella," Asaia, Gilliamella, and Burkholderia. We have further identified a compositional convergence of the analyzed microbiomes in regard to the host phylogenetic distance in both blood-feeding and predatory reduviids. While the microbiomes of the two reduviid species from the Emesinae family reflect their close relationship, the microbiomes of all Triatoma species repeatedly form a distinct monophyletic cluster highlighting their phylosymbiosis. Furthermore, based on environmental microbiome profiles and blood meal analysis, we propose three epidemiologically relevant and mutually interrelated bacterial sources for Triatoma microbiomes, i.e., host abiotic environment, host skin microbiome, and pathogens circulating in host blood. IMPORTANCE This study places microbiomes of blood-feeding North American Triatoma vectors (Reduviidae) into a broader evolutionary and ecological context provided by related predatory assassin bugs (Reduviidae), another unrelated vector species (soft tick Ornithodoros turicata), and the environment these arthropods coinhabit. For both vectors, microbiome analyses suggest three interrelated sources of bacteria, i.e., the microbiome of vertebrate nests as their natural habitat, the vertebrate skin microbiome, and the pathobiome circulating in vertebrate blood. Despite an apparent influx of environment-associated bacteria into the arthropod microbiomes, Triatoma microbiomes retain their specificity, forming a distinct cluster that significantly differs from both predatory relatives and ecologically comparable ticks. Similarly, within the related predatory Reduviidae, we found the host phylogenetic distance to underlie microbiome similarities.}, } @article {pmid37286189, year = {2023}, author = {Yang, Q and Gill, A and Robinson, KL and Umina, PA and Ross, PA and Zhan, D and Brown, C and Bell, N and MacMahon, A and Hoffmann, AA}, title = {A diversity of endosymbionts across Australian aphids and their persistence in aphid cultures.}, journal = {Environmental microbiology}, volume = {25}, number = {10}, pages = {1988-2001}, doi = {10.1111/1462-2920.16432}, pmid = {37286189}, issn = {1462-2920}, mesh = {Animals ; *Aphids/genetics/microbiology ; Symbiosis ; Australia ; Enterobacteriaceae ; Serratia/genetics ; }, abstract = {There is increasing interest in the use of endosymbionts in pest control, which will benefit from the identification of endosymbionts from potential donor species for transfer to pest species. Here, we screened for endosymbionts in 123 Australian aphid samples across 32 species using 16S DNA metabarcoding. We then developed a qPCR method to validate the metabarcoding data set and to monitor endosymbiont persistence in aphid cultures. Pea aphids (Acyrthosiphon pisum) were frequently coinfected with Rickettsiella and Serratia, and glasshouse potato aphids (Aulacorthum solani) were coinfected with Regiella and Spiroplasma; other secondary endosymbionts detected in samples occurred by themselves. Hamiltonella, Rickettsia and Wolbachia were restricted to a single aphid species, whereas Regiella was found in multiple species. Rickettsiella, Hamiltonella and Serratia were stably maintained in laboratory cultures, although others were lost rapidly. The overall incidence of secondary endosymbionts in Australian samples tended to be lower than recorded from aphids overseas. These results indicate that aphid endosymbionts probably exhibit different levels of infectivity and vertical transmission efficiency across hosts, which may contribute to natural infection patterns. The rapid loss of some endosymbionts in cultures raises questions about factors that maintain them under field conditions, while endosymbionts that persisted in laboratory culture provide candidates for interspecific transfers.}, } @article {pmid37285552, year = {2023}, author = {Hochstrasser, M}, title = {Molecular Biology of Cytoplasmic Incompatibility Caused by Wolbachia Endosymbionts.}, journal = {Annual review of microbiology}, volume = {77}, number = {}, pages = {299-316}, doi = {10.1146/annurev-micro-041020-024616}, pmid = {37285552}, issn = {1545-3251}, mesh = {Female ; Male ; Humans ; *Wolbachia/genetics ; Semen ; Reproduction/genetics ; Cytoplasm ; Molecular Biology ; Symbiosis ; }, abstract = {Among endosymbiotic bacteria living within eukaryotic cells, Wolbachia is exceptionally widespread, particularly in arthropods. Inherited through the female germline, it has evolved ways to increase the fraction of bacterially infected offspring by inducing parthenogenesis, feminization, male killing, or, most commonly, cytoplasmic incompatibility (CI). In CI, Wolbachia infection of males causes embryonic lethality unless they mate with similarly infected females, creating a relative reproductive advantage for infected females. A set of related Wolbachia bicistronic operons encodes the CI-inducing factors. The downstream gene encodes a deubiquitylase or nuclease and is responsible for CI induction by males, while the upstream product when expressed in females binds its sperm-introduced cognate partner and rescues viability. Both toxin-antidote and host-modification mechanisms have been proposed to explain CI. Interestingly, male killing by either Spiroplasma or Wolbachia endosymbionts involves deubiquitylases as well. Interference with the host ubiquitin system may therefore be a common theme among endosymbiont-mediated reproductive alterations.}, } @article {pmid37278210, year = {2023}, author = {Xu, X and Hoffmann, AA and Umina, PA and Ward, SE and Coquilleau, MP and Malipatil, MB and Ridland, PM}, title = {Molecular identification of hymenopteran parasitoids and their endosymbionts from agromyzids.}, journal = {Bulletin of entomological research}, volume = {113}, number = {4}, pages = {481-496}, doi = {10.1017/S0007485323000160}, pmid = {37278210}, issn = {1475-2670}, support = {MT20005//Hort Innovation/ ; }, mesh = {Animals ; Phylogeny ; *Wasps/genetics ; *Diptera/genetics ; Australia ; Crops, Agricultural ; DNA ; }, abstract = {Three polyphagous pest Liriomyza spp. (Diptera: Agromyzidae) have recently invaded Australia and are damaging horticultural crops. Parasitic wasps are recognized as effective natural enemies of leafmining species globally and are expected to become important biocontrol agents in Australia. However, the hymenopteran parasitoid complex of agromyzids in Australia is poorly known and its use hindered due to taxonomic challenges when based on morphological characters. Here, we identified 14 parasitoid species of leafminers based on molecular and morphological data. We linked DNA barcodes (5' end cytochrome c oxidase subunit I (COI) sequences) to five adventive eulophid wasp species (Chrysocharis pubicornis (Zetterstedt), Diglyphus isaea (Walker), Hemiptarsenus varicornis (Girault), Neochrysocharis formosa (Westwood), and Neochrysocharis okazakii Kamijo) and two braconid species (Dacnusa areolaris (Nees) and Opius cinerariae Fischer). We also provide the first DNA barcodes (5' end COI sequences) with linked morphological characters for seven wasp species, with three identified to species level (Closterocerus mirabilis Edwards & La Salle, Trigonogastrella parasitica (Girault), and Zagrammosoma latilineatum Ubaidillah) and four identified to genus (Aprostocetus sp., Asecodes sp., Opius sp. 1, and Opius sp. 2). Phylogenetic analyses suggest C. pubicornis, D. isaea, H. varicornis, and O. cinerariae are likely cryptic species complexes. Neochrysocharis formosa and Aprostocetus sp. specimens were infected with Rickettsia. Five other species (Cl. mirabilis, D. isaea, H. varicornis, Opius sp. 1, and Opius sp. 2) were infected with Wolbachia, while two endosymbionts (Rickettsia and Wolbachia) co-infected N. okazakii. These findings provide background information about the parasitoid fauna expected to help control the leafminers.}, } @article {pmid37270375, year = {2023}, author = {Chaisiri, K and Linsuwanon, P and Makepeace, BL}, title = {The chigger microbiome: big questions in a tiny world.}, journal = {Trends in parasitology}, volume = {39}, number = {8}, pages = {696-707}, doi = {10.1016/j.pt.2023.05.002}, pmid = {37270375}, issn = {1471-5007}, mesh = {Humans ; Animals ; *Trombiculidae/microbiology ; *Orientia tsutsugamushi ; *Scrub Typhus/microbiology ; *Rickettsia ; Zoonoses ; }, abstract = {'Chiggers' (trombiculid mite larvae) are best known as vectors of rickettsial pathogens, Orientia spp., which cause a zoonosis, scrub typhus. However, several other pathogens (e.g., Hantaan orthohantavirus, Dabie bandavirus, Anaplasma spp., Bartonella spp., Borrelia spp., and Rickettsia spp.) and bacterial symbionts (e.g., Cardinium, Rickettsiella, and Wolbachia) are being reported from chiggers with increasing frequency. Here, we explore the surprisingly diverse chigger microbiota and potential interactions within this microcosm. Key conclusions include a possible role for chiggers as vectors of viral diseases; the dominance in some chigger populations of unidentified symbionts in several bacterial families; and increasing evidence for vertical transmission of potential pathogens and symbiotic bacteria in chiggers, suggesting intimate interactions and not simply incidental acquisition of bacteria from the environment or host.}, } @article {pmid37269018, year = {2023}, author = {Peña-Espinoza, M and Em, D and Shahi-Barogh, B and Berer, D and Duscher, GG and van der Vloedt, L and Glawischnig, W and Rehbein, S and Harl, J and Unterköfler, MS and Fuehrer, HP}, title = {Molecular pathogen screening of louse flies (Diptera: Hippoboscidae) from domestic and wild ruminants in Austria.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {179}, pmid = {37269018}, issn = {1756-3305}, support = {Austrian Barcode of Life - Hochschulraum-Strukturmittel//Austrian Federal Ministry of Education, Science and Research/ ; }, mesh = {Humans ; Animals ; Sheep ; Cattle ; *Diptera ; *Deer/parasitology ; Austria/epidemiology ; Phylogeny ; Ruminants ; *Bartonella/genetics ; *Anoplura ; *Anaplasmataceae/genetics ; *Piroplasmida ; }, abstract = {BACKGROUND: Hippoboscid flies (Diptera: Hippoboscidae), also known as louse flies or keds, are obligate blood-sucking ectoparasites of animals, and accidentally of humans. The potential role of hippoboscids as vectors of human and veterinary pathogens is being increasingly investigated, but the presence and distribution of infectious agents in louse flies is still unknown in parts of Europe. Here, we report the use of molecular genetics to detect and characterize vector-borne pathogens in hippoboscid flies infesting domestic and wild animals in Austria.

METHODS: Louse flies were collected from naturally infested cattle (n = 25), sheep (n = 3), and red deer (n = 12) across Austria between 2015 and 2019. Individual insects were morphologically identified to species level and subjected to DNA extraction for molecular pathogen screening and barcoding. Genomic DNA from each louse fly was screened for Borrelia spp., Bartonella spp., Trypanosomatida, Anaplasmataceae, Filarioidea and Piroplasmida. Obtained sequences of Trypanosomatida and Bartonella spp. were further characterized by phylogenetic and haplotype networking analyses.

RESULTS: A total of 282 hippoboscid flies corresponding to three species were identified: Hippobosca equina (n = 62) collected from cattle, Melophagus ovinus (n = 100) from sheep and Lipoptena cervi (n = 120) from red deer (Cervus elaphus). Molecular screening revealed pathogen DNA in 54.3% of hippoboscids, including infections with single (63.39%), two (30.71%) and up to three (5.90%) distinct pathogens in the same individual. Bartonella DNA was detected in 36.9% of the louse flies. Lipoptena cervi were infected with 10 distinct and previously unreported Bartonella sp. haplotypes, some closely associated with strains of zoonotic potential. DNA of trypanosomatids was identified in 34% of hippoboscids, including the first description of Trypanosoma sp. in H. equina. Anaplasmataceae DNA (Wolbachia spp.) was detected only in M. ovinus (16%), while < 1% of the louse flies were positive for Borrelia spp. and Filarioidea. All hippoboscids were negative for Piroplasmida.

CONCLUSIONS: Molecular genetic screening confirmed the presence of several pathogens in hippoboscids infesting domestic and wild ruminants in Austria, including novel pathogen haplotypes of zoonotic potential (e.g. Bartonella spp.) and the first report of Trypanosoma sp. in H. equina, suggesting a potential role of this louse fly as vector of animal trypanosomatids. Experimental transmission studies and expanded monitoring of hippoboscid flies and hippoboscid-associated pathogens are warranted to clarify the competence of these ectoparasites as vectors of infectious agents in a One-Health context.}, } @article {pmid37266040, year = {2023}, author = {Rodrigues, J and Lefoulon, E and Gavotte, L and Perillat-Sanguinet, M and Makepeace, B and Martin, C and D'Haese, CA}, title = {Wolbachia springs eternal: symbiosis in Collembola is associated with host ecology.}, journal = {Royal Society open science}, volume = {10}, number = {5}, pages = {230288}, pmid = {37266040}, issn = {2054-5703}, abstract = {Wolbachia are endosymbiotic alpha-proteobacteria infecting a wide range of arthropods and nematode hosts with diverse interactions, from reproductive parasites to obligate mutualists. Their taxonomy is defined by lineages called supergroups (labelled by letters of the alphabet), while their evolutionary history is complex, with multiple horizontal transfers and secondary losses. One of the least recently derived, supergroup E, infects springtails (Collembola), widely distributed hexapods, with sexual and/or parthenogenetic populations depending on species. To better characterize the diversity of Wolbachia infecting springtails, the presence of Wolbachia was screened in 58 species. Eleven (20%) species were found to be positive, with three Wolbachia genotypes identified for the first time in supergroup A. The novel genotypes infect springtails ecologically and biologically different from those infected by supergroup E. To root the Wolbachia phylogeny, rather than distant other Rickettsiales, supergroup L infecting plant-parasitic nematodes was used here. We hypothesize that the ancestor of Wolbachia was consumed by soil-dwelling nematodes, and was transferred horizontally via plants into aphids, which then infected edaphic arthropods (e.g. springtails and oribatid mites) before expanding into most clades of terrestrial arthropods and filarial nematodes.}, } @article {pmid37254297, year = {2023}, author = {Matsufuji, T and Seirin-Lee, S}, title = {The optimal strategy of incompatible insect technique (IIT) using Wolbachia and the application to malaria control.}, journal = {Journal of theoretical biology}, volume = {569}, number = {}, pages = {111519}, doi = {10.1016/j.jtbi.2023.111519}, pmid = {37254297}, issn = {1095-8541}, mesh = {Animals ; Male ; Humans ; Female ; *Aedes ; *Wolbachia ; Mosquito Vectors ; Reproduction ; Mosquito Control/methods ; *Malaria/prevention & control ; }, abstract = {For decades, techniques to control vector population with low environmental impact have been widely explored in both field and theoretical studies. The incompatible insect technique (IIT) using Wolbachia, based on cytoplasmic incompatibility, is a technique that Wolbachia-infected male mosquitoes are incapable of producing viable offspring after mating with wild-type female mosquitoes. While the IIT method experimentally ensured its effectiveness in several field works, the failure of female mosquito population control by replacement owing to the accidental contamination of Wolbachia-infected female mosquitoes has been a concern and an obstacle in implementing the IIT method in nature. In this study, we develop a population-based IIT mathematical model using cytoplasmic incompatibility and evaluate the effectiveness of the IIT method in scenarios where contamination is present or absent. In addition, by extending the model to assess the disease infection status of the human population with malaria, we evaluate the optimal release strategy and cost for successful disease control. Our study proves that IIT could be a promising method to control mosquito-borne diseases without perfect suppression of vector mosquito population regardless of contamination.}, } @article {pmid37253037, year = {2023}, author = {Turner, HC and Quyen, DL and Dias, R and Huong, PT and Simmons, CP and Anders, KL}, title = {An economic evaluation of Wolbachia deployments for dengue control in Vietnam.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {5}, pages = {e0011356}, pmid = {37253037}, issn = {1935-2735}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Humans ; Cost-Benefit Analysis ; *Dengue/epidemiology/prevention & control ; *Wolbachia ; Vietnam/epidemiology ; Mosquito Vectors ; *Aedes/microbiology ; }, abstract = {INTRODUCTION: Dengue is a major public health challenge and a growing problem due to climate change. The release of Aedes aegypti mosquitoes infected with the intracellular bacterium Wolbachia is a novel form of vector control against dengue. However, there remains a need to evaluate the benefits of such an intervention at a large scale. In this paper, we evaluate the potential economic impact and cost-effectiveness of scaled Wolbachia deployments as a form of dengue control in Vietnam-targeted at the highest burden urban areas.

METHODS: Ten settings within Vietnam were identified as priority locations for potential future Wolbachia deployments (using a population replacement strategy). The effectiveness of Wolbachia deployments in reducing the incidence of symptomatic dengue cases was assumed to be 75%. We assumed that the intervention would maintain this effectiveness for at least 20 years (but tested this assumption in the sensitivity analysis). A cost-utility analysis and cost-benefit analysis were conducted.

RESULTS: From the health sector perspective, the Wolbachia intervention was projected to cost US$420 per disability-adjusted life year (DALY) averted. From the societal perspective, the overall cost-effectiveness ratio was negative, i.e. the economic benefits outweighed the costs. These results are contingent on the long-term effectiveness of Wolbachia releases being sustained for 20 years. However, the intervention was still classed as cost-effective across the majority of the settings when assuming only 10 years of benefits.

CONCLUSION: Overall, we found that targeting high burden cities with Wolbachia deployments would be a cost-effective intervention in Vietnam and generate notable broader benefits besides health gains.}, } @article {pmid37250803, year = {2023}, author = {Arai, H and Anbutsu, H and Nishikawa, Y and Kogawa, M and Ishii, K and Hosokawa, M and Lin, SR and Ueda, M and Nakai, M and Kunimi, Y and Harumoto, T and Kageyama, D and Takeyama, H and Inoue, MN}, title = {Combined actions of bacteriophage-encoded genes in Wolbachia-induced male lethality.}, journal = {iScience}, volume = {26}, number = {6}, pages = {106842}, pmid = {37250803}, issn = {2589-0042}, abstract = {Some Wolbachia endosymbionts induce male killing, whereby male offspring of infected females are killed during development; however, the origin and diversity of the underlying mechanisms remain unclear. In this study, we identified a 76 kbp prophage region specific to male-killing Wolbachia hosted by the moth Homona magnanima. The prophage encoded a homolog of the male-killing gene oscar in Ostrinia moths and the wmk gene that induces various toxicities in Drosophila melanogaster. Upon overexpressing these genes in D. melanogaster, wmk-1 and wmk-3 killed all males and most females, whereas Hm-oscar, wmk-2, and wmk-4 had no impact on insect survival. Strikingly, co-expression of tandemly arrayed wmk-3 and wmk-4 killed 90% of males and restored 70% of females, suggesting their conjugated functions for male-specific lethality. While the male-killing gene in the native host remains unknown, our findings highlight the role of bacteriophages in male-killing evolution and differences in male-killing mechanisms among insects.}, } @article {pmid37247378, year = {2023}, author = {Oladipupo, SO and Laidoudi, Y and Beckmann, JF and Hu, XP and Appel, AG}, title = {The prevalence of Wolbachia in multiple cockroach species and its implication for urban insect management.}, journal = {Journal of economic entomology}, volume = {116}, number = {4}, pages = {1307-1316}, doi = {10.1093/jee/toad098}, pmid = {37247378}, issn = {1938-291X}, mesh = {Animals ; *Wolbachia/genetics ; Biotin/genetics ; Phylogeny ; Prevalence ; Insecta ; *Bedbugs ; *Blattellidae ; Symbiosis ; }, abstract = {Cockroach management relies heavily on the use of conventional insecticides in urban settings, which no longer provide the anticipated level of control. Knowledge of cockroach endosymbionts, like Wolbachia, might provide novel avenues for control. Therefore, we screened 16 cockroach species belonging to 3 families (Ectobiidae, Blattidae, and Blaberidae) for the presence of Wolbachia. We mapped the evolution of Wolbachia-cockroach relationships based on maximum likelihood phylogeny and phylogenetic species clustering on a multi-loci sequence dataset (i.e., coxA, virD4, hcpA, and gatB) of Wolbachia genes. We confirmed the previous report of Wolbachia in 1 Ectobiid species; Supella longipalpa (Fab.), and detected the presence of Wolbachia in 2 Ectobiid species; Balta notulata (Stål) and Pseudomops septentrionalis Hebard, and 1 Blaberid species; Gromphadorhina portentosa (Schaum). All cockroach-associated Wolbachia herein detected were clustered with the ancestor of F clade Wolbachia of Cimex lectularius L. (bed bugs). Since Wolbachia provision C. lectularius with biotin vitamins that confer reproductive fitness, we screened the cockroach-associated Wolbachia for the presence of biotin genes. In toto, our results reveal 2 important findings: (i) Wolbachia is relatively uncommon among cockroach species infecting about 25% of species investigated, and (ii) cockroach-associated Wolbachia have biotin genes that likely provide nutritional benefits to their hosts. Thus, we discuss the potential of exploring Wolbachia as a tool for urban insect management.}, } @article {pmid37231184, year = {2023}, author = {Zheng, R and Wang, Q and Wu, R and Paradkar, PN and Hoffmann, AA and Wang, GH}, title = {Holobiont perspectives on tripartite interactions among microbiota, mosquitoes, and pathogens.}, journal = {The ISME journal}, volume = {17}, number = {8}, pages = {1143-1152}, pmid = {37231184}, issn = {1751-7370}, mesh = {Animals ; Humans ; *Culicidae ; Mosquito Vectors ; *Microbiota/physiology ; *Malaria ; *Wolbachia/physiology ; }, abstract = {Mosquito-borne diseases like dengue and malaria cause a significant global health burden. Unfortunately, current insecticides and environmental control strategies aimed at the vectors of these diseases are only moderately effective in decreasing disease burden. Understanding and manipulating the interaction between the mosquito holobiont (i.e., mosquitoes and their resident microbiota) and the pathogens transmitted by these mosquitoes to humans and animals could help in developing new disease control strategies. Different microorganisms found in the mosquito's microbiota affect traits related to mosquito survival, development, and reproduction. Here, we review the physiological effects of essential microbes on their mosquito hosts; the interactions between the mosquito holobiont and mosquito-borne pathogen (MBP) infections, including microbiota-induced host immune activation and Wolbachia-mediated pathogen blocking (PB); and the effects of environmental factors and host regulation on the composition of the microbiota. Finally, we briefly overview future directions in holobiont studies, and how these may lead to new effective control strategies against mosquitoes and their transmitted diseases.}, } @article {pmid37231093, year = {2023}, author = {Bruner-Montero, G and Jiggins, FM}, title = {Wolbachia protects Drosophila melanogaster against two naturally occurring and virulent viral pathogens.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {8518}, pmid = {37231093}, issn = {2045-2322}, mesh = {Animals ; Female ; Drosophila melanogaster ; *Wolbachia ; *Viruses ; *RNA Viruses ; Antiviral Agents/pharmacology ; Symbiosis ; }, abstract = {Wolbachia is a common endosymbiont that can protect insects against viral pathogens. However, whether the antiviral effects of Wolbachia have a significant effect on fitness remains unclear. We have investigated the interaction between Drosophila melanogaster, Wolbachia and two viruses that we recently isolated from wild flies, La Jolla virus (LJV; Iflaviridae) and Newfield virus (NFV; Permutotetraviridae). Flies infected with these viruses have increased mortality rates, and NFV partially sterilizes females. These effects on fitness were reduced in Wolbachia-infected flies, and this was associated with reduced viral titres. However, Wolbachia alone also reduces survival, and under our experimental conditions these costs of the symbiont can outweigh the benefits of antiviral protection. In contrast, protection against the sterilizing effect of NFV leads to a net benefit of Wolbachia infection after exposure to the virus. These results support the hypothesis that Wolbachia is an important defense against the natural pathogens of D. melanogaster. Furthermore, by reducing the cost of Wolbachia infection, the antiviral effects of Wolbachia may aid its invasion into populations and help explain why it is so common in nature.}, } @article {pmid37214959, year = {2023}, author = {Moldovan, OT and Carrell, AA and Bulzu, PA and Levei, E and Bucur, R and Sitar, C and Faur, L and Mirea, IC and Enilă, M and Cadar, O and Podar, M}, title = {The gut microbiome mediates adaptation to scarce food in Coleoptera.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2023.05.12.540564}, pmid = {37214959}, abstract = {Beetles are ubiquitous cave invertebrates worldwide that adapted to scarce subterranean resources when they colonized caves. Here, we investigated the potential role of gut microbiota in the adaptation of beetles to caves from different climatic regions of the Carpathians. The beetles' microbiota was host-specific, reflecting phylogenetic and nutritional adaptation. The microbial community structure further resolved conspecific beetles by caves suggesting microbiota-host coevolution and influences by local environmental factors. The detritivore species hosted a variety of bacteria known to decompose and ferment organic matter, suggesting turnover and host cooperative digestion of the sedimentary microbiota and allochthonous-derived nutrients. The cave Carabidae, with strong mandibulae adapted to predation and scavenging of animal and plant remains, had distinct microbiota dominated by symbiotic lineages Spiroplasma or Wolbachia . All beetles had relatively high levels of fermentative Carnobacterium and Vagococcus involved in lipid accumulation and a reduction of metabolic activity, and both features characterize adaptation to caves.}, } @article {pmid37214831, year = {2023}, author = {Mfopit, YM and Weber, JS and Chechet, GD and Ibrahim, MAM and Signaboubo, D and Achukwi, DM and Mamman, M and Balogun, EO and Shuaibu, MN and Kabir, J and Kelm, S}, title = {Molecular detection of Sodalis glossinidius, Spiroplasma and Wolbachia endosymbionts in wild population of tsetse flies collected in Cameroon, Chad and Nigeria.}, journal = {Research square}, volume = {}, number = {}, pages = {}, pmid = {37214831}, support = {K43 TW012015/TW/FIC NIH HHS/United States ; }, abstract = {Background Tsetse flies are cyclical vectors of African trypanosomiasis. They have established symbiotic associations with different bacteria, which influence certain aspects of their physiology. The vector competence of tsetse flies for different trypanosome species is highly variable and is suggested to be affected by various factors, amongst which are bacterial endosymbionts. Symbiotic interactions may provide an avenue for the disease control. The current study provided the prevalence of 3 tsetse symbionts in Glossina species from Cameroon, Chad and Nigeria. Results Tsetse flies were collected from five different locations and dissected. DNA was extracted and polymerase chain reaction PCR was used to detect the presence of Sodalis glossinidius , Spiroplasma sp and Wolbachia using specific primers. A total of 848 tsetse samples were analysed: Glossina morsitans submorsitans (47.52%), Glossina palpalis palpalis (37.26%), Glossina fuscipes fuscipes (9.08%) and Glossina tachinoides (6.13%). Only 95 (11.20%) were infected with at least one of the 3 symbionts. Among the infected, 6 (6.31%) were carrying mixed infection (Wolbachia and Spiroplasma). The overall symbiont prevalence was 0.88%, 3.66% and 11.00% respectively, for Sodalis , Spiroplasma and Wolbachia . Prevalence varied between countries and tsetse species. No Spiroplasma was detected in samples from Cameroon and no Sodalis was found in samples from Nigeria. Conclusion The present study revealed for the first time, the presence of infection by Spiroplasma in tsetse in Chad and Nigeria. These findings provide useful information to the repertoire of bacterial flora of tsetse flies and incite to more investigations to understand their implication in the vector competence of tsetse flies.}, } @article {pmid37213490, year = {2023}, author = {Jackson, R and Patapiou, PA and Golding, G and Helanterä, H and Economou, CK and Chapuisat, M and Henry, LM}, title = {Evidence of phylosymbiosis in Formica ants.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1044286}, pmid = {37213490}, issn = {1664-302X}, abstract = {INTRODUCTION: Insects share intimate relationships with microbes that play important roles in their biology. Yet our understanding of how host-bound microbial communities assemble and perpetuate over evolutionary time is limited. Ants host a wide range of microbes with diverse functions and are an emerging model for studying the evolution of insect microbiomes. Here, we ask whether phylogenetically related ant species have formed distinct and stable microbiomes.

METHODS: To answer this question, we investigated the microbial communities associated with queens of 14 Formica species from five clades, using deep coverage 16S rRNA amplicon sequencing.

RESULTS: We reveal that Formica species and clades harbor highly defined microbial communities that are dominated by four bacteria genera: Wolbachia, Lactobacillus, Liliensternia, and Spiroplasma. Our analysis reveals that the composition of Formica microbiomes mirrors the phylogeny of the host, i.e., phylosymbiosis, in that related hosts harbor more similar microbial communities. In addition, we find there are significant correlations between microbe co-occurrences.

DISCUSSION: Our results demonstrate Formica ants carry microbial communities that recapitulate the phylogeny of their hosts. Our data suggests that the co-occurrence of different bacteria genera may at least in part be due to synergistic and antagonistic interactions between microbes. Additional factors potentially contributing to the phylosymbiotic signal are discussed, including host phylogenetic relatedness, host-microbe genetic compatibility, modes of transmission, and similarities in host ecologies (e.g., diets). Overall, our results support the growing body of evidence that microbial community composition closely depends on the phylogeny of their hosts, despite bacteria having diverse modes of transmission and localization within the host.}, } @article {pmid37196858, year = {2023}, author = {Ward, MCE and Barrios, MC and Fallon, AM}, title = {Paraquat is toxic to the soil-dwelling arthropod, Folsomia candida (Collembola: Isotomidae), and has potential effects on its Wolbachia endosymbiont.}, journal = {Journal of invertebrate pathology}, volume = {198}, number = {}, pages = {107936}, doi = {10.1016/j.jip.2023.107936}, pmid = {37196858}, issn = {1096-0805}, mesh = {Female ; Animals ; *Arthropods ; Paraquat ; *Wolbachia ; Soil ; Charcoal ; Reproduction ; }, abstract = {The springtail, Folsomia candida, is a soil arthropod commonly used to evaluate environmental toxins. Conflicting data on the toxicity of the herbicide paraquat prompted re-evaluation of its effects on F. candida survival and reproduction. Paraquat has an LC50 of about 80 μM when tested in the absence of charcoal; charcoal, often used in test arenas to facilitate visualization of the white Collembola, has a protective effect. Survivors of paraquat treatment fail to resume molting and oviposition, suggesting an irreversible effect on the Wolbachia symbiont that restores diploidy during parthenogenetic reproduction of this species.}, } @article {pmid37194361, year = {2023}, author = {Li, J and Dong, B and Zhong, Y and Li, ZX}, title = {Transinfected Wolbachia strains induce a complex of cytoplasmic incompatibility phenotypes: Roles of CI factor genes.}, journal = {Environmental microbiology reports}, volume = {15}, number = {5}, pages = {370-382}, pmid = {37194361}, issn = {1758-2229}, mesh = {Animals ; Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Cytoplasm ; Base Sequence ; Phenotype ; *Hemiptera ; }, abstract = {Wolbachia can modulate the reproductive development of their hosts in multiple modes, and cytoplasmic incompatibility (CI) is the most well-studied phenotype. The whitefly Bemisia tabaci is highly receptive to different Wolbachia strains: wCcep strain from the rice moth Corcyra cephalonica and wMel strain from the fruit fly Drosophila melanogaster could successfully establish and induce CI in transinfected whiteflies. Nevertheless, it is unknown what will happen when these two exogenous Wolbachia strains are co-transinfected into a new host. Here, we artificially transinferred wCcep and wMel into the whitefly and established double- and singly-transinfected B. tabaci isofemale lines. Reciprocal crossing experiments showed that wCcep and wMel induced a complex of CI phenotypes in the recipient host, including unidirectional and bidirectional CI. We next sequenced the whole genome of wCcep and performed a comparative analysis of the CI factor genes between wCcep and wMel, indicating that their cif genes were phylogenetically and structurally divergent, which can explain the crossing results. The amino acid sequence identity and structural features of Cif proteins may be useful parameters for predicting their function. Structural comparisons between CifA and CifB provide valuable clues for explaining the induction or rescue of CI observed in crossing experiments between transinfected hosts.}, } @article {pmid37186167, year = {2023}, author = {Shi, H and Yu, X and Cheng, G}, title = {Impact of the microbiome on mosquito-borne diseases.}, journal = {Protein & cell}, volume = {14}, number = {10}, pages = {743-761}, pmid = {37186167}, issn = {1674-8018}, support = {2021YFC2300200//National Key R&D Program of China/ ; 32188101//National Natural Science Foundation of China/ ; 202005AF150034//Yunnan Chenggong expert workstation/ ; 202105AE160020//Innovation Team Project of Yunnan Science and Technology Department/ ; 2022THFS6124//Tsinghua-Foshan Innovation Special Fund/ ; }, mesh = {Animals ; Humans ; *Culicidae/microbiology ; *Vector Borne Diseases ; *Gastrointestinal Microbiome ; *Wolbachia ; }, abstract = {Mosquito-borne diseases present a significant threat to human health, with the possibility of outbreaks of new mosquito-borne diseases always looming. Unfortunately, current measures to combat these diseases such as vaccines and drugs are often either unavailable or ineffective. However, recent studies on microbiomes may reveal promising strategies to fight these diseases. In this review, we examine recent advances in our understanding of the effects of both the mosquito and vertebrate microbiomes on mosquito-borne diseases. We argue that the mosquito microbiome can have direct and indirect impacts on the transmission of these diseases, with mosquito symbiotic microorganisms, particularly Wolbachia bacteria, showing potential for controlling mosquito-borne diseases. Moreover, the skin microbiome of vertebrates plays a significant role in mosquito preferences, while the gut microbiome has an impact on the progression of mosquito-borne diseases in humans. As researchers continue to explore the role of microbiomes in mosquito-borne diseases, we highlight some promising future directions for this field. Ultimately, a better understanding of the interplay between mosquitoes, their hosts, pathogens, and the microbiomes of mosquitoes and hosts may hold the key to preventing and controlling mosquito-borne diseases.}, } @article {pmid37176154, year = {2023}, author = {Guo, W and Zhang, M and Lin, L and Zeng, C and Zhang, Y and He, X}, title = {Bacterial Community Survey of Wolbachia-Infected Parthenogenetic Parasitoid Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) Treated with Antibiotics and High Temperature.}, journal = {International journal of molecular sciences}, volume = {24}, number = {9}, pages = {}, pmid = {37176154}, issn = {1422-0067}, support = {2022KJ113//Innovation Team Project of Modern Agricultural Industrial Technology System of Guangdong Province/ ; }, mesh = {Animals ; Male ; *Hymenoptera ; Anti-Bacterial Agents/pharmacology ; Temperature ; *Wolbachia/genetics ; Parthenogenesis ; *Wasps/microbiology ; }, abstract = {Wolbachia has been shown to induce thelytokous parthenogenesis in Trichogramma species, which have been widely used as biological control agents around the world. Little is known about the changes of bacterial community after restoring arrhenotokous or bisexual reproduction in the T. pretiosum. Here, we investigate the emergence of males of T. pretiosum through curing experiments (antibiotics and high temperature), crossing experiments, and high-throughput 16S ribosomal RNA sequencing (rRNA-seq). The results of curing experiments showed that both antibiotics and high temperatures could cause the thelytokous T. pretiosum to produce male offspring. Wolbachia was dominant in the thelytokous T. pretiosum bacterial community with 99.01% relative abundance. With the relative abundance of Wolbachia being depleted by antibiotics, the diversity and relative content of other endosymbiotic bacteria increased, and the reproductive mode reverted from thelytoky to arrhenotoky in T. pretiosum. Although antibiotics did not eliminate Wolbachia in T. pretiosum, sulfadiazine showed an advantage in restoring entirely arrhenotokous and successive bisexual reproduction. This study was the first to demonstrate the bacterial communities in parthenogenetic Trichogramma before and after antibiotics or high-temperature treatment. Our findings supported the hypothesis that Wolbachia titer-dependence drives a reproduction switch in T. pretiosum between thelytoky and arrhenotoky.}, } @article {pmid37156379, year = {2023}, author = {Chang, G and Xue, H and Ji, J and Wang, L and Zhu, X and Zhang, K and Li, D and Gao, X and Niu, L and Gao, M and Luo, J and Cui, J}, title = {Risk assessment of predatory lady beetle Propylea japonica's multi-generational exposure to three non-insecticidal agrochemicals.}, journal = {The Science of the total environment}, volume = {886}, number = {}, pages = {163931}, doi = {10.1016/j.scitotenv.2023.163931}, pmid = {37156379}, issn = {1879-1026}, mesh = {Female ; Male ; Animals ; Bees ; *Coleoptera/physiology ; Bacteria ; Risk Assessment ; }, abstract = {The effects of non-insecticidal agrochemicals on pest natural predators remain largely unexplored except bees and silkworm. The herbicide quizalofop-p-ethyl (QpE), fungicide thiophanate-methyl (TM), and plant growth regulator mepiquat chloride (MC) have been extensively applied as non-insecticidal agrochemicals. Here, we systematically evaluated multiple effects of these 3 non-insecticidal agrochemicals on three generations of Propylea japonica, an important agroforestry predatory beetle, including the effects on its development, reproduction, enterobacteria, and transcriptomic response. The results showed that QpE exhibited a hormetic effect on P. japonica, thus significantly increasing the survival rate of generation 2 (F2) females, generation 3 (F3) females, and F3 males and body weight of F3 males. However, three successive generations exposed to TM and MC had no significant effect on longevity, body weight, survival rate, pre-oviposition period, and fecundity of P. japonica. Additionally, we investigated the effects of MC, TM, and QpE exposure on gene expression and gut bacterial community of F3 P. japonica. Under MC, TM, and QpE exposure, the overwhelming genes of P. japonica (99.90 %, 99.45 %, and 99.7 %) remained unaffected, respectively. Under TM and MC exposure, differentially expressed genes (DEGs) were not significantly enriched in any KEGG pathway, indicating TM and MC did not significantly affect functions of P. japonica, but under QpE exposure, the expression levels of drug metabolism-related genes were down-regulated. Although QpE treatment did not affect gut dominant bacterial community composition, it significantly increased relative abundances of detoxification metabolism-related bacteria such as Wolbachia, Pseudomonas and Burkholderia in P. japonica. However, TM and MC had no significant effect on the gut bacterial community composition and relative abundance in P. japonica. This study revealed for the first time the mechanism by which P. japonica might compensate for gene downregulation-induced detoxification metabolism decline through altering symbiotic bacteria under QpE exposure. Our findings provide reference for the rational application of non-insecticidal agrochemicals.}, } @article {pmid37154102, year = {2023}, author = {Sinha, A and Li, Z and Poole, CB and Ettwiller, L and Lima, NF and Ferreira, MU and Fombad, FF and Wanji, S and Carlow, CKS}, title = {Multiple lineages of nematode-Wolbachia symbiosis in supergroup F and convergent loss of bacterioferritin in filarial Wolbachia.}, journal = {Genome biology and evolution}, volume = {15}, number = {5}, pages = {}, pmid = {37154102}, issn = {1759-6653}, abstract = {The intracellular endosymbiotic proteobacteria Wolbachia have evolved across the phyla nematoda and arthropoda. In Wolbachia phylogeny, supergroup F is the only clade known so far with members from both arthropod and filarial nematode hosts and therefore can provide unique insights into their evolution and biology. In this study, 4 new supergroup F Wolbachia genomes have been assembled using a metagenomic assembly and binning approach, wMoz and wMpe from the human filarial parasites Mansonella ozzardi and Mansonella perstans, and wOcae and wMoviF from the blue mason bee Osmia caerulescens and the sheep ked Melophagus ovinus respectively. A comprehensive phylogenomic analysis revealed two distinct lineages of filarial Wolbachia in supergroup F, indicating multiple horizontal transfer events between arthropod and nematode hosts. The analysis also reveals that the evolution of Wolbachia-filaria symbioses is accompanied by a convergent pseudogenization and loss of the bacterioferritin gene, a phenomenon found to be shared by all filarial Wolbachia, even those outside supergroup F. These observations indicate that differences in heme metabolism might be a key feature distinguishing filarial and arthropod Wolbachia. The new genomes provide a valuable resource for further studies on symbiosis, evolution, and the discovery of new antibiotics to treat mansonellosis.}, } @article {pmid37138629, year = {2023}, author = {Tan, Y and Gong, B and Zhang, Q and Li, C and Weng, J and Zhou, X and Jin, L}, title = {Diversity of endosymbionts in camellia spiny whitefly, Aleurocanthus camelliae (Hemiptera: Aleyrodidae), estimated by 16S rRNA analysis and their biological implications.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1124386}, pmid = {37138629}, issn = {1664-302X}, abstract = {Camellia spiny whitefly, Aleurocanthus camelliae (Hemiptera: Aleyrodidae), is a major pest in tea, which poses a serious threat to tea production. Similar to many insects, various bacterial symbioses inside A. camelliae may participate in the reproduction, metabolism, and detoxification of the host. However, few reports included research on the microbial composition and influence on A. camelliae growth. We first applied high-throughput sequencing of the V4 region in the 16S rRNA of symbiotic bacteria to study its component and effect on the biological trait of A. camelliae by comparing it with the antibiotic treatment group. The population parameters, survival rate, and fecundity rate of A. camelliae were also analyzed using the age-stage two-sex life table. Our results demonstrated that phylum Proteobacteria (higher than 96.15%) dominated the whole life cycle of A. camelliae. It unveiled the presence of Candidatus Portiera (primary endosymbiont) (67.15-73.33%), Arsenophonus (5.58-22.89%), Wolbachia (4.53-11.58%), Rickettsia (0.75-2.59%), and Pseudomonas (0.99-1.88%) genus. Antibiotic treatment caused a significant decrease in the endosymbiont, which negatively affected the host's biological properties and life process. For example, 1.5% rifampicin treatment caused a longer preadult stage in the offspring generation (55.92 d) compared to the control (49.75d) and a lower survival rate (0.36) than the control (0.60). The decreased intrinsic rate of increase (r), net reproductive rate (R 0), and prolonged mean generation time (T) were signs of all disadvantageous effects associated with symbiotic reduction. Our findings confirmed the composition and richness of symbiotic bacteria in larva and adult of A. camelliae by an Illumina NovaSeq 6000 analysis and their influence on the development of the host by demographic research. Together, the results suggested that symbiotic bacteria play an important role in manipulating the biological development of their hosts, which might help us for developing new pest control agents and technologies for better management of A. camelliae.}, } @article {pmid37124042, year = {2023}, author = {Rodi, M and Gross, C and Sandri, TL and Berner, L and Marcet-Houben, M and Kocak, E and Pogoda, M and Casadei, N and Köhler, C and Kreidenweiss, A and Agnandji, ST and Gabaldón, T and Ossowski, S and Held, J}, title = {Whole genome analysis of two sympatric human Mansonella: Mansonella perstans and Mansonella sp "DEUX".}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1159814}, pmid = {37124042}, issn = {2235-2988}, mesh = {Animals ; Humans ; *Mansonella/genetics ; Phylogeny ; *Sympatry ; DNA, Ribosomal ; Amino Acid Sequence ; }, abstract = {INTRODUCTION: Mansonella species are filarial parasites that infect humans worldwide. Although these infections are common, knowledge of the pathology and diversity of the causative species is limited. Furthermore, the lack of sequencing data for Mansonella species, shows that their research is neglected. Apart from Mansonella perstans, a potential new species called Mansonella sp "DEUX" has been identified in Gabon, which is prevalent at high frequencies. We aimed to further determine if Mansonella sp "DEUX" is a genotype of M. perstans, or if these are two sympatric species.

METHODS: We screened individuals in the area of Fougamou, Gabon for Mansonella mono-infections and generated de novo assemblies from the respective samples. For evolutionary analysis, a phylogenetic tree was reconstructed, and the differences and divergence times are presented. In addition, mitogenomes were generated and phylogenies based on 12S rDNA and cox1 were created.

RESULTS: We successfully generated whole genomes for M. perstans and Mansonella sp "DEUX". Phylogenetic analysis based on annotated protein sequences, support the hypothesis of two distinct species. The inferred evolutionary analysis suggested, that M. perstans and Mansonella sp "DEUX" separated around 778,000 years ago. Analysis based on mitochondrial marker genes support our hypothesis of two sympatric human Mansonella species.

DISCUSSION: The results presented indicate that Mansonella sp "DEUX" is a new Mansonella species. These findings reflect the neglect of this research topic. And the availability of whole genome data will allow further investigations of these species.}, } @article {pmid37121168, year = {2023}, author = {Biney, C and Graham, GE and Asiedu, E and Sakyi, SA and Kwarteng, A}, title = {Wolbachia Ferrochelatase as a potential drug target against filarial infections.}, journal = {Journal of molecular graphics & modelling}, volume = {122}, number = {}, pages = {108490}, doi = {10.1016/j.jmgm.2023.108490}, pmid = {37121168}, issn = {1873-4243}, mesh = {Animals ; *Wolbachia/metabolism ; Ferrochelatase/metabolism/therapeutic use ; *Filariasis/drug therapy/parasitology ; *Brugia malayi ; Heme/metabolism ; }, abstract = {Filarial infections are among the world's most disturbing diseases caused by 3 major parasitic worms; Onchocerca volvulus, Wuchereria bancrofti, and Brugia malayi, affecting more than 500 million people worldwide. Currently used drugs for mass drug administration (MDA) have been met with several challenges including the development of complications in individuals with filaria co-infections and parasitic drug resistance. The filarial endosymbiont, Wolbachia, has emerged as an attractive therapeutic target for filariasis elimination, due to the dependence of the filaria on this endosymbiont for survival. Here, we target an important enzyme in the Wolbachia heme biosynthetic pathway (ferrochelatase), using high-throughput virtual screening and molecular dynamics with MM-PBSA calculations. We identified four drug candidates; Nilotinib, Ledipasvir, 3-benzhydryloxy-8-methyl-8-azabicyclo[3.2.1]octane, and 2-(4-Amino-piperidin-1-yl)-ethanol as potential small molecules inhibitors as they could compete with the enzyme's natural substrate (Protoporphyrin IX) for active pocket binding. This prevents the worm from receiving the heme molecule from Wolbachia for their growth and survival, resulting in their death. This study which involved targeting enzymes in biosynthetic pathways of the parasitic worms' endosymbiont (Wolbachia), has proven to be an alternative therapeutic option leading to the discovery of new drugs, which will help facilitate the elimination of parasitic infections.}, } @article {pmid37117399, year = {2023}, author = {Ghousein, A and Tutagata, J and Schrieke, H and Etienne, M and Chaumeau, V and Boyer, S and Pages, N and Roiz, D and Eren, AM and Cambray, G and Reveillaud, J}, title = {pWCP is a widely distributed and highly conserved Wolbachia plasmid in Culex pipiens and Culex quinquefasciatus mosquitoes worldwide.}, journal = {ISME communications}, volume = {3}, number = {1}, pages = {40}, pmid = {37117399}, issn = {2730-6151}, support = {948135//EC | EC Seventh Framework Programm | FP7 Ideas: European Research Council (FP7-IDEAS-ERC - Specific Programme: "Ideas" Implementing the Seventh Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (2007 to 2013))/ ; 220211//Wellcome Trust (Wellcome)/ ; }, abstract = {Mosquitoes represent the most important pathogen vectors and are responsible for the spread of a wide variety of poorly treatable diseases. Wolbachia are obligate intracellular bacteria that are widely distributed among arthropods and collectively represents one of the most promising solutions for vector control. In particular, Wolbachia has been shown to limit the transmission of pathogens, and to dramatically affect the reproductive behavior of their host through its phage WO. While much research has focused on deciphering and exploring the biocontrol applications of these WO-related phenotypes, the extent and potential impact of the Wolbachia mobilome remain poorly appreciated. Notably, several Wolbachia plasmids, carrying WO-like genes and Insertion Sequences (IS), thus possibly interrelated to other genetic units of the endosymbiont, have been recently discovered. Here we investigated the diversity and biogeography of the first described plasmid of Wolbachia in Culex pipiens (pWCP) in several islands and continental countries around the world-including Cambodia, Guadeloupe, Martinique, Thailand, and Mexico-together with mosquito strains from colonies that evolved for 2 to 30 years in the laboratory. We used PCR and qPCR to determine the presence and copy number of pWCP in individual mosquitoes, and highly accurate Sanger sequencing to evaluate potential variations. Together with earlier observation, our results show that pWCP is omnipresent and strikingly conserved among Wolbachia populations within mosquitoes from distant geographies and environmental conditions. These data suggest a critical role for the plasmid in Wolbachia ecology and evolution, and the potential of a great tool for further genetic dissection and possible manipulation of this endosymbiont.}, } @article {pmid37117271, year = {2023}, author = {Řezáč, M and Řezáčová, V and Gloríková, N and Némethová, E and Heneberg, P}, title = {Food provisioning to Pardosa spiders decreases the levels of tissue-resident endosymbiotic bacteria.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {6943}, pmid = {37117271}, issn = {2045-2322}, mesh = {Animals ; *Spiders/microbiology ; Symbiosis ; *Coxiellaceae ; Host Specificity ; Drosophila ; *Rickettsia ; }, abstract = {The diversity, host specificity, and physiological effects of endosymbiotic bacteria in spiders (Araneae) are poorly characterized. We used 16S rDNA sequencing to evaluate endosymbionts in the cephalothorax and legs of a wolf spider Pardosa agrestis. We tested the effects of feeding once or twice daily with fruit flies, aphids, or starved and compared them to those of syntopically occurring Pardosa palustris. The feeding increased traveled distance up to five times in some of the groups provisioned with food relative to the starved control. The Shannon diversity t-test revealed significant differences between these component communities of the two spider species. The increased frequency of feeding with fruit flies, but not aphids, increased the dominance and decreased the alpha diversity of OTUs. The obligate or facultative endosymbionts were present in all analyzed spider individuals and were represented mostly by Rickettsiella, Rhabdochlamydia, Spiroplasma, and the facultative intracellular parasite Legionella. Vertically transmitted endosymbionts were less common, represented by Wolbachia pipientis and Rickettsia sp. H820. The relative abundance of Mycoplasma spp. was negatively correlated with provisioned or killed aphids. In conclusion, the tissues of Pardosa spiders host tremendously diverse assemblages of bacteria, including obligate or facultative endosymbionts, with yet unknown phenotypic effects.}, } @article {pmid37112932, year = {2023}, author = {Petersen, MT and Couto-Lima, D and Garcia, GA and Pavan, MG and David, MR and Maciel-de-Freitas, R}, title = {Dengue Exposure and Wolbachia wMel Strain Affects the Fertility of Quiescent Eggs of Aedes aegypti.}, journal = {Viruses}, volume = {15}, number = {4}, pages = {}, pmid = {37112932}, issn = {1999-4915}, mesh = {Humans ; Animals ; Female ; *Dengue Virus ; *Aedes ; *Wolbachia ; Fertility ; *Dengue ; }, abstract = {(1) Background: The deployment of the bacterium Wolbachia to reduce arbovirus transmission is ongoing in several countries worldwide. When Wolbachia-carrying Aedes aegypti are released and established in the field, females may feed on dengue-infected hosts. The effects of simultaneous exposure on life-history traits of Ae. aegypti to Wolbachia wMel strain and dengue-1 virus DENV-1 remain unclear. (2) Methods: We monitored 4 groups (mosquitoes with either DENV-1 or Wolbachia, coinfected with DENV-1 and Wolbachia, as well as negative controls) to estimate Ae. aegypti survival, oviposition success, fecundity, collapsing and fertility of quiescent eggs for 12 weeks. (3) Results: Neither DENV-1 nor Wolbachia had a significant impact on mosquito survival nor on mosquito fecundity, although the last parameter showed a tendency to decrease with ageing. There was a significant decrease in oviposition success in individuals carrying Wolbachia. Wolbachia infection and storage time significantly increased egg collapse parameter on the egg viability assay, while DENV-1 had a slight protective effect on the first four weeks of storage. (4) Conclusions: Despite limitations, our results contribute to better understanding of the tripartite interaction of virus, bacteria and mosquito that may take place in field conditions and aid in guaranteeing the Wolbachia strategy success.}, } @article {pmid37105498, year = {2023}, author = {Gong, JT and Li, TP and Wang, MK and Hong, XY}, title = {Wolbachia-based strategies for control of agricultural pests.}, journal = {Current opinion in insect science}, volume = {57}, number = {}, pages = {101039}, doi = {10.1016/j.cois.2023.101039}, pmid = {37105498}, issn = {2214-5753}, mesh = {Humans ; Animals ; *Aedes ; *Wolbachia ; Population Dynamics ; }, abstract = {Wolbachia-based incompatible insect technique (IIT) and pathogen blocking technique (PBT) have been shown to be effective at protecting humans from mosquito-borne diseases in the past decades. Population suppression based on IIT and population replacement based on PBT have become major field application strategies that have continuously been improved by the translational research on Wolbachia-transinfected mosquitoes. Similarly, Wolbachia-based approaches have been proposed for the protection of plants from agricultural pests and their associated diseases. However, a bottleneck in Wolbachia-based strategies for the control of agricultural pests is the need for methods to establish Wolbachia-transinfected insect lines. As a first step in this direction, we compare field control strategies for mosquitos with the potential strategies for agricultural pests based on Wolbachia. Our results show that there is a critical need for establishing productive insect lines and accumulating field test data.}, } @article {pmid37103495, year = {2023}, author = {Towett-Kirui, S and Morrow, JL and Close, S and Royer, JE and Riegler, M}, title = {Bacterial Communities Are Less Diverse in a Strepsipteran Endoparasitoid than in Its Fruit Fly Hosts and Dominated by Wolbachia.}, journal = {Microbial ecology}, volume = {86}, number = {3}, pages = {2120-2132}, pmid = {37103495}, issn = {1432-184X}, support = {IC150100026//Australian Research Council (ARC) Industrial Transformation Training Centre (ITTC) Fruit Fly Biosecurity Innovation/ ; }, mesh = {*Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Bacteria/genetics ; *Microbiota ; High-Throughput Nucleotide Sequencing ; }, abstract = {Microbiomes play vital roles in insect fitness and health and can be influenced by interactions between insects and their parasites. Many studies investigate the microbiome of free-living insects, whereas microbiomes of endoparasitoids and their interactions with parasitised insects are less explored. Due to their development in the constrained environment within a host, endoparasitoids are expected to have less diverse yet distinct microbiomes. We used high-throughput 16S rRNA gene amplicon sequencing to characterise the bacterial communities of Dipterophagus daci (Strepsiptera) and seven of its tephritid fruit fly host species. Bacterial communities of D. daci were less diverse and contained fewer taxa relative to the bacterial communities of the tephritid hosts. The strepsipteran's microbiome was dominated by Pseudomonadota (formerly Proteobacteria) (> 96%), mainly attributed to the presence of Wolbachia, with few other bacterial community members, indicative of an overall less diverse microbiome in D. daci. In contrast, a dominance of Wolbachia was not found in flies parasitised by early stages of D. daci nor unparasitised flies. Yet, early stages of D. daci parasitisation resulted in structural changes in the bacterial communities of parasitised flies. Furthermore, parasitisation with early stages of D. daci with Wolbachia was associated with a change in the relative abundance of some bacterial taxa relative to parasitisation with early stages of D. daci lacking Wolbachia. Our study is a first comprehensive characterisation of bacterial communities in a Strepsiptera species together with the more diverse bacterial communities of its hosts and reveals effects of concealed stages of parasitisation on host bacterial communities.}, } @article {pmid37103216, year = {2023}, author = {Lv, N and Peng, J and He, ZQ and Wen, Q and Su, ZQ and Ali, S and Liu, CZ and Qiu, BL}, title = {The Dynamic Distribution of Wolbachia and Rickettsia in AsiaII1 Bemisia tabaci.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103216}, issn = {2075-4450}, support = {32060250//the National Science Foundation of China/ ; }, abstract = {Wolbachia and Rickettsia are bacterial endosymbionts that can induce a number of reproductive abnormalities in their arthropod hosts. We screened and established the co-infection of Wolbachia and Rickettsia in Bemisia tabaci and compared the spatial and temporal distribution of Wolbachia and Rickettsia in eggs (3-120 h after spawning), nymphs, and adults of B. tabaci by qPCR quantification and fluorescent in situ hybridization (FISH). The results show that the titer of Wolbachia and Rickettsia in the 3-120 h old eggs showed a "w" patterned fluctuation, while the titers of Wolbachia and Rickettsia had a "descending-ascending descending-ascending" change process. The titers of Rickettsia and Wolbachia nymphal and the adult life stages of Asia II1 B. tabaci generally increased with the development of whiteflies. However, the location of Wolbachia and Rickettsia in the egg changed from egg stalk to egg base, and then from egg base to egg posterior, and finally back to the middle of the egg. These results will provide basic information on the quantity and localization of Wolbachia and Rickettsia within different life stages of B. tabaci. These findings help to understand the dynamics of the vertical transmission of symbiotic bacteria.}, } @article {pmid37103189, year = {2023}, author = {Cheong, YL and Nazni, WA and Lee, HL and NoorAfizah, A and MohdKhairuddin, IC and Kamarul, GMR and Nizam, NMN and Arif, MAK and NurZatilAqmar, ZM and Irwan, SM and Khadijah, K and Paid, YM and Topek, O and Hasnor, AH and AbuBakar, R and Singh Gill, B and Fadzilah, K and Tahir, A and Sinkins, SP and Hoffmann, AA}, title = {Spatial Distribution and Long-Term Persistence of Wolbachia-Infected Aedes aegypti in the Mentari Court, Malaysia.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103189}, issn = {2075-4450}, support = {Wellcome Trust Award 108508 and Wellcome Trust Award 108508/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Dengue is endemic in Malaysia, and vector control strategies are vital to reduce dengue transmission. The Wolbachia strain wAlbB carried by both sexes of Ae. aegypti was released in Mentari Court, a high-rise residential site, in October 2017 and stopped after 20 weeks. Wolbachia frequencies are still being monitored at multiple traps across this site, providing an opportunity to examine the spatiotemporal distribution of Wolbachia and mosquito density with respect to year, residential block, and floor, using spatial interpolation in ArcGIS, GLMs, and contingency analyses. In just 12 weeks, Wolbachia-infected mosquitoes were established right across the Mentari Court site with an overall infection frequency of >90%. To date, the Wolbachia frequency of Ae. aegypti has remained high in all areas across the site despite releases finishing four years ago. Nevertheless, the Wolbachia invaded more rapidly in some residential blocks than others, and also showed a relatively higher frequency on the eighth floor. The Ae. aegypti index tended to differ somewhat between residential blocks, whilst the Ae. albopictus index was relatively higher at the top and bottom floors of buildings. In Mentari Court, only a short release period was required to infiltrate Wolbachia completely and stably into the natural population. The results inform future releases in comparable sites in a dengue control programme.}, } @article {pmid37103172, year = {2023}, author = {Karpova, EK and Bobrovskikh, MA and Deryuzhenko, MA and Shishkina, OD and Gruntenko, NE}, title = {Wolbachia Effect on Drosophila melanogaster Lipid and Carbohydrate Metabolism.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103172}, issn = {2075-4450}, support = {19-04-00458//Russian Foundation for Basic Research/ ; }, abstract = {The effect of maternally inherited endosymbiotic bacteria Wolbachia on triglyceride and carbohydrate metabolism, starvation resistance and feeding behavior of Drosophila melanogaster females was studied. Eight D. melanogaster lines of the same nuclear background were investigated; one had no infection and served as the control, and seven others were infected with different Wolbachia strains pertaining to wMel and wMelCS groups of genotypes. Most of the infected lines had a higher overall lipid content and triglyceride level than the control line and their expression of the bmm gene regulating triglyceride catabolism was reduced. The glucose content was higher in the infected lines compared to that in the control, while their trehalose levels were similar. It was also found that the Wolbachia infection reduced the level of tps1 gene expression (coding for enzyme for trehalose synthesis from glucose) and had no effect on treh gene expression (coding for trehalose degradation enzyme). The infected lines exhibited lower appetite but higher survival under starvation compared to the control. The data obtained may indicate that Wolbachia foster their hosts' energy exchange through increasing its lipid storage and glucose content to ensure the host's competitive advantage over uninfected individuals. The scheme of carbohydrate and lipid metabolism regulation under Wolbachia's influence was suggested.}, } @article {pmid37103136, year = {2023}, author = {Heffernan, E and Markee, A and Truglio, MR and Barkdull, M and Steele Cabrera, S and Daniels, J}, title = {Population Genetic Structure of a Rare Butterfly in a Fragmented South Florida Ecosystem.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103136}, issn = {2075-4450}, support = {NA//Disney Conservation Fund/ ; }, abstract = {We investigated the genetic structure and diversity between populations of a rare butterfly, the Florida duskywing (Ephyriades brunnea floridensis E. Bell and W. Comstock, 1948) (Lepidoptera: Hesperiidae) across a network of South Florida pine rockland habitat fragments. Based on 81 individuals from seven populations and using multiple polymorphic microsatellite loci, our analyses support the presence of mainland Florida (peninsular) and Florida Keys (island) population groupings, with a moderate, asymmetrical gene flow connecting them, and the presence of private alleles providing unique identities to each. We additionally found that despite a prevalence in many Lepidoptera, the presence of Wolbachia was not identified in any of the samples screened. Our findings can be used to inform conservation and recovery decisions, including population monitoring, organism translocation, and priority areas for management, restoration or stepping-stone creation to help maintain the complex genetic structure of separate populations.}, } @article {pmid37098937, year = {2023}, author = {Arai, H and Takamatsu, T and Lin, SR and Mizutani, T and Omatsu, T and Katayama, Y and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Diverse Molecular Mechanisms Underlying Microbe-Inducing Male Killing in the Moth Homona magnanima.}, journal = {Applied and environmental microbiology}, volume = {89}, number = {5}, pages = {e0209522}, pmid = {37098937}, issn = {1098-5336}, mesh = {Animals ; Female ; Male ; *Moths ; Symbiosis ; Larva/microbiology ; Reproduction ; Apoptosis ; *Wolbachia/genetics ; *Spiroplasma/genetics ; }, abstract = {Male killing (MK) is a type of reproductive manipulation induced by microbes, where sons of infected mothers are killed during development. MK is a strategy that enhances the fitness of the microbes, and the underlying mechanisms and the process of their evolution have attracted substantial attention. Homona magnanima, a moth, harbors two embryonic MK bacteria, namely, Wolbachia (Alphaproteobacteria) and Spiroplasma (Mollicutes), and a larval MK virus, Osugoroshi virus (OGV; Partitiviridae). However, whether the three distantly related male killers employ similar or different mechanisms to accomplish MK remains unknown. Here, we clarified the differential effects of the three male killers on the sex-determination cascades and development of H. magnanima males. Reverse transcription-PCR demonstrated that Wolbachia and Spiroplasma, but not OGVs, disrupted the sex-determination cascade of males by inducing female-type splice variants of doublesex (dsx), a downstream regulator of the sex-determining gene cascade. We also found that MK microbes altered host transcriptomes in different manners; Wolbachia impaired the host dosage compensation system, whereas Spiroplasma and OGVs did not. Moreover, Wolbachia and Spiroplasma, but not OGVs, triggered abnormal apoptosis in male embryos. These findings suggest that distantly related microbes employ distinct machineries to kill males of the identical host species, which would be the outcome of the convergent evolution. IMPORTANCE Many microbes induce male killing (MK) in various insect species. However, it is not well understood whether microbes adopt similar or different MK mechanisms. This gap in our knowledge is partly because different insect models have been examined for each MK microbe. Here, we compared three taxonomically distinct male killers (i.e., Wolbachia, Spiroplasma, and a partiti-like virus) that infect the same host. We provided evidence that microbes can cause MK through distinct mechanisms that differ in the expression of genes involved in sex determination, dosage compensation, and apoptosis. These results imply independent evolutionary scenarios for the acquisition of their MK ability.}, } @article {pmid37098535, year = {2023}, author = {da Moura, AJF and Valadas, V and Da Veiga Leal, S and Montalvo Sabino, E and Sousa, CA and Pinto, J}, title = {Screening of natural Wolbachia infection in mosquitoes (Diptera: Culicidae) from the Cape Verde islands.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {142}, pmid = {37098535}, issn = {1756-3305}, support = {PhD fellowship//Camões I.P/ ; PTDC/BIA-OUT/29477/2017//Fundação para a Ciência e a Tecnologia/ ; }, mesh = {Animals ; *Culicidae/genetics ; *Wolbachia/genetics ; Multilocus Sequence Typing ; Cabo Verde ; Mosquito Vectors/microbiology ; *Culex/genetics ; *Aedes/genetics ; }, abstract = {BACKGROUND: Wolbachia pipientis is an endosymbiont bacterium that induces cytoplasmic incompatibility and inhibits arboviral replication in mosquitoes. This study aimed to assess Wolbachia prevalence and genetic diversity in different mosquito species from Cape Verde.

METHODS: Mosquitoes were collected on six islands of Cape Verde and identified to species using morphological keys and PCR-based assays. Wolbachia was detected by amplifying a fragment of the surface protein gene (wsp). Multilocus sequence typing (MLST) was performed with five housekeeping genes (coxA, gatB, ftsZ, hcpA, and fbpA) and the wsp hypervariable region (HVR) for strain identification. Identification of wPip groups (wPip-I to wPip-V) was performed using PCR-restriction fragment length polymorphism (RFLP) assay on the ankyrin domain gene pk1.

RESULTS: Nine mosquito species were collected, including the major vectors Aedes aegypti, Anopheles arabiensis, Culex pipiens sensu stricto, and Culex quinquefasciatus. Wolbachia was only detected in Cx. pipiens s.s. (100% prevalence), Cx. quinquefasciatus (98.3%), Cx. pipiens/quinquefasciatus hybrids (100%), and Culex tigripes (100%). Based on the results of MLST and wsp hypervariable region typing, Wolbachia from the Cx. pipiens complex was assigned to sequence type 9, wPip clade, and supergroup B. PCR/RFLP analysis revealed three wPip groups in Cape Verde, namely wPip-II, wPip-III, and wPip-IV. wPip-IV was the most prevalent, while wPip-II and wPip-III were found only on Maio and Fogo islands. Wolbachia detected in Cx. tigripes belongs to supergroup B, with no attributed MLST profile, indicating a new strain of Wolbachia in this mosquito species.

CONCLUSIONS: A high prevalence and diversity of Wolbachia was found in species from the Cx. pipiens complex. This diversity may be related to the mosquito's colonization history on the Cape Verde islands. To the best of our knowledge, this is the first study to detect Wolbachia in Cx. tigripes, which may provide an additional opportunity for biocontrol initiatives.}, } @article {pmid37094148, year = {2023}, author = {Gu, X and Ross, PA and Gill, A and Yang, Q and Ansermin, E and Sharma, S and Soleimannejad, S and Sharma, K and Callahan, A and Brown, C and Umina, PA and Kristensen, TN and Hoffmann, AA}, title = {A rapidly spreading deleterious aphid endosymbiont that uses horizontal as well as vertical transmission.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {120}, number = {18}, pages = {e2217278120}, pmid = {37094148}, issn = {1091-6490}, mesh = {Animals ; *Aphids/microbiology ; *Coxiellaceae/genetics ; Bacteria ; Phenotype ; Reproduction ; Symbiosis ; }, abstract = {Endosymbiotic bacteria that live inside the cells of insects are typically only transmitted maternally and can spread by increasing host fitness and/or modifying reproduction in sexual hosts. Transinfections of Wolbachia endosymbionts are now being used to introduce useful phenotypes into sexual host populations, but there has been limited progress on applications using other endosymbionts and in asexual populations. Here, we develop a unique pathway to application in aphids by transferring the endosymbiont Rickettsiella viridis to the major crop pest Myzus persicae. Rickettsiella infection greatly reduced aphid fecundity, decreased heat tolerance, and modified aphid body color, from light to dark green. Despite inducing host fitness costs, Rickettsiella spread rapidly through caged aphid populations via plant-mediated horizontal transmission. The phenotypic effects of Rickettsiella were sensitive to temperature, with spread only occurring at 19 °C and not 25 °C. Body color modification was also lost at high temperatures despite Rickettsiella maintaining a high density. Rickettsiella shows the potential to spread through natural M. persicae populations by horizontal transmission and subsequent vertical transmission. Establishment of Rickettsiella in natural populations could reduce crop damage by modifying population age structure, reducing population growth and providing context-dependent effects on host fitness. Our results highlight the importance of plant-mediated horizontal transmission and interactions with temperature as drivers of endosymbiont spread in asexual insect populations.}, } @article {pmid37079598, year = {2023}, author = {Choubdar, N and Karimian, F and Koosha, M and Nejati, J and Shabani Kordshouli, R and Azarm, A and Oshaghi, MA}, title = {Wolbachia infection in native populations of Blattella germanica and Periplaneta americana.}, journal = {PloS one}, volume = {18}, number = {4}, pages = {e0284704}, pmid = {37079598}, issn = {1932-6203}, mesh = {Animals ; *Periplaneta/microbiology ; *Blattellidae/genetics/microbiology ; *Wolbachia/genetics ; Phylogeny ; *Cockroaches/microbiology ; Allergens ; }, abstract = {Cockroaches are significant pests worldwide, being important in medical, veterinary, and public health fields. Control of cockroaches is difficult because they have robust reproductive ability and high adaptability and are resistant to many insecticides. Wolbachia is an endosymbiont bacterium that infects the reproductive organs of approximately 70% of insect species and has become a promising biological agent for controlling insect pests. However, limited data on the presence or strain typing of Wolbachia in cockroaches are available. PCR amplification and sequencing of the wsp and gltA genes were used to study the presence, prevalence and molecular typing of Wolbachia in two main cockroach species, Blattella germanica (German cockroach) and Periplaneta americana (American cockroach), from different geographical locations of Iran. The Wolbachia endosymbiont was found only in 20.6% of German cockroaches while it was absent in American cockroach samples. Blast search and phylogenetic analysis revealed that the Wolbachia strain found in the German cockroach belongs to Wolbachia supergroup F. Further studies should investigate the symbiotic role of Wolbachia in cockroaches and determine whether lack of Wolbachia infection may increase this insect's ability to tolerate or acquire various pathogens. Results of our study provide a foundation for continued work on interactions between cockroaches, bacterial endosymbionts, and pathogens.}, } @article {pmid37075471, year = {2023}, author = {Che Lah, EF and Ahamad, M and Dmitry, A and Md-Zain, BM and Yaakop, S}, title = {Metagenomic profile of the bacterial communities associated with Ixodes granulatus (Acari: Ixodidae): a potential vector of tick-borne diseases.}, journal = {Journal of medical entomology}, volume = {60}, number = {4}, pages = {753-768}, doi = {10.1093/jme/tjad044}, pmid = {37075471}, issn = {1938-2928}, mesh = {Humans ; Female ; Animals ; *Ixodes/microbiology ; *Ixodidae/microbiology ; Rickettsiales ; Ehrlichia ; *Rickettsia/genetics ; *Borrelia/genetics ; *Tick-Borne Diseases/microbiology ; }, abstract = {Ixodes granulatus Supino, 1897 (Acari: Ixodida) is one of Malaysia's most common hard ticks and is a potential vector for tick-borne diseases (TBDs). Despite its great public health importance, research on I. granulatus microbial communities remains largely unexplored. Therefore, this study aimed to investigate the bacterial communities of on-host I. granulatus collected from three different recreational areas on the East Coast of Peninsular Malaysia using high throughput Next Generation Sequencing (NGS). A total of 9 females on-host I. granulatus were subjected to metabarcoding analysis targeting V3-V4 regions of 16S ribosomal RNA (rRNA) using the Illumina MiSeq platform. This study identified 15 bacterial phyla corresponding to 19 classes, 54 orders, and 90 families from 435 amplicon sequence variants (ASVs), revealing a diverse bacterial community profile. Together with 130 genera assigned, local I. granulatus harbored 4 genera of pathogens, i.e., Rickettsia da Rocha Lima, 1916 (Rickettsiales: Rickettsiaceae) (58.6%), Borrelia Swellengrebel 1907 (Spirochaetales: Borreliaceae) (31.6%), Borreliella Adeolu and Gupta 2015 (Spirochaetales: Borreliaceae) (0.6%), and Ehrlichia Cowdria Moshkovski 1947 (Rickettsiales: Ehrlichiaceae) (39.9%). Some endosymbiont bacteria, such as Coxiella (Philip, 1943) (Legionellales: Coxiellaceae), Wolbachia Hertig 1936 (Rickettsiales: Ehrlichiaceae), and Rickettsiella Philip, 1956 (Legionellales: Coxiellaceae), were also detected at very low abundance. Interestingly, this study reported the co-infection of Borrelia and Ehrlichia for the first time, instilling potential health concerns in the context of co-transmission to humans, especially in areas with a high population of I. granulatus. This study successfully characterized the tick microbiome and provided the first baseline data of I. granulatus bacterial communities in Malaysia. These results support the need for way-forward research on tick-associated bacteria using NGS, focusing on medically important species toward TBD prevention.}, } @article {pmid37073332, year = {2023}, author = {Ye, Z and Bishop, T and Wang, Y and Shahriari, R and Lynch, M}, title = {Evolution of sex determination in crustaceans.}, journal = {Marine life science & technology}, volume = {5}, number = {1}, pages = {1-11}, pmid = {37073332}, issn = {2662-1746}, abstract = {Sex determination (SD) involves mechanisms that determine whether an individual will develop into a male, female, or in rare cases, hermaphrodite. Crustaceans harbor extremely diverse SD systems, including hermaphroditism, environmental sex determination (ESD), genetic sex determination (GSD), and cytoplasmic sex determination (e.g., Wolbachia controlled SD systems). Such diversity lays the groundwork for researching the evolution of SD in crustaceans, i.e., transitions among different SD systems. However, most previous research has focused on understanding the mechanism of SD within a single lineage or species, overlooking the transition across different SD systems. To help bridge this gap, we summarize the understanding of SD in various clades of crustaceans, and discuss how different SD systems might evolve from one another. Furthermore, we review the genetic basis for transitions between different SD systems (i.e., Dmrt genes) and propose the microcrustacean Daphnia (clade Branchiopoda) as a model to study the transition from ESD to GSD.}, } @article {pmid37060070, year = {2023}, author = {Li, Y and Sun, Y and Zou, J and Zhong, D and Liu, R and Zhu, C and Li, W and Zhou, Y and Cui, L and Zhou, G and Lu, G and Li, T}, title = {Characterizing the Wolbachia infection in field-collected Culicidae mosquitoes from Hainan Province, China.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {128}, pmid = {37060070}, issn = {1756-3305}, support = {820RC653//Hainan Provincial Natural Science Foundation/ ; 82060379//National Natural Science Foundation of China/ ; 2022NHCTDCKFKT31002//Open Foundation of NHC Key Laboratory of Tropical Disease Control, Hainan Medical University/ ; U19 AI089672/AI/NIAID NIH HHS/United States ; ZDKJ202003//Major Science and Technology Program of Hainan Province/ ; YSPTZX202004//Research project of Hainan academician innovation platform/ ; XRC220012//Talent Introduction Fund of Hainan Medical University/ ; }, mesh = {Animals ; Humans ; *Culicidae ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; Mosquito Vectors/genetics ; *Aedes/genetics ; *Culex/genetics ; China/epidemiology ; }, abstract = {BACKGROUND: Mosquitoes are vectors of many pathogens, such as malaria, dengue virus, yellow fever virus, filaria and Japanese encephalitis virus. Wolbachia are capable of inducing a wide range of reproductive abnormalities in their hosts, such as cytoplasmic incompatibility. Wolbachia has been proposed as a tool to modify mosquitoes that are resistant to pathogen infection as an alternative vector control strategy. This study aimed to determine natural Wolbachia infections in different mosquito species across Hainan Province, China.

METHODS: Adult mosquitoes were collected using light traps, human landing catches and aspirators in five areas in Hainan Province from May 2020 to November 2021. Species were identified based on morphological characteristics, species-specific PCR and DNA barcoding of cox1 assays. Molecular classification of species and phylogenetic analyses of Wolbachia infections were conducted based on the sequences from PCR products of cox1, wsp, 16S rRNA and FtsZ gene segments.

RESULTS: A total of 413 female adult mosquitoes representing 15 species were identified molecularly and analyzed. Four mosquito species (Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Culex gelidus) were positive for Wolbachia infection. The overall Wolbachia infection rate for all mosquitoes tested in this study was 36.1% but varied among species. Wolbachia types A, B and mixed infections of A × B were detected in Ae. albopictus mosquitoes. A total of five wsp haplotypes, six FtsZ haplotypes and six 16S rRNA haplotypes were detected from Wolbachia infections. Phylogenetic tree analysis of wsp sequences classified them into three groups (type A, B and C) of Wolbachia strains compared to two groups each for FtsZ and 16S rRNA sequences. A novel type C Wolbachia strain was detected in Cx. gelidus by both single locus wsp gene and the combination of three genes.

CONCLUSION: Our study revealed the prevalence and distribution of Wolbachia in mosquitoes from Hainan Province, China. Knowledge of the prevalence and diversity of Wolbachia strains in local mosquito populations will provide part of the baseline information required for current and future Wolbachia-based vector control approaches to be conducted in Hainan Province.}, } @article {pmid37055642, year = {2023}, author = {Lozano-Sardaneta, YN and Marina, CF and Torres-Monzón, JA and Sánchez-Cordero, V and Becker, I}, title = {Molecular detection of Wolbachia and Bartonella as part of the microbiome of phlebotomine sand flies from Chiapas, Mexico.}, journal = {Parasitology research}, volume = {122}, number = {6}, pages = {1293-1301}, pmid = {37055642}, issn = {1432-1955}, support = {6682//Consejo Nacional de Ciencia y Tecnología/ ; CRISP-CONACYT-FOSSIS-69530//Consejo Nacional de Ciencia y Tecnología/ ; IG201221//Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica (PAPIIT)/ ; }, mesh = {Animals ; *Psychodidae/parasitology ; *Wolbachia/genetics ; *Bartonella/genetics ; Mexico ; Phylogeny ; Retrospective Studies ; *Phlebotomus ; *Leishmania ; *Microbiota ; Bacteria ; }, abstract = {Phlebotomine sand flies are dipterans of relevance due to their role as vectors of several pathogens worldwide. Bacteria in the gut of sand flies possibly affect their vectorial capacity and competence to transmit parasites. A retrospective study was performed in sand fly specimens that had previously been collected in four localities of the state of Chiapas during the period 2009-2011 to detect Wolbachia and Bartonella and their possible coinfection with Leishmania. For the molecular detection of bacteria, we used primers and conditions that had previously been reported. A total of 531 sand fly specimens of 10 species were analyzed. Four Wolbachia strains were detected in five sand fly species, showing a prevalence of 8.6%. All the Wolbachia strains had previously been reported in other taxa. In one sand fly species, we also detected a new lineage of Bartonella evidenced by a phylogenetic analysis. No sand fly specimens showed coinfections of these bacteria and Leishmania. The bacteria found in the phlebotomine sand flies are possibly transmitted by plant-mediated horizontal transmission and during blood meal feeding.}, } @article {pmid37052365, year = {2023}, author = {Xu, J and Tan, JB and Li, YD and Xu, YH and Tang, A and Zhou, HK and Shi, PQ}, title = {Diversity and dynamics of endosymbionts in a single population of sweet potato weevil, Cylas formicarius (Coleoptera: Brentidae): a preliminary study.}, journal = {Journal of insect science (Online)}, volume = {23}, number = {2}, pages = {}, pmid = {37052365}, issn = {1536-2442}, mesh = {Animals ; *Weevils ; *Coleoptera ; *Ipomoea batatas ; Reproduction ; }, abstract = {Endosymbionts live symbiotically with insect hosts and play important roles in the evolution, growth, development, reproduction, and environmental fitness of hosts. Weevils are one of the most abundant insect groups that can be infected by various endosymbionts, such as Sodalis, Nardonella, and Wolbachia. The sweet potato weevil, Cylas formicarius (Coleoptera: Brentidae), is a notorious pest in sweet potato (Ipomoea batatas L.) cultivation. Currently, little is known about the presence of endosymbionts in C. formicarius. Herein, we assessed the endosymbiont load of a single geographic population of C. formicarius. The results showed that Nardonella and Rickettsia could infect C. formicarius at different rates, which also varied according to the developmental stages of C. formicarius. The relative titer of Nardonella was significantly related to C. formicarius developmental stages. The Nardonella-infecting sweet potato weevils were most closely related to the Nardonella in Sphenophorus levis (Coleoptera, Curculionidae). The Rickettsia be identified in bellii group. These results preliminarily revealed the endosymbionts in C. formicarius and helped to explore the diversity of endosymbionts in weevils and uncover the physiological roles of endosymbionts in weevils.}, } @article {pmid37043605, year = {2023}, author = {Dobson, KL and Blore, K and Henke, JA and Hung, KY and Morgan, T and Posey, T and Sun, S and Sypes, O and Tremblay, NP and Dobson, SL}, title = {Satellite Rearing of Aedes Mosquito Eggs: Synchronized Empirical Test of a Novel Mass Rearing Model.}, journal = {Journal of the American Mosquito Control Association}, volume = {39}, number = {1}, pages = {12-17}, doi = {10.2987/22-7104}, pmid = {37043605}, issn = {1943-6270}, mesh = {Animals ; Male ; *Aedes ; Humidity ; Mosquito Control/methods ; }, abstract = {Mosquito suppression strategies based on "rear and release" of male mosquitoes are attracting renewed interest from governments, municipalities, and private businesses. These include irradiation-based sterile insect technique, Wolbachia-based technologies, and genetic modification. Each of these approaches requires the mass rearing and release of adult male mosquitoes, which typically is accomplished via a rearing facility near the release site. Although some release programs have relied on centralized rearing and shipment of adult males, adult male mosquitoes are relatively fragile, and their fitness can be diminished by temperature fluctuations, humidity, nutritional deficiencies, and other stresses that occur during shipment. Furthermore, expensive, expedited shipment is typically used to maximize the amount of adult lifetime in the field following the release. In contrast, Aedes aegypti and Ae. albopictus eggs can be desiccated and stored for long periods. They are small, and many millions of eggs can be shipped without specialized environmental conditions and using less expensive means. Here we examine a model in which mosquito eggs are centrally produced and then mailed to satellite rearing facilities. As a control, a replicate set of eggs was reared at the factory of origin. At each of the rearing sites, cloud-based software was used to track and compare rearing at the different locations. The results demonstrate similar rearing outcomes (i.e., egg hatch, immature development, and number of adult males) at each of the different sites for both species. We discuss the outcome in relation to downstream applications and potential future studies.}, } @article {pmid37043515, year = {2023}, author = {Novelo, M and Dutra, HL and Metz, HC and Jones, MJ and Sigle, LT and Frentiu, FD and Allen, SL and Chenoweth, SF and McGraw, EA}, title = {Dengue and chikungunya virus loads in the mosquito Aedes aegypti are determined by distinct genetic architectures.}, journal = {PLoS pathogens}, volume = {19}, number = {4}, pages = {e1011307}, pmid = {37043515}, issn = {1553-7374}, mesh = {Animals ; *Chikungunya virus/physiology ; *Aedes ; Mosquito Vectors ; *Chikungunya Fever ; *Dengue ; }, abstract = {Aedes aegypti is the primary vector of the arboviruses dengue (DENV) and chikungunya (CHIKV). These viruses exhibit key differences in their vector interactions, the latter moving more quicky through the mosquito and triggering fewer standard antiviral pathways. As the global footprint of CHIKV continues to expand, we seek to better understand the mosquito's natural response to CHIKV-both to compare it to DENV:vector coevolutionary history and to identify potential targets in the mosquito for genetic modification. We used a modified full-sibling design to estimate the contribution of mosquito genetic variation to viral loads of both DENV and CHIKV. Heritabilities were significant, but higher for DENV (40%) than CHIKV (18%). Interestingly, there was no genetic correlation between DENV and CHIKV loads between siblings. These data suggest Ae. aegypti mosquitoes respond to the two viruses using distinct genetic mechanisms. We also examined genome-wide patterns of gene expression between High and Low CHIKV families representing the phenotypic extremes of viral load. Using RNAseq, we identified only two loci that consistently differentiated High and Low families: a long non-coding RNA that has been identified in mosquito screens post-infection and a distant member of a family of Salivary Gland Specific (SGS) genes. Interestingly, the latter gene is also associated with horizontal gene transfer between mosquitoes and the endosymbiotic bacterium Wolbachia. This work is the first to link the SGS gene to a mosquito phenotype. Understanding the molecular details of how this gene contributes to viral control in mosquitoes may, therefore, also shed light on its role in Wolbachia.}, } @article {pmid37035661, year = {2023}, author = {Michalik, A and Franco, DC and Deng, J and Szklarzewicz, T and Stroiński, A and Kobiałka, M and Łukasik, P}, title = {Variable organization of symbiont-containing tissue across planthoppers hosting different heritable endosymbionts.}, journal = {Frontiers in physiology}, volume = {14}, number = {}, pages = {1135346}, pmid = {37035661}, issn = {1664-042X}, abstract = {Sap-feeding hemipteran insects live in associations with diverse heritable symbiotic microorganisms (bacteria and fungi) that provide essential nutrients deficient in their hosts' diets. These symbionts typically reside in highly specialized organs called bacteriomes (with bacterial symbionts) or mycetomes (with fungal symbionts). The organization of these organs varies between insect clades that are ancestrally associated with different microbes. As these symbioses evolve and additional microorganisms complement or replace the ancient associates, the organization of the symbiont-containing tissue becomes even more variable. Planthoppers (Hemiptera: Fulgoromorpha) are ancestrally associated with bacterial symbionts Sulcia and Vidania, but in many of the planthopper lineages, these symbionts are now accompanied or have been replaced by other heritable bacteria (e.g., Sodalis, Arsenophonus, Purcelliella) or fungi. We know the identity of many of these microbes, but the symbiont distribution within the host tissues and the bacteriome organization have not been systematically studied using modern microscopy techniques. Here, we combine light, fluorescence, and transmission electron microscopy with phylogenomic data to compare symbiont tissue distributions and the bacteriome organization across planthoppers representing 15 families. We identify and describe seven primary types of symbiont localization and seven types of the organization of the bacteriome. We show that Sulcia and Vidania, when present, usually occupy distinct bacteriomes distributed within the body cavity. The more recently acquired gammaproteobacterial and fungal symbionts generally occupy separate groups of cells organized into distinct bacteriomes or mycetomes, distinct from those with Sulcia and Vidania. They can also be localized in the cytoplasm of fat body cells. Alphaproteobacterial symbionts colonize a wider range of host body habitats: Asaia-like symbionts often colonize the host gut lumen, whereas Wolbachia and Rickettsia are usually scattered across insect tissues and cell types, including cells containing other symbionts, bacteriome sheath, fat body cells, gut epithelium, as well as hemolymph. However, there are exceptions, including Gammaproteobacteria that share bacteriome with Vidania, or Alphaproteobacteria that colonize Sulcia cells. We discuss how planthopper symbiont localization correlates with their acquisition and replacement patterns and the symbionts' likely functions. We also discuss the evolutionary consequences, constraints, and significance of these findings.}, } @article {pmid37032902, year = {2023}, author = {Wangwiwatsin, A and Kulwong, S and Phetcharaburanin, J and Namwat, N and Klanrit, P and Loilome, W and Maleewong, W and Reid, AJ}, title = {Toward novel treatment against filariasis: Insight into genome-wide co-evolutionary analysis of filarial nematodes and Wolbachia.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1052352}, pmid = {37032902}, issn = {1664-302X}, abstract = {Infectious diseases caused by filarial nematodes are major health problems for humans and animals globally. Current treatment using anti-helminthic drugs requires a long treatment period and is only effective against the microfilarial stage. Most species of filarial nematodes harbor a specific strain of Wolbachia bacteria, which are essential for the survival, development, and reproduction of the nematodes. This parasite-bacteria obligate symbiosis offers a new angle for the cure of filariasis. In this study, we utilized publicly available genome data and putative protein sequences from seven filarial nematode species and their symbiotic Wolbachia to screen for protein-protein interactions that could be a novel target against multiple filarial nematode species. Genome-wide in silico screening was performed to predict molecular interactions based on co-evolutionary signals. We identified over 8,000 pairs of gene families that show evidence of co-evolution based on high correlation score and low false discovery rate (FDR) between gene families and obtained a candidate list that may be keys in filarial nematode-Wolbachia interactions. Functional analysis was conducted on these top-scoring pairs, revealing biological processes related to various signaling processes, adult lifespan, developmental control, lipid and nucleotide metabolism, and RNA modification. Furthermore, network analysis of the top-scoring genes with multiple co-evolving pairs suggests candidate genes in both Wolbachia and the nematode that may play crucial roles at the center of multi-gene networks. A number of the top-scoring genes matched well to known drug targets, suggesting a promising drug-repurposing strategy that could be applicable against multiple filarial nematode species.}, } @article {pmid37027941, year = {2023}, author = {Liu, W and Nasir, M and Yan, M and Zhu, X and Iqbal, MS and Wang, L and Zhang, K and Li, D and Ji, J and Gao, X and Luo, J and Cui, J}, title = {Response of the Pardosa astrigera bacterial community to Cry1B protein.}, journal = {Ecotoxicology and environmental safety}, volume = {256}, number = {}, pages = {114855}, doi = {10.1016/j.ecoenv.2023.114855}, pmid = {37027941}, issn = {1090-2414}, mesh = {Female ; Male ; Humans ; Animals ; RNA, Ribosomal, 16S/metabolism ; *Bacteria ; *Spiders/metabolism ; Symbiosis ; Proteobacteria ; }, abstract = {While genetically modified (GM) crops bring economic benefits to human beings, their impact on non-target organisms has become an important part of environmental safety assessments. Symbiotic bacteria play an important role in eukaryotic biological functions and can adjust host communities to adapt to new environments. Therefore, this study examined the effects of Cry1B protein on the growth and development of non-target natural enemies of Pardosa astrigera (L. Koch) from the perspective of symbiotic bacteria. Cry1B protein had no significant effect on the health indicators of P. astrigera (adults and 2nd instar spiderlings). 16S rRNA sequencing results revealed that Cry1B protein did not change the symbiotic bacteria species composition of P. astrigera, but did reduce the number of OTU and species diversity. In 2nd instar spiderlings, neither the dominant phylum (Proteobacteria) nor the dominant genus (Acinetobacter) changed, but the relative abundance of Corynebacterium-1 decreased significantly; in adult spiders, the dominant bacteria genera of females and males were different. The dominant bacterial genera were Brevibacterium in females and Corynebacterium-1 in males, but Corynebacterium-1 was the dominant bacteria in both females and males feeding on Cry1B. The relative abundance of Wolbachia also increased significantly. In addition, bacteria in other genera varied significantly by sex. KEGG results showed that Cry1B protein only altered the significant enrichment of metabolic pathways in female spiders. In conclusion, the effects of Cry1B protein on symbiotic bacteria vary by growth and development stage and sex.}, } @article {pmid37022600, year = {2023}, author = {Caputo, B and Moretti, R and Virgillito, C and Manica, M and Lampazzi, E and Lombardi, G and Serini, P and Pichler, V and Beebe, NW and Della Torre, A and Calvitti, M}, title = {A bacterium against the tiger: further evidence of the potential of noninundative releases of males with manipulated Wolbachia infection in reducing fertility of Aedes albopictus field populations in Italy.}, journal = {Pest management science}, volume = {79}, number = {9}, pages = {3167-3176}, doi = {10.1002/ps.7495}, pmid = {37022600}, issn = {1526-4998}, support = {MUR PNRR Extended Partnership initiative on Emerging Infectious Diseases (project no. PE00000007, INF-ACT, research node 2).//Ministero della RIcerca/ ; }, mesh = {Animals ; Male ; Female ; *Aedes ; *Wolbachia ; Mosquito Control/methods ; Fertility ; Italy ; }, abstract = {BACKGROUND: Incompatible insect technique (IIT) is a population suppression approach based on the release of males with manipulated Wolbachia infection inducing egg inviability in wild females. We here present results of multiple field releases of incompatible ARwP males carried out in 2019 in a 2.7-ha green area within urban Rome (Italy) to assess the effect on Aedes albopictus egg viability. Data are compared with results obtained in 2018, when the approach was tested for the first time in Europe.

RESULTS: An average of 4674 ARwP males were released weekly for 7 weeks, resulting in a mean ARwP:wild male ratio of 1.1:1 (versus 0.7:1 in 2018). Egg-viability dynamics in ovitraps significantly varied between treated and control sites, with an estimated overall reduction of 35% (versus 15% in 2018). The estimated proportion of females classified as mated with ARwP males was 41.8% and the viability rate of eggs laid by these females (9.5%) was on average significantly lower than that of females only mated with wild males (87.8%); however, high variability in fertility was observed. Values of ARwP male competitiveness were 0.36 and 0.73 based on the overall viability rate of eggs in ovitraps and on female fertility, respectively; thus, well above the conventional 0.2 threshold for an effective suppressive impact in the field.

CONCLUSIONS: Results further support the potential of IIT as a tool to contribute to Ae. albopictus control in the urban context, stressing the need for larger field trials to evaluate the cost-efficacy of the approach in temperate regions. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.}, } @article {pmid37007535, year = {2023}, author = {Alomar, AA and Pérez-Ramos, DW and Kim, D and Kendziorski, NL and Eastmond, BH and Alto, BW and Caragata, EP}, title = {Native Wolbachia infection and larval competition stress shape fitness and West Nile virus infection in Culex quinquefasciatus mosquitoes.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1138476}, pmid = {37007535}, issn = {1664-302X}, abstract = {INTRODUCTION: Wolbachia transinfections established in key mosquito vectors, including Aedes aegypti are typically associated with pathogen blocking-reduced susceptibility to infection with key pathogens and reduced likelihood those pathogens are transmitted to new hosts. Host-symbiont-virus interactions are less well understood in mosquitoes like Culex quinquefasciatus, which naturally harbor Wolbachia, with pathogen blocking observed in some populations but not others, potentially due to innate differences in their Wolbachia load. In nature, mosquito larvae are often subject to developmental stresses associated with larval competition, which can lead to reduced body size and differential susceptibility to arbovirus infection.

METHODS: In this study, we sought to understand whether competition stress and Wolbachia infection in Cx. quinquefasciatus combine to impact host fitness and susceptibility to infection with West Nile virus. We reared Wolbachia-infected and uninfected Cx. quinquefasciatus larvae under three competition stress levels, increasing larval density without increasing the amount of food supplied. We then monitored larval development and survival, measured wing length and quantified Wolbachia density in adults, and then challenged mosquitoes from each treatment group orally with West Nile virus.

RESULTS AND DISCUSSION: We observed that high competition stress extended development time, decreased the likelihood of eclosion, decreased body size, and increased susceptibility to West Nile virus (WNV) infection. We also observed that Wolbachia infection reduced WNV load under low competition stress, and significantly improved the rate of survival for larval reared under higher competition stress. Consequently, our data suggest that native Wolbachia infection in Cx. quinquefasciatus has differential consequences for host fitness and susceptibility to WNV infection depending on competition stress.}, } @article {pmid37001324, year = {2023}, author = {Romano, DMM and Pereira, TN and Almeida, IB and Coelho, CSG and Duarte, FC and Harakava, R and Cassiano, LL and Mendes, MC}, title = {First molecular evidence of Wolbachia occurrence in Amblyomma sculptum (Acari: Ixodidae).}, journal = {Veterinary parasitology}, volume = {317}, number = {}, pages = {109907}, doi = {10.1016/j.vetpar.2023.109907}, pmid = {37001324}, issn = {1873-2550}, mesh = {Animals ; *Ixodidae/microbiology ; *Rocky Mountain Spotted Fever/epidemiology/microbiology/veterinary ; Amblyomma/genetics ; *Wolbachia/genetics ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Brazil/epidemiology ; *Ticks/microbiology ; *Rickettsia ; }, abstract = {As the main vector for the bacterium Rickettsia rickettsii in Brazil, the tick Amblyomma sculptum is a parasite of great public health importance in this country. Wolbachia is an endosymbiont bacterium highly widespread among invertebrates and because of its impact on its hosts' biology, form a powerful alternative for pests and disease control. The aim of this study was to investigate the occurrence of this bacterium in A. sculptum. For this, 187 adult ticks collected in two municipalities in the interior of the state of São Paulo, Brazil, were analyzed using molecular techniques and bioinformatics tools. A total of 15 ticks were positive for the presence of Wolbachia. Phylogenetic analysis on the 16S rRNA gene indicated that the Wolbachia DNA sequences obtained in this investigation belonged to different clades, probably in supergroups B and F. This was the first study to report the occurrence of Wolbachia in A. sculptum and it enriches knowledge about the susceptibility of ticks to this bacterium. Now that we know that Wolbachia can be found in A. sculptum, the objective for a next study must be to investigate Wolbachia's possible origin in this tick.}, } @article {pmid37000335, year = {2023}, author = {Rohlfing, K and Yue, L and Franke, S and Zeng, C and Podsiadlowski, L and Dobler, S}, title = {When does the female bias arise? Insights from the sex determination cascade of a flea beetle with a strongly skewed sex ratio.}, journal = {Functional & integrative genomics}, volume = {23}, number = {2}, pages = {112}, pmid = {37000335}, issn = {1438-7948}, mesh = {Animals ; Male ; Female ; *Coleoptera/genetics ; Sex Ratio ; *Siphonaptera/genetics ; Larva ; DNA, Mitochondrial ; RNA, Messenger ; }, abstract = {Reproduction-manipulating bacteria like Wolbachia can shift sex ratios in insects towards females, but skewed sex ratios may also arise from genetic conflicts. The flea beetle Altica lythri harbors three main mtDNA strains that are coupled to three different Wolbachia infections. Depending on the mtDNA types, the females produce either offspring with a balanced sex ratio or exclusively daughters. To obtain markers that can monitor when sex bias arises in the beetle's ontogeny, we elucidated the sex determination cascade of A. lythri. We established a RT-PCR method based on length variants of dsx (doublesex) transcripts to determine the sex of morphologically indistinguishable eggs and larvae. In females of one mtDNA type (HT1/HT1*) known to produce only daughters, male offspring were already missing at the egg stage while for females of another type (HT2), the dsx splice variants revealed a balanced sex ratio among eggs and larvae. Our data suggest that the sex determination cascade in A. lythri is initiated by maternally transmitted female-specific tra (transformer) mRNA as primary signal. This tra mRNA seems to be involved in a positive feedback loop that maintains the production of the female splice variant, as known for female offspring in Tribolium castaneum. The translation of the maternally transmitted female tra mRNA must be inhibited in male offspring, but the underlying primary genetic signal remains to be identified. We discuss which differences between the mtDNA types can influence sex determination and lead to the skewed sex ratio of HT1.}, } @article {pmid36996252, year = {2023}, author = {Hornett, EA and Hurst, GDD}, title = {One strain may hide another: Cryptic male-killing Wolbachia.}, journal = {PLoS biology}, volume = {21}, number = {3}, pages = {e3002076}, pmid = {36996252}, issn = {1545-7885}, mesh = {Animals ; Male ; *Wolbachia ; Insecta ; Host-Pathogen Interactions ; Host Microbial Interactions ; }, abstract = {While heritable symbionts are common in insects, strains that act as male-killers are considered rare. A new study in PLOS Biology identifies a novel male-killer hidden by coinfection and host resistance, highlighting the complexity of host-microbial interactions in natural systems.}, } @article {pmid36993585, year = {2023}, author = {Beckmann, J and Gillespie, J and Tauritz, D}, title = {Modelling Emergence of Wolbachia Toxin-Antidote Protein Functions with an Evolutionary Algorithm.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36993585}, support = {R21 AI146773/AI/NIAID NIH HHS/United States ; R21 AI156762/AI/NIAID NIH HHS/United States ; R21 AI166832/AI/NIAID NIH HHS/United States ; }, abstract = {Evolutionary algorithms (EAs) simulate Darwinian evolution and adeptly mimic natural evolution. Most EA applications in biology encode high levels of abstraction in top-down ecological population models. In contrast, our research merges protein alignment algorithms from bioinformatics into codon based EAs that simulate molecular protein string evolution from the bottom up. We apply our EA to reconcile a problem in the field of Wolbachia induced cytoplasmic incompatibility (CI). Wolbachia is a microbial endosymbiont that lives inside insect cells. CI is conditional insect sterility that operates as a toxin antidote (TA) system. Although, CI exhibits complex phenotypes not fully explained under a single discrete model. We instantiate in-silico genes that control CI, CI factors (cifs), as strings within the EA chromosome. We monitor the evolution of their enzymatic activity, binding, and cellular localization by applying selective pressure on their primary amino acid strings. Our model helps rationalize why two distinct mechanisms of CI induction might coexist in nature. We find that nuclear localization signals (NLS) and Type IV secretion system signals (T4SS) are of low complexity and evolve fast, whereas binding interactions have intermediate complexity, and enzymatic activity is the most complex. Our model predicts that as ancestral TA systems evolve into eukaryotic CI systems, the placement of NLS or T4SS signals can stochastically vary, imparting effects that might impact CI induction mechanics. Our model highlights how preconditions, genetic diversity, and sequence length can bias evolution of cifs towards one mechanism or another.}, } @article {pmid36992487, year = {2023}, author = {Ferreira, QR and Lemos, FFB and Moura, MN and Nascimento, JOS and Novaes, AF and Barcelos, IS and Fernandes, LA and Amaral, LSB and Barreto, FK and Melo, FF}, title = {Role of the Microbiome in Aedes spp. Vector Competence: What Do We Know?.}, journal = {Viruses}, volume = {15}, number = {3}, pages = {}, pmid = {36992487}, issn = {1999-4915}, mesh = {Animals ; Female ; Humans ; *Aedes ; *Chikungunya Fever ; *Arboviruses/genetics ; *Yellow Fever ; *Zika Virus Infection ; *Zika Virus ; *Microbiota ; }, abstract = {Aedes aegypti and Aedes albopictus are the vectors of important arboviruses: dengue fever, chikungunya, Zika, and yellow fever. Female mosquitoes acquire arboviruses by feeding on the infected host blood, thus being able to transmit it to their offspring. The intrinsic ability of a vector to infect itself and transmit a pathogen is known as vector competence. Several factors influence the susceptibility of these females to be infected by these arboviruses, such as the activation of the innate immune system through the Toll, immunodeficiency (Imd), JAK-STAT pathways, and the interference of specific antiviral response pathways of RNAi. It is also believed that the presence of non-pathogenic microorganisms in the microbiota of these arthropods could influence this immune response, as it provides a baseline activation of the innate immune system, which may generate resistance against arboviruses. In addition, this microbiome has direct action against arboviruses, mainly due to the ability of Wolbachia spp. to block viral genome replication, added to the competition for resources within the mosquito organism. Despite major advances in the area, studies are still needed to evaluate the microbiota profiles of Aedes spp. and their vector competence, as well as further exploration of the individual roles of microbiome components in activating the innate immune system.}, } @article {pmid36989319, year = {2023}, author = {Wimalasiri-Yapa, BMCR and Huang, B and Ross, PA and Hoffmann, AA and Ritchie, SA and Frentiu, FD and Warrilow, D and van den Hurk, AF}, title = {Differences in gene expression in field populations of Wolbachia-infected Aedes aegypti mosquitoes with varying release histories in northern Australia.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {3}, pages = {e0011222}, pmid = {36989319}, issn = {1935-2735}, mesh = {Animals ; *Aedes ; *Dengue Virus/physiology ; *Wolbachia/genetics ; Mosquito Vectors ; *Zika Virus/genetics ; *Zika Virus Infection ; Australia ; Gene Expression ; }, abstract = {Aedes aegypti is the principal mosquito vector of dengue, yellow fever, Zika and chikungunya viruses. The wMel strain of the endosymbiotic bacteria Wolbachia pipientis was introduced into the vector as a novel biocontrol strategy to stop transmission of these viruses. Mosquitoes with Wolbachia have been released in the field in Northern Queensland, Australia since 2011, at various locations and over several years, with populations remaining stably infected. Wolbachia infection is known to alter gene expression in its mosquito host, but whether (and how) this changes over the long-term in the context of field releases remains unknown. We sampled mosquitoes from Wolbachia-infected populations with three different release histories along a time gradient and performed RNA-seq to investigate gene expression changes in the insect host. We observed a significant impact on gene expression in Wolbachia-infected mosquitoes versus uninfected controls. Fewer genes had significantly upregulated expression in mosquitoes from the older releases (512 and 486 from the 2011 and 2013/14 release years, respectively) versus the more recent releases (1154 from the 2017 release year). Nonetheless, a fundamental signature of Wolbachia infection on host gene expression was observed across all releases, comprising upregulation of immunity (e.g. leucine-rich repeats, CLIPs) and metabolism (e.g. lipid metabolism, iron transport) genes. There was limited downregulation of gene expression in mosquitoes from the older releases (84 and 71 genes from the 2011 and 2013/14 release years, respectively), but significantly more in the most recent release (509 from the 2017 release year). Our findings indicate that at > 8 years post-introgression into field populations, Wolbachia continues to profoundly impact expression of host genes, such as those involved in insect immune response and metabolism. If Wolbachia-mediated virus blocking is underpinned by these differential gene expression changes, our results suggest it may remain stable long-term.}, } @article {pmid36986288, year = {2023}, author = {Moore, C and Lashnits, E and Neupane, P and Herrin, BH and Lappin, M and André, MR and Breitschwerdt, EB}, title = {Feeding on a Bartonella henselae Infected Host Triggers Temporary Changes in the Ctenocephalides felis Microbiome.}, journal = {Pathogens (Basel, Switzerland)}, volume = {12}, number = {3}, pages = {}, pmid = {36986288}, issn = {2076-0817}, support = {FAPESP Process 2019/09464-6//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; }, abstract = {The effect of Bartonella henselae on the microbiome of its vector, Ctenocephalides felis (the cat flea) is largely unknown, as the majority of C. felis microbiome studies have utilized wild-caught pooled fleas. We surveyed the microbiome of laboratory-origin C. felis fed on B. henselae-infected cats for 24 h or 9 days to identify changes to microbiome diversity and microbe prevalence compared to unfed fleas, and fleas fed on uninfected cats. Utilizing Next Generation Sequencing (NGS) on the Illumina platform, we documented an increase in microbial diversity in C. felis fed on Bartonella-infected cats for 24 h. These changes returned to baseline (unfed fleas or fleas fed on uninfected cats) after 9 days on the host. Increased diversity in the C. felis microbiome when fed on B. henselae-infected cats may be related to the mammalian, flea, or endosymbiont response. Poor B. henselae acquisition was documented with only one of four infected flea pools having B. henselae detected by NGS. We hypothesize this is due to the use of adult fleas, flea genetic variation, or lack of co-feeding with B. henselae-infected fleas. Future studies are necessary to fully characterize the effect of endosymbionts and C. felis diversity on B. henselae acquisition.}, } @article {pmid36985370, year = {2023}, author = {Remmal, I and Bel Mokhtar, N and Maurady, A and Reda Britel, M and El Fakhouri, K and Asimakis, E and Tsiamis, G and Stathopoulou, P}, title = {Characterization of the Bacterial Microbiome in Natural Populations of Barley Stem Gall Midge, Mayetiola hordei, in Morocco.}, journal = {Microorganisms}, volume = {11}, number = {3}, pages = {}, pmid = {36985370}, issn = {2076-2607}, support = {MIS 5010952//Hellenic Ministry of Rural Development and Food/ ; 00//European Union Erasmus+ Programme/ ; }, abstract = {Mayetiola hordei (Kieffer), known as barley stem gall midge, is one of the most destructive barley pests in many areas around the world, inflicting significant qualitative and quantitative damage to crop production. In this study, we investigate the presence of reproductive symbionts, the effect of geographical origin on the bacterial microbiome's structure, and the diversity associated with natural populations of M. hordei located in four barley-producing areas in Morocco. Wolbachia infection was discovered in 9% of the natural populations using a precise 16S rDNA PCR assay. High-throughput sequencing of the V3-V4 region of the bacterial 16S rRNA gene indicated that the native environments of samples had a substantial environmental impact on the microbiota taxonomic assortment. Briefly, 5 phyla, 7 classes, and 42 genera were identified across all the samples. To our knowledge, this is the first report on the bacterial composition of M. hordei natural populations. The presence of Wolbachia infection may assist in the diagnosis of ideal natural populations, providing a new insight into the employment of Wolbachia in the control of barley midge populations, in the context of the sterile insect technique or other biological control methods.}, } @article {pmid36985289, year = {2023}, author = {Huynh, LN and Diarra, AZ and Pham, QL and Berenger, JM and Ho, VH and Nguyen, XQ and Parola, P}, title = {Identification of Vietnamese Flea Species and Their Associated Microorganisms Using Morphological, Molecular, and Protein Profiling.}, journal = {Microorganisms}, volume = {11}, number = {3}, pages = {}, pmid = {36985289}, issn = {2076-2607}, abstract = {Fleas are obligatory blood-sucking ectoparasites of medical and veterinary importance. The identification of fleas and associated flea-borne microorganisms, therefore, plays an important role in controlling and managing these vectors. Recently, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been reported as an innovative and effective approach to the identification of arthropods, including fleas. This study aims to use this technology to identify ethanol-preserved fleas collected in Vietnam and to use molecular biology to search for microorganisms associated with these fleas. A total of 502 fleas were collected from wild and domestic animals in four provinces in Vietnam. Morphological identification led to the recognition of five flea species, namely Xenopsylla cheopis, Xenopsylla astia, Pulex irritans, Ctenocephalides canis, and Ctenocephalides felis. The cephalothoraxes of 300 individual, randomly selected fleas were tested using MALDI-TOF MS and molecular analysis for the identification and detection of microorganisms. A total of 257/300 (85.7%) of the obtained spectra from the cephalothoraxes of each species were of good enough quality to be used for our analyses. Our laboratory MALDI-TOF MS reference database was upgraded with spectra achieved from five randomly selected fleas for every species of Ctenocephalides canis and Ctenocephalides felis. The remaining spectra were then queried against the upgraded MALDI-TOF MS database, which showed 100% correspondence between morphology and MALDI-TOF MS identification for two flea species (Ctenocephalides canis and Ctenocephalides felis). The MS spectra of the remaining species (three P. irritans, five X. astia, and two X. cheopis) were visually generated low-intensity MS profiles with high background noise that could not be used to update our database. Bartonella and Wolbachia spp. were detected in 300 fleas from Vietnam using PCR and sequencing with primers derived from the gltA gene for Bartonella and the 16S rRNA gene for Wolbachia, including 3 Bartonella clarridgeiae (1%), 3 Bartonella rochalimae (1%), 1 Bartonella coopersplainsensis (0.3%), and 174 Wolbachia spp. endosymbionts (58%).}, } @article {pmid36977163, year = {2023}, author = {Florez, D and Young, AJ and Bernabé, KJ and Hyman, JM and Qu, Z}, title = {Modeling Sustained Transmission of Wolbachia among Anopheles Mosquitoes: Implications for Malaria Control in Haiti.}, journal = {Tropical medicine and infectious disease}, volume = {8}, number = {3}, pages = {}, pmid = {36977163}, issn = {2414-6366}, support = {New Faculty Startup Funds//The University of Texas at San Antonio/ ; }, abstract = {Wolbachia infection in Anopheles albimanus mosquitoes can render mosquitoes less capable of spreading malaria. We developed and analyzed a mechanistic compartmental ordinary differential equation model to evaluate the effectiveness of Wolbachia-based vector control strategies among wild Anopheles mosquitoes in Haiti. The model tracks the mosquito life stages, including egg, larva, and adult (male and female). It also accounts for critical biological effects, such as the maternal transmission of Wolbachia through infected females and cytoplasmic incompatibility, which effectively sterilizes uninfected females when they mate with infected males. We derive and interpret dimensionless numbers, including the basic reproductive number and next-generation numbers. The proposed system presents a backward bifurcation, which indicates a threshold infection that needs to be exceeded to establish a stable Wolbachia infection. The sensitivity analysis ranks the relative importance of the epidemiological parameters at baseline. We simulate different intervention scenarios, including prerelease mitigation using larviciding and thermal fogging before the release, multiple releases of infected populations, and different release times of the year. Our simulations show that the most efficient approach to establishing Wolbachia is to release all the infected mosquitoes immediately after the prerelease mitigation process. Moreover, the model predicts that it is more efficient to release during the dry season than the wet season.}, } @article {pmid36975996, year = {2023}, author = {Jiang, RX and Shang, F and Jiang, HB and Dou, W and Cernava, T and Wang, JJ}, title = {Environmental Factors Affect the Bacterial Community in Diaphorina citri, an Important Vector of "Candidatus Liberibacter asiaticus".}, journal = {Microbiology spectrum}, volume = {11}, number = {2}, pages = {e0529822}, pmid = {36975996}, issn = {2165-0497}, abstract = {Insects are associated with diverse microbial communities that can have substantial effects on hosts. Here, we characterized the bacterial communities in the Asian citrus psyllid (ACP), Diaphorina citri (Hemiptera: Psyllidae), a major vector of the devastating pathogen "Candidatus Liberibacter asiaticus," which causes citrus Huanglongbing (HLB). In total, 256 ACP individuals across 15 field sites and one laboratory population in China were sequenced. The results showed that the bacterial community diversity was the highest in the Guilin population (average Shannon index, 1.27), and the highest value for richness was found in the Chenzhou population (average Chao1 index, 298). The bacterial community structures of the field-collected populations were significantly different, and all of them harbored Wolbachia, which was assigned to strain ST-173. Structural equation models revealed that the dominant Wolbachia strain had a significantly negative correlation with the annual mean temperature. In addition, the results obtained with populations infected with "Ca. Liberibacter asiaticus" indicated that in total, 140 bacteria could be involved in interactions with this bacterium. The ACP field populations harbored a more diverse bacterial community than the laboratory population, and the relative occurrences of some symbionts differed significantly. However, the bacterial community of the ACP laboratory colony was connected in a more complex network structure (average degree, 54.83) than that of the field populations (average degree, 10.62). Our results provide evidence that environmental factors can influence the bacterial community structure and bacterial relative abundance in ACP populations. This is likely due to the adaptation of ACPs to local environments. IMPORTANCE The Asian citrus psyllid (ACP) is an important vector of the HLB pathogen, which is a major threat to citrus production around the world. Bacterial communities harbored by insects could be affected by different environmental factors. Understanding these factors that affect the bacterial community of the ACP could be important for the better management of HLB transmission. This work surveyed ACP field populations in mainland China in order to explore the bacterial community diversity of different populations and the potential relationships between environmental factors and predominant symbionts. We have assessed the differences in ACP bacterial communities and identified the prevalent Wolbachia strains in the field. In addition, we compared the bacterial communities of ACP field-collected and laboratory populations. Comparing populations subjected to contrasting conditions could help us to better understand how the ACP adapts to local environmental conditions. Our study provides new insights into how environmental factors influence the bacterial community of the ACP.}, } @article {pmid36958642, year = {2023}, author = {Detcharoen, M and Jiggins, FM and Schlick-Steiner, BC and Steiner, FM}, title = {Wolbachia endosymbiotic bacteria alter the gut microbiome in the fly Drosophila nigrosparsa.}, journal = {Journal of invertebrate pathology}, volume = {198}, number = {}, pages = {107915}, doi = {10.1016/j.jip.2023.107915}, pmid = {36958642}, issn = {1096-0805}, mesh = {Animals ; Drosophila/microbiology ; *Wolbachia ; *Gastrointestinal Microbiome ; Reproduction ; Bacteria ; Symbiosis ; Drosophila melanogaster/microbiology ; }, abstract = {Wolbachia are known to cause reproductive manipulations and in some arthropod species, Wolbachia were reported to cause changes in gut microbiome. However, the effects of Wolbachia bacteria on the microbiomes of their hosts, including Drosophila flies, have not been fully accessed. Here, we checked the bacterial microbiome in guts of Wolbachia-uninfected and of Wolbachia-infected Drosophila nigrosparsa, both separated into a bleach-only (embryos bleached) and a gnotobiotic (embryos bleached and inoculated with bacteria) treatment. We observed a clear separation between the Wolbachia-infected and the Wolbachia-uninfected samples, and the infected samples had higher variation in alpha diversity than the uninfected ones. There were reductions in the abundances of Proteobacteria (Pseudomonadota), especially Acetobacter, in the infected samples of both treatments. These findings highlight that Wolbachia change the gut microbiome in D. nigrosparsa as well as that the interactions between Wolbachia and bacteria like Acetobacter need to be investigated.}, } @article {pmid36958587, year = {2023}, author = {Verhulst, EC and Pannebakker, BA and Geuverink, E}, title = {Variation in sex determination mechanisms may constrain parthenogenesis-induction by endosymbionts in haplodiploid systems.}, journal = {Current opinion in insect science}, volume = {56}, number = {}, pages = {101023}, doi = {10.1016/j.cois.2023.101023}, pmid = {36958587}, issn = {2214-5753}, mesh = {Animals ; Female ; Parthenogenesis ; *Wolbachia ; *Arthropods ; Sex Determination Processes ; }, abstract = {Endosymbionts are maternally transmitted, and therefore benefit from maximizing female offspring numbers. Parthenogenesis-induction (PI) is the most effective type of manipulation for transmission, but has solely been detected in haplodiploid species, whereas cytoplasmic incompatibility (CI) is detected frequently across the arthropod phylum, including haplodiploids. This puzzling observation led us to hypothesize that the molecular sex-determination mechanism of the haplodiploid host may be a constraining factor in the ability of endosymbionts to induce parthenogenesis. Recent insights indicate that PI-endosymbionts may be able to directly manipulate sex-determination genes to induce the necessary steps required for PI in haplodiploids. However, sex-determination cascades vary extensively, so PI-induction would require a specialized and host-dependent tool set. Contrastingly, CI-related genes target conserved cell-cycle mechanisms, are located on mobile elements, and spread easily. Finally, endosymbiont-manipulations may have a strong impact on the effectiveness of haplodiploid biocontrol agents, but can also be used to enhance their efficacy.}, } @article {pmid36950155, year = {2023}, author = {Moore, C and Breitschwerdt, EB and Kim, L and Li, Y and Ferris, K and Maggi, R and Lashnits, E}, title = {The association of host and vector characteristics with Ctenocephalides felis pathogen and endosymbiont infection.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1137059}, pmid = {36950155}, issn = {1664-302X}, abstract = {Surveillance of the fleas and flea-borne pathogens infecting cats is important for both human and animal health. Multiple zoonotic Bartonella and Rickettsia species are known to infect the most common flea infesting cats and dogs worldwide: Ctenocephalides felis, the cat flea. The ability of other flea species to transmit pathogens is relatively unexplored. We aimed to determine cat host and flea factors independently associated with flea Bartonella and Rickettsia infection. We also assessed flea and cat infection by flea-host pair and location. To accomplish these aims, we performed qPCR for the detection of Bartonella, hemotropic Mycoplasma, Rickettsia, and Wolbachia DNA using paired cat and flea samples obtained from free-roaming cats presenting for spay or neuter across four locations in the United States. A logistic regression model was employed to identify the effect of cat (sex, body weight, geographic location, and Bartonella, hemotropic Mycoplasma, and Rickettsia spp., infection) and flea (clade and Rickettsia and Wolbachia infection) factors on C. felis Bartonella clarridgeiae infection. From 189 free roaming cats, we collected 84 fleas: Ctenocephalides felis (78/84), Cediopsylla simplex (4/84), Orchopeas howardi (1/84), and Nosopsyllus fasciatus (1/84). Ctenocephalides felis were phylogenetically assigned to Clades 1, 4, and 6 by cox1 gene amplification. Rickettsia asembonensis (52/84) and B. clarridgeiae (16/84) were the most common pathogenic bacteria detected in fleas. Our model identified host cat sex and weight as independently associated with B. clarridgeiae infection in fleas. Rickettsia asembonensis (52/84), Rickettsia felis (7/84) and Bartonella henselae (7/84) were detected in specific clades: R. felis was detected only in Clades 1 and 6 while B. henselae and R. asembonensis were detected only in Clade 4. Wolbachia spp., also displayed clade specificity with strains other than Wolbachia wCfeT only infecting fleas from Clade 6. There was poor flea and host agreement for Bartonella spp., infection; however, there was agreement in the Bartonella species detected in cats and fleas by geographic location. These findings reinforce the importance of considering reservoir host attributes and vector phylogenetic diversity in epidemiological studies of flea-borne pathogens. Widespread sampling is necessary to identify the factors driving flea-borne pathogen presence and transmission.}, } @article {pmid36949814, year = {2023}, author = {Ou, D and Qiu, JH and Su, ZQ and Wang, L and Qiu, BL}, title = {The phylogeny and distribution of Wolbachia in two pathogen vector insects, Asian citrus psyllid and Longan psyllid.}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1121186}, pmid = {36949814}, issn = {2235-2988}, mesh = {Animals ; *Hemiptera/microbiology ; *Wolbachia/genetics ; Phylogeny ; *Citrus/microbiology ; }, abstract = {BACKGROUND: Wolbachia is the most abundant bacterial endosymbiont among insects. It can play a prominent role in the development, reproduction and immunity of its given insect host. To date, Wolbachia presence is well studied within aphids, whiteflies and planthoppers, but relatively few studies have investigated its presence in psyllids.

METHODS: Here, the infection status of Wolbachia in five species of psyllid, including Asian citrus psyllid Diaphorina citri and longan psyllid Cornegenapsylla sinica was investigated. The phylogenetic relationships of different Wolbachia lines and their infection density and patterns in D. citri and C. sinica from different countries was also examined.

RESULTS: The infection rates of Wolbachia in D. citri and C. sinica were both 100%, and their sequencing types are ST173 and ST532 respectively. Phylogenetic analysis revealed that the Wolbachia lines in D. citri and C. sinica both belong to the Con subgroup of Wolbachia supergroup B. In addition, Wolbachia displayed a scattered localization pattern in the 5th instar nymphs and in the reproductive organs of both D. citri and C. sinica but differed in other tissues; it was highest in the midgut, lowest in the salivary glands and medium in both the testes and ovaries.

CONCLUSION: Our findings assist in further understanding the coevolution of Wolbachia and its psyllid hosts. Given that Wolbachia could play an important role in insect pest control and pathogen transmission inhibition, our findings may also provide new insights for development of control strategies for D. citri and C. sinica.}, } @article {pmid36947551, year = {2023}, author = {Bourne, ME and Gloder, G and Weldegergis, BT and Slingerland, M and Ceribelli, A and Crauwels, S and Lievens, B and Jacquemyn, H and Dicke, M and Poelman, EH}, title = {Parasitism causes changes in caterpillar odours and associated bacterial communities with consequences for host-location by a hyperparasitoid.}, journal = {PLoS pathogens}, volume = {19}, number = {3}, pages = {e1011262}, pmid = {36947551}, issn = {1553-7374}, mesh = {Animals ; Odorants ; Larva ; *Butterflies/parasitology ; *Wasps/parasitology ; Host-Parasite Interactions ; }, abstract = {Microorganisms living in and on macroorganisms may produce microbial volatile compounds (mVOCs) that characterise organismal odours. The mVOCs might thereby provide a reliable cue to carnivorous enemies in locating their host or prey. Parasitism by parasitoid wasps might alter the microbiome of their caterpillar host, affecting organismal odours and interactions with insects of higher trophic levels such as hyperparasitoids. Hyperparasitoids parasitise larvae or pupae of parasitoids, which are often concealed or inconspicuous. Odours of parasitised caterpillars aid them to locate their host, but the origin of these odours and its relationship to the caterpillar microbiome are unknown. Here, we analysed the odours and microbiome of the large cabbage white caterpillar Pieris brassicae in relation to parasitism by its endoparasitoid Cotesia glomerata. We identified how bacterial presence in and on the caterpillars is correlated with caterpillar odours and tested the attractiveness of parasitised and unparasitised caterpillars to the hyperparasitoid Baryscapus galactopus. We manipulated the presence of the external microbiome and the transient internal microbiome of caterpillars to identify the microbial origin of odours. We found that parasitism by C. glomerata led to the production of five characteristic volatile products and significantly affected the internal and external microbiome of the caterpillar, which were both found to have a significant correlation with caterpillar odours. The preference of the hyperparasitoid was correlated with the presence of the external microbiome. Likely, the changes in external microbiome and body odour after parasitism were driven by the resident internal microbiome of caterpillars, where the bacterium Wolbachia sp. was only present after parasitism. Micro-injection of Wolbachia in unparasitised caterpillars increased hyperparasitoid attraction to the caterpillars compared to untreated caterpillars, while no differences were found compared to parasitised caterpillars. In conclusion, our results indicate that host-parasite interactions can affect multi-trophic interactions and hyperparasitoid olfaction through alterations of the microbiome.}, } @article {pmid36947547, year = {2023}, author = {Richardson, KM and Ross, PA and Cooper, BS and Conner, WR and Schmidt, TL and Hoffmann, AA}, title = {A male-killing Wolbachia endosymbiont is concealed by another endosymbiont and a nuclear suppressor.}, journal = {PLoS biology}, volume = {21}, number = {3}, pages = {e3001879}, pmid = {36947547}, issn = {1545-7885}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Male ; *Wolbachia/genetics ; Reproduction ; Drosophila/genetics ; Phenotype ; Insecta ; Symbiosis ; }, abstract = {Bacteria that live inside the cells of insect hosts (endosymbionts) can alter the reproduction of their hosts, including the killing of male offspring (male killing, MK). MK has only been described in a few insects, but this may reflect challenges in detecting MK rather than its rarity. Here, we identify MK Wolbachia at a low frequency (around 4%) in natural populations of Drosophila pseudotakahashii. MK Wolbachia had a stable density and maternal transmission during laboratory culture, but the MK phenotype which manifested mainly at the larval stage was lost rapidly. MK Wolbachia occurred alongside a second Wolbachia strain expressing a different reproductive manipulation, cytoplasmic incompatibility (CI). A genomic analysis highlighted Wolbachia regions diverged between the 2 strains involving 17 genes, and homologs of the wmk and cif genes implicated in MK and CI were identified in the Wolbachia assembly. Doubly infected males induced CI with uninfected females but not females singly infected with CI-causing Wolbachia. A rapidly spreading dominant nuclear suppressor genetic element affecting MK was identified through backcrossing and subsequent analysis with ddRAD SNPs of the D. pseudotakahashii genome. These findings highlight the complexity of nuclear and microbial components affecting MK endosymbiont detection and dynamics in populations and the challenges of making connections between endosymbionts and the host phenotypes affected by them.}, } @article {pmid36944122, year = {2023}, author = {Čisovská Bazsalovicsová, E and Víchová, B and Oboňa, J and Radačovská, A and Blažeková, V and Králová-Hromadová, I}, title = {Bird Louse Flies Ornithomya spp. (Diptera: Hippoboscidae) as Potential Vectors of Mammalian Babesia and Other Pathogens.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {23}, number = {5}, pages = {275-283}, doi = {10.1089/vbz.2022.0088}, pmid = {36944122}, issn = {1557-7759}, mesh = {Humans ; Male ; Female ; Animals ; *Babesia/genetics ; *Diptera ; *Rickettsia/genetics ; Birds ; Mammals ; *Anoplura ; }, abstract = {Background: Birds and mammals share various ectoparasites, which are responsible for the transmission of a wide range of pathogens. The louse flies (family Hippoboscidae) are ectoparasitic dipterans feeding strictly on the blood of mammals and birds. Both sexes of the louse flies are obligatory hematophagous and are known to act as the vectors of infectious agents. Materials and Methods: A total of 20 specimens of Ornithomya sp. were collected by hand on birds caught in nets or by hand from humans in two localities in Eastern Slovakia in 2021. The DNA samples were individually screened by species-specific PCRs for the presence of selected vector-borne pathogens. Results: Taxonomic identification folowed by molecular analyses revealed two louse fly species of Ornithomya spp. (O. avicularia and O. biloba). The molecular screening provided negative PCR results for Anaplasma phagocytophilum, Borrelia burgdorferi s.l., Rickettsia spp., Bartonella spp., and Hepatozoon canis. In contrast, positive PCR results were obtained for Babesia spp., Wolbachia spp., and Trypanosoma corvi. Conclusions: Of epidemiological importance is that the louse flies can presumably spread Babesia and other pathogens by host switching which facilitates the transmission and spread of numerous pathogens.}, } @article {pmid36934294, year = {2023}, author = {Allman, MJ and Lin, YH and Joubert, DA and Addley-Cook, J and Mejía-Torres, MC and Simmons, CP and Flores, HA and Fraser, JE}, title = {Enhancing the scalability of Wolbachia-based vector-borne disease management: time and temperature limits for storage and transport of Wolbachia-infected Aedes aegypti eggs for field releases.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {108}, pmid = {36934294}, issn = {1756-3305}, mesh = {Animals ; Temperature ; *Aedes ; Mosquito Vectors ; *Wolbachia ; Eggs ; }, abstract = {BACKGROUND: Introgression of the bacterial endosymbiont Wolbachia into Aedes aegypti populations is a biocontrol approach being used to reduce arbovirus transmission. This requires mass release of Wolbachia-infected mosquitoes. While releases have been conducted using a variety of techniques, egg releases, using water-soluble capsules containing mosquito eggs and larval food, offer an attractive method due to its potential to reduce onsite resource requirements. However, optimisation of this approach is required to ensure there is no detrimental impact on mosquito fitness and to promote successful Wolbachia introgression.

METHODS: We determined the impact of storage time and temperature on wild-type (WT) and Wolbachia-infected (wMel or wAlbB strains) Ae. aegypti eggs. Eggs were stored inside capsules over 8 weeks at 18 °C or 22 °C and hatch rate, emergence rate and Wolbachia density were determined. We next examined egg quality and Wolbachia density after exposing eggs to 4-40 °C to determine how eggs may be impacted if exposed to extreme temperatures during shipment.

RESULTS: Encapsulating eggs for 8 weeks did not negatively impact egg viability or resulting adult emergence and Wolbachia density compared to controls. When eggs were exposed to temperatures within 4-36 °C for 48 h, their viability and resulting adult Wolbachia density were maintained; however, both were significantly reduced when exposed to 40 °C.

CONCLUSIONS: We describe the time and temperature limits for maintaining viability of Wolbachia-infected Ae. aegypti eggs when encapsulated or exposed to extreme temperatures. These findings could improve the efficiency of mass releases by providing transport and storage constraints to ensure only high-quality material is utilised during field releases.}, } @article {pmid36933571, year = {2023}, author = {Oladipupo, SO and Carroll, JD and Beckmann, JF}, title = {Convergent Aedes and Drosophila CidB interactomes suggest cytoplasmic incompatibility targets are conserved.}, journal = {Insect biochemistry and molecular biology}, volume = {155}, number = {}, pages = {103931}, doi = {10.1016/j.ibmb.2023.103931}, pmid = {36933571}, issn = {1879-0240}, mesh = {Animals ; Male ; Drosophila ; Drosophila melanogaster ; *Aedes ; Semen ; Cytoplasm/metabolism ; *Wolbachia ; }, abstract = {Wolbachia-mediated cytoplasmic incompatibility (CI) is a conditional embryonic lethality induced when Wolbachia-modified sperm fertilizes an uninfected egg. The Wolbachia proteins, CidA and CidB control CI. CidA is a rescue factor that reverses lethality. CidA binds to CidB. CidB contains a deubiquitinating enzyme and induces CI. Precisely how CidB induces CI and what it targets are unknown. Likewise, how CidA prevents sterilization by CidB is not clear. To identify CidB substrates in mosquitos we conducted pull-down assays using recombinant CidA and CidB mixed with Aedes aegypti lysates to identify the protein interactomes of CidB and the CidB/CidA protein complex. Our data allow us to cross compare CidB interactomes across taxa for Aedes and Drosophila. Our data replicate several convergent interactions, suggesting that CI targets conserved substrates across insects. Our data support a hypothesis that CidA rescues CI by tethering CidB away from its substrates. Specifically, we identify ten convergent candidate substrates including P32 (protamine-histone exchange factor), karyopherin alpha, ubiquitin-conjugating enzyme, and bicoid stabilizing factor. Future analysis on how these candidates contribute to CI will clarify mechanisms.}, } @article {pmid36929176, year = {2023}, author = {Eugénio, AT and Marialva, MSP and Beldade, P}, title = {Effects of Wolbachia on Transposable Element Expression Vary Between Drosophila melanogaster Host Genotypes.}, journal = {Genome biology and evolution}, volume = {15}, number = {3}, pages = {}, pmid = {36929176}, issn = {1759-6653}, mesh = {Animals ; *Drosophila melanogaster/genetics ; DNA Transposable Elements ; *Wolbachia/genetics ; Evolution, Molecular ; Genotype ; }, abstract = {Transposable elements (TEs) are repetitive DNA sequences capable of changing position in host genomes, thereby causing mutations. TE insertions typically have deleterious effects but they can also be beneficial. Increasing evidence of the contribution of TEs to adaptive evolution further raises interest in understanding what factors impact TE activity. Based on previous studies associating the bacterial endosymbiont Wolbachia with changes in the abundance of piRNAs, a mechanism for TE repression, and to transposition of specific TEs, we hypothesized that Wolbachia infection would interfere with TE activity. We tested this hypothesis by studying the expression of 14 TEs in a panel of 25 Drosophila melanogaster host genotypes, naturally infected with Wolbachia and annotated for TE insertions. The host genotypes differed significantly in Wolbachia titers inside individual flies, with broad-sense heritability around 20%, and in the number of TE insertions, which depended greatly on TE identity. By removing Wolbachia from the target host genotypes, we generated a panel of 25 pairs of Wolbachia-positive and Wolbachia-negative lines in which we quantified transcription levels for our target TEs. We found variation in TE expression that was dependent on Wolbachia status, TE identity, and host genotype. Comparing between pairs of Wolbachia-positive and Wolbachia-negative flies, we found that Wolbachia removal affected TE expression in 21.1% of the TE-genotype combinations tested, with up to 2.3 times differences in the median level of transcript. Our data show that Wolbachia can impact TE activity in host genomes, underscoring the importance this endosymbiont can have in the generation of genetic novelty in hosts.}, } @article {pmid36928089, year = {2023}, author = {Terretaz, K and Horard, B and Weill, M and Loppin, B and Landmann, F}, title = {Functional analysis of Wolbachia Cid effectors unravels cooperative interactions to target host chromatin during replication.}, journal = {PLoS pathogens}, volume = {19}, number = {3}, pages = {e1011211}, pmid = {36928089}, issn = {1553-7374}, mesh = {Animals ; Male ; Chromatin/metabolism ; *Wolbachia/physiology ; Semen/metabolism ; Animals, Genetically Modified ; Drosophila/metabolism ; Cytoplasm/metabolism ; Centromere Protein A/metabolism ; *Drosophila Proteins/metabolism ; }, abstract = {Wolbachia are common bacteria among terrestrial arthropods. These endosymbionts transmitted through the female germline manipulate their host reproduction through several mechanisms whose most prevalent form called Cytoplasmic Incompatibility -CI- is a conditional sterility syndrome eventually favoring the infected progeny. Upon fertilization, the sperm derived from an infected male is only compatible with an egg harboring a compatible Wolbachia strain, this sperm leading otherwise to embryonic death. The Wolbachia Cif factors CidA and CidB responsible for CI and its neutralization function as a Toxin-Antitoxin system in the mosquito host Culex pipiens. However, the mechanism of CidB toxicity and its neutralization by the CidA antitoxin remain unexplored. Using transfected insect cell lines to perform a structure-function analysis of these effectors, we show that both CidA and CidB are chromatin interactors and CidA anchors CidB to the chromatin in a cell-cycle dependent-manner. In absence of CidA, the CidB toxin localizes to its own chromatin microenvironment and acts by preventing S-phase completion, independently of its deubiquitylase -DUB- domain. Experiments with transgenic Drosophila show that CidB DUB domain is required together with CidA during spermatogenesis to stabilize the CidA-CidB complex. Our study defines CidB functional regions and paves the way to elucidate the mechanism of its toxicity.}, } @article {pmid36914655, year = {2023}, author = {Kageyama, D and Harumoto, T and Nagamine, K and Fujiwara, A and Sugimoto, TN and Jouraku, A and Tamura, M and Katoh, TK and Watada, M}, title = {A male-killing gene encoded by a symbiotic virus of Drosophila.}, journal = {Nature communications}, volume = {14}, number = {1}, pages = {1357}, pmid = {36914655}, issn = {2041-1723}, mesh = {*Drosophila melanogaster/embryology/virology ; *Insect Viruses/genetics ; *Genes, Viral/physiology ; *Symbiosis ; Male ; Animals ; Embryonic Development ; RNA, Viral/physiology ; RNA, Double-Stranded/physiology ; Evolution, Molecular ; Open Reading Frames/genetics ; *Sex Ratio ; Sex Characteristics ; Female ; }, abstract = {In most eukaryotes, biparentally inherited nuclear genomes and maternally inherited cytoplasmic genomes have different evolutionary interests. Strongly female-biased sex ratios that are repeatedly observed in various arthropods often result from the male-specific lethality (male-killing) induced by maternally inherited symbiotic bacteria such as Spiroplasma and Wolbachia. However, despite some plausible case reports wherein viruses are raised as male-killers, it is not well understood how viruses, having much smaller genomes than bacteria, are capable of inducing male-killing. Here we show that a maternally inherited double-stranded RNA (dsRNA) virus belonging to the family Partitiviridae (designated DbMKPV1) induces male-killing in Drosophila. DbMKPV1 localizes in the cytoplasm and possesses only four genes, i.e., one gene in each of the four genomic segments (dsRNA1-dsRNA4), in contrast to ca. 1000 or more genes possessed by Spiroplasma or Wolbachia. We also show that a protein (designated PVMKp1; 330 amino acids in size), encoded by a gene on the dsRNA4 segment, is necessary and sufficient for inducing male-killing. Our results imply that male-killing genes can be easily acquired by symbiotic viruses through reassortment and that symbiotic viruses are hidden players in arthropod evolution. We anticipate that host-manipulating genes possessed by symbiotic viruses can be utilized for controlling arthropods.}, } @article {pmid36913994, year = {2023}, author = {Junsiri, W and Kamkong, P and Chinkangsadarn, T and Ouisuwan, S and Taweethavonsawat, P}, title = {Molecular identification and genetic diversity of equine ocular setariasis in Thailand based on the COI, 12S rDNA, and ITS1 regions.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {110}, number = {}, pages = {105425}, doi = {10.1016/j.meegid.2023.105425}, pmid = {36913994}, issn = {1567-7257}, mesh = {Animals ; Horses/genetics ; *Setariasis/diagnosis ; DNA, Ribosomal/genetics ; Thailand/epidemiology ; Phylogeny ; *Genetic Variation ; }, abstract = {Equine ocular setariasis is mainly caused by Setaria digitata, and the identification of this filarial nematode is based on morphology. However, morphological characterization alone is insufficient for the detection and differentiation of S. digitata from its congeners. In Thailand, the molecular detection of S. digitata is lacking and its genetic diversity is still unknown. This study aimed to phylogenetically characterize equine S. digitata from Thailand based on sequences derived from the mitochondrial cytochrome c oxidase subunit 1 (COI), the mitochondrial small subunit ribosomal DNA (12S rDNA), the nuclear internal transcribed spacer 1 (ITS1) and Wolbachia surface protein (wsp). Five samples of S. digitata were characterized, submitted to the NCBI database, and used for phylogenetic analysis as well as the assessment of similarity, entropy, and haplotype diversity. Phylogenetic analyses revealed that the S. digitata Thai strain was similar to S. digitata from China and Sri Lanka, with 99 to 100% similarity. The entropy and haplotype diversity indicated that the S. digitata Thai isolate was conserved and closely related to S. digitata worldwide. This is the first report on the molecular detection of equine ocular setariasis caused by S. digitata in Thailand.}, } @article {pmid36911919, year = {2023}, author = {Radousky, YA and Hague, MTJ and Fowler, S and Paneru, E and Codina, A and Rugamas, C and Hartzog, G and Cooper, BS and Sullivan, W}, title = {Distinct Wolbachia localization patterns in oocytes of diverse host species reveal multiple strategies of maternal transmission.}, journal = {Genetics}, volume = {224}, number = {1}, pages = {}, pmid = {36911919}, issn = {1943-2631}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; R35 GM139595/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Wolbachia/genetics ; Drosophila melanogaster ; Oocytes ; Oogenesis ; Drosophila/genetics ; }, abstract = {A broad array of endosymbionts radiate through host populations via vertical transmission, yet much remains unknown concerning the cellular basis, diversity, and routes underlying this transmission strategy. Here, we address these issues, by examining the cellular distributions of Wolbachia strains that diverged up to 50 million years ago in the oocytes of 18 divergent Drosophila species. This analysis revealed 3 Wolbachia distribution patterns: (1) a tight clustering at the posterior pole plasm (the site of germline formation); (2) a concentration at the posterior pole plasm, but with a significant bacteria population distributed throughout the oocyte; and (3) a distribution throughout the oocyte, with none or very few located at the posterior pole plasm. Examination of this latter class indicates Wolbachia accesses the posterior pole plasm during the interval between late oogenesis and the blastoderm formation. We also find that 1 Wolbachia strain in this class concentrates in the posterior somatic follicle cells that encompass the pole plasm of the developing oocyte. In contrast, strains in which Wolbachia concentrate at the posterior pole plasm generally exhibit no or few Wolbachia in the follicle cells associated with the pole plasm. Taken together, these studies suggest that for some Drosophila species, Wolbachia invade the germline from neighboring somatic follicle cells. Phylogenomic analysis indicates that closely related Wolbachia strains tend to exhibit similar patterns of posterior localization, suggesting that specific localization strategies are a function of Wolbachia-associated factors. Previous studies revealed that endosymbionts rely on 1 of 2 distinct routes of vertical transmission: continuous maintenance in the germline (germline-to-germline) or a more circuitous route via the soma (germline-to-soma-to-germline). Here, we provide compelling evidence that Wolbachia strains infecting Drosophila species maintain the diverse arrays of cellular mechanisms necessary for both of these distinct transmission routes. This characteristic may account for its ability to infect and spread globally through a vast range of host insect species.}, } @article {pmid36909520, year = {2023}, author = {Martin, M and Newton, ILG}, title = {The Wolbachia WalE1 effector alters Drosophila endocytosis.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36909520}, abstract = {The most common intracellular bacterial infection is Wolbachia pipientis, a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by Wolbachia have been studied for decades and range from sperm-egg incompatibility to male killing. How Wolbachia alters host biology is less well understood. Previously, we characterized the first Wolbachia effector - WalE1, which encodes a synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases Wolbachia titer in the developing oocyte. In this work, we first identify the native expression WalE1 by Wolbachia infecting both fly cells and whole animals. WalE1 appears as aggregates, separate from Wolbachia cells. We next show that WalE1 co-immunoprecipitates with the host protein Past1 and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in Past1 or overexpressing WalE1 are sensitive to AgNO3, a hallmark of endocytosis defects. Finally, we also show that Past1 null flies harbor more Wolbachia overall and in late egg chambers. Our results identify interactions between a Wolbachia secreted effector and a host protein and point to yet another important host cell process impinged upon by Wolbachia.}, } @article {pmid36907292, year = {2023}, author = {Ruiz, A and Gutiérrez-Bugallo, G and Rodríguez-Roche, R and Pérez, L and González-Broche, R and Piedra, LA and Martínez, LC and Menéndez, Z and Vega-Rúa, A and Bisset, JA}, title = {First report of natural Wolbachia infections in mosquitoes from Cuba.}, journal = {Acta tropica}, volume = {242}, number = {}, pages = {106891}, doi = {10.1016/j.actatropica.2023.106891}, pmid = {36907292}, issn = {1873-6254}, mesh = {Animals ; *Wolbachia/genetics ; Phylogeny ; Cuba ; Mosquito Vectors/microbiology ; *Aedes/microbiology ; }, abstract = {Mosquitoes are extensively responsible for the transmission of pathogens. Novel strategies using Wolbachia could transform that scenario, since these bacteria manipulate mosquito reproduction, and can confer a pathogen transmission-blocking phenotype in culicids. Here, we screened the Wolbachia surface protein region by PCR in eight Cuban mosquito species. We confirmed the natural infections by sequencing and assessed the phylogenetic relationships among the Wolbachia strains detected. We identified four Wolbachia hosts: Aedes albopictus, Culex quinquefasciatus, Mansonia titillans, and Aedes mediovittatus (first report worldwide). Knowledge of Wolbachia strains and their natural hosts is essential for future operationalization of this vector control strategy in Cuba.}, } @article {pmid36897930, year = {2023}, author = {Davison, HR and Hurst, GDD and Siozios, S}, title = {'Candidatus Megaira' are diverse symbionts of algae and ciliates with the potential for defensive symbiosis.}, journal = {Microbial genomics}, volume = {9}, number = {3}, pages = {}, pmid = {36897930}, issn = {2057-5858}, mesh = {*Symbiosis ; *Ciliophora ; Phylogeny ; }, abstract = {Symbiotic microbes from the genus 'Candidatus Megaira' (Rickettsiales) are known to be common associates of algae and ciliates. However, genomic resources for these bacteria are scarce, limiting our understanding of their diversity and biology. We therefore utilize Sequence Read Archive and metagenomic assemblies to explore the diversity of this genus. We successfully extract four draft 'Ca. Megaira' genomes including one complete scaffold for a 'Ca. Megaira' and identify an additional 14 draft genomes from uncategorized environmental metagenome-assembled genomes. We use this information to resolve the phylogeny for the hyper-diverse 'Ca. Megaira', with hosts broadly spanning ciliates, and micro- and macro-algae, and find that the current single genus designation 'Ca. Megaira' significantly underestimates their diversity. We also evaluate the metabolic potential and diversity of ''Ca. Megaira' from this new genomic data and find no clear evidence of nutritional symbiosis. In contrast, we hypothesize a potential for defensive symbiosis in 'Ca. Megaira'. Intriguingly, one symbiont genome revealed a proliferation of ORFs with ankyrin, tetratricopeptide and leucine-rich repeats such as those observed in the genus Wolbachia where they are considered important for host–symbiont protein–protein interactions. Onward research should investigate the phenotypic interactions between 'Ca. Megaira' and their various potential hosts, including the economically important Nemacystus decipiens, and target acquisition of genomic information to reflect the diversity of this massively variable group.}, } @article {pmid36889655, year = {2023}, author = {Zhu, X and Liu, T and He, A and Zhang, L and Li, J and Li, T and Miao, X and You, M and You, S}, title = {Diversity of Wolbachia infection and its influence on mitochondrial DNA variation in the diamondback moth, Plutella xylostella.}, journal = {Molecular phylogenetics and evolution}, volume = {182}, number = {}, pages = {107751}, doi = {10.1016/j.ympev.2023.107751}, pmid = {36889655}, issn = {1095-9513}, mesh = {Animals ; *Moths/genetics ; *Wolbachia/genetics ; Phylogeny ; DNA, Mitochondrial/genetics ; Mitochondria/genetics ; }, abstract = {Plutella xylostella is a pest that severely damages cruciferous vegetables worldwide and has been shown to be infected with the maternally inherited bacteria Wolbachia, with the main infected strain was plutWB1. In this study, we performed a large-scale global sampling of P. xylostella and amplified 3 mtDNA genes of P. xylostella and 6 Wolbachia genes to analyze the infection status, diversity of Wolbachia in P. xylostella, and its effect on mtDNA variation in P. xylostella. This study provides a conservative estimate of Wolbachia infection rates in P. xylostella, which was found to be 7% (104/1440). The ST 108 (plutWB1) was shared among butterfly species and the moth species P. xylostella, revealing that Wolbachia strain plutWB1 acquisition in P. xylostella may be through horizontal transmission. The Parafit analyses indicated a significant association between Wolbachia and Wolbachia-infected P. xylostella individuals, and individuals infected with plutWB1 tended to cluster in the basal positions of the phylogenetic tree based on the mtDNA data. Additionally, Wolbachia infections were associated with increased mtDNA polymorphism in the infected P. xylostella population. These data suggest that Wolbachia endosymbionts may have a potential effect on mtDNA variation of P. xylostella.}, } @article {pmid36889142, year = {2023}, author = {Hussain, M and Etebari, K and Asgari, S}, title = {Analysing inhibition of dengue virus in Wolbachia-infected mosquito cells following the removal of Wolbachia.}, journal = {Virology}, volume = {581}, number = {}, pages = {48-55}, doi = {10.1016/j.virol.2023.02.017}, pmid = {36889142}, issn = {1096-0341}, mesh = {Animals ; *Dengue Virus/physiology ; *Wolbachia/physiology ; Virus Replication ; *RNA Viruses/genetics ; *Aedes ; *Dengue ; }, abstract = {Wolbachia pipientis is known to block replication of positive sense RNA viruses. Previously, we created an Aedes aegypti Aag2 cell line (Aag2.wAlbB) transinfected with the wAlbB strain of Wolbachia and a matching tetracycline-cured Aag2.tet cell line. While dengue virus (DENV) was blocked in Aag2.wAlbB cells, we found significant inhibition of DENV in Aag2.tet cells. RNA-Seq analysis of the cells confirmed removal of Wolbachia and lack of expression of Wolbachia genes that could have been due to lateral gene transfer in Aag2.tet cells. However, we noticed a substantial increase in the abundance of phasi charoen-like virus (PCLV) in Aag2.tet cells. When RNAi was used to reduce the PCLV levels, DENV replication was significantly increased. Further, we found significant changes in the expression of antiviral and proviral genes in Aag2.tet cells. Overall, the results reveal an antagonistic interaction between DENV and PCLV and how PCLV-induced changes could contribute to DENV inhibition.}, } @article {pmid36883556, year = {2023}, author = {Bueno-Marí, R and Domínguez-Santos, R and Trelis, M and Garrote-Sánchez, E and Cholvi, M and Quero de Lera, F and Khoubbane, M and Marcilla, A and Gil, R}, title = {[Wolbachia pipientis infections in populations of Aedes albopictus in the city of València (Spain): implications for mosquito control].}, journal = {Revista espanola de salud publica}, volume = {97}, number = {}, pages = {}, pmid = {36883556}, issn = {2173-9110}, mesh = {Adult ; Male ; Animals ; Humans ; *Aedes ; Mosquito Control ; Spain/epidemiology ; *Wolbachia ; }, abstract = {OBJECTIVE: The presence of Aedes albopictus, of high sanitary and social impact, was first reported in Valencia (Eastern Spain) in 2015. Innovative tools for its control include the use of the endosymbiotic bacterium Wolbachia pipientis. The release of mosquito males infected with the wPip strain, has proven very promising for large-scale Incompatible Insect Technique (IIT) applications. Before this strategy can be implemented in Valencia, it is important to know whether the natural local mosquito populations are Wolbachia-infected and, if so, identifying the infecting strains/supergroups, these being the objectives of the present work.

METHODS: Eggs were collected from the 19 districts of the València city between May and October 2019. A total of 50 lab-reared adult Ae. albopictus individuals were processed and analyzed for Wolbachia detection and molecular characterization. These actions took place within the framework of a collaboration established with the Department of Health and Consumer Affairs of the city council of Valencia. Fisher's exact test was used to detect the statistical significance of the differences between groups.

RESULTS: Our study revealed that 94% of the analyzed samples were naturally infected with Wolbachia. Both wAlbA and wAlbB supergroups were identified, with most samples (72% of the infected ones) carrying co-infections.

CONCLUSIONS: These data provide the first characterization of the Wolbachia presence in natural populations of Ae. albopictus in the Mediterranean area of Spain. This information is relevant to evaluate the potential use of Wolbachia strains in order to achieve the suppression of the Asian tiger mosquito populations through massive release of artificially-infected males.}, } @article {pmid36877376, year = {2023}, author = {Zhang, Z and Chang, L and Huang, Q and Yan, R and Zheng, B}, title = {A mosquito population suppression model with a saturated Wolbachia release strategy in seasonal succession.}, journal = {Journal of mathematical biology}, volume = {86}, number = {4}, pages = {51}, pmid = {36877376}, issn = {1432-1416}, mesh = {Animals ; Female ; Male ; Seasons ; *Wolbachia ; *Culicidae ; Cytoplasm ; Cytosol ; }, abstract = {Releasing Wolbachia-infected male mosquitoes to suppress wild female mosquitoes through cytoplasmic incompatibility has shown great promise in controlling and preventing mosquito-borne diseases. To make the release logistically and economically feasible, we propose a saturated release strategy, which is only implemented during the epidemic season of mosquito-borne diseases. Under this assumption, the model becomes a seasonally switching ordinary differential equation model. The seasonal switch brings rich dynamics, including the existence of a unique periodic solution or exactly two periodic solutions, which are proved by using the qualitative property of the Poincaré map. Sufficient conditions are also obtained for determining the stability of the periodic solutions.}, } @article {pmid36869841, year = {2023}, author = {Cooper, WR and Walker, WB and Angelella, GM and Swisher Grimm, KD and Foutz, JJ and Harper, SJ and Nottingham, LB and Northfield, TD and Wohleb, CH and Strausbaugh, CA}, title = {Bacterial Endosymbionts Identified From Leafhopper (Hemiptera: Cicadellidae) Vectors of Phytoplasmas.}, journal = {Environmental entomology}, volume = {52}, number = {2}, pages = {243-253}, doi = {10.1093/ee/nvad015}, pmid = {36869841}, issn = {1938-2936}, mesh = {Male ; Animals ; *Hemiptera/genetics ; *Phytoplasma/genetics ; Bacteria/genetics ; Polymerase Chain Reaction ; Insect Vectors/microbiology ; Plant Diseases/microbiology ; }, abstract = {Insects often harbor bacterial endosymbionts that provide them with nutritional benefit or with protection against natural enemies, plant defenses, insecticides, and abiotic stresses. Certain endosymbionts may also alter acquisition and transmission of plant pathogens by insect vectors. We identified bacterial endosymbionts from four leafhopper vectors (Hemiptera: Cicadellidae) of 'Candidatus Phytoplasma' species by direct sequencing 16S rDNA and confirmed endosymbiont presence and identity by species-specific conventional PCR. We examined three vectors of Ca. Phytoplasma pruni, causal agent of cherry X-disease [Colladonus geminatus (Van Duzee), Colladonus montanus reductus (Van Duzee), Euscelidius variegatus (Kirschbaum)] - and a vector of Ca. Phytoplasma trifolii, the causal agent of potato purple top disease [Circulifer tenellus (Baker)]. Direct sequencing of 16S identified the two obligate endosymbionts of leafhoppers, 'Ca. Sulcia' and 'Ca. Nasuia', which are known to produce essential amino acids lacking in the leafhoppers' phloem sap diet. About 57% of C. geminatus also harbored endosymbiotic Rickettsia. We identified 'Ca. Yamatotoia cicadellidicola' in Euscelidius variegatus, providing just the second host record for this endosymbiont. Circulifer tenellus harbored the facultative endosymbiont Wolbachia, although the average infection rate was only 13% and all males were Wolbachia-uninfected. A significantly greater percentage of Wolbachia-infected Ci. tenellus adults than uninfected adults carried Ca. P. trifolii, suggesting that Wolbachia may increase this insect's ability to tolerate or acquire this pathogen. Results of our study provide a foundation for continued work on interactions between leafhoppers, bacterial endosymbionts, and phytoplasma.}, } @article {pmid36861014, year = {2023}, author = {Cheema, HA and Mujtaba, RS and Siddiqui, A and Vohra, LI and Shahid, A and Shah, J and Nashwan, AJ and Howard, N}, title = {Singapore's Dengue Outbreak Amidst the COVID-19 Pandemic: Challenges, Responses, and Lessons.}, journal = {Infection and drug resistance}, volume = {16}, number = {}, pages = {1081-1085}, pmid = {36861014}, issn = {1178-6973}, abstract = {Dengue outbreaks have been documented in Singapore since 1901, occurring almost annually in the 1960s and disproportionately affecting the paediatric population. In January 2020, virological surveillance detected a shift from DENV-2, which was the previous dominant strain, to DENV-3. As of 20 September 2022, 27,283 cases have been reported in 2022. Singapore is currently also responding to the COVID-19 pandemic, overcoming another wave of infections with 281,977 cases recorded in the past two months as of 19 September 2022. While Singapore has adopted several policies and interventions to combat dengue, primarily through environmental control but also innovations such as the Wolbachia mosquito programme, there is a need for further efforts to deal with the dual threat of dengue and COVID-19. Drawing lessons from Singapore's experience, countries facing such dual epidemics should enact clear policy responses, including establishing a multisectoral dengue action committee and action plan prior to potential outbreaks. Key indicators should be agreed upon and tracked at all healthcare levels as part of dengue surveillance and incorporated into the national health information system. Digitizing dengue monitoring systems and implementing telemedicine solutions are innovative measures that would facilitate the response to dengue in the context of restrictions during the COVID-19 pandemic that hinder the detection and response to new cases. There is a need for greater international collaboration in reducing or eradicating dengue in endemic countries. Further research is also required on how best to establish integrated early warning systems and extend our knowledge of the effects of COVID-19 on dengue transmission in affected countries.}, } @article {pmid36857313, year = {2023}, author = {Frantz, SI and Small, CM and Cresko, WA and Singh, ND}, title = {Ovarian transcriptional response to Wolbachia infection in D. melanogaster in the context of between-genotype variation in gene expression.}, journal = {G3 (Bethesda, Md.)}, volume = {13}, number = {5}, pages = {}, pmid = {36857313}, issn = {2160-1836}, mesh = {Animals ; *Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Genotype ; Gene Expression Profiling ; Transcriptome ; Symbiosis ; }, abstract = {Wolbachia is a maternally transmitted endosymbiotic bacteria that infects a wide variety of arthropod and nematode hosts. The effects of Wolbachia on host biology are far-reaching and include changes in host gene expression. However, previous work on the host transcriptional response has generally been investigated in the context of a single host genotype. Thus, the relative effect of Wolbachia infection versus vs. host genotype on gene expression is unknown. Here, we explicitly test the relative roles of Wolbachia infection and host genotype on host gene expression by comparing the ovarian transcriptomes of 4 strains of Drosophila melanogaster (D. melanogaster) infected and uninfected with Wolbachia. Our data suggest that infection explains a small amount of transcriptional variation, particularly in comparison to variation in gene expression among strains. However, infection specifically affects genes related to cell cycle, translation, and metabolism. We also find enrichment of cell division and recombination processes among genes with infection-associated differential expression. Broadly, the transcriptomic changes identified in this study provide novel understanding of the relative magnitude of the effect of Wolbachia infection on gene expression in the context of host genetic variation and also point to genes that are consistently differentially expressed in response to infection among multiple genotypes.}, } @article {pmid36850014, year = {2023}, author = {Latrofa, MS and Varotto-Boccazzi, I and Louzada-Flores, VN and Iatta, R and Mendoza-Roldan, JA and Roura, X and Zatelli, A and Epis, S and Bandi, C and Otranto, D}, title = {Interaction between Wolbachia pipientis and Leishmania infantum in heartworm infected dogs.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {77}, pmid = {36850014}, issn = {1756-3305}, mesh = {Female ; Animals ; Dogs ; *Dirofilaria immitis ; *Wolbachia ; *Leishmania infantum ; Cytokines ; }, abstract = {BACKGROUND: Wolbachia is a Gram-negative endosymbiont associated with several species of arthropods and filarioid nematodes, including Dirofilaria immitis. This endosymbiont may elicit a Th1 response, which is a component of the immunity against Leishmania infantum.

METHODS: To investigate the interactions between Wolbachia of D. immitis and L. infantum in naturally infected dogs and cytokine circulation, dogs without clinical signs (n = 187) were selected. Dogs were tested for microfilariae (mfs) by Knott, for female antigens of D. immitis by SNAP, and for anti-L. infantum antibodies by IFAT and assigned to four groups. Dogs of group 1 (G1) and 2 (G2) were positive for D. immitis and positive or negative to L. infantum, respectively. Dogs of group 3 (G3) and 4 (G4) were negative to D. immitis and positive or negative to L. infantum, respectively. Wolbachia and L. infantum DNA was quantified by real-time PCR (qPCR) in dog blood samples. A subset of dogs (n = 65) was examined to assess pro- and anti-inflammatory cytokine production using an ELISA test.

RESULTS: Of 93 dogs positive to D. immitis with circulating mfs, 85% were positive to Wolbachia, with the highest amount of DNA detected in G1 and the lowest in dogs with low mfs load in G1 and G2. Among dogs positive to L. infantum, 66% from G1 showed low antibody titer, while 48.9% from G3 had the highest antibody titer. Of 37 dogs positive to Wolbachia from G1, 26 (70.3%) had low antibody titers to L. infantum (1:160). Among cytokines, TNFα showed the highest mean concentration in G1 (246.5 pg/ml), IFNγ being the one most represented (64.3%). IL-10 (1809.5 pg/ml) and IL-6 (123.5 pg/ml) showed the highest mean concentration in dogs from G1. A lower percentage of dogs producing IL-4 was observed in all groups examined, with the highest mean concentration (2794 pg/ml) recorded in G2.

CONCLUSION: Results show the association of D. immitis and Wolbachia with the lower antibody titers of L. infantum in co-infected dogs, suggesting the hypothesis that the endosymbiont may affect the development of the patent leishmaniosis. However, due to the limitations associated with the heterogeneity of naturally infected dogs in field conditions, results should be validated by investigation on experimental models.}, } @article {pmid36844640, year = {2023}, author = {Algamdi, AG and Shaher, FM and Mahyoub, JA}, title = {Biological comparative study between Wolbachia-infected Aedes aegypti mosquito and Wolbachia-uninfected strain, Jeddah city, Saudi Arabia.}, journal = {Saudi journal of biological sciences}, volume = {30}, number = {3}, pages = {103581}, pmid = {36844640}, issn = {1319-562X}, abstract = {In this study, samples of Wolbachia-infected Aedes aegypti mosquitoes were collected from Al-Safa district in Jeddah city, Saudi Arabia. The presence of Wolbachia bacteria in mosquitoes was confirmed by PCR technique and they were reared and propagated in the laboratory. Comparative studies were conducted between Wolbachia-infected A. Aegypti and the Wolbachia-uninfected laboratory strain in terms of their ability to withstand drought, resist two types of insecticides and the activities of pesticide detoxification enzymes. The Wolbachia-infected A. aegypti strain proved less able to withstand the drought period, as the egg-hatching rate of the Wolbachia-uninfected strain was greater than that of the Wolbachia-infected strain after one, two and three months of dry periods. Compared to the Wolbachia-uninfected strain, the Wolbachia-infected strain demonstrated a relatively greater resistance to tested pesticides, namely Baton 100EC and Fendure 25EC which may be attributed to the higher levels of the detoxification enzymes glutathione-S-transferase and catalase and the lower levels of esterase and acetylcholine esterase.}, } @article {pmid36844098, year = {2023}, author = {Li, T and Yang, G and Li, Q and Jiang, Y and Kang, D and Fan, Z and Gong, Z and Lu, R and Zhou, G and Wu, Y and Lu, C}, title = {Population dynamics of migrant wheat aphids in China's main wheat production region and their interactions with bacterial symbionts.}, journal = {Frontiers in plant science}, volume = {14}, number = {}, pages = {1103236}, pmid = {36844098}, issn = {1664-462X}, abstract = {Sitobion miscanthi, Rhopalosiphum padi, and Schizaphis graminum are the three main pests in Chinese wheat-producing regions. In 2020, they are classified into the Chinese Class I list of agricultural diseases and pests, due to their severe harm to wheat plantings. S. miscanthi, R. padi, and S. graminum are migrant pests, and understanding their migration patterns and simulating their migration trajectories would improve forecasting and controlling them. Furthermore, the bacterial community of the migrant wheat aphid is also less known. In this study, we employed a suction trap to uncover the migration patterns of the three wheat aphid species in Yuanyang county, Henan province, during 2018 to 2020. And then the migration trajectories of S. miscanthi and R. padi were simulated using the NOAA HYSPLIT model. The interactions between wheat aphids and bacteria were further revealed by specific PCR and 16S rRNA amplicon sequencing. The results showed that the population dynamics of migrant wheat aphids was varied. Most of the trapped samples were identified to be R. padi, and S. graminum was the least collected sample. Typically, R. padi had two migration peaks in the 3 years, whereas S. miscanthi and S. graminum only exhibited one migration peak in 2018 and 2019. Moreover, the aphid migration trajectories varied over the years. Generally, the aphids originated from the south and migrated to the north. Herein, the infections of three main aphid facultative bacterial symbionts, Serratia symbiotica, Hamiltonella defensa, and Regiella insercticola, were detected in S. miscanthi and R. padi with specific PCR. Rickettsiella, Arsenophonus, Rickettsia, and Wolbachia were further identified with 16S rRNA amplicon sequencing. Biomarker searching indicated that Arsenophonus was significantly enriched in R. padi. Furthermore, diversity analyses showed that the bacterial community of R. padi had a higher richness and evenness than that of S. miscanthi. In conclusion, this study expands our knowledge about the migration patterns of aphids in the main wheat plant region of China and reveals the interactions between bacterial symbionts and migrant aphids.}, } @article {pmid36835753, year = {2023}, author = {Rodpai, R and Boonroumkaew, P and Sadaow, L and Sanpool, O and Janwan, P and Thanchomnang, T and Intapan, PM and Maleewong, W}, title = {Microbiome Composition and Microbial Community Structure in Mosquito Vectors Aedes aegypti and Aedes albopictus in Northeastern Thailand, a Dengue-Endemic Area.}, journal = {Insects}, volume = {14}, number = {2}, pages = {}, pmid = {36835753}, issn = {2075-4450}, support = {The Fundamental Fund of Khon Kaen University from the National Science, Research and Innovation Fund (NSRF)//Khon Kaen University/ ; }, abstract = {Bacterial content in mosquito larvae and adults is altered by dynamic interactions during life and varies substantially in variety and composition depending on mosquito biology and ecology. This study aimed to identify the microbiota in Aedes aegypti and Aedes albopictus and in water from their breeding sites in northeastern Thailand, a dengue-endemic area. Bacterial diversity in field-collected aquatic larvae and subsequently emerged adults of both species from several locations were examined. The microbiota was characterized based on analysis of DNA sequences from the V3-V4 region of the 16S rRNA gene and exhibited changes during development, from the mosquito larval stage to the adult stage. Aedes aegypti contained a significantly higher number of bacterial genera than did Ae. albopictus, except for the genus Wolbachia, which was present at significantly higher frequencies in male Ae. albopictus (p < 0.05). Our findings also indicate likely transstadial transmission from larva to adult and give better understanding of the microbial diversity in these mosquitoes, informing future control programs against mosquito-borne diseases.}, } @article {pmid36825089, year = {2023}, author = {Jiang, RX and Shang, F and Jiang, HB and Dou, W and Cernava, T and Wang, JJ}, title = {Candidatus Liberibacter asiaticus: An important factor affecting bacterial community composition and Wolbachia titers in Asian citrus psyllid.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1109803}, pmid = {36825089}, issn = {1664-302X}, abstract = {Endosymbionts play crucial roles in various physiological activities within insect hosts. The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is an important vector for Candidatus Liberibacter asiaticus (CLas), a fatal pathogenic bacterial agent causing the disease Huanglongbing in the citrus industry. This study combines high-throughput sequencing of 16S ribosomal RNA amplicons to explore how CLas affects the bacterial community in different color morphs (blue, gray), genders, and tissues (cuticle, gut, mycetome, Malpighian tubule, ovary, and testis) of ACP. We found that there was no significant differences in the bacterial community diversity and CLas acquired ratio between the different color morphs and genders of ACP adults. However, acquiring CLas could promote the adult bacterial community's diversity and richness more than in the uninfected condition. The presence of CLas could increase the Wolbachia and unclassified_Enterobacteriaceae proportions more than in the uninfected condition. The bacterial community diversity in the CLas infected tissues of ovary and cuticle, was lower than the uninfected condition, but the richness of all tissues was not different between the infected and uninfected conditions. CLas could also change the bacterial structure in different tissues and make the bacterial relationship network simpler than it is in an uninfected condition. Furthermore, we used quantitative real-time PCR to assess the dynamic changes of Wolbachia in CLas uninfected and infected color morphs and tissues of ACP. The results showed that Wolbachia titers were significantly higher in CLas infected adults than in uninfected adults. In different tissues, the Wolbachia titers in the testis, ovary, and Malpighian tubule were higher than their uninfected counterparts. Our results provide essential knowledge for understanding the symbionts of the ACP and how CLas affects the bacterial community of the ACP.}, } @article {pmid36819029, year = {2023}, author = {Liu, B and Ren, YS and Su, CY and Abe, Y and Zhu, DH}, title = {Pangenomic analysis of Wolbachia provides insight into the evolution of host adaptation and cytoplasmic incompatibility factor genes.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1084839}, pmid = {36819029}, issn = {1664-302X}, abstract = {INTRODUCTION: The genus Wolbachia provides a typical example of intracellular bacteria that infect the germline of arthropods and filarial nematodes worldwide. Their importance as biological regulators of invertebrates, so it is particularly important to study the evolution, divergence and host adaptation of these bacteria at the genome-wide level.

METHODS: Here, we used publicly available Wolbachia genomes to reconstruct their evolutionary history and explore their adaptation under host selection.

RESULTS: Our findings indicate that segmental and single-gene duplications, such as DNA methylase, bZIP transcription factor, heat shock protein 90, in single monophyletic Wolbachia lineages (including supergroups A and B) may be responsible for improving the ability to adapt to a broad host range in arthropod-infecting strains. In contrast to A strains, high genetic diversity and rapidly evolving gene families occur in B strains, which may promote the ability of supergroup B strains to adapt to new hosts and their large-scale spreading. In addition, we hypothesize that there might have been two independent horizontal transfer events of cif genes in two sublineages of supergroup A strains. Interestingly, during the independent evolution of supergroup A and B strains, the rapid evolution of cif genes in supergroup B strains resulted in the loss of their functional domain, reflected in a possible decrease in the proportion of induced cytoplasmic incompatibility (CI) strains.

DISCUSSION: This present study highlights for reconstructing of evolutionary history, addressing host adaptation-related evolution and exploring the origin and divergence of CI genes in each Wolbachia supergroup. Our results thus not only provide a basis for further exploring the evolutionary history of Wolbachia adaptation under host selection but also reveal a new research direction for studying the molecular regulation of Wolbachia- induced cytoplasmic incompatibility.}, } @article {pmid36817770, year = {2023}, author = {Fischer, K and Dubben, B and Debrah, LB and Kuehlwein, JM and Ricchiuto, A and Debrah, AY and Hoerauf, A and Weil, GJ and Fischer, PU and Klarmann-Schulz, U}, title = {Histopathological evaluation of Onchocerca volvulus nodules by microscopy and by digital image analysis for the study of macrofilaricidal drug efficacy.}, journal = {Frontiers in medicine}, volume = {10}, number = {}, pages = {1099926}, pmid = {36817770}, issn = {2296-858X}, abstract = {BACKGROUND: Novel drugs or drug combinations that kill or permanently sterilize adult Onchocerca volvulus worms would be very helpful for treatment and elimination of onchocerciasis. In absence of a reliable biomarker for viable adult worms, histopathological assessment of worms within onchocercal nodules is a standard method to determine macrofilaricidal activity. The goal of the present study was to determine the agreement between two independent experts in the analysis of nodule sections and to assess the value of digital imaging as a means of standardizing the analysis.

MATERIAL AND METHODS: Two expert microscopists independently assessed 605 nodules by direct microscopy. At least two sections with two different stains hematoxylin & eosin (H&E, APR immunostain) of paraffin-embedded, ethanol-fixed whole-nodule cross-sections were analyzed. After variables were identified prone to observer discrepancies, we performed a second study to compare consolidated results for 100 nodules obtained by the two readers by microscopy and by analysis of scanned, high resolution digital images (20x magnification). The last data set analyzed was a quality panel of 100 nodules that has been previously examined by microscopy, and included additional immunostains for Wolbachia endobacteria. These slides were digitalized, read by the two assessors and results were compared with original microscopy results.

RESULTS: The degree of agreement between assessors varied for different parameters. Agreement for female worm counts in nodules was approximately 80%, while agreement regarding female worm viability was 98%. There were no major differences observed between results obtained by microscopy or digital images. Good agreement for important parameters was also observed for the nodules of the quality panel.

CONCLUSION: Nodule analysis by experienced microscopists was reproducible with regard to important parameters such as identification of living female worms or detection of normal embryogenesis. Assessments varied more for other parameters, and we recommend continued use of two independent readers for detailed analyzes. Analysis of scanned images provided similar results to direct microscopy. This facilitates training and comparison of nodule findings by readers in different locations. Analysis of high quality digital images that can be viewed remotely should improve the quality and availability of nodule assessments that are primary endpoints for onchocerciasis clinical trials.}, } @article {pmid36810669, year = {2023}, author = {Manoj, RRS and Latrofa, MS and Louni, M and Laidoudi, Y and Fenollar, F and Otranto, D and Mediannikov, O}, title = {In vitro maintenance of the endosymbiont Wolbachia of Dirofilaria immitis.}, journal = {Parasitology research}, volume = {122}, number = {4}, pages = {939-943}, pmid = {36810669}, issn = {1432-1955}, mesh = {Animals ; Dogs ; *Dirofilaria immitis ; *Dirofilariasis ; *Wolbachia ; Microfilariae ; *Dog Diseases/microbiology ; }, abstract = {Wolbachia has an obligatory mutualistic relationship with many onchocercid nematodes of the subfamilies Dirofilariinae and Onchocercinae. Till date, no attempts have been made for the in vitro cultivation of this intracellular bacterium from the filarioid host. Hence, the current study attempted cell co-culture method using embryonic Drosophila S2 and the LD cell lines to cultivate Wolbachia from Dirofilaria immitis microfilariae (mfs) harvested from infected dogs. Microfilariae (mfs = 1500) were inoculated in shell vials supplemented with Schneider medium using both cell lines. The establishment and multiplication of the bacterium were observed during the initial inoculation, at day 0 and before every medium change (from days 14 to 115). An aliquot (50 µl) from each time point was tested by quantitative real-time PCR (qPCR). Comparing the average of Ct values, obtained by the tested parameters (i.e., LD/S2 cell lines and mfs with/without treatment), the S2 cell line without mechanical disruption of mfs provided the highest Wolbachia cell count by qPCR. Despite the maintenance of Wolbachia within both S2 and LD-based cell co-culture models for up to 115 days, a definitive conclusion is still far. Further trials using fluorescent microscopy and viable staining will help to demonstrate the cell line infection and viability of Wolbachia. Use of considerable amount of untreated mfs to inoculate the Drosophilia S2 cell lines, as well as the supplementation of the culture media with growth stimulants or pre-treated cells to increase their susceptibility for the infection and development of a filarioid-based cell line system are recommended for the future trials.}, } @article {pmid36810610, year = {2023}, author = {Muro, T and Hikida, H and Fujii, T and Kiuchi, T and Katsuma, S}, title = {Two Complete Genomes of Male-Killing Wolbachia Infecting Ostrinia Moth Species Illuminate Their Evolutionary Dynamics and Association with Hosts.}, journal = {Microbial ecology}, volume = {86}, number = {3}, pages = {1740-1754}, pmid = {36810610}, issn = {1432-184X}, support = {17H06431//Japan Society for the Promotion of Science/ ; 22H00366//Japan Society for the Promotion of Science/ ; }, mesh = {Animals ; Male ; *Moths/genetics ; *Wolbachia/genetics ; Phylogeny ; Sex Ratio ; Genomics ; }, abstract = {Wolbachia is an extremely widespread intracellular symbiont which causes reproductive manipulation on various arthropod hosts. Male progenies are killed in Wolbachia-infected lineages of the Japanese Ostrinia moth population. While the mechanism of male killing and the evolutionary interaction between host and symbiont are significant concerns for this system, the absence of Wolbachia genomic information has limited approaches to these issues. We determined the complete genome sequences of wFur and wSca, the male-killing Wolbachia of Ostrinia furnacalis and Ostrinia scapulalis. The two genomes shared an extremely high degree of homology, with over 95% of the predicted protein sequences being identical. A comparison of these two genomes revealed nearly minimal genome evolution, with a strong emphasis on the frequent genome rearrangements and the rapid evolution of ankyrin repeat-containing proteins. Additionally, we determined the mitochondrial genomes of both species' infected lineages and performed phylogenetic analyses to deduce the evolutionary dynamics of Wolbachia infection in the Ostrinia clade. According to the inferred phylogenetic relationship, two possible scenarios were proposed: (1) Wolbachia infection was established in the Ostrinia clade prior to the speciation of related species such as O. furnacalis and O. scapulalis, or (2) Wolbachia infection in these species was introgressively transferred from a currently unidentified relative. Simultaneously, the relatively high homology of mitochondrial genomes suggested recent Wolbachia introgression between infected Ostrinia species. The findings of this study collectively shed light on the host-symbiont interaction from an evolutionary standpoint.}, } @article {pmid36800397, year = {2023}, author = {Mills, MK and McCabe, LG and Rodrigue, EM and Lechtreck, KF and Starai, VJ}, title = {Wbm0076, a candidate effector protein of the Wolbachia endosymbiont of Brugia malayi, disrupts eukaryotic actin dynamics.}, journal = {PLoS pathogens}, volume = {19}, number = {2}, pages = {e1010777}, pmid = {36800397}, issn = {1553-7374}, mesh = {Animals ; Humans ; Actins/metabolism ; *Brugia malayi/genetics ; Eukaryotic Cells ; Saccharomyces cerevisiae/genetics ; Symbiosis/genetics ; *Wolbachia/physiology ; Bacterial Proteins ; }, abstract = {Brugia malayi, a parasitic roundworm of humans, is colonized by the obligate intracellular bacterium, Wolbachia pipientis. The symbiosis between this nematode and bacterium is essential for nematode reproduction and long-term survival in a human host. Therefore, identifying molecular mechanisms required by Wolbachia to persist in and colonize B. malayi tissues will provide new essential information regarding the basic biology of this endosymbiosis. Wolbachia utilize a Type IV secretion system to translocate so-called "effector" proteins into the cytosol of B. malayi cells to promote colonization of the eukaryotic host. However, the characterization of these Wolbachia secreted proteins has remained elusive due to the genetic intractability of both organisms. Strikingly, expression of the candidate Wolbachia Type IV-secreted effector protein, Wbm0076, in the surrogate eukaryotic cell model, Saccharomyces cerevisiae, resulted in the disruption of the yeast actin cytoskeleton and inhibition of endocytosis. Genetic analyses show that Wbm0076 is a member of the family of Wiskott-Aldrich syndrome proteins (WAS [p]), a well-conserved eukaryotic protein family required for the organization of actin skeletal structures. Thus, Wbm0076 likely plays a central role in the active cell-to-cell movement of Wolbachia throughout B. malayi tissues during nematode development. As most Wolbachia isolates sequenced to date encode at least partial orthologs of wBm0076, we find it likely that the ability of Wolbachia to directly manipulate host actin dynamics is an essential requirement of all Wolbachia endosymbioses, independent of host cell species.}, } @article {pmid36786616, year = {2023}, author = {Li, C and Liu, S and Zhou, H and Zhu, W and Cui, M and Li, J and Wang, J and Liu, J and Zhu, J and Li, W and Bi, Y and Carr, MJ and Holmes, EC and Shi, W}, title = {Metatranscriptomic Sequencing Reveals Host Species as an Important Factor Shaping the Mosquito Virome.}, journal = {Microbiology spectrum}, volume = {11}, number = {2}, pages = {e0465522}, pmid = {36786616}, issn = {2165-0497}, abstract = {Mosquitoes are important vector hosts for numerous viral pathogens and harbor a large number of mosquito-specific viruses as well as human-infecting viruses. Previous studies have mainly focused on the discovery of mosquito viruses, and our understanding of major ecological factors associated with virome structure in mosquitoes remains limited. We utilized metatranscriptomic sequencing to characterize the viromes of five mosquito species sampled across eight locations in Yunnan Province, China. This revealed the presence of 52 viral species, of which 19 were novel, belonging to 15 viral families/clades. Of particular note was Culex hepacivirus 1, clustering within the avian clade of hepaciviruses. Notably, both the viromic diversity and abundance of Aedes genus mosquitoes were significantly higher than those of the Culex genus, while Aedes albopictus mosquitoes harbored a higher diversity than Aedes aegypti mosquitoes. Our findings thus point to discernible differences in viromic structure between mosquito genera and even between mosquito species within the same genus. Importantly, such differences were not attributable to differences in sampling between geographical location. Our study also revealed the ubiquitous presence of the endosymbiont bacterium Wolbachia, with the genetic diversity and abundance also varying between mosquito species. In conclusion, our results suggested that the mosquito host species play an important role in shaping the virome's structure. IMPORTANCE This study revealed the huge capability of mosquitoes in harboring a rich diversity of RNA viruses, although relevant studies have characterized the intensively unparalleled diversity of RNA viruses previously. Furthermore, our findings showed discernible differences not only in viromic structure between mosquito genera and even between mosquito species within the same genus but also in the genetic diversity and abundance of Wolbachia between different mosquito populations. These findings emphasize the importance of host genetic background in shaping the virome composition of mosquitoes.}, } @article {pmid36785954, year = {2023}, author = {Sanaei, E and Albery, GF and Yeoh, YK and Lin, YP and Cook, LG and Engelstädter, J}, title = {Host phylogeny and ecological associations best explain Wolbachia host shifts in scale insects.}, journal = {Molecular ecology}, volume = {32}, number = {9}, pages = {2351-2363}, doi = {10.1111/mec.16883}, pmid = {36785954}, issn = {1365-294X}, mesh = {Animals ; *Hemiptera/microbiology ; Insecta/genetics ; Phylogeny ; Symbiosis/genetics ; Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are among the most prevalent and widespread endosymbiotic bacteria on Earth. Wolbachia's success in infecting an enormous number of arthropod species is attributed to two features: the range of phenotypes they induce in their hosts, and their ability to switch between host species. Whilst much progress has been made in elucidating their induced phenotypes, our understanding of Wolbachia host-shifting is still very limited: we lack answers to even fundamental questions concerning Wolbachia's routes of transfer and the importance of factors influencing host shifts. Here, we investigate the diversity and host-shift patterns of Wolbachia in scale insects, a group of arthropods with intimate associations with other insects that make them well suited to studying host shifts. Using Illumina multitarget amplicon sequencing of Wolbachia-infected scale insects and their direct associates we determined the identity of all Wolbachia strains. We then fitted a generalized additive mixed model to our data to estimate the influence of host phylogeny and the geographical distribution on Wolbachia strain sharing among scale insect species. The model predicts no significant contribution of host geography but strong effects of host phylogeny, with high rates of Wolbachia sharing among closely related species and a sudden drop-off in sharing with increasing phylogenetic distance. We also detected the same Wolbachia strain in scale insects and several intimately associated species (ants, wasps and flies). This indicates putative host shifts and potential routes of transfers via these associates and highlights the importance of ecological connectivity in Wolbachia host-shifting.}, } @article {pmid36781724, year = {2023}, author = {Takasuka, K and Arakawa, K}, title = {The Method of Eliminating the Wolbachia Endosymbiont Genomes from Insect Samples Prior to a Long-Read Sequencing.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2632}, number = {}, pages = {101-112}, pmid = {36781724}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Wasps/genetics ; Genome ; Insecta/genetics ; Rifampin ; Symbiosis/genetics ; }, abstract = {When extracting DNA of invertebrates for long-read sequencing, not only enough quantity and size of the DNA but, depending on the species, elimination of contamination of endosymbiotic Wolbachia genome also has to be achieved. These requirements become troublesome, especially in small-sized species with a limited number of individuals available for the experiment. In this chapter, using tiny parasitoid wasps (Reclinervellus nielseni) parasitizing spiders as hosts, we developed a method of eliminating the Wolbachia genomes by means of an antibiotic administration to adult wasps via honey solution. Twenty days of rifampicin treatment since their emergence from cocoons resulted in a significant decrease in the Wolbachia genomes while keeping good DNA conditions for nanopore sequencing. An adequate quantity of DNA was then gained by pooling several individuals. The method could be applied to other insects or invertebrates that can be maintained by laboratory feeding with liquid food.}, } @article {pmid36778977, year = {2022}, author = {Sinha, DK and Gupta, A and Padmakumari, AP and Bentur, JS and Nair, S}, title = {Infestation of Rice by Gall Midge Influences Density and Diversity of Pseudomonas and Wolbachia in the Host Plant Microbiome.}, journal = {Current genomics}, volume = {23}, number = {2}, pages = {126-136}, pmid = {36778977}, issn = {1389-2029}, abstract = {Background: The virulence of phytophagous insects is predominantly determined by their ability to evade or suppress host defense for their survival. The rice gall midge (GM, Orseolia oryzae), a monophagous pest of rice, elicits a host defense similar to the one elicited upon pathogen attack. This could be due to the GM feeding behaviour, wherein the GM endosymbionts are transferred to the host plant via oral secretions, and as a result, the host mounts an appropriate defense response(s) (i.e., up-regulation of the salicylic acid pathway) against these endosymbionts. Methods: The current study aimed to analyze the microbiome present at the feeding site of GM maggots to determine the exchange of bacterial species between GM and its host and to elucidate their role in rice-GM interaction using a next-generation sequencing approach. Results: Our results revealed differential representation of the phylum Proteobacteria in the GM-infested and -uninfested rice tissues. Furthermore, analysis of the species diversity of Pseudomonas and Wolbachia supergroups at the feeding sites indicated the exchange of bacterial species between GM and its host upon infestation. Conclusion: As rice-GM microbial associations remain relatively unstudied, these findings not only add to our current understanding of microbe-assisted insect-plant interactions but also provide valuable insights into how these bacteria drive insect-plant coevolution. Moreover, to the best of our knowledge, this is the first report analyzing the microbiome of a host plant (rice) at the feeding site of its insect pest (GM).}, } @article {pmid36761111, year = {2022}, author = {Pang, Y and Su, CY and Zhu, JQ and Yang, XH and Zhong, JL and Zhu, DH and Liu, Z}, title = {A new species of Andricus Hartig, 1840 (Hymenoptera, Cynipidae) from China, with references to DNA taxonomy and Wolbachia infection.}, journal = {ZooKeys}, volume = {1134}, number = {}, pages = {52-73}, pmid = {36761111}, issn = {1313-2989}, abstract = {In the present paper, a new species of cynipid gall wasp, Andricuselodeoides Liu & Pang, is described from several provinces in southern China. The new species is closely related to the recently redescribed A.mairei (Kieffer, 1906). In addition to differences in adult and gall morphology, the new species is also readily separated by COI sequences, with a 6.2-8.9% genetic distance between populations of the new species and those of A.mairei. A contrasting difference in sex ratios was also observed between the two species, with A.elodeoides extremely female-biased (95.5-97.8% female) while A.mairei male-biased to more balanced (5.4-43.5% female). PCR screening for Wolbachia infection further revealed contrasting infection rates between populations of A.elodeoides and A.mairei: the Wolbachia infection rate was 0% in A.elodeoides and 100% in A.mairei. Cytoplasmic incompatibility induced by Wolbachia is proposed as a potential mechanism of speciation of the sympatric A.elodeoides and A.mairei.}, } @article {pmid36760975, year = {2022}, author = {Zarka, J and Parmentier, T and Wybouw, N}, title = {Intersexuality in a natural population of the terrestrial isopod Porcellioscaber.}, journal = {ZooKeys}, volume = {1101}, number = {}, pages = {183-190}, pmid = {36760975}, issn = {1313-2989}, abstract = {Intersex phenotypes are rarely observed in natural isopod populations and their expression is typically associated with infection of Wolbachia, a reproductive parasite that manipulates arthropod reproduction. During an intensive sampling effort of a natural population of the isopod Porcellioscaber, an adult individual was isolated that expressed both male and female traits. The intersex individual exhibited clearly developed external male genitalia and carried multiple eggs in its brood pouch. No Wolbachia infection could be identified in this individual, a result that needs to be approached with caution due to suboptimal DNA preservation for diagnostic PCR assays. Wolbachia were, however, detected in two adult females of the same population, and appear closely related to isolates that infect other terrestrial isopod species. This is the first demonstration that intersex phenotypes can arise under natural conditions in P.scaber.}, } @article {pmid36757767, year = {2023}, author = {Halter, T and Köstlbacher, S and Rattei, T and Hendrickx, F and Manzano-Marín, A and Horn, M}, title = {One to host them all: genomics of the diverse bacterial endosymbionts of the spider Oedothorax gibbosus.}, journal = {Microbial genomics}, volume = {9}, number = {2}, pages = {}, pmid = {36757767}, issn = {2057-5858}, mesh = {Animals ; *Spiders/genetics/microbiology ; DNA Transposable Elements ; Bacteroidetes/genetics ; Genomics ; *Rickettsia/genetics ; *Wolbachia/genetics ; }, abstract = {Bacterial endosymbionts of the groups Wolbachia, Cardinium and Rickettsiaceae are well known for their diverse effects on their arthropod hosts, ranging from mutualistic relationships to reproductive phenotypes. Here, we analysed a unique system in which the dwarf spider Oedothorax gibbosus is co-infected with up to five different endosymbionts affiliated with Wolbachia, 'Candidatus Tisiphia' (formerly Torix group Rickettsia), Cardinium and Rhabdochlamydia. Using short-read genome sequencing data, we show that the endosymbionts are heterogeneously distributed among O. gibbosus populations and are frequently found co-infecting spider individuals. To study this intricate host-endosymbiont system on a genome-resolved level, we used long-read sequencing to reconstruct closed genomes of the Wolbachia, 'Ca. Tisiphia' and Cardinium endosymbionts. We provide insights into the ecology and evolution of the endosymbionts and shed light on the interactions with their spider host. We detected high quantities of transposable elements in all endosymbiont genomes and provide evidence that ancestors of the Cardinium, 'Ca. Tisiphia' and Wolbachia endosymbionts have co-infected the same hosts in the past. Our findings contribute to broadening our knowledge about endosymbionts infecting one of the largest animal phyla on Earth and show the usefulness of transposable elements as an evolutionary 'contact-tracing' tool.}, } @article {pmid36754115, year = {2023}, author = {Prigot-Maurice, C and Lheraud, B and Guéritault, S and Beltran-Bech, S and Cordaux, R and Peccoud, J and Braquart-Varnier, C}, title = {Investigating Wolbachia symbiont-mediated host protection against a bacterial pathogen using a natural Wolbachia nuclear insert.}, journal = {Journal of invertebrate pathology}, volume = {197}, number = {}, pages = {107893}, doi = {10.1016/j.jip.2023.107893}, pmid = {36754115}, issn = {1096-0805}, mesh = {Female ; Animals ; *Symbiosis ; *Wolbachia ; Bacteria ; }, abstract = {Wolbachia bacterial endosymbionts provide protection against pathogens in various arthropod species but the underlying mechanisms remain misunderstood. By using a natural Wolbachia nuclear insert (f-element) in the isopod Armadillidium vulgare, we explored whether Wolbachia presence is mandatory to observe protection in this species or the presence of its genes is sufficient. We assessed survival of closely related females carrying or lacking the f-element (and lacking Wolbachia) challenged with the bacterial pathogen Salmonella enterica. Despite marginal significant effects, the f-element alone did not appear to confer survival benefits to its host, suggesting that Wolbachia presence in cells is crucial for protection.}, } @article {pmid36750860, year = {2023}, author = {Mahmood, S and Nováková, E and Martinů, J and Sychra, O and Hypša, V}, title = {Supergroup F Wolbachia with extremely reduced genome: transition to obligate insect symbionts.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {22}, pmid = {36750860}, issn = {2049-2618}, mesh = {Animals ; Phylogeny ; *Wolbachia/genetics ; Biological Evolution ; *Nematoda ; Insecta ; Symbiosis/physiology ; }, abstract = {BACKGROUND: Wolbachia belong to highly abundant bacteria which are frequently found in invertebrate microbiomes and manifest by a broad spectrum of lifestyles from parasitism to mutualism. Wolbachia supergroup F is a particularly interesting clade as it gave rise to symbionts of both arthropods and nematodes, and some of its members are obligate mutualists. Investigations on evolutionary transitions among the different symbiotic stages have been hampered by a lack of the known diversity and genomic data for the supergroup F members.

RESULTS: Based on amplicon screening, short- and long-read WGS approaches, and laser confocal microscopy, we characterize five new supergroup F Wolbachia strains from four chewing lice species. These strains reached different evolutionary stages and represent two remarkably different types of symbiont genomes. Three of the genomes resemble other known members of Wolbachia F supergroup, while the other two show typical signs of ongoing gene inactivation and removal (genome size, coding density, low number of pseudogenes). Particularly, wMeur1, a symbiont fixed in microbiomes of Menacanthus eurysternus across four continents, possesses a highly reduced genome of 733,850 bp. The horizontally acquired capacity for pantothenate synthesis and localization in specialized bacteriocytes suggest its obligate nutritional role.

CONCLUSIONS: The genome of wMeur1 strain, from the M. eurysternus microbiome, represents the smallest currently known Wolbachia genome and the first example of Wolbachia which has completed genomic streamlining as known from the typical obligate symbionts. This points out that despite the large amount and great diversity of the known Wolbachia strains, evolutionary potential of these bacteria still remains underexplored. The diversity of the four chewing lice microbiomes indicates that this vast parasitic group may provide suitable models for further investigations. Video Abstract.}, } @article {pmid36748430, year = {2022}, author = {Izraeli, Y and Lepetit, D and Atias, S and Mozes-Daube, N and Wodowski, G and Lachman, O and Luria, N and Steinberg, S and Varaldi, J and Zchori-Fein, E and Chiel, E}, title = {Genomic characterization of viruses associated with the parasitoid Anagyrus vladimiri (Hymenoptera: Encyrtidae).}, journal = {The Journal of general virology}, volume = {103}, number = {12}, pages = {}, doi = {10.1099/jgv.0.001810}, pmid = {36748430}, issn = {1465-2099}, mesh = {Humans ; Female ; Animals ; Phylogeny ; *Wasps ; *Viruses ; Genomics ; *Reoviridae/genetics ; }, abstract = {Knowledge on symbiotic microorganisms of insects has increased dramatically in recent years, yet relatively little data are available regarding non-pathogenic viruses. Here we studied the virome of the parasitoid wasp Anagyrus vladimiri Triapitsyn (Hymenoptera: Encyrtidae), a biocontrol agent of mealybugs. By high-throughput sequencing of viral nucleic acids, we revealed three novel viruses, belonging to the families Reoviridae [provisionally termed AnvRV (Anagyrus vladimiri reovirus)], Iflaviridae (AnvIFV) and Dicistroviridae (AnvDV). Phylogenetic analysis further classified AnvRV in the genus Idnoreovirus, and AnvDV in the genus Triatovirus. The genome of AnvRV comprises 10 distinct genomic segments ranging in length from 1.5 to 4.2 kb, but only two out of the 10 ORFs have a known function. AnvIFV and AnvDV each have one polypeptide ORF, which is typical of iflaviruses but very un-common among dicistroviruses. Five conserved domains were found along both the ORFs of those two viruses. AnvRV was found to be fixed in an A. vladimiri population that was obtained from a mass rearing facility, whereas its prevalence in field-collected A. vladimiri was ~15 %. Similarly, the prevalence of AnvIFV and AnvDV was much higher in the mass rearing population than in the field population. The presence of AnvDV was positively correlated with the presence of Wolbachia in the same individuals. Transmission electron micrographs of females' ovaries revealed clusters and viroplasms of reovirus-like particles in follicle cells, suggesting that AnvRV is vertically transmitted from mother to offspring. AnvRV was not detected in the mealybugs, supporting the assumption that this virus is truly associated with the wasps. The possible effects of these viruses on A. vladimiri's biology, and on biocontrol agents in general, are discussed. Our findings identify RNA viruses as potentially involved in the multitrophic system of mealybugs, their parasitoids and other members of the holobiont.}, } @article {pmid36744754, year = {2023}, author = {Chen, J and Wang, MK and Xie, QX and Bing, XL and Li, TP and Hong, XY}, title = {NDUFA8 potentially rescues Wolbachia-induced cytoplasmic incompatibility in Laodelphax striatellus.}, journal = {Insect science}, volume = {30}, number = {6}, pages = {1689-1700}, doi = {10.1111/1744-7917.13182}, pmid = {36744754}, issn = {1744-7917}, support = {32020103011//National Natural Science Foundation of China/ ; 32001905//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; }, mesh = {Female ; Male ; Animals ; *Wolbachia/genetics ; *Hemiptera/genetics/metabolism ; Fertility ; Reproduction ; Gene Expression Profiling ; }, abstract = {The endosymbiont Wolbachia manipulates host reproduction by several strategies, one of the most important of which is cytoplasmic incompatibility (CI). CI can be rescued when Wolbachia-infected males mate with females infected with the same Wolbachia strain. However, the potential rescue mechanism of CI in the small brown planthopper Laodelphax striatellus is unclear. In this study, comparative transcriptome analysis was applied to explore the effect of Wolbachia on L. striatellus eggs. A total of 1387 differentially expressed genes were identified. RNA interference of 7 Wolbachia-upregulated key planthopper genes reduced egg reproduction, suggesting that Wolbachia might improve fecundity in L. striatellus by affecting these 7 genes. Suppressing the expression of another upregulated gene, NDUFA8 (encoding NADH dehydrogenase [ubiquinone] 1 α subcomplex subunit 8-like) by RNA interference significantly increased the mortality of early embryos without affecting the number of deposited eggs. Wolbachia infection upregulated the mRNA level of NDUFA8, and dsNDUFA8 treatment of Wolbachia-infected females recreated CI-like symptoms, suggesting that NDUFA8 is associated with the rescue phenotype. Because all L. striatellus populations worldwide are infected with Wolbachia, NDUFA8 is a potential pest control target.}, } @article {pmid36719928, year = {2023}, author = {Ross, PA and Elfekih, S and Collier, S and Klein, MJ and Lee, SS and Dunn, M and Jackson, S and Zhang, Y and Axford, JK and Gu, X and Home, JL and Nassar, MS and Paradkar, PN and Tawfik, EA and Jiggins, FM and Almalik, AM and Al-Fageeh, MB and Hoffmann, AA}, title = {Developing Wolbachia-based disease interventions for an extreme environment.}, journal = {PLoS pathogens}, volume = {19}, number = {1}, pages = {e1011117}, pmid = {36719928}, issn = {1553-7374}, mesh = {Animals ; *Wolbachia ; Saudi Arabia ; Australia ; Extreme Environments ; *Aedes ; *Dengue ; }, abstract = {Aedes aegypti mosquitoes carrying self-spreading, virus-blocking Wolbachia bacteria are being deployed to suppress dengue transmission. However, there are challenges in applying this technology in extreme environments. We introduced two Wolbachia strains into Ae. aegypti from Saudi Arabia for a release program in the hot coastal city of Jeddah. Wolbachia reduced infection and dissemination of dengue virus (DENV2) in Saudi Arabian mosquitoes and showed complete maternal transmission and cytoplasmic incompatibility. Wolbachia reduced egg hatch under a range of environmental conditions, with the Wolbachia strains showing differential thermal stability. Wolbachia effects were similar across mosquito genetic backgrounds but we found evidence of local adaptation, with Saudi Arabian mosquitoes having lower egg viability but higher adult desiccation tolerance than Australian mosquitoes. Genetic background effects will influence Wolbachia invasion dynamics, reinforcing the need to use local genotypes for mosquito release programs, particularly in extreme environments like Jeddah. Our comprehensive characterization of Wolbachia strains provides a foundation for Wolbachia-based disease interventions in harsh climates.}, } @article {pmid36715911, year = {2023}, author = {Nevalainen, LBM and Newton, ILG}, title = {Detection and Assessment of Wolbachia pipientis Infection.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2626}, number = {}, pages = {291-307}, pmid = {36715911}, issn = {1940-6029}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Multilocus Sequence Typing ; In Situ Hybridization, Fluorescence ; Drosophila/genetics ; }, abstract = {Wolbachia pipientis is a widespread vertically transmitted intracellular bacterium naturally present in the model organism Drosophila melanogaster. As Wolbachia is present in a large number of Drosophila lines, it is critical for researchers to be able to identify which of their stocks maintain this infection to avoid any potential confounding variables. Here, we describe methods for detecting the bacterium and assessing the infection, including polymerase chain reaction (PCR) of DNA, multi-locus sequence typing (MLST) to identify strains, western blotting for protein detection, and immunohistochemistry and fluorescence in situ hybridization (FISH) of Drosophila ovaries to visually detect Wolbachia by fluorescence microscopy.}, } @article {pmid36714832, year = {2022}, author = {Liang, X and Tan, CH and Sun, Q and Zhang, M and Wong, PSJ and Li, MI and Mak, KW and Martín-Park, A and Contreras-Perera, Y and Puerta-Guardo, H and Manrique-Saide, P and Ng, LC and Xi, Z}, title = {Wolbachia wAlbB remains stable in Aedes aegypti over 15 years but exhibits genetic background-dependent variation in virus blocking.}, journal = {PNAS nexus}, volume = {1}, number = {4}, pages = {pgac203}, pmid = {36714832}, issn = {2752-6542}, abstract = {The ability of the maternally transmitted endosymbiotic bacterium Wolbachia to induce cytoplasmic incompatibility (CI) and virus blocking makes it a promising weapon for combatting mosquito-borne diseases through either suppression or replacement of wild-type populations. Recent field trials show that both approaches significantly reduce the incidence of dengue fever in humans. However, new questions emerge about how Wolbachia-mosquito associations will co-evolve over time and whether Wolbachia-mediated virus blocking will be affected by the genetic diversity of mosquitoes and arboviruses in the real world. Here, we have compared the Wolbachia density and CI expression of two wAlbB-infected Aedes aegypti lines transinfected 15 years apart. We have also assessed wAlbB-mediated virus blocking against dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) viruses and examined whether host genetic backgrounds modulate viral blocking effects by comparing ZIKV infection in mosquitoes with a Mexican genetic background to those with a Singaporean background. Our results show that over 15 years, wAlbB maintained the capacity to form a stable association with Ae. aegypti in terms of both density and CI expression. There were variations in wAlbB-induced virus blocking against CHIKV, DENV, and ZIKV, and higher inhibitory effects on ZIKV in mosquitoes on the Singaporean genetic background than on the Mexican background. These results provide important information concerning the robustness and long-term stability of Wolbachia as a biocontrol agent for arbovirus disease control.}, } @article {pmid36714306, year = {2022}, author = {Barman, M and Samanta, S and Ahmed, B and Dey, S and Chakraborty, S and Deeksha, MG and Dutta, S and Samanta, A and Tarafdar, J and Roy, D}, title = {Transcription dynamics of heat-shock proteins (Hsps) and endosymbiont titres in response to thermal stress in whitefly, Bemisia tabaci (Asia-I).}, journal = {Frontiers in physiology}, volume = {13}, number = {}, pages = {1097459}, pmid = {36714306}, issn = {1664-042X}, abstract = {The sweet potato whitefly, Bemisia tabaci (Gennadius), is one of the several species complexes of whitefly that are currently significant agricultural pests. Bemisia tabaci infests more than 600 plant species and thrives under a wide range of temperature conditions. In addition to the direct damage caused by sucking plant sap, it vectors several plant viruses. Heat-shock proteins play a pivotal role in enabling the insect to extend its geographical location, survival, and reproduction under different stress conditions. B. tabaci harbours several endosymbionts under the genera Portiera, Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium, and Fritschea that directly or indirectly affect its fitness. By accelerating cuticle biosynthesis and sclerotisation, symbiotic microbes can reduce or enhance tolerance to extreme temperatures and detoxify heavy metals. Thus, symbionts or microbial communities can expand or constrain the abiotic niche space of their host and affect its ability to adapt to changing conditions. The present study delineates the effect of thermal stress on the expression of heat-shock genes and endosymbionts in B. tabaci. Studies of the expression level of heat-shock proteins with the help of quantitative real-time polymerase chain reaction (qRT-PCR) showed that heat- and cold-shock treatment fuels the increased expression of heat-shock proteins (Hsp40 and Hsp70). However, Hsp90 was not induced by a heat- and cold-shock treatment. A significant decrease in the relative titre of secondary endosymbionts, such as Rickettsia, Arsenophonus, and Wolbachia, were recorded in B. tabaci upon heat treatment. However, the titre of the primary symbiont, C. Portiera, was relatively unaffected by both cold and heat treatments. These results are indicative of the fact that Hsp genes and endosymbionts in B. tabaci are modulated in response to thermal stress, and this might be responsible for the adaptation of whitefly under changing climatic scenario.}, } @article {pmid36712932, year = {2023}, author = {Herran, B and Sugimoto, TN and Watanabe, K and Imanishi, S and Tsuchida, T and Matsuo, T and Ishikawa, Y and Kageyama, D}, title = {Cell-based analysis reveals that sex-determining gene signals in Ostrinia are pivotally changed by male-killing Wolbachia.}, journal = {PNAS nexus}, volume = {2}, number = {1}, pages = {pgac293}, pmid = {36712932}, issn = {2752-6542}, abstract = {Wolbachia, a maternally transmitted bacterium, shows male-killing, an adaptive phenotype for cytoplasmic elements, in various arthropod species during the early developmental stages. In lepidopteran insects, lethality of males is accounted for by improper dosage compensation in sex-linked genes owing to Wolbachia-induced feminization. Herein, we established Ostrinia scapulalis cell lines that retained sex specificity per the splicing pattern of the sex-determining gene doublesex (Osdsx). We found that Wolbachia transinfection in male cell lines enhanced the female-specific splice variant of Osdsx (Osdsx[F]) while suppressing the male-specific variant (Osdsx[M]), indicating that Wolbachia affects sex-determining gene signals even in vitro. Comparative transcriptome analysis isolated only two genes that behave differently upon Wolbachia infection. The two genes were respectively homologous to Masculinizer (BmMasc) and zinc finger-2 (Bmznf-2), male-specifically expressed sex-determining genes of the silkworm Bombyx mori that encode CCCH-type zinc finger motif proteins. By using cultured cells and organismal samples, OsMasc and Osznf-2 were found to be sex-determining genes of O. scapulalis that are subjected to sex-specific alternative splicing depending upon the chromosomal sex, developmental stage, and infection status. Overall, our findings expound the cellular autonomy in insect sex determination and the mechanism through which sex is manipulated by intracellular selfish microbes.}, } @article {pmid36711723, year = {2023}, author = {Dodson, BL and Pujhari, S and Brustolin, M and Metz, HC and Rasgon, JL}, title = {Variable effects of Wolbachia on alphavirus infection in Aedes aegypti.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36711723}, abstract = {Wolbachia pipientis (=Wolbachia) has promise as a tool to suppress virus transmission by Aedes aegypti mosquitoes. However, Wolbachia can have variable effects on mosquito-borne viruses. This variation remains poorly characterized, yet the multimodal effects of Wolbachia on diverse pathogens could have important implications for public health. Here, we examine the effects of somatic infection with two strains of Wolbachia (wAlbB and wMel) on the alphaviruses Sindbis virus (SINV), O'nyong-nyong virus (ONNV), and Mayaro virus (MAYV) in Ae. aegypti. We found variable effects of Wolbachia including enhancement and suppression of viral infections, with some effects depending on Wolbachia strain. Both wAlbB- and wMel-infected mosquitoes showed enhancement of SINV infection rates one week post-infection, with wAlbB-infected mosquitoes also having higher viral titers than controls. Infection rates with ONNV were low across all treatments and no significant effects of Wolbachia were observed. The effects of Wolbachia on MAYV infections were strikingly strain-specific; wMel strongly blocked MAYV infections and suppressed viral titers, while wAlbB did not influence MAYV infection. The variable effects of Wolbachia on vector competence underscore the importance of further research into how this bacterium impacts the virome of wild mosquitoes including the emergent human pathogens they transmit.}, } @article {pmid36711506, year = {2023}, author = {Lindsey, AR and Parish, AJ and Newton, IL and Tennessen, JM and Jones, MW and Stark, N}, title = {Wolbachia is a nutritional symbiont in Drosophila melanogaster.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36711506}, abstract = {The intracellular bacterium Wolbachia is a common symbiont of many arthropods and nematodes, well studied for its impacts on host reproductive biology. However, its broad success as a vertically transmitted infection cannot be attributed to manipulations of host reproduction alone. Using the Drosophila melanogaster model and their natively associated Wolbachia strain "wMel", we show that Wolbachia infection supports fly development and buffers against nutritional stress. Wolbachia infection across several fly genotypes and a range of nutrient conditions resulted in reduced pupal mortality, increased adult emergence, and larger size. We determined that the exogenous supplementation of pyrimidines rescued these phenotypes in the Wolbachia-free, flies suggesting that Wolbachia plays a role in providing this metabolite that is normally limiting for fly growth. Additionally, Wolbachia was sensitive to host pyrimidine metabolism: Wolbachia titers increased upon transgenic knockdown of the Drosophila de novo pyrimidine synthesis pathway but not knockdown of the de novo purine synthesis pathway. We propose that Wolbachia acts as a nutritional symbiont to supplement fly development and enhance host fitness.}, } @article {pmid36708161, year = {2023}, author = {Hubert, J and Nesvorna, M and Sopko, B and Green, SJ}, title = {Diet modulation of the microbiome of the pest storage mite Tyrophagus putrescentiae.}, journal = {FEMS microbiology ecology}, volume = {99}, number = {3}, pages = {}, doi = {10.1093/femsec/fiad011}, pmid = {36708161}, issn = {1574-6941}, mesh = {Animals ; *Acaridae/genetics/microbiology ; RNA, Ribosomal, 16S/genetics ; *Microbiota ; *Mites ; Diet ; Bacteria/genetics ; Bacteroidetes/genetics ; Enterobacteriaceae/genetics ; }, abstract = {Storage mites colonize a wide spectrum of food commodities and adaptations to diets have been suggested as mechanisms enabling successful colonization. We characterized the response of seven unique Tyrophagus putrescentiae cultures (5K, 5L, 5N, 5P, 5Pi, 5S, and 5Tk) with different baseline microbiomes to different diets. The offered diets included a rearing diet, protein-enriched diet, oat flakes, and sunflower seeds. Microbiome characterization was performed using 16S ribosomal RNA (rRNA) gene amplicon sequencing and 16S rRNA gene quantitative PCR. The mite culture microbiomes were classified into four groups: (i) Sodalis-dominated (5Pi), (ii) Wolbachia-dominated (5N and 5P), (iii) Cardinium-dominated (5L and 5S), and (iv) asymbiontic (5K and 5Tk) mites dominated by Bacillus and Bartonella. Mite growth rates were most strongly affected by nutrients in the diet, while respiration and microbial community profiles were largely influenced by mite culture. While growth rate was not directly explained by microbiome composition, microbiomes strongly influenced mite fitness as measured by respiration. While diet significantly influenced microbial profiles in all cultures, the effect of diet differed in impact between cultures (5Pi > 5S > 5N > 5K > 5Tk > 5L > 5P). Furthermore, no new bacterial taxa were acquired by mites after dietary changes. Bacteria from the taxa Bacillus, Bartonella-like, Solitalea-like, Kocuria, and Sodalis-like contributed most strongly to differentiating mite-associated microbiomes.}, } @article {pmid36706527, year = {2023}, author = {Wei, X and Peng, H and Li, Y and Meng, B and Wang, S and Bi, S and Zhao, X}, title = {Pyrethroids exposure alters the community and function of the internal microbiota in Aedes albopictus.}, journal = {Ecotoxicology and environmental safety}, volume = {252}, number = {}, pages = {114579}, doi = {10.1016/j.ecoenv.2023.114579}, pmid = {36706527}, issn = {1090-2414}, mesh = {Animals ; *Insecticides/pharmacology ; Permethrin/toxicity ; *Aedes ; Mosquito Vectors ; *Pyrethrins/pharmacology ; Insecticide Resistance/genetics ; *Microbiota ; }, abstract = {Large amounts of insecticides bring selection pressure and then develop insecticide resistance in Aedes albopictus. This study demonstrated for the first time the effect of pyrethroid exposure on the internal microbiota in Ae. albopictus. 36, 48, 57 strains of virgin adult Ae. albopictus were exposed to the pyrethroids deltamethrin (Dme group), β-cypermethrin (Bcy group), and cis-permethrin (Cper group), respectively, with n-hexane exposure (Hex group) as the controls (n = 36). The internal microbiota community and functions were analyzed based on the metagenomic analysis. The analysis of similarity (ANOSIM) results showed that the Hex/Bcy (p = 0.001), Hex/Cper (p = 0.006), Hex/Dme (p = 0.001) groups were well separated, and the internal microbes of Ae. albopictus vary in the composition and functions depending on the type of pyrethroid insecticide they are applied. Four short chain fatty acid-producing genera, Butyricimonas, Prevotellaceae, Anaerococcus, Pseudorhodobacter were specifically absent in the pyrethroid-exposed mosquitoes. Morganella and Streptomyces were significantly enriched in cis-permethrin-exposed mosquitoes. Wolbachia and Chryseobacterium showed significant enrichment in β-cypermethrin-exposed mosquitoes. Pseudomonas was significantly abundant in deltamethrin-exposed mosquitoes. The significant proliferation of these bacteria may be closely related to insecticide metabolism. Our study recapitulated a specifically enhanced metabolic networks relevant to the exposure to cis-permethrin and β-cypermethrin, respectively. Benzaldehyde dehydrogenase (EC 1.2.1.28), key enzyme in aromatic compounds metabolism, was detected enhanced in cis-permethrin and β-cypermethrin exposed mosquitoes. The internal microbiota metabolism of aromatic compounds may be important influencing factors for pyrethroid resistance. Future work will be needed to elucidate the specific mechanisms by which mosquito microbiota influences host resistance and vector ability.}, } @article {pmid36700745, year = {2023}, author = {Indriani, C and Tanamas, SK and Khasanah, U and Ansari, MR and Rubangi, and Tantowijoyo, W and Ahmad, RA and Dufault, SM and Jewell, NP and Utarini, A and Simmons, CP and Anders, KL}, title = {Impact of randomised wmel Wolbachia deployments on notified dengue cases and insecticide fogging for dengue control in Yogyakarta City.}, journal = {Global health action}, volume = {16}, number = {1}, pages = {2166650}, pmid = {36700745}, issn = {1654-9880}, support = {R01 AI148127/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Humans ; *Aedes ; *Dengue/epidemiology/prevention & control ; *Dengue Virus ; *Insecticides ; *Wolbachia ; }, abstract = {BACKGROUND: Releases of Wolbachia (wMel)-infected Aedes aegypti mosquitoes significantly reduced the incidence of virologically confirmed dengue in a previous cluster randomised trial in Yogyakarta City, Indonesia. Following the trial, wMel releases were extended to the untreated control areas, to achieve city-wide coverage of Wolbachia.

OBJECTIVE: In this predefined analysis, we evaluated the impact of the wMel deployments in Yogyakarta on dengue hemorrhagic fever (DHF) case notifications and on the frequency of perifocal insecticide spraying by public health teams.

METHODS: Monthly counts of DHF cases notified to the Yogyakarta District Health Office between January 2006 and May 2022 were modelled as a function of time-varying local wMel treatment status (fully- and partially-treated vs untreated, and by quintile of wMel prevalence). The frequency of insecticide fogging in wMel-treated and untreated areas was analysed using negative binomial regression.

RESULTS: Notified DHF incidence was 83% lower in fully treated vs untreated periods (IRR 0.17 [95% CI 0.14, 0.20]), and 78% lower in areas with 80-100% wMel prevalence compared to areas with 0-20% wMel (IRR 0.23 [0.17, 0.30]). A similar intervention effect was observed at 60-80% wMel prevalence as at 80-100% prevalence (76% vs 78% efficacy, respectively). Pre-intervention, insecticide fogging occurred at similar frequencies in areas later randomised to wMel-treated and untreated arms of the trial. After wMel deployment, fogging occurred significantly less frequently in treated areas (IRR 0.17 [0.10, 0.30]).

CONCLUSIONS: Deployments of wMel-infected Aedes aegypti mosquitoes resulted in an 83% reduction in the application of perifocal insecticide spraying, consistent with lower dengue case notifications in wMel-treated areas. These results show that the Wolbachia intervention effect demonstrated previously in a cluster randomised trial was also measurable from routine surveillance data.}, } @article {pmid36691279, year = {2023}, author = {Cooper, WR and Swisher Grimm, KD and Angelella, GM and Mustafa, T}, title = {Acquisition and Transmission of 'Candidatus Liberibacter solanacearum' Differs Among Wolbachia-Infected and -Uninfected Haplotypes of Bactericera cockerelli.}, journal = {Plant disease}, volume = {107}, number = {8}, pages = {2440-2445}, doi = {10.1094/PDIS-11-22-2701-RE}, pmid = {36691279}, issn = {0191-2917}, mesh = {Animals ; Liberibacter ; Haplotypes ; Plant Diseases/microbiology ; *Solanum tuberosum/microbiology ; *Hemiptera/microbiology ; }, abstract = {'Candidatus Liberibacter solanacearum' (Lso) causes disease symptoms and economic losses in potato, tomato, and other solanaceous crops in North America. Lso is transmitted to plants by the potato psyllid, Bactericera cockerelli, which occurs as distinct haplotypes named western, central, and northwestern that differ in the presence or absence of the bacterial endosymbiont, Wolbachia. Previous work showed that all three vector haplotypes can transmit Lso, but it was not clear whether acquisition and transmission rates of Lso were equal among the haplotypes. The goal of our study was to compare Lso infection rates among psyllids of the western, central, and northwestern haplotypes. Using data collected from several years of periodic testing of Lso infection of laboratory-reared potato psyllid colonies, we showed that psyllids of the western and central haplotypes are more likely to harbor Lso than are psyllids of the northwestern haplotype. We then used greenhouse assays to demonstrate that psyllids of the northwestern haplotype are less likely to acquire and transmit Lso than those of the western haplotype. Lso infection rates corresponded with Wolbachia infection among the three psyllid haplotypes. The Wolbachia-infected central and western haplotypes were more likely to harbor and transmit Lso than the Wolbachia-free northwestern haplotype. Results demonstrate that potato psyllids of the western and central haplotypes pose a greater risk for spread of Lso in crops and suggest a pattern between infection with Lso and Wolbachia in potato psyllid.}, } @article {pmid36689552, year = {2023}, author = {Vancaester, E and Blaxter, M}, title = {Phylogenomic analysis of Wolbachia genomes from the Darwin Tree of Life biodiversity genomics project.}, journal = {PLoS biology}, volume = {21}, number = {1}, pages = {e3001972}, pmid = {36689552}, issn = {1545-7885}, support = {206194//Wellcome Trust/United Kingdom ; }, mesh = {Humans ; Animals ; Phylogeny ; *Wolbachia/genetics ; Genomics ; *Nematoda ; *Diptera ; Symbiosis/genetics ; }, abstract = {The Darwin Tree of Life (DToL) project aims to sequence all described terrestrial and aquatic eukaryotic species found in Britain and Ireland. Reference genome sequences are generated from single individuals for each target species. In addition to the target genome, sequenced samples often contain genetic material from microbiomes, endosymbionts, parasites, and other cobionts. Wolbachia endosymbiotic bacteria are found in a diversity of terrestrial arthropods and nematodes, with supergroups A and B the most common in insects. We identified and assembled 110 complete Wolbachia genomes from 93 host species spanning 92 families by filtering data from 368 insect species generated by the DToL project. From 15 infected species, we assembled more than one Wolbachia genome, including cases where individuals carried simultaneous supergroup A and B infections. Different insect orders had distinct patterns of infection, with Lepidopteran hosts mostly infected with supergroup B, while infections in Diptera and Hymenoptera were dominated by A-type Wolbachia. Other than these large-scale order-level associations, host and Wolbachia phylogenies revealed no (or very limited) cophylogeny. This points to the occurrence of frequent host switching events, including between insect orders, in the evolutionary history of the Wolbachia pandemic. While supergroup A and B genomes had distinct GC% and GC skew, and B genomes had a larger core gene set and tended to be longer, it was the abundance of copies of bacteriophage WO who was a strong determinant of Wolbachia genome size. Mining raw genome data generated for reference genome assemblies is a robust way of identifying and analysing cobiont genomes and giving greater ecological context for their hosts.}, } @article {pmid36689491, year = {2023}, author = {Li, J and Champer, J}, title = {Harnessing Wolbachia cytoplasmic incompatibility alleles for confined gene drive: A modeling study.}, journal = {PLoS genetics}, volume = {19}, number = {1}, pages = {e1010591}, pmid = {36689491}, issn = {1553-7404}, mesh = {Female ; Male ; Alleles ; Cytoplasm/genetics/microbiology ; Cytosol ; *Gene Drive Technology ; Reproduction/genetics ; Symbiosis ; *Wolbachia/genetics ; Animals ; *Symbiont Induced Cytoplasmic Incompatibility ; Genome, Insect ; Insecta/genetics/microbiology ; Transgenes ; }, abstract = {Wolbachia are maternally-inherited bacteria, which can spread rapidly in populations by manipulating reproduction. cifA and cifB are genes found in Wolbachia phage that are responsible for cytoplasmic incompatibility, the most common type of Wolbachia reproductive interference. In this phenomenon, no viable offspring are produced when a male with both cifA and cifB (or just cifB in some systems) mates with a female lacking cifA. Utilizing this feature, we propose new types of toxin-antidote gene drives that can be constructed with only these two genes in an insect genome, instead of the whole Wolbachia bacteria. By using both mathematical and simulation models, we found that a drive containing cifA and cifB together creates a confined drive with a moderate to high introduction threshold. When introduced separately, they act as a self-limiting drive. We observed that the performance of these drives is substantially influenced by various ecological parameters and drive characteristics. Extending our models to continuous space, we found that the drive individual release distribution has a critical impact on drive persistence. Our results suggest that these new types of drives based on Wolbachia transgenes are safe and flexible candidates for genetic modification of populations.}, } @article {pmid36683703, year = {2022}, author = {Liu, Y and He, ZQ and Wen, Q and Peng, J and Zhou, YT and Mandour, N and McKenzie, CL and Ahmed, MZ and Qiu, BL}, title = {Parasitoid-mediated horizontal transmission of Rickettsia between whiteflies.}, journal = {Frontiers in cellular and infection microbiology}, volume = {12}, number = {}, pages = {1077494}, pmid = {36683703}, issn = {2235-2988}, mesh = {Animals ; Female ; Phylogeny ; *Hemiptera/microbiology ; *Rickettsia/genetics ; In Situ Hybridization, Fluorescence ; Infectious Disease Transmission, Vertical ; Symbiosis ; }, abstract = {Intracellular bacterial endosymbionts of arthropods are mainly transmitted vertically from mother to offspring, but phylogenetically distant insect hosts often harbor identical endosymbionts, indicating that horizontal transmission from one species to another occurs in nature. Here, we investigated the parasitoid Encarsia formosa-mediated horizontal transmission of the endosymbiont Rickettsia between different populations of whitefly Bemisia tabaci MEAM1. Rickettsia was successfully transmitted from the positive MEAM1 nymphs (R [+]) into E. formosa and retained at least for 48 h in E. formosa adults. Fluorescence in situ hybridization (FISH) visualization results revealed that the ovipositors, mouthparts, and digestive tract of parasitoid adults get contaminated with Rickettsia. Random non-lethal probing of Rickettisia-negative (R[-]) MEAM1 nymphs by these Rickettsia-carrying E. formosa resulted in newly infected MEAM1 nymphs, and the vertical transmission of Rickettsia within the recipient females can remain at least up to F3 generation. Further phylogenetic analyses revealed that Rickettsia had high fidelity during the horizontal transmission in whiteflies and parasitoids. Our findings may help to explain why Rickettsia bacteria are so abundant in arthropods and suggest that, in some insect species that shared the same parasitoids, Rickettsia may be maintained in populations by horizontal transmission.}, } @article {pmid36680294, year = {2023}, author = {Ogunlade, ST and Meehan, MT and Adekunle, AI and McBryde, ES}, title = {A Systematic Review of Mathematical Models of Dengue Transmission and Vector Control: 2010-2020.}, journal = {Viruses}, volume = {15}, number = {1}, pages = {}, pmid = {36680294}, issn = {1999-4915}, mesh = {Animals ; Humans ; *Dengue/epidemiology/prevention & control ; *Aedes ; Mosquito Vectors ; Models, Theoretical ; *Wolbachia ; }, abstract = {Vector control methods are considered effective in averting dengue transmission. However, several factors may modify their impact. Of these controls, chemical methods, in the long run, may increase mosquitoes' resistance to chemicides, thereby decreasing control efficacy. The biological methods, which may be self-sustaining and very effective, could be hampered by seasonality or heatwaves (resulting in, e.g., loss of Wolbachia infection). The environmental methods that could be more effective than the chemical methods are under-investigated. In this study, a systematic review is conducted to explore the present understanding of the effectiveness of vector control approaches via dengue transmission models.}, } @article {pmid36677449, year = {2023}, author = {Hoffmann, A and Müller, T and Fingerle, V and Silaghi, C and Noll, M}, title = {Co-Infection of Potential Tick-Borne Pathogens of the Order Rickettsiales and Borrelia burgdorferi s. l. and Their Link to Season and Area in Germany.}, journal = {Microorganisms}, volume = {11}, number = {1}, pages = {}, pmid = {36677449}, issn = {2076-2607}, abstract = {The prevalence of potential human pathogenic members of the order Rickettsiales differs between Borrelia burgdorferi sensu lato-positive and -negative tick microbiomes. Here, co-infection of members of the order Rickettsiales, such as Rickettsia spp., Anaplasma phagocytophilum, Wolbachia pipientis, and Neoehrlichia mikurensis as well as B. burgdorferi s.l. in the tick microbiome was addressed. This study used conventional PCRs to investigate the diversity and prevalence of the before-mentioned bacteria in 760 nucleic acid extracts of I. ricinus ticks detached from humans, which were previously tested for B. burgdorferi s.l.. A gltA gene-based amplicon sequencing approach was performed to identify Rickettsia species. The prevalence of Rickettsia spp. (16.7%, n = 127) and W. pipientis (15.9%, n = 121) were similar, while A. phagocytophilum was found in 2.8% (n = 21) and N. mikurensis in 0.1% (n = 1) of all ticks. Co-infection of B. burgdorferi s. l. with Rickettsia spp. was most frequent. The gltA gene sequencing indicated that Rickettsia helvetica was the dominant Rickettsia species in tick microbiomes. Moreover, R, monacensis and R. raoultii were correlated with autumn and area south, respectively, and a negative B. burgdorferi s. l. finding. Almost every fifth tick carried DNA of at least two of the human pathogenic bacteria studied here.}, } @article {pmid36670494, year = {2023}, author = {Chamankar, B and Maleki-Ravasan, N and Karami, M and Forouzan, E and Karimian, F and Naeimi, S and Choobdar, N}, title = {The structure and diversity of microbial communities in Paederus fuscipes (Coleoptera: Staphylinidae): from ecological paradigm to pathobiome.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {11}, pmid = {36670494}, issn = {2049-2618}, mesh = {Humans ; Animals ; Male ; Female ; *Coleoptera/microbiology ; *Rhizobiaceae ; Enterococcus ; *Microbiota/genetics ; *Dermatitis ; }, abstract = {BACKGROUND: Paederus fuscipes is medically the most famous rove beetle, which causes dermatitis or conjunctivitis in humans, as well as gastrointestinal toxicosis in livestock, via releasing toxic hemolymph containing pederin. Pedrin biosynthesis genes have been identified in uncultured Pseudomonas-like endosymbionts that are speculated to be acquired through a horizontal transfer. However, the composition of the P. fuscipes microbial community, especially of the gut and genital microbiome, remains unclear. This study was aimed to characterize the structure and diversity of P. fuscipes-associated bacterial communities in terms of gender, organ, and location using the Illumina HiSeq platform in the southern littorals of Caspian Sea.

RESULTS: The OTUs identified from P. fuscipes specimens were collapsed into 40 phyla, 112 classes, 249 orders, 365 families, 576 genera, and 106 species. The most abundant families were Pseudomonadaceae, Spiroplasmataceae, Weeksellaceae, Enterococcaceae, and Rhizobiaceae, respectively. Thirty top genera made up > 94% of the P. fuscipes microbiome, with predominating Pseudomonas, followed by the Spiroplasma, Apibacter, Enterococcus, Dysgonomonas, Sebaldella, Ruminococcus, and Wolbachia. Interesting dissimilarities were also discovered within and between the beetle microbiomes in terms of genders and organs. Analyses showed that Spiroplasma / Apibacter as well as Pseudomonas / Pseudomonas were the most abundant in the genitals / intestines of male and female beetles, respectively. Bacterial richness did not display any significant difference in the three provinces but was higher in male beetles than in females and more in the genitals than intestines.

CONCLUSIONS: The present study identified Pseudomonas-like endobacterium as a common symbiont of P. fuscipes beetles; this bacterium begins its journey from gut and genitalia of females to reach the male rove beetles. Additionally, male and female rove beetles were characterized by distinctive microbiota in different organs, likely reflecting different functions and/or adaptation processes. Evidence of the extension of P. fuscipes microbiome from the environmental paradigm to the pathobiome was also presented herein. A comprehensive survey of P. fuscipes microbiome components may eventually lead to ecological insights into the production and utilization of defensive compound of pederin and also the management of linear dermatitis with the use of available antibiotics against bacterial pathogens released by the beetles. Video Abstract.}, } @article {pmid36669676, year = {2023}, author = {Awad, M and Piálková, R and Haelewaters, D and Nedvěd, O}, title = {Infection patterns of Harmonia axyridis (Coleoptera: Coccinellidae) by ectoparasitic microfungi and endosymbiotic bacteria.}, journal = {Journal of invertebrate pathology}, volume = {197}, number = {}, pages = {107887}, doi = {10.1016/j.jip.2023.107887}, pmid = {36669676}, issn = {1096-0805}, mesh = {Female ; Animals ; *Coleoptera/microbiology ; *Ascomycota ; Fertility ; *Spiroplasma ; Seasons ; *Wolbachia ; }, abstract = {The invasive alien ladybird Harmonia axyridis (Coleoptera: Coccinellidae) hosts a wide range of natural enemies. Many observations have been done in nature but experimental studies of interactions of multiple enemies on Ha. axyridis are rare. In light of this knowledge gap, we tested whether the host phenotype and presence of bacterial endosymbionts Spiroplasma and Wolbachia affected parasitism of Ha. axyridis by the ectoparasitic fungus Hesperomyces harmoniae (Ascomycota: Laboulbeniales). We collected 379 Ha. axyridis in the Czech Republic, processed specimens, including screening for He. harmoniae and a molecular assessment for bacteria, and calculated fecundity and hatchability of females. We found that high hatchability rate (71 %) was conditioned by high fecundity (20 eggs daily or more). The average parasite prevalence of He. harmoniae was 53 %, while the infection rate of Spiroplasma was 73 % in ladybirds that survived in winter conditions. Wolbachia was only present in 2 % of the analyzed ladybirds. Infection by either He. harmoniae or Spiroplasma did not differ among host color morphs. In the novemdecimsignata morph, younger individuals (with orange elytra) were more heavily parasitized compared to old ones (with red elytra). Fecundity and hatchability rate of females were unaffected by infection with either He. harmoniae or Spiroplasma. However, female ladybirds co-infected with He. harmoniae and Spiroplasma had a significantly lower fecundity and hatchability compared to females with only one or no symbiont.}, } @article {pmid36661997, year = {2023}, author = {Sevestre, J and Lemrabott, MAO and Bérenger, JM and Zan Diarra, A and Ould Mohamed Salem Boukhary, A and Parola, P}, title = {Detection of Arthropod-Borne Bacteria and Assessment of MALDI-TOF MS for the Identification of Field-Collected Immature Bed Bugs from Mauritania.}, journal = {Insects}, volume = {14}, number = {1}, pages = {}, pmid = {36661997}, issn = {2075-4450}, support = {PDH-2-NRBC-2-B-2113//Agence Nationale de la Recherche/ ; ANR-11-IDEX-0001-02//Agence Nationale de la Recherche/ ; }, abstract = {Human infestations by bed bugs have upsurged globally in recent decades, including in African countries, where recent reports pointed out an increase in infestation. Sympatric dwelling has been described for two species of bed bug parasitizing humans: Cimex hemipterus (the tropical bed bug) and C. lectularius. Identification of these two species is based on morphological characteristics, and gene sequencing, and may also rely on Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS). The present work aimed to assess whether MALDI-TOF MS was applicable for species level identification of immature stages of Cimex. Arthropods were collected in domestic settings in Nouakchott, Mauritania. Identification used morphological keys and MALDI-TOF MS identification was assessed for immature stages. Quantitative PCR and sequencing assays were used to detect arthropod-associated bacteria in each specimen. A total of 92 arthropods were collected, all morphologically identified as C. hemipterus (32 males, 14 females and 45 immature stages). A total of 35/45 specimens produced good quality MALDI-TOF MS spectra. Analysis allowed species level identification of all immature C. hemipterus after their spectra were entered into our in-house MALDI-TOF MS arthropod spectra database. Molecular screening allowed detection of Wolbachia DNA in each specimen. These results suggested that MALDI-TOF MS is a reliable tool for species level identification of Cimex specimens, including immature specimens. Future studies should assess this approach on larger panels of immature specimens for different Cimex species and focus on the precise staging of their different immature developmental stages.}, } @article {pmid36650806, year = {2023}, author = {Vicencio, D and Vasilieva, O and Gajardo, P}, title = {Monotonicity properties arising in a simple model of Wolbachia invasion for wild mosquito populations.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {20}, number = {1}, pages = {1148-1175}, doi = {10.3934/mbe.2023053}, pmid = {36650806}, issn = {1551-0018}, mesh = {Animals ; Humans ; *Wolbachia ; *Aedes ; Reproduction ; Fertility ; Population Dynamics ; }, abstract = {In this paper, we propose a simplified bidimensional Wolbachia infestation model in a population of Aedes aegypti mosquitoes, preserving the main features associated with the biology of this species that can be found in higher-dimensional models. Namely, our model represents the maternal transmission of the Wolbachia symbiont, expresses the reproductive phenotype of cytoplasmic incompatibility, accounts for different fecundities and mortalities of infected and wild insects, and exhibits the bistable nature leading to the so-called principle of competitive exclusion. Using tools borrowed from monotone dynamical system theory, in the proposed model, we prove the existence of an invariant threshold manifold that allows us to provide practical recommendations for performing single and periodic releases of Wolbachia-carrying mosquitoes, seeking the eventual elimination of wild insects that are capable of transmitting infections to humans. We illustrate these findings with numerical simulations using parameter values corresponding to the wMelPop strain of Wolbachia that is considered the best virus blocker but induces fitness loss in its carriers. In these tests, we considered multiple scenarios contrasting a periodic release strategy against a strategy with a single inundative release, comparing their effectiveness. Our study is presented as an expository and mathematically accessible tool to study the use of Wolbachia-based biocontrol versus more complex models.}, } @article {pmid36650789, year = {2023}, author = {Li, Y and Zhao, H and Wang, K}, title = {Dynamics of an impulsive reaction-diffusion mosquitoes model with multiple control measures.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {20}, number = {1}, pages = {775-806}, doi = {10.3934/mbe.2023036}, pmid = {36650789}, issn = {1551-0018}, mesh = {Animals ; *Insecticides ; *Aedes ; Mosquito Control ; Mosquito Vectors ; *Wolbachia ; }, abstract = {It is well-known that mosquito control is one of the effective methods to reduce and prevent the transmission of mosquito-borne diseases. In this paper, we formulate a reaction-diffusion impulsive hybrid model incorporating Wolbachia, impulsively spraying of insecticides, spatial heterogeneity, and seasonality to investigate the control of mosquito population. The sufficient conditions for mosquito extinction or successful Wolbachia persistence in a population of natural mosquitoes are derived. More importantly, we give the estimations of the spraying times of insecticides during a period for achieving the mosquito extinction and population replacement in a special case. A global attractivity of the positive periodic solution is analyzed under appropriate conditions. Numerical simulations disclose that spatial heterogeneity and seasonality have significant impacts on the design of mosquitoes control strategies. It is suggested to combine biological control and chemical pulse control under certain situations to reduce the natural mosquitoes. Further, our results reveal that the establishment of a higher level of population replacement depends on the strain type of the Wolbachia and the high initial occupancy of the Wolbachia-infected mosquitoes.}, } @article {pmid36650067, year = {2023}, author = {Miao, YH and Huang, DW and Xiao, JH}, title = {Complicated expansion trajectories of insertion sequences and potential association with horizontal transfer of Wolbachia DNA.}, journal = {Zoological research}, volume = {44}, number = {2}, pages = {273-275}, pmid = {36650067}, issn = {2095-8137}, mesh = {Animals ; *DNA Transposable Elements ; *Wolbachia/genetics ; Phylogeny ; }, } @article {pmid36648231, year = {2023}, author = {Nevalainen, LB and Layton, EM and Newton, ILG}, title = {Wolbachia Promotes Its Own Uptake by Host Cells.}, journal = {Infection and immunity}, volume = {91}, number = {2}, pages = {e0055722}, pmid = {36648231}, issn = {1098-5522}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Wolbachia ; Vacuoles ; Actins ; Symbiosis ; Drosophila melanogaster/microbiology ; }, abstract = {Wolbachia pipientis is an incredibly widespread bacterial symbiont of insects, present in an estimated 25 to 52% of species worldwide. Wolbachia is faithfully maternally transmitted both in a laboratory setting and in the wild. In an established infection, Wolbachia is primarily intracellular, residing within host-derived vacuoles that are associated with the endoplasmic reticulum. However, Wolbachia also frequently transfers between host species, requiring an extracellular stage to its life cycle. Indeed, Wolbachia has been moved between insect species for the precise goal of controlling populations. The use of Wolbachia in this application requires that we better understand how it initiates and establishes new infections. Here, we designed a novel method for live tracking Wolbachia cells during infection using a combination of stains and microscopy. We show that live Wolbachia cells are taken up by host cells at a much faster rate than dead Wolbachia cells, indicating that Wolbachia bacteria play a role in their own uptake and that Wolbachia colonization is not just a passive process. We also show that the host actin cytoskeleton must be intact for this to occur and that drugs that disrupt the actin cytoskeleton effectively abrogate Wolbachia uptake. The development of this live infection assay will assist in future efforts to characterize Wolbachia factors used during host infection.}, } @article {pmid36642905, year = {2022}, author = {Zhou, XQ and Ma, J and Wang, RY and Wang, RH and Wu, YQ and Yang, XY and Chen, YJ and Tang, XN and Sun, ET}, title = {[Bacterial community diversity in Dermatophagoides farinae using high-throughput sequencing].}, journal = {Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control}, volume = {34}, number = {6}, pages = {630-634}, doi = {10.16250/j.32.1374.2022105}, pmid = {36642905}, issn = {1005-6661}, support = {31870352//National Natural Science Foundation of China/ ; S202110368010//Anhui Provincial University Students' Innovation and Entrepreneurship Training Program/ ; }, mesh = {Humans ; Animals ; *Dermatophagoides farinae/genetics ; RNA, Ribosomal, 16S/genetics ; Bacteria/genetics ; High-Throughput Nucleotide Sequencing ; *Microbiota ; Phylogeny ; }, abstract = {OBJECTIVE: To investigate the bacterial community diversity in Dermatophagoides farinae.

METHODS: Laboratory-cultured D. farinae was collected, and the composition of microbial communities was determined by sequence analyses of the V4 region in the bacterial 16S ribosomal RNA (16S rRNA) gene on an Illumina PE250 high-throughput sequencing platform. Following quality control and filtering of the raw sequence files, valid reads were obtained and subjected to operational taxonomic units (OTU) clustering and analysis of the composition of microbial communities and alpha diversity index using the Usearch software, Silva database, and Mothur software.

RESULTS: A total of 187 616 valid reads were obtained, and 469 OTUs were clustered based on a sequence similarity of more than 97%. OTU annotation showed that the bacteria in D. farinae belonged to 26 phyla, 43 classes, 100 orders, 167 families and 284 genera. The bacteria in D. farinae were mainly annotated to five phyla of Proteobacteria, Firmicutes, Bacteroidota, Actinobacteriota, and Acidobacteriota, with Proteobacteria as the dominant phylum, and mainly annotated to five dominant genera of Ralstonia, norank-f-Mitochondria, Staphylococcus and Sphingomonas, with Wolbachia identified in the non-dominant genus.

CONCLUSIONS: A high diversity is identified in the composition of the bacterial community in D. farinae, and there are differences in bacterial community diversity and abundance among D. farinae.}, } @article {pmid36636344, year = {2023}, author = {Hussain, M and Zhang, G and Leitner, M and Hedges, LM and Asgari, S}, title = {Wolbachia RNase HI contributes to virus blocking in the mosquito Aedes aegypti.}, journal = {iScience}, volume = {26}, number = {1}, pages = {105836}, pmid = {36636344}, issn = {2589-0042}, abstract = {The endosymbiotic bacterium Wolbachia pipientis blocks replication of several arboviruses in transinfected Aedes aegypti mosquitoes. However, the mechanism of virus blocking remains poorly understood. Here, we characterized an RNase HI gene from Wolbachia, which is rapidly induced in response to dengue virus (DENV) infection. Knocking down w RNase HI using antisense RNA in Wolbachia-transinfected mosquito cell lines and A. aegypti mosquitoes led to increased DENV replication. Furthermore, overexpression of wRNase HI, in the absence of Wolbachia, led to reduced replication of a positive sense RNA virus, but had no effect on a negative sense RNA virus, a familiar scenario in Wolbachia-infected cells. Altogether, our results provide compelling evidence for the missing link between early Wolbachia-mediated virus blocking and degradation of viral RNA. These findings and the successful pioneered knockdown of Wolbachia genes using antisense RNA in cell line and mosquitoes enable new ways to manipulate and study the complex endosymbiont-host interactions.}, } @article {pmid36629101, year = {2023}, author = {Dou, W and Sun, B and Miao, Y and Huang, D and Xiao, J}, title = {Single-cell transcriptome sequencing reveals Wolbachia-mediated modification in early stages of Drosophila spermatogenesis.}, journal = {Proceedings. Biological sciences}, volume = {290}, number = {1990}, pages = {20221963}, pmid = {36629101}, issn = {1471-2954}, mesh = {Animals ; Male ; *Drosophila/genetics ; *Wolbachia/genetics ; Drosophila melanogaster/genetics/microbiology ; Transcriptome ; Semen ; Spermatogenesis ; Cytoplasm/microbiology ; }, abstract = {Wolbachia are the most widely distributed intracellular bacteria, and their most common effect on host phenotype is cytoplasmic incompatibility (CI). A variety of models have been proposed to decipher the molecular mechanism of CI, among which the host modification (HM) model predicts that Wolbachia effectors play an important role in sperm modification. However, owing to the complexity of spermatogenesis and testicular cell-type heterogeneity, whether Wolbachia have different effects on cells at different stages of spermatogenesis or whether these effects are linked with CI remains unknown. Therefore, we used single-cell RNA sequencing to analyse gene expression profiles in adult male Drosophila testes that were infected or uninfected by Wolbachia. We found that Wolbachia significantly affected the proportion of different types of germ cells and affected multiple metabolic pathways in germ cells. Most importantly, Wolbachia had the greatest impact on germline stem cells, resulting in dysregulated expression of genes related to DNA compaction, and Wolbachia infection also influenced the histone-to-protamine transition in the late stage of sperm development. These results support the HM model and suggest that future studies on Wolbachia-induced CI should focus on cells in the early stages of spermatogenesis.}, } @article {pmid36628964, year = {2023}, author = {Durand, S and Lheraud, B and Giraud, I and Bech, N and Grandjean, F and Rigaud, T and Peccoud, J and Cordaux, R}, title = {Heterogeneous distribution of sex ratio distorters in natural populations of the isopod Armadillidium vulgare.}, journal = {Biology letters}, volume = {19}, number = {1}, pages = {20220457}, pmid = {36628964}, issn = {1744-957X}, mesh = {Male ; Animals ; Female ; *Isopoda/genetics ; Sex Ratio ; Haplotypes ; Europe ; Japan ; *Wolbachia/genetics ; }, abstract = {In the isopod Armadillidium vulgare, many females produce progenies with female-biased sex ratios, owing to two feminizing sex ratio distorters (SRD): Wolbachia endosymbionts and the f element. We investigated the distribution and population dynamics of these SRD and mitochondrial DNA variation in 16 populations from Europe and Japan. Confirming and extending results from the 1990s, we found that the SRD are present at variable frequencies in populations and that the f element is overall more frequent than Wolbachia. The two SRD never co-occur at high frequency in any population, suggesting an apparent mutual exclusion. We also detected Wolbachia or the f element in some males, which probably reflects insufficient titer to induce feminization or presence of masculinizing alleles. Our results are consistent with a single integration event of a Wolbachia genome in the A. vulgare genome at the origin of the f element, which contradicts an earlier hypothesis of frequent losses and gains. We identified strong linkage between Wolbachia strains and mitochondrial haplotypes, but no association between the f element and mitochondrial background. Our results open new perspectives on SRD evolutionary dynamics in A. vulgare, the evolution of genetic conflicts and their impact on the variability of sex determination systems.}, } @article {pmid36589876, year = {2022}, author = {Uni, S and Mat Udin, AS and Tan, PE and Rodrigues, J and Martin, C and Junker, K and Agatsuma, T and Low, VL and Lim, YA and Saijuntha, W and Omar, H and Zainuri, NA and Fukuda, M and Kimura, D and Matsubayashi, M and Uga, S and Takaoka, H and Azirun, MS and Ramli, R}, title = {Description and molecular characterisation of Pelecitus copsychi Uni, Mat Udin & Martin n. sp. (Nematoda: Onchocercidae) from the white-rumped shama Copsychus malabaricus (Scopoli) (Passeriformes: Muscicapidae) of Pahang, Malaysia.}, journal = {Current research in parasitology & vector-borne diseases}, volume = {2}, number = {}, pages = {100078}, pmid = {36589876}, issn = {2667-114X}, abstract = {Species of the genus Pelecitus Railliet & Henry, 1910 the most widely distributed avian filariae in Africa and South America. Zoonotic cases in humans were reported in South America. While investigating the filarial fauna of wild animals in Malaysia, we discovered an undescribed filaria from the swollen footpad of the left leg of Copsychus malabaricus (Scopoli) in Pahang, Peninsular Malaysia. Adults of both sexes have a corkscrew-shaped body. Based on comparison of their morphological characteristics (i.e. pre-oesophageal cuticular ring distinct, oesophagus divided, vulva protuberant and situated at the level of anterior half of oesophagus, spicules strongly sclerotized and left spicule with broad blade) with other Pelecitus species, they are here described as Pelecitus copsychi Uni, Mat Udin & Martin n. sp. Multi-locus sequence analyses based on seven genes (12S rDNA, cox1, 18S rDNA, 28S rDNA, MyoHC, rbp1 and hsp70) were performed to determine the phylogenetic position of the new species. The calculated p-distance between the cox1 gene sequences for P. copsychi n. sp. and Pelecitus fulicaeatrae (Diesing, 1861) was 14.1%. Intraspecific genetic variation between two individuals of the new species was 0.4%. In both the Bayesian inference and maximum-likelihood trees, P. copsychi n. sp. was positioned in the second clade of ONC5, containing three genera of the subfamily Dirofilariinae (Foleyella Seurat, 1917, Pelecitus and Loa Stiles, 1905). Immunostaining and molecular analyses remained negative for the presence of Wolbachia endosymbionts. Our findings corroborate the division of the subfamily Dirofilariinae into ONC3 with Dirofilaria Railliet & Henry, 1911 and ONC5 with Pelecitus.}, } @article {pmid36580820, year = {2023}, author = {Martinez-Cruz, C and Arenas-Monreal, L and Gomez-Dantes, H and Villegas-Chim, J and Barrera-Fuentes Gloria, A and Toledo-Romani Maria, E and Pavia-Ruz, N and Che-Mendoza, A and Manrique-Saide, P}, title = {Educational intervention for the control of Aedes aegypti with Wolbachia in Yucatan, Mexico.}, journal = {Evaluation and program planning}, volume = {97}, number = {}, pages = {102205}, doi = {10.1016/j.evalprogplan.2022.102205}, pmid = {36580820}, issn = {1873-7870}, mesh = {Animals ; Humans ; Male ; Child ; *Aedes/microbiology ; *Wolbachia ; Mexico ; Mosquito Vectors/microbiology ; Program Evaluation ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {UNLABELLED: The implementation of new control strategies for Aedes aegypti (Ae. Aegpyti), a vector of dengue, chikungunya, and Zika viruses, requires communities to adopt specific behaviors to achieve the success of these innovations.

AIM: We evaluated the effect of an educational intervention based on the Precede-Proceed Model (PPM) and the Diffusion of Innovations Theory (DIT) for the control and prevention of diseases transmitted by Ae. aegypti through release of male mosquitoes infected with Wolbachia bacteria in a suburban town in Yucatan, Mexico.

MATERIAL AND METHODS: From July 2019 to February 2020, a quasi-experimental study was carried out through an educational intervention (pre- and post-measurements) using quantitative-qualitative techniques, in a Yucatan suburban town where male mosquitoes with Wolbachia were released for the suppression of Ae. aegypti populations. Eleven educational workshops were attended by heads of household (n = 19) and schoolchildren (n = 11). Other 136 heads of household not attending the workshops received information individually.

RESULTS: The educational intervention had a significant effect on the mean scores of the contributing and behavioral factors for adoption of innovation (p < 0.05) in the pre- and post-intervention measurements.

CONCLUSION: Innovative methods for the control and prevention of diseases related to Aedes aegypti can be strengthened through educational interventions supported by sound methodologies.

DESCRIPTORS: Community health education, Aedes aegypti, Wolbachia, Mexico.}, } @article {pmid36575628, year = {2023}, author = {Kil, EJ and Kim, D}, title = {The small brown planthopper (Laodelphax striatellus) as a vector of the rice stripe virus.}, journal = {Archives of insect biochemistry and physiology}, volume = {112}, number = {2}, pages = {e21992}, doi = {10.1002/arch.21992}, pmid = {36575628}, issn = {1520-6327}, support = {PJ01556601//Rural Development Administration/ ; }, mesh = {Animals ; *Tenuivirus/genetics ; Insect Vectors/genetics ; *Hemiptera/genetics ; Insecta/genetics ; Gene Expression Profiling ; Viral Proteins/metabolism ; *Oryza ; }, abstract = {The small brown planthopper, Laodelphax striatellus, is a destructive pest insect found in rice fields. L. striatellus not only directly feeds on the phloem sap of rice but also transmits various viruses, such as rice stripe virus (RSV) and rice black-streaked dwarf virus, resulting in serious loss of rice production. RSV is a rice-infecting virus that is found mainly in Korea, China, and Japan. To develop novel strategies to control L. striatellus and L. striatellus-transmitted viruses, various studies have been conducted, based on vector biology, interactions between vectors and pathogens, and omics, including transcriptomics, proteomics, and metabolomics. In this review, we discuss the roles of saliva proteins during phloem sap-sucking and virus transmission, the diversity and role of the microbial community in L. striatellus, the profile and molecular mechanisms of insecticide resistance, classification of L. striatellus-transmitted RSV, its host range and symptoms, its genome composition and roles of virus-derived proteins, its distribution, interactions with L. striatellus, and resistance and control, to suggest future directions for integrated pest management to control L. striatellus and L. striatellus-transmitted viruses.}, } @article {pmid36575240, year = {2022}, author = {Zeng, Q and She, L and Yuan, H and Luo, Y and Wang, R and Mao, W and Wang, W and She, Y and Wang, C and Shi, M and Cao, T and Gan, R and Li, Y and Zhou, J and Qian, W and Hu, S and Wang, Y and Zheng, X and Li, K and Bai, L and Pan, X and Xi, Z}, title = {A standalone incompatible insect technique enables mosquito suppression in the urban subtropics.}, journal = {Communications biology}, volume = {5}, number = {1}, pages = {1419}, pmid = {36575240}, issn = {2399-3642}, mesh = {Animals ; Male ; Female ; Mosquito Control/methods ; *Aedes ; Population Dynamics ; Larva ; *Dengue/prevention & control ; }, abstract = {The strong suppression of Aedes albopictus on two Guangzhou islands in China has been successfully achieved by releasing males with an artificial triple-Wolbachia infection. However, it requires the use of radiation to sterilize residual females to prevent population replacement. To develop a highly effective tool for dengue control, we tested a standalone incompatible insect technique (IIT) to control A. albopictus in the urban area of Changsha, an inland city where dengue recently emerged. Male mosquitoes were produced in a mass rearing facility in Guangzhou and transported over 670 km under low temperature to the release site. After a once-per-week release with high numbers of males (phase I) and a subsequent twice-per-week release with low numbers of males (phase II), the average numbers of hatched eggs and female adults collected weekly per trap were reduced by 97% and 85%, respectively. The population suppression caused a 94% decrease in mosquito biting at the release site compared to the control site. Remarkably, this strong suppression was achieved using only 28% of the number of males released in a previous trial. Despite the lack of irradiation to sterilize residual females, no triple-infected mosquitoes were detected in the field post release based on the monitoring of adult and larval A. albopictus populations for two years, indicating that population replacement was prevented. Our results support the feasibility of implementing a standalone IIT for dengue control in urban areas.}, } @article {pmid36555851, year = {2022}, author = {Korenskaia, AE and Shishkina, OD and Klimenko, AI and Andreenkova, OV and Bobrovskikh, MA and Shatskaya, NV and Vasiliev, GV and Gruntenko, NE}, title = {New Wolbachia pipientis Genotype Increasing Heat Stress Resistance of Drosophila melanogaster Host Is Characterized by a Large Chromosomal Inversion.}, journal = {International journal of molecular sciences}, volume = {23}, number = {24}, pages = {}, pmid = {36555851}, issn = {1422-0067}, support = {21-14-00090//Russian Science Foundation/ ; FWNR-2022-0019//MINISTRY OF SCIENCE AND HIGHER EDUCATION OF THE RUSSIAN FEDERATION/ ; }, mesh = {Animals ; *Drosophila melanogaster/genetics/microbiology ; *Wolbachia/genetics ; Chromosome Inversion ; Genotype ; Heat-Shock Response/genetics ; Symbiosis ; }, abstract = {The maternally transmitted endocellular bacteria Wolbachia is a well-known symbiont of insects, demonstrating both negative and positive effects on host fitness. The previously found Wolbachia strain wMelPlus is characterized by a positive effect on the stress-resistance of its host Drosophila melanogaster, under heat stress conditions. This investigation is dedicated to studying the genomic underpinnings of such an effect. We sequenced two closely related Wolbachia strains, wMelPlus and wMelCS112, assembled their complete genomes, and performed comparative genomic analysis engaging available Wolbachia genomes from the wMel and wMelCS groups. Despite the two strains under study sharing very close gene-composition, we discovered a large (>1/6 of total genome) chromosomal inversion in wMelPlus, spanning through the region that includes the area of the inversion earlier found in the wMel group of Wolbachia genotypes. A number of genes in unique inversion blocks of wMelPlus were identified that might be involved in the induction of a stress-resistant phenotype in the host. We hypothesize that such an inversion could rearrange established genetic regulatory-networks, causing the observed effects of such a complex fly phenotype as a modulation of heat stress resistance. Based on our findings, we propose that wMelPlus be distinguished as a separate genotype of the wMelCS group, named wMelCS3.}, } @article {pmid36554999, year = {2022}, author = {Kaszyca-Taszakowska, N and Depa, Ł}, title = {Microbiome of the Aphid Genus Dysaphis Börner (Hemiptera: Aphidinae) and Its Relation to Ant Attendance.}, journal = {Insects}, volume = {13}, number = {12}, pages = {}, pmid = {36554999}, issn = {2075-4450}, support = {no UMO-2018/29/N/NZ8/00360//National Science Centre, Poland/ ; }, abstract = {Among mutualistic relationships of aphids with other organisms, there are two that seem to be of major importance: trophobiosis with ants and endosymbiosis of bacteria. While the former is well studied, the latter is the subject of an increasing amount of research constantly revealing new aspects of this symbiosis. Here, we studied the possible influence of ant attendance on the composition of aphid microbiota on primary and secondary hosts exploited by the aphid genus Dysaphis. The microbiome of 44 samples representing 12 aphid species was studied using an Illumina HiSeq 4000 with the V3-V4 region of 16S rRNA. The results showed a higher abundance of common facultative symbionts (Serratia, Regiella, Fukatsuia) in aphid species unattended by ants, but also on secondary hosts. However, in colonies attended by ants, the general species composition of bacterial symbionts was more rich in genera than in unattended colonies (Wolbachia, Gilliamella, Spiroplasma, Sphingomonas, Pelomonas). The results indicate a huge variability of facultative symbionts without clear correlation with ant attendance or aphid species. The possibility of multiple routes of bacterial infection mediated by ant-made environmental conditions is discussed.}, } @article {pmid36530420, year = {2022}, author = {Zhang, H and Gao, J and Ma, Z and Liu, Y and Wang, G and Liu, Q and Du, Y and Xing, D and Li, C and Zhao, T and Jiang, Y and Dong, Y and Guo, X and Zhao, T}, title = {Wolbachia infection in field-collected Aedes aegypti in Yunnan Province, southwestern China.}, journal = {Frontiers in cellular and infection microbiology}, volume = {12}, number = {}, pages = {1082809}, pmid = {36530420}, issn = {2235-2988}, mesh = {Animals ; *Wolbachia/genetics ; *Aedes/microbiology ; Phylogeny ; China/epidemiology ; DNA Primers ; }, abstract = {BACKGROUND: Wolbachia is gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Recent reports reveal the natural infection of Wolbachia in Aedes Aegypti in Malaysia, India, Philippines, Thailand and the United States. At present, none of Wolbachia natural infection in Ae. aegypti has been reported in China.

METHODS: A total of 480 Ae. aegypti adult mosquitoes were collected from October and November 2018 based on the results of previous investigations and the distribution of Ae. aegypti in Yunnan. Each individual sample was processed and screened for the presence of Wolbachia by PCR with wsp primers. Phylogenetic trees for the wsp gene was constructed using the neighbour-joining method with 1,000 bootstrap replicates, and the p-distance distribution model of molecular evolution was applied.

RESULTS: 24 individual adult mosquito samples and 10 sample sites were positive for Wolbachia infection. The Wolbachia infection rate (IR) of each population ranged from 0 - 41.7%. The infection rate of group A alone was 0%-10%, the infection rate of group B alone was 0%-7.7%, and the infection rate of co-infection with A and B was 0-33.3%.

CONCLUSIONS: Wolbachia infection in wild Ae. aegypti in China is the first report based on PCR amplification of the Wolbachia wsp gene. The Wolbachia infection is 5%, and the wAlbA and wAlbB strains were found to be prevalent in the natural population of Ae. aegypti in Yunnan Province.}, } @article {pmid36528590, year = {2022}, author = {Ong, J and Ho, SH and Soh, SXH and Wong, Y and Ng, Y and Vasquez, K and Lai, YL and Setoh, YX and Chong, CS and Lee, V and Wong, JCC and Tan, CH and Sim, S and Ng, LC and Lim, JT}, title = {Assessing the efficacy of male Wolbachia-infected mosquito deployments to reduce dengue incidence in Singapore: study protocol for a cluster-randomized controlled trial.}, journal = {Trials}, volume = {23}, number = {1}, pages = {1023}, pmid = {36528590}, issn = {1745-6215}, mesh = {Animals ; Male ; Humans ; *Wolbachia ; Mosquito Control/methods ; *Dengue/epidemiology/prevention & control ; Mosquito Vectors ; Incidence ; Seroepidemiologic Studies ; Singapore/epidemiology ; *Aedes ; Randomized Controlled Trials as Topic ; }, abstract = {BACKGROUND: Dengue is a severe environmental public health challenge in tropical and subtropical regions. In Singapore, decreasing seroprevalence and herd immunity due to successful vector control has paradoxically led to increased transmission potential of the dengue virus. We have previously demonstrated that incompatible insect technique coupled with sterile insect technique (IIT-SIT), which involves the release of X-ray-irradiated male Wolbachia-infected mosquitoes, reduced the Aedes aegypti population by 98% and dengue incidence by 88%. This novel vector control tool is expected to be able to complement current vector control to mitigate the increasing threat of dengue on a larger scale. We propose a multi-site protocol to study the efficacy of IIT-SIT at reducing dengue incidence.

METHODS/DESIGN: The study is designed as a parallel, two-arm, non-blinded cluster-randomized (CR) controlled trial to be conducted in high-rise public housing estates in Singapore, an equatorial city-state. The aim is to determine whether large-scale deployment of male Wolbachia-infected Ae. aegypti mosquitoes can significantly reduce dengue incidence in intervention clusters. We will use the CR design, with the study area comprising 15 clusters with a total area of 10.9 km[2], covering approximately 722,204 residents in 1713 apartment blocks. Eight clusters will be randomly selected to receive the intervention, while the other seven will serve as non-intervention clusters. Intervention efficacy will be estimated through two primary endpoints: (1) odds ratio of Wolbachia exposure distribution (i.e., probability of living in an intervention cluster) among laboratory-confirmed reported dengue cases compared to test-negative controls and (2) laboratory-confirmed reported dengue counts normalized by population size in intervention versus non-intervention clusters.

DISCUSSION: This study will provide evidence from a multi-site, randomized controlled trial for the efficacy of IIT-SIT in reducing dengue incidence. The trial will provide valuable information to estimate intervention efficacy for this novel vector control approach and guide plans for integration into national vector control programs in dengue-endemic settings.

TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT05505682 . Registered on 16 August 2022. Retrospectively registered.}, } @article {pmid36524932, year = {2023}, author = {Schmidt, TL and Elfekih, S and Cao, LJ and Wei, SJ and Al-Fageeh, MB and Nassar, M and Al-Malik, A and Hoffmann, AA}, title = {Close Kin Dyads Indicate Intergenerational Dispersal and Barriers.}, journal = {The American naturalist}, volume = {201}, number = {1}, pages = {65-77}, doi = {10.1086/722175}, pmid = {36524932}, issn = {1537-5323}, mesh = {Humans ; Animals ; Female ; *Aedes/genetics ; Population Density ; *Wolbachia ; }, abstract = {AbstractThe movement of individuals through continuous space is typically constrained by dispersal ability and dispersal barriers. A range of approaches have been developed to investigate these. Kindisperse is a new approach that infers recent intergenerational dispersal (σ) from close kin dyads and appears particularly useful for investigating taxa that are difficult to observe individually. This study, focusing on the mosquito Aedes aegypti, shows how the same close kin data can also be used for barrier detection. We empirically demonstrate this new extension of the method using genome-wide sequence data from 266 Ae. aegypti. First, we use the spatial distribution of full-sib dyads collected within one generation to infer past movements of ovipositing female mosquitoes. These dyads indicated the relative barrier strengths of two roads and performed favorably against alternative genetic methods for detecting barriers. We then use Kindisperse to quantify recent intergenerational dispersal (σ=81.5-197.1 m generation[-1/2]) from the difference in variance between the sib and the first cousin spatial distributions and, from this, estimate effective population density (ρ=833-4,864 km[-2]). Dispersal estimates showed general agreement with those from mark-release-recapture studies. Barriers, σ, ρ, and neighborhood size (331-526) can inform forthcoming releases of dengue-suppressing Wolbachia bacteria into this mosquito population.}, } @article {pmid36502993, year = {2023}, author = {Venkataravanappa, V and Kodandaram, MH and Prasanna, HC and Reddy, MK and Reddy, CNL}, title = {Unraveling different begomoviruses, DNA satellites and cryptic species of Bemisia tabaci and their endosymbionts in vegetable ecosystem.}, journal = {Microbial pathogenesis}, volume = {174}, number = {}, pages = {105892}, doi = {10.1016/j.micpath.2022.105892}, pmid = {36502993}, issn = {1096-1208}, mesh = {Animals ; Vegetables ; Ecosystem ; *Begomovirus/genetics ; Crops, Agricultural/genetics ; *Hemiptera ; DNA ; Plant Diseases ; }, abstract = {Bemisia tabaci species complex contains more than 46 cryptic species. It has emerged as an important pest causing significant yield loss in many cultivated crops. This pest is also a vector for more than 100 species of begomoviruses, that are a major threat for the cultivation of many crops in different regions of the world. The relation between cryptic species of the B. tabaci species complex and associated begomoviruses that infect different crops remains unclear. In the present study, four cryptic species (Asia I, China 3, Asia II 5 and Asia II-1) of B. tabaci and four associated endosymbionts (Arsenophonus, Cardinium, Rickettsia and Wolbachia) were identified in different vegetable crops. The vector-based PCR detection revealed five different begomoviruses such as okra enation leaf curl virus (OELCuV), tomato leaf curl Palampur virus (ToLCPalV), squash leaf curl China virus (SLCCNV), chilli leaf curl virus (ChiLCuV), and tomato leaf curl New Delhi virus (ToLCNDV). Of these begomoviruses, the maximum infection rate was observed (9.1%) for OELCuV, followed by 7.3% for ToLCNDV. The infection rate of the other three viruses (SLCCNV, ChiLCuV, ToLCPalV) ranged from 0.9 to 2.7% in cryptic species of B. tabaci. Further, each cryptic species was infected with multiple virus species and the virus infection rate of Asia I, Asia II-5, China 3 and Asia II-1 was 21.2%, 15.1%, 15.1% and 0.6% respectively. Similarly, in case of betasatellites the highest infection rate was 12% for ToLCBDB, followed by 6% for OLCuB and PaLCB. With regard to alphasatellites, the highest infection rate was 18.2% for AEV and 3% for CLCuMuA. This study demonstrates the distribution of cryptic species of whitefly and their endosymbionts, and associated begomoviruses and DNA satellites in vegetable ecosystem. We believe that the information generated here is useful for evolving an effective pest management strategies for vegetable production.}, } @article {pmid36499640, year = {2022}, author = {Andreenkova, OV and Shishkina, OD and Klimenko, AI and Korenskaia, AE and Bobrovskikh, MA and Shatskaya, NV and Vasiliev, GV and Gruntenko, NE}, title = {Easy and Effective Method for Extracting and Purifying Wolbachia Genomic DNA.}, journal = {International journal of molecular sciences}, volume = {23}, number = {23}, pages = {}, pmid = {36499640}, issn = {1422-0067}, support = {21-14-00090//Russian Science Foundation/ ; FWNR-2022-0019//MINISTRY OF SCIENCE AND HIGHER EDUCATION OF THE RUSSIAN FEDERATION/ ; }, mesh = {Animals ; *Wolbachia/genetics ; Drosophila melanogaster/genetics/microbiology ; Sequence Analysis, DNA ; Chromosome Mapping ; DNA ; Symbiosis ; }, abstract = {A number of methods for extracting the DNA of maternally inherited obligate intracellular bacteria Wolbachia from an insect host and its subsequent purification have been described in previous scholarship. As Wolbachia is present in the hosts' organisms in rather low quantities, these techniques used to be quite labor-intensive. For this paper, we analyzed them in detail, searched for a possibility to simplify and accelerate the protocol, and proposed an easy and effective method for isolating Wolbachia DNA from Drosophila melanogaster with a purity sufficient for genomic sequencing. Our method involves the centrifugation of homogenized flies or just their ovaries, as the most Wolbachia-enriched tissue, followed by the filtration of homogenate and extraction of DNA using a modified version of the Livak buffer protocol. The proportion of Wolbachia DNA in the total DNA was quantified based on the results of sequencing with the use of the Illumina MiSeq platform and a pipeline of bioinformatic analysis. For the two analyzed D. melanogaster lines infected with two different Wolbachia strains, the proportion was at least 68 and 94%, respectively.}, } @article {pmid36496327, year = {2023}, author = {Zhu, Z and Liu, Y and Hu, H and Wang, GH}, title = {Nasonia-microbiome associations: a model for evolutionary hologenomics research.}, journal = {Trends in parasitology}, volume = {39}, number = {2}, pages = {101-112}, doi = {10.1016/j.pt.2022.11.005}, pmid = {36496327}, issn = {1471-5007}, mesh = {Animals ; Biological Evolution ; *Wasps/genetics ; Insecta/genetics ; *Microbiota/genetics ; Symbiosis ; }, abstract = {In recent years, with the development of microbial research technologies, microbiota research has received widespread attention. The parasitoid wasp genus Nasonia is a good model organism for studying insect behavior, development, evolutionary genetics, speciation, and symbiosis. This review describes key advances and progress in the field of the Nasonia-microbiome interactions. We provide an overview of the advantages of Nasonia as a model organism for microbiome studies, list research methods to study the Nasonia microbiome, and discuss recent discoveries in Nasonia microbiome research. This summary of the complexities of Nasonia-microbiome relationships will help to contribute to a better understanding of the interactions between animals and their microbiomes and establish a clear research direction for Nasonia-microbiome interactions in the future.}, } @article {pmid36476840, year = {2022}, author = {Nakabachi, A and Inoue, H and Hirose, Y}, title = {High-resolution Microbiome Analyses of Nine Psyllid Species of the Family Triozidae Identified Previously Unrecognized but Major Bacterial Populations, including Liberibacter and Wolbachia of Supergroup O.}, journal = {Microbes and environments}, volume = {37}, number = {4}, pages = {}, pmid = {36476840}, issn = {1347-4405}, mesh = {Humans ; Animals ; Liberibacter ; *Wolbachia/genetics ; *Hemiptera ; RNA, Ribosomal, 16S/genetics ; Europe ; }, abstract = {Psyllids (Hemiptera: Sternorrhyncha: Psylloidea) are plant sap-sucking insects that include important agricultural pests. To obtain insights into the ecological and evolutionary behaviors of microbes, including plant pathogens, in Psylloidea, high-resolution ana-lyses of the microbiomes of nine psyllid species belonging to the family Triozidae were performed using high-throughput amplicon sequencing of the 16S rRNA gene. Analyses identified various bacterial populations, showing that all nine psyllids have at least one secondary symbiont, along with the primary symbiont "Candidatus Carsonella ruddii" (Gammaproteobacteria: Oceanospirillales: Halomonadaceae). The majority of the secondary symbionts were gammaproteobacteria, particularly those of the order Enterobacterales, which included Arsenophonus and Serratia symbiotica, a bacterium formerly recognized only as a secondary symbiont of aphids (Hemiptera: Sternorrhyncha: Aphidoidea). The non-Enterobacterales gammaproteobacteria identified in the present study were Diplorickettsia (Diplorickettsiales: Diplorickettsiaceae), a potential human pathogen, and Carnimonas (Oceanospirillales: Halomonadaceae), a lineage detected for the first time in Psylloidea. Regarding alphaproteobacteria, the potential plant pathogen "Ca. Liberibacter europaeus" (Rhizobiales: Rhizobiaceae) was detected for the first time in Epitrioza yasumatsui, which feeds on the Japanese silverberry Elaeagnus umbellata (Elaeagnaceae), an aggressive invasive plant in the United States and Europe. Besides the detection of Wolbachia (Rickettsiales: Anaplasmataceae) of supergroup B in three psyllid species, a lineage belonging to supergroup O was identified for the first time in Psylloidea. These results suggest the rampant transfer of bacterial symbionts among animals and plants, thereby providing deeper insights into the evolution of interkingdom interactions among multicellular organisms and bacteria, which will facilitate the control of pest psyllids.}, } @article {pmid36467729, year = {2022}, author = {Fordjour, FA and Kwarteng, A}, title = {The filarial and the antibiotics: Single or combination therapy using antibiotics for filariasis.}, journal = {Frontiers in cellular and infection microbiology}, volume = {12}, number = {}, pages = {1044412}, pmid = {36467729}, issn = {2235-2988}, mesh = {Humans ; Anti-Bacterial Agents/therapeutic use ; *Filariasis/drug therapy ; *Nematode Infections ; Combined Modality Therapy ; *Wolbachia ; }, abstract = {Filarial infections caused by nematodes are one of the major neglected tropical diseases with public health concern. Although there is significant decrease in microfilariae (mf) prevalence following mass drug administration (IVM/DEC/ALB administration), this is transient, in that there is reported microfilaria repopulation 6-12 months after treatment. Wolbachia bacteria have been recommended as a novel target presenting antibiotic-based treatment for filarial disease. Potency of antibiotics against filarial diseases is undoubtful, however, the duration for treatment remains a hurdle yet to be overcome in filarial disease treatment.}, } @article {pmid36467722, year = {2022}, author = {Hodosi, R and Kazimirova, M and Soltys, K}, title = {What do we know about the microbiome of I. ricinus?.}, journal = {Frontiers in cellular and infection microbiology}, volume = {12}, number = {}, pages = {990889}, pmid = {36467722}, issn = {2235-2988}, mesh = {Animals ; *Ixodes ; *Microbiota ; Coxiella ; Symbiosis ; *Rickettsia ; *Francisella tularensis ; }, abstract = {I. ricinus is an obligate hematophagous parasitic arthropod that is responsible for the transmission of a wide range of zoonotic pathogens including spirochetes of the genus Borrelia, Rickettsia spp., C. burnetii, Anaplasma phagocytophilum and Francisella tularensis, which are part the tick´s microbiome. Most of the studies focus on "pathogens" and only very few elucidate the role of "non-pathogenic" symbiotic microorganisms in I. ricinus. While most of the members of the microbiome are leading an intracellular lifestyle, they are able to complement tick´s nutrition and stress response having a great impact on tick´s survival and transmission of pathogens. The composition of the tick´s microbiome is not consistent and can be tied to the environment, tick species, developmental stage, or specific organ or tissue. Ovarian tissue harbors a stable microbiome consisting mainly but not exclusively of endosymbiotic bacteria, while the microbiome of the digestive system is rather unstable, and together with salivary glands, is mostly comprised of pathogens. The most prevalent endosymbionts found in ticks are Rickettsia spp., Ricketsiella spp., Coxiella-like and Francisella-like endosymbionts, Spiroplasma spp. and Candidatus Midichloria spp. Since microorganisms can modify ticks' behavior, such as mobility, feeding or saliva production, which results in increased survival rates, we aimed to elucidate the potential, tight relationship, and interaction between bacteria of the I. ricinus microbiome. Here we show that endosymbionts including Coxiella-like spp., can provide I. ricinus with different types of vitamin B (B2, B6, B7, B9) essential for eukaryotic organisms. Furthermore, we hypothesize that survival of Wolbachia spp., or the bacterial pathogen A. phagocytophilum can be supported by the tick itself since coinfection with symbiotic Spiroplasma ixodetis provides I. ricinus with complete metabolic pathway of folate biosynthesis necessary for DNA synthesis and cell division. Manipulation of tick´s endosymbiotic microbiome could present a perspective way of I. ricinus control and regulation of spread of emerging bacterial pathogens.}, } @article {pmid36466669, year = {2022}, author = {El Hamss, H and Maruthi, MN and Ally, HM and Omongo, CA and Wang, HL and van Brunschot, S and Colvin, J and Delatte, H}, title = {Spatio-temporal changes in endosymbiont diversity and composition in the African cassava whitefly, Bemisia tabaci SSA1.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {986226}, pmid = {36466669}, issn = {1664-302X}, abstract = {Sap-sucking insects, including whiteflies, are amongst the most devastating and widely distributed organisms on the planet. They are often highly invasive and endosymbiont communities within these insects help them adapt to new or changing environments. Bemisia tabaci (Gennadius; Hemiptera: Aleyrodidae) whitefly species are vectors of more than 500 known plant-viruses and harbour highly diverse endosymbionts communities. To date, however, whitefly-endosymbiont interactions, community structure and their spatio-temporal changes are still poorly understood. In this study, we investigated the spatio-temporal changes in the composition and diversity of bacterial endosymbionts in the agricultural crop pest whitefly species, Bemisia tabaci sub-Saharan Africa 1-subgroup 1 and 2 (SSA1-SG1 and SSA1-SG2). 16S rRNA amplicon sequencing analysis was carried out to characterise endosymbiont compositionsin field-collected SSA1 (SSA1-SG1 and SSA1-SG2) populations infesting cassava in Uganda in 1997 and 2017. We detected Portiera, Arsenophonus, Wolbachia, Hamiltonella and Hemipteriphilus, with Arsenophonus and Wolbachia infections being predominant. Hemipteriphilus and Hamiltonella frequencies were very low and were detected in seven and two samples, respectively. Bacterial diversity based on three independent parameters including Simpson index, number of haplotypes and Bray-Curtis dissimilarity matrix was significantly higher in 1997 than in 2017. This period also coincided with the advent of super-abundant cassava-whitefly populations on cassava crops in Uganda. We discuss how endosymbionts may influence the biology and behaviour of whiteflies leading to population explosions.}, } @article {pmid36460676, year = {2022}, author = {Ogunlade, ST and Adekunle, AI and McBryde, ES and Meehan, MT}, title = {Modelling the ecological dynamics of mosquito populations with multiple co-circulating Wolbachia strains.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {20826}, pmid = {36460676}, issn = {2045-2322}, mesh = {Animals ; *Wolbachia ; Mosquito Vectors ; *Aedes ; Body Temperature Regulation ; Climate Change ; }, abstract = {Wolbachia intracellular bacteria successfully reduce the transmissibility of arthropod-borne viruses (arboviruses) when introduced into virus-carrying vectors such as mosquitoes. Despite the progress made by introducing Wolbachia bacteria into the Aedes aegypti wild-type population to control arboviral infections, reports suggest that heat-induced loss-of-Wolbachia-infection as a result of climate change may reverse these gains. Novel, supplemental Wolbachia strains that are more resilient to increased temperatures may circumvent these concerns, and could potentially act synergistically with existing variants. In this article, we model the ecological dynamics among three distinct mosquito (sub)populations: a wild-type population free of any Wolbachia infection; an invading population infected with a particular Wolbachia strain; and a second invading population infected with a distinct Wolbachia strain from that of the first invader. We explore how the range of possible characteristics of each Wolbachia strain impacts mosquito prevalence. Further, we analyse the differential system governing the mosquito populations and the Wolbachia infection dynamics by computing the full set of basic and invasive reproduction numbers and use these to establish stability of identified equilibria. Our results show that releasing mosquitoes with two different strains of Wolbachia did not increase their prevalence, compared with a single-strain Wolbachia-infected mosquito introduction and only delayed Wolbachia dominance.}, } @article {pmid36450768, year = {2022}, author = {Lucati, F and Delacour, S and Palmer, JRB and Caner, J and Oltra, A and Paredes-Esquivel, C and Mariani, S and Escartin, S and Roiz, D and Collantes, F and Bengoa, M and Montalvo, T and Delgado, JA and Eritja, R and Lucientes, J and Albó Timor, A and Bartumeus, F and Ventura, M}, title = {Multiple invasions, Wolbachia and human-aided transport drive the genetic variability of Aedes albopictus in the Iberian Peninsula.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {20682}, pmid = {36450768}, issn = {2045-2322}, mesh = {Animals ; Humans ; *Wolbachia/genetics ; *Aedes/genetics ; Bayes Theorem ; Mosquito Vectors/genetics ; Disease Vectors ; Hydrolases ; }, abstract = {The Asian tiger mosquito, Aedes albopictus, is one of the most invasive species in the world. Native to the tropical forests of Southeast Asia, over the past 30 years it has rapidly spread throughout tropical and temperate regions of the world. Its dramatic expansion has resulted in public health concerns as a consequence of its vector competence for at least 16 viruses. Previous studies showed that Ae. albopictus spread has been facilitated by human-mediated transportation, but much remains unknown about how this has affected its genetic attributes. Here we examined the factors that contributed to shaping the current genetic constitution of Ae. albopictus in the Iberian Peninsula, where the species was first found in 2004, by combining population genetics and Bayesian modelling. We found that both mitochondrial and nuclear DNA markers showed a lack of genetic structure and the presence of worldwide dominant haplotypes, suggesting regular introductions from abroad. Mitochondrial DNA showed little genetic diversity compared to nuclear DNA, likely explained by infection with maternally transmitted bacteria of the genus Wolbachia. Multilevel models revealed that greater mosquito fluxes (estimated from commuting patterns and tiger mosquito population distribution) and spatial proximity between sampling sites were associated with lower nuclear genetic distance, suggesting that rapid short- and medium-distance dispersal is facilitated by humans through vehicular traffic. This study highlights the significant role of human transportation in shaping the genetic attributes of Ae. albopictus and promoting regional gene flow, and underscores the need for a territorially integrated surveillance across scales of this disease-carrying mosquito.}, } @article {pmid36447246, year = {2022}, author = {Dieng, MM and Augustinos, AA and Demirbas-Uzel, G and Doudoumis, V and Parker, AG and Tsiamis, G and Mach, RL and Bourtzis, K and Abd-Alla, AMM}, title = {Interactions between Glossina pallidipes salivary gland hypertrophy virus and tsetse endosymbionts in wild tsetse populations.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {447}, pmid = {36447246}, issn = {1756-3305}, mesh = {Animals ; Cytomegalovirus ; *Tsetse Flies ; *Coinfection ; *Glossinidae ; Hypertrophy ; *Infertility ; Salivary Glands ; }, abstract = {BACKGROUND: Tsetse control is considered an effective and sustainable tactic for the control of cyclically transmitted trypanosomosis in the absence of effective vaccines and inexpensive, effective drugs. The sterile insect technique (SIT) is currently used to eliminate tsetse fly populations in an area-wide integrated pest management (AW-IPM) context in Senegal. For SIT, tsetse mass rearing is a major milestone that associated microbes can influence. Tsetse flies can be infected with microorganisms, including the primary and obligate Wigglesworthia glossinidia, the commensal Sodalis glossinidius, and Wolbachia pipientis. In addition, tsetse populations often carry a pathogenic DNA virus, the Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) that hinders tsetse fertility and fecundity. Interactions between symbionts and pathogens might affect the performance of the insect host.

METHODS: In the present study, we assessed associations of GpSGHV and tsetse endosymbionts under field conditions to decipher the possible bidirectional interactions in different Glossina species. We determined the co-infection pattern of GpSGHV and Wolbachia in natural tsetse populations. We further analyzed the interaction of both Wolbachia and GpSGHV infections with Sodalis and Wigglesworthia density using qPCR.

RESULTS: The results indicated that the co-infection of GpSGHV and Wolbachia was most prevalent in Glossina austeni and Glossina morsitans morsitans, with an explicit significant negative correlation between GpSGHV and Wigglesworthia density. GpSGHV infection levels > 10[3.31] seem to be absent when Wolbachia infection is present at high density (> 10[7.36]), suggesting a potential protective role of Wolbachia against GpSGHV.

CONCLUSION: The result indicates that Wolbachia infection might interact (with an undefined mechanism) antagonistically with SGHV infection protecting tsetse fly against GpSGHV, and the interactions between the tsetse host and its associated microbes are dynamic and likely species specific; significant differences may exist between laboratory and field conditions.}, } @article {pmid36445499, year = {2022}, author = {Ramirez, P and Leavitt, JC and Gill, JJ and Mateos, M}, title = {Preliminary Characterization of Phage-Like Particles from the Male-Killing Mollicute Spiroplasma poulsonii (an Endosymbiont of Drosophila).}, journal = {Current microbiology}, volume = {80}, number = {1}, pages = {6}, pmid = {36445499}, issn = {1432-0991}, mesh = {Male ; Animals ; Drosophila ; *Bacteriophages/genetics ; Drosophila melanogaster ; *Spiroplasma/genetics ; }, abstract = {Bacteriophages are vastly abundant, diverse, and influential, but with few exceptions (e.g. the Proteobacteria genera Wolbachia and Hamiltonella), the role of phages in heritable bacteria-arthropod interactions, which are ubiquitous and diverse, remains largely unexplored. Despite prior studies documenting phage-like particles in the mollicute Spiroplasma associated with Drosophila flies, genomic sequences of such phage are lacking, and their effects on the Spiroplasma-Drosophila interaction have not been comprehensively characterized. We used a density step gradient to isolate phage-like particles from the male-killing bacterium Spiroplasma poulsonii (strains NSRO and MSRO-Br) harbored by Drosophila melanogaster. Isolated particles were subjected to DNA sequencing, assembly, and annotation. Several lines of evidence suggest that we recovered phage-like particles of similar features (shape, size, DNA content) to those previously reported in Drosophila-associated Spiroplasma strains. We recovered three ~ 19 kb phage-like contigs (two in NSRO and one in MSRO-Br) containing 21-24 open reading frames, a read-alignment pattern consistent with circular permutation, and terminal redundancy (at least in NSRO). Although our results do not allow us to distinguish whether these phage-like contigs represent infective phage-like particles capable of transmitting their DNA to new hosts, their encoding of several typical phage genes suggests that they are at least remnants of functional phage. We also recovered two smaller non-phage-like contigs encoding a known Spiroplasma toxin (Ribosome Inactivating Protein; RIP), and an insertion element, suggesting that they are packaged into particles. Substantial homology of our particle-derived contigs was found in the genome assemblies of members of the Spiroplasma poulsonii clade.}, } @article {pmid36434644, year = {2022}, author = {Vlaschenko, A and Răileanu, C and Tauchmann, O and Muzyka, D and Bohodist, V and Filatov, S and Rodenko, O and Tovstukha, I and Silaghi, C}, title = {First data on bacteria associated with bat ectoparasites collected in Kharkiv oblast, Northeastern Ukraine.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {443}, pmid = {36434644}, issn = {1756-3305}, mesh = {Animals ; Humans ; Phylogeny ; Ukraine/epidemiology ; *Argas/genetics ; *Bartonella/genetics ; Ehrlichia/genetics ; Anaplasma/genetics ; *Borrelia ; *Argasidae ; *Flea Infestations ; *Siphonaptera ; *Babesia/genetics ; }, abstract = {BACKGROUND: Bats (Mammalia: Chiroptera) serve as natural reservoirs for many zoonotic pathogens worldwide, including vector-borne pathogens. However, bat-associated parasitic arthropods and their microbiota are thus far not thoroughly described in many regions across the globe, nor is their role in the spillover of pathogens to other vertebrate species well understood. Basic epidemiological research is needed to disentangle the complex ecological interactions among bats, their specific ectoparasites and microorganisms they harbor. Some countries, such as Ukraine, are particularly data-deficient in this respect as the ectoparasitic fauna is poorly documented there and has never been screened for the presence of medically important microorganisms. Therefore, the aims of this study were to provide first data on this topic.

METHODS: A total of 239 arthropod specimens were collected from bats. They belonged to several major groups of external parasites, including soft ticks, fleas, and nycteribiid flies from six chiropteran species in Northeastern Ukraine. The ectoparasites were individually screened for the presence of DNA of Rickettsia spp., Anaplasma/Ehrlichia spp., Bartonella spp., Borrelia spp., and Babesia spp. with conventional PCRs. Positive samples were amplified at several loci, sequenced for species identification, and subjected to phylogenetic analysis.

RESULTS: Rickettsia DNA was detected exclusively in specimens of the soft tick, Carios vespertilionis (7 out of 43 or 16.3%). Sequencing and phylogenetic analysis revealed high similarity to sequences from Rickettsia parkeri and several other Rickettsia species. Bacteria from the family Anaplasmataceae were detected in all groups of the ectoparasites (51%, 122/239 samples), belonging to the genera Anaplasma, Ehrlichia, and Wolbachia. The detection of Bartonella spp. was successful only in fleas (Nycteridopsylla eusarca) and bat flies (Nycteribia koleantii, N. pedicularia), representing 12.1% (29/239) of the collected ectoparasites. No DNA of Babesia or Borrelia species was identified in the samples.

CONCLUSIONS: We report for the first time in Ukraine the molecular detection of several bacterial agents in bat ectoparasites collected from six species of bats. The data presented extend the knowledge on the distribution of ectoparasite species in bats and their involvement in potentially circulating agents pathogenic for humans and vertebrate animals.}, } @article {pmid36425974, year = {2022}, author = {Song, SL and Yong, HS and Chua, KO and Lim, PE and Eamsobhana, P}, title = {Data set on the diversity and core members of bacterial community associated with two specialist fruit flies Bactrocera melastomatos and B. umbrosa (Insecta, Tephritidae).}, journal = {Data in brief}, volume = {45}, number = {}, pages = {108727}, pmid = {36425974}, issn = {2352-3409}, abstract = {Bactrocera melastomatos Drew & Hancock and Bactrocera umbrosa (Fabricius) are fruit flies of the subfamily Dacinae under the family Tephritidae [1]. B. melastomatos occurs in India (Andaman Island), Thailand, Peninsular Malaysia, Singapore, and Indonesia (Sumatra, Kalimantan, Java) [1] while B. umbrosa is distributed from southern Thailand and Malaysia to New Guinea and New Caledonia [2]. The adult male flies of B. melastomatos are attracted to Cue lure while the adult male flies of B. umbrosa are attracted to methyl eugenol [3]. Fruit flies of Bactrocera melastomatos infest Melastomataceae while those of B. umbrosa infest Moraceae. We compare the diversity of microbiota associated with the wild adult males of these two specialist fruit flies infesting different families of host plants. Targeted 16S rRNA gene (V3-V4 region) was sequenced using the Illumina MiSeq platform. Six bacterial phyla (Actinobacteria, Armatimonadetes, Bacteroidetes, Cyanobacteria/Melainabacteria group, Firmicutes, Proteobacteria) were detected at 97% similarity clustering and 0.001% abundance filtering. Four phyla (Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria) were present in all the specimens studied. Proteobacteria was the predominant phylum in both B. melastomatos and B. umbrosa. Enterobacteriaceae was the predominant family in UM B. melastomatos and B. umbrosa, and Orbaceae was the predominant family in Awana B. melastomatos. Klebsiella was the predominant genus in B. umbrosa, Citrobacter in UM B. melastomatos, and Orbus in Awana B. melastomatos. Double Wolbachia infections were present in UM B. melastomatos. In general, the bacterial diversity and richness varied within and between the samples of B. melastomatos and B. umbrosa.}, } @article {pmid36425043, year = {2022}, author = {Nishide, Y and Sugimoto, TN and Watanabe, K and Egami, H and Kageyama, D}, title = {Genetic variations and microbiome of the poultry red mite Dermanyssus gallinae.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {1031535}, pmid = {36425043}, issn = {1664-302X}, abstract = {The poultry red mite Dermanyssus gallinae poses a significant threat to the health of hens and poultry production. A comprehensive understanding of D. gallinae is necessary to develop sustainable and efficacious control methods. Here we examined 144 D. gallinae collected from 18 poultry farms throughout the Japanese Archipelago for their genetic variations based on mitochondrial cytochrome c oxidase subunit I (COI) sequences, and microbiome variations based on amplicon sequencing of the 16S ribosomal RNA gene. According to COI sequencing, the Japanese samples were categorized into three haplogroups, which did not reflect the geographical distribution. Microbiome analyses found that the major bacteria associated with D. gallinae were Bartonella, Cardinium, Wolbachia, and Tsukamurella, with Bartonella being most predominant. Among 144 individual mites, all possessed one of the two major types of Bartonella (Bartonella sp. A), while 140 mites possessed the other type (Bartonella sp. B). The presence of the two strains of Bartonella was also confirmed by a single copy gene, rpoB. The presence of Bartonella in laid eggs suggested transovarial vertical transmission. Given that obligate blood-feeding arthropods generally require a supply of B vitamins from symbiotic bacteria, Bartonella may play an important role in mite survival. Rickettsiella, a major symbiont in European D. gallinae populations, and suggested to be an important symbiont by genomic data, was rarely found in Japanese populations. Cardinium detected from D. gallinae fell into a major clade found widely in arthropods, whereas Wolbachia detected in Japanese D. gallinae appear to be a new lineage, located at the base of Wolbachia phylogeny. Of the mitochondrial phylogeny, infection patterns of Cardinium and Wolbachia were strongly correlated, possibly suggesting one or both of the symbionts induce reproductive manipulations and increase spread in the host populations.}, } @article {pmid36417831, year = {2022}, author = {Power, RI and Šlapeta, J}, title = {Exploration of the sensitivity to macrocyclic lactones in the canine heartworm (Dirofilaria immitis) in Australia using phenotypic and genotypic approaches.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {20}, number = {}, pages = {145-158}, pmid = {36417831}, issn = {2211-3207}, mesh = {Dogs ; Animals ; *Dirofilaria immitis/genetics ; *Dirofilariasis/drug therapy/epidemiology ; Lactones/pharmacology ; *Dog Diseases/drug therapy/epidemiology ; Microfilariae/genetics ; }, abstract = {Canine heartworm disease is a potentially deadly cardiopulmonary disease caused by the mosquito-borne filarial nematode Dirofilaria immitis. In Australia, the administration of macrocyclic lactone (ML) drugs has successfully reduced the prevalence of D. immitis infection. However, the recent re-emergence of D. immitis in dogs in Queensland, Australia and the identification of ML-resistant isolates in the USA poses an important question of whether ML-resistance has emerged in this parasite in Australia. The aim of this study was to utilise phenotypic and genotypic approaches to examine the sensitivity to ML drugs in D. immitis in Australia. To do this, we surveyed 45 dogs from Queensland and New South Wales across 3 years (2019-2022) for the presence of D. immitis infection using an antigen test, quantitative Modified Knott's test, and qPCR targeting both D. immitis and the D. immitis symbiont Wolbachia. A phenotype observed by utilising sequential quantification of microfilariae for 23/45 dogs was coupled with genetic testing of filtered microfilariae for SNPs previously associated with ML-resistance in isolates from the USA. Sixteen (16/45) dogs tested positive for D. immitis infection despite reportedly receiving 'rigorous' heartworm prevention for 12 months prior to the study, according to the owners' assessment. The phenotype and genotypic assays in this study did not unequivocally demonstrate the presence of ML-resistant D. immitis in Australia. Although the failure of 16 dogs to reduce microfilaremia by >90% after ML treatment was considered a suspect phenotype of ML-resistance, no genotypic evidence was discovered using the genetic SNP analysis. The traditional quantitative Modified Knott's test can be substituted by qPCR targeting D. immitis or associated Wolbachia endosymbiont DNA for a more rapid measurement of microfilariae levels. More definitive phenotypic evidence of resistance is critically needed before the usefulness of SNPs for the detection of ML-resistance in Australia can be properly assessed.}, } @article {pmid36401143, year = {2023}, author = {Koç, N and Nalbantoğlu, S}, title = {Microbiome comparison of Dermanyssus gallinae populations from different farm rearing systems and the presence of common endosymbiotic bacteria at developmental stages.}, journal = {Parasitology research}, volume = {122}, number = {1}, pages = {227-235}, pmid = {36401143}, issn = {1432-1955}, mesh = {Animals ; Farms ; RNA, Ribosomal, 16S/genetics ; Chickens ; *Mites/genetics ; Bacteria/genetics ; Poultry ; *Trombiculidae/genetics ; *Microbiota ; *Poultry Diseases ; *Mite Infestations/veterinary ; }, abstract = {The hematophagous arthropod, Dermanyssus gallinae (Poultry red mite, PRM) can cause remarkable economic losses in the poultry industry across the globe. Although overall composition of endosymbiotic bacteria has been shown in previous studies, how farm habitats influence the microbiome remains unclear. In the present study, we compared the bacterial communities of D. gallinae populations collected from the cage and free-range farms using next-generation sequences targeting the V3-V4 hypervariable region of the 16S rRNA gene. The QIIME2 pipeline was followed in bioinformatic analyses. Proteobacteria represented a great majority of the total bacterial community of D. gallinae from both farming systems. More specifically, Bartonella-like bacteria (40.8%) and Candidatus Cardinium (21.5%) were found to be predominant genera in free-range and cage rearing systems, respectively. However, the microbiome variation based on farming systems was not statistically significant. In addition, the presence of the five common endosymbiotic bacteria (Wolbachia, Cardinium, Rickettsiella, Spiroplasma, and Schineria) was screened in different developmental stages of D. gallinae. Cardinium was detected in all developmental stages of D. gallinae. On the other hand, Wolbachia and Rickettsiella were only found in adults/nymphs, but neither in the eggs nor larvae. To our knowledge, this study provides the first microbiome comparison at genus-level in D. gallinae populations collected from different farm habitats and will contribute to the knowledge of the biology of D. gallinae.}, } @article {pmid36400975, year = {2023}, author = {Shaikevich, E and Romanov, D}, title = {Symbiotic Wolbachia bacteria in coccinellid parasitoids: genetic diversity, horizontal transfer, and recombination.}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {26}, number = {2}, pages = {269-280}, pmid = {36400975}, issn = {1618-1905}, support = {19-04-00739//Russian Foundation for Basic Research/ ; }, mesh = {Humans ; Animals ; *Wolbachia/genetics ; *Wasps/genetics/microbiology ; Phylogeny ; Symbiosis ; Recombination, Genetic ; Genetic Variation ; }, abstract = {Parasitoids, which constitute about 25% of all insects and attack arthropods of virtually all taxa, are considered the most suitable vectors for horizontal transmission of the symbiotic bacterium Wolbachia among insects. The parasitoids studied in this article develop in the larvae and pupae of ladybirds. For the first time, Wolbachia was found in parasitic wasp species of the genus Homalotylus (Hymenoptera: Encyrtidae) and from the subfamily Tetrastichinae (Hymenoptera: Eulophidae). To characterize the Wolbachia strains, six bacterial housekeeping genes were examined and compared with previously published Wolbachia gene sequences. The same bacterial strains were found in all individuals of each species of parasitic wasps collected in different places and at different times, which indicates the absence of contamination and testifies to the heritability of the symbionts in the studied chalcids. No evidence was found that the parasitic wasps were infected with Wolbachia, identical to the symbionts of their ladybirds hosts. We found one Wolbachia strain, wHom-2, which is a product of bacterial recombination from unrelated insects, including ladybirds. The lack of correspondence between the molecular phylogenies of Wolbachia strains and mitochondrial DNA of their hosts indicates horizontal transfers of Wolbachia among parasitic wasps of the genus Homalotylus and from the subfamily Tetrastichinae.}, } @article {pmid36387058, year = {2022}, author = {Ugbe, FA and Shallangwa, GA and Uzairu, A and Abdulkadir, I}, title = {Molecular docking-based virtual screening, molecular dynamic simulation, and 3-D QSAR modeling of some pyrazolopyrimidine analogs as potent anti-filarial agents.}, journal = {In silico pharmacology}, volume = {10}, number = {1}, pages = {21}, pmid = {36387058}, issn = {2193-9616}, abstract = {UNLABELLED: Lymphatic filariasis and onchocerciasis are common filarial diseases caused by filarial worms, which co-habit symbiotically with the Wolbachia organism. One good treatment method seeks Wolbachia as a drug target. Here, a computer-aided molecular docking screening and 3-D QSAR modeling were conducted on a series of Fifty-two (52) pyrazolopyrimidine derivatives against four Wolbachia receptors, including a pharmacokinetics study and Molecular Dynamic (MD) investigation, to find a more potent anti-filarial drug. The DFT approach (B3LYP with 6-31G** option) was used for the structural optimization. Five ligand-protein interaction pairs with the highest binding affinities were identified in the order; 23_7ESX (-10.2 kcal/mol) > 14_6EEZ (- 9.0) > 29_3F4R (- 8.0) > 26_6W9O (- 7.7) ≈ doxycycline_7ESX (- 7.7), with good pharmacological interaction profiles. The built 3-D QSAR model satisfied the requirement of a good model with R[2] = 0.9425, Q[2] LOO = 0.5019, SDEC = 0.1446, and F test = 98.282. The selected molecules (14, 23, 26, and 29) perfectly obeyed Lipinski's RO5 for oral bio-availability, and showed excellent ADMET properties, except 14 with positive AMES toxicity. The result of the MD simulation showed the great stability associated with the binding of 23 onto 7ESX's binding pocket with an estimated binding free energy (MM/GBSA) of - 60.6552 kcal/mol. Therefore, 23 could be recommended as a potential anti-filarial drug molecule, and/or template for the design of more prominent inhibitors.

SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40203-022-00136-y.}, } @article {pmid36376299, year = {2022}, author = {Katsuma, S and Hirota, K and Matsuda-Imai, N and Fukui, T and Muro, T and Nishino, K and Kosako, H and Shoji, K and Takanashi, H and Fujii, T and Arimura, SI and Kiuchi, T}, title = {A Wolbachia factor for male killing in lepidopteran insects.}, journal = {Nature communications}, volume = {13}, number = {1}, pages = {6764}, pmid = {36376299}, issn = {2041-1723}, mesh = {Male ; Animals ; *Wolbachia/metabolism ; *Bombyx/genetics/metabolism ; *Moths/microbiology ; Dosage Compensation, Genetic ; Insect Proteins/genetics/metabolism ; }, abstract = {Bacterial symbionts, such as Wolbachia species, can manipulate the sexual development and reproduction of their insect hosts. For example, Wolbachia infection induces male-specific death in the Asian corn borer Ostrinia furnacalis by targeting the host factor Masculinizer (Masc), an essential protein for masculinization and dosage compensation in lepidopteran insects. Here we identify a Wolbachia protein, designated Oscar, which interacts with Masc via its ankyrin repeats. Embryonic expression of Oscar inhibits Masc-induced masculinization and leads to male killing in two lepidopteran insects, O. furnacalis and the silkworm Bombyx mori. Our study identifies a mechanism by which Wolbachia induce male killing of host progeny.}, } @article {pmid36374939, year = {2022}, author = {Lozano, S and Pritts, K and Duguma, D and Fredregill, C and Connelly, R}, title = {Independent evaluation of Wolbachia infected male mosquito releases for control of Aedes aegypti in Harris County, Texas, using a Bayesian abundance estimator.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {11}, pages = {e0010907}, pmid = {36374939}, issn = {1935-2735}, support = {U01CK000512/ACL/ACL HHS/United States ; U01 CK000512/CK/NCEZID CDC HHS/United States ; }, mesh = {Animals ; Male ; *Aedes/microbiology ; Bayes Theorem ; Mosquito Vectors ; *Pest Control, Biological ; Texas ; *Wolbachia ; }, abstract = {Among disease vectors, Aedes aegypti (L.) (Diptera: Culicidae) is one of the most insidious species in the world. The disease burden created by this species has dramatically increased in the past 50 years, and during this time countries have relied on pesticides for control and prevention of viruses borne by Ae. aegypti. The small number of available insecticides with different modes of action had led to increases in insecticide resistance, thus, strategies, like the "Incompatible Insect Technique" using Wolbachia's cytoplasmic incompatibility are desirable. We evaluated the effect of releases of Wolbachia infected Ae. aegypti males on populations of wild Ae. aegypti in the metropolitan area of Houston, TX. Releases were conducted by the company MosquitoMate, Inc. To estimate mosquito population reduction, we used a mosquito abundance Bayesian hierarchical estimator that accounted for inefficient trapping. MosquitoMate previously reported a reduction of 78% for an intervention conducted in Miami, FL. In this experiment we found a reduction of 93% with 95% credibility intervals of 86% and 96% after six weeks of continual releases. A similar result was reported by Verily Life Sciences, 96% [94%, 97%], in releases made in Fresno, CA.}, } @article {pmid36367854, year = {2022}, author = {Lau, MJ and Ross, PA and Endersby-Harshman, NM and Yang, Q and Hoffmann, AA}, title = {Wolbachia inhibits ovarian formation and increases blood feeding rate in female Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {11}, pages = {e0010913}, pmid = {36367854}, issn = {1935-2735}, mesh = {Animals ; Female ; *Aedes/physiology ; Infertility ; Larva ; *Ovary/microbiology ; *Wolbachia/physiology ; }, abstract = {Wolbachia, a gram-negative endosymbiotic bacterium widespread in arthropods, is well-known for changing the reproduction of its host in ways that increase its rate of spread, but there are also costs to hosts that can reduce this. Here we investigated a novel reproductive alteration of Wolbachia wAlbB on its host Aedes aegypti, using studies on mosquito life history traits, ovarian dissection, as well as gene expression assays. We found that an extended period of the larval stage as well as the egg stage (as previously shown) can increase the proportion of Wolbachia-infected females that become infertile; an effect which was not observed in uninfected females. Infertile females had incomplete ovarian formation and also showed a higher frequency of blood feeding following a prior blood meal, indicating that they do not enter a complete gonotrophic cycle. Treatments leading to infertility also decreased the expression of genes related to reproduction, especially the vitellogenin receptor gene whose product regulates the uptake of vitellogenin (Vg) into ovaries. Our results demonstrate effects associated with the development of infertility in wAlbB-infected Ae. aegypti females with implications for Wolbachia releases. The results also have implications for the evolution of Wolbachia infections in novel hosts.}, } @article {pmid36367848, year = {2022}, author = {Lwin, MO and Ong, Z and Panchapakesan, C and Sheldenkar, A and Soh, LT and Chen, I and Li, X and Niah, W and Vasquez, K and Sim, S and Ng, LC}, title = {Influence of public hesitancy and receptivity on reactive behaviours towards releases of male Wolbachia-Aedes mosquitoes for dengue control.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {11}, pages = {e0010910}, pmid = {36367848}, issn = {1935-2735}, mesh = {Animals ; Male ; Humans ; *Wolbachia ; *Aedes ; *Dengue/prevention & control ; *Insect Bites and Stings ; Mosquito Vectors ; }, abstract = {Singapore, a highly urbanized Asian tropical country that experiences periodic dengue outbreaks, is piloting field releases of male Wolbachia-carrying Aedes aegypti mosquitoes with the aim of suppressing urban populations of the primary dengue vector Aedes aegypti. This study proposes and assesses a model to explain the roles of hesitancy and receptivity towards Project Wolbachia-Singapore in influencing reactive mosquito prevention behaviors (reactive behaviors) towards the release of Wolbachia-Aedes mosquitoes for residents living in the release sites. Interestingly, both hesitancy and receptivity predicted greater instances of reactive behaviors. The model also examines the roles of general knowledge about Wolbachia technology, perceived severity of mosquito bites, perceived density of mosquitoes, and social responsibility as predictors of hesitancy, receptivity, and reactive behaviors towards the release of Wolbachia-Aedes mosquitoes. Hesitancy towards the project mediated the effects of general knowledge, perceived severity of mosquito bites, and perceived density of mosquitoes on reactive behaviors towards the releases, although receptivity towards the project did not. Having less knowledge about Project Wolbachia-Singapore was associated with higher hesitancy towards the project and higher likelihood of performing reactive behaviors towards the releases. Individuals who perceive mosquito bites to be more severe and think that there are more mosquitoes in their living environments were also more likely to be hesitant about the project and practice reactive behaviors. However, both hesitancy and receptivity towards the project mediated the effect of social responsibility on reactive behaviors. Receptivity towards the project was driven by social responsibility, which was also associated with reduced hesitancy towards the project. Our findings suggest that, to address the hesitancy reported by a minority of participants, future outreach efforts should focus on strengthening the public's sense of social responsibility and on tailored education campaigns targeting groups with low levels of knowledge of the project.}, } @article {pmid36360289, year = {2022}, author = {da Silva, H and Oliveira, TMP and Sallum, MAM}, title = {Bacterial Community Diversity and Bacterial Interaction Network in Eight Mosquito Species.}, journal = {Genes}, volume = {13}, number = {11}, pages = {}, pmid = {36360289}, issn = {2073-4425}, mesh = {Humans ; Animals ; RNA, Ribosomal, 16S/genetics ; Brazil ; *Culicidae/genetics ; Bacteria/genetics ; *Wolbachia/genetics ; *Microbiota/genetics ; *Mercury ; }, abstract = {Mosquitoes (Diptera: Culicidae) are found widely throughout the world. Several species can transmit pathogens to humans and other vertebrates. Mosquitoes harbor great amounts of bacteria, fungi, and viruses. The bacterial composition of the microbiota of these invertebrates is associated with several factors, such as larval habitat, environment, and species. Yet little is known about bacterial interaction networks in mosquitoes. This study investigates the bacterial communities of eight species of Culicidae collected in Vale do Ribeira (Southeastern São Paulo State) and verifies the bacterial interaction network in these species. Sequences of the 16S rRNA region from 111 mosquito samples were analyzed. Bacterial interaction networks were generated from Spearman correlation values. Proteobacteria was the predominant phylum in all species. Wolbachia was the predominant genus in Haemagogus leucocelaenus. Aedes scapularis, Aedes serratus, Psorophora ferox, and Haemagogus capricornii were the species that showed a greater number of bacterial interactions. Bacterial positive interactions were found in all mosquito species, whereas negative correlations were observed in Hg. leucocelaenus, Ae. scapularis, Ae. serratus, Ps. ferox, and Hg. capricornii. All bacterial interactions with Asaia and Wolbachia were negative in Aedes mosquitoes.}, } @article {pmid36352207, year = {2023}, author = {Ueda, M and Arai, H and Masaike, K and Nakai, M and Inoue, MN}, title = {Distinct effects of three Wolbachia strains on fitness and immune traits in Homona magnanima.}, journal = {Heredity}, volume = {130}, number = {1}, pages = {22-29}, pmid = {36352207}, issn = {1365-2540}, mesh = {Animals ; Female ; Male ; *Wolbachia/genetics ; *Moths/genetics ; Reproduction ; Phenotype ; Longevity ; Symbiosis ; }, abstract = {The endosymbiotic bacterium Wolbachia occasionally increases host fitness or manipulates host reproductions to enhance vertical transmission. Multiple Wolbachia strains can coinfect the same host individual, which alters the density as well as phenotypes of the bacteria. However, the effects of Wolbachia coinfection on host fitness remain largely unknown. Here, we examined the effects of three phylogenetically distinct Wolbachia strains, wHm-a, wHm-b, and wHm-c, on host fitness by comparing non-infected, singly infected, and triply infected Homona magnanima lines within a fixed genetic background. By examining the effects of Wolbachia on host longevity, survivorship, and reproduction, we demonstrated that single infection with either wHm-b or wHm-c reduced host reproduction, but the triple infection led to the highest intrinsic growth rate. Susceptibility to the natural pathogens such as viruses and fungi was not different among the lines regardless of Wolbachia infection status. Cellular and humoral immunities were not affected by Wolbachia in females, whereas phenoloxidase activity was suppressed in males of all Wolbachia-infected lines, implying that it was a result of the mother's curse hypothesis or a strategy of Wolbachia to increase their horizontal transmission efficiency. Although how the host's genetic diversity affects the Wolbachia fitness effects is yet unknown, our findings indicated that the effects of Wolbachia are deeply influenced by infection status and that Wolbachia could change symbiotic strategy depending on host sex and transmission route.}, } @article {pmid36343219, year = {2022}, author = {Turelli, M and Katznelson, A and Ginsberg, PS}, title = {Why Wolbachia-induced cytoplasmic incompatibility is so common.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {119}, number = {47}, pages = {e2211637119}, pmid = {36343219}, issn = {1091-6490}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Female ; Humans ; *Wolbachia/genetics ; Fertility ; Cytoplasm/metabolism ; Reproduction ; Selection, Genetic ; Symbiosis ; }, abstract = {Cytoplasmic incompatibility (CI) is the most common reproductive manipulation produced by Wolbachia, obligately intracellular alphaproteobacteria that infect approximately half of all insect species. Once infection frequencies within host populations approach 10%, intense CI can drive Wolbachia to near fixation within 10 generations. However, natural selection among Wolbachia variants within individual host populations does not favor enhanced CI. Indeed, variants that do not cause CI but increase host fitness or are more reliably maternally transmitted are expected to spread if infected females remain protected from CI. Nevertheless, approximately half of analyzed Wolbachia infections cause detectable CI. Why? The frequency and persistence of CI are more plausibly explained by preferential spread to new host species (clade selection) rather than by natural selection among variants within host populations. CI-causing Wolbachia lineages preferentially spread into new host species because 1) CI increases equilibrium Wolbachia frequencies within host populations, and 2) CI-causing variants can remain at high frequencies within populations even when conditions change so that initially beneficial Wolbachia infections become harmful. An epidemiological model describing Wolbachia acquisition and loss by host species and the loss of CI-induction within Wolbachia lineages yields simple expressions for the incidence of Wolbachia infections and the fraction of those infections causing CI. Supporting a determinative role for differential interspecific spread in maintaining CI, many Wolbachia infections were recently acquired by their host species, many show evidence for contemporary spatial spread or retreat, and rapid evolution of CI-inducing loci, especially degradation, is common.}, } @article {pmid36333754, year = {2022}, author = {Cain, JL and Norris, JK and Ripley, NE and Suri, P and Finnerty, CA and Gravatte, HS and Nielsen, MK}, title = {The microbial community associated with Parascaris spp. infecting juvenile horses.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {408}, pmid = {36333754}, issn = {1756-3305}, mesh = {Humans ; Horses ; Animals ; Female ; Male ; *Ascaridoidea/genetics ; *Ascaridida Infections/veterinary ; RNA, Ribosomal, 16S/genetics ; *Horse Diseases/parasitology ; *Microbiota ; Feces/parasitology ; }, abstract = {BACKGROUND: Parasitic nematodes, including large roundworms colloquially known as ascarids, affect the health and well-being of livestock animals worldwide. The equine ascarids, Parascaris spp., are important parasites of juvenile horses and the first ascarids to develop widespread anthelmintic resistance. The microbiota has been shown to be an important factor in the fitness of many organisms, including parasitic nematodes, where endosymbiotic Wolbachia have been exploited for treatment of filariasis in humans.

METHODS: This study used short-read 16S rRNA sequences and Illumina sequencing to characterize and compare microbiota of whole worm small intestinal stages and microbiota of male and female intestines and gonads. Diversity metrics including alpha and beta diversity, and the differential abundance analyses DESeq2, ANCOM-BC, corncob, and metagenomeSeq were used for comparisons.

RESULTS: Alpha and beta diversity of whole worm microbiota did not differ significantly between groups, but Simpson alpha diversity was significantly different between female intestine (FI) and male gonad (MG) (P= 0.0018), and Shannon alpha diversity was significantly different between female and male gonads (P = 0.0130), FI and horse jejunum (HJ) (P = 0.0383), and FI and MG (P= 0.0001). Beta diversity (Fig. 2B) was significantly different between female and male gonads (P = 0.0006), male intestine (MI) and FG (P = 0.0093), and MG and FI (P = 0.0041). When comparing organs, Veillonella was differentially abundant for DESeq2 and ANCOM-BC (p < 0.0001), corncob (P = 0.0008), and metagenomeSeq (P = 0.0118), and Sarcina was differentially abundant across four methods (P < 0.0001). Finally, the microbiota of all individual Parascaris spp. specimens were compared to establish shared microbiota between groups.

CONCLUSIONS: Overall, this study provided important information regarding the Parascaris spp. microbiota and provides a first step towards determining whether the microbiota may be a viable target for future parasite control options.}, } @article {pmid36318064, year = {2022}, author = {Martinez, J and Ross, PA and Gu, X and Ant, TH and Murdochy, SM and Tong, L and da Silva Filipe, A and Hoffmann, AA and Sinkins, SP}, title = {Genomic and Phenotypic Comparisons Reveal Distinct Variants of Wolbachia Strain wAlbB.}, journal = {Applied and environmental microbiology}, volume = {88}, number = {22}, pages = {e0141222}, pmid = {36318064}, issn = {1098-5336}, support = {/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Humans ; *Wolbachia/genetics ; *Dengue Virus ; Mosquito Vectors ; *Aedes/microbiology ; *Dengue ; Genomics ; }, abstract = {The intracellular bacterium Wolbachia inhibits virus replication and is being harnessed around the world to fight mosquito-borne diseases through releases of mosquitoes carrying the symbiont. Wolbachia strains vary in their ability to invade mosquito populations and suppress viruses in part due to differences in their density within the insect and associated fitness costs. Using whole-genome sequencing, we demonstrate the existence of two variants in wAlbB, a Wolbachia strain being released in natural populations of Aedes aegypti mosquitoes. The two variants display striking differences in genome architecture and gene content. Differences in the presence/absence of 52 genes between variants include genes located in prophage regions and others potentially involved in controlling the symbiont's density. Importantly, we show that these genetic differences correlate with variation in wAlbB density and its tolerance to heat stress, suggesting that different wAlbB variants may be better suited for field deployment depending on local environmental conditions. Finally, we found that the wAlbB genome remained stable following its introduction in a Malaysian mosquito population. Our results highlight the need for further genomic and phenotypic characterization of Wolbachia strains in order to inform ongoing Wolbachia-based programs and improve the selection of optimal strains in future field interventions. IMPORTANCE Dengue is a viral disease transmitted by Aedes mosquitoes that threatens around half of the world population. Recent advances in dengue control involve the introduction of Wolbachia bacterial symbionts with antiviral properties into mosquito populations, which can lead to dramatic decreases in the incidence of the disease. In light of these promising results, there is a crucial need to better understand the factors affecting the success of such strategies, in particular the choice of Wolbachia strain for field releases and the potential for evolutionary changes. Here, we characterized two variants of a Wolbachia strain used for dengue control that differ at the genomic level and in their ability to replicate within the mosquito. We also found no evidence for the evolution of the symbiont within the 2 years following its deployment in Malaysia. Our results have implications for current and future Wolbachia-based health interventions.}, } @article {pmid36317957, year = {2023}, author = {Zhou, JC and Shang, D and Liu, SM and Zhang, C and Huo, LX and Zhang, LS and Dong, H}, title = {Wolbachia-infected Trichogramma dendrolimi is outcompeted by its uninfected counterpart in superparasitism but does not have developmental delay.}, journal = {Pest management science}, volume = {79}, number = {3}, pages = {1005-1017}, doi = {10.1002/ps.7269}, pmid = {36317957}, issn = {1526-4998}, support = {CAAS-ZDRW202108//Agricultural Science and Technology Innovation Program/ ; //Major Projects of China National Tobacco Corporation/ ; 32102275//National Natural Science Foundation of China/ ; 32172476//National Natural Science Foundation of China/ ; //Projects of Guizhou Tobacco Corporation/ ; 201936//Guizhou Tobacco Corporation/ ; 201937/WT_/Wellcome Trust/United Kingdom ; 201941/WT_/Wellcome Trust/United Kingdom ; 110202001032 (LS-01)//China National Tobacco Corporation/ ; 201937/WT_/Wellcome Trust/United Kingdom ; 201941/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Female ; Oviposition ; *Wasps/microbiology ; *Wolbachia ; }, abstract = {BACKGROUND: Wolbachia infection increases the superparasitism frequency of Trichogramma females and provides an opportunity for horizontal intraspecific transmission. However, superparasitism may lead to interstrain competition between Wolbachia-infected Trichogramma offspring and their uninfected counterparts. This study investigated the outcome of interstrain intrinsic competition between Wolbachia-infected thelytokous strain (W) and uninfected bisexual strain (B) of Trichogramma dendrolimi. To determine the developmental rate of both strains, the sizes of immature stages of T. dendrolimi offspring at different times after parasitisation were measured in single parasitism and superparasitism conditions.

RESULTS: The results reflect increased superparasitism by Wolbachia-infected females compared with uninfected females. Trichogramma females did not discriminate between host eggs previously parasitised by either B or W females. When the first oviposition was performed by B females, the B offspring outcompeted W offspring deposited later. Although when W offspring was deposited 8 h earlier than the B offspring, it gained no advantage over B offspring. Regardless of parasitism conditions, differences in the development rate between W and B offspring were not significant.

CONCLUSION: The results reconfirmed that W females presented a higher tendency for superparasitism than B females, and showed that B offspring outcompeted W offspring even when the latter was deposited 8 h earlier. The inferiority of Wolbachia-infected Trichogramma compared with their uninfected counterparts is not due to the developmental delay. This study provides insights into the effects of intrinsic competition on the control efficacy of Wolbachia-infected Trichogramma against pests in biological control programs. © 2022 Society of Chemical Industry.}, } @article {pmid36316689, year = {2022}, author = {Manvell, C and Berman, H and Callahan, B and Breitschwerdt, E and Swain, W and Ferris, K and Maggi, R and Lashnits, E}, title = {Identification of microbial taxa present in Ctenocephalides felis (cat flea) reveals widespread co-infection and associations with vector phylogeny.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {398}, pmid = {36316689}, issn = {1756-3305}, support = {T32 GM133366/GM/NIGMS NIH HHS/United States ; 1T32GM133366/NH/NIH HHS/United States ; T32OD011130/NH/NIH HHS/United States ; }, mesh = {Animals ; Cats ; *Bartonella/genetics ; Cat Diseases/parasitology ; *Coinfection ; *Ctenocephalides/microbiology ; Flea Infestations/epidemiology/veterinary ; Phylogeny ; *Rickettsia/genetics ; }, abstract = {BACKGROUND: Ctenocephalides felis, the cat flea, is the most common ectoparasite of cats and dogs worldwide. As a cause of flea allergy dermatitis and a vector for two genera of zoonotic pathogens (Bartonella and Rickettsia spp.), the effect of the C. felis microbiome on pathogen transmission and vector survival is of substantial medical importance to both human and veterinary medicine. The aim of this study was to assay the pathogenic and commensal eubacterial microbial communities of individual C. felis from multiple geographic locations and analyze these findings by location, qPCR pathogen prevalence, and flea genetic diversity.

METHODS: 16S Next Generation Sequencing (NGS) was utilized to sequence the microbiome of fleas collected from free-roaming cats, and the cox1 gene was used for flea phylogenetic analysis. NGS data were analyzed for 168 individual fleas from seven locations within the US and UK. Given inconsistency in the genera historically reported to constitute the C. felis microbiome, we utilized the decontam prevalence method followed by literature review to separate contaminants from true microbiome members.

RESULTS: NGS identified a single dominant and cosmopolitan amplicon sequence variant (ASV) from Rickettsia and Wolbachia while identifying one dominant Bartonella clarridgeiae and one dominant Bartonella henselae/Bartonella koehlerae ASV. Multiple less common ASVs from these genera were detected within restricted geographical ranges. Co-detection of two or more genera (Bartonella, Rickettsia, and/or Wolbachia) or multiple ASVs from a single genus in a single flea was common. Achromobacter, Peptoniphilus, and Rhodococcus were identified as additional candidate members of the C. felis microbiome on the basis of decontam analysis and literature review. Ctenocephalides felis phylogenetic diversity as assessed by the cox1 gene fell within currently characterized clades while identifying seven novel haplotypes. NGS sensitivity and specificity for Bartonella and Rickettsia spp. DNA detection were compared to targeted qPCR.

CONCLUSIONS: Our findings confirm the widespread coinfection of fleas with multiple bacterial genera and strains, proposing three additional microbiome members. The presence of minor Bartonella, Rickettsia, and Wolbachia ASVs was found to vary by location and flea haplotype. These findings have important implications for flea-borne pathogen transmission and control.}, } @article {pmid36314668, year = {2022}, author = {Roslan, MA and Ngui, R and Vythilingam, I and Wan Sulaiman, WY}, title = {Community surveillance of Aedes albopictus associated with Wolbachia detection in low-rise residential areas in Selangor, Malaysia.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {47}, number = {2}, pages = {142-152}, doi = {10.52707/1081-1710-47.2.142}, pmid = {36314668}, issn = {1948-7134}, mesh = {Animals ; *Wolbachia ; *Aedes ; Malaysia ; Temperature ; Polymerase Chain Reaction ; }, abstract = {The study assessed the distribution of Malaysian Ae. albopictus adults associated with Wolbachia detection in low-rise residential areas using a modified sticky ovitrap (MSO). The relationship between Ae. albopictus and climatological parameters were also determined. Fifty-two weeks of surveillance using 273 MSOs were conducted in four installation areas of eleven sampling sites. Specimens were subjected to PCR using wsp-specific primers for Wolbachia detection. The relationship between climatological parameters and Ae. albopictus captured were analyzed using Spearman rank correlation coefficient test. The majority of Ae. albopictus were captured in residential houses (87%), followed by playgrounds or parks (11.5%), guardhouses (1%), and community halls (0.5%). Most of the specimens (92%) were superinfected with wAlbA and wAlbB strains. A positive correlation with no significant association was found for rainfall (r = 0.015, P = 0.072), relative humidity (r = 0.005, P = 0.526), minimum temperature (r = 0.005, P = 0.516), and mean temperature (r = 0.003, P = 0.689). MSO effectively captured a high number of Ae. albopictus that was determined to be the predominant mosquito species found in low-rise residential areas. The adult collection is not only influenced by climatological parameters but also by other factors, including environmental conditions and general sanitation status.}, } @article {pmid36313820, year = {2022}, author = {Ryabinin, AS and Shishkina, OD and Ilinsky, YY and Bykov, RA}, title = {Rare Wolbachia genotypes in laboratory Drosophila melanogaster strains.}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {26}, number = {6}, pages = {553-559}, doi = {10.18699/VJGB-22-67}, pmid = {36313820}, issn = {2500-0462}, abstract = {Symbiotic bacteria of the genus Wolbachia are widespread in Drosophila melanogaster populations. Based on the polymorphism of the Wolbachia genome, the symbionts' diversity in D. melanogaster is presented by two groups: MEL (wMel, wMel2, wMel3 and wMel4) and CS (wMelCS and wMelCS2). The wMel genotype is predominant in natural D. melanogaster populations and is distributed all over the world. The CS genotypes, on the other hand, are of particular interest because it is unclear how they are maintained in the fruit f ly populations since they should have been eliminated from them due to their low frequency and genetic drift or been replaced by the wMel genotype. However, this is not what is really observed, which means these genotypes are supported by selection. It is known that the wMelPlus strain of the wMelCS genotype can increase the lifespan of infected f lies at high temperatures. The same genotype also increases the intensity of dopamine metabolism in Drosophila compared to the MEL-group genotypes. In the present study, we searched for the rare Wolbachia wMelCS and wMelCS2 genotypes, as well as for new genotypes in wild-type D. melanogaster strains and in several mutant laboratory strains. The symbiont was found in all populations, in 200 out of 385 wild-type strains and in 83 out of 170 mutant strains. Wolbachia diversity in D. melanogaster wild-type strains was represented by the wMel, wMelCS and wMelCS2 genotypes. More than 90 % of the infected strains carried wMel; 9 %, wMelCS2; and only two strains were found to carry wMelCS. No new Wolbachia genotypes were found. The northernmost point reported for the wMelCS2 genotype was Izhevsk city (Udmurtia, Russia). For the f irst time the wMelCS2 genotype was detected in D. melanogaster from the Sakhalin Island, and wMelCS, in the f lies from Nalchik (the North Caucasus). A comparison of Wolbachia genetic diversity between the wild-type laboratory strains and previously obtained data on mutant laboratory strains demonstrated differences in the frequencies of rare CS genotypes, which were more prevalent in mutant strains, apparently due to the breeding history of these Drosophila strains.}, } @article {pmid36304111, year = {2022}, author = {Ritchie, IT and Needles, KT and Leigh, BA and Kaur, R and Bordenstein, SR}, title = {Transgenic cytoplasmic incompatibility persists across age and temperature variation in Drosophila melanogaster.}, journal = {iScience}, volume = {25}, number = {11}, pages = {105327}, pmid = {36304111}, issn = {2589-0042}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, abstract = {Environmental stressors can impact the basic biology and applications of host-microbe symbioses. For example, Wolbachia symbiont densities and cytoplasmic incompatibility (CI) levels can decline in response to extreme temperatures and host aging. To investigate whether transgenic expression of CI-causing cif genes overcomes the environmental sensitivity of CI, we exposed transgenic male flies to low and high temperatures as well as aging treatments. Our results indicate that transgenic cif expression induces nearly complete CI regardless of temperature and aging, despite severe weakening of Wolbachia-based wild-type CI. Strong CI levels correlate with higher levels of cif transgene expression in young males. Altogether, our results highlight that transgenic CI persists against common environmental pressures and may be relevant for future control applications involving the cifA and cifB transgenes.}, } @article {pmid36303000, year = {2022}, author = {Kache, PA and Santos-Vega, M and Stewart-Ibarra, AM and Cook, EM and Seto, KC and Diuk-Wasser, MA}, title = {Bridging landscape ecology and urban science to respond to the rising threat of mosquito-borne diseases.}, journal = {Nature ecology & evolution}, volume = {6}, number = {11}, pages = {1601-1616}, pmid = {36303000}, issn = {2397-334X}, support = {U01 CK000509/CK/NCEZID CDC HHS/United States ; }, mesh = {Animals ; Humans ; Mosquito Vectors ; *Aedes ; Ecology ; Urbanization ; *Vector Borne Diseases/prevention & control ; }, abstract = {The prevalence of diseases borne by mosquitoes, particularly in the genus Aedes, is rising worldwide. This has been attributed, in part, to the dramatic rates of contemporary urbanization. While Aedes-borne disease risk varies within and between cities, few investigations use urban science-based approaches to examine how city structure and function contribute to vector or pathogen introduction and maintenance. Here, we integrate theories from complex adaptive systems, landscape ecology and urban geography to develop an urban systems framework for understanding Aedes-borne diseases. The framework establishes that cities comprise hierarchically structured patches of different land uses and characteristics. Properties of the patches (that is, composition) determine localized disease risk, while configuration and connectivity drive emergent patterns of pathogen spread. Complexity is added by incorporating individual and collective human social structures, considering how feedbacks among social actors and with the landscape drive risk and transmission. We discuss how these concepts apply to case studies of Aedes-borne disease from around the world. Ultimately, the framework strengthens existing theoretical and mixed qualitative-quantitative approaches, and advances considerations of how interventions including urban planning (for example, piped water provisioning) and emerging vector control strategies (for example, Wolbachia-infected mosquitoes) can be implemented to prevent and control the rising threat of Aedes-borne diseases.}, } @article {pmid36299486, year = {2022}, author = {Hargitai, D and Kenéz, L and Al-Lami, M and Szenczi, G and Lőrincz, P and Juhász, G}, title = {Autophagy controls Wolbachia infection upon bacterial damage and in aging Drosophila.}, journal = {Frontiers in cell and developmental biology}, volume = {10}, number = {}, pages = {976882}, pmid = {36299486}, issn = {2296-634X}, abstract = {Autophagy is a conserved catabolic process in eukaryotic cells that degrades intracellular components in lysosomes, often in an organelle-specific selective manner (mitophagy, ERphagy, etc). Cells also use autophagy as a defense mechanism, eliminating intracellular pathogens via selective degradation known as xenophagy. Wolbachia pipientis is a Gram-negative intracellular bacterium, which is one of the most common parasites on Earth affecting approximately half of terrestrial arthropods. Interestingly, infection grants the host resistance against other pathogens and modulates lifespan, so this bacterium resembles an endosymbiont. Here we demonstrate that Drosophila somatic cells normally degrade a subset of these bacterial cells, and autophagy is required for selective elimination of Wolbachia upon antibiotic damage. In line with these, Wolbachia overpopulates in autophagy-compromised animals during aging while its presence fails to affect host lifespan unlike in case of control flies. The autophagic degradation of Wolbachia thus represents a novel antibacterial mechanism that controls the propagation of this unique bacterium, behaving both as parasite and endosymbiont at the same time.}, } @article {pmid36296267, year = {2022}, author = {Ibrahim, YE and Paredes-Montero, JR and Al-Saleh, MA and Widyawan, A and He, R and El Komy, MH and Al Dhafer, HM and Kitchen, N and Gang, DR and Brown, JK}, title = {Characterization of the Asian Citrus Psyllid-'Candidatus Liberibacter Asiaticus' Pathosystem in Saudi Arabia Reveals Two Predominant CLas Lineages and One Asian Citrus Psyllid Vector Haplotype.}, journal = {Microorganisms}, volume = {10}, number = {10}, pages = {}, pmid = {36296267}, issn = {2076-2607}, support = {2018-70016-27411//USDA-NIFA CDRE/ ; 14-BIO-627-02//National Plan for Science, Technology, and Innovation (MAARIFAH), King Abdul-Aziz City for Science and Technology, Kingdom of Saudi Arabia/ ; }, abstract = {In Saudi Arabia (SA), the citrus greening disease is caused by 'Candidatus Liberibacter asiaticus' (CLas) transmitted by the Asian citrus psyllid (ACP) Diaphorina citri. The origin and route(s) of the ACP-CLas pathosystem invasion in SA have not been studied. Adult ACP were collected from citrus trees in SA and differentiated by analysis of the mitochondrial cytochrome oxidase I (mtCOI) and nuclear copper transporting protein (atox1) genes. A phylogenetic analysis of the Wolbachia spp. surface protein (wsp) gene was used to identify the ACP-associated Wolbachia spp. A phylogenetic analysis of the atox1 and mtCOI gene sequences revealed one predominant ACP haplotype most closely related to the Indian subcontinent founder populations. The detection and identification of CLas in citrus trees were carried out by polymerase chain reaction (PCR) amplification and sequencing of the 16S rDNA gene. The CLas-integrated prophage genomes were sequenced, annotated, and used to differentiate CLas populations. The ML and ASTRAL trees reconstructed with prophages type 1 and 2 genome sequences, separately and concatenated, resolved two major lineages, CLas-1 and -2. The CLas-1 clade, reported here for the first time, consisted of isolates from SA isolates and Pakistan. The CLas-2 sequences formed two groups, CLas-2-1 and -2-2, previously the 'Asiatic' and 'Floridian' strains, respectively. Members of CLas-2-1 originated from Southeast Asia, the USA, and other worldwide locations, while CLas-2-2 was identified only in Florida. This study provides the first snapshot into the status of the ACP-CLas pathosystem in SA. In addition, the results provide new insights into the pathosystem coevolution and global invasion histories of two ACP-CLas lineages with a predicted center of origin in South and Southeast Asia, respectively.}, } @article {pmid36296233, year = {2022}, author = {Ngnindji-Youdje, Y and Diarra, AZ and Lontsi-Demano, M and Tchuinkam, T and Parola, P}, title = {Detection of Tick-Borne Pathogens in Ticks from Cattle in Western Highlands of Cameroon.}, journal = {Microorganisms}, volume = {10}, number = {10}, pages = {}, pmid = {36296233}, issn = {2076-2607}, support = {ANR-10-IAHU-03//Méditerranée Infection Foundation/ ; }, abstract = {This study aimed to detect and identify microorganisms in ticks collected in the Western Highlands of Cameroon. Quantitative real-time and standard PCR assays, coupled with sequencing, were used. A total of 944 ticks collected from cattle in five distinct sites in Cameroon were selected for the analyses. They belonged to five genera (Amblyomma, Hyalomma, Rhipicephalus, Haemaphysalis, and Ixodes) and twelve species. Real-time PCR revealed that 23% (n = 218) of the ticks were positive for Rickettsia spp., 15% (n = 141) for bacteria of the Anaplasmataceae family, 3% (n = 29) for Piroplasmida, 0.5% (n = 5) for Coxiella burnetii, 0.4% (n = 4) for Borrelia spp., and 0.2% (n = 2) for Bartonella spp. The co-infection rate (3.4%, n = 32) involved mainly Rickettsia spp. and Anaplasmataceae. Of the Rickettsia spp. positive ticks, the targeted PCR and sequencing yielded Rickettsia africae (78.9%), Rickettsia aeschlimannii (6.4%), Rickettsia massiliae (7.8%), Candidatus Rickettsia barbariae (0.9%), and Rickettsia sp. (0.9%). Anaplasmataceae included Anaplasma marginale (4.3%), Anaplasma platys (1.4%), Anaplasma centrale (0.7%), Ehrlichia ruminantium (0.7%), Wolbachia sp., Candidatus Ehrlichia rustica (13.5%), Candidatus Ehrlichia urmitei (7%), and an uncultured Ehrlichia sp. (4.2%). Borrelia theileri was identified in one Rhipicephalus microplus tick. Unfortunately, Piroplasmida could not be identified to the species level. This study demonstrates that in Cameroon, ticks harbour a wide variety of microorganisms and present a risk of zoonotic diseases.}, } @article {pmid36292897, year = {2022}, author = {Pagendam, D and Elfekih, S and Nassar, MS and Nelson, S and Almalik, AM and Tawfik, EA and Al-Fageeh, MB and Hoffmann, AA}, title = {Spatio-Temporal Modelling Informing Wolbachia Replacement Releases in a Low Rainfall Climate.}, journal = {Insects}, volume = {13}, number = {10}, pages = {}, pmid = {36292897}, issn = {2075-4450}, support = {ETSC&KACST-CSIRO-2018-12-30-21//KACST-CSIRO collaborative research agreement/ ; 132412//National Health and Medical Research Council/ ; 1118640//National Health and Medical Research Council/ ; }, abstract = {Releases of Aedes aegypti carrying Wolbachia bacteria are known to suppress arbovirus transmission and reduce the incidence of vector-borne diseases. In planning for Wolbachia releases in the arid environment of Jeddah, Saudi Arabia, we collected entomological data with ovitraps across a 7-month period in four locations. Herein, we show that mosquito presence in basements does not differ from that of non-basement areas of buildings. In modelling mosquito presence across the study sites, we found the spatial structure to be statistically significant in one of the four sites, while a significant spatial structure was found for egg production data across three of the four sites. The length scales of the spatial covariance functions fitted to the egg production data ranged from 143 m to 574 m, indicating that high productivity regions can be extensive in size. Rank-correlation analyses indicated that mosquito presence tended to persist from the dry to wet season, but that egg production ranks at locations could reverse. The data suggest that, in Jeddah, the quality of the local environment for breeding can vary over time. The data support the feasibility of dry season releases but with release numbers needing to be flexible depending on local rates of invasion.}, } @article {pmid36292872, year = {2022}, author = {Buchori, D and Mawan, A and Nurhayati, I and Aryati, A and Kusnanto, H and Hadi, UK}, title = {Risk Assessment on the Release of Wolbachia-Infected Aedes aegypti in Yogyakarta, Indonesia.}, journal = {Insects}, volume = {13}, number = {10}, pages = {}, pmid = {36292872}, issn = {2075-4450}, support = {28-Mei-2016//Yayasan Tahija/ ; }, abstract = {Wolbachia-infected Aedes aegypti is the latest technology that was developed to eliminate dengue fever. The Ministry of Research and Technology of the Republic of Indonesia (Kemenristekdikti) established an expert group to identify future potential risks that may occur over a period of 30 years associated with the release of Wolbachia-infected Ae. aegypti. The risk assessment consisted of identifying different hazards that may have impacts on humans and the environment. From the consensus among the experts, there were 56 hazards identified and categorized into 4 components, namely, ecological matters, efficacy in mosquito management, economic and sociocultural issues, and public health standards. There were 19 hazards in the ecological group. The overall likelihood in the ecology of the mosquito is very low (0.05), with moderate consequence (0.74), which resulted in negligible risk. For the efficacy in mosquito management group, there were 12 hazards that resulted in very low likelihood (0.11) with high consequence (0.85). The overall risk for mosquito management efficacy was very low (0.09). There were 14 hazards identified in the public health standard with very low likelihood (0.07), moderate consequence (0.50) and negligible risk (0.04). Lastly, 13 hazards were identified in the economic and sociocultural group with low likelihood (0.01) but of moderate consequence (0.5), which resulted in a very low risk (0.09). The risk severity level of the four components leading to the endpoint risk of "cause more harm" due to releasing Wolbachia-infected Ae. aegypti is negligible (0.01).}, } @article {pmid36271174, year = {2022}, author = {Fallon, AM}, title = {Mitotically inactivated mosquito cells support robust Wolbachia infection and replication.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {58}, number = {9}, pages = {780-787}, pmid = {36271174}, issn = {1543-706X}, mesh = {Animals ; *Wolbachia ; *Aedes ; Cell Line ; Mammals ; }, abstract = {Wolbachia is an obligate intracellular bacterium that infects many species of insects, and has been of particular interest in recent efforts to reduce disease transmission by mosquitoes. Two aspects of Wolbachia biology underlie its applications for insect control: first, the bacterium behaves as a natural gene drive agent and, second, when introduced into mosquitoes that do not harbor Wolbachia in nature, infection reduces survival of pathogens. These properties support efforts to explore the basic biology of Wolbachia in insect cell lines, which can produce sufficient infectious material for microbiological studies and microinjection into novel hosts. When introduced into naïve C7-10 Aedes albopictus mosquito cells, the yield of Wolbachia strain wStri improves, roughly in proportion to the size of the inoculum, as exponential growth of the host cell ceases. Wolbachia yields also increase when persistently infected C/wStri1 cells or naive, newly infected cells are treated with 20-hydroxyecdysone (20E), which inhibits growth in the G1 phase of the cell cycle. These observations suggest that Wolbachia infection and replication are independent of exponential growth and mitosis of host cells. To explore yields of infectious bacteria in cells arrested prior to infection, I tested host cells pre-treated with mitomycin C, an agent that crosslinks DNA and prevents cell division that is used to produce "feeder layers" with mammalian cells. Yields of wStri per plate increased by about 50-fold relative to exponentially growing cells, and the multiplicity of infection necessary for a robust infection was reduced to a single bacterium per cell. These results suggest that Wolbachia infection and replication are supported by mitotically arrested cells and provide new insights into biological processes that influence maintenance of a widespread obligate intracellular bacterium.}, } @article {pmid36270115, year = {2022}, author = {Araújo, IM and Cordeiro, MD and Soares, RFP and Guterres, A and Sanavria, A and Baêta, BA and da Fonseca, AH}, title = {Survey of bacterial and protozoan agents in ticks and fleas found on wild animals in the state of Rio de Janeiro, Brazil.}, journal = {Ticks and tick-borne diseases}, volume = {13}, number = {6}, pages = {102037}, doi = {10.1016/j.ttbdis.2022.102037}, pmid = {36270115}, issn = {1877-9603}, abstract = {This study evaluates the presence of bacterial and protozoan agents in ticks and fleas found on wild animals in the state of Rio de Janeiro, Brazil. These ectoparasites were collected on mammal species Hydrochoerus hydrochaeris, Tapirus terrestris, Dicotyles tajacu, Didelphis aurita, Cuniculus paca, Cerdocyon thous, and Coendou prehensilis, and on the terrestrial bird Dromaius novaehollandiae. Ticks and fleas were identified morphologically using specific taxonomic keys. A total of 396 ticks and 54 fleas were tested via polymerase chain reaction (PCR) for the presence of Rickettsia spp., Borrelia spp., microorganisms of the order Piroplasmida and Anaplasmataceae family. This total is distributed among nine tick species of the genus Amblyomma and one flea species. Rickettsia bellii was detected in Amblyomma dubitatum and Amblyomma pacae; Rickettsia sp. strain AL was found in Amblyomma longirostre; Rickettsia parkeri strain Atlantic rainforest was found in Amblyomma ovale; and "Candidatus Rickettsia senegalensis" and Rickettsia felis were detected in Ctenocephalides felis felis. Wolbachia sp. was detected in C. f. felis, and Borrelia sp. was detected in Amblyomma calcaratum (here named Borrelia sp. strain Acalc110). All tested samples were negative for Ehrlichia spp. and microorganisms of the Piroplasmida order. This study detected a new bacterial strain, Borrelia sp. strain Acalc 110 (which is genetically close to B. miyamotoi and B. venezuelensis) and the Rickettsia sp. strain 19P, which is 100% similar to "Ca. R. senegalensis", a bacterium recently discovered and now being reported for the first time in Brazil.}, } @article {pmid36250862, year = {2022}, author = {Huggins, LG and Colella, V and Atapattu, U and Koehler, AV and Traub, RJ}, title = {Nanopore Sequencing Using the Full-Length 16S rRNA Gene for Detection of Blood-Borne Bacteria in Dogs Reveals a Novel Species of Hemotropic Mycoplasma.}, journal = {Microbiology spectrum}, volume = {10}, number = {6}, pages = {e0308822}, pmid = {36250862}, issn = {2165-0497}, mesh = {Animals ; Dogs ; *Dog Diseases/diagnosis/epidemiology/microbiology ; Genes, rRNA ; High-Throughput Nucleotide Sequencing ; *Mycoplasma/classification/genetics ; *Nanopore Sequencing ; RNA, Ribosomal, 16S/genetics ; *Blood-Borne Pathogens/classification ; }, abstract = {Dogs across the globe are afflicted by diverse blood- and vector-borne bacteria (VBB), many of which cause severe disease and can be fatal. Diagnosis of VBB infections can be challenging due to the low concentration of bacteria in the blood, the frequent occurrence of coinfections, and the wide range of known, emerging, and potentially novel VBB species encounterable. Therefore, there is a need for diagnostics that address these challenges by being both sensitive and capable of detecting all VBB simultaneously. We detail the first employment of a nanopore-based sequencing methodology conducted on the Oxford Nanopore Technologies (ONT) MinION device to accurately elucidate the "hemobacteriome" from canine blood through sequencing of the full-length 16S rRNA gene. We detected a diverse range of important canine VBB, including Ehrlichia canis, Anaplasma platys, Mycoplasma haemocanis, Bartonella clarridgeiae, "Candidatus Mycoplasma haematoparvum", a novel species of hemotropic mycoplasma, and Wolbachia endosymbionts of filarial worms, indicative of filariasis. Our nanopore-based protocol was equivalent in sensitivity to both quantitative PCR (qPCR) and Illumina sequencing when benchmarked against these methods, achieving high agreement as defined by the kappa statistics (k > 0.81) for three key VBB. Utilizing the ability of the ONT' MinION device to sequence long read lengths provides an excellent alternative diagnostic method by which the hemobacteriome can be accurately characterized to the species level in a way previously unachievable using short reads. We envision our method to be translatable to multiple contexts, such as the detection of VBB in other vertebrate hosts, including humans, while the small size of the MinION device is highly amenable to field use. IMPORTANCE Blood- and vector-borne bacteria (VBB) can cause severe pathology and even be lethal for dogs in many regions across the globe. Accurate characterization of all the bacterial pathogens infecting a canine host is critical, as coinfections are common and emerging and novel pathogens that may go undetected by traditional diagnostics frequently arise. Deep sequencing using devices from Oxford Nanopore Technologies (ONT) provides a solution, as the long read lengths achievable provide species-level taxonomic identification of pathogens that previous short-read technologies could not accomplish. We developed a protocol using ONT' MinION sequencer to accurately detect and classify a wide spectrum of VBB from canine blood at a sensitivity comparable to that of regularly used diagnostics, such as qPCR. This protocol demonstrates great potential for use in biosurveillance and biosecurity operations for the detection of VBB in a range of vertebrate hosts, while the MinION sequencer's portability allows this method to be used easily in the field.}, } @article {pmid36247321, year = {2022}, author = {Tomaru, M and Takano-Shimizu-Kouno, T and Wakada, H}, title = {No Wolbachia infection was detected in Drosophila elegans collected from the wild in the Ryukyu Islands, Japan.}, journal = {microPublication biology}, volume = {2022}, number = {}, pages = {}, pmid = {36247321}, issn = {2578-9430}, abstract = {Flower breeding, tropical and subtropical Drosophila elegans is distributed in the Ryukyu Islands and Taiwan (black morph) and in southern China, Philippines, Indonesia, and New Guinea (brown morph). Although reproductive and behavioral manipulations by Wolbachia are reported in many insect taxa, Wolbachia infection in D. elegans is unclear. There is only a report of no Wolbachia detected in a laboratory strain of brown morph. This PCR diagnosis study revealed no Wolbachia infection in D. elegans males collected from the wild in the Ryukyu Islands. We concluded that D. elegans black morph in the Ryukyu Islands is not infected with Wolbachia .}, } @article {pmid36244506, year = {2022}, author = {Djoukzoumka, S and Mahamat Hassane, H and Khan Payne, V and Ibrahim, MAM and Tagueu Kanté, S and Mouliom Mfopit, Y and Berger, P and Kelm, S and Simo, G}, title = {Sodalis glossinidius and Wolbachia infections in wild population of Glossina morsitans submorsitans caught in the area of Lake Iro in the south of Chad.}, journal = {Journal of invertebrate pathology}, volume = {195}, number = {}, pages = {107835}, doi = {10.1016/j.jip.2022.107835}, pmid = {36244506}, issn = {1096-0805}, mesh = {Animals ; *Tsetse Flies/microbiology ; *Wolbachia ; Lakes ; Chad ; *Trypanosoma/genetics ; Symbiosis ; }, abstract = {Investigations on the bacterial fauna and their association with trypanosome infections in tsetse fly have revealed contrasting results. This study aimed to detect Wolbachia and S. glossinidius in wild populations of G. m. submorsistans and subsequently, understand the influence that these bacteria may have on the vectorial competence of this tsetse species. Tsetse flies were captured in the area of Lake Iro in the south of Chad using biconical traps. After DNA extraction from each tsetse fly, Sodalis glossinidius and Wolbachia were detected using specific primers. Sodalis glossinidius and Wolbachia infection rates were compared and association studies involving trypanosome infections and S. glossinidius or Wolbachia were performed. From 345 G. m. submorsitans analyzed, 9.0% and 14.5% were respectively infected with S. glossinidius and Wolbachia. Only 2.31% of all tsetse flies were co-infected by the 2 bacteria. Of all trypanosome-infected flies, 7.1% and 9.8% harbored, respectively, S. glossinidius and Wolbachia. No association was observed between Wolbachia and trypanosome infections while a significant association (r = 4.992; P = 0.025) was found between S. glossinidius and the presence of trypanosomes. A significant association (r = 3.147; P = 0.043) was also observed between S. glossinidius and T. simiae; and none with T. congolense or T. godfreyi. This study revealed S. glossinidius and Wolbachia in G. m. submorsitans of the area of lake Iro. It showed that co-infections between Wolbachia and S. glossinidius are rare in wild populations of G. m. submorsitans and that the tripartite associations vary according to trypanosome species as well as symbiotic mricroorganisms.}, } @article {pmid36227923, year = {2022}, author = {Hugo, LE and Rašić, G and Maynard, AJ and Ambrose, L and Liddington, C and Thomas, CJE and Nath, NS and Graham, M and Winterford, C and Wimalasiri-Yapa, BMCR and Xi, Z and Beebe, NW and Devine, GJ}, title = {Wolbachia wAlbB inhibit dengue and Zika infection in the mosquito Aedes aegypti with an Australian background.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {10}, pages = {e0010786}, pmid = {36227923}, issn = {1935-2735}, mesh = {*Aedes ; Animals ; Australia ; DNA ; *Dengue/prevention & control ; *Dengue Virus/physiology ; Humans ; *Insecticides ; Male ; Mosquito Vectors ; *Wolbachia/physiology ; *Zika Virus/genetics ; *Zika Virus Infection/prevention & control ; }, abstract = {Biological control of mosquito vectors using the endosymbiotic bacteria Wolbachia is an emerging strategy for the management of human arboviral diseases. We recently described the development of a strain of Aedes aegypti infected with the Wolbachia strain wAlbB (referred to as the wAlbB2-F4 strain) through simple backcrossing of wild type Australian mosquitoes with a wAlbB infected Ae. aegypti strain from the USA. Field releases of male wAlbB2-F4 mosquitoes resulted in the successful suppression of wild populations of mosquitoes in the trial sites by exploiting the strain's Wolbachia-induced cytoplasmic incompatibility. We now demonstrate that the strain is resistant to infection by dengue and Zika viruses and is genetically similar to endemic Queensland populations. There was a fourfold reduction in the proportion of wAlbB2-F4 mosquitoes that became infected following a blood meal containing dengue 2 virus (16.7%) compared to wild type mosquitoes (69.2%) and a 6-7 fold reduction in the proportion of wAlbB2-F4 mosquitoes producing virus in saliva following a blood meal containing an epidemic strain of Zika virus (8.7% in comparison to 58.3% in wild type mosquitoes). Restriction-site Associated DNA (RAD) sequencing revealed that wAlbB2-F4 mosquitoes have > 98% Australian ancestry, confirming the successful introduction of the wAlbB2 infection into the Australian genomic background through backcrossing. Genotypic and phenotypic analyses showed the wAlbB2-F4 strain retains the insecticide susceptible phenotype and genotype of native Australian mosquitoes. We demonstrate that the Wolbachia wAlbB2-F4, in addition to being suitable for population suppression programs, can also be effective in population replacement programs given its inhibition of virus infection in mosquitoes. The ease at which a target mosquito population can be transfected with wAlbB2, while retaining the genotypes and phenotypes of the target population, shows the utility of this strain for controlling the Ae. aegypti mosquitoes and the pathogens they transmit.}, } @article {pmid36214563, year = {2022}, author = {Zhou, JC and Zhao, X and Huo, LX and Shang, D and Dong, H and Zhang, LS}, title = {Wolbachia-Driven Memory Loss in a Parasitic Wasp Increases Superparasitism to Enhance Horizontal Transmission.}, journal = {mBio}, volume = {13}, number = {6}, pages = {e0236222}, pmid = {36214563}, issn = {2150-7511}, mesh = {Humans ; Animals ; Female ; *Wasps/microbiology ; *Wolbachia/genetics ; In Situ Hybridization, Fluorescence ; Larva ; Memory Disorders ; }, abstract = {Horizontal transmission of the endosymbiont, Wolbachia, may occur during superparasitism when parasitoid females deposit a second clutch of eggs on a host. Wolbachia may increase the superparasitism tendency of Trichogramma wasps by depriving their memory. To test this hypothesis, we investigated the effects of conditioning experience and memory inhibitors (actinomycin D [ACD] and anisomycin [ANI]) on memory capacity, and expressions of memory-related genes (CREB1 and PKA), and superparasitism frequency of Wolbachia-infected (TDW) and uninfected (TD) lines of Trichogramma dendrolimi after conditioning with lemon or peppermint odor. We detected the presence of Wolbachia in eggs, larvae, pre-pupae, pupae, and adults of Trichogramma by using fluorescence in situ hybridization. The results showed that TDW females had a more reduced memory capacity than TD females after conditioning. Compared with TD females, TDW females showed a higher proportion of superparasitism and a downregulation of CREB1 and PKA genes after conditioning. TD females fed ACD or ANI showed a higher tendency for superparasitism and a downregulation of CREB1 and PKA, along with memory loss after conditioning than TD females fed honey solution only. The presence of Wolbachia was detected in the anterior region of the larva, pre-pupa, and pupa, but was not found in the head of the adult. The results provide evidence of host behavioral manipulation of Wolbachia by depriving memory of host Trichogramma wasps based on Poulin' s criteria. These host behavioral changes led by Wolbachia may be caused by the virulence of Wolbachia on the nervous system of the host. IMPORTANCE The endosymbiotic bacteria, Wolbachia, live widely within cells of arthropods. Wolbachia are not only transmitted vertically from host mother to offspring, but are also transmitted horizontally among host individuals. Horizontal transmission is expected to occur during superparasitism when host parasitoid females deposit a clutch of eggs on a host previously parasitized by the same parasitoid species. Thus, a question is proposed regarding whether superparasitism behavior is a behavior modification induced by the symbiont to favor symbiont transmission. This study highlights behavioral mechanisms of Wolbachia-induced superparasitism in Trichogramma wasps and the manipulation of symbionts on host parasitoids.}, } @article {pmid36205550, year = {2023}, author = {Ant, TH and Mancini, MV and McNamara, CJ and Rainey, SM and Sinkins, SP}, title = {Wolbachia-Virus interactions and arbovirus control through population replacement in mosquitoes.}, journal = {Pathogens and global health}, volume = {117}, number = {3}, pages = {245-258}, pmid = {36205550}, issn = {2047-7732}, mesh = {Animals ; Humans ; *Arboviruses/physiology ; *Wolbachia ; *Dengue Virus/physiology ; Mosquito Vectors ; *Aedes ; *Zika Virus ; *Zika Virus Infection ; *Dengue/prevention & control ; Population Dynamics ; }, abstract = {Following transfer into the primary arbovirus vector Aedes aegypti, several strains of the intracellular bacterium Wolbachia have been shown to inhibit the transmission of dengue, Zika, and chikungunya viruses, important human pathogens that cause significant morbidity and mortality worldwide. In addition to pathogen inhibition, many Wolbachia strains manipulate host reproduction, resulting in an invasive capacity of the bacterium in insect populations. This has led to the deployment of Wolbachia as a dengue control tool, and trials have reported significant reductions in transmission in release areas. Here, we discuss the possible mechanisms of Wolbachia-virus inhibition and the implications for long-term success of dengue control. We also consider the evidence presented in several reports that Wolbachia may cause an enhancement of replication of certain viruses under particular conditions, and conclude that these should not cause any concerns with respect to the application of Wolbachia to arbovirus control.}, } @article {pmid36201377, year = {2022}, author = {Kiefer, JST and Schmidt, G and Krüsemer, R and Kaltenpoth, M and Engl, T}, title = {Wolbachia causes cytoplasmic incompatibility but not male-killing in a grain pest beetle.}, journal = {Molecular ecology}, volume = {31}, number = {24}, pages = {6570-6587}, doi = {10.1111/mec.16717}, pmid = {36201377}, issn = {1365-294X}, mesh = {Animals ; Male ; Female ; *Wolbachia/genetics ; *Coleoptera/genetics/microbiology ; Drosophila melanogaster/genetics ; Phylogeny ; Cytoplasm/genetics/microbiology ; Symbiosis/genetics ; }, abstract = {The endosymbiotic Wolbachia is one of the most common intracellular bacteria known in arthropods and nematodes. Its ability for reproductive manipulation can cause unequal inheritance to male and female offspring, allowing the manipulator to spread, but potentially also impact the evolutionary dynamics of infected hosts. Estimated to be present in up to 66% of insect species, little is known about the phenotypic impact of Wolbachia within the order Coleoptera. Here, we describe the reproductive manipulation by the Wolbachia strain wSur harboured by the sawtoothed grain beetle Oryzaephilus surinamensis (Coleoptera, Silvanidae), through a combination of genomics approaches and bioassays. The Wolbachia strain wSur belongs to supergroup B that contains well-described reproductive manipulators of insects and encodes a pair of cytoplasmic incompatibility factor (cif) genes, as well as multiple homologues of the WO-mediated killing (wmk) gene. A phylogenetic comparison with wmk homologues of wMel of Drosophila melanogaster identified 18 wmk copies in wSur, including one that is closely related to the wMel male-killing homologue. However, further analysis of this particular wmk gene revealed an eight-nucleotide deletion leading to a stop-codon and subsequent reading frame shift midsequence, probably rendering it nonfunctional. Concordantly, utilizing a Wolbachia-deprived O. surinamensis population and controlled mating pairs of wSur-infected and noninfected partners, we found no experimental evidence for male-killing. However, a significant ~50% reduction of hatching rates in hybrid crosses of uninfected females with infected males indicates that wSur is causing cytoplasmic incompatibility. Thus, Wolbachia also represents an important determinant of host fitness in Coleoptera.}, } @article {pmid36200325, year = {2022}, author = {Ross, PA and Hoffmann, AA}, title = {Fitness costs of Wolbachia shift in locally-adapted Aedes aegypti mosquitoes.}, journal = {Environmental microbiology}, volume = {24}, number = {12}, pages = {5749-5759}, doi = {10.1111/1462-2920.16235}, pmid = {36200325}, issn = {1462-2920}, mesh = {Animals ; Female ; *Wolbachia/genetics ; *Aedes/genetics ; }, abstract = {Aedes aegypti mosquito eggs can remain quiescent for many months before hatching, allowing populations to persist through unfavourable conditions. A. aegypti infected with the Wolbachia strain wMel have been released in tropical and subtropical regions for dengue control. wMel reduces the viability of quiescent eggs, but this physiological cost might be expected to evolve in natural mosquito populations that frequently experience stressful conditions. We found that the cost of wMel infection differed consistently between mosquitoes collected from different locations and became weaker across laboratory generations, suggesting environment-specific adaptation of mosquitoes to the wMel infection. Reciprocal crossing experiments show that differences in the cost of wMel to quiescent egg viability were mainly due to mosquito genetic background and not Wolbachia origin. wMel-infected mosquitoes hatching from long-term quiescent eggs showed partial loss of cytoplasmic incompatibility and female infertility, highlighting additional costs of long-term quiescence. Our study provides the first evidence for a shift in Wolbachia phenotypic effects following deliberate field release and establishment and it highlights interactions between Wolbachia infections and mosquito genetic backgrounds. The unexpected changes in fitness costs observed here suggest potential tradeoffs with undescribed fitness benefits of the wMel infection.}, } @article {pmid36192576, year = {2022}, author = {Štarhová Serbina, L and Gajski, D and Malenovský, I and Corretto, E and Schuler, H and Dittmer, J}, title = {Wolbachia infection dynamics in a natural population of the pear psyllid Cacopsylla pyri (Hemiptera: Psylloidea) across its seasonal generations.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {16502}, pmid = {36192576}, issn = {2045-2322}, mesh = {Animals ; Female ; *Hemiptera/genetics ; Male ; Phylogeny ; *Pyrus ; Seasons ; *Wolbachia/genetics ; }, abstract = {Wolbachia is one of the most abundant intracellular symbionts of arthropods and has profound effects on host biology. Wolbachia transmission and host phenotypes often depend on its density within the host, which can be affected by multiple biotic and abiotic factors. However, very few studies measured Wolbachia density in natural host populations. Here, we describe Wolbachia in the pear psyllid Cacopsylla pyri from three populations in the Czech Republic. Using phylogenetic analyses based on wsp and multilocus sequence typing genes, we demonstrate that C. pyri harbours three new Wolbachia strains from supergroup B. A fourth Wolbachia strain from supergroup A was also detected in parasitised immatures of C. pyri, but likely came from a hymenopteran parasitoid. To obtain insights into natural Wolbachia infection dynamics, we quantified Wolbachia in psyllid individuals from the locality with the highest prevalence across an entire year, spanning several seasonal generations of the host. All tested females were infected and Wolbachia density remained stable across the entire period, suggesting a highly efficient vertical transmission and little influence from the environment and different host generations. In contrast, we observed a tendency towards reduced Wolbachia density in males which may suggest sex-related differences in Wolbachia-psyllid interactions.}, } @article {pmid36182681, year = {2022}, author = {Ooi, EE and Wilder-Smith, A}, title = {Externalities modulate the effectiveness of the Wolbachia release programme.}, journal = {The Lancet. Infectious diseases}, volume = {22}, number = {11}, pages = {1518-1519}, doi = {10.1016/S1473-3099(22)00497-2}, pmid = {36182681}, issn = {1474-4457}, mesh = {Animals ; Humans ; *Wolbachia ; *Aedes ; *Dengue ; }, } @article {pmid36182679, year = {2022}, author = {Ribeiro Dos Santos, G and Durovni, B and Saraceni, V and Souza Riback, TI and Pinto, SB and Anders, KL and Moreira, LA and Salje, H}, title = {Estimating the effect of the wMel release programme on the incidence of dengue and chikungunya in Rio de Janeiro, Brazil: a spatiotemporal modelling study.}, journal = {The Lancet. Infectious diseases}, volume = {22}, number = {11}, pages = {1587-1595}, pmid = {36182679}, issn = {1474-4457}, mesh = {Humans ; Animals ; *Chikungunya Fever/epidemiology/prevention & control ; *Dengue Virus ; Incidence ; Brazil/epidemiology ; *Wolbachia ; *Aedes ; *Dengue/epidemiology/prevention & control ; Mosquito Vectors ; }, abstract = {BACKGROUND: Introgression of genetic material from species of the insect bacteria Wolbachia into populations of Aedes aegypti mosquitoes has been shown in randomised and non-randomised trials to reduce the incidence of dengue; however, evidence for the real-world effectiveness of large-scale deployments of Wolbachia-infected mosquitoes for arboviral disease control in endemic settings is still scarce. A large Wolbachia (wMel strain) release programme was implemented in 2017 in Rio de Janeiro, Brazil. We aimed to assess the effect of this programme on the incidence of dengue and chikungunya in the city.

METHODS: 67 million wMel-infected mosquitoes were released across 28 489 locations over an area of 86·8 km[2] in Rio de Janeiro between Aug 29, 2017 and Dec 27, 2019. Following releases, mosquitoes were trapped and the presence of wMel was recorded. In this spatiotemporal modelling study, we assessed the effect of the release programme on the incidence of dengue and chikungunya. We used spatiotemporally explicit mathematical models applied to geocoded dengue cases (N=283 270) from 2010 to 2019 and chikungunya cases (N=57 705) from 2016 to 2019.

FINDINGS: On average, 32% of mosquitoes collected from the release zones between 1 month and 29 months after the initial release tested positive for wMel. Reduced wMel introgression occurred in locations and seasonal periods in which cases of dengue and chikungunya were historically high, with a decrease to 25% of mosquitoes testing positive for wMel during months in which disease incidence was at its highest. Despite incomplete introgression, we found that the releases were associated with a 38% (95% CI 32-44) reduction in the incidence of dengue and a 10% (4-16) reduction in the incidence of chikungunya.

INTERPRETATION: Stable establishment of wMel in the geographically diverse, urban setting of Rio de Janeiro seems to be more complicated than has been observed elsewhere. However, even intermediate levels of wMel seem to reduce the incidence of disease caused by two arboviruses. These findings will help to guide future release programmes.

FUNDING: Bill & Melinda Gates Foundation and the European Research Council.}, } @article {pmid36172295, year = {2022}, author = {Tiwary, A and Babu, R and Sen, R and Raychoudhury, R}, title = {Bacterial supergroup-specific "cost" of Wolbachia infections in Nasonia vitripennis.}, journal = {Ecology and evolution}, volume = {12}, number = {9}, pages = {e9219}, pmid = {36172295}, issn = {2045-7758}, abstract = {The maternally inherited endosymbiont, Wolbachia, is known to alter the reproductive biology of its arthropod hosts for its own benefit and can induce both positive and negative fitness effects in many hosts. Here, we describe the effects of the maintenance of two distinct Wolbachia infections, one each from supergroups A and B, on the parasitoid host Nasonia vitripennis. We compare the effect of Wolbachia infections on various traits between the uninfected, single A-infected, single B-infected, and double-infected lines with their cured versions. Contrary to some previous reports, our results suggest that there is a significant cost associated with the maintenance of Wolbachia infections where traits such as family size, fecundity, longevity, and rates of male copulation are compromised in Wolbachia-infected lines. The double Wolbachia infection has the most detrimental impact on the host as compared to single infections. Moreover, there is a supergroup-specific negative impact on these wasps as the supergroup B infection elicits the most pronounced negative effects. These negative effects can be attributed to a higher Wolbachia titer seen in the double and the single supergroup B infection lines when compared to supergroup A. Our findings raise important questions on the mechanism of survival and maintenance of these reproductive parasites in arthropod hosts.}, } @article {pmid36169529, year = {2022}, author = {Favoreto, AL and Carvalho, VR and Domingues, MM and Ribeiro, MF and Cavallini, G and Lawson, SA and Silva, WM and Zanuncio, JC and Wilcken, CF}, title = {Wolbachia pipientis: first detection in populations of Glycaspis brimblecombei (Hemiptera: Aphalaridae) and Psyllaephagus bliteus (Hymenoptera: Encyrtidae) in Brazil.}, journal = {Brazilian journal of biology = Revista brasleira de biologia}, volume = {82}, number = {}, pages = {e264475}, doi = {10.1590/1519-6984.264475}, pmid = {36169529}, issn = {1678-4375}, mesh = {Animals ; Brazil ; *Eucalyptus ; *Hemiptera ; Humans ; *Hymenoptera ; *Wolbachia ; }, abstract = {The sucking insect, Glycaspis brimblecombei Moore (Hemiptera: Aphalaridae), is originally from Australia and reduces the productivity of Eucalyptus crops. The parasitoid Psyllaephagus bliteus Riek (Hymenoptera: Encyrtidae) is the main agent used in the integrated management of G. brimblecombei. Endosymbionts, in insects, are important in the adaptation and protection of their hosts to the environment. The intracellular symbionts Wolbachia, induces reproductive changes such as cytoplasmic incompatibility, feminization, male death and parthenogenesis. The objective of this study was to report the first record of Wolbachia pipientis in populations of G. brimblecombei and of its parasitoid P. bliteus in the field in Brazil. Branches with adults of G. brimblecombei and P. bliteus were collected from eucalyptus trees in commercial farms in six Brazilian states and, after emergence, the insects obtained were frozen at -20 °C. Polymerase chain reaction (PCR) was performed to detect the Wolbachia endosymbiont. Wolbachia pipientis was identified in individuals of G. brimblecombei and its parasitoid P. bliteus from populations of the counties of Agudos and Mogi-Guaçu (São Paulo State), Itamarandiba (Minas Gerais State) and São Jerônimo da Serra (Paraná State) in Brazil.}, } @article {pmid36168763, year = {2022}, author = {Dudzic, JP and Curtis, CI and Gowen, BE and Perlman, SJ}, title = {A highly divergent Wolbachia with a tiny genome in an insect-parasitic tylenchid nematode.}, journal = {Proceedings. Biological sciences}, volume = {289}, number = {1983}, pages = {20221518}, pmid = {36168763}, issn = {1471-2954}, mesh = {Animals ; Heme ; Insecta ; *Nematoda/genetics ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia symbionts are the most successful host-associated microbes on the planet, infecting arthropods and nematodes. Their role in nematodes is particularly enigmatic, with filarial nematode species either 100% infected and dependent on symbionts for reproduction and development, or not at all infected. We have discovered a highly divergent strain of Wolbachia in an insect-parasitic tylenchid nematode, Howardula sp., in a nematode clade that has not previously been known to harbour Wolbachia. While this nematode is 100% infected with Wolbachia, we did not detect it in related species. We sequenced the Howardula symbiont (wHow) genome and found that it is highly reduced, comprising only 550 kilobase pairs of DNA, approximately 35% smaller than the smallest Wolbachia nematode symbiont genomes. The wHow genome is a subset of all other Wolbachia genomes and has not acquired any new genetic information. While it has lost many genes, including genes involved in cell wall synthesis and cell division, it has retained the entire haem biosynthesis pathway, suggesting that haem supplementation is critical. wHow provides key insights into our understanding of what are the lower limits of Wolbachia cells, as well as the role of Wolbachia symbionts in the biology and convergent evolution of diverse parasitic nematodes.}, } @article {pmid36165808, year = {2022}, author = {Hussain, M and Bradshaw, T and Lee, M and Asgari, S}, title = {The Involvement of Atlastin in Dengue Virus and Wolbachia Infection in Aedes aegypti and Its Regulation by aae-miR-989.}, journal = {Microbiology spectrum}, volume = {10}, number = {5}, pages = {e0225822}, pmid = {36165808}, issn = {2165-0497}, mesh = {Animals ; *Aedes/microbiology/virology ; *Dengue ; *Dengue Virus/genetics/metabolism ; Furin/metabolism ; *MicroRNAs/genetics/metabolism ; Virus Replication/physiology ; *Wolbachia/genetics ; }, abstract = {Endoplasmic reticulum (ER)-shaping atlastin proteins (ATLs) have been demonstrated to play a functional role during flavivirus replication in mammalian cells. For dengue virus (DENV), atlastin is required in the formation of the replication organelles and RNA replication, virion assembly, production of the infectious virus particles, and trafficking or directing the association of vesicle packets with furin. Here, we investigated the involvement of atlastin in DENV replication in the mosquito Aedes aegypti and explored the possibility of its manipulation by the endosymbiotic bacterium Wolbachia to interfere with DENV replication. Results showed the expression of Ae. aegypti atlastin gene (AaATL) was upregulated in DENV-infected Aag2 cells, and its silencing led to reduced DENV replication. Contrary to our assumption that AaATL could be downregulated by Wolbachia, we did not find evidence for that in Wolbachia-infected cell lines, but this was the case in mosquitoes. Further, silencing AaATL did not have any effect on Wolbachia density. Our results also suggest that aae-miR-989 miRNA negatively regulates AaATL. The oversupply of the miRNA mimic led to reduced DENV replication consistent with the positive role of AaATL in DENV replication. Overall, the results favor AaATL's involvement in DENV replication; however, there is no support that the protein is involved in Wolbachia-mediated DENV inhibition. In addition, the results contribute to discerning further possible overlapping functions of ATLs in mosquitoes and mammalian cells. IMPORTANCE Atlastin is a protein associated with the endoplasmic reticulum and has been shown to play a role in replication of flaviviruses in mammalian cells. This study aimed to investigate the role of mosquito Aedes aegypti atlastin (AaATL) in dengue virus replication and maintenance of Wolbachia, an endosymbiotic bacterium, in the mosquito. Our results suggest that AaATL facilitates dengue virus replication in mosquito cells, considering silencing the gene led to reductions in virus replication and virion production. Further, AaATL was found to be regulated by a mosquito microRNA, aae-miR-989. Despite an effect on dengue virus, AaATL silencing did not affect Wolbachia replication and maintenance in mosquito cells. The results shed light on the role of atlastins in mosquito-pathogen interactions and their overlapping roles in mosquito and mammalian cells.}, } @article {pmid36151871, year = {2023}, author = {Bing, XL and Xia, CB and Ye, QT and Gong, X and Cui, JR and Peng, CW and Hong, XY}, title = {Wolbachia manipulates reproduction of spider mites by influencing herbivore salivary proteins.}, journal = {Pest management science}, volume = {79}, number = {1}, pages = {315-323}, doi = {10.1002/ps.7201}, pmid = {36151871}, issn = {1526-4998}, support = {//Fundamental Research Funds for the Central Universities/ ; //National Natural Science Foundation of China/ ; //Natural Science Foundation of Jiangsu Province/ ; //startup grant from Nanjing Agricultural University/ ; }, mesh = {Animals ; *Tetranychidae ; *Wolbachia ; Proteomics ; Salivary Proteins and Peptides/genetics ; }, abstract = {BACKGROUND: The endosymbiont Wolbachia is known for manipulating host reproduction. Wolbachia also can affect host fitness by mediating interactions between plant and herbivores. However, it remains unclear whether saliva proteins are involved in this process.

RESULTS: We found that Wolbachia infection decreased the number of deposited eggs but increased the egg hatching rate in the spider mite Tetranychus urticae Koch (Acari: Tetranychidae), a cosmopolitan pest that infects >1000 species of plants. Transcriptomic and proteomic analyses revealed that Wolbachia-infected mites upregulated the gene expression levels of many T. urticae salivary proteins including a cluster of Tetranychidae-specific, functionally uncharacterized SHOT1s (secreted host-responsive proteins of Tetranychidae). The SHOT1 genes were expressed more in the feeding stages (nymphs and adults) of mites than in eggs and highly enriched in the proterosomas. RNA interference experiments showed that knockdown of SHOT1s significantly decreased Wolbachia density, increased the number of deposited eggs and decreased the egg hatching rate.

CONCLUSION: Together, these results indicate that SHOT1s are positively correlated with Wolbachia density and account for Wolbachia-mediated phenotypes. Our results provide new evidence that herbivore salivary proteins are related to Wolbachia-mediated manipulations of host performance on plants. © 2022 Society of Chemical Industry.}, } @article {pmid36149408, year = {2022}, author = {Warecki, B and Titen, SWA and Alam, MS and Vega, G and Lemseffer, N and Hug, K and Minden, JS and Sullivan, W}, title = {Wolbachia action in the sperm produces developmentally deferred chromosome segregation defects during the Drosophila mid-blastula transition.}, journal = {eLife}, volume = {11}, number = {}, pages = {}, pmid = {36149408}, issn = {2050-084X}, support = {R35 GM139595/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Blastula ; Chromatin ; Chromosome Segregation ; Cytoplasm ; Drosophila/genetics ; Female ; In Situ Hybridization, Fluorescence ; Male ; Semen ; Spermatozoa ; *Wolbachia/genetics ; }, abstract = {Wolbachia, a vertically transmitted endosymbiont infecting many insects, spreads rapidly through uninfected populations by a mechanism known as cytoplasmic incompatibility (CI). In CI, a paternally delivered modification of the sperm leads to chromatin defects and lethality during and after the first mitosis of embryonic development in multiple species. However, whether CI-induced defects in later stage embryos are a consequence of the first division errors or caused by independent defects remains unresolved. To address this question, we focused on ~1/3 of embryos from CI crosses in Drosophila simulans that develop apparently normally through the first and subsequent pre-blastoderm divisions before exhibiting mitotic errors during the mid-blastula transition and gastrulation. We performed single embryo PCR and whole genome sequencing to find a large percentage of these developed CI-derived embryos bypass the first division defect. Using fluorescence in situ hybridization, we find increased chromosome segregation errors in gastrulating CI-derived embryos that had avoided the first division defect. Thus, Wolbachia action in the sperm induces developmentally deferred defects that are not a consequence of the first division errors. Like the immediate defect, the delayed defect is rescued through crosses to infected females. These studies inform current models on the molecular and cellular basis of CI.}, } @article {pmid36147860, year = {2022}, author = {Guo, L and Tang, C and Gao, C and Li, Z and Cheng, Y and Chen, J and Wang, T and Xu, J}, title = {Bacterial and fungal communities within and among geographic samples of the hemp pest Psylliodes attenuata from China.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {964735}, pmid = {36147860}, issn = {1664-302X}, abstract = {The hemp flea beetle Psylliodes attenuata (Coleoptera: Chrysomelidae: Psylliodes) is a common pest of Cannabis sativa, including cultivars of both medicinal marijuana and industrial hemp. Both the larval and adult stages of this beetle can cause significant damages to C. sativa, resulting in substantial crop losses. At present, little is known about the bacterial and fungal community diversity among populations of this pest insect. In the present study, we obtained P. attenuata samples from nine field sites representing broad industrial hemp productions in China and analyzed their microbial communities using DNA metabarcoding. Bacterial sequences of all the samples were assigned to 3728 OTUs, which belonged to 45 phyla, 1058 genera and 1960 known species. The most common genera were Rickettsia, Wolbachia, and Candidatus_Brownia. Fungal sequences of all the samples were assigned to 910 OTUs, which belonged to 9 phyla, 308 genera and 464 known species. The most common fungal genera were Cladosporium, Cutaneotrichosporon, and Aspergillus. Principal coordinate analysis revealed a significant difference in the bacterial and fungal community structure among the nine P. attenuata populations. Understanding the microbial symbionts may provide clues to help develop potential biocontrol techniques against this pest.}, } @article {pmid36147838, year = {2022}, author = {Liu, YH and Ma, YM and Tian, HO and Yang, B and Han, WX and Zhao, WH and Chai, HL and Zhang, ZS and Wang, LF and Chen, L and Xing, Y and Ding, YL and Zhao, L}, title = {First determination of DNA virus and some additional bacteria from Melophagus ovinus (sheep ked) in Tibet, China.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {988136}, pmid = {36147838}, issn = {1664-302X}, abstract = {Melophagus ovinus (sheep ked) is one of the common ectoparasites in sheep. In addition to causing direct damage to the host through biting and sucking blood, sheep ked is a potential vector of helminths, protozoa, bacteria, and viruses. Sheep M. ovinus samples from three regions in Tibet were selected for DNA extraction. The 16S rDNA V3-V4 hypervariable region was amplified, after genomic DNA fragmentation, Illumina Hiseq libraries were constructed. The 16S rRNA sequencing and viral metagenomics sequencing were separately conducted on the Illumina Novaseq 6000 platform and molecular biology software and platforms were employed to analyze the sequencing data. Illumina PE250 sequencing results demonstrated that the dominant bacteria phylum in M. ovinus from Tibet, China was Proteobacteria, where 29 bacteria genera were annotated. The dominant bacterial genera were Bartonella, Wolbachia, and Arsenophonus; Bartonella chomelii, Wolbachia spp., and Arsenophonus spp. were the dominant bacterial species in M. ovinus from Tibet, China. We also detected Kluyvera intermedia, Corynebacterium maris DSM 45190, Planomicrobium okeanokoites, and Rhodococcus erythropolis, of which the relative abundance of Kluyvera intermedia was high. Illumina Hiseq sequencing results demonstrated that 4 virus orders were detected in M. ovinus from Tibet, China, and 3 samples were annotated into 29 families, 30 families, and 28 families of viruses, respectively. Virus families related to vertebrates and insects mainly included Mimiviridae, Marseilleviridae, Poxviridae, Ascoviridae, Iridoviridae, Baculoviridae, Hytrosaviridae, Nudiviridae, Polydnaviridae, Adomaviridae, Asfarviridae, Hepeviridae, Herpesviridae, and Retroviridae; at the species level, the relative abundance of Tupanvirus_soda_lake, Klosneuvirus_KNV1, and Indivirus_ILV1 was higher. African swine fever virus and many poxviruses from the family Poxviridae were detected, albeit their relative abundance was low. The dominant bacterial phylum of M. ovinus from Tibet, China was Proteobacteria, and the dominant bacterial genera were Bartonella, Wolbachia, and Arsenophonus, where 23 out of 29 annotated bacteria genera were first reported in M. ovinus. Kluyvera intermedia, Corynebacterium maris DSM 45190, Planomicrobium okeanokoites, and Rhodococcus erythropolis were detected for the first time. All DNA viruses detected in this study have been reported in M. ovinus for the first time.}, } @article {pmid36138209, year = {2023}, author = {Liu, Q and Zhang, H and Huang, X}, title = {Strong Linkage Between Symbiotic Bacterial Community and Host Age and Morph in a Hemipteran Social Insect.}, journal = {Microbial ecology}, volume = {86}, number = {2}, pages = {1213-1225}, pmid = {36138209}, issn = {1432-184X}, support = {31970446//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Female ; Biological Evolution ; *Aphids/microbiology ; Insecta ; Symbiosis/physiology ; *Buchnera/genetics ; }, abstract = {The relationships between symbionts and insects are complex, and symbionts usually have diverse ecological and evolutionary effects on their hosts. The phloem sap-sucking aphids are good models to study the interactions between insects and symbiotic microorganisms. Although aphids usually exhibit remarkable life cycle complexity, most previous studies on symbiotic diversity sampled only apterous viviparous adult females or very few morphs. In this study, high-throughput 16S rDNA amplicon sequencing was used to assess the symbiotic bacterial communities of eleven morphs or developmental stages of the social aphid Pseudoregma bambucicola. We found there were significant differences in bacterial composition in response to different morphs and developmental stages, and for the first time, we revealed male aphids hosted very different symbiotic composition featured with low abundance of dominant symbionts but high diversity of total symbionts. The relative abundance of Pectobacterium showed relatively stable across different types of samples, while that of Wolbachia fluctuated greatly, indicating the former may have a consistent function in this species and the latter may provide specific function for certain morphs or developmental stages. Our study presents new evidence of complexity of symbiotic associations and indicates strong linkage between symbiotic bacterial community and host age and morph.}, } @article {pmid36135489, year = {2022}, author = {Andrianto, E and Kasai, A}, title = {Wolbachia in Black Spiny Whiteflies and Their New Parasitoid Wasp in Japan: Evidence of the Distinct Infection Status on Aleurocanthus camelliae Cryptic Species Complex.}, journal = {Insects}, volume = {13}, number = {9}, pages = {}, pmid = {36135489}, issn = {2075-4450}, abstract = {Wolbachia, an alphaproteobacterial reproductive parasite, can cause profound mitochondrial divergence in insects, which might eventually be a part of cryptic speciation. Aleurocanthus camelliae is a cryptic species complex consisting of several morphospecies and/or haplotypes that are genetically different but morphologically indistinctive. However, little is known about the Wolbachia infection status in these tea and Citrus pests. Thus, this study aimed to profile the diversity and phenotypic characteristics of Wolbachia natural infections in the A. camelliae cryptic species complex. A monophyletic strain of Wolbachia that infected the A. camelliae cryptic species complex (wAlec) with different patterns was discovered. Whiteflies that are morphologically identical to Aleurocanthus spiniferus (Aleurocanthus cf. A. spiniferus in Eurya japonica and A. spiniferus in Citrus) were grouped into uninfected populations, whereas the fixed infection was detected in A. camelliae B1 from Theaceae. The rapid evolution of wAlec was also found to occur through a high recombination event, which produced subgroups A and B in wAlec. It may also be associated with the non-cytoplasmic incompatibility (CI) phenotype of wAlec due to undetectable CI-related genes from phage WO (WOAlec). The current discovery of a novel cryptic species of A. camelliae led to a discussion about the oscillation hypothesis, which may provide insights on cryptic speciation, particularly on how specialization and host expansion have been recorded among these species. This study also identified a parasitoid wasp belonging to the genus Eretmocerus in A. camelliae, for the first time in Japan.}, } @article {pmid36125236, year = {2023}, author = {Brinker, P and Chen, F and Chehida, YB and Beukeboom, LW and Fontaine, MC and Salles, JF}, title = {Microbiome composition is shaped by geography and population structure in the parasitic wasp Asobara japonica, but not in the presence of the endosymbiont Wolbachia.}, journal = {Molecular ecology}, volume = {32}, number = {23}, pages = {6644-6658}, doi = {10.1111/mec.16699}, pmid = {36125236}, issn = {1365-294X}, mesh = {Animals ; *Wasps/genetics/microbiology ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; *Microbiota/genetics ; Bacteria/genetics ; Geography ; }, abstract = {The microbial community composition is crucial for diverse life-history traits in many organisms. However, we still lack a sufficient understanding of how the host microbiome is acquired and maintained, a pressing issue in times of global environmental change. Here we investigated to what extent host genotype, environmental conditions, and the endosymbiont Wolbachia influence the bacterial communities in the parasitic wasp Asobara japonica. We sampled multiple wasp populations across 10 locations in their natural distribution range in Japan and sequenced the host genome (whole genome sequencing) and microbiome (16S rRNA gene). We compared the host population structure and bacterial community composition of wasps that reproduce sexually and are uninfected with Wolbachia with wasps that reproduce asexually and carry Wolbachia. The bacterial communities in asexual wasps were highly similar due to a strong effect of Wolbachia rather than host genomic structure. In contrast, in sexual wasps, bacterial communities appear primarily shaped by a combination of population structure and environmental conditions. Our research highlights that multiple factors shape the bacterial communities of an organism and that the presence of a single endosymbiont can strongly alter their compositions. This information is crucial to understanding how organisms and their associated microbiome will react in the face of environmental change.}, } @article {pmid36124585, year = {2022}, author = {Chinnathambi, R and Rihan, FA}, title = {Analysis and control of Aedes Aegypti mosquitoes using sterile-insect techniques with Wolbachia.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {19}, number = {11}, pages = {11154-11171}, doi = {10.3934/mbe.2022520}, pmid = {36124585}, issn = {1551-0018}, mesh = {*Aedes ; Animals ; Humans ; *Infertility ; Male ; Mosquito Control/methods ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Combining Sterile and Incompatible Insect techniques can significantly reduce mosquito populations and prevent the transmission of diseases between insects and humans. This paper describes impulsive differential equations for the control of a mosquito with Wolbachia. Several interesting conditions are created when sterile male mosquitoes are released impulsively, ensuring both open- and closed-loop control. To determine the wild mosquito population size in real-time, we propose an open-loop control system, which uses impulsive and constant releases of sterile male mosquitoes. A closed-loop control scheme is also being investigated, which specifies the release of sterile mosquitoes according to the size of the wild mosquito population. To eliminate or reduce a mosquito population below a certain threshold, the Sterile insect technique involves mass releases of sterile insects. Numerical simulations verify the theoretical results.}, } @article {pmid36121852, year = {2022}, author = {Martinez, J and Ant, TH and Murdochy, SM and Tong, L and da Silva Filipe, A and Sinkins, SP}, title = {Genome sequencing and comparative analysis of Wolbachia strain wAlbA reveals Wolbachia-associated plasmids are common.}, journal = {PLoS genetics}, volume = {18}, number = {9}, pages = {e1010406}, pmid = {36121852}, issn = {1553-7404}, support = {MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Aedes/genetics ; Animals ; Plasmids/genetics ; Prophages/genetics ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are widespread maternally-transmitted bacteria of arthropods that often spread by manipulating their host's reproduction through cytoplasmic incompatibility (CI). Their invasive potential is currently being harnessed in field trials aiming to control mosquito-borne diseases. Wolbachia genomes commonly harbour prophage regions encoding the cif genes which confer their ability to induce CI. Recently, a plasmid-like element was discovered in wPip, a Wolbachia strain infecting Culex mosquitoes; however, it is unclear how common such extra-chromosomal elements are in Wolbachia. Here we sequenced the complete genome of wAlbA, a strain of the symbiont found in Aedes albopictus, after eliminating the co-infecting and higher density wAlbB strain that previously made sequencing of wAlbA challenging. We show that wAlbA is associated with two new plasmids and identified additional Wolbachia plasmids and related chromosomal islands in over 20% of publicly available Wolbachia genome datasets. These plasmids encode a variety of accessory genes, including several phage-like DNA packaging genes as well as genes potentially contributing to host-symbiont interactions. In particular, we recovered divergent homologues of the cif genes in both Wolbachia- and Rickettsia-associated plasmids. Our results indicate that plasmids are common in Wolbachia and raise fundamental questions around their role in symbiosis. In addition, our comparative analysis provides useful information for the future development of genetic tools to manipulate and study Wolbachia symbionts.}, } @article {pmid36117539, year = {2022}, author = {Power, NR and Rugman-Jones, PF and Stouthamer, R and Ganjisaffar, F and Perring, TM}, title = {High temperature mortality of Wolbachia impacts the sex ratio of the parasitoid Ooencyrtus mirus (Hymenoptera: Encyrtidae).}, journal = {PeerJ}, volume = {10}, number = {}, pages = {e13912}, pmid = {36117539}, issn = {2167-8359}, mesh = {Animals ; Male ; Female ; Temperature ; *Wolbachia/genetics ; Sex Ratio ; *Wasps/microbiology ; Reproduction ; *Heteroptera ; }, abstract = {BACKGROUND: Wolbachia bacteria are estimated to occur in more than half of all insect species. In Hymenoptera, Wolbachia often manipulates its host's reproduction to its own advantage. Wolbachia is likely the reason that males are rare in the uniparental Ooencyrtus mirus Triapitsyn & Power (Hymenoptera: Encyrtidae). The likelihood of producing male offspring can be increased by giving mothers a continuous supply of Bagrada hilaris (Burmeister) (Heteroptera: Pentatomidae) host eggs to parasitize for 2-3 weeks, by feeding the parents antibiotics, or by rearing parent wasps at high temperatures; all variables that have been shown to correlate with depleting Wolbachia titers in other organisms. The purpose of the current study was to determine whether thelytoky in O. mirus is due to Wolbachia, and if so, at what time in development the sex change occurs. We also wished to determine if Wolbachia removal results in the production of intersexes, as in some other hymenopterans. Finally, mating behavior was observed to see if and where it breaks down as a result of the species becoming thelytokous.

METHODS: Females were collected from parental lines of O. mirus reared at 26, 30, 31, 32, 33, 34, and 36 °C. The offspring of these females were reared at 26 °C, and their sex-ratio was determined. In a subsequent experiment, the parental generation was switched between 26 °C and 36 °C during development to narrow down the critical period at which changes occurred that subsequently affected the sex-ratio of their offspring.

RESULTS: The sex ratio was male biased in the offspring of O. mirus parents reared at 34 °C and 36 °C (high temperatures), even if the offspring themselves were reared at 26 °C. The constant temperature at which the percentage of males started to increase after two generations was 31 °C (10% males), rising to 39% males at 33 °C, and 100% males at 34 °C and 36 °C. Lasting more than 2 days, the critical period for the change toward a male biased sex ratio was during the second half of the parent's development. Molecular diagnostic assays confirmed that O. mirus females contain Wolbachia and males do not. Examination of preserved males and male-female pairs under a dissecting microscope showed no signs of intersex characters. Observation of the mating behavior of live O. mirus showed that males initiate courtship by drumming their antennae on a female's antennae, but after a few seconds, the females typically turn and walk away. However, a few instances of possible copulation were noted.

CONCLUSIONS: As hypothesized, the results indicated that thelytoky in O. mirus is likely mediated by Wolbachia bacteria. To maximize the population growth rate without generating males, the best temperature for mass rearing this species is 30 °C.}, } @article {pmid36114796, year = {2022}, author = {Li, J and He, P and He, P and Li, Y and Wu, Y and Lu, Z and Li, X and Yang, Y and Wang, Y and Guo, J and Munir, S and He, Y}, title = {Potential of citrus endophyte Bacillus subtilis L1-21 in the control of Candidatus Liberibacter asiaticus in Asian citrus psyllid, Diaphorina citri.}, journal = {Pest management science}, volume = {78}, number = {12}, pages = {5164-5171}, doi = {10.1002/ps.7134}, pmid = {36114796}, issn = {1526-4998}, support = {//Central Government Fund for Local Science and Technology Development/ ; //China Postdoctoral Science Foundation/ ; //National Natural Science Foundation of China/ ; //Yunnan First Level Research Fund for Post-doctorate Researchers/ ; //Yunnan Oriental Training Program Funding for Post-doctorate researchers/ ; }, mesh = {Animals ; *Hemiptera/microbiology ; *Citrus/microbiology ; *Rhizobiaceae ; Endophytes ; Bacillus subtilis ; Liberibacter ; Insect Vectors/microbiology ; Plant Diseases/microbiology ; }, abstract = {BACKGROUND: Asian citrus psyllid (ACP), also known as Diaphorina citri, is the natural vector of Candidatus Liberibacter asiaticus (CLas), which is responsible for Huanglongbing (HLB), a devastating citrus disease. Previously, the pathogen was successfully excluded from diseased citrus plants by using the indigenous endophyte Bacillus subtilis L1-21. However, the pathogen elimination and colonization potential of B. subtilis L1-21 in the carrier vector ACP, as well as the recruitment of native microbial communities of psyllid in the presence of endophytes, are still unknown.

RESULTS: Initially, we suggested that endophyte L1-21 reduced the CLas copies in ACP from 6.58 × 10[6] to 5.04 × 10[4] per insect after 48 h, however, the pathogen copies remained stable in the negative control. The endophyte was stable for 48 h after application. Among the bacterial genera those highlighted in ACP were Candidatus Liberibacter, Pseudomonas, Candidatus Profftella, Methylobacterium-Methylorubrum, Pantoea, Curtobacterium, Wolbachia, Actinomycetospora, and Bacillus. Interestingly, B. subtilis L1-21 easily colonizes the midgut of ACP but cannot be detected in eggs. When ACP with endophyte L1-21 was allowed to feed on new citrus leaves, the highest colonization was observed. We also found that psyllids carrying endophyte L1-21 after feeding on citrus leaves reduced the CLas copies in leaves on the 0, 3rd and 5th day from 8.18 × 10,[4] 2.6 × 10,[3] and 0 pathogen copies/g fresh midvein, respectively.

CONCLUSIONS: We propose that B. subtilis L1-21 is a native endophyte in citrus and psyllid, which efficiently reduces the CLas pathogen in both citrus and psyllids, provides a more protective effect by increasing the number of cultivable endophytes, and successfully colonizes the midgut of ACP. © 2022 Society of Chemical Industry.}, } @article {pmid36112293, year = {2022}, author = {Liu, Y and Yu, J and Li, J}, title = {A Mosquito Population Suppression Model by Releasing Wolbachia-Infected Males.}, journal = {Bulletin of mathematical biology}, volume = {84}, number = {11}, pages = {121}, pmid = {36112293}, issn = {1522-9602}, mesh = {*Aedes ; Animals ; Male ; Mathematical Concepts ; Models, Biological ; *Wolbachia ; }, abstract = {Due to the role of cytoplasmic incompatibility (CI), releasing Wolbachia-infected male mosquitoes into the wild becomes a very promising strategy to suppress the wild mosquito population. When developing a mosquito suppression strategy, our main concerns are how often, and in what amount, should Wolbachia-infected mosquitoes be released under different CI intensity conditions, so that the suppression is most effective and cost efficient. In this paper, we propose a mosquito population suppression model that incorporates suppression and self-recovery under different CI intensity conditions. We adopt the new modeling idea that only sexually active Wolbachia-infected male mosquitoes are considered in the model and assume the releases of Wolbachia-infected male mosquitoes are impulsive and periodic with period T. We particularly study the case where the release period is greater than the sexual lifespan of the Wolbachia-infected male mosquitoes. We define the CI intensity threshold, mosquito release thresholds, and the release period threshold to characterize the model dynamics. The global and local asymptotic stability of the origin and the existence and stability of T-periodic solutions are investigated. Our findings provide useful guidance in designing practical release strategies to control wild mosquitoes.}, } @article {pmid36110209, year = {2022}, author = {Zhang, HD and Gao, J and Xing, D and Guo, XX and Li, CX and Dong, YD and Zheng, Z and Ma, Z and Wu, ZM and Zhu, XJ and Zhao, MH and Liu, QM and Yan, T and Chu, HL and Zhao, TY}, title = {Fine-scale genetic structure and wolbachia infection of aedes albopictus (Diptera: Culicidae) in Nanjing city, China.}, journal = {Frontiers in genetics}, volume = {13}, number = {}, pages = {827655}, pmid = {36110209}, issn = {1664-8021}, abstract = {Background: Aedes albopictus is an indigenous primary vector of dengue and Zika viruses in China. Wolbachia is a gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Compared with research on the dispersion of Ae. albopictus at the macrospatial level (mainly at the country or continent level), little is known about its variation and Wolbachia infection at the microspatial level, which is essential for its management. Meanwhile, no local cases of dengue fever have been recorded in the history of Nanjing, which implies that few adulticides have been applied in the city. Thus, the present study examines how the Ae. albopictus population varies and the Wolbachia infection status of each population among microspatial regions of Nanjing City. Methods: The genetic structure of 17 Aedes albopictus populations collected from urban, urban fringe, and rural regions of Nanjing City was investigated based on 9 microsatellite loci and the mitochondrial coxI gene. The Wolbachia infection status of each population was also assessed with Wolbachia A- and Wolbachia B-specific primers. Results: Nine out of 58 tested pairs of microsatellite markers were highly polymorphic, with a mean PIC value of 0.560, and these markers were therefore chosen for microsatellite genotyping analysis. The Na value of each Ae. albopictus population was very high, and the urban area populations (7.353 ± 4.975) showed a lower mean value than the urban fringe region populations (7.866 ± 5.010). A total of 19 coxI haplotypes were observed among 329 Ae. albopictus individuals via haplotype genotyping, with the highest diversity observed among the urban fringe Ae. albopictus populations (Hd = 0.456) and the lowest among the urban populations (Hd = 0.277). Each Ae. albopictus population showed significant departure from HWE, and significant population expansion was observed in only three populations from the urban (ZSL), urban fringe (HAJY), and rural areas (HSZY) (p < 0.05). Combined with DAPC analysis, all the Ae. albopictus populations were adequately allocated to two clades with significant genetic differences according to population structure analysis, and the best K value was equal to two. AMOVA results showed that most (96.18%) of the genetic variation detected in Ae. albopictus occurred within individuals (FIT = 0.22238, p < 0.0001), while no significant positive correlation was observed via isolation by distance (IBD) analysis (R [2] = 0.03262, p = 0.584). The TCS network of all haplotypes showed that haplotype 1 (H1) and haplotype 4 (H4) were the most frequent haplotypes among all populations, and the haplotype frequency significantly increased from urban regions (36.84%) to rural regions (68.42%). Frequent migration was observed among Ae. albopictus populations from rural to urban regions via the urban fringe region, with four direct migration routes between rural and urban regions. Furthermore, Wolbachia genotyping results showed that most of the individuals of each population were coinfected with Wolbachia A and Wolbachia B. The independent infection rate of Wolbachia A was slightly higher than that of Wolbachia B, and no significant differences were observed among different regions. Conclusion: In the microspatial environment of Nanjing City, the urban fringe region is an important region for the dispersion of Ae. albopictus populations between rural and urban areas, and Wolbachia A and Wolbachia B coinfection is the most common Wolbachia infection status in all Ae. albopictus populations among different regions.}, } @article {pmid36085160, year = {2022}, author = {Bishop, C and Hussain, M and Hugo, LE and Asgari, S}, title = {Analysis of Aedes aegypti microRNAs in response to Wolbachia wAlbB infection and their potential role in mosquito longevity.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {15245}, pmid = {36085160}, issn = {2045-2322}, mesh = {*Aedes/genetics ; Animals ; Anti-Bacterial Agents ; Drosophila ; Longevity/genetics ; *MicroRNAs/genetics ; Mosquito Vectors ; Tetracycline ; *Wolbachia ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {The mosquito Aedes aegypti is the primary vector of a range of medically important viruses including dengue, Zika, West Nile, yellow fever, and chikungunya viruses. The endosymbiotic bacterium Wolbachia pipientis wAlbB strain is a promising biocontrol agent for blocking viral transmission by Ae. aegypti. To predict the long-term efficacy of field applications, a thorough understanding of the interactions between symbiont, host, and pathogen is required. Wolbachia influences host physiology in a variety of ways including reproduction, immunity, metabolism, and longevity. MicroRNAs (miRNAs) are highly conserved small non-coding RNAs that regulate gene expression in eukaryotes and viruses. Several miRNAs are known to regulate biological processes in Drosophila and mosquitoes, including facilitating Wolbachia maintenance. We generated the first chromosomal map of Ae. aegypti miRNAs, and compared miRNA expression profiles between a wAlbB-transinfected Ae. aegypti mosquito line and a tetracycline cleared derivative, using deep small RNA-sequencing. We found limited modulation of miRNAs in response to wAlbB infection. Several miRNAs were modulated in response to age, some of which showed greater upregulation in wAlbB-infected mosquitoes than in tetracycline cleared ones. By selectively inhibiting some differentially expressed miRNAs, we identified miR-2946-3p and miR-317-3p as effecting mosquito longevity in Wolbachia-infected mosquitoes.}, } @article {pmid36042402, year = {2022}, author = {Twort, VG and Blande, D and Duplouy, A}, title = {One's trash is someone else's treasure: sequence read archives from Lepidoptera genomes provide material for genome reconstruction of their endosymbionts.}, journal = {BMC microbiology}, volume = {22}, number = {1}, pages = {209}, pmid = {36042402}, issn = {1471-2180}, mesh = {Animals ; *Lepidoptera ; Phylogeny ; *Spiroplasma/genetics ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Maternally inherited bacterial symbionts are extremely widespread in insects. They owe their success to their ability to promote their own transmission through various manipulations of their hosts' life-histories. Many symbionts however very often go undetected. Consequently, we have only a restricted idea of the true symbiont diversity in insects, which may hinder our understanding of even bigger questions in the field such as the evolution or establishment of symbiosis.

RESULTS: In this study, we screened publicly available Lepidoptera genomic material for two of the most common insect endosymbionts, namely Wolbachia and Spiroplasma, in 1904 entries, encompassing 106 distinct species. We compared the performance of two screening software, Kraken2 and MetaPhlAn2, to identify the bacterial infections and using a baiting approach we reconstruct endosymbiont genome assemblies. Of the 106 species screened, 20 (19%) and nine (8.5%) were found to be infected with either Wolbachia or Spiroplasma, respectively. Construction of partial symbiotic genomes and phylogenetic analyses suggested the Wolbachia strains from the supergroup B were the most prevalent type of symbionts, while Spiroplasma infections were scarce in the Lepidoptera species screened here.

CONCLUSIONS: Our results indicate that many of the host-symbiont associations remain largely unexplored, with the majority of associations we identify never being recorded before. This highlights the usefulness of public databases to explore the hidden diversity of symbiotic entities, allowing the development of hypotheses regarding host-symbiont associations. The ever-expanding genomic databases provide a diverse databank from which one can characterize and explore the true diversity of symbiotic entities.}, } @article {pmid36040904, year = {2022}, author = {Conjard, S and Meyer, DF and Aprelon, R and Pagès, N and Gros, O}, title = {Evidence of new strains of Wolbachia symbiont colonising semiaquatic bugs (Hemiptera: Gerroidea) in mangrove environment of the Lesser Antilles.}, journal = {PloS one}, volume = {17}, number = {8}, pages = {e0273668}, pmid = {36040904}, issn = {1932-6203}, mesh = {Animals ; *Arthropods/microbiology ; DNA, Bacterial ; *Heteroptera ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Wolbachia Hertig, 1936 is an intracellular bacterial symbiont colonizing many arthropods. Of the studies done on the bacteria present in the superfamily Gerroidea Leach, 1815, no report of Wolbachia infection had yet been made. Thus, we checked the presence of Wolbachia in six Gerroidea species which colonize tropical aquatic environments by PCR using wsp primer set before sequencing and phylogenetic analyses. Insects were collected in the marine fringe of mangroves, in river estuaries, in swampy mangroves, and in ponds from Guadeloupe islands (Caribbean). Two new strains of Wolbachia were detected in these Gerroidea. They were named wLfran and wRmang. The wsp sequences suggest that the strains belong to the already described E supergroup or similar. wLfran is present in Limnogonus franciscanus Stål, 1859 and Rheumatobates trinitatis (China, 1943) while wRmang appears to be present exclusively in R. mangrovensis (China, 1943). Three other species were analysed, but did not appear to be infected: Brachymetra albinerva (Amyot & Serville, 1843), Halobates micans Eschscheltz, 1822, and Microvelia pulchella Westwood, 1834. The results presented here highlight for the first time the presence of new intracellular Wolbachia strains in Gerroidea colonising tropical aquatic environments like mangrove habitats from inlands to sea shore.}, } @article {pmid36037217, year = {2022}, author = {Formisano, G and Iodice, L and Cascone, P and Sacco, A and Quarto, R and Cavalieri, V and Bosco, D and Guerrieri, E and Giorgini, M}, title = {Wolbachia infection and genetic diversity of Italian populations of Philaenus spumarius, the main vector of Xylella fastidiosa in Europe.}, journal = {PloS one}, volume = {17}, number = {8}, pages = {e0272028}, pmid = {36037217}, issn = {1932-6203}, mesh = {Animals ; Europe ; Genetic Variation ; *Hemiptera/genetics/microbiology ; Insect Vectors/microbiology ; Italy ; Multilocus Sequence Typing ; Plant Diseases/microbiology ; *Wolbachia/genetics ; *Xylella/genetics ; }, abstract = {Philaenus spumarius is a cosmopolitan species that has become a major threat to European agriculture being recognized as the main vector of the introduced plant pathogen Xylella fastidiosa, the agent of the "olive quick decline syndrome", a disease which is devastating olive orchards in southern Italy. Wolbachia are bacterial symbionts of many insects, frequently as reproductive parasites, sometime by establishing mutualistic relationships, able to spread within host populations. Philaenus spumarius harbors Wolbachia, but the role played by this symbiont is unknown and data on the infection prevalence within host populations are limited. Here, the Wolbachia infection rate was analyzed in relation to the geographic distribution and the genetic diversity of the Italian populations of P. spumarius. Analysis of the COI gene sequences revealed a geographically structured distribution of the three main mitochondrial lineages of P. spumarius. Wolbachia was detected in half of the populations sampled in northern Italy where most individuals belonged to the western-Mediterranean lineage. All populations sampled in southern and central Italy, where the individuals of the eastern-Mediterranean lineage were largely prevalent, were uninfected. Individuals of the north-eastern lineage were found only in populations from the Alps in the northernmost part of Italy, at high altitudes. In this area, Wolbachia infection reached the highest prevalence, with no difference between north-eastern and western-Mediterranean lineage. Analysis of molecular diversity of COI sequences suggested no significant effect of Wolbachia on population genetics of P. spumarius. Using the MLST approach, six new Wolbachia sequence types were identified. Using FISH, Wolbachia were observed within the host's reproductive tissues and salivary glands. Results obtained led us to discuss the role of Wolbachia in P. spumarius, the factors influencing the geographic distribution of the infection, and the exploitation of Wolbachia for the control of the vector insect to reduce the spread of X. fastidiosa.}, } @article {pmid36012723, year = {2022}, author = {Zong, Q and Mao, B and Zhang, HB and Wang, B and Yu, WJ and Wang, ZW and Wang, YF}, title = {Comparative Ubiquitome Analysis Reveals Deubiquitinating Effects Induced by Wolbachia Infection in Drosophila melanogaster.}, journal = {International journal of molecular sciences}, volume = {23}, number = {16}, pages = {}, pmid = {36012723}, issn = {1422-0067}, support = {31872288//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Cytoplasm/metabolism ; *Drosophila melanogaster/genetics ; Female ; Male ; Proteasome Endopeptidase Complex/metabolism ; Semen ; Testis/metabolism ; *Wolbachia ; }, abstract = {The endosymbiotic Wolbachia bacteria frequently cause cytoplasmic incompatibility (CI) in their insect hosts, where Wolbachia-infected males cross with uninfected females, leading to no or fewer progenies, indicating a paternal modification by Wolbachia. Recent studies have identified a Wolbachia protein, CidB, containing a DUB (deubiquitylating enzyme) domain, which can be loaded into host sperm nuclei and involved in CI, though the DUB activity is not necessary for CI in Drosophila melanogaster. To investigate whether and how Wolbachia affect protein ubiquitination in testes of male hosts and are thus involved in male fertility, we compared the protein and ubiquitinated protein expressions in D. melanogaster testes with and without Wolbachia. A total of 643 differentially expressed proteins (DEPs) and 309 differentially expressed ubiquitinated proteins (DEUPs) were identified to have at least a 1.5-fold change with a p-value of <0.05. Many DEPs were enriched in metabolic pathway, ribosome, RNA transport, and post-translational protein modification pathways. Many DEUPs were involved in metabolism, ribosome, and proteasome pathways. Notably, 98.1% DEUPs were downregulated in the presence of Wolbachia. Four genes coding for DEUPs in ubiquitin proteasome pathways were knocked down, respectively, in Wolbachia-free fly testes. Among them, Rpn6 and Rpn7 knockdown caused male sterility, with no mature sperm in seminal vesicles. These results reveal deubiquitylating effects induced by Wolbachia infection, suggesting that Wolbachia can widely deubiquitinate proteins that have crucial functions in male fertility of their hosts, but are not involved in CI. Our data provide new insights into the regulatory mechanisms of endosymbiont/host interactions and male fertility.}, } @article {pmid36009794, year = {2022}, author = {Suo, P and Wang, K and Yu, H and Fu, X and An, L and Bhowmick, B and Zhang, J and Han, Q}, title = {Seasonal Variation of Midgut Bacterial Diversity in Culexquinquefasciatus Populations in Haikou City, Hainan Province, China.}, journal = {Biology}, volume = {11}, number = {8}, pages = {}, pmid = {36009794}, issn = {2079-7737}, support = {ZDYF2021xx//Key Science and Technology Program of Hainan Province (ZDYF2021xx)/ ; ZDYF2019073//Key Research and Development Program of Hainan Province (ZDYF2019073)/ ; }, abstract = {Culex quinquefasciatus, one of the most significant mosquito vectors in the world, is widespread in most parts of southern China. A variety of diseases including Bancroft's filariasis, West Nile disease, and St. Louis encephalitis could be transmitted by the vector. Mosquitoes have been shown to host diverse bacterial communities that vary depending on environmental factors such as temperature and rainfall. In this work, 16S rDNA sequencing was used to analyze the seasonal variation of midgut bacterial diversity of Cx. Quinquefasciatus in Haikou City, Hainan Province, China. Proteobacteria was the dominant phylum, accounting for 79.7% (autumn), 73% (winter), 80.4% (spring), and 84.5% (summer). The abundance of Bacteroidetes in autumn and winter was higher than in others. Interestingly, Epsilonbacteraeota, which only exists in autumn and winter, was discovered accidentally in the midgut. We speculated that this might participate in the nutritional supply of adult mosquitoes when temperatures drop. Wolbachia is the most abundant in autumn, accounting for 31.6% of bacteria. The content of Pantoea was highest in the summer group, which might be related to the enhancement of the ability of mosquitoes as temperatures increased. Pseudomonas is carried out as the highest level in winter. On the contrary, in spring and summer, the genus in highest abundance is Enterobacter. Acinetobacter enriches in the spring when it turns from cold to hot. By studying the diversity of midgut bacteria of Cx. quinquefasciatus, we can further understand the co-evolution of mosquitoes and their symbiotic microbes. This is necessary to discuss the seasonal variation of microorganisms and ultimately provide a new perspective for the control of Cx. quinquefasciatus to reduce the spread of the diseases which have notably vital practical significance for the effective prevention of Cx. quinquefasciatus.}, } @article {pmid36005381, year = {2022}, author = {Fallon, AM}, title = {From Mosquito Ovaries to Ecdysone; from Ecdysone to Wolbachia: One Woman's Career in Insect Biology.}, journal = {Insects}, volume = {13}, number = {8}, pages = {}, pmid = {36005381}, issn = {2075-4450}, abstract = {In anautogenous mosquitoes, synchronous development of terminal ovarian follicles after a blood meal provides an important model for studies on insect reproduction. Removal and implantation of ovaries, in vitro culture of dissected tissues and immunological assays for vitellogenin synthesis by the fat body showed that the Aedes aegypti (L.) (Diptera, Culicidae) mosquito ovary produces a factor essential for egg production. The discovery that the ovarian factor was the insect steroid hormone, ecdysone, provided a model for co-option of the larval hormones as reproductive hormones in adult insects. In later work on cultured mosquito cells, ecdysone was shown to arrest the cell cycle, resulting in an accumulation of diploid cells in G1, prior to initiation of DNA synthesis. Some mosquito species, such as Culex pipiens L. (Diptera, Culicidae), harbor the obligate intracellular bacterium, Wolbachia pipientis Hertig (Rickettsiales, Anaplasmataceae), in their reproductive tissues. When maintained in mosquito cell lines, Wolbachia abundance increases in ecdysone-arrested cells. This observation facilitated the recovery of high levels of Wolbachia from cultured cells for microinjection and genetic manipulation. In female Culex pipiens, it will be of interest to explore how hormonal cues that support initiation and progression of the vitellogenic cycle influence Wolbachia replication and transmission to subsequent generations via infected eggs.}, } @article {pmid36005362, year = {2022}, author = {Kyritsis, GA and Koskinioti, P and Bourtzis, K and Papadopoulos, NT}, title = {Effect of Wolbachia Infection and Adult Food on the Sexual Signaling of Males of the Mediterranean Fruit Fly Ceratitis capitata.}, journal = {Insects}, volume = {13}, number = {8}, pages = {}, pmid = {36005362}, issn = {2075-4450}, support = {511 (Coordinated Research Project (CRP) D41025)//International Atomic Energy Agency/ ; HFRI-FM17-4289//Hellenic Foundation for Research and Innovation/ ; }, abstract = {Sexual signaling is a fundamental component of sexual behavior of Ceratitis capitata that highly determines males' mating success. Nutritional status and age are dominant factors known to affect males' signaling performance and define the female decision to accept a male as a sexual partner. Wolbachia pipientis, a widespread endosymbiotic bacterium of insects and other arthropods, exerts several biological effects on its hosts. However, the effects of Wolbachia infection on the sexual behavior of medfly and the interaction between Wolbachia infection and adult food remain unexplored. This study was conducted to determine the effects of Wolbachia on sexual signaling of protein-fed and protein-deprived males. Our findings demonstrate that: (a) Wolbachia infection reduced male sexual signaling rates in both food regimes; (b) the negative effect of Wolbachia infection was more pronounced on protein-fed than protein-deprived males, and it was higher at younger ages, indicating that the bacterium regulates male sexual maturity; (c) Wolbachia infection alters the daily pattern of sexual signaling; and (d) protein deprivation bears significant descent on sexual signaling frequency of the uninfected males, whereas no difference was observed for the Wolbachia-infected males. The impact of our findings on the implementation of Incompatible Insect Technique (IIT) or the combined SIT/IIT towards controlling insect pests is discussed.}, } @article {pmid36003274, year = {2022}, author = {Sharma, M and Kumar, V}, title = {Mosquito-larvicidal Binary (BinA/B) proteins for mosquito control programs -advancements, challenges, and possibilities.}, journal = {Current research in insect science}, volume = {2}, number = {}, pages = {100028}, pmid = {36003274}, issn = {2666-5158}, abstract = {The increasing global burden of mosquito-borne diseases require targeted, environmentally friendly, and sustainable approaches for effective vector control without endangering the non-target beneficial insect population. Biological interventions such as biopesticides, Wolbachia-mediated biological controls, or sterile insect techniques are used worldwide. Here we review Binary or BinAB toxin-the mosquito-larvicidal component of WHO-recognized Lysinibacillus sphaericus bacterium employed in mosquito control programs. Binary (BinAB) toxin is primarily responsible for the larvicidal effect of the bacterium. BinAB is a single-receptor-specific toxin and is effective against larvae of Culex and Anopheles, but not against Aedes aegypti. The receptor in Culex, the Cqm1 protein, has been extensively studied. It is a GPI-anchored amylomaltase and is located apically in the lipid rafts of the larval-midgut epithelium. The interaction of the toxin components with the receptor is crucial for the mosquito larvicidal activity of the BinAB toxin. Here we extend support for the pore formation model of BinAB toxin internalization and the role of toxin-glycan interactions in the endoplasmic reticulum in mediating larval death. BinAB is phylogenetically safe for humans, as Cqm1-like protein is not expected in the human proteome. This review aims to initiate targeted R&D efforts, such as applying fusion technologies (chimera of BinA, chemical modification of BinA), for efficient mosquito control interventions. In addition, the review also examines other areas such as bioremediation and cancer therapeutics, in which L. sphaericus is proving useful and showing potential for further development.}, } @article {pmid36003268, year = {2022}, author = {Davies, OK and Dorey, JB and Stevens, MI and Gardner, MG and Bradford, TM and Schwarz, MP}, title = {Unparalleled mitochondrial heteroplasmy and Wolbachia co-infection in the non-model bee, Amphylaeus morosus.}, journal = {Current research in insect science}, volume = {2}, number = {}, pages = {100036}, pmid = {36003268}, issn = {2666-5158}, abstract = {Mitochondrial heteroplasmy is the occurrence of more than one type of mitochondrial DNA within a single individual. Although generally reported to occur in a small subset of individuals within a species, there are some instances of widespread heteroplasmy across entire populations. Amphylaeus morosus is an Australian native bee species in the diverse and cosmopolitan bee family Colletidae. This species has an extensive geographical range along the eastern Australian coast, from southern Queensland to western Victoria, covering approximately 2,000 km. Seventy individuals were collected from five localities across this geographical range and sequenced using Sanger sequencing for the mitochondrial cytochrome c oxidase subunit I (COI) gene. These data indicate that every individual had the same consistent heteroplasmic sites but no other nucleotide variation, suggesting two conserved and widespread heteroplasmic mitogenomes. Ion Torrent shotgun sequencing revealed that heteroplasmy occurred across multiple mitochondrial protein-coding genes and is unlikely explained by transposition of mitochondrial genes into the nuclear genome (NUMTs). DNA sequence data also demonstrated a consistent co-infection of Wolbachia across the A. morosus distribution with every individual infected with both bacterial strains. Our data are consistent with the presence of two mitogenomes within all individuals examined in this species and suggest a major divergence from standard patterns of mitochondrial inheritance. Because the host's mitogenome and the Wolbachia genome are genetically linked through maternal inheritance, we propose three possible hypotheses that could explain maintenance of the widespread and conserved co-occurring bacterial and mitochondrial genomes in this species.}, } @article {pmid35992686, year = {2022}, author = {Ma, TC and Guo, WJ and Wen, JB}, title = {Effects of feeding on different parts of Ailanthus altissima on the intestinal microbiota of Eucryptorrhynchus scrobiculatus and Eucryptorrhynchus brandti (Coleoptera: Curculionidae).}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {899313}, pmid = {35992686}, issn = {1664-302X}, abstract = {Eucryptorrhynchus brandti and Eucryptorrhynchus scrobiculatus (Coleoptera: Curculionidae) are two monophagous weevil pests that feed on Ailanthus altissima (Mill.) Swingle but differ in their diet niche. In the field, adults of E. brandti prefer to feed on the trunk of A. altissima, whereas adults of E. scrobiculatus prefer to feed on the tender parts. We conducted Illumina sequencing of 16S rRNA to examine changes in bacterial diversity in the adults of these two weevil species after they fed on different parts of A. altissima (trunk, 2-3-year-old branches, annual branches, and petioles). Proteobacteria, Tenericutes, and Firmicutes were the dominant phyla in E. brandti (relative abundance was 50.64, 41.56, and 5.63%, respectively) and E. scrobiculatus (relative abundance was 78.63, 11.91, and 7.41%, respectively). At the genus level, Spiroplasma, endosymbionts2, Unclassified Enterobacteriaceae, and Lactococcus were dominant in E. brandti, and Unclassified Enterobacteriaceae, Wolbachia and Spiroplasma, and endosymbionts2 were dominant in E. scrobiculatus. Linear discriminant analysis effect size analysis revealed microbial biomarkers in the different treatment group of adults of both weevil species. Adults of E. brandti may require the trunk, and adults of E. scrobiculatus may require the petioles and annual branches to maintain the high diversity of their gut microbes. The results of this study indicate that feeding on different parts of A. altissima affects the composition and function of the microbes of E. brandti and the microbial composition of E. scrobiculatus. Variation in the abundance of Wolbachia and Spiroplasma in E. brandti and E. scrobiculatus is associated with dietary niche changes, and this might explain the evolution of reproductive isolation between these two sibling weevil species.}, } @article {pmid35992676, year = {2022}, author = {Sadanandane, C and Gunasekaran, K and Panneer, D and Subbarao, SK and Rahi, M and Vijayakumar, B and Athithan, V and Sakthivel, A and Dinesh, S and Jambulingam, P}, title = {Studies on the fitness characteristics of wMel- and wAlbB-introgressed Aedes aegypti (Pud) lines in comparison with wMel- and wAlbB-transinfected Aedes aegypti (Aus) and wild-type Aedes aegypti (Pud) lines.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {947857}, pmid = {35992676}, issn = {1664-302X}, abstract = {Wolbachia, an intracellular maternally transmitted endosymbiont, has been shown to interfere with the replication of dengue virus in Aedes aegypti mosquitoes. The Wolbachia-transinfected Ae. aegypti has been currently released in many countries to test its effectiveness in preventing the transmission of dengue virus. ICMR-Vector Control Research Centre in collaboration with World Mosquito Program Monash University, Australia, has generated two new Wolbachia-introgressed Ae. aegypti Puducherry (Pud) lines via backcrossing Ae. aegypti females of Australian (Aus) strains, infected with wMel and wAlbB Wolbachia with wild-type Ae. aegypti Puducherry (Pud) males. Wolbachia infections are known to induce a fitness cost and confer benefit on the host mosquito populations that will influence spread of the Wolbachia into native wild mosquito populations during the field release. Hence, the induced fitness cost or benefit/advantage in the two newly generated Ae. aegypti (Pud) lines was assessed in the laboratory in comparison with the wild-type Ae. aegypti (Pud) strain. In addition, maternal transmission (MT) efficiency, induced cytoplasmic incompatibility (CI), and insecticide resistance status of the two (Pud) lines were determined to assess the likely frequency of wMel and wAlbB infections in the native wild population after field invasion. The study shows that wMel and wAlbB infections did not induce any fitness cost on the two newly generated (Pud) lines. Rather, in terms of wing length, fecundity, egg hatch rate, and adult survival, the Wolbachia introgression conferred fitness benefits on the (Pud) lines compared to uninfected Wolbachia free wild Ae. aegypti population. wMel and wAlbB exhibited a high maternal transmission (99-100%) and induced nearly complete (98-100%) cytoplasmic incompatibility. Both the (Pud) lines were resistant to deltamethrin, malathion, DDT, and temephos, and the level of resistance was almost the same between the two lines as in the wild type. Overall, the stable association of wMel and wAlbB established with Ae. aegypti and the reproductive advantages of the (Pud) lines encourage a pilot release in the field for population replacement potential.}, } @article {pmid35976120, year = {2022}, author = {Shastry, V and Bell, KL and Buerkle, CA and Fordyce, JA and Forister, ML and Gompert, Z and Lebeis, SL and Lucas, LK and Marion, ZH and Nice, CC}, title = {A continental-scale survey of Wolbachia infections in blue butterflies reveals evidence of interspecific transfer and invasion dynamics.}, journal = {G3 (Bethesda, Md.)}, volume = {12}, number = {10}, pages = {}, pmid = {35976120}, issn = {2160-1836}, mesh = {Animals ; *Butterflies/genetics/microbiology ; DNA, Mitochondrial/genetics ; Haplotypes/genetics ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Infections by maternally inherited bacterial endosymbionts, especially Wolbachia, are common in insects and other invertebrates but infection dynamics across species ranges are largely under studied. Specifically, we lack a broad understanding of the origin of Wolbachia infections in novel hosts, and the historical and geographical dynamics of infections that are critical for identifying the factors governing their spread. We used Genotype-by-Sequencing data from previous population genomics studies for range-wide surveys of Wolbachia presence and genetic diversity in North American butterflies of the genus Lycaeides. As few as one sequence read identified by assembly to a Wolbachia reference genome provided high accuracy in detecting infections in host butterflies as determined by confirmatory PCR tests, and maximum accuracy was achieved with a threshold of only 5 sequence reads per host individual. Using this threshold, we detected Wolbachia in all but 2 of the 107 sampling localities spanning the continent, with infection frequencies within populations ranging from 0% to 100% of individuals, but with most localities having high infection frequencies (mean = 91% infection rate). Three major lineages of Wolbachia were identified as separate strains that appear to represent 3 separate invasions of Lycaeides butterflies by Wolbachia. Overall, we found extensive evidence for acquisition of Wolbachia through interspecific transfer between host lineages. Strain wLycC was confined to a single butterfly taxon, hybrid lineages derived from it, and closely adjacent populations in other taxa. While the other 2 strains were detected throughout the rest of the continent, strain wLycB almost always co-occurred with wLycA. Our demographic modeling suggests wLycB is a recent invasion. Within strain wLycA, the 2 most frequent haplotypes are confined almost exclusively to separate butterfly taxa with haplotype A1 observed largely in Lycaeides melissa and haplotype A2 observed most often in Lycaeides idas localities, consistent with either cladogenic mode of infection acquisition from a common ancestor or by hybridization and accompanying mutation. More than 1 major Wolbachia strain was observed in 15 localities. These results demonstrate the utility of using resequencing data from hosts to quantify Wolbachia genetic variation and infection frequency and provide evidence of multiple colonizations of novel hosts through hybridization between butterfly lineages and complex dynamics between Wolbachia strains.}, } @article {pmid35967981, year = {2022}, author = {Shropshire, JD and Hamant, E and Conner, WR and Cooper, BS}, title = {cifB-transcript levels largely explain cytoplasmic incompatibility variation across divergent Wolbachia.}, journal = {PNAS nexus}, volume = {1}, number = {3}, pages = {pgac099}, pmid = {35967981}, issn = {2752-6542}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, abstract = {Divergent hosts often associate with intracellular microbes that influence their fitness. Maternally transmitted Wolbachia bacteria are the most common of these endosymbionts, due largely to cytoplasmic incompatibility (CI) that kills uninfected embryos fertilized by Wolbachia-infected males. Closely related infections in females rescue CI, providing a relative fitness advantage that drives Wolbachia to high frequencies. One prophage-associated gene (cifA) governs rescue, and two contribute to CI (cifA and cifB), but CI strength ranges from very strong to very weak for unknown reasons. Here, we investigate CI-strength variation and its mechanistic underpinnings in a phylogenetic context across 20 million years (MY) of Wolbachia evolution in Drosophila hosts diverged up to 50 MY. These Wolbachia encode diverse Cif proteins (100% to 7.4% pairwise similarity), and AlphaFold structural analyses suggest that CifB sequence similarities do not predict structural similarities. We demonstrate that cifB-transcript levels in testes explain CI strength across all but two focal systems. Despite phylogenetic discordance among cifs and the bulk of the Wolbachia genome, closely related Wolbachia tend to cause similar CI strengths and transcribe cifB at similar levels. This indicates that other non-cif regions of the Wolbachia genome modulate cif-transcript levels. CI strength also increases with the length of the host's larval life stage, presumably due to prolonged cif action. Our findings reveal that cifB-transcript levels largely explain CI strength, while highlighting other covariates. Elucidating CI's mechanism contributes to our understanding of Wolbachia spread in natural systems and to improving the efficacy of CI-based biocontrol of arboviruses and agricultural pests globally.}, } @article {pmid35939432, year = {2022}, author = {, }, title = {Retraction: Wolbachia Transcription Elongation Factor "Wol GreA" Interacts with α2ββ'σ Subunits of RNA Polymerase through Its Dimeric C-Terminal Domain.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {8}, pages = {e0010694}, pmid = {35939432}, issn = {1935-2735}, } @article {pmid35925631, year = {2022}, author = {Denton, JA and Joubert, DA and Goundar, AA and Gilles, JRL}, title = {International shipments of Wolbachia-infected mosquito eggs: towards the scaling-up of World Mosquito Program operations.}, journal = {Revue scientifique et technique (International Office of Epizootics)}, volume = {41}, number = {1}, pages = {91-99}, doi = {10.20506/rst.41.1.3306}, pmid = {35925631}, issn = {0253-1933}, mesh = {*Aedes ; Animals ; Australia ; *Dengue/epidemiology/veterinary ; Mosquito Vectors ; Ovum ; Pest Control, Biological ; *Wolbachia ; }, abstract = {The Wolbachia insect control method, employed by the World Mosquito Program (WMP), relies on introgressing Wolbachia through target Aedes aegypti populations to reduce the incidence of dengue. Since 2010, the WMP has been producing Wolbachia-infected mosquitoes at numerous sites across the globe for release in 11 countries. As the technology has matured, greater focus has been placed on mosquito production at larger central facilities for transport to remote release sites, both domestically and internationally. Of particular note is the production of Wolbachia-infected mosquitoes at the WMP's Australian production facility for successful international deployments in Fiji, Vanuatu, Kiribati and Sri Lanka. This requires careful management of both production and supply-chain processes to ensure that the quality of the mosquito eggs, specifically the hatch rate and Wolbachia infection rate, is maintained. To ensure the cost-effectiveness and scalability of the Wolbachia method, these processes will be further refined to facilitate deployment from large centralised production facilities.}, } @article {pmid35923389, year = {2022}, author = {Tibbs-Cortes, LE and Tibbs-Cortes, BW and Schmitz-Esser, S}, title = {Tardigrade Community Microbiomes in North American Orchards Include Putative Endosymbionts and Plant Pathogens.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {866930}, pmid = {35923389}, issn = {1664-302X}, abstract = {The microbiome of tardigrades, a phylum of microscopic animals best known for their ability to survive extreme conditions, is poorly studied worldwide and completely unknown in North America. An improved understanding of tardigrade-associated bacteria is particularly important because tardigrades have been shown to act as vectors of the plant pathogen Xanthomonas campestris in the laboratory. However, the potential role of tardigrades as reservoirs and vectors of phytopathogens has not been investigated further. This study analyzed the microbiota of tardigrades from six apple orchards in central Iowa, United States, and is the first analysis of the microbiota of North American tardigrades. It is also the first ever study of the tardigrade microbiome in an agricultural setting. We utilized 16S rRNA gene amplicon sequencing to characterize the tardigrade community microbiome across four contrasts: location, substrate type (moss or lichen), collection year, and tardigrades vs. their substrate. Alpha diversity of the tardigrade community microbiome differed significantly by location and year of collection but not by substrate type. Our work also corroborated earlier findings, demonstrating that tardigrades harbor a distinct microbiota from their environment. We also identified tardigrade-associated taxa that belong to genera known to contain phytopathogens (Pseudomonas, Ralstonia, and the Pantoea/Erwinia complex). Finally, we observed members of the genera Rickettsia and Wolbachia in the tardigrade microbiome; because these are obligate intracellular genera, we consider these taxa to be putative endosymbionts of tardigrades. These results suggest the presence of putative endosymbionts and phytopathogens in the microbiota of wild tardigrades in North America.}, } @article {pmid35916725, year = {2022}, author = {Dereeper, A and Summo, M and Meyer, DF}, title = {PanExplorer: a web-based tool for exploratory analysis and visualization of bacterial pan-genomes.}, journal = {Bioinformatics (Oxford, England)}, volume = {38}, number = {18}, pages = {4412-4414}, pmid = {35916725}, issn = {1367-4811}, support = {//European Union on the Guadeloupe Region/ ; 2018-FED-1084//European Research and Development Funds/ ; }, mesh = {*Genome, Bacterial ; Genomics ; Software ; *Libraries ; Internet ; }, abstract = {MOTIVATION: As pan-genome approaches are largely employed for bacterial comparative genomics and evolution analyses, but still difficult to be carried out by non-bioinformatician biologists, there is a need for an innovative tool facilitating the exploration of bacterial pan-genomes.

RESULTS: PanExplorer is a web application providing various genomic analyses and reports, giving intuitive views that enable a better understanding of bacterial pan-genomes. As an example, we produced the pan-genome for 121 Anaplasmataceae strains (including 30 Ehrlichia, 15 Anaplasma, 68 Wolbachia).

PanExplorer is written in Perl CGI and relies on several JavaScript libraries for visualization (hotmap.js, MauveViewer, CircosJS). It is freely available at http://panexplorer.southgreen.fr. The source code has been released in a GitHub repository https://github.com/SouthGreenPlatform/PanExplorer. A documentation section is available on PanExplorer website.}, } @article {pmid35916448, year = {2022}, author = {Wang, J and Gou, QY and Luo, GY and Hou, X and Liang, G and Shi, M}, title = {Total RNA sequencing of Phlebotomus chinensis sandflies in China revealed viral, bacterial, and eukaryotic microbes potentially pathogenic to humans.}, journal = {Emerging microbes & infections}, volume = {11}, number = {1}, pages = {2080-2092}, pmid = {35916448}, issn = {2222-1751}, support = {U01 AI151810/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteria/genetics ; Eukaryota/genetics ; Humans ; Mammals ; *Phlebotomus/genetics ; *Phlebovirus/genetics ; *Psychodidae/genetics ; RNA ; Sequence Analysis, RNA ; }, abstract = {Phlebotomus chinensis sandfly is a neglected insect vector in China that is well-known for carrying Leishmania. Recent studies have expanded its pathogen repertoire with two novel arthropod-borne phleboviruses capable of infecting humans and animals. Despite these discoveries, our knowledge of the general pathogen diversity and overall microbiome composition of this vector species is still very limited. Here we carried out a meta-transcriptomics analysis that revealed the actively replicating/transcribing RNA viruses, DNA viruses, bacteria, and eukaryotic microbes, namely, the "total microbiome", of several sandfly populations in China. Strikingly, "microbiome" made up 1.8% of total non-ribosomal RNA and comprised more than 87 species, among which 70 were novel, including divergent members of the genera Flavivirus and of the family Trypanosomatidae. Importantly, among these microbes we were able to reveal four distinguished types of human and/or mammalian pathogens, including two phleboviruses (hedi and wuxiang viruses), one novel Spotted fever group rickettsia, as well as a member of Leishmania donovani complex, among which hedi virus and Leishmania each had > 50% pool prevalence rate and relatively high abundance levels. Our study also showed the ubiquitous presence of an endosymbiont, namely Wolbachia, although no anti-viral or anti-pathogen effects were detected based on our data. In summary, our results uncovered the much un-explored diversity of microbes harboured by sandflies in China and demonstrated that high pathogen diversity and abundance are currently present in multiple populations, implying disease potential for exposed local human population or domestic animals.}, } @article {pmid35914568, year = {2022}, author = {Rosário, AAD and Dias-Lima, AG and Lambert, SM and Souza, BMPDS and Bravo, F}, title = {Identification and molecular characterization of Wolbachia strains and natural infection for Leishmania sp. in neotropical Phlebotominae (Diptera: Psychodidae) species, leishmaniasis vectors.}, journal = {Acta tropica}, volume = {235}, number = {}, pages = {106624}, doi = {10.1016/j.actatropica.2022.106624}, pmid = {35914568}, issn = {1873-6254}, mesh = {Animals ; Brazil ; Insect Vectors/parasitology ; *Leishmania infantum/genetics ; *Leishmaniasis ; *Parasites ; *Psychodidae/parasitology ; *Wolbachia/genetics ; }, abstract = {Recently, Wolbachia infection has been described in leishmaniasis vector sandflies. This endosymbiont bacterium is present in 60% of insects, and has been suggested as a mechanism of biological control of vector insects, because it causes a series of changes in the invertebrate host. In addition, recent studies have shown that this bacterium can prevent the development of parasites in vector insects. In this context, the present study aims to molecularly characterize the circulating strain of this bacterium in sandflies in the State of Bahia, Brazil, as well as the natural infection rate of Leishmania sp., and to evaluate the coinfection between Wolbachia and Leishmania. Seven hundred and forty-five (745) specimens of sandflies were collected in nine municipalities of Bahia, belonging to two species, Lutzomyia longipalpis (Lutz and Neiva, 1912) and Nyssomyia whitmani (Antunes and Coutinho, 1939). The results confirm infection by the protozoan Leishmania infantum and Wolbachia in both species collected. The identified strain of Wolbachia in sandflies was wStv MI, known to lead to a phenotype of cytoplasmic incompatibility in vector insects.}, } @article {pmid35913683, year = {2022}, author = {Su, Y and Zheng, B and Zou, X}, title = {Wolbachia Dynamics in Mosquitoes with Incomplete CI and Imperfect Maternal Transmission by a DDE System.}, journal = {Bulletin of mathematical biology}, volume = {84}, number = {9}, pages = {95}, pmid = {35913683}, issn = {1522-9602}, mesh = {Animals ; *Aedes ; Mathematical Concepts ; Models, Biological ; *Wolbachia ; }, abstract = {In this paper, we propose a delay differential equation model to describe the Wolbachia infection dynamics in mosquitoes in which the key factor of cytoplasmic incompactibility (CI) is incorporated in a more natural way than those in the literature. By analyzing the dynamics of the model, we are able to obtain some information on the impact of four important parameters: the competition capabilities of the wild mosquitoes and infected mosquitoes, the maternal transmission level and the CI level. The analytic results show that there are ranges of parameters that support competition exclusion principle, and there are also ranges of parameters that allow co-persistence for both wild and infected mosquitoes. These ranges account for the scenarios of failure of invasion, invasion and suppressing the wild mosquitoes, and invasion and replacing the wild mosquitoes. We also discuss some possible future problems both in mathematics and in modeling.}, } @article {pmid35895627, year = {2022}, author = {Calle-Tobón, A and Pérez-Pérez, J and Forero-Pineda, N and Chávez, OT and Rojas-Montoya, W and Rúa-Uribe, G and Gómez-Palacio, A}, title = {Local-scale virome depiction in Medellín, Colombia, supports significant differences between Aedes aegypti and Aedes albopictus.}, journal = {PloS one}, volume = {17}, number = {7}, pages = {e0263143}, pmid = {35895627}, issn = {1932-6203}, mesh = {*Aedes/classification/virology ; Animals ; Colombia ; *Insect Viruses/genetics ; Mosquito Vectors/virology ; *RNA Viruses/genetics ; *Virome/genetics ; Wolbachia/genetics ; }, abstract = {Aedes spp. comprise the primary group of mosquitoes that transmit arboviruses such as dengue, Zika, and chikungunya viruses to humans, and thus these insects pose a significant burden on public health worldwide. Advancements in next-generation sequencing and metagenomics have expanded our knowledge on the richness of RNA viruses harbored by arthropods such as Ae. aegypti and Ae. albopictus. Increasing evidence suggests that vector competence can be modified by the microbiome (comprising both bacteriome and virome) of mosquitoes present in endemic zones. Using an RNA-seq-based metataxonomic approach, this study determined the virome structure, Wolbachia presence and mitochondrial diversity of field-caught Ae. aegypti and Ae. albopictus mosquitoes in Medellín, Colombia, a municipality with a high incidence of mosquito-transmitted arboviruses. The two species are sympatric, but their core viromes differed considerably in richness, diversity, and abundance; although the community of viral species identified was large and complex, the viromes were dominated by few virus species. BLAST searches of assembled contigs suggested that at least 17 virus species (16 of which are insect-specific viruses [ISVs]) infect the Ae. aegypti population. Dengue virus 3 was detected in one sample and it was the only pathogenic virus detected. In Ae. albopictus, up to 11 ISVs and one plant virus were detected. Therefore, the virome composition appears to be species-specific. The bacterial endosymbiont Wolbachia was identified in all Ae. albopictus samples and in some Ae. aegypti samples collected after 2017. The presence of Wolbachia sp. in Ae. aegypti was not related to significant changes in the richness, diversity, or abundance of this mosquito's virome, although it was related to an increase in the abundance of Aedes aegypti To virus 2 (Metaviridae). The mitochondrial diversity of these mosquitoes suggested that the Ae. aegypti population underwent a change that started in the second half of 2017, which coincides with the release of Wolbachia-infected mosquitoes in Medellín, indicating that the population of wMel-infected mosquitoes released has introduced new alleles into the wild Ae. aegypti population of Medellín. However, additional studies are required on the dispersal speed and intergenerational stability of wMel in Medellín and nearby areas as well as on the introgression of genetic variants in the native mosquito population.}, } @article {pmid35894613, year = {2022}, author = {Guo, Y and Guo, J and Li, Y}, title = {Wolbachia wPip Blocks Zika Virus Transovarial Transmission in Aedes albopictus.}, journal = {Microbiology spectrum}, volume = {10}, number = {5}, pages = {e0263321}, pmid = {35894613}, issn = {2165-0497}, mesh = {Animals ; Male ; Female ; *Wolbachia/physiology ; *Aedes ; *Zika Virus ; *Zika Virus Infection/prevention & control ; Mosquito Vectors/physiology ; Tetracyclines ; }, abstract = {Wolbachia is being developed as a biological tool to suppress mosquito populations and/or interfere with their transmitted viruses. Adult males with an artificial Wolbachia infection have been released, successfully yielding population suppression in multiple field trials. The main characteristic of the artificial Wolbachia-infected mosquitoes used in the suppression program is the lower vector competence than that in native infected/uninfected mosquitoes in horizontal and vertical transmission. Our previous studies have demonstrated that the Aedes albopictus HC line infected with a trio of Wolbachia strains exhibited almost complete blockade of dengue virus (DENV) and Zika virus (ZIKV) in horizontal and vertical transmission. However, the extent to which Wolbachia inhibits virus transovarial transmission is unknown since no studies have been performed to determine whether Wolbachia protects ovarian cells against viral infection. Here, we employed ovarian cells of the Ae. albopictus GUA (a wild-type mosquito line superinfected with two native Wolbachia strains, wAlbA and wAlbB), HC, and GT lines (tetracycline-cured, Wolbachia-uninfected mosquitoes), which exhibit key traits, and compared them to better understand how Wolbachia inhibits ZIKV transovarial transmission. Our results showed that the infection rate of adult GT progeny was significantly higher than that of GUA progeny during the first and second gonotrophic cycles. In contrast, the infection rates of adult GT and GUA progeny were not significantly different during the third gonotrophic cycle. All examined adult HC progeny from three gonotrophic cycles were negative for ZIKV infection. A strong negative linear correlation existed between Wolbachia density and ZIKV load in the ovaries of mosquitoes. Although there is no obvious coexistence area in the ovaries for Wolbachia and ZIKV, host immune responses may play a role in Wolbachia blocking ZIKV expansion and maintenance in the ovaries of Ae. albopictus. These results will aid in understanding Wolbachia-ZIKV interactions in mosquitoes. IMPORTANCE Area-wide application of Wolbachia to suppress mosquito populations and their transmitted viruses has achieved success in multiple countries. However, the mass release of Wolbachia-infected male mosquitoes involves a potential risk of accidentally releasing fertile females. In this study, we employed ovarian cells of the Ae. albopictus GUA, HC, and GT lines, which exhibit key traits, and compared them to better understand how Wolbachia inhibits ZIKV transovarial transmission. Our results showed an almost complete blockade of ZIKV transmission in HC female mosquitoes. Wolbachia in natively infected GUA mosquitoes negative affected ZIKV, and this interference was shown by slightly lower loads than those in HC mosquitoes. Overall, our work helps show how Wolbachia blocks ZIKV expansion and maintenance in the ovaries of Ae. albopictus and aids in understanding Wolbachia-ZIKV interactions in mosquitoes.}, } @article {pmid35876309, year = {2022}, author = {Schuler, H and Dittmer, J and Borruso, L and Galli, J and Fischnaller, S and Anfora, G and Rota-Stabelli, O and Weil, T and Janik, K}, title = {Investigating the microbial community of Cacopsylla spp. as potential factor in vector competence of phytoplasma.}, journal = {Environmental microbiology}, volume = {24}, number = {10}, pages = {4771-4786}, pmid = {35876309}, issn = {1462-2920}, mesh = {Animals ; *Hemiptera/microbiology ; *Malus/microbiology ; *Microbiota/genetics ; *Phytoplasma/genetics ; Plant Diseases/microbiology ; RNA, Ribosomal, 16S/genetics ; }, abstract = {Phytoplasmas are obligatory intracellular bacteria that colonize the phloem of many plant species and cause hundreds of plant diseases worldwide. In nature, phytoplasmas are primarily transmitted by hemipteran vectors. While all phloem-feeding insects could in principle transmit phytoplasmas, only a limited number of species have been confirmed as vectors. Knowledge about factors that might determine the vector capacity is currently scarce. Here, we characterized the microbiomes of vector and non-vector species of apple proliferation (AP) phytoplasma 'Candidatus Phytoplasma mali' to investigate their potential role in the vector capacity of the host. We performed high-throughput 16S rRNA metabarcoding of the two principal AP-vectors Cacopsylla picta and Cacopsylla melanoneura and eight Cacopsylla species, which are not AP-vectors but co-occur in apple orchards. The microbiomes of all species are dominated by Carsonella, the primary endosymbiont of psyllids and a second uncharacterized Enterobacteriaceae endosymbiont. Each Cacopsylla species harboured a species-specific phylotype of both symbionts. Moreover, we investigated differences between the microbiomes of AP-vector versus non-vector species and identified the predominant endosymbionts but also Wolbachia and several minor taxa as potential indicator species. Our study highlights the importance of considering the microbiome in future investigations of potential factors influencing host vector competence. We investigated the potential role of symbiotic bacteria in the acquisition and transmission of phytoplasma. By comparing the two main psyillid vector species of Apple proliferation (AP) phytoplasma and eight co-occurring species, which are not able to vector AP-phytoplasma, we found differences in the microbial communities of AP-vector and non-vector species, which appear to be driven by the predominant symbionts in both vector species and Wolbachia and several minor taxa in the non-vector species. In contrast, infection with AP-phytoplasma did not affect microbiome composition in both vector species. Our study provides new insights into the endosymbiont diversity of Cacopsylla spp. and highlights the importance of considering the microbiome when investigating potential factors influencing host vector competence.}, } @article {pmid35876244, year = {2022}, author = {Sawadogo, SP and Kabore, DA and Tibiri, EB and Hughes, A and Gnankine, O and Quek, S and Diabaté, A and Ranson, H and Hughes, GL and Dabiré, RK}, title = {Lack of robust evidence for a Wolbachia infection in Anopheles gambiae from Burkina Faso.}, journal = {Medical and veterinary entomology}, volume = {36}, number = {3}, pages = {301-308}, pmid = {35876244}, issn = {1365-2915}, support = {MR/P027873/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; *Anopheles/genetics ; Burkina Faso ; *Malaria/veterinary ; Mosquito Vectors ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {The endosymbiont Wolbachia can have major effects on the reproductive fitness, and vectorial capacity of host insects and may provide new avenues to control mosquito-borne pathogens. Anopheles gambiae s.l is the major vector of malaria in Africa but the use of Wolbachia in this species has been limited by challenges in establishing stable transinfected lines and uncertainty around native infections. High frequencies of infection of Wolbachia have been previously reported in An. gambiae collected from the Valle du Kou region of Burkina Faso in 2011 and 2014. Here, we re-evaluated the occurrence of Wolbachia in natural samples, collected from Valle du Kou over a 12-year time span, and in addition, expanded sampling to other sites in Burkina Faso. Our results showed that, in contrast to earlier reports, Wolbachia is present at an extremely low prevalence in natural population of An. gambiae. From 5341 samples analysed, only 29 were positive for Wolbachia by nested PCR representing 0.54% of prevalence. No positive samples were found with regular PCR. Phylogenetic analysis of 16S rRNA gene amplicons clustered across supergroup B, with some having similarity to sequences previously found in Anopheles from Burkina Faso. However, we cannot discount the possibility that the amplicon positive samples we detected were due to environmental contamination or were false positives. Regardless, the lack of a prominent native infection in An. gambiae s.l. is encouraging for applications utilizing Wolbachia transinfected mosquitoes for malaria control.}, } @article {pmid35875588, year = {2022}, author = {Chun, SJ and Cui, Y and Yoo, SH and Lee, JR}, title = {Organic Connection of Holobiont Components and the Essential Roles of Core Microbes in the Holobiont Formation of Feral Brassica napus.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {920759}, pmid = {35875588}, issn = {1664-302X}, abstract = {Brassica napus (Rapeseed) is an econfomically important oil-producing crop. The microbial interactions in the plant holobiont are fundamental to the understanding of plant growth and health. To investigate the microbial dynamics in the holobiont of feral B. napus, a total of 215 holobiont samples, comprised of bulk soil, primary root, lateral root, dead leaf, caulosphere, basal leaf, apical leaf, carposphere, and anthosphere, were collected from five different grassland sites in South Korea. The soil properties differed in different sampling sites, but prokaryotic communities were segregated according to plant holobiont components. The structures of the site-specific SparCC networks were similar across the regions. Recurrent patterns were found in the plant holobionts in the recurrent network. Ralstonia sp., Massilia sp., and Rhizobium clusters were observed consistently and were identified as core taxa in the phyllosphere, dead leaf microbiome, and rhizosphere, respectively. Arthropod-related microbes, such as Wolbachia sp., Gilliamella sp., and Corynebacteriales amplicon sequence variants, were found in the anthosphere. PICRUSt2 analysis revealed that microbes also possessed specific functions related to holobiont components, such as functions related to degradation pathways in the dead leaf microbiome. Structural equation modeling analysis showed the organic connections among holobiont components and the essential roles of the core microbes in the holobiont formations in natural ecosystem. Microbes coexisting in a specific plant showed relatively stable community structures, even though the regions and soil characteristics were different. Microbes in each plant component were organically connected to form their own plant holobiont. In addition, plant-related microbes, especially core microbes in each holobiont, showed recurrent interaction patterns that are essential to an understanding of the survival and coexistence of plant microbes in natural ecosystems.}, } @article {pmid35873163, year = {2022}, author = {Ramos, LFC and Martins, M and Murillo, JR and Domont, GB and de Oliveira, DMP and Nogueira, FCS and Maciel-de-Freitas, R and Junqueira, M}, title = {Interspecies Isobaric Labeling-Based Quantitative Proteomics Reveals Protein Changes in the Ovary of Aedes aegypti Coinfected With ZIKV and Wolbachia.}, journal = {Frontiers in cellular and infection microbiology}, volume = {12}, number = {}, pages = {900608}, pmid = {35873163}, issn = {2235-2988}, mesh = {*Aedes/microbiology ; Animals ; *Coinfection ; Female ; Humans ; Infant, Newborn ; Mosquito Vectors ; Ovary ; Proteomics ; *Wolbachia ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {Zika is a vector-borne disease caused by an arbovirus (ZIKV) and overwhelmingly transmitted by Ae. aegypti. This disease is linked to adverse fetal outcomes, mostly microcephaly in newborns, and other clinical aspects such as acute febrile illness and neurologic complications, for example, Guillain-Barré syndrome. One of the most promising strategies to mitigate arbovirus transmission involves releasing Ae. aegypti mosquitoes carrying the maternally inherited endosymbiont bacteria Wolbachia pipientis. The presence of Wolbachia is associated with a reduced susceptibility to arboviruses and a fitness cost in mosquito life-history traits such as fecundity and fertility. However, the mechanisms by which Wolbachia influences metabolic pathways leading to differences in egg production remains poorly known. To investigate the impact of coinfections on the reproductive tract of the mosquito, we applied an isobaric labeling-based quantitative proteomic strategy to investigate the influence of Wolbachia wMel and ZIKV infection in Ae. aegypti ovaries. To the best of our knowledge, this is the most complete proteome of Ae. aegypti ovaries reported so far, with a total of 3913 proteins identified, were also able to quantify 1044 Wolbachia proteins in complex sample tissue of Ae. aegypti ovary. Furthermore, from a total of 480 mosquito proteins modulated in our study, we discuss proteins and pathways altered in Ae. aegypti during ZIKV infections, Wolbachia infections, coinfection Wolbachia/ZIKV, and compared with no infection, focusing on immune and reproductive aspects of Ae. aegypti. The modified aspects mainly were related to the immune priming enhancement by Wolbachia presence and the modulation of the Juvenile Hormone pathway caused by both microorganism's infection.}, } @article {pmid35869302, year = {2022}, author = {Mejia, AJ and Jimenez, L and Dutra, HLC and Perera, R and McGraw, EA}, title = {Attempts to use breeding approaches in Aedes aegypti to create lines with distinct and stable relative Wolbachia densities.}, journal = {Heredity}, volume = {129}, number = {4}, pages = {215-224}, pmid = {35869302}, issn = {1365-2540}, support = {R01 AI151166/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes/genetics ; Animals ; Mosquito Vectors/genetics ; Specific Gravity ; Virus Replication ; *Wolbachia/genetics ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {Wolbachia is an insect endosymbiont being used for biological control in the mosquito Aedes aegypti because it causes cytoplasmic incompatibility (CI) and limits viral replication of dengue, chikungunya, and Zika viruses. While the genetic mechanism of pathogen blocking (PB) is not fully understood, the strength of both CI and PB are positively correlated with Wolbachia densities in the host. Wolbachia densities are determined by a combination of Wolbachia strain and insect genotype, as well as interactions with the environment. We employed both artificial selection and inbreeding with the goal of creating lines of Ae. aegypti with heritable and distinct Wolbachia densities so that we might better dissect the mechanism underlying PB. We were unable to shift the mean relative Wolbachia density in Ae. aegypti lines by either strategy, with relative densities instead tending to cycle over a narrow range. In lieu of this, we used Wolbachia densities in mosquito legs as predictors of relative densities in the remaining individual's carcass. Because we worked with outbred mosquitoes, our findings indicate either a lack of genetic variation in the mosquito for controlling relative density, natural selection against extreme densities, or a predominance of environmental factors affecting densities. Our study reveals that there are moderating forces acting on relative Wolbachia densities that may help to stabilize density phenotypes post field release. We also show a means to accurately bin vector carcasses into high and low categories for non-DNA omics-based studies of Wolbachia-mediated traits.}, } @article {pmid35867318, year = {2022}, author = {Sugimoto, TN and Watanabe, K and Akiduki, G and Imanishi, S and Mitsuhashi, W}, title = {A new continuous cell line from the pest insect, Anomala cuprea (Coleoptera; Scarabaeidae): emergence of contractile cells.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {58}, number = {7}, pages = {610-618}, pmid = {35867318}, issn = {1543-706X}, mesh = {Animals ; Cell Culture Techniques ; Cell Line ; *Coleoptera ; }, abstract = {Insect contractile cells frequently appear at an early phase of cell culture, but in most cases, they disappear before a continuous cell line is established, so the cell line ceases to contract. Continuous contractile insect cell lines are currently available from only one species each of Hymenoptera and Diptera. Here, we obtained a new cell line that contracted long after being established as a continuous cell line. The cell line contracted for a short period at an early phase of insect cell culture before a continuous cell line was established, but then did not contract again for several years. After this cell line entered the continuous growth phase, it produced spontaneously contractile tissues for about 4 mo but stopped contracting again. This is the first instance of a cell line that contracted after its establishment as a non-contractile continuous cell line. It is unclear whether the contractile cells survive or die after contraction ceases at an early phase of cell culture, and our results indicate that potential contractile cells survive for years after they stop to contract. The cells of this line sometimes produced complex contractile structures, such as sheet-like tissues. Only a few continuous cell lines have been derived from scarabaeid beetles. The new continuous cell line was derived from the culture of the fat bodies of the scarab beetle Anomala cuprea, which is a pest in the agriculture and forestry of Japan. The population doubling time of the new cell line was 2.5 d and thus it grows very rapidly among coleopteran continuous cell lines. Our new cell line will facilitate research on the physiology and pathology of Coleoptera, including scarab beetles, and may also contribute to research on invertebrate muscles.}, } @article {pmid35862784, year = {2022}, author = {Voronin, D and Makepeace, BL}, title = {Symbionts on the Brain: How Wolbachia Is Strictly Corralled in Some Neotropical Drosophila spp.}, journal = {mBio}, volume = {13}, number = {4}, pages = {e0118222}, pmid = {35862784}, issn = {2150-7511}, mesh = {Animals ; Brain ; Drosophila/microbiology ; Female ; *Nematoda ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia is a heritable alphaproteobacterial symbiont of arthropods and nematodes, famous for its repertoire of host manipulations, including cytoplasmic incompatibility. To be vertically transmitted, Wolbachia must efficiently colonize the female germ line, although somatic tissues outside the gonads are also infected. In Drosophila spp., Wolbachia is usually distributed systemically in multiple regions of the adult fly, but in some neotropical hosts, Wolbachia's only somatic niches are cerebral bacteriocyte-like structures and the ovarian follicle cells. In their recent article, Strunov and colleagues (A. Strunov, K. Schmidt, M. Kapun, and W. J. Miller. mBio 13:e03863-21, 2022, https://doi.org/10.1128/mbio.03863-21) compared the development of Drosophila spp. with systemic or restricted infections and demonstrated that the restricted pattern is determined in early embryogenesis by an apparently novel autophagic process, involving intimate interactions of Wolbachia with the endoplasmic reticulum. This work has implications not only for the evolution of neotropical Drosophila spp. but also for our understanding of how Wolbachia infections are controlled in other native or artificial hosts.}, } @article {pmid35859330, year = {2022}, author = {Wang, W and Cui, W and Yang, H}, title = {Toward an accurate mechanistic understanding of Wolbachia-induced cytoplasmic incompatibility.}, journal = {Environmental microbiology}, volume = {24}, number = {10}, pages = {4519-4532}, doi = {10.1111/1462-2920.16125}, pmid = {35859330}, issn = {1462-2920}, mesh = {Animals ; Cytoplasm/genetics ; Cytosol ; Female ; Male ; Phylogeny ; Reproduction ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are the most successful intracellular bacteria in arthropods. They can manipulate host reproduction to favour infected females, which transmit Wolbachia to their progeny and increase the presence of Wolbachia in the population. The reproductive alterations caused by Wolbachia include feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI), among which CI is the most common. CI leads to embryonic lethality when Wolbachia-infected males mate with uninfected females or those infected with an incompatible strain. This lethality can be rescued if females are infected with a compatible strain. Although CI was described in the 1960s and its connection to Wolbachia was made in the 1970s, the genes responsible for CI, called CI factors, were not identified until recently. Since then, significant progress has been made in understanding the molecular mechanism of CI using a combination of genetic, phylogenetic, biochemical and structural approaches. The detailed molecular mechanisms behind this fascinating endosymbiotic bacteria-induced phenotype have begun to emerge. Here, we summarize recent progress in understanding the molecular mechanism of CI, especially focusing on the recently solved CI factor structures and discussing what these new structures brought in terms of CI mechanism.}, } @article {pmid35857094, year = {2022}, author = {Rau, J and Werner, D and Beer, M and Höper, D and Kampen, H}, title = {The microbial RNA metagenome of Aedes albopictus (Diptera: Culicidae) from Germany.}, journal = {Parasitology research}, volume = {121}, number = {9}, pages = {2587-2599}, pmid = {35857094}, issn = {1432-1955}, mesh = {*Aedes ; Animals ; Female ; Humans ; Introduced Species ; Male ; Metagenome ; Mosquito Vectors ; RNA ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {Aedes albopictus is a highly invasive mosquito species that has become widespread across the globe. In addition, it is an efficient vector of numerous pathogens of medical and veterinary importance, including dengue, chikungunya and Zika viruses. Among others, the vector potential of mosquitoes is influenced by their microbiome. However, this influence is very dynamic and can vary between individuals and life stages. To obtain a rough overview on the microbiome of Ae. albopictus populations in Germany, pooled female and pooled male individuals from seven German locations were investigated by total RNA sequencing. The mosquito specimens had been collected as larvae in the field and processed immediately after adult emergence, i.e. without females having fed on blood. RNA fragments with high degrees of identity to a large number of viruses and microorganisms were identified, including, for example, Wolbachia pipientis and Acinetobacter baumannii, with differences between male and female mosquitoes. Knowledge about the natural occurrence of microorganisms in mosquitoes may be translated into new approaches to vector control, for example W. pipientis can be exploited to manipulate mosquito reproduction and vector competence. The study results show how diverse the microbiome of Ae. albopictus can be, and the more so needs to be adequately analysed and interpreted.}, } @article {pmid35853080, year = {2022}, author = {Karimian, F and Koosha, M and Choubdar, N and Oshaghi, MA}, title = {Comparative analysis of the gut microbiota of sand fly vectors of zoonotic visceral leishmaniasis (ZVL) in Iran; host-environment interplay shapes diversity.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {7}, pages = {e0010609}, pmid = {35853080}, issn = {1935-2735}, mesh = {Animals ; Bacteria/genetics ; *Gastrointestinal Microbiome ; Humans ; Iran/epidemiology ; *Leishmaniasis, Visceral/epidemiology ; *Phlebotomus/parasitology ; *Psychodidae/parasitology ; RNA, Ribosomal, 16S/genetics ; }, abstract = {The development of Leishmania parasites within sand fly vectors occurs entirely in the insect gut lumen, in the presence of symbiotic and commensal bacteria. The impacts of host species and environment on the gut microbiome are currently poorly understood. We employed MiSeq sequencing of the V3-16S rRNA gene amplicons to characterize and compare the gut microbiota of field-collected populations of Phlebotomus kandelakii, P. perfiliewi, P. alexandri, and P. major, the primary or secondary vectors of zoonotic visceral leishmaniasis (ZVL) in three distinct regions of Iran where ZVL is endemic. In total, 160,550 quality-filtered reads of the V3 region yielded a total of 72 operational taxonomic units (OTUs), belonging to 23 phyla, 47 classes, 91 orders, 131 families, and 335 genera. More than 50% of the bacteria identified were Proteobacteria, followed by Firmicutes (22%), Deinococcus-Thermus (9%), Actinobacteria (6%), and Bacteroidetes (5%). The core microbiome was dominated by eight genera: Acinetobacter, Streptococcus, Enterococcus, Staphylococcus, Bacillus, Propionibacterium, Kocuria, and Corynebacterium. Wolbachia were found in P. alexandri and P. perfiliewi, while Asaia sp. was reported in P. perfiliewi. Substantial variations in the gut bacterial composition were found between geographically distinct populations of the same sand fly species, as well as between different species at the same location, suggesting that sand fly gut microbiota is shaped by both the host species and geographical location. Phlebotomus kandelakii and P. perfiliewi in the northwest, and P. alexandri in the south, the major ZVL vectors, harbor the highest bacterial diversity, suggesting a possible relationship between microbiome diversity and the capacity for parasite transmission. In addition, large numbers of gram-positive human or animal pathogens were found, suggesting that sand fly vectors of ZVL could pose a potential additional threat to livestock and humans in the region studied. The presence of Bacillus subtilis, Enterobacter cloacae, and Asaia sp suggests that these bacteria could be promising candidates for a paratransgenesis approach to the fight against Leishmaniasis.}, } @article {pmid35849008, year = {2022}, author = {Chaves, EB and Nascimento-Pereira, AC and Pinto, JLM and Rodrigues, BL and de Andrade, MS and Rêbelo, JMM}, title = {Detection of Wolbachia in Mosquitoes (Diptera: Culicidae) in the State of Maranhão, Brazil.}, journal = {Journal of medical entomology}, volume = {59}, number = {5}, pages = {1831-1836}, doi = {10.1093/jme/tjac092}, pmid = {35849008}, issn = {1938-2928}, mesh = {*Aedes/microbiology ; Animals ; *Anopheles/microbiology ; Brazil ; *Culex/microbiology ; *Culicidae/microbiology ; *Wolbachia ; }, abstract = {Recently, the endobacteria Wolbachia has emerged as a biological tool for the control of arboviruses. Thus, we investigated the rate of natural infection by Wolbachia in Culicidae species from Maranhão, Brazil. For this, we amplified the Wolbachia surface protein gene (wsp) from mosquitoes collected in six localities of Maranhão, and positive samples were subjected to new analysis using group-specific primers. In total, 448 specimens comprising 6 genera and 18 species of mosquitoes were analyzed. Wolbachia DNA was PCR-detected in 7 species, three of which are new records: Aedes scapularis (Rondani, 1848), Coquillettidia juxtamansonia (Chagas, 1907) and Cq. venezuelensis (Theobald, 1912), in addition to Ae. albopictus (Skuse, 1894) and Culex quinquefasciatus Say, 1823, which are commonly described as permissive to maintain this bacterium in natural environments, and two species of the subgenera Anopheles (Nyssorhynchus) Blanchard, 1902 and Culex (Melanoconion) Theobald, 1903 which could not be identified at species level. The infection rate of all species ranged from 0 to 80%, and the average value was 16.5%. This study increases the knowledge about the prevalence of Wolbachia in the culicid fauna and may help in selecting strains for biological control purposes.}, } @article {pmid35840683, year = {2023}, author = {Hubert, J and Nesvorna, M and Bostlova, M and Sopko, B and Green, SJ and Phillips, TW}, title = {The Effect of Residual Pesticide Application on Microbiomes of the Storage Mite Tyrophagus putrescentiae.}, journal = {Microbial ecology}, volume = {85}, number = {4}, pages = {1527-1540}, pmid = {35840683}, issn = {1432-184X}, support = {21-337-J//Kansas State Research and Extension/ ; LTAUSA19012//Ministerstvo Školství, Mládeže a Tělovýchovy/ ; }, mesh = {Animals ; *Acaridae/microbiology ; *Pesticides/pharmacology ; *Pesticide Residues/pharmacology ; *Microbiota ; *Mites/microbiology ; *Bacillus/genetics ; *Bartonella ; *Pyrethrins/pharmacology ; }, abstract = {Arthropods can host well-developed microbial communities, and such microbes can degrade pesticides and confer tolerance to most types of pests. Two cultures of the stored-product mite Tyrophagus putrescentiae, one with a symbiotic microbiome containing Wolbachia and the other without Wolbachia, were compared on pesticide residue (organophosphate: pirimiphos-methyl and pyrethroid: deltamethrin, deltamethrin + piperonyl butoxide)-containing diets. The microbiomes from mite bodies, mite feces and debris from the spent mite diet were analyzed using barcode sequencing. Pesticide tolerance was different among mite cultures and organophosphate and pyrethroid pesticides. The pesticide residues influenced the microbiome composition in both cultures but without any remarkable trend for mite cultures with and without Wolbachia. The most influenced bacterial taxa were Bartonella-like and Bacillus for both cultures and Wolbachia for the culture containing this symbiont. However, there was no direct evidence of any effect of Wolbachia on pesticide tolerance. The high pesticide concentration residues in diets reduced Wolbachia, Bartonella-like and Bacillus in mites of the symbiotic culture. This effect was low for Bartonella-like and Bacillus in the asymbiotic microbiome culture. The results showed that the microbiomes of mites are affected by pesticide residues in the diets, but the effect is not systemic. No actual detoxification effect by the microbiome was observed for the tested pesticides.}, } @article {pmid35830425, year = {2022}, author = {Wang, D and Zhang, Y and Xu, M and Sun, X and Cui, X and Wang, X and Liu, D}, title = {Dietary Bacillus licheniformis improves the effect of Astragalus membranaceus extract on blood glucose by regulating antioxidation activity and intestinal microbiota in InR[E19]/TM2 Drosophila melanogaster.}, journal = {PloS one}, volume = {17}, number = {7}, pages = {e0271177}, pmid = {35830425}, issn = {1932-6203}, mesh = {Animals ; Antioxidants/pharmacology ; Astragalus propinquus ; *Bacillus licheniformis/physiology ; Blood Glucose ; Diet ; Dietary Supplements/analysis ; Drosophila melanogaster/microbiology ; *Gastrointestinal Microbiome ; Male ; }, abstract = {BACKGROUND: The diabetes mellitus prevalence is rapidly increasing in most parts of the world and has become a vital health problem. Probiotic and herbal foods are valuable in the treatment of diabetes.

METHODS AND PERFORMANCE: In this study, Bacillus licheniformis (BL) and Astragalus membranaceus extract (AE) were given with food to InR[E19]/TM2 Drosophila melanogaster, and the blood glucose, antioxidation activity and intestinal microbiota were investigated. The obtained results showed that BA (BL and AE combination) supplementation markedly decreased the blood glucose concentration compared with the standard diet control group, accompanied by significantly increased enzymatic activities of catalase (CAT), decreased MDA levels and prolonged lifespan of InR[E19]/TM2 D. melanogaster. The treatments with BL, AE and BA also ameliorated intestinal microbiota equilibrium by increasing the population of Lactobacillus and significantly decreasing the abundance of Wolbachia. In addition, clearly different evolutionary clusters were found among the control, BL, AE and BA-supplemented diets, and the beneficial microbiota, Lactobacillaceae and Acetobacter, were found to be significantly increased in male flies that were fed BA. These results indicated that dietary supplementation with AE combined with BL not only decreased blood glucose but also extended the lifespan, with CAT increasing, MDA decreasing, and intestinal microbiota improving in InR[E19]/TM2 D. melanogaster.

CONCLUSION: The obtained results showed that dietary supplementation with BL and AE, under the synergistic effect of BL and AE, not only prolonged the lifespan of InR[E19]/TM2 D. melanogaster, increased body weight, and improved the body's antiaging enzyme activity but also effectively improved the types and quantities of beneficial bacteria in the intestinal flora of InR[E19]/TM2 D. melanogaster to improve the characteristics of diabetes symptoms. This study provides scientific evidence for a safe and effective dietary therapeutic method for diabetes mellitus.}, } @article {pmid35814684, year = {2022}, author = {Barman, M and Samanta, S and Upadhyaya, G and Thakur, H and Chakraborty, S and Samanta, A and Tarafdar, J}, title = {Unraveling the Basis of Neonicotinoid Resistance in Whitefly Species Complex: Role of Endosymbiotic Bacteria and Insecticide Resistance Genes.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {901793}, pmid = {35814684}, issn = {1664-302X}, abstract = {Bemisia tabaci (whitefly) is one of the most detrimental agricultural insect pests and vectors of many plant viruses distributed worldwide. Knowledge of the distribution patterns and insecticide resistance of this cryptic species is crucial for its management. In this study, genetic variation of mitochondrial cytochrome oxidase subunit 1 (MtCoI) gene of B. tabaci was analyzed followed by a study of the infection profile of various endosymbionts in 26 whitefly populations collected from West Bengal, India. Phylogenetic analysis revealed Asia I as the major cryptic species (65.38%), followed by Asia II 5, China 3, and Asia II 7, which were diversified into 20 different haplotypes. In addition to the primary endosymbiont (C. poriera), each of the four whitefly species showed a variable population of three secondary endosymbionts, majorly Arsenophonus with the highest infection rate (73.07%), followed by Wolbachia and Rickettsia. Further phylogenetic analyses revealed the presence of two subgroups of Arsenophonus, viz., A1 and A2, and one each in Wolbachia (W1) and Rickettsia (R3). Resistance to thiamethoxam, imidacloprid, and acetamiprid insecticides was analyzed for a clear picture of pesticide resistance status. The highest susceptibility was noted toward thiamethoxam (LC50 = 5.36 mg/L), followed by imidacloprid and acetamiprid. The whitefly population from Purulia and Hooghly districts bearing Asia II 7 and Asia II 5 cryptic species, respectively, shows maximum resistance. The differences in mean relative titer of four symbiotic bacteria among field populations varied considerably; however, a significant positive linear correlation was observed between the resistance level and relative titer of Arsenophonus and Wolbachia in the case of imidacloprid and thiamethoxam, while only Wolbachia was found in case of acetamiprid. Expression analysis demonstrated differential upregulation of insecticide resistance genes with Purulia and Hooghly populations showing maximally upregulated P450 genes. Moreover, thiamethoxam and imidacloprid resistance ratio (RR) showed a significant correlation with CYP6CM1, CYP6DZ7, and CYP4C64 genes, while acetamiprid RR correlated with CYP6CX1, CYP6DW2, CYP6DZ7, and CYP4C64 genes. Taken together, these findings suggested that P450 mono-oxygenase and symbiotic bacteria together affected whitefly resistance to neonicotinoids. Hence, a symbiont-oriented management programme could be a better alternative to control or delay resistance development in whitefly and can be used for pesticide clean-up in an agricultural field.}, } @article {pmid35802173, year = {2023}, author = {Rocha, FP and Ronque, MUV and Lyra, ML and Bacci, M and Oliveira, PS}, title = {Habitat and Host Species Drive the Structure of Bacterial Communities of Two Neotropical Trap-Jaw Odontomachus Ants : Habitat and Host Species Drive the Structure of Bacterial Communities of Two Neotropical Trap-Jaw Odontomachus Ants.}, journal = {Microbial ecology}, volume = {86}, number = {1}, pages = {699-712}, pmid = {35802173}, issn = {1432-184X}, support = {Finance Code 001//Coordenação de Aperfeiçoamento de Pessoal de Nível Superior/ ; 2017/26162-8//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2017/04824-9//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2014/23141-1//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2017/16645-1//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 163546/2020-7//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 441036/2016-3//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 307541/2018-5//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 306115/2013-1//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 302219/2017-0//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 303730/2021-8//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; }, mesh = {Animals ; *Ants ; RNA, Ribosomal, 16S/genetics ; Ecosystem ; Brazil ; Bacteria/genetics ; }, abstract = {Ants have long been known for their associations with other taxa, including macroscopic fungi and symbiotic bacteria. Recently, many ant species have had the composition and function of their bacterial communities investigated. Due to its behavioral and ecological diversity, the subfamily Ponerinae deserves more attention regarding its associated microbiota. Here, we used the V4 region of the 16S rRNA gene to characterize the bacterial communities of Odontomachus chelifer (ground-nesting) and Odontomachus hastatus (arboreal), two ponerine trap-jaw species commonly found in the Brazilian savanna ("Cerrado") and Atlantic rainforest. We investigated habitat effects (O. chelifer in the Cerrado and the Atlantic rainforest) and species-specific effects (both species in the Atlantic rainforest) on the bacterial communities' structure (composition and abundance) in two different body parts: cuticle and gaster. Bacterial communities differed in all populations studied. Cuticular communities were more diverse, while gaster communities presented variants common to other ants, including Wolbachia and Candidatus Tokpelaia hoelldoblerii. Odontomachus chelifer populations presented different communities in both body parts, highlighting the influence of habitat type. In the Atlantic rainforest, the outcome depended on the body part targeted. Cuticular communities were similar between species, reinforcing the habitat effect on bacterial communities, which are mainly composed of environmentally acquired taxa. Gaster communities, however, differed between the two Odontomachus species, suggesting species-specific effects and selective filters. Unclassified Firmicutes and uncultured Rhizobiales variants are the main components accounting for the observed differences. Our study indicates that both host species and habitat act synergistically, but to different degrees, to shape the bacterial communities in these Odontomachus species.}, } @article {pmid35799282, year = {2022}, author = {Gharabigloozare, Y and Bleidorn, C}, title = {Effect of high temperature on Wolbachia density and impact on cytoplasmic incompatibility in confused flour beetle, Tribolium confusum (Coleoptera: Tenebrionidae).}, journal = {BMC research notes}, volume = {15}, number = {1}, pages = {240}, pmid = {35799282}, issn = {1756-0500}, mesh = {Animals ; *Coleoptera ; Cytoplasm ; Temperature ; *Tribolium ; *Wolbachia ; }, abstract = {OBJECTIVES: Environmental constraints, especially temperature, have been identified as a key in understanding host-symbiont relationships, as they can directly impact the fitness of the symbiont population and the host development. Here we investigated the effect of temperature during the host development on the density of intracellular bacteria of the Wolbachia, wTcon strain within the confused flour beetle, Tribolium confusum. The wTcon can induce a complete cytoplasmic incompatibility (CI) in T. confusum beetles; therefore, we observed the effect of heat stress on the symbiont-mediated CI.

RESULTS: The density of CI inducing Wolbachia in the Tribolium confusum is temperature-specific. Our observation of the beetles reared in five different temperatures (30-34 °C) measured the highest Wolbachia density at 30-31 °C and lowest at 34 °C within a single insect generation. In this species, changes in the density of Wolbachia related to higher temperature did not influence CI. However, the fertility of beetles reared in higher temperatures showed a substantial decrease in the number of laid and hatched eggs. Thus, we can confirm the effect of high temperature on lowering the wTcon density and no impact on induction of cytoplasmic incompatibility (CI) in T. confusum beetles.}, } @article {pmid35784453, year = {2022}, author = {Wybouw, N and Mortier, F and Bonte, D}, title = {Interacting host modifier systems control Wolbachia-induced cytoplasmic incompatibility in a haplodiploid mite.}, journal = {Evolution letters}, volume = {6}, number = {3}, pages = {255-265}, pmid = {35784453}, issn = {2056-3744}, abstract = {Reproductive parasites such as Wolbachia spread within host populations by inducing cytoplasmic incompatibility (CI). CI occurs when parasite-modified sperm fertilizes uninfected eggs and is typified by great variation in strength across biological systems. In haplodiploid hosts, CI has different phenotypic outcomes depending on whether the fertilized eggs die or develop into males. Genetic conflict theories predict the evolution of host modulation of CI, which in turn influences the stability of reproductive parasitism. However, despite the ubiquity of CI-inducing parasites in nature, there is scarce evidence for intraspecific host modulation of CI strength and phenotype. Here, we tested for intraspecific host modulation of Wolbachia-induced CI in haplodiploid Tetranychus urticae mites. Using a single CI-inducing Wolbachia variant and mitochondrion, a nuclear panel was created that consisted of infected and cured near-isogenic lines. We performed a highly replicated age-synchronized full diallel cross composed of incompatible and compatible control crosses. We uncovered host modifier systems that cause striking variation in CI strength when carried by infected T. urticae males. We observed a continuum of CI phenotypes in our crosses and identified strong intraspecific female modulation of the CI phenotype. Crosses established a recessive genetic basis for the maternal effect and were consistent with polygenic Mendelian inheritance. Both male and female modulation interacted with the genotype of the mating partner. Our findings identify spermatogenesis as an important target of selection for host modulation of CI strength and underscore the importance of maternal genetic effects for the CI phenotype. Our findings reveal that intraspecific host modulation of CI is underpinned by complex genetic architectures and confirm that the evolution of reproductive parasitism is contingent on host genetics.}, } @article {pmid35767080, year = {2022}, author = {Hong, YH and Mai, ZH and Li, CJ and Zheng, QY and Guo, LX}, title = {Microbial Diversity Analyses of Fertilized Thitarodes Eggs and Soil Provide New Clues About the Occurrence of Chinese Cordyceps.}, journal = {Current microbiology}, volume = {79}, number = {8}, pages = {229}, pmid = {35767080}, issn = {1432-0991}, support = {81303155//National Natural Science Foundation of China/ ; 2018A030313094//Natural Science Foundation of Guangdong Province/ ; 2020A151501457//Natural Science Foundation of Guangdong Province/ ; 2018GkQNCX050//Project for Young Innovative Talents in Ordinary Higher University of Guangdong Province/ ; 4SG19057G//Group-type Special Supporting Project for Educational Talents in Universities/ ; }, mesh = {Animals ; Bacteria/genetics ; *Cordyceps/genetics ; Humans ; *Hypocreales/genetics ; *Moths ; RNA, Ribosomal, 16S/genetics ; Soil ; Zygote ; }, abstract = {Chinese cordyceps is a well-known fungus-larva complex with medicinal and economic importance. At present the occurrence of Chinese cordyceps has not been fully illuminated. In this study, the microbial diversities of fertilized Thitarodes eggs from sites A (high occurrence rates of Chinese cordyceps), B (low occurrence rates), and C (no Chinese cordyceps) were analyzed using 16S rRNA and ITS gene-sequencing technique. The previous sequencing data of soil from the same sites were conjointly analyzed. The results showed that bacterial communities among the eggs were significantly different. The bacterial diversity and evenness were much higher on site A. Wolbachia was overwhelmingly predominant in the eggs of sites B and C, while Spiroplasma showed preference on site A. The fungal between-group differences in the eggs were not as significant as that of bacteria. Purpureocillium in Cordyceps-related families showed preference on site A. Wolbachia, Spiroplasma, and Purpureocillium were inferred to be closely related to Chinese cordyceps occurrence. Intra-kingdom and inter-kingdom network analyses suggest that closer correlations of microbial communities (especially closer fungal positive correlations) in fertilized eggs might promote Chinese cordyceps occurrence. Besides, metabolic pathway analysis showed that in fertilized eggs or soil the number of bacterial metabolic pathways with significant differences in every comparison between two sites was greater than that of fungi. Collectively, this study provides novel information about the occurrence of Chinese cordyceps, contributing to the large-scale artificial cultivation of Chinese cordyceps.}, } @article {pmid35765190, year = {2022}, author = {Weiland, SO and Detcharoen, M and Schlick-Steiner, BC and Steiner, FM}, title = {Analyses of locomotion, wing morphology, and microbiome in Drosophila nigrosparsa after recovery from antibiotics.}, journal = {MicrobiologyOpen}, volume = {11}, number = {3}, pages = {e1291}, pmid = {35765190}, issn = {2045-8827}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Drosophila ; Locomotion ; *Microbiota ; Tetracycline/pharmacology ; *Wolbachia/genetics ; }, abstract = {Antibiotics, such as tetracycline, have been frequently used to cure arthropods of Wolbachia endosymbionts. After the symbionts have been removed, the hosts must recover for some generations from the side effects of the antibiotics. However, most studies do not assess the direct and indirect longer-term effects of antibiotics used to remove Wolbachia, which may question the exact contribution of this endosymbiont to the effects observed. Here, we used the fly Drosophila nigrosparsa treated or not with tetracycline for three generations followed by two generations of recovery to investigate the effects of this antibiotic on the fly locomotion, wing morphology, and the gut microbiome. We found that antibiotic treatment did not affect fly locomotion two generations after being treated with the antibiotic. In addition, gut-microbiome restoration was tested as a more efficient solution to reduce the potential side effects of tetracycline on the microbiome. There was no significant difference in alpha diversity between gut restoration and other treatments, but the abundance of some bacterial taxa differed significantly between the gut-restoration treatment and the control. We conclude that in D. nigrosparsa the recovery period of two generations after being treated with the antibiotic is sufficient for locomotion, and suggest a general assessment of direct and indirect effects of antibiotics after a particular recovery time.}, } @article {pmid35746601, year = {2022}, author = {Soh, S and Ho, SH and Ong, J and Seah, A and Dickens, BS and Tan, KW and Koo, JR and Cook, AR and Sim, S and Tan, CH and Ng, LC and Lim, JT}, title = {Strategies to Mitigate Establishment under the Wolbachia Incompatible Insect Technique.}, journal = {Viruses}, volume = {14}, number = {6}, pages = {}, pmid = {35746601}, issn = {1999-4915}, mesh = {*Aedes/microbiology ; Animals ; Female ; Male ; Mosquito Control/methods ; Mosquito Vectors/microbiology ; Population Dynamics ; *Wolbachia ; }, abstract = {The Incompatible Insect Technique (IIT) strategy involves the release of male mosquitoes infected with the bacterium Wolbachia. Regular releases of male Wolbachia-infected mosquitoes can lead to the suppression of mosquito populations, thereby reducing the risk of transmission of vector-borne diseases such as dengue. However, due to imperfect sex-sorting under IIT, fertile Wolbachia-infected female mosquitoes may potentially be unintentionally released into the environment, which may result in replacement and failure to suppress the mosquito populations. As such, mitigating Wolbachia establishment requires a combination of IIT with other strategies. We introduced a simple compartmental model to simulate ex-ante mosquito population dynamics subjected to a Wolbachia-IIT programme. In silico, we explored the risk of replacement, and strategies that could mitigate the establishment of the released Wolbachia strain in the mosquito population. Our results suggest that mitigation may be achieved through the application of a sterile insect technique. Our simulations indicate that these interventions do not override the intended wild type suppression of the IIT approach. These findings will inform policy makers of possible ways to mitigate the potential establishment of Wolbachia using the IIT population control strategy.}, } @article {pmid35745561, year = {2022}, author = {Beld, L and Jung, H and Bulman, CA and Rosa, BA and Fischer, PU and Janetka, JW and Lustigman, S and Sakanari, JA and Mitreva, M}, title = {Aspartyl Protease Inhibitors as Anti-Filarial Drugs.}, journal = {Pathogens (Basel, Switzerland)}, volume = {11}, number = {6}, pages = {}, pmid = {35745561}, issn = {2076-0817}, support = {R01 AI159450/AI/NIAID NIH HHS/United States ; 1R01AI159450//National Institute of Allergy and Infectious Diseases/ ; }, abstract = {The current treatments for lymphatic filariasis and onchocerciasis do not effectively kill the adult parasitic nematodes, allowing these chronic and debilitating diseases to persist in millions of people. Thus, the discovery of new drugs with macrofilaricidal potential to treat these filarial diseases is critical. To facilitate this need, we first investigated the effects of three aspartyl protease inhibitors (APIs) that are FDA-approved as HIV antiretroviral drugs on the adult filarial nematode, Brugia malayi and the endosymbiotic bacteria, Wolbachia. From the three hits, nelfinavir had the best potency with an IC50 value of 7.78 µM, followed by ritonavir and lopinavir with IC50 values of 14.3 µM and 16.9 µM, respectively. The three APIs have a direct effect on killing adult B. malayi after 6 days of exposure in vitro and did not affect the Wolbachia titers. Sequence conservation and stage-specific gene expression analysis identified Bm8660 as the most likely primary aspartic protease target for these drug(s). Immunolocalization using antibodies raised against the Bm8660 ortholog of Onchocerca volvulus showed it is strongly expressed in female B. malayi, especially in metabolically active tissues such as lateral and dorsal/ventral chords, hypodermis, and uterus tissue. Global transcriptional response analysis using adult female B. pahangi treated with APIs identified four additional aspartic proteases differentially regulated by the three effective drugs, as well as significant enrichment of various pathways including ubiquitin mediated proteolysis, protein kinases, and MAPK/AMPK/FoxO signaling. In vitro testing against the adult gastro-intestinal nematode Trichuris muris suggested broad-spectrum potential for these APIs. This study suggests that APIs may serve as new leads to be further explored for drug discovery to treat parasitic nematode infections.}, } @article {pmid35737833, year = {2022}, author = {Morrison, AC and Reiner, RC and Elson, WH and Astete, H and Guevara, C and Del Aguila, C and Bazan, I and Siles, C and Barrera, P and Kawiecki, AB and Barker, CM and Vasquez, GM and Escobedo-Vargas, K and Flores-Mendoza, C and Huaman, AA and Leguia, M and Silva, ME and Jenkins, SA and Campbell, WR and Abente, EJ and Hontz, RD and Paz-Soldan, VA and Grieco, JP and Lobo, NF and Scott, TW and Achee, NL}, title = {Efficacy of a spatial repellent for control of Aedes-borne virus transmission: A cluster-randomized trial in Iquitos, Peru.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {119}, number = {26}, pages = {e2118283119}, pmid = {35737833}, issn = {1091-6490}, support = {P01 AI098670/AI/NIAID NIH HHS/United States ; }, mesh = {Adult ; *Aedes ; Animals ; Dengue/epidemiology/prevention & control ; Humans ; *Insect Repellents ; *Mosquito Control/standards ; *Mosquito Vectors ; Peru/epidemiology ; *Vector Borne Diseases/epidemiology/prevention & control/transmission ; Zika Virus ; Zika Virus Infection ; }, abstract = {Over half the world's population is at risk for viruses transmitted by Aedes mosquitoes, such as dengue and Zika. The primary vector, Aedes aegypti, thrives in urban environments. Despite decades of effort, cases and geographic range of Aedes-borne viruses (ABVs) continue to expand. Rigorously proven vector control interventions that measure protective efficacy against ABV diseases are limited to Wolbachia in a single trial in Indonesia and do not include any chemical intervention. Spatial repellents, a new option for efficient deployment, are designed to decrease human exposure to ABVs by releasing active ingredients into the air that disrupt mosquito-human contact. A parallel, cluster-randomized controlled trial was conducted in Iquitos, Peru, to quantify the impact of a transfluthrin-based spatial repellent on human ABV infection. From 2,907 households across 26 clusters (13 per arm), 1,578 participants were assessed for seroconversion (primary endpoint) by survival analysis. Incidence of acute disease was calculated among 16,683 participants (secondary endpoint). Adult mosquito collections were conducted to compare Ae. aegypti abundance, blood-fed rate, and parity status through mixed-effect difference-in-difference analyses. The spatial repellent significantly reduced ABV infection by 34.1% (one-sided 95% CI lower limit, 6.9%; one-sided P value = 0.0236, z = 1.98). Aedes aegypti abundance and blood-fed rates were significantly reduced by 28.6 (95% CI 24.1%, ∞); z = -9.11) and 12.4% (95% CI 4.2%, ∞); z = -2.43), respectively. Our trial provides conclusive statistical evidence from an appropriately powered, preplanned cluster-randomized controlled clinical trial of the impact of a chemical intervention, in this case a spatial repellent, to reduce the risk of ABV transmission compared to a placebo.}, } @article {pmid35735896, year = {2022}, author = {Nian, X and Tao, X and Xiao, Z and Wang, D and He, Y}, title = {Effects of Sublethal Concentrations of Tetracycline Hydrochloride on the Biological Characteristics and Wolbachia Titer in Parthenogenesis Trichogramma pretiosum.}, journal = {Insects}, volume = {13}, number = {6}, pages = {}, pmid = {35735896}, issn = {2075-4450}, support = {2021A1515010784//Natural Science Foundation of Guangdong/ ; 31772219//National Natural Science Foundation of China/ ; 2019KJ125//Guangdong Provincial Special Fund for Modern Agriculture Industry Technology Innovation Teams/ ; }, abstract = {Trichogramma pretiosum Riley is an important natural enemy and biological control agent of lepidopteran pests. Wolbachia is an intracellular endosymbiont that induces parthenogenesis in the parasitoid T. pretiosum. In this paper, the sublethal effects of the antibiotic tetracycline hydrochloride on the development and reproduction of T. pretiosum were studied. Emerged females were fed with sublethal concentrations (LC5, LC15, and LC35) of tetracycline for ten generations. The biological parameters (longevity, parasitized eggs, and fecundity) of treated females significantly reduced compared with the control Moreover, the percentage of female offspring in the treatments significantly reduced, but the percentage of male offspring significantly increased. In addition, the Wolbachia titer sharply reduced after two generations of antibiotic treatments, but it could still be detected even after ten successive generations of antibiotic treatments, which indicated that Wolbachia was not completely removed by sublethal concentrations of tetracycline. The control lines with higher Wolbachia titers produced more female offspring than the tetracycline treatments with lower Wolbachia titers, indicating that the Wolbachia titer affected the sex determination of T. pretiosum. Our results show that sublethal concentrations of tetracycline had adverse effects on the development of T. pretiosum, and Wolbachia titers affected the sexual development of T. pretiosum eggs.}, } @article {pmid35735886, year = {2022}, author = {Yan, ZC and Qi, GY and Yao, TY and Li, YX}, title = {Mitochondrial Genomes of Two Asexual Trichogramma (Hymenoptera: Trichogrammatidae) Strains and Comparison with Their Sexual Relatives.}, journal = {Insects}, volume = {13}, number = {6}, pages = {}, pmid = {35735886}, issn = {2075-4450}, support = {CX(21)3008//Jiangsu Agricultural Science and Technology Innovation Fund/ ; Grant No. 202030472//Shuangchuang Doctor Foundation of Jiangsu Province/ ; Grant No. 804018//Nanjing Agricultural University/ ; }, abstract = {Despite its substantial costs, sexual reproduction dominates in animals. One popular explanation for the paradox of sex is that asexual reproduction is more likely to accumulate deleterious mutations than sexual reproduction. To test this hypothesis, we compared the mitogenomes of two asexual wasp strains, Trichogramma cacoeciae and T. pretiosum, to their sexual relatives. These two asexual strains represent two different transition mechanisms in Trichogramma from sexual to asexual reproduction. Asexual T. pretiosum is induced by Wolbachia, while T. cacoeciae presumably originated from interspecific hybridization. We sequenced and assembled complete mitochondrial genomes of asexual T. cacoeciae and T. pretiosum. Compared to four sexual relatives, we found no evidence of higher mutation accumulation in asexual Trichogramma mitogenomes than in their sexual relatives. We also did not detect any relaxed selection in asexual Trichogramma mitogenomes. In contrast, the intensified selection was detected in Nad1 and Nad4 of the asexual T. pretiosum mitogenome, suggesting more purifying selection. In summary, no higher mitochondrial mutation accumulation was detected in these two asexual Trichogramma strains. This study provides a basis for further investigating mitochondrial evolution and asexual reproduction in Trichogramma.}, } @article {pmid35733096, year = {2022}, author = {da Silva, H and Oliveira, TMP and Sabino, EC and Alonso, DP and Sallum, MAM}, title = {Bacterial diversity in Haemagogus leucocelaenus (Diptera: Culicidae) from Vale do Ribeira, São Paulo, Brazil.}, journal = {BMC microbiology}, volume = {22}, number = {1}, pages = {161}, pmid = {35733096}, issn = {1471-2180}, mesh = {Animals ; Brazil ; *Culicidae ; Humans ; *Mercury ; Mosquito Vectors ; *Yellow Fever ; }, abstract = {INTRODUCTION: Mosquitoes (Diptera: Culicidae) are vectors that transmit numerous pathogens to humans and other vertebrates. Haemagogus leucocelaenus is a mosquito associated with transmission of yellow fever virus. The insect gut harbors a variety of microorganisms that can live and multiply within it, thus contributing to digestion, nutrition, and development of its host. The composition of bacterial communities in mosquitoes can be influenced by both biotic and abiotic factors. The goal of this study was to investigate the bacterial diversity of Hg. leucocelaenus and verify the differences between the bacterial communities in Hg. leucocelaenus from three different locations in the Atlantic tropical rain forest and southeastern state of São Paulo State, Brazil.

RESULTS: The phylum Proteobacteria was found in mosquitoes collected from the three selected study sites. More than 50% of the contigs belong to Wolbachia, followed by 5% Swaminathania, and 3% Acinetobacter. The genus Serratia was found in samples from two locations.

CONCLUSIONS: Wolbachia was reported for the first time in this species and may indicates that the vector competence of the populations of the species can vary along its geographical distribution area. The presence of Serratia might facilitate viral invasion caused by the disruption of the midgut barrier via action of the SmEnhancin protein, which digests the mucins present in the intestinal epithelium.}, } @article {pmid35730939, year = {2022}, author = {Jones, MW and Fricke, LC and Thorpe, CJ and Vander Esch, LO and Lindsey, ARI}, title = {Infection Dynamics of Cotransmitted Reproductive Symbionts Are Mediated by Sex, Tissue, and Development.}, journal = {Applied and environmental microbiology}, volume = {88}, number = {13}, pages = {e0052922}, pmid = {35730939}, issn = {1098-5336}, mesh = {Animals ; *Coinfection ; Drosophila/microbiology ; Insecta ; Reproduction ; Symbiosis ; *Wolbachia/physiology ; }, abstract = {One of the most prevalent intracellular infections on earth is with Wolbachia, a bacterium in the Rickettsiales that infects a range of insects, crustaceans, chelicerates, and nematodes. Wolbachia is maternally transmitted to offspring and has profound effects on the reproduction and physiology of its hosts, which can result in reproductive isolation, altered vectorial capacity, mitochondrial sweeps, and even host speciation. Some populations stably harbor multiple Wolbachia strains, which can further contribute to reproductive isolation and altered host physiology. However, almost nothing is known about the requirements for multiple intracellular microbes to be stably maintained across generations while they likely compete for space and resources. Here, we use a coinfection of two Wolbachia strains ("wHa" and "wNo") in Drosophila simulans to define the infection and transmission dynamics of an evolutionarily stable double infection. We find that a combination of sex, tissue, and host development contributes to the infection dynamics of the two microbes and that these infections exhibit a degree of niche partitioning across host tissues. wHa is present at a significantly higher titer than wNo in most tissues and developmental stages, but wNo is uniquely dominant in ovaries. Unexpectedly, the ratio of wHa to wNo in embryos does not reflect those observed in the ovaries, indicative of strain-specific transmission dynamics. Understanding how Wolbachia strains interact to establish and maintain stable infections has important implications for the development and effective implementation of Wolbachia-based vector biocontrol strategies, as well as more broadly defining how cooperation and conflict shape intracellular communities. IMPORTANCEWolbachia is a maternally transmitted intracellular bacterium that manipulates the reproduction and physiology of arthropods, resulting in drastic effects on the fitness, evolution, and even speciation of its hosts. Some hosts naturally harbor multiple strains of Wolbachia that are stably transmitted across generations, but almost nothing is known about the factors that limit or promote these coinfections, which can have profound effects on the host's biology and evolution and are under consideration as an insect-management tool. Here, we define the infection dynamics of a known stably transmitted double infection in Drosophila simulans with an eye toward understanding the patterns of infection that might facilitate compatibility between the two microbes. We find that a combination of sex, tissue, and development all contributes to infection dynamics of the coinfection.}, } @article {pmid35730490, year = {2022}, author = {Krome, AK and Becker, T and Kehraus, S and Schiefer, A and Gütschow, M and Chaverra-Muñoz, L and Hüttel, S and Jansen, R and Stadler, M and Ehrens, A and Pogorevc, D and Müller, R and Hübner, MP and Hesterkamp, T and Pfarr, K and Hoerauf, A and Wagner, KG and König, GM}, title = {Corallopyronin A: antimicrobial discovery to preclinical development.}, journal = {Natural product reports}, volume = {39}, number = {9}, pages = {1705-1720}, doi = {10.1039/d2np00012a}, pmid = {35730490}, issn = {1460-4752}, mesh = {Anti-Bacterial Agents/pharmacology ; *Anti-Infective Agents/pharmacology ; *Biological Products/pharmacology ; Humans ; Lactones ; Water ; }, abstract = {Covering: August 1984 up to January 2022Worldwide, increasing morbidity and mortality due to antibiotic-resistant microbial infections has been observed. Therefore, better prevention and control of infectious diseases, as well as appropriate use of approved antibacterial drugs are crucial. There is also an urgent need for the continuous development and supply of novel antibiotics. Thus, identifying new antibiotics and their further development is once again a priority of natural product research. The antibiotic corallopyronin A was discovered in the 1980s in the culture broth of the Myxobacterium Corallococcus coralloides and serves, in the context of this review, as a show case for the development of a naturally occurring antibiotic compound. The review demonstrates how a hard to obtain, barely water soluble and unstable compound such as corallopyronin A can be developed making use of sophisticated production and formulation approaches. Corallopyronin A is a bacterial DNA-dependent RNA polymerase inhibitor with a new target site and one of the few representatives of this class currently in preclinical development. Efficacy against Gram-positive and Gram-negative pathogens, e.g., Chlamydia trachomatis, Orientia tsutsugamushi, Staphylococcus aureus, and Wolbachia has been demonstrated. Due to its highly effective in vivo depletion of Wolbachia, which are essential endobacteria of most filarial nematode species, and its robust macrofilaricidal efficacy, corallopyronin A was selected as a preclinical candidate for the treatment of human filarial infections. This review highlights the discovery and production optimization approaches for corallopyronin A, as well as, recent preclinical efficacy results demonstrating a robust macrofilaricidal effect of the anti-Wolbachia candidate, and the solid formulation strategy which enhances the stability as well as the bioavailability of corallopyronin A.}, } @article {pmid35729601, year = {2022}, author = {Gunasekaran, K and Sadanandane, C and Panneer, D and Kumar, A and Rahi, M and Dinesh, S and Vijayakumar, B and Krishnaraja, M and Subbarao, SK and Jambulingam, P}, title = {Sensitivity of wMel and wAlbB Wolbachia infections in Aedes aegypti Puducherry (Indian) strains to heat stress during larval development.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {221}, pmid = {35729601}, issn = {1756-3305}, mesh = {*Aedes ; Animals ; Female ; Heat-Shock Response ; Larva ; Male ; Mosquito Vectors ; Water ; *Wolbachia ; }, abstract = {BACKGROUND: ICMR-Vector Control Research Centre, Puducherry, India, developed two colonies of Aedes aegypti infected with wMel and wAlbB Wolbacia strains called Ae. aegypti (Pud) lines for dengue control. The sensitivity of wMel and wAlbB strains in Ae. aegypti (Pud) lines to heat stress was studied.

METHODS: wMel and wAlbB infected and uninfected Ae. aegypti larvae (first to fourth instars) were reared in the laboratory to adults at 26 °C, 30 °C, 36 °C and 40 °C constant temperatures and also 26-30 °C, 26-36 °C and 26-40 °C diurnal cyclic temperatures. The adults were tested for Wolbachia infection. Experiments were also carried out rearing the larvae under simulated field conditions in summer (April and June) under sunlight using fully open and half open bowls and also under sunlight and natural shade.

RESULTS: At 36 °C and 40 °C constant temperatures, complete larval mortality was observed. At 30 °C and 26 °C, no larval mortality occurred, but Wolbachia density was relatively low in wMel infected males compared to control (maintained at 26 ± 1 °C). At diurnal cyclic temperature of 26-40 °C, Wolbachia density was reduced in males of both the (Pud) lines, but not in females. At 26-36 °C, reduction in Wolbachia density was observed in wMel males but not in wAlbB males. At 26-30 °C, no significant reduction in Wolbachia density was observed with wMel and wAlbB strains. In simulated field conditions (April), under sunlight, the daytime water temperature reached a maximum of 35.7 °C in both full and half open bowls. No larval mortality occurred. Wolbachia frequency and density was reduced in wMel-infected Ae. aegypti (Pud) males from both type of bowls and in females from full open bowls, and in wAlbB males from half open bowls. In June, rearing of larvae under sunlight, the first-instar larvae experienced a maximum daytime water temperature of > 38 °C that caused complete mortality. No larval mortality was observed in bowls kept under shade (< 32 °C).

CONCLUSIONS: Exposure of larvae to higher rearing temperatures in the laboratory and simulated-field conditions reduced the densities of wMel and wAlbB strains particularly in males, but the impact was more pronounced for wMel strain. The actual effect of heat stress on the stability of these two Wolbachia strains needs to be tested under natural field conditions.}, } @article {pmid35726500, year = {2022}, author = {Dzul-Rosado, K and Maldonado-Borges, JI and Puerto-Manzano, FI and Lammoglia-Villagómez, MA and Becker, I and Sánchez-Montes, S}, title = {First exploratory study of bacterial pathogens and endosymbionts in head lice from a Mayan community in southern Mexico.}, journal = {Zoonoses and public health}, volume = {69}, number = {6}, pages = {729-736}, doi = {10.1111/zph.12982}, pmid = {35726500}, issn = {1863-2378}, mesh = {*Acinetobacter/genetics ; Animals ; Bacteria/genetics ; DNA ; Humans ; *Lice Infestations/epidemiology/veterinary ; Mexico/epidemiology ; *Pediculus/genetics/microbiology ; Phylogeny ; }, abstract = {Lice represent one of the most neglected group of vectors worldwide, particularly in Latin America. Records of bacterial agents related to head lice are non-existent in this region of the continent. Many of these communities often do not have adequate access to public services and/or health protection. The normalization of this condition prevents them from manifesting discomfort, such as bites and itching, which further aggravates the situation, as they can be vectors of important diseases. For this reason, the aim of this work was to identify the richness of bacterial pathogens (Acinetobacter, Bartonella, and Rickettsia) and endosymbionts (Wolbachia) in head lice of paediatric patients from the indigenous municipality of Hoctun, Yucatan, Mexico. DNA extraction was performed using the QIAamp DNA Mini Kit. For the detection of bacterial pathogens, fragments of the gltA, rpoB, and 16S rDNA genes were amplified. For the detection of Wolbachia, the wsp gene was amplified. Of the 28 lice analysed, the presence of two genera of bacterial pathogens was detected Acinetobacter (42.9% = 12/28) and Bartonella (7.14% = 2/28). We also detected the endosymbiont Wolbachia (71.42% = 20/28). Our results showed that DNA from three bacteria species (Acinetobacter baumannii, Bartonella quintana, and Wolbachia pipientis) was present with frequencies ranging from 3.57% to 71.42%. This work represents the first exploratory study of the diversity of agents associated with head lice (Pediculus humanus capitis) in Mexico and Latin America. Due to the findings generated in the present study, it is important to perform surveillance of head lice populations to identify the degree of spread of these pathogens and their impact on populations in the region.}, } @article {pmid35724982, year = {2023}, author = {Faulk, C}, title = {De novo sequencing, diploid assembly, and annotation of the black carpenter ant, Camponotus pennsylvanicus, and its symbionts by one person for $1000, using nanopore sequencing.}, journal = {Nucleic acids research}, volume = {51}, number = {1}, pages = {17-28}, pmid = {35724982}, issn = {1362-4962}, mesh = {Animals ; Humans ; *Ants/genetics/microbiology ; Diploidy ; Genome, Mitochondrial ; *High-Throughput Nucleotide Sequencing/economics/methods ; Nanopore Sequencing ; *Sequence Analysis, DNA/economics/methods ; Symbiosis ; Wolbachia/genetics/physiology ; Enterobacteriaceae/classification/genetics/physiology ; *Genome, Insect ; }, abstract = {The black carpenter ant (Camponotus pennsylvanicus) is a pest species found widely throughout North America. From a single individual I used long-read nanopore sequencing to assemble a phased diploid genome of 306 Mb and 60X coverage, with quality assessed by a 97.0% BUSCO score, improving upon other ant assemblies. The mitochondrial genome reveals minor rearrangements from other ants. The reads also allowed assembly of parasitic and symbiont genomes. I include a complete Wolbachia bacterial assembly with a size of 1.2 Mb, as well as a commensal symbiont Blochmannia pennsylvanicus, at 791 kb. DNA methylation and hydroxymethylation were measured at base-pair resolution level from the same reads and confirmed extremely low levels seen in the Formicidae family. There was moderate heterozygosity, with 0.16% of bases being biallelic from the parental haplotypes. Protein prediction yielded 14 415 amino acid sequences with 95.8% BUSCO score and 86% matching to previously known proteins. All assemblies were derived from a single MinION flow cell generating 20 Gb of sequence for a cost of $1047 including consumable reagents. Adding fixed costs for equipment brings the total for an ant-sized genome to less than $5000. All analyses were performed in 1 week on a single desktop computer.}, } @article {pmid35723456, year = {2022}, author = {Chen, L and Xiao, Q and Shi, M and Cheng, J and Wu, J}, title = {Detecting Wolbachia Strain wAlbB in Aedes albopictus Cell Lines.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {184}, pages = {}, doi = {10.3791/63662}, pmid = {35723456}, issn = {1940-087X}, mesh = {*Aedes ; Animals ; Cell Line ; Mosquito Vectors ; *RNA Viruses ; *Wolbachia/physiology ; }, abstract = {As a maternally harbored endosymbiont, Wolbachia infects large proportions of insect populations. Studies have recently reported the successful regulation of RNA virus transmission using Wolbachia-transfected mosquitoes. Key strategies to control viruses include the manipulation of host reproduction via cytoplasmic incompatibility and the inhibition of viral transcripts via immune priming and competition for host-derived resources. However, the underlying mechanisms of the responses of Wolbachia-transfected mosquitoes to viral infection are poorly understood. This paper presents a protocol for the in vitro identification of Wolbachia infection at the nucleic acid and protein levels in Aedes albopictus (Diptera: Culicidae) Aa23 cells to enhance the understanding of the interactions between Wolbachia and its insect vectors. Through the combined use of polymerase chain reaction (PCR), quantitative PCR, western blot, and immunological analytical methods, a standard morphologic protocol has been described for the detection of Wolbachia-infected cells that is more accurate than the use of a single method. This approach may also be applied to the detection of Wolbachia infection in other insect taxa.}, } @article {pmid35715692, year = {2022}, author = {Hickin, ML and Kakumanu, ML and Schal, C}, title = {Effects of Wolbachia elimination and B-vitamin supplementation on bed bug development and reproduction.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {10270}, pmid = {35715692}, issn = {2045-2322}, mesh = {Animals ; *Bedbugs ; Dietary Supplements ; Female ; Nymph ; Reproduction ; *Vitamin B Complex/pharmacology ; *Wolbachia ; }, abstract = {Obligate blood feeders, such as Cimex lectularius (common bed bug), have symbiotic associations with nutritional endosymbionts that produce B-vitamins. To quantify the symbiont's contribution to host fitness in these obligate mutualisms, the symbiont must be eliminated and its absence rigorously confirmed. We developed and validated procedures for complete elimination of Wolbachia (Wb) in bed bugs and quantified development and reproduction in bed bugs with and without Wb and with and without B-vitamins supplementation. Aposymbiotic bed bugs had slower nymphal development, reduced adult survivorship, smaller adult size, fewer eggs per female, and lower hatch rate than bed bugs that harbored Wb. In aposymbiotic bed bugs that were fed B-vitamins-supplemented blood, nymph development time, adult survivorship and hatch rate recovered to control levels, but adult size and egg number only partially recovered. These results underscore the nutritional dependence of bed bugs on their Wb symbiont and suggest that Wb may provide additional nutritional benefits beyond the B-vitamin mix that we investigated.}, } @article {pmid35708345, year = {2022}, author = {Ettinger, CL and Byrne, FJ and Redak, RA and Stajich, JE}, title = {Metagenome-Assembled Genomes of Bacterial Symbionts Associated with Insecticide-Resistant and -Susceptible Individuals of the Glassy-Winged Sharpshooter (Homalodisca vitripennis).}, journal = {Microbiology resource announcements}, volume = {11}, number = {7}, pages = {e0050622}, pmid = {35708345}, issn = {2576-098X}, support = {14-0379-000-SA-2//Pierce's Disease Control Program/ ; 01170-002//California Department of Food and Agriculture (CDFA)/ ; 007011-003//California Department of Food and Agriculture (CDFA)/ ; //Canadian Institute for Advanced Research (ICRA)/ ; CA-R-PPA-5062-H//U.S. Department of Agriculture (USDA)/ ; 012604-002//USDA | Animal and Plant Health Inspection Service (APHIS)/ ; }, abstract = {The role of microbes in insecticide resistance is an emerging question. Here, we describe six metagenome-assembled genomes (MAGs) associated with the glassy-winged sharpshooter (Homalodisca vitripennis [Germar, 1821]) (Hemiptera, Cicadellidae). MAGs representing the obligate symbionts Candidatus Sulcia muelleri and Candidatus Baumannia cicadellinicola and the facultative symbiont Wolbachia were obtained from imidacloprid-resistant and imidacloprid-susceptible sharpshooters.}, } @article {pmid35701454, year = {2022}, author = {Dufault, SM and Tanamas, SK and Indriani, C and Utarini, A and Ahmad, RA and Jewell, NP and Simmons, CP and Anders, KL}, title = {Disruption of spatiotemporal clustering in dengue cases by wMel Wolbachia in Yogyakarta, Indonesia.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {9890}, pmid = {35701454}, issn = {2045-2322}, support = {R01 AI148127/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; R01AI148127/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes ; Animals ; Cluster Analysis ; *Dengue ; *Dengue Virus/genetics ; Humans ; Indonesia/epidemiology ; Pest Control, Biological ; *Wolbachia/genetics ; }, abstract = {Dengue exhibits focal clustering in households and neighborhoods, driven by local mosquito population dynamics, human population immunity, and fine scale human and mosquito movement. We tested the hypothesis that spatiotemporal clustering of homotypic dengue cases is disrupted by introduction of the arbovirus-blocking bacterium Wolbachia (wMel-strain) into the Aedes aegypti mosquito population. We analysed 318 serotyped and geolocated dengue cases (and 5921 test-negative controls) from a randomized controlled trial in Yogyakarta, Indonesia of wMel deployments. We find evidence of spatial clustering up to 300 m among the 265 dengue cases (3083 controls) in the untreated trial arm. Participant pairs enrolled within 30 days and 50 m had a 4.7-fold increase (compared to 95% CI on permutation-based null distribution: 0.1, 1.2) in the odds of being homotypic (i.e. potentially transmission-related) as compared to pairs occurring at any distance. In contrast, we find no evidence of spatiotemporal clustering among the 53 dengue cases (2838 controls) resident in the wMel-treated arm. Introgression of wMel Wolbachia into Aedes aegypti mosquito populations interrupts focal dengue virus transmission leading to reduced case incidence; the true intervention effect may be greater than the 77% efficacy measured in the primary analysis of the Yogyakarta trial.}, } @article {pmid35699129, year = {2022}, author = {Mulenga, GM and Namangala, B and Gummow, B}, title = {Prevalence of trypanosomes and selected symbionts in tsetse species of eastern Zambia.}, journal = {Parasitology}, volume = {149}, number = {11}, pages = {1406-1410}, pmid = {35699129}, issn = {1469-8161}, mesh = {Animals ; Enterobacteriaceae/genetics ; Insect Vectors/parasitology ; Prevalence ; *Trypanosoma/genetics ; *Trypanosomiasis, African/parasitology ; *Tsetse Flies/parasitology ; *Wolbachia/genetics ; Zambia/epidemiology ; }, abstract = {Insect symbionts have attracted attention for their potential use as anti-parasitic gene products in arthropod disease vectors. While tsetse species of the Luangwa valley have been extensively studied, less is known about the prevalence of symbionts and their interactions with the trypanosome parasite. Polymerase chain reaction was used to investigate the presence of Wolbachia and Sodalis bacteria, in tsetse flies infected with trypanosomes (Trypanosoma vivax, Trypanosoma congolense and Trypanosoma brucei). Out of 278 captured tsetse flies in eastern Zambia, 95.3% (n = 265, 95% CI = 92.8–97.8) carried endosymbionts: Wolbachia (79.1%, 95% CI 73.9–83.8) and Sodalis (86.3%, 95% CI 81.7–90.1). Overall, trypanosome prevalence was 25.5% (n = 71, 95% CI = 20.4–30.7), 10.8% (n = 30, 95% CI 7.1–14.4) for T. brucei, 1.4% (n = 4, 95% CI = 0.4–3.6) for both T. congolense and T. vivax, and 0.7% (n = 2, 95% CI 0.1–2.6) for T. b. rhodesiense. Out of 240 tsetse flies that were infected with Sodalis, trypanosome infection was reported in 40 tsetse flies (16.7%, 95% CI = 12.0–21.4) while 37 (16.8%, 95% CI 11.9–21.8) of the 220 Wolbachia infected tsetse flies were infected with trypanosomes. There was 1.3 times likelihood of T. brucei infection to be present when Wolbachia was present and 1.7 likelihood of T. brucei infection when Sodalis was present. Overall findings suggest absence of correlation between the presence of tsetse endosymbionts and tsetse with trypanosome infection. Lastly, the presence of pathogenic trypanosomes in tsetse species examined provided insights into the risk communities face, and the importance of African trypanosomiasis in the area.}, } @article {pmid35687421, year = {2022}, author = {Vörös, K and Becker, Z and Kónya, R and Arany-Tóth, A and Farkas, R}, title = {Application of Moxidectin and Ultrasound-Aided Injection of Melarsomine During the American Heartworm Society Recommended Treatment Protocol in Dirofilaria immitis Infected Dogs.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {22}, number = {7}, pages = {382-390}, doi = {10.1089/vbz.2021.0108}, pmid = {35687421}, issn = {1557-7759}, mesh = {Animals ; Arsenicals ; Clinical Protocols ; Dexamethasone/pharmacology/therapeutic use ; *Dirofilaria immitis ; *Dirofilariasis/diagnosis/drug therapy ; *Dog Diseases/drug therapy ; Dogs ; Doxycycline/therapeutic use ; *Filaricides/therapeutic use ; Macrolides/pharmacology/therapeutic use ; Triazines ; }, abstract = {The American Heartworm Society (AHS) recommends the three-dose alternate melarsomine therapeutic regimen, together with a macrocyclic lactone (ML) to reduce new infections and eliminate susceptible larvae and doxycycline against Wolbachia bacteria. Till now, only reports on ivermectin as an ML exist in the frame of this protocol. Between 2014 and 2020, the AHS protocol was used in 44 heartworm-positive dogs. Microfilaremic dogs were pretreated with prednisolone and clopidogrel for 1 week before the first moxidectin application. Moxidectin was applied on the 1st, 30th, 60th, and 90th therapeutic days. On the first day, dexamethasone and chloropyramine were used to avoid potential adverse effects caused by the destroyed microfilariae. During the 1st-28th days, doxycycline 10 mg/kg BID was given with probiotics. Adult heartworms were destroyed with melarsomine on the 60th, 90[th], and 91st days. Butorphanol and dexamethasone were given just before melarsomine injections. The depth of the intramuscular injection site was determined by ultrasound examination of the lumbar muscles. From the 60th day, dalteparin was applied for 10 days to decrease the chance of pulmonary thromboembolism. Moxidectin did not cause adverse reactions, even in microfilaremic dogs. Gastrointestinal side effects of doxycycline were observed in three (6%) dogs, they recovered after symptomatic therapy and by lowering the initial dose to 5 mg/kg BID. Transient anorexia and diarrhea were found in one (2%), and coughing and mild dyspnea in one (2%) animal as systemic post-therapeutic complications of melarsomine. No local side effects were observed in 13 (30%) dogs, mild local side effects occurred in 29 (66%) patients, and severe local swelling in 2 (4%) cases. All dogs recovered clinically by the 120th day and no microfilaremia was seen that time. An antigen test performed in 37/44 animals on the 271st day was also negative in all cases.}, } @article {pmid35686927, year = {2022}, author = {Kamiyama, T and Shimada-Niwa, Y and Tanaka, H and Katayama, M and Kuwabara, T and Mori, H and Kunihisa, A and Itoh, T and Toyoda, A and Niwa, R}, title = {Whole-genome sequencing analysis and protocol for RNA interference of the endoparasitoid wasp Asobara japonica.}, journal = {DNA research : an international journal for rapid publication of reports on genes and genomes}, volume = {29}, number = {4}, pages = {}, pmid = {35686927}, issn = {1756-1663}, support = {//JSPS KAKENHI/ ; //Japan Science and Technology Agency/ ; }, mesh = {Animals ; Drosophila/genetics ; Female ; Larva/parasitology ; Molecular Sequence Annotation ; RNA Interference ; RNA, Double-Stranded/genetics ; *Wasps/genetics ; }, abstract = {Asobara japonica is an endoparasitic wasp that parasitizes Drosophila flies. It synthesizes various toxic components in the venom gland and injects them into host larvae during oviposition. To identify and characterize these toxic components for enabling parasitism, we performed the whole-genome sequencing (WGS) and devised a protocol for RNA interference (RNAi) with A. japonica. Because it has a parthenogenetic lineage due to Wolbachia infection, we generated a clonal strain from a single wasp to obtain highly homogenous genomic DNA. The WGS analysis revealed that the estimated genome size was 322 Mb with a heterozygosity of 0.132%. We also performed RNA-seq analyses for gene annotation. Based on the qualified WGS platform, we cloned ebony-Aj, which encodes the enzyme N-β-alanyl dopamine synthetase, which is involved in melanin production. The microinjection of double-stranded RNA (dsRNA) targeting ebony-Aj led to body colour changes in adult wasps, phenocopying ebony-Dm mutants. Furthermore, we identified putative venom genes as a target of RNAi, confirming that dsRNA injection-based RNAi specifically suppressed the expression of the target gene in wasp adults. Taken together, our results provide a powerful genetic toolkit for studying the molecular mechanisms of parasitism.}, } @article {pmid35686658, year = {2022}, author = {Szklarzewicz, T and Kalandyk-Kołodziejczyk, M and Michalik, A}, title = {Ovary structure and symbiotic associates of a ground mealybug, Rhizoecus albidus (Hemiptera, Coccomorpha: Rhizoecidae) and their phylogenetic implications.}, journal = {Journal of anatomy}, volume = {241}, number = {3}, pages = {860-872}, pmid = {35686658}, issn = {1469-7580}, mesh = {Animals ; Female ; *Hemiptera/anatomy & histology/chemistry/microbiology ; In Situ Hybridization, Fluorescence ; Ovary/anatomy & histology ; Phylogeny ; Symbiosis ; }, abstract = {The ovary structure and the organization of its symbiotic system of the ground mealybug, Rhizoecus albidus (Rhizoecidae), were examined by means of microscopic and molecular methods. Each of the paired elongated ovaries of R. albidus is composed of circa one hundred short telotrophic-meroistic ovarioles, which are radially arranged along the distal part of the lateral oviduct. Analysis of serial sections revealed that each ovariole contains four germ cells: three trophocytes (nurse cells) occupying the tropharium and a single oocyte in the vitellarium. The ovaries are accompanied by giant cells termed bacteriocytes which are tightly packed with large pleomorphic bacteria. Their identity as Brownia rhizoecola (Bacteroidetes) was confirmed by means of amplicon sequencing and fluorescence in situ hybridization techniques. Moreover, to our knowledge, this is the first report on the morphology and ultrastructure of the Brownia rhizoecola bacterium. In the bacteriocyte cytoplasm bacteria Brownia co-reside with sporadic rod-shaped smaller bacteria, namely Wolbachia (Proteobacteria: Alphaproteobacteria). Both symbionts are transmitted to the next generation vertically (maternally), that is, via female germline cells. We documented that, at the time when ovarioles contain oocytes at the vitellogenic stage, these symbionts leave the bacteriocytes and move toward the neck region of ovarioles (i.e. the region between tropharium and vitellarium). Next, the bacteria enter the cytoplasm of follicular cells surrounding the basal part of the tropharium, leave them and enter the space between the follicular epithelium and surface of the nutritive cord connecting the tropharium and vitellarium. Finally, they gather in the deep depression of the oolemma at the anterior pole of the oocyte in the form of a 'symbiont ball'. Our results provide further arguments strongly supporting the validity of the recent changes in the classification of mealybugs, which involved excluding ground mealybugs from the Pseudococcidae family and raising them to the rank of their own family Rhizoecidae.}, } @article {pmid35678877, year = {2022}, author = {Araújo, NJS and Macêdo, MJF and de Morais, LP and da Cunha, FAB and de Matos, YMLS and de Almeida, RS and Braga, MFBM and Coutinho, HDM}, title = {Control of arboviruses vectors using biological control by Wolbachia pipientis: a short review.}, journal = {Archives of microbiology}, volume = {204}, number = {7}, pages = {376}, pmid = {35678877}, issn = {1432-072X}, mesh = {Animals ; *Arboviruses ; Insect Vectors/microbiology ; *Wolbachia ; }, abstract = {The number of arbovirus cases has increased in recent years, demonstrating a need for investing in effective control actions. Among these actions, are strategies using biological control vectors, a field where Wolbachia pipientis has shown itself as useful. Wolbachia pipientis, an obligatory intracellular Gram-negative bacteria, which parasites arthropods naturally or through laboratory-induced infections, is capable of manipulating the reproduction of its host. A systematic literature review gathering studies on this bacteria over last 10 years (2007-2021) was performed given its important role in the reduction of insect disease vectors. 111 articles were found, from which 78 were used in this study. Information on the Wolbachia biology, mechanism of action and potential for the biological control of insect disease vectors was gathered. The present study may contribute to the knowledge surrounding the bacterium, as well as stimulate the production of other studies with the same theme.}, } @article {pmid35678697, year = {2022}, author = {Withers, AJ and Rice, A and de Boer, J and Donkersley, P and Pearson, AJ and Chipabika, G and Karangwa, P and Uzayisenga, B and Mensah, BA and Mensah, SA and Nkunika, POY and Kachigamba, D and Smith, JA and Jones, CM and Wilson, K}, title = {The distribution of covert microbial natural enemies of a globally invasive crop pest, fall armyworm, in Africa: Enemy release and spillover events.}, journal = {The Journal of animal ecology}, volume = {91}, number = {9}, pages = {1826-1841}, pmid = {35678697}, issn = {1365-2656}, support = {BB/P023444/1//Global Challenges Research Fund/ ; NE/L002604/1//Natural Environment Research Council/ ; }, mesh = {Animals ; Introduced Species ; Kenya ; *Nucleopolyhedroviruses ; Spodoptera ; *Wolbachia ; }, abstract = {Invasive species pose a significant threat to biodiversity and agriculture world-wide. Natural enemies play an important part in controlling pest populations, yet we understand very little about the presence and prevalence of natural enemies during the early invasion stages. Microbial natural enemies of fall armyworm Spodoptera frugiperda are known in its native region, however, they have not yet been identified in Africa where fall armyworm has been an invasive crop pest since 2016. Larval samples were screened from Malawi, Rwanda, Kenya, Zambia, Sudan and Ghana for the presence of four different microbial natural enemies; two nucleopolyhedroviruses, Spodoptera frugiperda NPV (SfMNPV) and Spodoptera exempta NPV (SpexNPV); the fungal pathogen Metarhizium rileyi; and the bacterium Wolbachia. This study aimed to identify which microbial pathogens are present in invasive fall armyworm, and determine the geographical, meteorological and temporal variables that influence prevalence. Within 3 years of arrival, fall armyworm was exposed to all four microbial natural enemies. SfMNPV probably arrived with fall armyworm from the Americas, but this is the first putative evidence of host spillover from Spodoptera exempta (African armyworm) to fall armyworm for the endemic pathogen SpexNPV and for Wolbachia. It is also the first confirmed incidence of M. rileyi infecting fall armyworm in Africa. Natural enemies were localised, with variation being observed both nationally and temporally. The prevalence of SfMNPV (the most common natural enemy) was predominantly explained by variables associated with the weather; declining with increasing rainfall and increasing with temperature. However, virus prevalence also increased as the growing season progressed. The infection of an invasive species with a natural enemy from its native range and novel pathogens specific to its new range has important consequences for understanding the population ecology of invasive species and insect-pathogen interactions. Additionally, while it is widely known that temporal and geographic factors affect insect populations, this study reveals that these are important in understanding the distribution of microbial natural enemies associated with invasive pests during the early stages of invasion, and provide baseline data for future studies.}, } @article {pmid35671755, year = {2022}, author = {Tvedte, ES and Gasser, M and Zhao, X and Tallon, LJ and Sadzewicz, L and Bromley, RE and Chung, M and Mattick, J and Sparklin, BC and Dunning Hotopp, JC}, title = {Accumulation of endosymbiont genomes in an insect autosome followed by endosymbiont replacement.}, journal = {Current biology : CB}, volume = {32}, number = {12}, pages = {2786-2795.e5}, pmid = {35671755}, issn = {1879-0445}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Chromosomes ; Drosophila/genetics/microbiology ; Gene Transfer, Horizontal ; Genome ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Eukaryotic genomes can acquire bacterial DNA via lateral gene transfer (LGT).[1] A prominent source of LGT is Wolbachia,[2] a widespread endosymbiont of arthropods and nematodes that is transmitted maternally through female germline cells.[3,4] The DNA transfer from the Wolbachia endosymbiont wAna to Drosophila ananassae is extensive[5-7] and has been localized to chromosome 4, contributing to chromosome expansion in this lineage.[6] As has happened frequently with claims of bacteria-to-eukaryote LGT, the contribution of wAna transfers to the expanded size of D. ananassae chromosome 4 has been specifically contested[8] owing to an assembly where Wolbachia sequences were classified as contaminants and removed.[9] Here, long-read sequencing with DNA from a Wolbachia-cured line enabled assembly of 4.9 Mbp of nuclear Wolbachia transfers (nuwts) in D. ananassae and a 24-kbp nuclear mitochondrial transfer. The nuwts are <8,000 years old in at least two locations in chromosome 4 with at least one whole-genome integration followed by rapid extensive duplication of most of the genome with regions that have up to 10 copies. The genes in nuwts are accumulating small indels and mobile element insertions. Among the highly duplicated genes are cifA and cifB, two genes associated with Wolbachia-mediated Drosophila cytoplasmic incompatibility. The wAna strain that was the source of nuwts was subsequently replaced by a different wAna endosymbiont. Direct RNA Nanopore sequencing of Wolbachia-cured lines identified nuwt transcripts, including spliced transcripts, but functionality, if any, remains elusive.}, } @article {pmid35671324, year = {2022}, author = {Marriott, AE and Furlong Silva, J and Pionnier, N and Sjoberg, H and Archer, J and Steven, A and Kempf, D and Taylor, MJ and Turner, JD}, title = {A mouse infection model and long-term lymphatic endothelium co-culture system to evaluate drugs against adult Brugia malayi.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {6}, pages = {e0010474}, pmid = {35671324}, issn = {1935-2735}, support = {NC/M00175X/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; NC/S001131/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; }, mesh = {Animals ; *Brugia malayi ; Coculture Techniques ; Disease Models, Animal ; Endothelial Cells ; Endothelium, Lymphatic ; Female ; Interleukin-5 ; Male ; Mice ; Mice, Inbred BALB C ; *Wolbachia ; }, abstract = {The development of new drugs targeting adult-stage lymphatic filarial nematodes is hindered by the lack of a robust long-term in vitro culture model. Testing potential direct-acting and anti-Wolbachia therapeutic candidates against adult lymphatic filariae in vitro requires their propagation via chronic infection of gerbils. We evaluated Brugia malayi parasite burden data from male Mongolian gerbils compared with two immune-deficient mouse strains highly susceptible to B. malayi: CB.17 Severe-Combined Immmuno-Deficient (SCID) and interleukin-4 receptor alpha, interleukin-5 double knockout (IL-4Rα-/-IL-5-/-) mice. Adult worms generated in IL-4Rα-/-IL-5-/- mice were tested with different feeder cells (human embryonic kidney cells, human adult dermal lymphatic endothelial cells and human THP-1 monocyte differentiated macrophages) and comparative cell-free conditions to optimise and validate a long-term in vitro culture system. Cultured parasites were compared against those isolated from mice using motility scoring, metabolic viability assay (MTT), ex vivo microfilariae release assay and Wolbachia content by qPCR. A selected culture system was validated as a drug screen using reference anti-Wolbachia (doxycycline, ABBV-4083 / flubentylosin) or direct-acting compounds (flubendazole, suramin). BALB/c IL-4Rα-/-IL-5-/- or CB.17 SCID mice were superior to Mongolian gerbils in generating adult worms and supporting in vivo persistence for periods of up to 52 weeks. Adult females retrieved from BALB/c IL-4Rα-/-IL-5-/- mice could be cultured for up to 21 days in the presence of a lymphatic endothelial cell co-culture system with comparable motility, metabolic activity and Wolbachia titres to those maintained in vivo. Drug studies confirmed significant Wolbachia depletions or direct macrofilaricidal activities could be discerned when female B. malayi were cultured for 14 days. We therefore demonstrate a novel methodology to generate adult B. malayi in vivo and accurately evaluate drug efficacy ex vivo which may be adopted for drug screening with the dual benefit of reducing overall animal use and improving anti-filarial drug development.}, } @article {pmid35668540, year = {2022}, author = {Sarwar, MS and Jahan, N and Ali, A and Yousaf, HK and Munzoor, I}, title = {Establishment of Wolbachia infection in Aedes aegypti from Pakistan via embryonic microinjection and semi-field evaluation of general fitness of resultant mosquito population.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {191}, pmid = {35668540}, issn = {1756-3305}, mesh = {*Aedes/microbiology ; Animals ; *Dengue/prevention & control ; Female ; Male ; Microinjections ; Mosquito Vectors/microbiology ; Pakistan ; Pest Control, Biological ; *Wolbachia ; }, abstract = {BACKGROUND: Dengue is a mosquito-borne viral disease that is mainly spread by Aedes aegypti. It is prevalent on five continents, predominantly in tropical and sub-tropical zones across the world. Wolbachia bacteria have been extensively used in vector control strategies worldwide. The focus of the current study was to obtain a natural population of Ae. aegypti harbouring Wolbachia and to determine the impact of this bacteria on the new host in a semi-field environment.

METHODS: Wolbachia-infected Aedes albopictus was collected from the city of Lahore, Punjab, Pakistan, and Wolbachia were successfully introduced into laboratory-reared Ae. aegypti via embryonic microinjection. The stable vertical transmission of wAlbB in the host population was observed for eight generations, and the impact of Wolbachia on the general fitness of the host was evaluated in semi-field conditions.

RESULTS: In the laboratory and semi-field experiments, wAlbB Wolbachia presented a strong cytoplasmic incompatibility (CI) effect, evidenced as zero egg hatching, in crosses between Wolbachia-infected males and wild (uninfected) females of Ae. aegypti. Wolbachia infection had no noticeable impact on the general fitness (P > 0.05), fecundity, body size (females and males) and mating competitiveness of the new host, Ae. aegypti. However, there was a significant decrease in female fertility (egg hatch) (P < 0.001). In addition, under starvation conditions, there was a remarkable decrease (P < 0.0001) in the life span of Wolbachia-infected females compared to uninfected females (4 vs. > 5 days, respectively).

CONCLUSIONS: Wolbachia strain wAlbB has a great potential to control the dengue vector in Ae. aegypti populations by producing 100% CI with a limited burden on its host in natural field conditions. This strain can be used as a biological tool against vector-borne diseases.}, } @article {pmid35666732, year = {2022}, author = {Bordenstein, SR and Bordenstein, SR}, title = {Widespread phages of endosymbionts: Phage WO genomics and the proposed taxonomic classification of Symbioviridae.}, journal = {PLoS genetics}, volume = {18}, number = {6}, pages = {e1010227}, pmid = {35666732}, issn = {1553-7404}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Arthropods ; *Bacteriophages/genetics ; Eukaryota ; Genomics ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are the most common obligate, intracellular bacteria in animals. They exist worldwide in arthropod and nematode hosts in which they commonly act as reproductive parasites or mutualists, respectively. Bacteriophage WO, the largest of Wolbachia's mobile elements, includes reproductive parasitism genes, serves as a hotspot for genetic divergence and genomic rearrangement of the bacterial chromosome, and uniquely encodes a Eukaryotic Association Module with eukaryotic-like genes and an ensemble of putative host interaction genes. Despite WO's relevance to genome evolution, selfish genetics, and symbiotic applications, relatively little is known about its origin, host range, diversification, and taxonomic classification. Here we analyze the most comprehensive set of 150 Wolbachia and phage WO assemblies to provide a framework for discretely organizing and naming integrated phage WO genomes. We demonstrate that WO is principally in arthropod Wolbachia with relatives in diverse endosymbionts and metagenomes, organized into four variants related by gene synteny, often oriented opposite the putative origin of replication in the Wolbachia chromosome, and the large serine recombinase is an ideal typing tool to distinguish the four variants. We identify a novel, putative lytic cassette and WO's association with a conserved eleven gene island, termed Undecim Cluster, that is enriched with virulence-like genes. Finally, we evaluate WO-like Islands in the Wolbachia genome and discuss a new model in which Octomom, a notable WO-like Island, arose from a split with WO. Together, these findings establish the first comprehensive Linnaean taxonomic classification of endosymbiont phages, including non-Wolbachia phages from aquatic environments, that includes a new family and two new genera to capture the collective relatedness of these viruses.}, } @article {pmid35664841, year = {2022}, author = {Montoya-Alonso, JA and García Rodríguez, SN and Carretón, E and Rodríguez Escolar, I and Costa-Rodríguez, N and Matos, JI and Morchón, R}, title = {Seroprevalence of Feline Heartworm in Spain: Completing the Epidemiological Puzzle of a Neglected Disease in the Cat.}, journal = {Frontiers in veterinary science}, volume = {9}, number = {}, pages = {900371}, pmid = {35664841}, issn = {2297-1769}, abstract = {Feline heartworm is a vector-borne zoonotic disease caused by Dirofilaria immitis. It is a cosmopolitan disease that is continuously expanding. Spain is considered an endemic country; however, although there are many published studies in dogs, feline heartworm has been poorly studied in this country. Thus, the objective was to analyze the exposure to D. immitis throughout Spain to complete the epidemiological map in the feline species. For this, 6,588 feline serum samples were analyzed for the presence of D. immitis antigens and antibodies against D. immitis and Wolbachia. The results were analyzed according to sex, age, breed, habitat, origin (owned or shelter cats), presence of clinical signs, use of preventive, location and climatology. The results showed a prevalence of 0.5% and a seroprevalence of 9.4%. The highest antibody seroprevalences were reported in the Canary Islands and the Balearic Islands (19.2 and 16%, respectively), as well as in the autonomous communities located on the Mediterranean coast (9.2-11.2%). Seropositive cats were found in both indoor and outdoor cats, and from 6 months of age. Furthermore, only 5.8% of cats received regular prophylactic treatment. The results show that feline dirofilariasis is widely distributed throughout the national territory and corroborate that, where infected dogs are present, there are cats exposed to the parasite. It is necessary to implement efficient awareness and prophylaxis measures to control the incidence and expansion of feline heartworm in Spain.}, } @article {pmid35652239, year = {2022}, author = {Powell, JR}, title = {Modifying mosquitoes to suppress disease transmission: Is the long wait over?.}, journal = {Genetics}, volume = {221}, number = {3}, pages = {}, pmid = {35652239}, issn = {1943-2631}, mesh = {*Aedes/genetics ; Animals ; *Dengue/epidemiology/prevention & control ; *Dengue Virus ; Humans ; Mosquito Vectors/genetics ; *Wolbachia/genetics ; }, abstract = {For more than 50 years it has been a dream of medical entomologists and public health workers to control diseases like malaria and dengue fever by modifying, through genetics and other methods, the arthropods that transmit them to humans. A brief synopsis of the history of these efforts as applied to mosquitoes is presented; none proved to be effective in reducing disease prevalence. Only in the last few years have novel approaches been developed or proposed that indicate the long wait may be over. Three recent developments are particularly promising: CRISPR-Cas9 driven genetic modification, shifting naturally occurring allele frequencies, and microbe-based modifications. The last is the furthest along in implementation. Dengue fever incidence has been reduced between 40% and 96% in 4 different regions of the world where Wolbachia-infected Aedes aegypti have been established in the field. It is not yet clear how sustainable such control programs will prove to be, but there is good reason for optimism. In light of this, the time is ripe for reinvigorated research on vectors, especially genetics. Vector-borne diseases primarily affect under-developed countries and thus have not received the attention they deserve from wealthier countries with well-developed and funded biomedical research establishments.}, } @article {pmid35644356, year = {2022}, author = {Arguni, E and Indriani, C and Rahayu, A and Supriyati, E and Yohan, B and Hayati, RF and Wardana, S and Tantowijoyo, W and Anshari, MR and Rahayu, E and Rubangi, and Ahmad, RA and Utarini, A and Simmons, CP and Sasmono, RT}, title = {Dengue virus population genetics in Yogyakarta, Indonesia prior to city-wide Wolbachia deployment.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {102}, number = {}, pages = {105308}, doi = {10.1016/j.meegid.2022.105308}, pmid = {35644356}, issn = {1567-7257}, mesh = {Animals ; Cities ; *Culicidae ; *Dengue/epidemiology ; *Dengue Virus ; Genetics, Population ; Genotype ; Humans ; Indonesia/epidemiology ; Phylogeny ; Serogroup ; *Wolbachia/genetics ; }, abstract = {Dengue has been endemic in Yogyakarta, Indonesia for decades. Here, we report the dengue epidemiology, entomology, and virology in Yogyakarta in 2016-2017, prior to the commencement of the Applying Wolbachia to Eliminate Dengue (AWED) randomized trial. Dengue epidemiological data were compiled and blood samples from dengue-suspected patients were tested for dengue virus (DENV). Ae. aegypti mosquito samples were caught from the field using BG-Sentinel traps and tested for the presence of DENV infection. Sequencing of the DENV E gene was used to determine the phylogeny and genotypes of circulating DENV. Within the last decade, the 2016-2017 dengue incidence was considered very high. Among the 649 plasma samples collected between March 2016-February 2017; and 36,910 mosquito samples collected between December 2016-May 2017, a total of 197 and 38 samples were DENV-positive by qRT-PCR, respectively. All four DENV serotypes were detected, with DENV-3 (n = 88; 44.67%) and DENV-1 (n = 87; 44.16%) as the predominant serotype, followed by DENV-4 (n = 12; 6.09%) and DENV-2 (n = 10; 5.08%). The Yogyakarta DENV-1 isolates were classified into Genotype I and IV, while DENV-2, DENV-3, and DENV-4 isolates were classified into the Cosmopolitan genotype, Genotype I, and Genotype II, respectively. Yogyakarta DENV isolates were closely related to Indonesian strains from neighboring Javanese cities, consistent with the endemic circulation of DENV on this highly populous island. Our study provides comprehensive baseline information on the DENV population genetic characteristics in Yogyakarta, which are useful as baseline data for the AWED trial and the future DENV surveillance in the city in the presence of a Wolbachia-infected Ae. aegypti population.}, } @article {pmid35631057, year = {2022}, author = {Lee, E and Hien Nguyen, T and Yen Nguyen, T and Nam Vu, S and Duong Tran, N and Trung Nghia, L and Mai Vien, Q and Dong Nguyen, T and Kriiger Loterio, R and Iturbe-Ormaetxe, I and Flores, HA and O'Neill, SL and Anh Dang, D and Simmons, CP and Fraser, JE}, title = {Transient Introgression of Wolbachia into Aedes aegypti Populations Does Not Elicit an Antibody Response to Wolbachia Surface Protein in Community Members.}, journal = {Pathogens (Basel, Switzerland)}, volume = {11}, number = {5}, pages = {}, pmid = {35631057}, issn = {2076-0817}, support = {OPP1180815//Bill & Melinda Gates Foundation/ ; OPP1153619//Bill & Melinda Gates Foundation/ ; 102591/WT_/Wellcome Trust/United Kingdom ; N/A//Macquarie Group Foundation/ ; }, abstract = {Wolbachia is an endosymbiotic bacterium that can restrict the transmission of human pathogenic viruses by Aedes aegypti mosquitoes. Recent field trials have shown that dengue incidence is significantly reduced when Wolbachia is introgressed into the local Ae. aegypti population. Female Ae. aegypti are anautogenous and feed on human blood to produce viable eggs. Herein, we tested whether people who reside on Tri Nguyen Island (TNI), Vietnam developed antibodies to Wolbachia Surface Protein (WSP) following release of Wolbachia-infected Ae. aegypti, as a measure of exposure to Wolbachia. Paired blood samples were collected from 105 participants before and after mosquito releases and anti-WSP titres were measured by ELISA. We determined no change in anti-WSP titres after ~30 weeks of high levels of Wolbachia-Ae. aegypti on TNI. These data suggest that humans are not exposed to the major Wolbachia surface antigen, WSP, following introgression of Wolbachia-infected Ae. aegypti mosquitoes.}, } @article {pmid35630383, year = {2022}, author = {Csorba, AB and Fora, CG and Bálint, J and Felföldi, T and Szabó, A and Máthé, I and Loxdale, HD and Kentelky, E and Nyárádi, II and Balog, A}, title = {Endosymbiotic Bacterial Diversity of Corn Leaf Aphid, Rhopalosiphum maidis Fitch (Hemiptera: Aphididae) Associated with Maize Management Systems.}, journal = {Microorganisms}, volume = {10}, number = {5}, pages = {}, pmid = {35630383}, issn = {2076-2607}, abstract = {In this study, different maize fields cultivated under different management systems were sampled to test corn leaf aphid, Rhopalosiphum maidis, populations in terms of total and endosymbiotic bacterial diversity. Corn leaf aphid natural populations were collected from traditionally managed maize fields grown under high agricultural and natural landscape diversity as well as conventionally treated high-input agricultural fields grown in monoculture and with fertilizers use, hence with low natural landscape diversity. Total bacterial community assessment by DNA sequencing was performed using the Illumina MiSeq platform. In total, 365 bacterial genera were identified and 6 endosymbiont taxa. A high abundance of the primary endosymbiont Buchnera and secondary symbionts Serratia and Wolbachia were detected in all maize crops. Their frequency was found to be correlated with the maize management system used, probably with fertilizer input. Three other facultative endosymbionts ("Candidatus Hamiltonella", an uncultured Rickettsiales genus, and Spiroplasma) were also recorded at different frequencies under the two management regimes. Principal components analyses revealed that the relative contribution of the obligate and dominant symbiont Buchnera to the aphid endosymbiotic bacterial community was 72%, whereas for the managed system this was only 16.3%. When facultative symbionts alone were considered, the effect of management system revealed a DNA diversity of 23.3%.}, } @article {pmid35623398, year = {2022}, author = {Agarwal, A and Sarma, DK and Chaurasia, D and Maan, HS}, title = {Novel molecular approaches to combat vectors and vector-borne viruses: Special focus on RNA interference (RNAi) mechanisms.}, journal = {Acta tropica}, volume = {233}, number = {}, pages = {106539}, doi = {10.1016/j.actatropica.2022.106539}, pmid = {35623398}, issn = {1873-6254}, mesh = {*Aedes ; Animals ; Antiviral Agents ; *Chikungunya Fever ; Mosquito Vectors ; RNA Interference ; *Zika Virus/genetics ; *Zika Virus Infection ; }, abstract = {Vector-borne diseases, such as dengue, chikungunya, zika, yellow fever etc pose significant burden among the infectious diseases globally, especially in tropical and sub-tropical regions. Globalization, deforestation, urbanization, climate change, uncontrolled population growth, inadequate waste management and poor vector-management infrastructure have all contributed to the expansion of vector habitats and subsequent increase in vector-borne diseases throughout the world. Conventional vector control methods, such as use of insecticides, have significant negative environmental repercussions in addition to developing resistance in vectors. Till date, a very few vaccines or antiviral therapies have been approved for the treatment of vector borne diseases. In this review, we have discussed emerging molecular approaches like CRISPR (clustered regularly interspaced short palindromic repeats)/Cas-9, sterile insect technique (SIT), release of insects carrying a dominant lethal (RIDL), Wolbachia (virus transmission blocking) and RNA interference (RNAi) to combat vector and vector-borne viruses. Due to the extensive advancements in RNAi research, a special focus has been given on its types, biogenesis, mechanism of action, delivery and experimental studies evaluating their application as anti-mosquito and anti-viral agent. These technologies appear to be highly promising in terms of contributing to vector control and antiviral drug development, and hence can be used to reduce global vector and vector-borne disease burden.}, } @article {pmid35618926, year = {2022}, author = {Bouyer, J and Maiga, H and Vreysen, MJB}, title = {Assessing the efficiency of Verily's automated process for production and release of male Wolbachia-infected mosquitoes.}, journal = {Nature biotechnology}, volume = {40}, number = {10}, pages = {1441-1442}, pmid = {35618926}, issn = {1546-1696}, mesh = {*Aedes ; Animals ; Male ; Mosquito Control ; Mosquito Vectors ; *Wolbachia ; }, } @article {pmid35618925, year = {2022}, author = {Crawford, JE and Hopkins, KC and Buchman, A and Zha, T and Howell, P and Kakani, E and Ohm, JR and Snoad, N and Upson, L and Holeman, J and Massaro, P and Dobson, SL and Mulligan, FS and White, BJ}, title = {Reply to: Assessing the efficiency of Verily's automated process for production and release of male Wolbachia-infected mosquitoes.}, journal = {Nature biotechnology}, volume = {40}, number = {10}, pages = {1443-1446}, pmid = {35618925}, issn = {1546-1696}, mesh = {*Aedes ; Animals ; Male ; Mosquito Control ; *Wolbachia ; }, } @article {pmid35615502, year = {2022}, author = {Wang, L and Li, C and Wang, X and Wang, G and Shang, S and Dou, Z and Luo, Y}, title = {Gut Lignocellulose Activity and Microbiota in Asian Longhorned Beetle and Their Predicted Contribution to Larval Nutrition.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {899865}, pmid = {35615502}, issn = {1664-302X}, abstract = {Anoplophora glabripennis (Asian longhorned beetle) is a wood-boring pest that can inhabit a wide range of healthy deciduous host trees in native and invaded areas. The gut microbiota plays important roles in the acquisition of nutrients for the growth and development of A. glabripennis larvae. Herein, we investigated the larval gut structure and studied the lignocellulose activity and microbial communities of the larval gut following feeding on different host trees. The larval gut was divided into foregut, midgut, and hindgut, of which the midgut is the longest, forming a single loop under itself. Microbial community composition and lignocellulose activity in larval gut extracts were correlated with host tree species. A. glabripennis larvae fed on the preferred host (Populus gansuensis) had higher lignocellulose activity and microbial diversity than larvae reared on either a secondary host (Salix babylonica) or a resistant host (Populus alba var. pyramidalis). Wolbachia was the most dominant bacteria in the gut of larvae fed on S. babylonica and P. alba var. pyramidalis, while Enterococcus and Gibbsiella were the most dominant in larvae fed on P. gansuensis, followed by Wolbachia. The lignocellulose-degrading fungus Fusarium solani was dominant in the larval gut fed on different host trees. Functional predictions of microbial communities in the larval gut fed on different resistant host trees suggested that they all play a role in degrading lignocellulose, detoxification, and fixing nitrogen, which likely contribute to the ability of these larvae to thrive in a broad range of host tree species.}, } @article {pmid35613073, year = {2022}, author = {Harumoto, T and Fukatsu, T}, title = {Perplexing dynamics of Wolbachia proteins for cytoplasmic incompatibility.}, journal = {PLoS biology}, volume = {20}, number = {5}, pages = {e3001644}, pmid = {35613073}, issn = {1545-7885}, mesh = {Cytoplasm ; Cytosol ; Humans ; Male ; Prophages ; Spermatozoa ; *Wolbachia ; }, abstract = {The mechanism of symbiont-induced cytoplasmic incompatibility (CI) has been a long-standing mystery. A new study on Wolbachia's Cif proteins in PLOS Biology provides supportive evidence for the "Host modification model," although the alternative "Toxin-antidote model" is still in the running.}, } @article {pmid35610655, year = {2022}, author = {Waymire, E and Duddu, S and Yared, S and Getachew, D and Dengela, D and Bordenstein, SR and Balkew, M and Zohdy, S and Irish, SR and Carter, TE}, title = {Wolbachia 16S rRNA haplotypes detected in wild Anopheles stephensi in eastern Ethiopia.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {178}, pmid = {35610655}, issn = {1756-3305}, support = {R15 AI151766/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Anopheles/genetics ; Ethiopia/epidemiology ; Haplotypes ; Humans ; *Malaria ; *Malaria, Falciparum/epidemiology ; *Malaria, Vivax ; Mosquito Vectors/genetics ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: About two out of three Ethiopians are at risk of malaria, a disease caused by the parasites Plasmodium falciparum and Plasmodium vivax. Anopheles stephensi, an invasive vector typically found in South Asia and the Middle East, was recently found to be distributed across eastern and central Ethiopia and is capable of transmitting both P. falciparum and P. vivax. The detection of this vector in the Horn of Africa (HOA) coupled with widespread insecticide resistance requires that new methods of vector control be investigated in order to control the spread of malaria. Wolbachia, a naturally occurring endosymbiotic bacterium of mosquitoes, has been identified as a potential vector control tool that can be explored for the control of malaria transmission. Wolbachia could be used to control the mosquito population through suppression or potentially decrease malaria transmission through population replacement. However, the presence of Wolbachia in wild An. stephensi in eastern Ethiopia is unknown. This study aimed to identify the presence and diversity of Wolbachia in An. stephensi across eastern Ethiopia.

METHODS: DNA was extracted from An. stephensi collected from eastern Ethiopia in 2018 and screened for Wolbachia using a 16S targeted PCR assay, as well as multilocus strain typing (MLST) PCR assays. Haplotype and phylogenetic analysis of the sequenced 16S amplicons were conducted to compare with Wolbachia from countries across Africa and Asia.

RESULTS: Twenty out of the 184 mosquitoes screened were positive for Wolbachia, with multiple haplotypes detected. In addition, phylogenetic analysis revealed two superclades, representing Wolbachia supergroups A and B (bootstrap values of 81 and 72, respectively) with no significant grouping of geographic location or species. A subclade with a bootstrap value of 89 separates the Ethiopian haplotype 2 from other sequences in that superclade.

CONCLUSIONS: These findings provide the first evidence of natural Wolbachia populations in wild An. stephensi in the HOA. They also identify the need for further research to confirm the endosymbiotic relationship between Wolbachia and An. stephensi and to investigate its utility for malaria control in the HOA.}, } @article {pmid35609042, year = {2022}, author = {Kaur, R and Leigh, BA and Ritchie, IT and Bordenstein, SR}, title = {The Cif proteins from Wolbachia prophage WO modify sperm genome integrity to establish cytoplasmic incompatibility.}, journal = {PLoS biology}, volume = {20}, number = {5}, pages = {e3001584}, pmid = {35609042}, issn = {1545-7885}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/metabolism ; Drosophila melanogaster/genetics ; Female ; Male ; Prophages/genetics ; Protamines/metabolism ; Spermatozoa ; *Wolbachia ; }, abstract = {Inherited microorganisms can selfishly manipulate host reproduction to drive through populations. In Drosophila melanogaster, germline expression of the native Wolbachia prophage WO proteins CifA and CifB cause cytoplasmic incompatibility (CI) in which embryos from infected males and uninfected females suffer catastrophic mitotic defects and lethality; however, in infected females, CifA expression rescues the embryonic lethality and thus imparts a fitness advantage to the maternally transmitted Wolbachia. Despite widespread relevance to sex determination, evolution, and vector control, the mechanisms underlying when and how CI impairs male reproduction remain unknown and a topic of debate. Here, we use cytochemical, microscopic, and transgenic assays in D. melanogaster to demonstrate that CifA and CifB proteins of wMel localize to nuclear DNA throughout the process of spermatogenesis. Cif proteins cause abnormal histone retention in elongating spermatids and protamine deficiency in mature sperms that travel to the female reproductive tract with Cif proteins. Notably, protamine gene knockouts enhance wild-type CI. In ovaries, CifA localizes to germ cell nuclei and cytoplasm of early-stage egg chambers; however, Cifs are absent in late-stage oocytes and subsequently in fertilized embryos. Finally, CI and rescue are contingent upon a newly annotated CifA bipartite nuclear localization sequence. Together, our results strongly support the Host modification model of CI in which Cifs initially modify the paternal and maternal gametes to bestow CI-defining embryonic lethality and rescue.}, } @article {pmid35602266, year = {2021}, author = {Hien, NT and Anh, DD and Le, NH and Yen, NT and Phong, TV and Nam, VS and Duong, TN and Nguyen, NB and Huong, DTT and Hung, LQ and Trinh, CNT and Hoang, NV and Mai, VQ and Nghia, LT and Dong, NT and Tho, LH and Kutcher, S and Hurst, TP and Montgomery, JL and Woolfit, M and Rances, E and Kyrylos, P and L Anders, K and Nguyen, L and Brown-Kenyon, J and Caird, A and McLean, BJ and Iturbe-Ormaetxe, I and Ritchie, SA and O'Neill, SL and Ryan, PA}, title = {Environmental factors influence the local establishment of Wolbachia in Aedes aegypti mosquitoes in two small communities in central Vietnam.}, journal = {Gates open research}, volume = {5}, number = {}, pages = {147}, pmid = {35602266}, issn = {2572-4754}, abstract = {Background: The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and subsequently shown to reduce transmission of dengue and other pathogens, under both laboratory and field conditions. Here we describe the entomological outcomes of wMel Wolbachia mosquito releases in two small communities in Nha Trang City in central Vietnam. Methods: The wMel strain of Wolbachia was backcrossed into local Aedes aegypti genotype and mosquito releases were undertaken by community members or by staff. Field monitoring was undertaken to track Wolbachia establishment in local Ae. aegypti mosquito populations. Ecological studies were undertaken to assess relationships between environmental factors and the spatial and temporal variability in Wolbachia infection prevalence in mosquitoes. Results: Releases of wMel Wolbachia Ae. aegypti mosquitoes in two small communities in Nha Trang City resulted in the initial establishment of Wolbachia in the local Ae. aegypti mosquito populations, followed by seasonal fluctuations in Wolbachia prevalence. There was significant small-scale spatial heterogeneity in Wolbachia infection prevalence in the Tri Nguyen Village site, resulting in the loss of wMel Wolbachia infection in mosquitoes in north and center areas, despite Wolbachia prevalence remaining high in mosquitoes in the south area. In the second site, Vinh Luong Ward, Wolbachia has persisted at a high level in mosquitoes throughout this site despite similar seasonal fluctuations in wMel Wolbachia prevalence. Conclusion: Seasonal variation in Wolbachia infection prevalence in mosquitoes was associated with elevated temperature conditions, and was possibly due to imperfect maternal transmission of Wolbachia. Heterogeneity in Wolbachia infection prevalence was found throughout one site, and indicates additional factors may influence Wolbachia establishment.}, } @article {pmid35604942, year = {2022}, author = {Ørsted, M and Yashiro, E and Hoffmann, AA and Kristensen, TN}, title = {Population bottlenecks constrain host microbiome diversity and genetic variation impeding fitness.}, journal = {PLoS genetics}, volume = {18}, number = {5}, pages = {e1010206}, pmid = {35604942}, issn = {1553-7404}, mesh = {Animals ; Drosophila melanogaster/genetics ; *Genetic Fitness ; Genetic Variation ; *Microbiota/genetics ; RNA, Ribosomal, 16S/genetics ; }, abstract = {It is becoming increasingly clear that microbial symbionts influence key aspects of their host's fitness, and vice versa. This may fundamentally change our thinking about how microbes and hosts interact in influencing fitness and adaptation to changing environments. Here we explore how reductions in population size commonly experienced by threatened species influence microbiome diversity. Consequences of such reductions are normally interpreted in terms of a loss of genetic variation, increased inbreeding and associated inbreeding depression. However, fitness effects of population bottlenecks might also be mediated through microbiome diversity, such as through loss of functionally important microbes. Here we utilise 50 Drosophila melanogaster lines with different histories of population bottlenecks to explore these questions. The lines were phenotyped for egg-to-adult viability and their genomes sequenced to estimate genetic variation. The bacterial 16S rRNA gene was amplified in these lines to investigate microbial diversity. We found that 1) host population bottlenecks constrained microbiome richness and diversity, 2) core microbiomes of hosts with low genetic variation were constituted from subsets of microbiomes found in flies with higher genetic variation, 3) both microbiome diversity and host genetic variation contributed to host population fitness, 4) connectivity and robustness of bacterial networks was low in the inbred lines regardless of host genetic variation, 5) reduced microbial diversity was associated with weaker evolutionary responses of hosts in stressful environments, and 6) these effects were unrelated to Wolbachia density. These findings suggest that population bottlenecks reduce hologenomic variation (combined host and microbial genetic variation). Thus, while the current biodiversity crisis focuses on population sizes and genetic variation of eukaryotes, an additional focal point should be the microbial diversity carried by the eukaryotes, which in turn may influence host fitness and adaptability with consequences for the persistence of populations.}, } @article {pmid35602967, year = {2022}, author = {Pollmann, M and Moore, LD and Krimmer, E and D'Alvise, P and Hasselmann, M and Perlman, SJ and Ballinger, MJ and Steidle, JLM and Gottlieb, Y}, title = {Highly transmissible cytoplasmic incompatibility by the extracellular insect symbiont Spiroplasma.}, journal = {iScience}, volume = {25}, number = {5}, pages = {104335}, pmid = {35602967}, issn = {2589-0042}, abstract = {Cytoplasmic incompatibility (CI) is a form of reproductive manipulation caused by maternally inherited endosymbionts infecting arthropods, like Wolbachia, whereby matings between infected males and uninfected females produce few or no offspring. We report the discovery of a new CI symbiont, a strain of Spiroplasma causing CI in the parasitoid wasp Lariophagus distinguendus. Its extracellular occurrence enabled us to establish CI in uninfected adult insects by transferring Spiroplasma-infected hemolymph. We sequenced the CI-Spiroplasma genome and did not find any homologues of any of the cif genes discovered to cause CI in Wolbachia, suggesting independent evolution of CI. Instead, the genome contains other potential CI-causing candidate genes, such as homologues of high-mobility group (HMG) box proteins that are crucial in eukaryotic development but rare in bacterial genomes. Spiroplasma's extracellular nature and broad host range encompassing medically and agriculturally important arthropods make it a promising tool to study CI and its applications.}, } @article {pmid35602212, year = {2021}, author = {Cunha, A}, title = {Trojan mosquitoes control dengue.}, journal = {Communications medicine}, volume = {1}, number = {}, pages = {17}, pmid = {35602212}, issn = {2730-664X}, abstract = {Dengue virus is transmitted by Aedes aegypti mosquitoes and causes the disease known as dengue. In a trial published in The New England Journal of Medicine, Utarini and colleagues report that release of wolbachia-infected A. aegypti populations in a dengue endemic area reduces the number of symptomatic cases and of hospitalisations.}, } @article {pmid35598650, year = {2022}, author = {Louzada-Flores, VN and Kramer, L and Brianti, E and Napoli, E and Mendoza-Roldan, JA and Bezerra-Santos, MA and Latrofa, MS and Otranto, D}, title = {Treatment with doxycycline is associated with complete clearance of circulating Wolbachia DNA in Dirofilaria immitis-naturally infected dogs.}, journal = {Acta tropica}, volume = {232}, number = {}, pages = {106513}, doi = {10.1016/j.actatropica.2022.106513}, pmid = {35598650}, issn = {1873-6254}, mesh = {Animals ; *Cell-Free Nucleic Acids ; *Dirofilaria immitis/genetics ; *Dirofilariasis/drug therapy ; *Dog Diseases/parasitology ; Dogs ; Doxycycline/pharmacology/therapeutic use ; *Wolbachia/genetics ; }, abstract = {Bacteria of the genus Wolbachia are endosymbionts of parasitic filarial nematodes, including Dirofilaria immitis, and are a target for the treatment of canine heartworm disease. In the present study, 53 naturally-infected dogs were divided in three groups, based on their positivity to D. immitis by antigen and Knott tests, to assess the efficacy of doxycycline treatment in eliminating Wolbachia from circulating blood. At T0, dogs that scored positive to both tests (G1) or to antigen only (G2) were submitted to doxycycline (10 mg/kg BID PO) treatment and to 10% Imidacloprid + 2.5% Moxidectin (Advocate®), while those negative to both tests (G3) received only 10% Imidacloprid + 2.5% Moxidectin (Advocate®). All dogs were followed-up for one year, monthly treated with Advocate® and regularly monitored by antigen and Knott tests. During the whole period, all blood samples were screened for Wolbachia-D. immitis DNA load by quantitative real-time PCR (qPCR). At T0, 88.2% of the microfilariemic dogs were positive for Wolbachia DNA, while none of the dogs from G2 or G3 were positive. Wolbachia DNA was no longer detectable in dogs from G1 following 1 month of doxycycline treatment and microfilariae (mfs) were cleared at T2. All dogs from the G1 and G2 were negative for D. immitis antigen at 12 months. Results of this study suggest that successful elimination of mfs by doxycycline is associated with complete clearance of Wolbachia DNA in D. immitis-naturally infected dogs.}, } @article {pmid35581290, year = {2022}, author = {Gomes, TMFF and Wallau, GL and Loreto, ELS}, title = {Multiple long-range host shifts of major Wolbachia supergroups infecting arthropods.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {8131}, pmid = {35581290}, issn = {2045-2322}, mesh = {Animals ; *Arthropods/genetics/microbiology ; Female ; Insecta/microbiology ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a genus of intracellular bacterial endosymbionts found in 20-66% of all insect species and a range of other invertebrates. It is classified as a single species, Wolbachia pipientis, divided into supergroups A to U, with supergroups A and B infecting arthropods exclusively. Wolbachia is transmitted mainly via vertical transmission through female oocytes, but can also be transmitted across different taxa by host shift (HS): the direct transmission of Wolbachia cells between organisms without involving vertically transmitted gametic cells. To assess the HS contribution, we recovered 50 orthologous genes from over 1000 Wolbachia genomes, reconstructed their phylogeny and calculated gene similarity. Of 15 supergroup A Wolbachia lineages, 10 have similarities ranging from 95 to 99.9%, while their hosts' similarities are around 60 to 80%. For supergroup B, four out of eight lineages, which infect diverse and distantly-related organisms such as Acari, Hemiptera and Diptera, showed similarities from 93 to 97%. These results show that Wolbachia genomes have a much higher similarity when compared to their hosts' genes, which is a major indicator of HS. Our comparative genomic analysis suggests that, at least for supergroups A and B, HS is more frequent than expected, occurring even between distantly-related species.}, } @article {pmid35560029, year = {2022}, author = {Thayanukul, P and Lertanantawong, B and Sirawaraporn, W and Charasmongkolcharoen, S and Chaibun, T and Jittungdee, R and Kittayapong, P}, title = {Simple, sensitive, and cost-effective detection of wAlbB Wolbachia in Aedes mosquitoes, using loop mediated isothermal amplification combined with the electrochemical biosensing method.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {5}, pages = {e0009600}, pmid = {35560029}, issn = {1935-2735}, mesh = {*Aedes/genetics ; Animals ; *Arbovirus Infections ; Cost-Benefit Analysis ; Humans ; Molecular Diagnostic Techniques ; Mosquito Vectors ; Nucleic Acid Amplification Techniques ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Wolbachia is an endosymbiont bacterium generally found in about 40% of insects, including mosquitoes, but it is absent in Aedes aegypti which is an important vector of several arboviral diseases. The evidence that Wolbachia trans-infected Ae. aegypti mosquitoes lost their vectorial competence and became less capable of transmitting arboviruses to human hosts highlights the potential of using Wolbachia-based approaches for prevention and control of arboviral diseases. Recently, release of Wolbachia trans-infected Ae. aegypti has been deployed widely in many countries for the control of mosquito-borne viral diseases. Field surveillance and monitoring of Wolbachia presence in released mosquitoes is important for the success of these control programs. So far, a number of studies have reported the development of loop mediated isothermal amplification (LAMP) assays to detect Wolbachia in mosquitoes, but the methods still have some specificity and cost issues.

We describe here the development of a LAMP assay combined with the DNA strand displacement-based electrochemical sensor (BIOSENSOR) method to detect wAlbB Wolbachia in trans-infected Ae. aegypti. Our developed LAMP primers used a low-cost dye detecting system and 4 oligo nucleotide primers which can reduce the cost of analysis while the specificity is comparable to the previous methods. The detection capacity of our LAMP technique was 1.4 nM and the detection limit reduced to 2.2 fM when combined with the BIOSENSOR. Our study demonstrates that a BIOSENSOR can also be applied as a stand-alone method for detecting Wolbachia; and it showed high sensitivity when used with the crude DNA extracts of macerated mosquito samples without DNA purification.

CONCLUSIONS/SIGNIFICANCE: Our results suggest that both LAMP and BIOSENSOR, either used in combination or stand-alone, are robust and sensitive. The methods have good potential for routine detection of Wolbachia in mosquitoes during field surveillance and monitoring of Wolbachia-based release programs, especially in countries with limited resources.}, } @article {pmid35547116, year = {2022}, author = {Weyandt, N and Aghdam, SA and Brown, AMV}, title = {Discovery of Early-Branching Wolbachia Reveals Functional Enrichment on Horizontally Transferred Genes.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {867392}, pmid = {35547116}, issn = {1664-302X}, abstract = {Wolbachia is a widespread endosymbiont of insects and filarial nematodes that profoundly influences host biology. Wolbachia has also been reported in rhizosphere hosts, where its diversity and function remain poorly characterized. The discovery that plant-parasitic nematodes (PPNs) host Wolbachia strains with unknown roles is of interest evolutionarily, ecologically, and for agriculture as a potential target for developing new biological controls. The goal of this study was to screen communities for PPN endosymbionts and analyze genes and genomic patterns that might indicate their role. Genome assemblies revealed 1 out of 16 sampled sites had nematode communities hosting a Wolbachia strain, designated wTex, that has highly diverged as one of the early supergroup L strains. Genome features, gene repertoires, and absence of known genes for cytoplasmic incompatibility, riboflavin, biotin, and other biosynthetic functions placed wTex between mutualist C + D strains and reproductive parasite A + B strains. Functional terms enriched in group L included protoporphyrinogen IX, thiamine, lysine, fatty acid, and cellular amino acid biosynthesis, while dN/dS analysis suggested the strongest purifying selection on arginine and lysine metabolism, and vitamin B6, heme, and zinc ion binding, suggesting these as candidate roles in PPN Wolbachia. Higher dN/dS pathways between group L, wPni from aphids, wFol from springtails, and wCfeT from cat fleas suggested distinct functional changes characterizing these early Wolbachia host transitions. PPN Wolbachia had several putative horizontally transferred genes, including a lysine biosynthesis operon like that of the mitochondrial symbiont Midichloria, a spirochete-like thiamine synthesis operon shared only with wCfeT, an ATP/ADP carrier important in Rickettsia, and a eukaryote-like gene that may mediate plant systemic acquired resistance through the lysine-to-pipecolic acid system. The Discovery of group L-like variants from global rhizosphere databases suggests diverse PPN Wolbachia strains remain to be discovered. These findings support the hypothesis of plant-specialization as key to shaping early Wolbachia evolution and present new functional hypotheses, demonstrating promise for future genomics-based rhizosphere screens.}, } @article {pmid35539006, year = {2022}, author = {Ugbe, FA and Shallangwa, GA and Uzairu, A and Abdulkadir, I}, title = {Theoretical modeling and design of some pyrazolopyrimidine derivatives as Wolbachia inhibitors, targeting lymphatic filariasis and onchocerciasis.}, journal = {In silico pharmacology}, volume = {10}, number = {1}, pages = {8}, pmid = {35539006}, issn = {2193-9616}, abstract = {Lymphatic filariasis and onchocerciasis are two common filarial diseases caused by a group of parasitic nematodes called filarial worms, which play host to the bacteria organism Wolbachia. One good treatment approach seeks Wolbachia as drug target. Here, a QSAR study was conducted to investigate the anti-wolbachia activities (pEC50) of 52 pyrazolopyrimidine analogues, while using the built model to predict the pEC50 values of the newly designed analogues. Density Functional Theory was used for the structural optimization, while the model building was based on Genetic Function Algorithm approach. The built QSAR model was validated thus: R[2] = 0.8104, R[2] adj = 0.7629, Q[2] cv = 0.6981, R[2] test = 0.7501 and cRp[2] = 0.7476. The predicted pEC50 of all newly designed compounds were higher than that of the template (43). The new compounds were; observed to pass the drug-likeness criteria, uniformly distributed to the brain, and found to be non-mutagenic. Also, the new compounds and the reference drug (doxycycline), were docked onto Ovarian Tumor (OTU) deubiquitinase receptor (PDB ID: 6W9O) using iGEMDOCK tool. This protein is known to help Wolbachia subvert host ubiquitin signaling. The resulting binding scores of the newly designed compounds except A5 were higher than that of doxycycline, while the protein-ligand interactions were majorly characterized by Hydrogen-bonding and hydrophobic interaction types. Therefore, the newly designed molecules could be developed as potential drug candidates for the treatment of lymphatic filariasis and onchocerciasis.}, } @article {pmid35538383, year = {2022}, author = {Fallon, AM}, title = {Muramidase, nuclease, or hypothetical protein genes intervene between paired genes encoding DNA packaging terminase and portal proteins in Wolbachia phages and prophages.}, journal = {Virus genes}, volume = {58}, number = {4}, pages = {327-349}, pmid = {35538383}, issn = {1572-994X}, mesh = {*Bacteriophages/genetics ; DNA Packaging ; Endodeoxyribonucleases ; Muramidase/genetics ; Prophages/genetics ; *Wolbachia/genetics ; }, abstract = {Genomes of the obligate intracellular alpha proteobacterium Wolbachia pipientis often encode prophage-like regions, and in a few cases, purified particles have been recovered. Because the structure of a conserved WO phage genome has been difficult to establish, we examined paired terminase and portal genes in Wolbachia phages and prophages, relative to those encoded by the gene transfer agent RcGTA from the free-living alpha proteobacterium Rhodobacter capsulatus. Terminase and portal proteins from Wolbachia have higher similarity to orthologs encoded by RcGTA than to orthologs encoded by bacteriophage lambda. In lambdoid phages, these proteins play key roles in assembly of mature phage particles, while in less well-studied gene transfer agents, terminase and portal proteins package random fragments of bacterial DNA, which could confound elucidation of WO phage genomes. In WO phages and prophages, terminase genes followed by a short gpW gene may be separated from the downstream portal gene by open-reading frames encoding a GH_25 hydrolase/muramidase, a PD-(D/E)XK nuclease, a hypothetical protein and/or a RelE/ParE toxin-antitoxin module. These aspects of gene organization, coupled with evidence for a low, non-inducible yield of WO phages, and the small size of WO phage particles described in the literature raise the possibility that Wolbachia prophage regions participate in processes that extend beyond conventional bacteriophage lysogeny and lytic replication. These intervening genes, and their possible relation to functions associated with GTAs, may contribute to variability among WO phage genomes recovered from physical particles and impact the ability of WO phages to act as transducing agents.}, } @article {pmid35532932, year = {2022}, author = {Strunov, A and Lerch, S and Blanckenhorn, WU and Miller, WJ and Kapun, M}, title = {Complex effects of environment and Wolbachia infections on the life history of Drosophila melanogaster hosts.}, journal = {Journal of evolutionary biology}, volume = {35}, number = {6}, pages = {788-802}, pmid = {35532932}, issn = {1420-9101}, support = {P 32275/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Drosophila melanogaster/genetics ; Female ; Fertility ; Longevity ; Male ; Reproduction ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia bacteria are common endosymbionts of many arthropods found in gonads and various somatic tissues. They manipulate host reproduction to enhance their transmission and confer complex effects on fitness-related traits. Some of these effects can serve to increase the survival and transmission efficiency of Wolbachia in the host population. The Wolbachia-Drosophila melanogaster system represents a powerful model to study the evolutionary dynamics of host-microbe interactions and infections. Over the past decades, there has been a replacement of the ancestral wMelCS Wolbachia variant by the more recent wMel variant in worldwide D. melanogaster populations, but the reasons remain unknown. To investigate how environmental change and genetic variation of the symbiont affect host developmental and adult life-history traits, we compared effects of both Wolbachia variants and uninfected controls in wild-caught D. melanogaster strains at three developmental temperatures. While Wolbachia did not influence any developmental life-history traits, we found that both lifespan and fecundity of host females were increased without apparent fitness trade-offs. Interestingly, wMelCS-infected flies were more fecund than uninfected and wMel-infected flies. By contrast, males infected with wMel died sooner, indicating sex-specific effects of infection that are specific to the Wolbachia variant. Our study uncovered complex temperature-specific effects of Wolbachia infections, which suggests that symbiont-host interactions in nature are strongly dependent on the genotypes of both partners and the thermal environment.}, } @article {pmid35531293, year = {2022}, author = {Gao, S and Ren, YS and Su, CY and Zhu, DH}, title = {High Levels of Multiple Phage WO Infections and Its Evolutionary Dynamics Associated With Wolbachia-Infected Butterflies.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {865227}, pmid = {35531293}, issn = {1664-302X}, abstract = {Wolbachia is a maternally inherited bacterium that is widely distributed among arthropods, in which it manipulates the reproduction of its hosts. Phage WO is the only bacteriophage known to infect Wolbachia, and may provide benefit to its host or arthropods. We screened for the presence of phage WO in Wolbachia-infected butterfly species for the first time, to investigate their diversity and evolutionary dynamics. All Wolbachia-infected butterfly species, including members of the families Hesperiidae, Lycaenidae, Nymphalidae, Papilionidae, and Pieridae, were found to harbor phage WO. Interestingly, 84% of 19 butterfly species, which were infected with a single Wolbachia strain harbored high levels of multiple phage types (ranging from 3 to 17 types), another three species harbored one or two phage types. For Wolbachia strains (ST-41, ST-19, ST-125 and ST-374) shared among various butterfly species, their host insects all harbored multiple phage types, while two Wolbachia strains (ST-297 and ST-wPcau) were found to infect one butterfly species, whose insect hosts harbored a single phage type, suggesting that horizontal transfer of Wolbachia between insects increased the likelihood of exposure to phages, resulting in increased phage genetic diversity. Twelve horizontal transmission events of phage WO were found, which shared common phage WO types among different Wolbachia strains associated with butterflies. Most horizontal transfer events involved different Wolbachia supergroups (A and B). Horizontal acquisition of phage WO might also occur between eukaryotes without Wolbachia transfer. Furthermore, 22 putative recombination events were identified in 13 of 16 butterfly species which harbored multiple phage types. These results showed that horizontal transfer of Wolbachia caused it to be exposed to the phage gene pool, and that horizontal transmission of phage WO, as well as intragenic recombination were important dynamics for phage WO genome evolution, which effectively promoted the high level of phage WO diversity associated with butterflies.}, } @article {pmid35526745, year = {2022}, author = {Nascimento da Silva, J and Calixto Conceição, C and Cristina Ramos de Brito, G and Costa Santos, D and Martins da Silva, R and Arcanjo, A and Henrique Ferreira Sorgine, M and de Oliveira, PL and Andrade Moreira, L and da Silva Vaz, I and Logullo, C}, title = {Wolbachia pipientis modulates metabolism and immunity during Aedes fluviatilis oogenesis.}, journal = {Insect biochemistry and molecular biology}, volume = {146}, number = {}, pages = {103776}, doi = {10.1016/j.ibmb.2022.103776}, pmid = {35526745}, issn = {1879-0240}, mesh = {*Aedes/microbiology ; Animals ; Oogenesis ; Symbiosis/physiology ; *Wolbachia/physiology ; }, abstract = {Wolbachia pipientis is a maternally transmitted bacterium that mostly colonizes arthropods, including the mosquito Aedes fluviatilis, potentially affecting different aspects of host physiology. This intracellular bacterium prefers gonadal tissue cells, interfering with the reproductive cycle of insects, arachnids, crustaceans, and nematodes. Wolbachia's ability to modulate the host's reproduction is related to its success in prevalence and frequency. Infecting oocytes is essential for vertical propagation, ensuring its presence in the germline. The mosquito Ae. fluviatilis is a natural host for this bacterium and therefore represents an excellent experimental model in the effort to understand host-symbiont interactions and the mutual metabolic regulation. The aim of this study was to comparatively describe metabolic changes in naturally Wolbachia-infected and uninfected ovaries of Ae. fluviatilis during the vitellogenic period of oogenesis, thus increasing the knowledge about Wolbachia parasitic/symbiotic mechanisms.}, } @article {pmid35489839, year = {2022}, author = {Reyna-Lara, A and Soriano-Paños, D and Arias-Castro, JH and Martínez, HJ and Gómez-Gardeñes, J}, title = {A metapopulation approach to identify targets for Wolbachia-based dengue control.}, journal = {Chaos (Woodbury, N.Y.)}, volume = {32}, number = {4}, pages = {041105}, doi = {10.1063/5.0087435}, pmid = {35489839}, issn = {1089-7682}, mesh = {Aedes/*microbiology ; Animals ; Chikungunya Fever/epidemiology/*prevention & control/transmission ; Dengue/epidemiology/*prevention & control/transmission ; Humans ; Mosquito Control/economics ; Mosquito Vectors/*microbiology ; Wolbachia/growth & development/*physiology ; Zika Virus Infection/epidemiology/*prevention & control/transmission ; }, abstract = {Over the last decade, the release of Wolbachia-infected Aedes aegypti into the natural habitat of this mosquito species has become the most sustainable and long-lasting technique to prevent and control vector-borne diseases, such as dengue, zika, or chikungunya. However, the limited resources to generate such mosquitoes and their effective distribution in large areas dominated by the Aedes aegypti vector represent a challenge for policymakers. Here, we introduce a mathematical framework for the spread of dengue in which competition between wild and Wolbachia-infected mosquitoes, the cross-contagion patterns between humans and vectors, the heterogeneous distribution of the human population in different areas, and the mobility flows between them are combined. Our framework allows us to identify the most effective areas for the release of Wolbachia-infected mosquitoes to achieve a large decrease in the global dengue prevalence.}, } @article {pmid35488963, year = {2022}, author = {Wangkeeree, J and Suwanchaisri, K and Roddee, J and Hanboonsong, Y}, title = {Selective Elimination of Wolbachia from the Leafhopper Yamatotettix flavovittatus Matsumura.}, journal = {Current microbiology}, volume = {79}, number = {6}, pages = {173}, pmid = {35488963}, issn = {1432-0991}, support = {SciGR 7/2564//faculty of science and technology, thammasat university/ ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Hemiptera/microbiology ; Rifampin/pharmacology ; Tetracycline/pharmacology ; *Wolbachia ; }, abstract = {Wolbachia infections affect the reproductive system and various biological traits of the host insect. There is a high frequency of Wolbachia infection in the leafhopper Yamatotettix flavovittatus Matsumura. To investigate the potential roles of Wolbachia in the host, it is important to generate a non-Wolbachia-infected line. The efficacy of antibiotics in eliminating Wolbachia from Y. flavovittatus remains unknown. This leafhopper harbors the mutualistic bacterium Candidatus Sulcia muelleri, which has an important function in the biological traits. The presence of Ca. S. muelleri raises a major concern regarding the use of antibiotics. We selectively eliminated Wolbachia, considering the influence of antibiotics on leafhopper survival and Ca. S. muelleri prevalence. The effect of artificial diets containing different doses of tetracycline and rifampicin on survival was optimized; high dose (0.5 mg/ml) of antibiotics induces a high mortality. A concentration of 0.2 mg/ml was chosen for the subsequent experiments. Antibiotic treatments significantly reduced the Wolbachia infection, and the Wolbachia density in the treated leafhoppers sharply declined. Wolbachia recurred in tetracycline-treated offspring, regardless of antibiotic exposure. However, Wolbachia is unable to be transmitted and restored in rifampicin-treated offspring. The dose and treatment duration had no significant effect on the infection and density of Ca. S. muelleri in the antibiotic-treated offspring. In conclusion, Wolbachia in Y. flavovittatus was stably eliminated using rifampicin, and the Wolbachia-free line was generated at least two generations after treatment. This report provides additional experimental procedures for removing Wolbachia from insects, particularly in host species with the coexistence of Ca. S. muelleri.}, } @article {pmid35471983, year = {2022}, author = {Martín-Park, A and Che-Mendoza, A and Contreras-Perera, Y and Pérez-Carrillo, S and Puerta-Guardo, H and Villegas-Chim, J and Guillermo-May, G and Medina-Barreiro, A and Delfín-González, H and Méndez-Vales, R and Vázquez-Narvaez, S and Palacio-Vargas, J and Correa-Morales, F and Ayora-Talavera, G and Pavía-Ruz, N and Liang, X and Fu, P and Zhang, D and Wang, X and Toledo-Romaní, ME and Xi, Z and Vázquez-Prokopec, G and Manrique-Saide, P}, title = {Pilot trial using mass field-releases of sterile males produced with the incompatible and sterile insect techniques as part of integrated Aedes aegypti control in Mexico.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {4}, pages = {e0010324}, pmid = {35471983}, issn = {1935-2735}, mesh = {*Aedes ; Animals ; Female ; Humans ; *Infertility, Male ; Insecta ; Male ; Mexico ; Mosquito Control/methods ; Mosquito Vectors ; Pilot Projects ; *Wolbachia ; }, abstract = {BACKGROUND: The combination of Wolbachia-based incompatible insect technique (IIT) and radiation-based sterile insect technique (SIT) can be used for population suppression of Aedes aegypti. Our main objective was to evaluate whether open-field mass-releases of wAlbB-infected Ae. aegypti males, as part of an Integrated Vector Management (IVM) plan led by the Mexican Ministry of Health, could suppress natural populations of Ae. aegypti in urbanized settings in south Mexico.

We implemented a controlled before-and-after quasi-experimental study in two suburban localities of Yucatan (Mexico): San Pedro Chimay (SPC), which received IIT-SIT, and San Antonio Tahdzibichén used as control. Release of wAlbB Ae. aegypti males at SPC extended for 6 months (July-December 2019), covering the period of higher Ae. aegypti abundance. Entomological indicators included egg hatching rates and outdoor/indoor adult females collected at the release and control sites. Approximately 1,270,000 lab-produced wAlbB-infected Ae. aegypti males were released in the 50-ha treatment area (2,000 wAlbB Ae. aegypti males per hectare twice a week in two different release days, totaling 200,000 male mosquitoes per week). The efficacy of IIT-SIT in suppressing indoor female Ae. aegypti density (quantified from a generalized linear mixed model showing a statistically significant reduction in treatment versus control areas) was 90.9% a month after initiation of the suppression phase, 47.7% two months after (when number of released males was reduced in 50% to match local abundance), 61.4% four months after (when initial number of released males was re-established), 88.4% five months after and 89.4% at six months after the initiation of the suppression phase. A proportional, but lower, reduction in outdoor female Ae. aegypti was also quantified (range, 50.0-75.2% suppression).

CONCLUSIONS/SIGNIFICANCE: Our study, the first open-field pilot implementation of Wolbachia IIT-SIT in Mexico and Latin-America, confirms that inundative male releases can significantly reduce natural populations of Ae. aegypti. More importantly, we present successful pilot results of the integration of Wolbachia IIT-SIT within a IVM plan implemented by Ministry of Health personnel.}, } @article {pmid35463816, year = {2022}, author = {Ehrens, A and Hoerauf, A and Hübner, MP}, title = {Current perspective of new anti-Wolbachial and direct-acting macrofilaricidal drugs as treatment strategies for human filariasis.}, journal = {GMS infectious diseases}, volume = {10}, number = {}, pages = {Doc02}, pmid = {35463816}, issn = {2195-8831}, abstract = {Filarial diseases like lymphatic filariasis and onchocerciasis belong to the Neglected Tropical Diseases and remain a public health problem in endemic countries. Lymphatic filariasis and onchocerciasis can lead to stigmatizing pathologies and present a socio-economic burden for affected people and their endemic countries. Current treatment recommendations by the WHO include mass drug administration with ivermectin for the treatment of onchocerciasis and a combination of ivermectin, albendazole and diethylcarbamazine (DEC) for the treatment of lymphatic filariasis in areas that are not co-endemic for onchocerciasis or loiasis. Limitations of these treatment strategies are due to potential severe adverse events in onchocerciasis and loiasis patients following DEC or ivermectin treatment, respectively, the lack of a macrofilaricidal efficacy of those drugs and the risk of drug resistance development. Thus, to achieve the elimination of transmission of onchocerciasis and the elimination of lymphatic filariasis as a public health problem by 2030, the WHO defined in its roadmap that new alternative treatment strategies with macrofilaricidal compounds are required. Within a collaboration of the non-profit organizations Drugs for Neglected Diseases initiative (DNDi), the Bill & Melinda Gates Foundation, and partners from academia and industry, several new promising macrofilaricidal drug candidates were identified, which will be discussed in this review.}, } @article {pmid35458433, year = {2022}, author = {Stica, CJ and Barrero, RA and Murray, RZ and Devine, GJ and Phillips, MJ and Frentiu, FD}, title = {Global Evolutionary History and Dynamics of Dengue Viruses Inferred from Whole Genome Sequences.}, journal = {Viruses}, volume = {14}, number = {4}, pages = {}, pmid = {35458433}, issn = {1999-4915}, mesh = {*Dengue ; *Dengue Virus ; Evolution, Molecular ; Genome, Viral ; Genotype ; Humans ; Phylogeny ; *Wolbachia ; }, abstract = {Dengue is an arboviral disease caused by dengue virus (DENV), leading to approximately 25,000 deaths/year and with over 40% of the world's population at risk. Increased international travel and trade, poorly regulated urban expansion, and warming global temperatures have expanded the geographic range and incidence of the virus in recent decades. This study used phylogenetic and selection pressure analyses to investigate trends in DENV evolution, using whole genome coding sequences from publicly available databases alongside newly sequenced isolates collected between 1963-1997 from Southeast Asia and the Pacific. Results revealed very similar phylogenetic relationships when using the envelope gene and the whole genome coding sequences. Although DENV evolution is predominantly driven by negative selection, a number of amino acid sites undergoing positive selection were found across the genome, with the majority located in the envelope and NS5 genes. Some genotypes appear to be diversifying faster than others within each serotype. The results from this research improve our understanding of DENV evolution, with implications for disease control efforts such as Wolbachia-based biocontrol and vaccine design.}, } @article {pmid35446252, year = {2022}, author = {Quek, S and Cerdeira, L and Jeffries, CL and Tomlinson, S and Walker, T and Hughes, GL and Heinz, E}, title = {Wolbachia endosymbionts in two Anopheles species indicates independent acquisitions and lack of prophage elements.}, journal = {Microbial genomics}, volume = {8}, number = {4}, pages = {}, pmid = {35446252}, issn = {2057-5858}, support = {BB/V011278/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 217303/Z/19/Z/WT_/Wellcome Trust/United Kingdom ; 101285/WT_/Wellcome Trust/United Kingdom ; BB/T001240/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; R01 AI116811/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Anopheles ; Prophages/genetics ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a genus of obligate bacterial endosymbionts that infect a diverse range of arthropod species as well as filarial nematodes, with its single described species, Wolbachia pipientis , divided into several ‘supergroups’ based on multilocus sequence typing. Wolbachia strains in mosquitoes have been shown to inhibit the transmission of human pathogens, including Plasmodium malaria parasites and arboviruses. Despite their large host range, Wolbachia strains within the major malaria vectors of the Anopheles gambiae and Anopheles funestus complexes appear at low density, established solely on PCR-based methods. Questions have been raised as to whether this represents a true endosymbiotic relationship. However, recent definitive evidence for two distinct, high-density strains of supergroup B Wolbachia within Anopheles demeilloni and Anopheles moucheti has opened exciting possibilities to explore naturally occurring Wolbachia endosymbionts in Anopheles for biocontrol strategies to block Plasmodium transmission. Here, we utilize genomic analyses to demonstrate that both Wolbachia strains have retained all key metabolic and transport pathways despite their smaller genome size, with this reduction potentially attributable to degenerated prophage regions. Even with this reduction, we confirmed the presence of cytoplasmic incompatibility (CI) factor genes within both strains, with wAnD maintaining intact copies of these genes while the cifB gene was interrupted in wAnM, so functional analysis is required to determine whether wAnM can induce CI. Additionally, phylogenetic analysis indicates that these Wolbachia strains may have been introduced into these two Anopheles species via horizontal transmission events, rather than by ancestral acquisition and subsequent loss events in the Anopheles gambiae species complex. These are the first Wolbachia genomes, to our knowledge, that enable us to study the relationship between natural strain Plasmodium malaria parasites and their anopheline hosts.}, } @article {pmid35442957, year = {2022}, author = {Tantowijoyo, W and Tanamas, SK and Nurhayati, I and Setyawan, S and Budiwati, N and Fitriana, I and Ernesia, I and Wardana, DS and Supriyati, E and Arguni, E and Meitika, Y and Prabowo, E and Andari, B and Green, BR and Hodgson, L and Rancès, E and Ryan, PA and O'Neill, SL and Anders, KL and Ansari, MR and Indriani, C and Ahmad, RA and Utarini, A and Simmons, CP}, title = {Aedes aegypti abundance and insecticide resistance profiles in the Applying Wolbachia to Eliminate Dengue trial.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {4}, pages = {e0010284}, pmid = {35442957}, issn = {1935-2735}, mesh = {*Aedes ; Animals ; *Dengue/epidemiology/prevention & control ; *Dengue Virus ; Insecticide Resistance ; Mosquito Vectors ; *Wolbachia ; }, abstract = {The Applying Wolbachia to Eliminate Dengue (AWED) trial was a parallel cluster randomised trial that demonstrated Wolbachia (wMel) introgression into Ae. aegypti populations reduced dengue incidence. In this predefined substudy, we compared between treatment arms, the relative abundance of Ae. aegypti and Ae. albopictus before, during and after wMel-introgression. Between March 2015 and March 2020, 60,084 BG trap collections yielded 478,254 Ae. aegypti and 17,623 Ae. albopictus. Between treatment arms there was no measurable difference in Ae. aegypti relative abundance before or after wMel-deployments, with a count ratio of 0.96 (95% CI 0.76, 1.21) and 1.00 (95% CI 0.85, 1.17) respectively. More Ae. aegypti were caught per trap per week in the wMel-intervention arm compared to the control arm during wMel deployments (count ratio 1.23 (95% CI 1.03, 1.46)). Between treatment arms there was no measurable difference in the Ae. albopictus population size before, during or after wMel-deployment (overall count ratio 1.10 (95% CI 0.89, 1.35)). We also compared insecticide resistance phenotypes of Ae. aegypti in the first and second years after wMel-deployments. Ae. aegypti field populations from wMel-treated and untreated arms were similarly resistant to malathion (0.8%), permethrin (1.25%) and cyfluthrin (0.15%) in year 1 and year 2 of the trial. In summary, we found no between-arm differences in the relative abundance of Ae. aegypti or Ae. albopictus prior to or after wMel introgression, and no between-arm difference in Ae. aegypti insecticide resistance phenotypes. These data suggest neither Aedes abundance, nor insecticide resistance, confounded the epidemiological outcomes of the AWED trial.}, } @article {pmid35437949, year = {2022}, author = {Zhou, JC and Shang, D and Qian, Q and Zhang, C and Zhang, LS and Dong, H}, title = {Penetrance during Wolbachia-mediated parthenogenesis of Trichogramma wasps is reduced by continuous oviposition, associated with exhaustion of Wolbachia titers in ovary and offspring eggs.}, journal = {Pest management science}, volume = {78}, number = {7}, pages = {3080-3089}, doi = {10.1002/ps.6934}, pmid = {35437949}, issn = {1526-4998}, support = {//Agricultural Science and Technology Innovation Program (CAAS-ZDRW202108)/ ; //Fundamental Research Funds for the Universities of Liaoning Province (LR2019061, LJKZ0646)/ ; //Major Projects of China National Tobacco Corporation (110202001032 (LS-01))/ ; //Natural Science Foundation of Liaoning Province (2020-BS-137)/ ; //Projects of Guizhou Tobacco Corporation (201936, 201937, and 201941)/ ; //Projects of National Natural Science Foundation of China (32102275, 32172476)/ ; //Natural Science Foundation of Liaoning Province/ ; //Agricultural Science and Technology Innovation Program/ ; //China National Tobacco Corporation/ ; //National Natural Science Foundation of China/ ; }, mesh = {Animals ; Female ; In Situ Hybridization, Fluorescence ; Ovary ; Oviposition ; Parthenogenesis ; Penetrance ; *Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Thelytokous Wolbachia-infected Trichogramma wasps are superior to bisexual uninfected wasps regarding biological control programs. However, continuous oviposition weakens the parthenogenesis-inducing (PI) strength of Wolbachia. Whether this reduced PI strength relates to decreases in the titer of Wolbachia in the ovary and offspring eggs of Trichogramma remains unclear. Here, using fluorescence in situ hybridization (FISH) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) methods, we investigated how the penetrance of Wolbachia-mediated parthenogenesis, Wolbachia density, and distributions of two Wolbachia-infected Trichogramma species, T. pretiosum (TP) and T. dendrolimi (TD), were influenced by different host access treatments [newly-emerged virgin females (NE), 7-day-old females without access to host eggs (NAH), and 7-day-old virgin females with access to host eggs (AH)].

RESULTS: Continuous oviposition decreased Wolbachia PI strength and titers in TP and TD. Continuous oviposition in AH decreased Wolbachia titers in abdomen and offspring eggs of TP and TD females, compared with NAH and NE; NAH had a lower thorax Wolbachia titer than NE. The numbers of parasitized host eggs and offspring wasps, and emergence rates of offspring deposited by AH were lower than those of NE and NAH, for either species.

CONCLUSION: Weakened PI strength, driven by continuous oviposition in Trichogramma wasps, is associated with Wolbachia titer exhaustion in ovary and offspring eggs. Wolbachia density is dependent on PI strength in Trichogramma wasps, highlighting the side effects of continuous oviposition regarding thelytokous Wolbachia-infected Trichogramma in biological control programs. © 2022 Society of Chemical Industry.}, } @article {pmid35432921, year = {2022}, author = {Darwell, CT and Souto-Vilarós, D and Michalek, J and Boutsi, S and Isua, B and Sisol, M and Kuyaiva, T and Weiblen, G and Křivan, V and Novotny, V and Segar, ST}, title = {Predicting distributions of Wolbachia strains through host ecological contact-Who's manipulating whom?.}, journal = {Ecology and evolution}, volume = {12}, number = {4}, pages = {e8826}, pmid = {35432921}, issn = {2045-7758}, abstract = {Reproductive isolation in response to divergent selection is often mediated via third-party interactions. Under these conditions, speciation is inextricably linked to ecological context. We present a novel framework for understanding arthropod speciation as mediated by Wolbachia, a microbial endosymbiont capable of causing host cytoplasmic incompatibility (CI). We predict that sympatric host sister-species harbor paraphyletic Wolbachia strains that provide CI, while well-defined congeners in ecological contact and recently diverged noninteracting congeners are uninfected due to Wolbachia redundancy. We argue that Wolbachia provides an adaptive advantage when coupled with reduced hybrid fitness, facilitating assortative mating between co-occurring divergent phenotypes-the contact contingency hypothesis. To test this, we applied a predictive algorithm to empirical pollinating fig wasp data, achieving up to 91.60% accuracy. We further postulate that observed temporal decay of Wolbachia incidence results from adaptive host purging-adaptive decay hypothesis-but implementation failed to predict systematic patterns. We then account for post-zygotic offspring mortality during CI mating, modeling fitness clines across developmental resources-the fecundity trade-off hypothesis. This model regularly favored CI despite fecundity losses. We demonstrate that a rules-based algorithm accurately predicts Wolbachia infection status. This has implications among other systems where closely related sympatric species encounter adaptive disadvantage through hybridization.}, } @article {pmid35420439, year = {2022}, author = {Rohkin Shalom, S and Weiss, B and Lalzar, M and Kaltenpoth, M and Chiel, E}, title = {Abundance and Localization of Symbiotic Bacterial Communities in the Fly Parasitoid Spalangia cameroni.}, journal = {Applied and environmental microbiology}, volume = {88}, number = {9}, pages = {e0254921}, pmid = {35420439}, issn = {1098-5336}, mesh = {Animals ; Enterobacteriaceae/genetics ; Female ; *Gammaproteobacteria ; In Situ Hybridization, Fluorescence ; Male ; *Rickettsia/genetics ; Symbiosis/physiology ; *Wasps/microbiology ; *Wolbachia/physiology ; }, abstract = {Multicellular eukaryotes often host multiple microbial symbionts that may cooperate or compete for host resources, such as space and nutrients. Here, we studied the abundances and localization of four bacterial symbionts, Rickettsia, Wolbachia, Sodalis, and Arsenophonus, in the parasitic wasp Spalangia cameroni. Using quantitative PCR (qPCR), we measured the symbionts' titers in wasps that harbor different combinations of these symbionts. We found that the titer of each symbiont decreased as the number of symbiont species in the community increased. Symbionts' titers were higher in females than in males. Rickettsia was the most abundant symbiont in all the communities, followed by Sodalis and Wolbachia. The titers of these three symbionts were positively correlated in some of the colonies. Fluorescence in situ hybridization was in line with the qPCR results: Rickettsia, Wolbachia, and Sodalis were observed in high densities in multiple organs, including brain, muscles, gut, Malpighian tubules, fat body, ovaries, and testes, while Arsenophonus was localized to fewer organs and in lower densities. Sodalis and Arsenophonus were observed in ovarian follicle cells but not within oocytes or laid eggs. This study highlights the connection between symbionts' abundance and localization. We discuss the possible connections between our findings to symbiont transmission success. IMPORTANCE Many insects carry intracellular bacterial symbionts (bacteria that reside within the cells of the insect). When multiple symbiont species cohabit in a host, they may compete or cooperate for space, nutrients, and transmission, and the nature of such interactions would be reflected in the abundance of each symbiont species. Given the widespread occurrence of coinfections with maternally transmitted symbionts in insects, it is important to learn more about how they interact, where they are localized, and how these two aspects affect their co-occurrence within individual insects. Here, we studied the abundance and the localization of four symbionts, Rickettsia, Wolbachia, Sodalis, and Arsenophonus, that cohabit the parasitic wasp Spalangia cameroni. We found that symbionts' titers differed between symbiotic communities. These results were corroborated by microscopy, which shows differential localization patterns. We discuss the findings in the contexts of community ecology, possible symbiont-symbiont interactions, and host control mechanisms that may shape the symbiotic community structure.}, } @article {pmid35417447, year = {2022}, author = {Yang, Q and Chung, J and Robinson, KL and Schmidt, TL and Ross, PA and Liang, J and Hoffmann, AA}, title = {Sex-specific distribution and classification of Wolbachia infections and mitochondrial DNA haplogroups in Aedes albopictus from the Indo-Pacific.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {4}, pages = {e0010139}, pmid = {35417447}, issn = {1935-2735}, mesh = {*Aedes/genetics ; Animals ; DNA, Mitochondrial/genetics ; *Dengue/epidemiology ; Female ; Male ; Mosquito Vectors/genetics ; *Wolbachia/genetics ; }, abstract = {The arbovirus vector Aedes albopictus (Asian tiger mosquito) is common throughout the Indo-Pacific region, where most global dengue transmission occurs. We analysed population genomic data and tested for cryptic species in 160 Ae. albopictus sampled from 16 locations across this region. We found no evidence of cryptic Ae. albopictus but found multiple intraspecific COI haplotypes partitioned into groups representing three Asian lineages: East Asia, Southeast Asia and Indonesia. Papua New Guinea (PNG), Vanuatu and Christmas Island shared recent coancestry, and Indonesia and Timor-Leste were likely invaded from East Asia. We used a machine learning trained on morphologically sexed samples to classify sexes using multiple genetic features and then characterized the wAlbA and wAlbB Wolbachia infections in 664 other samples. The wAlbA and wAlbB infections as detected by qPCR showed markedly different patterns in the sexes. For females, most populations had a very high double infection incidence, with 67% being the lowest value (from Timor-Leste). For males, the incidence of double infections ranged from 100% (PNG) to 0% (Vanuatu). Only 6 females were infected solely by the wAlbA infection, while rare uninfected mosquitoes were found in both sexes. The wAlbA and wAlbB densities varied significantly among populations. For mosquitoes from Torres Strait and Vietnam, the wAlbB density was similar in single-infected and superinfected (wAlbA and wAlbB) mosquitoes. There was a positive association between wAlbA and wAlbB infection densities in superinfected Ae. albopictus. Our findings provide no evidence of cryptic species of Ae. albopictus in the region and suggest site-specific factors influencing the incidence of Wolbachia infections and their densities. We also demonstrate the usefulness of ddRAD tag depths as sex-specific mosquito markers. The results provide baseline data for the exploitation of Wolbachia-induced cytoplasmic incompatibility (CI) in dengue control.}, } @article {pmid35417002, year = {2022}, author = {Tamarozzi, F and Rodari, P and Salas-Coronas, J and Bottieau, E and Salvador, F and Soriano-Pérez, MJ and Cabeza-Barrera, MI and Van Esbroeck, M and Treviño, B and Buonfrate, D and Gobbi, FG}, title = {A large case series of travel-related Mansonella perstans (vector-borne filarial nematode): a TropNet study in Europe.}, journal = {Journal of travel medicine}, volume = {29}, number = {7}, pages = {}, pmid = {35417002}, issn = {1708-8305}, support = {//Ministry of Health/ ; }, mesh = {Animals ; Humans ; Mansonella ; *Mansonelliasis/diagnosis/drug therapy/epidemiology ; Retrospective Studies ; Travel ; Mebendazole/therapeutic use ; Prospective Studies ; Travel-Related Illness ; *Wolbachia ; }, abstract = {BACKGROUND: Infection with Mansonella perstans is a neglected filariasis, widely distributed in sub-Saharan Africa, characterized by an elusive clinical picture; treatment for mansonellosis is not standardized. This retrospective study aimed to describe the clinical features, treatment schemes and evolution, of a large cohort of imported cases of M. perstans infection seen in four European centres for tropical diseases.

METHODS: Mansonella perstans infections, diagnosed by identification of blood microfilariae in migrants, expatriates and travellers, collected between 1994 and 2018, were retrospectively analysed. Data concerning demographics, clinical history and laboratory examinations at diagnosis and at follow-up time points were retrieved.

RESULTS: A total of 392 patients were included in the study. Of the 281 patients for whom information on symptoms could be retrieved, 150 (53.4%) reported symptoms, abdominal pain and itching being the most frequent. Positive serology and eosinophilia were present in 84.4% and 66.1%, respectively, of those patients for whom these data were available. Concomitant parasitic infections were reported in 23.5% of patients. Treatment, administered to 325 patients (82.9%), was extremely heterogeneous between and within centres; the most commonly used regimen was mebendazole 100 mg twice a day for 1 month. A total of 256 (65.3%) patients attended a first follow-up, median 3 months (interquartile range 2-12) after the first visit; 83.1% of patients having received treatment based on mebendazole and/or doxycycline, targeting Wolbachia, became amicrofilaremic, 41.1-78.4% of whom within 12 months from single treatment.

CONCLUSIONS: Lack of specific symptoms, together with the inconstant positivity of parasitological and antibody-based assays in the infected population, makes the clinical suspicion and screening for mansonellosis particularly difficult. Prospective studies evaluating prevalence of infection in migrants from endemic areas, infection-specific morbidity, presence of Wolbachia endosymbionts in M. perstans populations from different geographical areas and efficacy of treatment regimens are absolutely needed to optimize the clinical management of infection.}, } @article {pmid35414231, year = {2022}, author = {Hornett, EA and Kageyama, D and Hurst, GDD}, title = {Sex determination systems as the interface between male-killing bacteria and their hosts.}, journal = {Proceedings. Biological sciences}, volume = {289}, number = {1972}, pages = {20212781}, pmid = {35414231}, issn = {1471-2954}, mesh = {Animals ; *Arthropods/microbiology ; Bacteria/genetics ; Male ; Sex Ratio ; Symbiosis ; *Wolbachia/physiology ; }, abstract = {Arthropods host a range of sex-ratio-distorting selfish elements, including diverse maternally inherited endosymbionts that solely kill infected males. Male-killing heritable microbes are common, reach high frequency, but until recently have been poorly understood in terms of the host-microbe interaction. Additionally, while male killing should generate strong selection for host resistance, evidence of this has been scant. The interface of the microbe with host sex determination is integral to the understanding of how death is sex limited and how hosts can evolve evasion of male killing. We first review current knowledge of the mechanisms diverse endosymbionts use to induce male-specific death. We then examine recent evidence that these agents do produce intense selection for host nuclear suppressor elements. We argue, from our understanding of male-killing mechanisms, that suppression will commonly involve evolution of the host sex determination pathways and that the host's response to male-killing microbes thus represents an unrecognized driver of the diversity of arthropod sex determination. Further work is required to identify the genes and mechanisms responsible for male-killing suppression, which will both determine the components of sex determination (or other) systems associated with suppressor evolution, and allow insight into the mechanism of male killing itself.}, } @article {pmid35413938, year = {2022}, author = {Mejia, AJ and Dutra, HLC and Jones, MJ and Perera, R and McGraw, EA}, title = {Cross-tissue and generation predictability of relative Wolbachia densities in the mosquito Aedes aegypti.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {128}, pmid = {35413938}, issn = {1756-3305}, support = {R01 AI151166/AI/NIAID NIH HHS/United States ; R01 151166//National Institute of Allergy and Infectious Diseases/ ; }, mesh = {*Aedes/physiology ; Animals ; Female ; Humans ; Male ; Mosquito Vectors/physiology ; Specific Gravity ; *Wolbachia/physiology ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {BACKGROUND: The insect endosymbiotic bacterium Wolbachia is being deployed in field populations of the mosquito Aedes aegypti for biological control. This microbe prevents the replication of human disease-causing viruses inside the vector, including dengue, Zika and chikungunya. Relative Wolbachia densities may in part predict the strength of this 'viral blocking' effect. Additionally, Wolbachia densities may affect the strength of the reproductive manipulations it induces, including cytoplasmic incompatibility (CI), maternal inheritance rates or induced fitness effects in the insect host. High rates of CI and maternal inheritance and low rates of fitness effects are also key to the successful spreading of Wolbachia through vector populations and its successful use in biocontrol. The factors that control Wolbachia densities are not completely understood.

METHODS: We used quantitative PCR-based methods to estimate relative density of the Wolbachia wAlbB strain in both the somatic and reproductive tissues of adult male and female mosquitoes, as well as in eggs. Using correlation analyses, we assessed whether densities in one tissue predict those in others within the same individual, but also across generations.

RESULTS: We found little relationship among the relative Wolbachia densities of different tissues in the same host. The results also show that there was very little relationship between Wolbachia densities in parents and those in offspring, both in the same and different tissues. The one exception was with ovary-egg relationships, where there was a strong positive association. Relative Wolbachia densities in reproductive tissues were always greater than those in the somatic tissues. Additionally, the densities were consistent in females over their lifetime regardless of tissue, whereas they were generally higher and more variable in males, particularly in the testes.

CONCLUSIONS: Our results indicate that either stochastic processes or local tissue-based physiologies are more likely factors dictating Wolbachia densities in Ae. aegypti individuals, rather than shared embryonic environments or heritable genetic effects of the mosquito genome. These findings have implications for understanding how relative Wolbachia densities may evolve and/or be maintained over the long term in Ae. aegypti.}, } @article {pmid35400954, year = {2022}, author = {Thengchaisri, N and Inpankaew, T and Arthitwong, S and Steiner, JM and Sattasathuchana, P}, title = {Molecular prevalence of Dirofilaria immitis and Wolbachia infections in pet and semi-domesticated cats in Bangkok, Thailand.}, journal = {Veterinary world}, volume = {15}, number = {2}, pages = {239-243}, pmid = {35400954}, issn = {0972-8988}, abstract = {BACKGROUND AND AIM: Although cats are not natural hosts for heartworm infections (Dirofilaria immitis), evidence suggests that feline heartworm disease can be detrimental because of a severe inflammatory response. Recent studies have found that infection with bacteria of the genus Wolbachia is the principal cause of acute inflammatory filaria disease; nonetheless, the prevalence of cats naturally infected with heartworms and Wolbachia remains unclear. This study aimed to estimate the prevalence and current distribution of feline heartworm disease and its association with Wolbachia infection in pet and semi-domesticated cats in Bangkok, Thailand.

MATERIALS AND METHODS: A total of 260 cats (130 pet cats and 130 semi-domesticated cats) were enrolled in this study. Blood samples were placed into ethylenediaminetetraacetic acid tubes for hematological analysis and DNA extraction. A polymerase chain reaction (PCR) was performed to analyze samples for the presence of D. immitis and Wolbachia infections.

RESULTS: The prevalence (95% confidence interval [CI]) of D. immitis infection in pet, semi-domesticated, and all cats were 3.9% (1.3-8.8%), 27.7% (20.2-36.2%), and 19.6% (15.0-25.0%), respectively. The prevalence (95% CI) of Wolbachia infection in pet, semi-domesticated, and all cats were 18.5% (12.2-26.2%), 31.5% (23.7-40.3%), and 25.0% (19.9-30.7%), respectively. The prevalence of D. immitis and Wolbachia infections in semi-domesticated cats was significantly higher than in pet cats (p=0.002 and p=0.022, respectively). There was a significant association between D. immitis and Wolbachia infections (p<0.001). There was also a significant association between D. immitis infection and the presence of eosinophilia (p<0.045).

CONCLUSION: From the PCR analysis, it can be concluded that semi-domesticated cats were at higher risk for D. immitis infection than pet cats. There was a significant association between positive D. immitis infection and positive Wolbachia infection. Combinations of anthelmintic and antimicrobial therapy should be considered in heartworm-positive cats.}, } @article {pmid35381817, year = {2021}, author = {Mohanty, I and Rath, A and Pradhan, N and Panda, BB and Mohapatra, PK and Hazra, RK}, title = {Prevalence and transmission potential of Wolbachia in Aedes albopictus population circulating in endemic coastal districts of Odisha, India.}, journal = {Journal of vector borne diseases}, volume = {58}, number = {4}, pages = {297-305}, doi = {10.4103/0972-9062.313967}, pmid = {35381817}, issn = {0972-9062}, mesh = {*Aedes ; Animals ; *Dengue/epidemiology/prevention & control ; Mosquito Vectors ; Prevalence ; *Wolbachia/genetics ; }, abstract = {Wolbachia, known for its reproductive manipulation capabilities in insects, are being implemented to control dengue and chikungunya. To understand Wolbachia biology and its utility as a bio-control for vector mosquito's populations, we investigated its dissemination pattern in field in collected Ae. albopictus along with its maternal transmission efficacy over generations in regions of endemic dengue (DENV) transmission. Field collected Ae. albopictus were subjected to PCR for Wolbachia screening. Overall mean Wolbachia infection frequency in Ae. albopictus was found out to be 87.3% wherein a trend was observed in the pattern of maternal transmission across generations. χ[2] for trend revealed a significant variation between Wolbachia infections and non-infections in Ae. albopictus generations. Linear regression analysis revealed the involvement of a strong negative correlation, implying that overall Wolbachia infection tends to decrease in places with high dengue cases.The reduction in Wolbachia infection frequency may be attributed to several environmental factors with the probability of being the cause for endemicity of dengue in the studied areas.This study reports on the transmission efficacy of naturally occurring Wolbachia in successive generations of Ae. albopictus and its correlation with dengue cases in clusters of Odisha, India. Studying the transmission trend of Wolbachia along with transovarial transmission of DENV might be indicative towards the interplay of Wolbachia infection in presence/absence of DENV.}, } @article {pmid35377795, year = {2022}, author = {Quek, S and Cook, DAN and Wu, Y and Marriott, AE and Steven, A and Johnston, KL and Ford, L and Archer, J and Hemingway, J and Ward, SA and Wagstaff, SC and Turner, JD and Taylor, MJ}, title = {Wolbachia depletion blocks transmission of lymphatic filariasis by preventing chitinase-dependent parasite exsheathment.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {119}, number = {15}, pages = {e2120003119}, pmid = {35377795}, issn = {1091-6490}, support = {MC_PC_18055/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; *Anti-Bacterial Agents/pharmacology ; *Chitinases/genetics ; *Elephantiasis, Filarial/transmission ; Humans ; *Microfilariae/enzymology/growth & development/microbiology ; Mosquito Vectors/parasitology ; *Wolbachia/drug effects/genetics ; }, abstract = {Lymphatic filariasis is a vector-borne neglected tropical disease prioritized for global elimination. The filarial nematodes that cause the disease host a symbiotic bacterium, Wolbachia, which has been targeted using antibiotics, leading to cessation of parasite embryogenesis, waning of circulating larvae (microfilariae [mf]), and gradual cure of adult infection. One of the benefits of the anti-Wolbachia mode of action is that it avoids the rapid killing of mf, which can drive inflammatory adverse events. However, mf depleted of Wolbachia persist for several months in circulation, and thus patients treated with antibiotics are assumed to remain at risk for transmitting infections. Here, we show that Wolbachia-depleted mf rapidly lose the capacity to develop in the mosquito vector through a defect in exsheathment and inability to migrate through the gut wall. Transcriptomic and Western blotting analyses demonstrate that chitinase, an enzyme essential for mf exsheathment, is down-regulated in Wolbachia-depleted mf and correlates with their inability to exsheath and escape the mosquito midgut. Supplementation of in vitro cultures of Wolbachia-depleted mf with chitinase enzymes restores their ability to exsheath to a similar level to that observed in untreated mf. Our findings elucidate a mechanism of rapid transmission-blocking activity of filariasis after depletion of Wolbachia and adds to the broad range of biological processes of filarial nematodes that are dependent on Wolbachia symbiosis.}, } @article {pmid35373850, year = {2022}, author = {Ajendra, J and Allen, JE}, title = {Neutrophils: Friend or foe in Filariasis?.}, journal = {Parasite immunology}, volume = {44}, number = {6}, pages = {e12918}, doi = {10.1111/pim.12918}, pmid = {35373850}, issn = {1365-3024}, support = {MR/V011235/1/MRC_/Medical Research Council/United Kingdom ; 106898/A/15/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Elephantiasis, Filarial ; *Filarioidea ; Humans ; Immunity ; Mice ; Neutrophils ; *Wolbachia ; }, abstract = {Infection with the filarial nematodes that cause diseases such as lymphatic filariasis and onchocerciasis represent major public health challenges. With millions of people at risk of infection, new strategies for treatment or prevention are urgently needed. More complete understanding of the host immune system's ability to control and eliminate the infection is an important step towards fighting these debilitating infectious diseases. Neutrophils are innate immune cells that are rapidly recruited to inflamed or infected tissues and while considered primarily anti-microbial, there is increasing recognition of their role in helminth infections. Filarial nematodes present a unique situation, as many species harbour the bacterial endosymbiont, Wolbachia. The unexpected involvement of neutrophils during filarial infections has been revealed both in human diseases and animal studies, with strong evidence for recruitment by Wolbachia. This present review will introduce the different human filarial diseases and discuss neutrophil involvement in both protective immune responses, but also in the exacerbation of pathology. Additionally, we will highlight the contributions of the murine model of filariasis, Litomosoides sigmodontis. While several studies have revealed the importance of neutrophils in these parasite infections, we will also draw attention to many questions that remain to be answered.}, } @article {pmid35369485, year = {2022}, author = {Hussain, S and Perveen, N and Hussain, A and Song, B and Aziz, MU and Zeb, J and Li, J and George, D and Cabezas-Cruz, A and Sparagano, O}, title = {The Symbiotic Continuum Within Ticks: Opportunities for Disease Control.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {854803}, pmid = {35369485}, issn = {1664-302X}, abstract = {Among blood-sucking arthropods, ticks are recognized as being of prime global importance because of their role as vectors of pathogens affecting human and animal health. Ticks carry a variety of pathogenic, commensal, and symbiotic microorganisms. For the latter, studies are available concerning the detection of endosymbionts, but their role in the physiology and ecology of ticks remains largely unexplored. This review paper focuses on tick endosymbionts of the genera Coxiella, Rickettsia, Francisella, Midichloria, and Wolbachia, and their impact on ticks and tick-pathogen interactions that drive disease risk. Tick endosymbionts can affect tick physiology by influencing nutritional adaptation, fitness, and immunity. Further, symbionts may influence disease ecology, as they interact with tick-borne pathogens and can facilitate or compete with pathogen development within the vector tissues. Rickettsial symbionts are frequently found in ticks of the genera of Ixodes, Amblyomma, and Dermacentor with relatively lower occurrence in Rhipicephalus, Haemaphysalis, and Hyalomma ticks, while Coxiella-like endosymbionts (CLEs) were reported infecting almost all tick species tested. Francisella-like endosymbionts (FLEs) have been identified in tick genera such as Dermacentor, Amblyomma, Ornithodoros, Ixodes, and Hyalomma, whereas Wolbachia sp. has been detected in Ixodes, Amblyomma, Hyalomma, and Rhipicephalus tick genera. Notably, CLEs and FLEs are obligate endosymbionts essential for tick survival and development through the life cycle. American dog ticks showed greater motility when infected with Rickettsia, indirectly influencing infection risk, providing evidence of a relationship between tick endosymbionts and tick-vectored pathogens. The widespread occurrence of endosymbionts across the tick phylogeny and evidence of their functional roles in ticks and interference with tick-borne pathogens suggests a significant contribution to tick evolution and/or vector competence. We currently understand relatively little on how these endosymbionts influence tick parasitism, vector capacity, pathogen transmission and colonization, and ultimately on how they influence tick-borne disease dynamics. Filling this knowledge gap represents a major challenge for future research.}, } @article {pmid35364056, year = {2022}, author = {Andreychuk, S and Yakob, L}, title = {Mathematical modelling to assess the feasibility of Wolbachia in malaria vector biocontrol.}, journal = {Journal of theoretical biology}, volume = {542}, number = {}, pages = {111110}, doi = {10.1016/j.jtbi.2022.111110}, pmid = {35364056}, issn = {1095-8541}, mesh = {*Aedes ; Animals ; *Anopheles ; Feasibility Studies ; *Malaria ; Models, Theoretical ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Releasing mosquitoes transinfected with the endosymbiotic bacterium Wolbachia is a novel strategy for interrupting vector-borne pathogen transmission. Following its success in controlling arboviruses spread by Aedes aegypti, this technology is being adapted for anopheline malaria vectors. However, antagonistic interactions between Wolbachia and naturally resident Asaia bacteria in malaria vectors have been demonstrated experimentally, potentially jeopardising Wolbachia biocontrol. We developed the first mathematical model accounting for interspecific competition between endosymbionts to assess the feasibility of this novel strategy for controlling malaria. First, Asaia prevalences among natural mosquito populations were compared with simulations parametrized with rates of Asaia transmission reported from laboratory studies. Discrepancies between projections and natural Asaia prevalences indicated potential overestimation of Asaia transmissibility in artificial laboratory settings. With parametrization that matches natural Asaia prevalence, simulations identified redundancies in Asaia's many infection routes (vertical, sexual and environmental). This resilience was only overcome when Wolbachia conferred very high resistance to environmental infection with Asaia, resulting in Wolbachia fixation and Asaia exclusion. Wolbachia's simulated spread was prevented when its maternal transmission was impeded in coinfected mosquitoes and the pre-control Asaia prevalence was beyond a threshold of 60-75%. This theoretical assessment highlights critical next steps in laboratory experiments to inform this strategy's feasibility.}, } @article {pmid35359740, year = {2022}, author = {Salgueiro, J and Nussenbaum, AL and Milla, FH and Asimakis, E and Goane, L and Ruiz, MJ and Bachmann, GE and Vera, MT and Stathopoulou, P and Bourtzis, K and Deutscher, AT and Lanzavecchia, SB and Tsiamis, G and Segura, DF}, title = {Analysis of the Gut Bacterial Community of Wild Larvae of Anastrepha fraterculus sp. 1: Effect of Host Fruit, Environment, and Prominent Stable Associations of the Genera Wolbachia, Tatumella, and Enterobacter.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {822990}, pmid = {35359740}, issn = {1664-302X}, abstract = {The genus Anastrepha (Diptera Tephritidae) includes some of the most important fruit fly pests in the Americas. Here, we studied the gut bacterial community of 3rd instar larvae of Anastrepha fraterculus sp. 1 through Next Generation Sequencing (lllumina) of the V3-V4 hypervariable region within the 16S rRNA gene. Gut bacterial communities were compared between host species (guava and peach), and geographical origins (Concordia and Horco Molle in Argentina) representing distinct ecological scenarios. In addition, we explored the effect of spatial scale by comparing the samples collected from different trees within each geographic origin and host species. We also addressed the effect of fruit size on bacterial diversity. The gut bacterial community was affected both by host species and geographic origin. At smaller spatial scales, the gut bacterial profile differed among trees of the same species and location at least in one host-location combination. There was no effect of fruit size on the larval gut bacteriome. Operational Taxonomic Units (OTUs) assigned to Wolbachia, Tatumella and Enterobacter were identified in all samples examined, which suggest potential, non-transient symbioses. Better knowledge on the larval gut bacteriome contributes valuable information to develop sustainable control strategies against A. fraterculus targeting key symbionts as the Achilles' heel to control this important fruit fly pest.}, } @article {pmid35357208, year = {2022}, author = {Strunov, A and Schmidt, K and Kapun, M and Miller, WJ}, title = {Restriction of Wolbachia Bacteria in Early Embryogenesis of Neotropical Drosophila Species via Endoplasmic Reticulum-Mediated Autophagy.}, journal = {mBio}, volume = {13}, number = {2}, pages = {e0386321}, pmid = {35357208}, issn = {2150-7511}, support = {P 32275/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Autophagy ; Drosophila/microbiology ; Embryonic Development ; Endoplasmic Reticulum ; *Wolbachia/genetics ; }, abstract = {Wolbachia are maternally transmitted intracellular bacteria that are not only restricted to the reproductive organs but also found in various somatic tissues of their native hosts. The abundance of the endosymbiont in the soma, usually a dead end for vertically transmitted bacteria, causes a multitude of effects on life history traits of their hosts, which are still not well understood. Thus, deciphering the host-symbiont interactions on a cellular level throughout a host's life cycle is of great importance to understand their homeostatic nature, persistence, and spreading success. Using fluorescent and transmission electron microscopy, we conducted a comprehensive analysis of Wolbachia tropism in soma and germ line of six Drosophila species at the intracellular level during host development. Our data uncovered diagnostic patterns of infections to embryonic primordial germ cells and to particular cells of the soma in three different neotropical Drosophila species that have apparently evolved independently. We further found that restricted patterns of Wolbachia tropism are determined in early embryogenesis via selective autophagy, and their spatially restricted infection patterns are preserved in adult flies. We observed tight interactions of Wolbachia with membranes of the endoplasmic reticulum, which might play a scaffolding role for autophagosome formation and subsequent elimination of the endosymbiont. Finally, by analyzing D. simulans lines transinfected with nonnative Wolbachia, we uncovered that the host genetic background regulates tissue tropism of infection. Our data demonstrate a novel and peculiar mechanism to limit and spatially restrict bacterial infection in the soma during a very early stage of host development. IMPORTANCE All organisms are living in close and intimate interactions with microbes that cause conflicts but also cooperation between both unequal genetic partners due to their different innate interests of primarily enhancing their own fitness. However, stable symbioses often result in homeostatic interaction, named mutualism, by balancing costs and benefits, where both partners profit. Mechanisms that have evolved to balance and stably maintain homeostasis in mutualistic relationships are still quite understudied; one strategy is to "domesticate" potentially beneficial symbionts by actively controlling their replication rate below a critical and, hence, costly threshold, and/or to spatially and temporally restrict their localization in the host organism, which, in the latter case, in its most extreme form, is the formation of a specialized housing organ for the microbe (bacteriome). However, questions remain: how do these mutualistic associations become established in their first place, and what are the mechanisms for symbiont control and restriction in their early stages? Here, we have uncovered an unprecedented symbiont control mechanism in neotropical Drosophila species during early embryogenesis. The fruit fly evolved selective autophagy to restrict and control the proliferation of its intracellular endosymbiont Wolbachia in a defined subset of the stem cells as soon as the host's zygotic genome is activated.}, } @article {pmid35353007, year = {2022}, author = {Li, TP and Zhou, CY and Wang, MK and Zha, SS and Chen, J and Bing, XL and Hoffmann, AA and Hong, XY}, title = {Endosymbionts Reduce Microbiome Diversity and Modify Host Metabolism and Fecundity in the Planthopper Sogatella furcifera.}, journal = {mSystems}, volume = {7}, number = {2}, pages = {e0151621}, pmid = {35353007}, issn = {2379-5077}, mesh = {Animals ; Fertility ; Reproduction ; Bacteroidetes ; *Microbiota ; Nymph ; *Wolbachia ; *Hemiptera ; }, abstract = {Endosymbionts can strongly affect bacterial microbiota in pests. The white-backed planthopper Sogatella furcifera, a notorious pest in rice, is usually co-infected with Cardinium and Wolbachia, but the effects of these endosymbionts together or individually on the host microbiome and fecundity are unclear. Here, we established three S. furcifera lines (Cardinium and Wolbachia double-infected, Cardinium single-infected, and both-uninfected lines) backcrossed to a common nuclear background and found that single and double infections reduced bacterial diversity and changed bacterial community structure across nymph and adult stages and across adult tissues. The endosymbionts differed in densities between adults and nymphs as well as across adult tissues, with the distribution of Cardinium affected by Wolbachia. Both the single infection and particularly the double infection reduced host fecundity. Lines also differed in levels of metabolites, some of which may influence fecundity (e.g., arginine biosynthesis and nicotinamide metabolism). Cardinium in the single-infected line upregulated metabolic levels, while Wolbachia in the double-infected line appeared to mainly downregulate them. Association analysis pointed to possible connections between various bacteria and differential metabolites. These results reveal that Cardinium by itself and in combination with Wolbachia affect bacterial microbiota and levels of metabolites, with likely effects on host fecundity. Many of the effects of these metabolically limited endosymbionts that are dependent on the hosts may be exerted through manipulation of the microbiome. IMPORTANCE Endosymbionts can profoundly affect the nutrition, immunity, development, and reproduction of insect hosts, but the effects of multiple endosymbiont infections on microbiota and the interaction of these effects with insect host fitness are not well known. By establishing S. furcifera lines with different endosymbiont infection status, we found that Cardinium and the combined Cardinium + Wolbachia infections differentially reduced bacterial diversity as well as changing bacterial community structure and affecting metabolism, which may connect to negative fitness effects of the endosymbionts on their host. These results established the connections between reduced bacterial diversity, decreased fecundity and metabolic responses in S. furcifera.}, } @article {pmid35350856, year = {2022}, author = {Katlav, A and Cook, JM and Riegler, M}, title = {Common endosymbionts affect host fitness and sex allocation via egg size provisioning.}, journal = {Proceedings. Biological sciences}, volume = {289}, number = {1971}, pages = {20212582}, pmid = {35350856}, issn = {1471-2954}, mesh = {Animals ; *Arthropods ; Bacteroidetes ; Female ; Humans ; Male ; Reproduction ; Symbiosis ; *Wolbachia ; }, abstract = {It is hard to overemphasize the importance of endosymbionts in arthropod biology, ecology and evolution. Some endosymbionts can complement host metabolic function or provide defence against pathogens; others, such as ubiquitous Wolbachia and Cardinium, have evolved strategies to manipulate host reproduction. A common reproductive manipulation strategy is cytoplasmic incompatibility (CI) between differently infected individuals which can result in female mortality or male development of fertilized eggs in haplodiploid hosts. Recently, an additional role of endosymbionts has been recognized in the modification of sex allocation in sexually reproducing haplodiploids. This was theoretically expected due to the maternal inheritance of endosymbionts and natural selection for them to increase infected female production, yet the underlying mechanism remained unknown. Here, we tested whether and how Cardinium and Wolbachia causing different CI types interact to increase female production in a haplodiploid thrips species where sex allocation depends on both maternal condition and egg size provisioning. We found that Cardinium augmented female production by increasing maternal fitness and egg size, thereby boosting fertilization rate and offspring fitness. Wolbachia, in contrast, reduced the beneficial effects of Cardinium. Our results demonstrate different invasion strategies and antagonistic effects of endosymbiotic bacteria on host fitness and evolution of sex allocation.}, } @article {pmid35349818, year = {2022}, author = {Hochstrasser, M}, title = {Cytoplasmic incompatibility: A Wolbachia toxin-antidote mechanism comes into view.}, journal = {Current biology : CB}, volume = {32}, number = {6}, pages = {R287-R289}, doi = {10.1016/j.cub.2022.02.014}, pmid = {35349818}, issn = {1879-0445}, support = {R35 GM136325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Antidotes ; Cytoplasm ; Cytosol ; Drosophila melanogaster/genetics ; *Wolbachia/genetics ; }, abstract = {The Wolbachia cidA and cidB genes promote bacterial endosymbiont inheritance through the host female germline. CidB is now shown to load into maturing sperm nuclei. Following fertilization, it disrupts paternal chromosome condensation, triggering embryonic arrest if not countered by CidA in Wolbachia-infected eggs.}, } @article {pmid35346758, year = {2022}, author = {Bazzocchi, C and Genchi, M and Lucchetti, C and Cafiso, A and Ciuca, L and McCall, J and Kramer, LH and Vismarra, A}, title = {Transporter gene expression and Wolbachia quantification in adults of Dirofilaria immitis treated in vitro with ivermectin or moxidectin alone or in combination with doxycycline for 12 h.}, journal = {Molecular and biochemical parasitology}, volume = {249}, number = {}, pages = {111475}, doi = {10.1016/j.molbiopara.2022.111475}, pmid = {35346758}, issn = {1872-9428}, mesh = {Animals ; *Dirofilaria immitis/genetics ; *Dog Diseases/drug therapy/parasitology/prevention & control ; Dogs ; Doxycycline/pharmacology/therapeutic use ; Female ; Gene Expression ; Ivermectin/pharmacology ; Macrolides ; Male ; Membrane Transport Proteins/genetics ; *Wolbachia/genetics ; }, abstract = {Due to their marked larvicidal activity, macrocyclic lactones (MLs) are used for the prevention of heartworm disease (Dirofilaria immitis) in dogs. They have also been shown to eliminate adult parasites after long-term administration, with a so-called "slow-kill" effect. In addition, recent studies have established that a combination of doxycycline, which eliminates the endosymbiont Wolbachia, and MLs has superior adulticide effects when compared to MLs alone. It has been hypothesized that the apparent synergism between doxycycline/MLs may be due to interaction with drug efflux transport proteins. The aim of the present study was to evaluate gene expression of several transport proteins in D. immitis adults treated in vitro either with doxycycline alone, ivermectin alone, moxidectin alone, or a combination of ivermectin or moxidectin with doxycycline for 12 h. Quantitative PCR analysis showed a sex-dependent response to treatments. In female worms, Dim-pgp-10, Dim-haf-1 and Dim-haf-5 were upregulated compared to controls with doxycycline alone and when combined with ivermectin. Moxidectin did not induce any changes in gene expression. In males, moxidectin administered alone induced a slight increase in Dim-pgp-10, Dim-pgp-11and Di-avr-14, while ivermectin in combination with doxycycline produced significant upregulation of the ML receptor Di-avr-14. These results suggest possible synergism between the two drug classes and different susceptibility of males vs. females to adulticide effects.}, } @article {pmid35346038, year = {2022}, author = {Queffelec, J and Postma, A and Allison, JD and Slippers, B}, title = {Remnants of horizontal transfers of Wolbachia genes in a Wolbachia-free woodwasp.}, journal = {BMC ecology and evolution}, volume = {22}, number = {1}, pages = {36}, pmid = {35346038}, issn = {2730-7182}, mesh = {Animals ; *Nematoda ; *Pinus ; *Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Wolbachia is a bacterial endosymbiont of many arthropod and nematode species. Due to its capacity to alter host biology, Wolbachia plays an important role in arthropod and nematode ecology and evolution. Sirex noctilio is a woodwasp causing economic loss in pine plantations of the Southern Hemisphere. An investigation into the genome of this wasp revealed the presence of Wolbachia sequences. Due to the potential impact of Wolbachia on the populations of this wasp, as well as its potential use as a biological control agent against invasive insects, this discovery warranted investigation.

RESULTS: In this study we first investigated the presence of Wolbachia in S. noctilio and demonstrated that South African populations of the wasp are unlikely to be infected. We then screened the full genome of S. noctilio and found 12 Wolbachia pseudogenes. Most of these genes constitute building blocks of various transposable elements originating from the Wolbachia genome. Finally, we demonstrate that these genes are distributed in all South African populations of the wasp.

CONCLUSIONS: Our results provide evidence that S. noctilio might be compatible with a Wolbachia infection and that the bacteria could potentially be used in the future to regulate invasive populations of the wasp. Understanding the mechanisms that led to a loss of Wolbachia infection in S. noctilio could indicate which host species or host population should be sampled to find a Wolbachia strain that could be used as a biological control against S. noctilio.}, } @article {pmid35338196, year = {2022}, author = {Wendt, M and Kulanek, D and Varga, Z and Rákosy, L and Schmitt, T}, title = {Pronounced mito-nuclear discordance and various Wolbachia infections in the water ringlet Erebia pronoe have resulted in a complex phylogeographic structure.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {5175}, pmid = {35338196}, issn = {2045-2322}, mesh = {Animals ; *Butterflies/genetics ; DNA, Mitochondrial/genetics ; Genetic Variation ; *MELAS Syndrome ; Male ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Several morphological and mitochondrial lineages of the alpine ringlet butterfly species Erebia pronoe have been described, indicating a complex phylogenetic structure. However, the existing data were insufficient and allow neither a reconstruction of the biogeographic history, nor an assessment of the genetic lineages. Therefore, we analysed mitochondrial (COI, NDI) and nuclear (EF1α, RPS5) gene sequences and compared them with sequences from the sister species Erebia melas. Additionally, we combined this information with morphometric data of the male genitalia and the infection patterns with Wolbachia strains, based on a WSP analysis. We obtained a distinct phylogeographic structure within the E. pronoe-melas complex with eight well-distinguishable geographic groups, but also a remarkable mito-nuclear discordance. The mito-nuclear discordance in E. melas and E. pronoe glottis can be explained by different ages of Wolbachia infections with different Wolbachia strains, associated selective sweeps, and hybridisation inhibition. Additionally, we found indications for incipient speciation of E. pronoe glottis in the Pyrenees and a pronounced range dynamic within and among the other high mountain systems of Europe. Our results emphasize the importance of combined approaches in reconstructing biogeographic patterns and evaluating phylogeographic splits.}, } @article {pmid35338130, year = {2022}, author = {Wang, H and Xiao, Y and Chen, X and Zhang, M and Sun, G and Wang, F and Wang, L and Zhang, H and Zhang, X and Yang, X and Li, W and Wei, Y and Yao, D and Zhang, B and Li, J and Cui, W and Wang, F and Chen, C and Shen, W and Su, D and Bai, F and Huang, J and Ye, S and Zhang, L and Ji, X and Wang, W and Wang, Z and Hochstrasser, M and Yang, H}, title = {Crystal Structures of Wolbachia CidA and CidB Reveal Determinants of Bacteria-induced Cytoplasmic Incompatibility and Rescue.}, journal = {Nature communications}, volume = {13}, number = {1}, pages = {1608}, pmid = {35338130}, issn = {2041-1723}, support = {R35 GM136325/GM/NIGMS NIH HHS/United States ; R37 GM046904/GM/NIGMS NIH HHS/United States ; R01 GM046904/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/genetics ; Cytosol ; Drosophila melanogaster/genetics ; Female ; Male ; Saccharomyces cerevisiae ; Symbiosis/genetics ; *Wolbachia/genetics/metabolism ; }, abstract = {Cytoplasmic incompatibility (CI) results when Wolbachia bacteria-infected male insects mate with uninfected females, leading to embryonic lethality. "Rescue" of viability occurs if the female harbors the same Wolbachia strain. CI is caused by linked pairs of Wolbachia genes called CI factors (CifA and CifB). The co-evolution of CifA-CifB pairs may account in part for the incompatibility patterns documented in insects infected with different Wolbachia strains, but the molecular mechanisms remain elusive. Here, we use X-ray crystallography and AlphaFold to analyze the CI factors from Wolbachia strain wMel called CidA[wMel] and CidB[wMel]. Substituting CidA[wMel] interface residues with those from CidA[wPip] (from strain wPip) enables the mutant protein to bind CidB[wPip] and rescue CidB[wPip]-induced yeast growth defects, supporting the importance of CifA-CifB interaction in CI rescue. Sequence divergence in CidA[wPip] and CidB[wPip] proteins affects their pairwise interactions, which may help explain the complex incompatibility patterns of mosquitoes infected with different wPip strains.}, } @article {pmid35336091, year = {2022}, author = {Petrone, JR and Muñoz-Beristain, A and Glusberger, PR and Russell, JT and Triplett, EW}, title = {Unamplified, Long-Read Metagenomic Sequencing Approach to Close Endosymbiont Genomes of Low-Biomass Insect Populations.}, journal = {Microorganisms}, volume = {10}, number = {3}, pages = {}, pmid = {35336091}, issn = {2076-2607}, support = {CRDF: 60100000-209- 2200-CRRNTG000780-GRANTP0004552-1//CRDF/ ; }, abstract = {With the current advancements in DNA sequencing technology, the limiting factor in long-read metagenomic assemblies is now the quantity and quality of input DNA. Although these requirements can be met through the use of axenic bacterial cultures or large amounts of biological material, insect systems that contain unculturable bacteria or that contain a low amount of available DNA cannot fully utilize the benefits of third-generation sequencing. The citrus greening disease insect vector Diaphorina citri is an example that exhibits both of these limitations. Although endosymbiont genomes have mostly been closed after the short-read sequencing of amplified template DNA, creating de novo long-read genomes from the unamplified DNA of an insect population may benefit communities using bioinformatics to study insect pathosystems. Here all four genomes of the infected D. citri microbiome were sequenced to closure using unamplified template DNA and two long-read sequencing technologies. Avoiding amplification bias and using long reads to assemble the bacterial genomes allowed for the circularization of the Wolbachia endosymbiont of Diaphorina citri for the first time and paralleled the annotation context of all four reference genomes without utilizing a traditional hybrid assembly. The strategies detailed here are suitable for the sequencing of other insect systems for which the input DNA, time, and cost are an issue.}, } @article {pmid35335697, year = {2022}, author = {Allman, MJ and Slack, AJ and Abello, NP and Lin, YH and O'Neill, SL and Robinson, AJ and Flores, HA and Joubert, DA}, title = {Trash to Treasure: How Insect Protein and Waste Containers Can Improve the Environmental Footprint of Mosquito Egg Releases.}, journal = {Pathogens (Basel, Switzerland)}, volume = {11}, number = {3}, pages = {}, pmid = {35335697}, issn = {2076-0817}, support = {na//Australian Government Research Training Program (RTP) Scholarship/ ; na//Institute of Vector-borne Disease Scholarship/ ; Strategic 102591/Z/13/Z//Wellcome Trust Fund/ ; OPP 1153619//Bill and Melinda Gates Foundation/ ; }, abstract = {Release and subsequent establishment of Wolbachia-infected Aedes aegypti in native mosquito populations has successfully reduced mosquito-borne disease incidence. While this is promising, further development is required to ensure that this method is scalable and sustainable. Egg release is a beneficial technique that requires reduced onsite resources and increases community acceptance; however, its incidental ecological impacts must be considered to ensure sustainability. In this study, we tested a more environmentally friendly mosquito rearing and release approach through the encapsulation of diet and egg mixtures and the subsequent utilization of waste containers to hatch and release mosquitoes. An ecologically friendly diet mix was specifically developed and tested for use in capsules, and we demonstrated that using either cricket or black soldier fly meal as a substitute for beef liver powder had no adverse effects on fitness or Wolbachia density. We further encapsulated both the egg and diet mixes and demonstrated no loss in viability. To address the potential of increased waste generation through disposable mosquito release containers, we tested reusing commonly found waste containers (aluminum and tin cans, PET, and glass bottles) as an alternative, conducting a case study in Kiribati to assess the concept's cultural, political, and economic applicability. Our results showed that mosquito emergence and fitness was maintained with a variety of containers, including when tested in the field, compared to control containers, and that there are opportunities to implement this method in the Pacific Islands in a way that is culturally considerate and cost-effective.}, } @article {pmid35323577, year = {2022}, author = {Kaczmarczyk-Ziemba, A and Kucharczyk, H and Kucharczyk, M and Kucharska, K}, title = {Integrative Insight into Relationships between Florivorous Thrips Haplothrips leucanthemi and H. niger (Insecta, Thysanoptera).}, journal = {Insects}, volume = {13}, number = {3}, pages = {}, pmid = {35323577}, issn = {2075-4450}, abstract = {Haplothrips niger is recognized as a parthenogenetic form of H. leucanthemi and is also considered to be a pest in clover-seed plantations. On the contrary, some researchers highlight the distinctiveness of H. niger and H. leucanthemi. Taking into account these two points of view, as well as the lack of molecular studies investigating the relationship between the mentioned thrips, we decided to perform analyses of both mitochondrial (COI) and nuclear markers (28S and ITS2) to determine the genetic diversity of H. leucanthemi and H. niger. Additionally, as a part of an integrative approach, we determined and analyzed their microbiota profiles, based on high-throughput 16S rRNA gene sequencing. The results of the molecular analyses revealed high intraspecific diversity of H. leucanthemi and did not support the distinctiveness of H. niger. The identified microbiota profiles were similar in both species and the performed analyses also did not support the distinctiveness of H. niger. Significant differences were, in turn, observed between H. leucanthemi and H. niger larvae. Moreover, two known endosymbiotic bacteria were found in the analyzed microbiota profiles (i.e., Wolbachia and Rickettsia). Nevertheless, these symbionts were not predominantly found in the bacterial communities that are associated with H. niger and thus, its impact on the parthenogenetic mode of its reproduction seems less likely.}, } @article {pmid35319146, year = {2022}, author = {Gu, X and Ross, PA and Rodriguez-Andres, J and Robinson, KL and Yang, Q and Lau, MJ and Hoffmann, AA}, title = {A wMel Wolbachia variant in Aedes aegypti from field-collected Drosophila melanogaster with increased phenotypic stability under heat stress.}, journal = {Environmental microbiology}, volume = {24}, number = {4}, pages = {2119-2135}, pmid = {35319146}, issn = {1462-2920}, mesh = {*Aedes/genetics ; Animals ; Drosophila melanogaster/genetics ; Heat-Shock Response ; Mosquito Vectors ; *Wolbachia/genetics ; }, abstract = {Mosquito-borne diseases remain a major cause of morbidity and mortality. Population replacement strategies involving the wMel strain of Wolbachia are being used widely to control mosquito-borne diseases. However, these strategies may be influenced by temperature because wMel is vulnerable to heat. wMel infections in Drosophila melanogaster are genetically diverse, but few transinfections of wMel variants have been generated in Aedes aegypti. Here, we successfully transferred a wMel variant (termed wMelM) originating from a field-collected D. melanogaster into Ae. aegypti. The new wMelM variant (clade I) is genetically distinct from the original wMel transinfection (clade III), and there are no genomic differences between wMelM in its original and transinfected host. We compared wMelM with wMel in its effects on host fitness, temperature tolerance, Wolbachia density, vector competence, cytoplasmic incompatibility and maternal transmission under heat stress in a controlled background. wMelM showed a higher heat tolerance than wMel, likely due to higher overall densities within the mosquito. Both wMel variants had minimal host fitness costs, complete cytoplasmic incompatibility and maternal transmission, and dengue virus blocking under laboratory conditions. Our results highlight phenotypic differences between Wolbachia variants and wMelM shows potential as an alternative strain in areas with strong seasonal temperature fluctuations.}, } @article {pmid35318403, year = {2022}, author = {Soh, LS and Veera Singham, G}, title = {Bacterial symbionts influence host susceptibility to fenitrothion and imidacloprid in the obligate hematophagous bed bug, Cimex hemipterus.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {4919}, pmid = {35318403}, issn = {2045-2322}, mesh = {Animals ; *Bedbugs/physiology ; *Ectoparasitic Infestations ; Fenitrothion ; Humans ; Insecticide Resistance ; *Insecticides/pharmacology ; Neonicotinoids ; Nitro Compounds ; *Pyrethrins/pharmacology ; Rifampin/pharmacology ; }, abstract = {The use of insecticides remains important in managing pest insects. Over the years, many insects manifested physiological and behavioral modifications resulting in reduced efficacy of insecticides targeted against them. Emerging evidence suggests that bacterial symbionts could modulate susceptibility of host insects against insecticides. Here, we explore the influence of host microbiota in affecting the susceptibility of insect host against different insecticides in the blood-sucking bed bug, Cimex hemipterus. Rifampicin antibiotic treatment resulted in increased susceptibility to fenitrothion and imidacloprid, but not against deltamethrin. Meanwhile, the host fitness parameters measured in the present study were not significantly affected by rifampicin treatment, suggesting the role of bacterial symbionts influencing susceptibility against the insecticides. 16S metagenomics sequencing revealed a drastic shift in the composition of several bacterial taxa following rifampicin treatment. The highly abundant Alphaproteobacteria (Wolbachia > 90%) and Gammaproteobacteria (Yersinia > 6%) in control bed bugs were significantly suppressed and replaced by Actinobacteria, Bacilli, and Betaproteobacteria in the rifampicin treated F1 bed bugs, suggesting possibilities of Wolbachia mediating insecticide susceptibility in C. hemipterus. However, no significant changes in the total esterase, GST, and P450 activities were observed following rifampicin treatment, indicating yet unknown bacterial mechanisms explaining the observed phenomena. Re-inoculation of microbial content from control individuals regained the tolerance of rifampicin treated bed bugs to imidacloprid and fenitrothion. This study provides a foundation for a symbiont-mediated mechanism in influencing insecticide susceptibility that was previously unknown to bed bugs.}, } @article {pmid35314082, year = {2022}, author = {Wang, GH and Du, J and Chu, CY and Madhav, M and Hughes, GL and Champer, J}, title = {Symbionts and gene drive: two strategies to combat vector-borne disease.}, journal = {Trends in genetics : TIG}, volume = {38}, number = {7}, pages = {708-723}, doi = {10.1016/j.tig.2022.02.013}, pmid = {35314082}, issn = {0168-9525}, support = {NIHR2000907/DH_/Department of Health/United Kingdom ; BB/T001240/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; V011278/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; *Culicidae/genetics ; *Gene Drive Technology ; *Malaria/genetics/prevention & control ; Mosquito Vectors/genetics ; *Wolbachia/genetics ; }, abstract = {Mosquitoes bring global health problems by transmitting parasites and viruses such as malaria and dengue. Unfortunately, current insecticide-based control strategies are only moderately effective because of high cost and resistance. Thus, scalable, sustainable, and cost-effective strategies are needed for mosquito-borne disease control. Symbiont-based and genome engineering-based approaches provide new tools that show promise for meeting these criteria, enabling modification or suppression approaches. Symbiotic bacteria like Wolbachia are maternally inherited and manipulate mosquito host reproduction to enhance their vertical transmission. Genome engineering-based gene drive methods, in which mosquitoes are genetically altered to spread drive alleles throughout wild populations, are also proving to be a potentially powerful approach in the laboratory. Here, we review the latest developments in both symbionts and gene drive-based methods. We describe some notable similarities, as well as distinctions and obstacles, relating to these promising technologies.}, } @article {pmid35303931, year = {2022}, author = {Hosseini, SH and Manshori-Ghaishghorshagh, F and Ramezani, M and Nayebzadeh, H and Ahoo, MB and Eslamian, A and Soltani, M and Jamshidi, S and Bezerra-Santos, MA and Jalousian, F and Sazmand, A and Otranto, D}, title = {Canine microfilaraemia in some regions of Iran.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {90}, pmid = {35303931}, issn = {1756-3305}, support = {Fateme Manshori-Ghaishghorshagh//University of Tehran/ ; Ahdieh Eslamian Theses//University of Tehran/ ; Mohammad Ramezani thesis//Lorestan University/ ; }, mesh = {Animals ; *Dirofilaria immitis/genetics ; *Dirofilaria repens/genetics ; *Dog Diseases/parasitology ; Dogs ; Iran/epidemiology ; Phylogeny ; }, abstract = {BACKGROUND: Dirofilaria immitis and Dirofilaria repens are vector-borne zoonotic parasites which affect mainly dogs and humans worldwide. In Iran, information about the distribution of those nematodes is scant in several regions. Therefore, we investigated the prevalence of these filarial parasites in stray dogs from five Iranian provinces where no information about these parasites is available.

METHODS: Blood samples were collected from 344 stray dogs in five provinces of Iran (i.e. Mazandaran, Gilan, Esfahan, Qazvin and Loresan). The presence of microfilariae was assessed using direct smear, modified Knott's test, molecular detection of filarial DNA (cox1 gene) and Wolbachia endosymbiont of parasitic nematodes (ftsZ gene) by conventional PCR (cPCR). All of the PCR products were sequenced and phylogenetic analysis was performed.

RESULTS: In total, 75 dogs (21.8%) were found to be positive for D. immitis by cPCR. Infection was detected in all provinces, with the highest prevalence in Gilan province (22/28; 78.6%). Acanthocheilonema reconditum was diagnosed in five dogs (1.4%) from three provinces (i.e. Esfahan, Mazandaran, Gilan). Two dogs were infected with both parasites and three were only infected with A. reconditum. Dirofilaria repens infection was not found in the examined population. Representative sequences of the D. immitis cox1 gene from dogs from the northern provinces (Mazandaran, Gilan, Qazvin) were grouped together and distinctly separate from the ones from western and central provinces (Lorestan and Esfahan), suggesting that different nematode populations are present in the country.

CONCLUSION: The data reported herein fill existing gaps in knowledge about canine filarial infection in two Iranian provinces and record the highest prevalence of D. immitis ever reported in the country (i.e. 78.6%). A geographical review of the literature about Dirofilaria spp. and A. reconditum infections in dogs and humans has also been summarized, indicating that D. immitis and D. repens are distributed in 22 of 31 provinces in Iran, whereas A. reconditum is present in fewer regions. Effective control strategies are advocated for owned dogs, and a national program for the management of stray dogs is needed to minimize the risk of infection in animals and humans.}, } @article {pmid35294495, year = {2022}, author = {Bhattacharya, T and Yan, L and Crawford, JM and Zaher, H and Newton, ILG and Hardy, RW}, title = {Differential viral RNA methylation contributes to pathogen blocking in Wolbachia-colonized arthropods.}, journal = {PLoS pathogens}, volume = {18}, number = {3}, pages = {e1010393}, pmid = {35294495}, issn = {1553-7374}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; R01 GM112641/GM/NIGMS NIH HHS/United States ; R01 GM141474/GM/NIGMS NIH HHS/United States ; R21 AI153785/AI/NIAID NIH HHS/United States ; }, mesh = {5-Methylcytosine/metabolism ; *Aedes ; *Alphavirus/genetics ; Animals ; *Arthropods/genetics ; *Flavivirus/genetics ; Methylation ; Methyltransferases/genetics/metabolism ; RNA, Viral/genetics/metabolism ; Virus Replication ; *Wolbachia/physiology ; }, abstract = {Arthropod endosymbiont Wolbachia pipientis is part of a global biocontrol strategy to reduce the replication of mosquito-borne RNA viruses such as alphaviruses. We previously demonstrated the importance of a host cytosine methyltransferase, DNMT2, in Drosophila and viral RNA as a cellular target during pathogen-blocking. Here we report a role for DNMT2 in Wolbachia-induced alphavirus inhibition in Aedes species. Expression of DNMT2 in mosquito tissues, including the salivary glands, is elevated upon virus infection. Notably, this is suppressed in Wolbachia-colonized animals, coincident with reduced virus replication and decreased infectivity of progeny virus. Ectopic expression of DNMT2 in cultured Aedes cells is proviral, increasing progeny virus infectivity, and this effect of DNMT2 on virus replication and infectivity is dependent on its methyltransferase activity. Finally, examining the effects of Wolbachia on modifications of viral RNA by LC-MS show a decrease in the amount of 5-methylcytosine modification consistent with the down-regulation of DNMT2 in Wolbachia colonized mosquito cells and animals. Collectively, our findings support the conclusion that disruption of 5-methylcytosine modification of viral RNA is a vital mechanism operative in pathogen blocking. These data also emphasize the essential role of epitranscriptomic modifications in regulating fundamental alphavirus replication and transmission processes.}, } @article {pmid35292086, year = {2022}, author = {Weck, BC and Serpa, MCA and Ramos, VN and Luz, HR and Costa, FB and Ramirez, DG and Benatti, HR and Piovezan, U and Szabó, MPJ and Marcili, A and Krawczak, FS and Muñoz-Leal, S and Labruna, MB}, title = {Novel genotypes of Hepatozoon spp. in small mammals, Brazil.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {87}, pmid = {35292086}, issn = {1756-3305}, support = {2013/18046-7//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2017/04249-4//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; }, mesh = {Animals ; Brazil/epidemiology ; *Carnivora ; *Eucoccidiida/genetics ; Haplotypes ; Phylogeny ; }, abstract = {BACKGROUND: Small mammals (rodents and marsupials) have been poorly explored for the occurrence of apicomplexan (genus Hepatozoon and genera of the order Piroplasmorida) and Anaplasmataceae agents in Brazil. Thus, this study investigated the occurrence of Hepatozoon spp., Piroplasmorida, and Anaplasmataceae agents in small mammals in seven forest fragments in Brazil.

METHODS: During 2015-2018, small mammals were captured in six forest fragments in the State of São Paulo (Cerrado and Atlantic Forest biomes) and one fragment in the State of Mato Grosso do Sul (Pantanal biome). Mammal blood, liver, spleen, and lung samples were tested molecularly for the presence of DNA of Hepatozoon, Piroplasmorida, and Anaplasmataceae agents.

RESULTS: A total of 524 mammals were captured, comprising seven species of marsupials, 14 rodents, two carnivores, and one Cingulata. Four novel haplotypes (1, 2, 3, 4) of Hepatozoon spp. were detected in small mammals from different biomes. In São Paulo state, haplotype 1 was detected in rodents from Cerrado and a transition area of Cerrado and Atlantic Forest biomes, whereas haplotype 2 was detected in rodents from the Atlantic Forest biome. On the other hand, haplotypes 3 and 4 were restricted to rodents and marsupials, respectively, from the Pantanal biome of Mato Grosso do Sul. No host species shared more than one haplotype. Despite these distinct geographical and host associations, our phylogenetic analyses indicated that the four Hepatozoon haplotypes belonged to the same clade that contained nearly all haplotypes previously reported on rodents and marsupials, in addition to several reptile-associated haplotypes from different parts of the world. No mammal samples yielded detectable DNA of Piroplasmorida agents. On the other hand, the Anaplasmataceae-targeted polymerase chain reaction (PCR) assay amplified a sequence 100% identical to the Wolbachia pipientis endosymbiont of the rodent filarid Litomosoides galizai.

CONCLUSIONS: We report a variety of Hepatozoon haplotypes associated with small mammals in three Brazilian biomes: Cerrado, Atlantic Forest, and Pantanal. Through phylogenetic analyses, the Hepatozoon agents grouped in the rodent-marsupial-reptile large clade of Hepatozoon spp. from the world. The detection of a W. pipientis associated with the rodent filarid L. galizai indicates that the rodent was infected by filarial nematodes.}, } @article {pmid35286393, year = {2022}, author = {Ndiaye, EHI and Diatta, G and Diarra, AZ and Berenger, JM and Bassene, H and Mediannikov, O and Bouganali, C and Sokhna, C and Parola, P}, title = {Morphological, Molecular and MALDI-TOF MS Identification of Bedbugs and Associated Wolbachia Species in Rural Senegal.}, journal = {Journal of medical entomology}, volume = {59}, number = {3}, pages = {1019-1032}, doi = {10.1093/jme/tjac019}, pmid = {35286393}, issn = {1938-2928}, mesh = {Animals ; *Bedbugs/anatomy & histology ; *Ectoparasitic Infestations ; Senegal ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; *Wolbachia ; }, abstract = {Bed bugs are known to carry several microorganisms. The purpose of this study was to assess the prevalence of bed bug infestation in two rural areas of Senegal and determine the species present in the population. A screening was conducted to detect some arthropod associated pathogenic bacteria in bed bugs and to evaluate the prevalence of endosymbiont carriage. One survey took place in 17 villages in Niakhar and two surveys in Dielmo and Ndiop and surroundings area in the same 20 villages. Bed bugs collected were identified morphologically and by MALDI-TOF MS tools. Microorganisms screening was performed by qPCR and confirmed by sequencing. During the survey in the Niakhar region, only one household 1/255 (0.4%) in the village of Ngayokhem was found infested by bed bugs. In a monitoring survey of the surroundings of Dielmo and Ndiop area, high prevalence was found during the two rounds of surveys in 65/314 (21%) in 16/20 villages (January-March) and 93/351 (26%) in 19/20 villages (December). All bed bugs were morphologically identified as the species Cimex hemipterus, of which 285/1,637 (17%) were randomly selected for MALDI-TOF MS analysis and bacteria screening. Among the Bacteria tested only Wolbachia (Alphaproteobacteria, Rickettsiales, Rickettsiaceae) DNA was found in 248/276 (90%) of the bedbugs. We briefly describe a high level of non-generalized bed bug infestation in rural Senegal and the diversity of Wolbachia strains carried by C. hemipterus. This study opens perspectives for raising household awareness of bed bug infestations and possibilities for appropriate control.}, } @article {pmid35271765, year = {2022}, author = {Mao, B and Zhang, W and Zheng, Y and Li, D and Chen, MY and Wang, YF}, title = {Comparative phosphoproteomics reveal new candidates in the regulation of spermatogenesis of Drosophila melanogaster.}, journal = {Insect science}, volume = {29}, number = {6}, pages = {1703-1720}, doi = {10.1111/1744-7917.13031}, pmid = {35271765}, issn = {1744-7917}, support = {31672352//National Natural Science Foundation of China/ ; 31872288//National Natural Science Foundation of China/ ; }, mesh = {Female ; Male ; Animals ; *Drosophila melanogaster/genetics ; Proteomics ; Semen ; Spermatogenesis ; *Wolbachia/physiology ; Phosphoproteins ; }, abstract = {The most common phenotype induced by the endosymbiont Wolbachia in insects is cytoplasmic incompatibility, where none or fewer progenies can be produced when Wolbachia-infected males mate with uninfected females. This suggests that some modifications are induced in host sperms during spermatogenesis by Wolbachia. To identify the proteins whose phosphorylation states play essential roles in male reproduction in Drosophila melanogaster, we applied isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic strategy combined with titanium dioxide (TiO2) enrichment to compare the phosphoproteome of Wolbachia-infected with that of uninfected male reproductive systems in D. melanogaster. We identified 182 phosphopeptides, defining 140 phosphoproteins, that have at least a 1.2 fold change in abundance with a P-value of <0.05. Most of the differentially abundant phosphoproteins (DAPPs) were associated with microtubule cytoskeleton organization and spermatid differentiation. The DAPPs included proteins already known to be associated with spermatogenesis, as well as many not previously studied during this process. Six genes coding for DAPPs were knocked down, respectively, in Wolbachia-free fly testes. Among them, Slmap knockdown caused the most severe damage in spermatogenesis, with no mature sperm observed in seminal vesicles. Immunofluorescence staining showed that the formation of individualization complex composed of actin cones was completely disrupted. These results suggest that Wolbachia may induce wide changes in the abundance of phosphorylated proteins which are closely related to male reproduction. By identifying phospho-modulated proteins we also provide a significant candidate set for future studies on their roles in spermatogenesis.}, } @article {pmid35266572, year = {2022}, author = {Matthews, ML and Covey, HO and Drolet, BS and Brelsfoard, CL}, title = {Wolbachia wAlbB inhibits bluetongue and epizootic hemorrhagic fever viruses in Culicoides midge cells.}, journal = {Medical and veterinary entomology}, volume = {36}, number = {3}, pages = {320-328}, pmid = {35266572}, issn = {1365-2915}, mesh = {Animals ; *Bluetongue ; *Bluetongue virus/physiology ; *Ceratopogonidae/physiology ; *Dengue Virus/genetics ; Real-Time Polymerase Chain Reaction/veterinary ; Sheep ; *Sheep Diseases ; *Wolbachia/genetics ; }, abstract = {Culicoides midges are hematophagous insects that transmit arboviruses of veterinary importance. These viruses include bluetongue virus (BTV) and epizootic hemorrhagic fever virus (EHDV). The endosymbiont Wolbachia pipientis Hertig spreads rapidly through insect host populations and has been demonstrated to inhibit viral pathogen transmission in multiple mosquito vectors. Here, we have demonstrated a replication inhibitory effect on BTV and EHDV in a Wolbachia (wAlbB strain)-infected Culicoides sonorensis Wirth and Jones W8 cell line. Viral replication was significantly reduced by day 5 for BTV and by day 2 for EHDV as detected by real-time polymerase chain reaction (RT-qPCR) of the non-structural NS3 gene of both viruses. Evaluation of innate cellular immune responses as a cause of the inhibitory effect showed responses associated with BTV but not with EHDV infection. Wolbachia density also did not play a role in the observed pathogen inhibitory effects, and an alternative hypothesis is suggested. Applications of Wolbachia-mediated pathogen interference to impact disease transmission by Culicoides midges are discussed.}, } @article {pmid35264574, year = {2022}, author = {Klimov, PB and Chetverikov, PE and Dodueva, IE and Vishnyakov, AE and Bolton, SJ and Paponova, SS and Lutova, LA and Tolstikov, AV}, title = {Symbiotic bacteria of the gall-inducing mite Fragariocoptes setiger (Eriophyoidea) and phylogenomic resolution of the eriophyoid position among Acari.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {3811}, pmid = {35264574}, issn = {2045-2322}, mesh = {Animals ; Bacteria ; Biological Evolution ; In Situ Hybridization, Fluorescence ; *Mites/genetics ; Phylogeny ; Plants ; }, abstract = {Eriophyoid mites represent a hyperdiverse, phytophagous lineage with an unclear phylogenetic position. These mites have succeeded in colonizing nearly every seed plant species, and this evolutionary success was in part due to the mites' ability to induce galls in plants. A gall is a unique niche that provides the inducer of this modification with vital resources. The exact mechanism of gall formation is still not understood, even as to whether it is endogenic (mites directly cause galls) or exogenic (symbiotic microorganisms are involved). Here we (i) investigate the phylogenetic affinities of eriophyoids and (ii) use comparative metagenomics to test the hypothesis that the endosymbionts of eriophyoid mites are involved in gall formation. Our phylogenomic analysis robustly inferred eriophyoids as closely related to Nematalycidae, a group of deep-soil mites belonging to Endeostigmata. Our comparative metagenomics, fluorescence in situ hybridization, and electron microscopy experiments identified two candidate endosymbiotic bacteria shared across samples, however, it is unlikely that they are gall inducers (morphotype1: novel Wolbachia, morphotype2: possibly Agrobacterium tumefaciens). We also detected an array of plant pathogens associated with galls that may be vectored by the mites, and we determined a mite pathogenic virus (Betabaculovirus) that could be tested for using in biocontrol of agricultural pest mites.}, } @article {pmid35256789, year = {2022}, author = {Adams, KL and Abernathy, DG and Willett, BC and Selland, EK and Itoe, MA and Catteruccia, F}, title = {Author Correction: Wolbachia cifB induces cytoplasmic incompatibility in the malaria mosquito vector.}, journal = {Nature microbiology}, volume = {7}, number = {4}, pages = {600}, doi = {10.1038/s41564-022-01098-9}, pmid = {35256789}, issn = {2058-5276}, support = {PGSD3-488065-2016//Gouvernement du Canada | Natural Sciences and Engineering Research Council of Canada (Conseil de Recherches en Sciences Naturelles et en Génie du Canada)/ ; OPP1174120//Bill and Melinda Gates Foundation (Bill & Melinda Gates Foundation)/ ; }, } @article {pmid35252033, year = {2022}, author = {Yu, S and Wang, J and Luo, X and Zheng, H and Wang, L and Yang, X and Wang, Y}, title = {Transmission-Blocking Strategies Against Malaria Parasites During Their Mosquito Stages.}, journal = {Frontiers in cellular and infection microbiology}, volume = {12}, number = {}, pages = {820650}, pmid = {35252033}, issn = {2235-2988}, mesh = {Animals ; *Culicidae ; Humans ; *Malaria/prevention & control ; *Malaria, Falciparum/parasitology ; *Parasites ; Plasmodium falciparum ; }, abstract = {Malaria is still the most widespread parasitic disease and causes the most infections globally. Owing to improvements in sanitary conditions and various intervention measures, including the use of antimalarial drugs, the malaria epidemic in many regions of the world has improved significantly in the past 10 years. However, people living in certain underdeveloped areas are still under threat. Even in some well-controlled areas, the decline in malaria infection rates has stagnated or the rates have rebounded because of the emergence and spread of drug-resistant malaria parasites. Thus, new malaria control methods must be developed. As the spread of the Plasmodium parasite is dependent on the part of its life cycle that occurs in mosquitoes, to eliminate the possibility of malaria infections, transmission-blocking strategies against the mosquito stage should be the first choice. In fact, after the gametocyte enters the mosquito body, it undergoes a series of transformation processes over a short period, thus providing numerous potential blocking targets. Many research groups have carried out studies based on targeting the blocking of transmission during the mosquito phase and have achieved excellent results. Meanwhile, the direct killing of mosquitoes could also significantly reduce the probability of malaria infections. Microorganisms that display complex interactions with Plasmodium, such as Wolbachia and gut flora, have shown observable transmission-blocking potential. These could be used as a biological control strategy and play an important part in blocking the transmission of malaria.}, } @article {pmid35247466, year = {2022}, author = {Bojko, J and McCoy, KA and Blakeslee, AMH}, title = {'Candidatus Mellornella promiscua' n. gen. n. sp. (Alphaproteobacteria: Rickettsiales: Anaplasmataceae): An intracytoplasmic, hepatopancreatic, pathogen of the flatback mud crab, Eurypanopeus depressus.}, journal = {Journal of invertebrate pathology}, volume = {190}, number = {}, pages = {107737}, doi = {10.1016/j.jip.2022.107737}, pmid = {35247466}, issn = {1096-0805}, mesh = {*Alphaproteobacteria/genetics ; *Anaplasmataceae/genetics ; Animals ; *Brachyura/genetics ; Ecosystem ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsiales/genetics ; }, abstract = {Bacterial pathogens are a long-standing threat to the longevity and survival of crustacean hosts. Their presence and continuing emergence require close monitoring to understand their impact on fished, cultured, and wild crustacean populations. We describe a new bacterial pathogen belonging to the Anaplasmataceae family (Alphaproteobacteria: Rickettsiales), providing pathological, ultrastructural, phylogenetic, and genomic evidence to determine a candidate genus and species ('Candidatus Mellornella promiscua'). This bacterium was found to infect the mud crab, Eurypanopeus depressus, on the North Carolina coastline (USA) at a prevalence of 10.8%. 'Candidatus Mellornella promiscua' was often observed in co-infection with the rhizocephalan barnacle, Loxothylacus panopaei. The bacterium was only found in the hepatopancreas of the mud crab host, causing cytoplasmic hypertrophy, tubule necrosis, large plaques within the cytoplasm of the host cell, and an abundance of sex-pili. The circular genome of the bacterium is 1,013,119 bp and encodes 939 genes in total. Phylogenetically, the new bacterium branches within the Anaplasmataceae. The genome is dissimilar from other described bacteria, with 16S gene similarity observed at a maximum of 85.3% to a Wolbachia endosymbiont. We explore this novel bacterial pathogen using genomic, phylogenetic, ultrastructural, and pathological methods, discussing these results in light of current bacterial taxonomy, similarity to other bacterial pathogens, and the potential impact upon the surrounding disease ecology of the host and benthic ecosystem.}, } @article {pmid35237407, year = {2022}, author = {Vatandoost, H and Hanafi-Bojd, AA and Nikpoor, F and Raeisi, A and Abai, MR and Zaim, M}, title = {Situation of insecticide resistance in malaria vectors in the World Health Organization of Eastern Mediterranean region 1990-2020.}, journal = {Toxicology research}, volume = {11}, number = {1}, pages = {1-21}, pmid = {35237407}, issn = {2045-452X}, abstract = {Malaria is the most important mosquito-borne disease, which is transmitted by Anopheles species. According to the recent report of World Health Organization, there were an estimated 229 million malaria cases in 2019 in 87 malaria endemic countries. There are several vector control method specially using pesticides. The aim of this study was to collect all the data about insecticide resistant of malaria vectors in the Eastern Mediterranean region countries. In this study, all the published papers related to insecticide resistance of malaria vectors in the region were collected and resistant status of vectors in the countries was mapped. Resistance to different insecticide classes such as pyrethroids, organophosphate, organochlorine and carbamates were evaluated using World Health Organization guidelines. Results showed a wide variety of susceptibility/resistance status to these chemicals according to the location, historical context of pesticide used, genetic background of vectors, age and abdominal conditions of adults may play a role in the susceptibility status of these species to different insecticides. The results of this study is providing a guideline for countries to manage their vector control activities against insecticide resistance of malaria vectors and provide novel approaches such as male sterility, using Wolbachia, using new insecticide with new mode of action.}, } @article {pmid35237241, year = {2021}, author = {Li, J and Wei, X and Huang, D and Xiao, J}, title = {The Phylosymbiosis Pattern Between the Fig Wasps of the Same Genus and Their Associated Microbiota.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {800190}, pmid = {35237241}, issn = {1664-302X}, abstract = {Microbial communities can be critical for many metazoans, which can lead to the observation of phylosymbiosis with phylogenetically related species sharing similar microbial communities. Most of the previous studies on phylosymbiosis were conducted across the host families or genera. However, it is unclear whether the phylosymbiosis signal is still prevalent at lower taxonomic levels. In this study, 54 individuals from six species of the fig wasp genus Ceratosolen (Hymenoptera: Agaonidae) collected from nine natural populations and their associated microbiota were investigated. The fig wasp species were morphologically identified and further determined by mitochondrial CO1 gene fragments and nuclear ITS2 sequences, and the V4 region of 16S rRNA gene was sequenced to analyze the bacterial communities. The results suggest a significant positive correlation between host genetic characteristics and microbial diversity characteristics, indicating the phylosymbiosis signal between the phylogeny of insect hosts and the associated microbiota in the lower classification level within a genus. Moreover, we found that the endosymbiotic Wolbachia carried by fig wasps led to a decrease in bacterial diversity of host-associated microbial communities. This study contributes to our understanding of the role of host phylogeny, as well as the role of endosymbionts in shaping the host-associated microbial community.}, } @article {pmid35236394, year = {2022}, author = {Collins, MH and Potter, GE and Hitchings, MDT and Butler, E and Wiles, M and Kennedy, JK and Pinto, SB and Teixeira, ABM and Casanovas-Massana, A and Rouphael, NG and Deye, GA and Simmons, CP and Moreira, LA and Nogueira, ML and Cummings, DAT and Ko, AI and Teixeira, MM and Edupuganti, S}, title = {EVITA Dengue: a cluster-randomized controlled trial to EValuate the efficacy of Wolbachia-InfecTed Aedes aegypti mosquitoes in reducing the incidence of Arboviral infection in Brazil.}, journal = {Trials}, volume = {23}, number = {1}, pages = {185}, pmid = {35236394}, issn = {1745-6215}, support = {UL1 TR001863/TR/NCATS NIH HHS/United States ; DMID17-0111//National Institute of Allergy and Infectious Diseases/ ; }, mesh = {*Aedes ; Animals ; Brazil/epidemiology ; Child ; *Dengue/epidemiology/prevention & control ; *Dengue Virus ; Humans ; Incidence ; Mosquito Vectors ; *Wolbachia ; *Zika Virus ; *Zika Virus Infection/epidemiology ; }, abstract = {BACKGROUND: Arboviruses transmitted by Aedes aegypti including dengue, Zika, and chikungunya are a major global health problem, with over 2.5 billion at risk for dengue alone. There are no licensed antivirals for these infections, and safe and effective vaccines are not yet widely available. Thus, prevention of arbovirus transmission by vector modification is a novel approach being pursued by multiple researchers. However, the field needs high-quality evidence derived from randomized, controlled trials upon which to base the implementation and maintenance of vector control programs. Here, we report the EVITA Dengue trial design (DMID 17-0111), which assesses the efficacy in decreasing arbovirus transmission of an innovative approach developed by the World Mosquito Program for vector modification of Aedes mosquitoes by Wolbachia pipientis.

METHODS: DMID 17-0111 is a cluster-randomized trial in Belo Horizonte, Brazil, with clusters defined by primary school catchment areas. Clusters (n = 58) will be randomized 1:1 to intervention (release of Wolbachia-infected Aedes aegypti mosquitoes) vs. control (no release). Standard vector control activities (i.e., insecticides and education campaigns for reduction of mosquito breeding sites) will continue as per current practice in the municipality. Participants (n = 3480, 60 per cluster) are children aged 6-11 years enrolled in the cluster-defining school and living within the cluster boundaries who will undergo annual serologic surveillance for arboviral infection. The primary objective is to compare sero-incidence of arboviral infection between arms.

DISCUSSION: DMID 17-0111 aims to determine the efficacy of Wolbachia-infected mosquito releases in reducing human infections by arboviruses transmitted by Aedes aegypti and will complement the mounting evidence for this method from large-scale field releases and ongoing trials. The trial also represents a critical step towards robustness and rigor for how vector control methods are assessed, including the simultaneous measurement and correlation of entomologic and epidemiologic outcomes. Data from this trial will inform further the development of novel vector control methods.

TRIAL REGISTRATION: ClinicalTrials.gov NCT04514107 . Registered on 17 August 2020 Primary sponsor: National Institute of Health, National Institute of Allergy and Infectious Diseases.}, } @article {pmid35231415, year = {2022}, author = {Stuckert, AMM and Matute, DR}, title = {Evolution: Environmental conditions determine how Wolbachia interacts with its host.}, journal = {Current biology : CB}, volume = {32}, number = {4}, pages = {R178-R180}, doi = {10.1016/j.cub.2022.01.010}, pmid = {35231415}, issn = {1879-0445}, mesh = {Animals ; *Arthropods ; Drosophila melanogaster/microbiology ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a genus of globally distributed endosymbiotic bacteria found in thousands of species of arthropods and nematodes. One strain, wMel, has spread globally but varies dramatically in frequency. In a new paper, Hague et al. discovered that the interaction of host genotype, symbiont genotype, and environment explains natural wMel variation.}, } @article {pmid35229443, year = {2022}, author = {Ashraf, HJ and Ramos Aguila, LC and Akutse, KS and Ilyas, M and Abbasi, A and Li, X and Wang, L}, title = {Comparative microbiome analysis of Diaphorina citri and its associated parasitoids Tamarixia radiata and Diaphorencyrtus aligarhensis reveals Wolbachia as a dominant endosymbiont.}, journal = {Environmental microbiology}, volume = {24}, number = {3}, pages = {1638-1652}, doi = {10.1111/1462-2920.15948}, pmid = {35229443}, issn = {1462-2920}, mesh = {Animals ; Bacteria ; *Citrus/microbiology ; *Hemiptera/microbiology ; *Microbiota ; *Wasps ; *Wolbachia ; }, abstract = {Microbiome analysis in a host-parasitoid interaction network was conducted to compare the taxonomic composition of bacterial communities of Diaphornia citri, Tamarixia radiata, and Diaphorencyrtus aligarhensis. The comparative analysis revealed differences in the composition and diversity of the symbiont populations across the host and its associated parasitoids. Proteobacteria was the most dominant phylum, representing 67.80% of the total bacterial community, while Candidatus Profftella armature and Wolbachia were the dominant genera across the host and parasitoids. There were clear differences observed in alpha and beta diversity of microbiota through the host and its associated parasitoids. The function prediction of bacterial communities and Pearson correlation analysis showed that specific bacterial communities displayed positive correlations with the carbohydrate metabolism pathway. Furthermore, when symbiotic bacteria were eliminated using a broad-spectrum antibiotic, tetracycline hydrochloride, the parasitoids' median survival time and longevity were significantly reduced. We confirmed the physiological effects of symbiotic bacteria on the fitness of parasitoids and demonstrated the effect of antibiotics in decreasing the food intake and measurement of amino acids in the hemolymph. This study sheds light on basic information about the mutualism between parasitoids and bacteria, which may be a potential source for biocontrol strategies for citrus psyllid, especially D. citri.}, } @article {pmid35209944, year = {2022}, author = {Moretti, R and Lampazzi, E and Damiani, C and Fabbri, G and Lombardi, G and Pioli, C and Desiderio, A and Serrao, A and Calvitti, M}, title = {Increased biting rate and decreased Wolbachia density in irradiated Aedes mosquitoes.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {67}, pmid = {35209944}, issn = {1756-3305}, mesh = {*Aedes/microbiology ; Animals ; Female ; In Situ Hybridization, Fluorescence ; *Infertility, Male ; Male ; Mosquito Control/methods ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Releasing considerable numbers of radiation-sterilized males is a promising strategy to suppress mosquito vectors. However, releases may also include small percentages of biting females, which translate to non-negligible numbers when releases are large. Currently, the effects of irradiation on host-seeking and host-biting behaviors have not been exhaustively investigated. Information is also lacking regarding the effects of sterilizing treatment on the endosymbiotic bacterium Wolbachia, which is known to affect the vector competence of infected mosquitos.

METHODS: To ascertain the effects of irradiation on females, the pupae of two Aedes albopictus strains, differing in their natural or artificial Wolbachia infection type, and Aedes aegypti-which is not infected by Wolbachia-were treated with various doses of X-rays and monitored for key fitness parameters and biting behavior over a period of 2 weeks. The effect of radiation on Wolbachia was investigated by quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FISH) analysis.

RESULTS: Partial Aedes albopictus female sterility was achieved at 28 Gy, but the number of weekly bites more than doubled compared to that of the controls. Radiation doses of 35 and 45 Gy completely inhibited progeny production but did not significantly affect the survival or flight ability of Ae. albopictus females and caused a tripling of the number of bites per female per week (compared to untreated controls). These results were also confirmed in Ae. aegypti after treatment at 50 Gy. Wolbachia density decreased significantly in 45-Gy-irradiated females, with the greatest decreases in the early irradiation group (26 ± 2-h-old pupae). Wolbachia density also decreased as adults aged. This trend was confirmed in ovaries but not in extra-ovarian tissues. FISH analysis showed a strongly reduced Wolbachia-specific fluorescence in the ovaries of 13 ± 1-day-old females.

CONCLUSIONS: These results suggest that, under sterile insect technique (SIT) programs, the vector capacity of a target population could increase with the frequency of the irradiated females co-released with the sterile males due to an increased biting rate. In the context of successful suppression, the related safety issues are expected to be generally negligible, but they should be conservatively evaluated when large-scale programs relying on imperfect sexing and high overflooding release ratios are run for long periods in areas endemic for arboviral diseases. Also, the effects of irradiation on the vector competence deserve further investigation.}, } @article {pmid35206789, year = {2022}, author = {Damiani, C and Cappelli, A and Comandatore, F and Montarsi, F and Serrao, A and Michelutti, A and Bertola, M and Mancini, MV and Ricci, I and Bandi, C and Favia, G}, title = {Wolbachia in Aedes koreicus: Rare Detections and Possible Implications.}, journal = {Insects}, volume = {13}, number = {2}, pages = {}, pmid = {35206789}, issn = {2075-4450}, support = {Far 2019//University of Camerino/ ; }, abstract = {The emerging distribution of new alien mosquito species was recently described in Europe. In addition to the invasion of Aedes albopictus, several studies have focused on monitoring and controlling other invasive Aedes species, as Aedes koreicus and Aedes japonicus. Considering the increasing development of insecticide resistance in Aedes mosquitoes, new control strategies, including the use of bacterial host symbionts, are proposed. However, little is known about the bacterial communities associated with these species, thus the identification of possible candidates for Symbiotic Control is currently limited. The characterization of the natural microbiota of field-collected Ae. koreicus mosquitoes from North-East Italy through PCR screening, identified native infections of Wolbachia in this species that is also largely colonized by Asaia bacteria. Since Asaia and Wolbachia are proposed as novel tools for Symbiotic Control, our study supports their use for innovative control strategies against new invasive species. Although the presence of Asaia was previously characterized in Ae. koreicus, our study characterized this Wolbachia strain, also inferring its phylogenetic position. The co-presence of Wolbachia and Asaia may provide additional information about microbial competition in mosquito, and to select suitable phenotypes for the suppression of pathogen transmission and for the manipulation of host reproduction in Ae. koreicus.}, } @article {pmid35206699, year = {2022}, author = {Ilinsky, Y and Lapshina, V and Verzhutsky, D and Fedorova, Y and Medvedev, S}, title = {Genetic Evidence of an Isolation Barrier between Flea Subspecies of Citellophilus tesquorum (Wagner, 1898) (Siphonaptera: Ceratophyllidae).}, journal = {Insects}, volume = {13}, number = {2}, pages = {}, pmid = {35206699}, issn = {2075-4450}, support = {19-04-00759//Russian Foundation for Basic Research/ ; 121031800061-7//Ministry of Science and Higher Education of the Russian Federation/ ; АААА-А19-119020790133-6//Ministry of Science and Higher Education of the Russian Federation/ ; }, abstract = {This study investigated the relationship between two subspecies of the Citellophilus tesquorum flea, C. t. altaicus and C. t. sungaris, which are vectors of the bacterium Yersinia pestis that causes human plague across the vast territories of the Palearctic. Adult fleas were collected from 16 localities and 11 populations in 2019 and 2020. Specimens were morphologically verified for subspecies status and analysed for mitochondrial cytochrome c oxidase subunit I (COI) DNA, nuclear ribosomal cluster internal transcribed spacer 1 (ITS1) and ITS2, and Wolbachia-infection status. Our results demonstrated a genetic difference between C. t. altaicus and C. t. sungaris. According to mitochondrial data, the genetic distance between clades of C. t. altaicus and C. t. sungaris was comparable with the species divergence of the genus Callopsylla, which is closely related to Citellophilus. All studied populations of C. t. altaicus were Wolbachia-infected, whereas all studied populations of C. t. sungaris were symbiont-free. Data for ITS1 and ITS2 had much lower phylogenetic signals than mitochondrial data; however, diagnostic substitutions for C. t. altaicus and C. t. sungaris delimitation were also revealed. Analysis of a hardly accessible report on cross experiments allowed us to conclude the partial postzygotic isolation between these subspecies. Taken together, the molecular-genetic, geographical, morphological, and reproductive isolation findings verified that C. t. altaicus and C. t. sungaris subspecies could be considered as different species.}, } @article {pmid35196357, year = {2022}, author = {Ross, PA and Robinson, KL and Yang, Q and Callahan, AG and Schmidt, TL and Axford, JK and Coquilleau, MP and Staunton, KM and Townsend, M and Ritchie, SA and Lau, MJ and Gu, X and Hoffmann, AA}, title = {A decade of stability for wMel Wolbachia in natural Aedes aegypti populations.}, journal = {PLoS pathogens}, volume = {18}, number = {2}, pages = {e1010256}, pmid = {35196357}, issn = {1553-7374}, mesh = {*Aedes ; Animals ; *Arboviruses ; Australia ; *Wolbachia/genetics ; }, abstract = {Mosquitoes carrying Wolbachia endosymbionts are being released in many countries for arbovirus control. The wMel strain of Wolbachia blocks Aedes-borne virus transmission and can spread throughout mosquito populations by inducing cytoplasmic incompatibility. Aedes aegypti mosquitoes carrying wMel were first released into the field in Cairns, Australia, over a decade ago, and with wider releases have resulted in the near elimination of local dengue transmission. The long-term stability of Wolbachia effects is critical for ongoing disease suppression, requiring tracking of phenotypic and genomic changes in Wolbachia infections following releases. We used a combination of field surveys, phenotypic assessments, and Wolbachia genome sequencing to show that wMel has remained stable in its effects for up to a decade in Australian Ae. aegypti populations. Phenotypic comparisons of wMel-infected and uninfected mosquitoes from near-field and long-term laboratory populations suggest limited changes in the effects of wMel on mosquito fitness. Treating mosquitoes with antibiotics used to cure the wMel infection had limited effects on fitness in the next generation, supporting the use of tetracycline for generating uninfected mosquitoes without off-target effects. wMel has a temporally stable within-host density and continues to induce complete cytoplasmic incompatibility. A comparison of wMel genomes from pre-release (2010) and nine years post-release (2020) populations show few genomic differences and little divergence between release locations, consistent with the lack of phenotypic changes. These results indicate that releases of Wolbachia-infected mosquitoes for population replacement are likely to be effective for many years, but ongoing monitoring remains important to track potential evolutionary changes.}, } @article {pmid35192040, year = {2023}, author = {Detcharoen, M and Nilsai, A}, title = {Low Endosymbiont Incidence in Drosophila Species Across Peninsula Thailand.}, journal = {Microbial ecology}, volume = {85}, number = {2}, pages = {730-736}, pmid = {35192040}, issn = {1432-184X}, support = {SCI6404008S//Faculty of Science, Prince of Songkla University/ ; }, mesh = {Animals ; Male ; Drosophila ; Incidence ; Thailand ; Symbiosis ; Bacteroidetes ; *Wolbachia ; *Spiroplasma ; *Rickettsia ; }, abstract = {Arthropods are known to harbor several endosymbionts, such as Cardinium, Rickettsia, Spiroplasma, and Wolbachia. Wolbachia, for example, are the most widespread known endosymbionts in the world, which are found in about half of all arthropod species. To increase their transmission, these endosymbionts must manipulate their hosts in several ways such as cytoplasmic incompatibility and male killing. In tropical regions, endosymbiont diversity has not been studied exhaustively. Here, we checked four endosymbionts, including Cardinium, Rickettsia, Spiroplasma, and Wolbachia, in eleven Drosophila species found in Thai Peninsula. The Wolbachia strain wRi-like was found in all populations of Drosophila ananassae and Drosophila simulans. Furthermore, we found two new strains, wMalA and wMalB, in two populations of Drosophila malerkotliana. Besides Wolbachia, we did not find any of the above endosymbionts in all fly species. This work reveals the hidden diversity of endosymbionts in Drosophila and is the first exhaustive study on Drosophila in the region.}, } @article {pmid35183553, year = {2022}, author = {Konecka, E}, title = {Fifty shades of bacterial endosymbionts and some of them still remain a mystery: Wolbachia and Cardinium in oribatid mites (Acari: Oribatida).}, journal = {Journal of invertebrate pathology}, volume = {189}, number = {}, pages = {107733}, doi = {10.1016/j.jip.2022.107733}, pmid = {35183553}, issn = {1096-0805}, mesh = {Animals ; Bacteria ; Bacteroidetes ; *Mites/microbiology ; Phylogeny ; *Wolbachia ; }, abstract = {Wolbachia is the most abundant intracellular symbiont among terrestrial Arthropoda. This bacterium together with other microorganisms, i.e., Cardinium, gained fame mainly as the causative agent of host sex-ratio distortion. Across the impressive diversity of oribatid mites (Acari: Oribatida), the microbes have been found in both parthenogenetic (Oppiella nova, Ceratozetes thienemanni, Hypochthonius rufulus) as well as sexually-reproducing (Gustavia microcephala, Achipteria coleoptrata, Microzetorchestes emeryi, Damaeus onustus) species. Wolbachia found in Oribatida represents supergroup E and is related to bacterial endosymbionts of springtails (Hexapoda: Collembola). Cardinium identified in O. nova and M. emeryi belongs to phylogenetic group A. In turn, Cardinium from A. coleoptrata constitutes a new separate group E. The occurrence of these bacterial endosymbionts in parthenogenetic and sexual oribatid mites species may suggests a different function other than manipulating host reproduction. Indeed, endosymbionts may have various "shades" of functions in invertebrate hosts, some of which cannot be excluded in the oribatid mites, e.g., enriching a nutrient-limited diet with B vitamins or contributing to host adaptation to colder and harsher climates. Nevertheless, the mystery behind the roles of bacteria in Oribatida still needs required to be unraveled.}, } @article {pmid35175127, year = {2022}, author = {Gharabigloozare, Y and Wähling, A and Bleidorn, C}, title = {Whole-Genome Sequence of the Wolbachia Strain wTcon, an Endosymbiont of the Confused Flour Beetle, Tribolium confusum.}, journal = {Microbiology resource announcements}, volume = {11}, number = {2}, pages = {e0114421}, pmid = {35175127}, issn = {2576-098X}, abstract = {Up to 60% of insects are infected with symbiont intracellular alphaproteobacteria of the genus Wolbachia, which are often able to manipulate their host's reproduction. Here, we report the annotated draft genome sequence of strain wTcon from the confused flour beetle, Tribolium confusum, based on long- and short-read sequence data. The assembled genome is located on 12 contigs with a total size of 1,418,452 bp.}, } @article {pmid35172009, year = {2022}, author = {Rutins, I and Schannauer, S and Orellana, S and Laukhuff, H and Lang, E and Becker, T and McKinney, E and Thomas, K and Tilden, V and Swartz, M and Blair, JE}, title = {Genetic Diversity and Wolbachia (Rickettsiales: Anaplasmataceae) Prevalence Within a Remnant Population of Regal Fritillary, Argynnis idalia (Lepidoptera: Nymphalidae), in South-Central Pennsylvania.}, journal = {Journal of insect science (Online)}, volume = {22}, number = {1}, pages = {}, pmid = {35172009}, issn = {1536-2442}, support = {//Pennsylvania Department of Military and Veterans Affairs/ ; //Bureau of Environmental Management/ ; //Franklin & Marshall College Committee on Grants/ ; }, mesh = {Animals ; *Butterflies/genetics/microbiology ; Female ; Genetic Variation ; Pennsylvania ; Prevalence ; United States ; *Wolbachia/genetics ; }, abstract = {Eastern populations of the North American regal fritillary, Argynnis idalia Drury (1773), have been largely extirpated over the past half century. Here we report on the last remaining population of eastern regal fritillaries, located within a military installation in south-central Pennsylvania. Samples were obtained from field specimens during two years of annual monitoring, and from females collected for captive rearing over a five year period. Nuclear microsatellite and mitochondrial sequence data do not suggest subdivision within this population, but excess nuclear homozygosity indicates negative impacts on genetic diversity likely due to small population size and potential inbreeding effects. Molecular assays did not detect Wolbachia endosymbionts in field specimens of regal fritillary, but sympatric Argynnis sister species showed high prevalence of Wolbachia infected individuals. Our results inform ongoing conservation and reintroduction projects, designed to protect the last remaining regal fritillary population from extirpation in the eastern United States.}, } @article {pmid35171977, year = {2022}, author = {Gagalova, KK and Whitehill, JGA and Culibrk, L and Lin, D and Lévesque-Tremblay, V and Keeling, CI and Coombe, L and Yuen, MMS and Birol, I and Bohlmann, J and Jones, SJM}, title = {The genome of the forest insect pest Pissodes strobi reveals genome expansion and evidence of a Wolbachia endosymbiont.}, journal = {G3 (Bethesda, Md.)}, volume = {12}, number = {4}, pages = {}, pmid = {35171977}, issn = {2160-1836}, mesh = {Animals ; Forests ; Insecta ; *Picea/genetics ; *Weevils/genetics ; *Wolbachia/genetics ; }, abstract = {The highly diverse insect family of true weevils, Curculionidae, includes many agricultural and forest pests. Pissodes strobi, commonly known as the spruce weevil or white pine weevil, is a major pest of spruce and pine forests in North America. Pissodes strobi larvae feed on the apical shoots of young trees, causing stunted growth and can destroy regenerating spruce or pine forests. Here, we describe the nuclear and mitochondrial Pissodes strobi genomes and their annotations, as well as the genome of an apparent Wolbachia endosymbiont. We report a substantial expansion of the weevil nuclear genome, relative to other Curculionidae species, possibly driven by an abundance of class II DNA transposons. The endosymbiont observed belongs to a group (supergroup A) of Wolbachia species that generally form parasitic relationships with their arthropod host.}, } @article {pmid35166193, year = {2022}, author = {Yan, R and Sun, Q}, title = {Uniqueness and stability of periodic solutions for an interactive wild and Wolbachia-infected male mosquito model.}, journal = {Journal of biological dynamics}, volume = {16}, number = {1}, pages = {254-276}, doi = {10.1080/17513758.2022.2037760}, pmid = {35166193}, issn = {1751-3766}, mesh = {Animals ; *Culicidae ; Male ; Models, Biological ; Mosquito Vectors ; *Wolbachia ; }, abstract = {We investigate a mosquito population suppression model, which includes the release of Wolbachia-infected males causing incomplete cytoplasmic incompatibility (CI). The model consists of two sub-equations by considering the density-dependent birth rate of wild mosquitoes. By assuming the release waiting period T is larger than the sexual lifespan T¯ of Wolbachia-infected males, we derive four thresholds: the CI intensity threshold sh∗, the release amount thresholds g∗ and c∗, and the waiting period threshold T∗. From a biological view, we assume sh>sh∗ throughout the paper. When g∗T∗, which is globally asymptotically stable. Our theoretical results are confirmed by numerical simulations.}, } @article {pmid35162074, year = {2022}, author = {Skinner, KM and Underwood, J and Ghosh, A and Oliva Chavez, AS and Brelsfoard, CL}, title = {Wolbachia Impacts Anaplasma Infection in Ixodes scapularis Tick Cells.}, journal = {International journal of environmental research and public health}, volume = {19}, number = {3}, pages = {}, pmid = {35162074}, issn = {1660-4601}, mesh = {*Anaplasma phagocytophilum ; *Anaplasmosis ; Animals ; Host-Pathogen Interactions ; *Ixodes/microbiology ; *Wolbachia ; }, abstract = {The specific interactions of members of tick bacterial microbiota and their effects on pathogen transmission remains relatively unexplored. Here, we introduced a novel Wolbachia infection type into Ixodes scapularis tick cells and examined the antipathogenic effects on the intracellular pathogen Anaplasma phagocytophilum. An increase in A. phagocytophilum replication was observed in Wolbachia-infected tick cells. However, Wolbachia infection densities decreased when cells were serially passaged and ultimately the infection was lost. Host-cell immune response was also examined as an additional factor that could have affected A. phagocytophilum replication in Wolbachia-infected cells. In early passages post-Wolbachia infection, a decreased immune response was observed, but in later passages of cells with low Wolbachia densities, there was no change in the immune response. The results are discussed in relation to the importance of studying the interactions of the tick microbiota, the host cell, and the pathogen and the development of novel tick and tick-borne disease-control approaches.}, } @article {pmid35140881, year = {2022}, author = {Schrieke, H and Maignien, L and Constancias, F and Trigodet, F and Chakloute, S and Rakotoarivony, I and Marie, A and L'Ambert, G and Makoundou, P and Pages, N and Murat Eren, A and Weill, M and Sicard, M and Reveillaud, J}, title = {The mosquito microbiome includes habitat-specific but rare symbionts.}, journal = {Computational and structural biotechnology journal}, volume = {20}, number = {}, pages = {410-420}, pmid = {35140881}, issn = {2001-0370}, abstract = {Microbial communities are known to influence mosquito lifestyles by modifying essential metabolic and behavioral processes that affect reproduction, development, immunity, digestion, egg survival, and the ability to transmit pathogens. Many studies have used 16S rRNA gene amplicons to characterize mosquito microbiota and investigate factors that influence host-microbiota dynamics. However, a relatively low taxonomic resolution due to clustering methods based on arbitrary threshold and the overall dominance of Wolbachia or Asaia symbionts obscured the investigation of rare members of mosquito microbiota in previous studies. Here, we used high resolution Shannon entropy-based oligotyping approach to analyze the microbiota of Culex pipiens, Culex quinquefasciatus and Aedes individuals from continental Southern France and overseas Guadeloupe as well as from laboratories with or without antibiotics treatment. Our experimental design that resulted in a series of mosquito samples with a gradient of Wolbachia density and relative abundance along with high-resolution analyses of amplicon sequences enabled the recovery of a robust signal from typically less accessible bacterial taxa. Our data confirm species-specific mosquito-bacteria associations with geography as a primary factor that influences bacterial community structure. But interestingly, they also reveal co-occurring symbiotic bacterial variants within single individuals for both Elizabethkingia and Erwinia genera, distinct and specific Asaia and Chryseobacterium in continental and overseas territories, and a putative rare Wolbachia variant. Overall, our study reveals the presence of previously overlooked microdiversity and multiple closely related symbiotic strains within mosquito individuals with a remarkable habitat-specificity.}, } @article {pmid35135613, year = {2022}, author = {Zhang, XL and Deng, YP and Yang, T and Li, LY and Cheng, TY and Liu, GH and Duan, DY}, title = {Metagenomics of the midgut microbiome of Rhipicephalus microplus from China.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {48}, pmid = {35135613}, issn = {1756-3305}, support = {31902294//the National Science Foundation of China/ ; 2018RS3085//the Planned Programme of Hunan Province Science and Technology Innovation/ ; KH2002001//the Training Programme for Excellent Young Innovators of Changsha/ ; 2020JJ5230//the Natural Science Foundation of Hunan Province, China/ ; 19A235//the Research Foundation of Education Bureau of Hunan Province, China/ ; }, mesh = {*Anaplasmosis ; Animals ; Cattle ; *Cattle Diseases ; Female ; Metagenomics ; *Microbiota/genetics ; *Rhipicephalus/genetics ; *Tick Infestations/veterinary ; *Tick-Borne Diseases ; }, abstract = {BACKGROUND: Ticks, which are ectoparasites of animals, may carry multiple pathogens. The cattle tick Rhipicephalus microplus is an important bovine parasite in China. However, the midgut microbiome of R. microplus from China has not been characterized via metagenomic methods.

METHODS: Rhipicephalus microplus were collected from cattle in the city of Changsha in Hunan province, China. The DNA of the midgut contents was extracted from fully engorged adult female R. microplus. A DNA library was constructed and sequenced using an Illumina HiSeq sequencing platform. SOAPdenovo software was used to assemble and analyze the clean data. The latent class analysis algorithm applied to system classification by MEGAN software was used to annotate the information on the species' sequences. DIAMOND software was used to compare unigenes with the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and functional annotation was carried out based on the results of the comparison.

RESULTS: The dominant phyla in the five samples were Firmicutes, Proteobacteria, and Actinobacteria. Streptococcus, Mycobacterium, Anaplasma, Enterococcus, Shigella, Lactobacillus, Brachyspira, Pseudomonas, Enterobacter, Bacillus, and Lactococcus were the dominant genera in the five samples. The endosymbiotic bacterium Wolbachia was also detected in all of the samples. Mycobacterium malmesburyense, Streptococcus pneumoniae, Anaplasma phagocytophilum, Enterococcus faecium, Shigella sonnei, Enterococcus faecalis, Lactobacillus casei, Brachyspira hampsonii, Pseudomonas syringae, Enterobacter cloacae, and Lactococcus garvieae were the dominant species in the five samples. In addition to these bacterial species, we also detected some eukaryotes, such as Rhizophagus irregularis, Enterospora canceri, Smittium culicis, Zancudomyces culisetae, Trachipleistophora hominis, and viruses such as orf virus, human endogenous retrovirus type W, enzootic nasal tumor virus of goats, bovine retrovirus CH15, and galidia endogenous retrovirus in all of the samples at the species level. The results of the annotated KEGG pathway predictions for the gene functions of the midgut microflora of R. microplus indicated genes involved in lipid and amino acid metabolism, infectious diseases (e.g., Streptococcus pneumonia infection, human granulocytic anaplasmosis, Shigella sonnei infection, Salmonella enterica infection, and pathogenic Escherichia coli infection), and cancer.

CONCLUSIONS: Our study revealed that the midgut microbiome of R. microplus is not only composed of a large number of bacteria, but that a portion also comprises eukaryotes and viruses. The data presented here enhance our understanding of this tick's midgut microbiome and provide fundamental information for the control of ticks and tick-borne diseases.}, } @article {pmid35134330, year = {2022}, author = {Horard, B and Terretaz, K and Gosselin-Grenet, AS and Sobry, H and Sicard, M and Landmann, F and Loppin, B}, title = {Paternal transmission of the Wolbachia CidB toxin underlies cytoplasmic incompatibility.}, journal = {Current biology : CB}, volume = {32}, number = {6}, pages = {1319-1331.e5}, doi = {10.1016/j.cub.2022.01.052}, pmid = {35134330}, issn = {1879-0445}, mesh = {Animals ; *Culex/genetics ; Cytoplasm ; Cytosol ; Drosophila ; Male ; *Wolbachia/genetics ; }, abstract = {Wolbachia are widespread endosymbiotic bacteria that manipulate the reproduction of arthropods through a diversity of cellular mechanisms. In cytoplasmic incompatibility (CI), a sterility syndrome originally discovered in the mosquito Culex pipiens, uninfected eggs fertilized by sperm from infected males are selectively killed during embryo development following the abortive segregation of paternal chromosomes in the zygote. Despite the recent discovery of Wolbachia CI factor (cif) genes, the mechanism by which they control the fate of paternal chromosomes at fertilization remains unknown. Here, we have analyzed the cytological distribution and cellular impact of CidA and CidB, a pair of Cif proteins from the Culex-infecting Wolbachia strain wPip. We show that expression of CidB in Drosophila S2R+ cells induces apoptosis unless CidA is co-expressed and associated with its partner. In transgenic Drosophila testes, both effectors colocalize in germ cells until the histone-to-protamine transition in which only CidB is retained in maturing spermatid nuclei. We further show that CidB is similarly targeted to maturing sperm of naturally infected Culex mosquitoes. At fertilization, CidB associates with paternal DNA regions exhibiting DNA replication stress, as a likely cause of incomplete replication of paternal chromosomes at the onset of the first mitosis. Importantly, we demonstrate that inactivation of the deubiquitylase activity of CidB does not abolish its cell toxicity or its ability to induce CI in Drosophila. Our study thus demonstrates that CI functions as a transgenerational toxin-antidote system and suggests that CidB acts by poisoning paternal DNA replication in incompatible crosses.}, } @article {pmid35134226, year = {2022}, author = {Colgan, TJ and Arce, AN and Gill, RJ and Ramos Rodrigues, A and Kanteh, A and Duncan, EJ and Li, L and Chittka, L and Wurm, Y}, title = {Genomic Signatures of Recent Adaptation in a Wild Bumblebee.}, journal = {Molecular biology and evolution}, volume = {39}, number = {2}, pages = {}, pmid = {35134226}, issn = {1537-1719}, support = {BB/K004204/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/T015683/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Bees/genetics ; *Ecosystem ; *Genomics ; }, abstract = {Environmental changes threaten insect pollinators, creating risks for agriculture and ecosystem stability. Despite their importance, we know little about how wild insects respond to environmental pressures. To understand the genomic bases of adaptation in an ecologically important pollinator, we analyzed genomes of Bombus terrestris bumblebees collected across Great Britain. We reveal extensive genetic diversity within this population, and strong signatures of recent adaptation throughout the genome affecting key processes including neurobiology and wing development. We also discover unusual features of the genome, including a region containing 53 genes that lacks genetic diversity in many bee species, and a horizontal gene transfer from a Wolbachia bacteria. Overall, the genetic diversity we observe and how it is distributed throughout the genome and the population should support the resilience of this important pollinator species to ongoing and future selective pressures. Applying our approach to more species should help understand how they can differ in their adaptive potential, and to develop conservation strategies for those most at risk.}, } @article {pmid35132164, year = {2022}, author = {Xu, W and Wang, Y and Wang, YH and Zhang, YN and Wang, JF}, title = {Diversity and dynamics of bacteria at the Chrysomya megacephala pupal stage revealed by third-generation sequencing.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {2006}, pmid = {35132164}, issn = {2045-2322}, mesh = {Animals ; Bacteroidetes/genetics/isolation & purification/physiology ; Calliphoridae/*microbiology ; Erysipelothrix/genetics/isolation & purification ; Firmicutes/genetics/isolation & purification/physiology ; Forensic Entomology ; Gammaproteobacteria/genetics/isolation & purification/physiology ; *Microbiota/genetics/physiology ; Proteobacteria/genetics/isolation & purification/physiology ; Pupa/*microbiology ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, RNA/*methods ; Symbiosis ; Wolbachia/genetics/isolation & purification/physiology ; }, abstract = {Characterization of the microbial community is essential for understanding the symbiotic relationships between microbes and host insects. Chrysomya megacephala is a vital resource, a forensic insect, a pollinator, and a vector for enteric bacteria, protozoa, helminths, and viruses. However, research on its microbial community is incomprehensive, particularly at the pupal stage, which comprises approximately half of the entire larval development stage and is important entomological evidence in forensic medicine. For the first time, this study investigated the bacterial communities of C. megacephala pupae at different ages using third-generation sequencing technology. The results showed that C. megacephala has a diverse and dynamic bacterial community. Cluster analysis at ≥ 97% similarity produced 154 operational taxonomic units (OTUs) that belonged to 10 different phyla and were distributed into 15 classes, 28 orders, 50 families, 88 genera, and 130 species. Overall, the number of bacterial OTUs increased with the development of pupae, and the relative abundance of Wolbachia in the Day5 group was significantly lower than that in the other groups. Within the pupal stage, Proteobacteria, Firmicutes, and Bacteroidetes were the dominant phyla of bacteria. At the genus level, Wolbachia and Ignatzschineria coexisted, a rarely known feature. In addition, we found Erysipelothrix rhusiopathiae, the etiological agent of swine erysipelas, which is rarely identified in insects. This study enriches the understanding of the microbial community of C. megacephala and provides a reference for better utilization and control of C. megacephala.}, } @article {pmid35127567, year = {2021}, author = {Mao, W and Zeng, Q and She, L and Yuan, H and Luo, Y and Wang, R and She, Y and Wang, W and Wang, C and Pan, X}, title = {Wolbachia Utilizes lncRNAs to Activate the Anti-Dengue Toll Pathway and Balance Reactive Oxygen Species Stress in Aedes aegypti Through a Competitive Endogenous RNA Network.}, journal = {Frontiers in cellular and infection microbiology}, volume = {11}, number = {}, pages = {823403}, pmid = {35127567}, issn = {2235-2988}, mesh = {*Aedes/genetics ; Animals ; Gene Regulatory Networks ; *MicroRNAs/genetics ; *RNA, Long Noncoding/genetics/metabolism ; RNA, Messenger/genetics ; Reactive Oxygen Species ; *Wolbachia/genetics ; }, abstract = {Long non-coding RNAs (lncRNA), a class of RNA molecules without protein coding potential, are more than 200 nucleotides in length and widely present in a variety of species. Although increasing progress in regard to the determination of lncRNA function has been made in vertebrates, Aedes aegypti lncRNAs were only identified recently and the functions of few lncRNAs have been annotated so far. Herein, the genome-wide alteration of the lncRNA expression profile trigged by Wolbachia wAlbB infection was investigated by comparing A. aegypti Aag2 cells and W-Aag2 cells infected with Wolbachia wAlbB. Based on lncRNA sequencing, 3035 differentially expressed lncRNAs (DE lncRNAs) in total were identified upon Wolbachia infection, which were further validated by quantitative PCR. The constructed co-expression network of DE lncRNAs and mRNAs revealed that Wolbachia-induced DE lncRNAs were highly enriched in the oxidative phosphorylation pathway via trans-activity, according to the KEGG pathway enrichment analyses. In addition, the established competitive endogenous RNA (ceRNA) network identifies the DE lncRNAs enriched in cellular oxidant detoxification based on GO enrichment analysis. Furthermore, silencing of aae-lnc-7598, the significantly up-regulated lncRNA with the highest fold change induced by Wolbachia, caused a significant reduction of antioxidant catalase 1B (CAT1B) gene expression as well as the enhancement of mitochondrial reactive oxygen species (ROS) production in living cells. These findings indicate that Wolbachia manipulates lncRNA to balance intracellular ROS stress and ensure its endosymbiosis in host A. aegypti. Notably, the function assay demonstrated that aae-lnc-0165 suppressed by Wolbachia could induce expression of the REL1 gene, the key regulator of downstream Toll pathway, through the sequence-specific binding of aae-miR-980-5p, which contributes to the activation of Toll pathway. Moreover, the depletion of aae-lnc-0165 caused the suppression of mitochondrial ROS levels in living cells. Our data reveal that Wolbachia activates the anti-dengue Toll pathway through a lncRNA-ceRNA pattern. Taken together, our finding suggested that Wolbachia utilizes lncRNAs to activate host anti-dengue Toll pathway via a ceRNA network. Moreover, Wolbachia employs lncRNAs to ensure ROS homeostasis for ROS-based anti-dengue defense through either trans-regulation or the ceRNA network. This study identifies novel potential molecular biomarkers for prevention and control of epidemic dengue.}, } @article {pmid35127140, year = {2022}, author = {Turelli, M and Barton, NH}, title = {Why did the Wolbachia transinfection cross the road? drift, deterministic dynamics, and disease control.}, journal = {Evolution letters}, volume = {6}, number = {1}, pages = {92-105}, pmid = {35127140}, issn = {2056-3744}, abstract = {Maternally inherited Wolbachia transinfections are being introduced into natural mosquito populations to reduce the transmission of dengue, Zika, and other arboviruses. Wolbachia-induced cytoplasmic incompatibility provides a frequency-dependent reproductive advantage to infected females that can spread transinfections within and among populations. However, because transinfections generally reduce host fitness, they tend to spread within populations only after their frequency exceeds a critical threshold. This produces bistability with stable equilibrium frequencies at both 0 and 1, analogous to the bistability produced by underdominance between alleles or karyotypes and by population dynamics under Allee effects. Here, we analyze how stochastic frequency variation produced by finite population size can facilitate the local spread of variants with bistable dynamics into areas where invasion is unexpected from deterministic models. Our exemplar is the establishment of wMel Wolbachia in the Aedes aegypti population of Pyramid Estates (PE), a small community in far north Queensland, Australia. In 2011, wMel was stably introduced into Gordonvale, separated from PE by barriers to A. aegypti dispersal. After nearly 6 years during which wMel was observed only at low frequencies in PE, corresponding to an apparent equilibrium between immigration and selection, wMel rose to fixation by 2018. Using analytic approximations and statistical analyses, we demonstrate that the observed fixation of wMel at PE is consistent with both stochastic transition past an unstable threshold frequency and deterministic transformation produced by steady immigration at a rate just above the threshold required for deterministic invasion. The indeterminacy results from a delicate balance of parameters needed to produce the delayed transition observed. Our analyses suggest that once Wolbachia transinfections are established locally through systematic introductions, stochastic "threshold crossing" is likely to only minimally enhance spatial spread, providing a local ratchet that slightly-but systematically-aids area-wide transformation of disease-vector populations in heterogeneous landscapes.}, } @article {pmid35126329, year = {2021}, author = {Zheng, Z and Zhao, M and Zhang, Z and Hu, X and Xu, Y and Wei, C and He, H}, title = {Lactic Acid Bacteria Are Prevalent in the Infrabuccal Pockets and Crops of Ants That Prefer Aphid Honeydew.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {785016}, pmid = {35126329}, issn = {1664-302X}, abstract = {Ants are evolutionarily successful species and occupy diverse trophic and habitat niches on the earth. To fulfill dietary requirements, ants have established commensalism with both sap-feeding insects and bacteria. In this study, we used high-throughput sequencing of the bacterial 16S rRNA gene to characterize the bacterial composition and structure of the digestive tracts in three species of Formica ants and Lasius niger (Linnaeus)-species that predominantly feed on honeydew secreted by aphids. We found that bacterial communities displayed species- and colony-level signatures, and that bacterial communities in the infrabuccal pockets and crops were different from those in the midguts and hindguts. Lactobacillus and Wolbachia were dominant in the infrabuccal pockets and crops of workers, whereas Wolbachia was dominant in the midguts, hindguts and brood (larvae, pupae and cocoons). To learn more about the dominant Lactobacillus in ants, we assessed its prevalence in a wide range of aphid-tending ants using diagnostic PCR. We found that Lactobacillus was more prevalent in Formicinae than in Myrmicinae species. We also isolated four strains of lactic acid bacteria (Lactobacillus sanfranciscensis, Lactobacillus lindneri, Weissella cibaria and Fructobacillus sp.) from the infrabuccal pockets and crops of aphid-tending ants using a culture-dependent method. Two predominant lactic acid bacterial isolates, Lactobacillus sanfranciscensis (La2) and Weissella cibaria (La3), exhibited abilities in catabolizing sugars (sucrose, trehalose, melezitose and raffinose) known to be constituents of hemipteran honeydew. These findings contribute to further understanding the association between ants, aphids and bacteria, and provide additional information on the function of lactic acid bacteria in ants.}, } @article {pmid35124699, year = {2022}, author = {Doremus, MR and Stouthamer, CM and Kelly, SE and Schmitz-Esser, S and Hunter, MS}, title = {Quality over quantity: unraveling the contributions to cytoplasmic incompatibility caused by two coinfecting Cardinium symbionts.}, journal = {Heredity}, volume = {128}, number = {3}, pages = {187-195}, pmid = {35124699}, issn = {1365-2540}, mesh = {Animals ; Bacteroidetes/genetics ; Cytoplasm/microbiology ; Female ; Male ; Reproduction ; Symbiosis ; *Wasps/genetics/microbiology ; *Wolbachia ; }, abstract = {Cytoplasmic incompatibility (CI) is a common form of reproductive sabotage caused by maternally inherited bacterial symbionts of arthropods. CI is a two-step manipulation: first, the symbiont modifies sperm in male hosts which results in the death of fertilized, uninfected embryos. Second, when females are infected with a compatible strain, the symbiont reverses sperm modification in the fertilized egg, allowing offspring of infected females to survive and spread the symbiont to high frequencies in a population. Although CI plays a role in arthropod evolution, the mechanism of CI is unknown for many symbionts. Cardinium hertigii is a common CI-inducing symbiont of arthropods, including parasitoid wasps like Encarsia partenopea. This wasp harbors two Cardinium strains, cEina2 and cEina3, and exhibits strong CI. The strains infect wasps at different densities, with the cEina3 present at a lower density than cEina2, and it was previously not known which strain caused CI. By differentially curing wasps of cEina3, we found that this low-density symbiont is responsible for CI and modifies males during their pupal stage. cEina2 does not modify host reproduction and may spread by 'hitchhiking' with cEina3 CI or by conferring an unknown benefit. The cEina3 strain also shows a unique localization pattern in male reproductive tissues. Instead of infecting sperm like other CI-inducing symbionts, cEina3 cells are found in somatic cells at the testis base and around the seminal vesicle. This may allow the low-density cEina3 to efficiently modify host males and suggests that cEina3 uses a different modification strategy than sperm-infecting CI symbionts.}, } @article {pmid35124068, year = {2022}, author = {Alonso, AC and Stein, M and Matías Hisgen, C and Micieli, MV}, title = {Abiotic factors affecting the prevalence of Wolbachia (Rickettsiaceae) in immature Aedes albopictus (Skuse) (Culicidae).}, journal = {Journal of invertebrate pathology}, volume = {189}, number = {}, pages = {107730}, doi = {10.1016/j.jip.2022.107730}, pmid = {35124068}, issn = {1096-0805}, mesh = {*Aedes/microbiology ; Animals ; Prevalence ; *Rickettsiaceae ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia is a genus of gram-negative endosymbiotic bacterium of maternal transmission, located mainly in the gonads of arthropods, including mosquitoes such as Aedes albopictus. The current distribution of Ae. albopictus in Argentina is restricted to the subtropical northeastern region of the country. Here, we studied the seasonal prevalence of Wolbachia detected in Ae. albopictus larvae and the relationship between the abiotic factors of the larval microhabitat and the infection status, in Eldorado city, Misiones province, subtropical region. The prevalence of Wolbachia infection found was 76.89% (n = 312). From the total samples examined, 52.80% (n = 214) showed double infection with the wAlbA/wAlbB strains, 23.84% (n = 97) infection only with wAlbB, and 0.25% (n = 1) only with wAlbA. The prevalence of double infection did not present statistically significant differences between the sites studied. For single infection, the lowest prevalence value of the wAlbB strain (13.33%) was found in the natural park, whereas the highest was found in the family dwellings and cemeteries. Tire repair shops showed an intermediate value. The wAlbA single infection was identified once. Our results also showed an association between temperature and slightly turbid waters with exposure to the sun in the larval habitats and the probability of infection by Wolbachia.}, } @article {pmid35116014, year = {2021}, author = {Gendrin, M and Favia, G and Herren, JK}, title = {Editorial: Microbiota: A Consequential Third Wheel in the Mosquito-Pathogen Relationship.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {811880}, doi = {10.3389/fmicb.2021.811880}, pmid = {35116014}, issn = {1664-302X}, } @article {pmid35115867, year = {2022}, author = {Raupach, MJ and Rulik, B and Spelda, J}, title = {Surprisingly high genetic divergence of the mitochondrial DNA barcode fragment (COI) within Central European woodlice species (Crustacea, Isopoda, Oniscidea).}, journal = {ZooKeys}, volume = {1082}, number = {}, pages = {103-125}, pmid = {35115867}, issn = {1313-2989}, abstract = {DNA barcoding has become the most popular approach for species identification in recent years. As part of the German Barcode of Life project, the first DNA barcode library for terrestrial and freshwater isopods from Germany is presented. The analyzed barcode library included 38 terrestrial (78% of the documented species of Germany) and five freshwater (63%) species. A total of 513 new barcodes was generated and 518 DNA barcodes were analyzed. This analysis revealed surprisingly high intraspecific genetic distances for numerous species, with a maximum of 29.4% for Platyarthrushoffmannseggii Brandt, 1833. The number of BINs per species ranged from one (32 species, 68%) to a maximum of six for Trachelipusrathkii (Brandt, 1833). In spite of such high intraspecific variability, interspecific distances with values between 12.6% and 29.8% allowed a valid species assignment of all analyzed isopods. The observed high intraspecific distances presumably result from phylogeographic events, Wolbachia infections, atypical mitochondrial DNAs, heteroplasmy, or various combinations of these factors. Our study represents the first step in generating an extensive reference library of DNA barcodes for terrestrial and freshwater isopods for future molecular biodiversity assessment studies.}, } @article {pmid35115648, year = {2022}, author = {Katlav, A and Nguyen, DT and Morrow, JL and Spooner-Hart, RN and Riegler, M}, title = {Endosymbionts moderate constrained sex allocation in a haplodiploid thrips species in a temperature-sensitive way.}, journal = {Heredity}, volume = {128}, number = {3}, pages = {169-177}, pmid = {35115648}, issn = {1365-2540}, mesh = {Animals ; Bacteroidetes ; Female ; Male ; Sex Ratio ; Symbiosis/genetics ; Temperature ; *Thysanoptera/genetics/microbiology ; *Wolbachia/genetics ; }, abstract = {Maternally inherited bacterial endosymbionts that affect host fitness are common in nature. Some endosymbionts colonise host populations by reproductive manipulations (such as cytoplasmic incompatibility; CI) that increase the reproductive fitness of infected over uninfected females. Theory predicts that CI-inducing endosymbionts in haplodiploid hosts may also influence sex allocation, including in compatible crosses, however, empirical evidence for this is scarce. We examined the role of two common CI-inducing endosymbionts, Cardinium and Wolbachia, in the sex allocation of Pezothrips kellyanus, a haplodiploid thrips species with a split sex ratio. In this species, irrespective of infection status, some mated females are constrained to produce extremely male-biased broods, whereas other females produce extremely female-biased broods. We analysed brood sex ratio of females mated with males of the same infection status at two temperatures. We found that at 20 °C the frequency of constrained sex allocation in coinfected pairs was reduced by 27% when compared to uninfected pairs. However, at 25 °C the constrained sex allocation frequency increased and became similar between coinfected and uninfected pairs, resulting in more male-biased population sex ratios at the higher temperature. This temperature-dependent pattern occurred without changes in endosymbiont densities and compatibility. Our findings indicate that endosymbionts affect sex ratios of haplodiploid hosts beyond the commonly recognised reproductive manipulations by causing female-biased sex allocation in a temperature-dependent fashion. This may contribute to a higher transmission efficiency of CI-inducing endosymbionts and is consistent with previous models that predict that CI by itself is less efficient in driving endosymbiont invasions in haplodiploid hosts.}, } @article {pmid35112745, year = {2022}, author = {Sigle, LT and Jones, M and Novelo, M and Ford, SA and Urakova, N and Lymperopoulos, K and Sayre, RT and Xi, Z and Rasgon, JL and McGraw, EA}, title = {Assessing Aedes aegypti candidate genes during viral infection and Wolbachia-mediated pathogen blocking.}, journal = {Insect molecular biology}, volume = {31}, number = {3}, pages = {356-368}, pmid = {35112745}, issn = {1365-2583}, support = {R01 AI143758/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes/genetics ; Animals ; *Chikungunya virus ; *Dengue Virus/genetics ; Mosquito Vectors/genetics ; *Virus Diseases ; *Wolbachia/physiology ; }, abstract = {One approach to control dengue virus transmission is the symbiont Wolbachia, which limits viral infection in mosquitoes. Despite plans for its widespread use in Aedes aegypti, Wolbachia's mode of action remains poorly understood. Many studies suggest that the mechanism is likely multifaceted, involving aspects of immunity, cellular stress and nutritional competition. A previous study from our group used artificial selection to identify a new mosquito candidate gene related to viral blocking; alpha-mannosidase-2a (alpha-Mann-2a) with a predicted role in protein glycosylation. Protein glycosylation pathways tend to be involved in complex host-viral interactions; however, the function of alpha-mannosidases has not been described in mosquito-virus interactions. We examined alpha-Mann-2a expression in response to virus and Wolbachia infections and whether reduced gene expression, caused by RNA interference, affected viral loads. We show that dengue virus (DENV) infection affects the expression of alpha-Mann-2a in a tissue- and time-dependent manner, whereas Wolbachia infection had no effect. In the midgut, DENV prevalence increased following knockdown of alpha-Mann-2a expression in Wolbachia-free mosquitoes, suggesting that alpha-Mann-2a interferes with infection. Expression knockdown had the same effect on the togavirus chikungunya virus, indicating that alpha-Mann-2a may have broad antivirus effects in the midgut. Interestingly, we were unable to knockdown the expression in Wolbachia-infected mosquitoes. We also provide evidence that alpha-Mann-2a may affect the transcriptional level of another gene predicted to be involved in viral blocking and cell adhesion; cadherin87a. These data support the hypothesis that glycosylation and adhesion pathways may broadly be involved in viral infection in Ae. aegypti.}, } @article {pmid35092614, year = {2022}, author = {Rodrigues, LR and Zélé, F and Santos, I and Magalhães, S}, title = {No evidence for the evolution of mating behavior in spider mites due to Wolbachia-induced cytoplasmic incompatibility.}, journal = {Evolution; international journal of organic evolution}, volume = {76}, number = {3}, pages = {623-635}, doi = {10.1111/evo.14429}, pmid = {35092614}, issn = {1558-5646}, support = {COMPCON GA 725419//H2020 European Research Council/ ; SFRH/BD/87628/2012//Fundação para a Ciência e a Tecnologia/ ; EXPL/BIA-EVL/0131/2021//Fundação para a Ciência e a Tecnologia/ ; FCT-ANR//BIA- EVF/0013/2012//FCT-ANR collaboration/ ; }, mesh = {Animals ; *Arthropods ; Cytoplasm ; Female ; Male ; Reproduction ; *Tetranychidae/genetics ; *Wolbachia/genetics ; }, abstract = {Arthropods are often infected with Wolbachia inducing cytoplasmic incompatibility (CI), whereby crosses between uninfected females and infected males yield unviable fertilized offspring. Although uninfected females benefit from avoiding mating with Wolbachia-infected males, this behavior is not always present in host populations and its evolution may hinge upon various factors. Here, we used spider mites to test whether CI could select for mate preference in uninfected females in absence of kin recognition. We found that uninfected females from several field-derived populations showed no preference for infected or uninfected males, nor evolved a preference after being exposed to CI for 12-15 generations by maintaining uninfected females with both infected and uninfected males (i.e., stable "infection polymorphism"). This suggests that Wolbachia-mediated mate choice evolution may require very specific conditions in spider mites. However, after experimental evolution, the copulation duration of Wolbachia-infected control males was significantly higher than that of uninfected control males, but not than that of uninfected males from the "infection polymorphism" regime. This result illustrates how gene flow may oppose Wolbachia-driven divergence between infected and uninfected hosts in natural populations.}, } @article {pmid35073749, year = {2022}, author = {Sun, G and Zhang, M and Chen, H and Hochstrasser, M}, title = {The CinB Nuclease from wNo Wolbachia Is Sufficient for Induction of Cytoplasmic Incompatibility in Drosophila.}, journal = {mBio}, volume = {13}, number = {1}, pages = {e0317721}, pmid = {35073749}, issn = {2150-7511}, support = {R35 GM136325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Male ; Female ; Humans ; Drosophila/microbiology ; *Wolbachia/genetics ; Saccharomyces cerevisiae ; Animals, Genetically Modified ; Cytoplasm/microbiology ; *Infertility, Male ; }, abstract = {Wolbachia is an obligate intracellular bacterium that can alter reproduction of its arthropod hosts, often through a mechanism called cytoplasmic incompatibility (CI). In CI, uninfected females fertilized by infected males yield few offspring, but if both are similarly infected, normal embryo viability results (called "rescue"). CI factors (Cifs) responsible for CI are pairs of proteins encoded by linked genes. The downstream gene in each pair encodes either a deubiquitylase (CidB) or a nuclease (CinB). The upstream gene products, CidA and CinA, bind their cognate enzymes with high specificity. Expression of CidB or CinB in yeast inhibits growth, but growth is rescued by expression of the cognate CifA protein. By contrast, transgenic Drosophila male germ line expression of both cifA and cifB was reported to be necessary to induce CI-like embryonic arrest; cifA expression alone in females is sufficient for rescue. This pattern, seen with genes from several Wolbachia strains, has been called the "2-by-1" model. Here, we show that male germ line expression of the cinB gene alone, from a distinct clade of cif genes from wNo Wolbachia, is sufficient to induce nearly complete loss of embryo viability. This male sterility is fully rescued by cognate cinA[w][No] expression in the female germ line. The proteins behave similarly in yeast. CinB[w][No] toxicity depends on its nuclease active site. These results demonstrate that highly divergent CinB nucleases can induce CI, that rescue by cognate CifA factors is a general feature of Wolbachia CI systems, and that CifA is not strictly required in males for CI induction. IMPORTANCE Wolbachia bacteria live within the cells of many insects. Like mitochondria, they are only inherited from females. Wolbachia often increases the number of infected females to promote spread of infection using a type of male sterility called cytoplasmic incompatibility (CI): when uninfected females mate with infected males, most embryos die; if both are similarly infected, embryos develop normally, giving infected females an advantage in producing offspring. CI is being used against disease-carrying mosquitoes and agricultural pests. Wolbachia proteins called CifA and CifB, which bind one another, cause CI, but how they work has been unclear. Here, we show that a CifB protein singly produced in fruit fly males causes sterility in crosses to normal females, but this is rescued if the females produce the CifA partner. These findings clarify a broad range of observations on CI and will allow more rational approaches to using it for insect control.}, } @article {pmid35069682, year = {2021}, author = {Rašić, G and Lobo, NF and Jeffrey Gutiérrez, EH and Sánchez C, HM and Marshall, JM}, title = {Monitoring Needs for Gene Drive Mosquito Projects: Lessons From Vector Control Field Trials and Invasive Species.}, journal = {Frontiers in genetics}, volume = {12}, number = {}, pages = {780327}, pmid = {35069682}, issn = {1664-8021}, abstract = {As gene drive mosquito projects advance from contained laboratory testing to semi-field testing and small-scale field trials, there is a need to assess monitoring requirements to: i) assist with the effective introduction of the gene drive system at field sites, and ii) detect unintended spread of gene drive mosquitoes beyond trial sites, or resistance mechanisms and non-functional effector genes that spread within trial and intervention sites. This is of particular importance for non-localized gene drive projects, as the potential scale of intervention means that monitoring is expected to be more costly than research, development and deployment. Regarding monitoring needs for population replacement systems, lessons may be learned from experiences with Wolbachia-infected mosquitoes, and for population suppression systems, from experiences with releases of genetically sterile male mosquitoes. For population suppression systems, assessing monitoring requirements for tracking population size and detecting rare resistant alleles are priorities, while for population replacement systems, allele frequencies must be tracked, and pressing concerns include detection of gene drive alleles with non-functional effector genes, and resistance of pathogens to functional effector genes. For spread to unintended areas, open questions relate to the optimal density and placement of traps and frequency of sampling in order to detect gene drive alleles, drive-resistant alleles or non-functional effector genes while they can still be effectively managed. Invasive species management programs face similar questions, and lessons may be learned from these experiences. We explore these monitoring needs for gene drive mosquito projects progressing through the phases of pre-release, release and post-release.}, } @article {pmid35068955, year = {2021}, author = {Urfer, K and Spasojevic, T and Klopfstein, S and Baur, H and Lasut, L and Kropf, C}, title = {Incongruent molecular and morphological variation in the crab spider Synemaglobosum (Araneae, Thomisidae) in Europe.}, journal = {ZooKeys}, volume = {1078}, number = {}, pages = {107-134}, pmid = {35068955}, issn = {1313-2989}, abstract = {Establishing species boundaries is one of the challenges taxonomists around the world have been tackling for centuries. The relation between intraspecific and interspecific variability is still under discussion and in many taxa it remains understudied. Here the hypothesis of single versus multiple species of the crab spider Synemaglobosum (Fabricius) is tested. The wide distribution range as well as its high morphological variability makes this species an interesting candidate for re-evaluation using an integrative approach. This study combines information from barcoding, phylogenetic reconstruction based on mitochondrial CO1 and ITS2 of more than 60 specimens collected over a wide range of European localities, and morphology. The findings show deep clades with up to 6% mean pairwise distance in the CO1 barcode without any biogeographical pattern. The nuclear ITS2 gene did not support the CO1 clades. Morphological assessment of somatic and genital characters in males and females and a morphometric analysis of the male palp uncovered high intraspecific variation that does not match the CO1 or ITS2 phylogenies or biogeography either. Screening for endosymbiotic Wolbachia bacteria was conducted and only a single infected specimen was found. Several scenarios might explain these inconsistent patterns. While the deep divergences in the barcoding marker might suggest cryptic or ongoing speciation or geographical isolation in the past, the lack of congruent variation in the nuclear ITS2 gene or the studied morphological character systems, especially the male palp, indicates that S.globosum might simply be highly polymorphic both in terms of its mtDNA and morphology. Therefore, more data on ecology and behaviour and full genome sequences are necessary to ultimately resolve this taxonomically intriguing case.}, } @article {pmid35068604, year = {2022}, author = {Sureshan, SC and Mohideen, HS and Nair, TS}, title = {Gut Metagenomic Profiling of Gossypol Induced Oxycarenus laetus (Hemiptera: Lygaeidae) Reveals Gossypol Tolerating Bacterial Species.}, journal = {Indian journal of microbiology}, volume = {62}, number = {1}, pages = {54-60}, pmid = {35068604}, issn = {0046-8991}, abstract = {UNLABELLED: Oxycarenus laetus is a cotton pest that primarily feeds on seeds that are rich in gossypol. Though gossypol is toxic to general herbivores, O. laetus does not show such complications and instead grows and reproduces well on cotton plants compared to its other hosts. In this study, we have fed O. laetus with natural and induced gossypol-based diets to explore the difference in its gut microbiota. We performed NGS 16S rRNA amplicon sequencing on the Illumina MiSeq platform and analyzed the data using the QIIME2 pipeline supplemented with Greengenes and EZBioCloud reference databases. We also used culture-based methods to identify a few abundant gut bacteria present in O. laetus. Enterococcus faecalis, Wolbachia bourtzisii, Wolbachia pipientis, Corynebacterium glyciniphilum, Staphylococcus sciuri, and Kocuria rosea were some of the major species that formed the core gut microbiome of O. laetus. We have also observed that some species were present only in the sample with the highest concentration of gossypol, signifying that they might have the potential to degrade gossypol.

SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00964-0.}, } @article {pmid35067743, year = {2022}, author = {Lozano-Sardaneta, YN and Jacobo-Olvera, E and Ruiz-Tovar, K and Sánchez-Montes, S and Rodríguez-Rojas, JJ and Fernández-Figueroa, EA and Roldán-Fernández, SG and Rodriguez-Martinez, LM and Dzul-Manzanilla, F and Correa-Morales, F and Treviño-Garza, N and Díaz-Albíter, HM and Zwetsch, A and Valadas, SYOB and Nilce-Silveira, A and Becker, I and Huerta, H}, title = {Detection of Wolbachia and Leishmania DNA in sand flies (Diptera: Psychodidae, Phlebotominae) from a focus of cutaneous leishmaniasis in Tabasco, Mexico.}, journal = {Parasitology research}, volume = {121}, number = {2}, pages = {513-520}, pmid = {35067743}, issn = {1432-1955}, support = {6682//consejo nacional de ciencia y tecnología/ ; IG201221//programa de apoyo a proyectos de investigación e innovación tecnológica (papiit)/ ; }, mesh = {Animals ; DNA ; Female ; Insect Vectors ; *Leishmania/genetics ; *Leishmaniasis, Cutaneous ; Mexico ; *Psychodidae/genetics ; *Wolbachia/genetics ; }, abstract = {Phlebotomine sand flies are the main vectors of Leishmania genus species worldwide; therefore, the detection of some reproductive parasites, such as Wolbachia, has been considered a possible strategy for biological control. In Mexico, leishmaniasis cases have been recorded in 25 states, yet only two sand fly species have been related to Wolbachia spp. Although the state of Tabasco has a high number of leishmaniasis cases, only few studies have been done on sand fly species. The aim of this study was to analyze the diversity of sand fly species and to detect Wolbachia spp. and/or Leishmania spp. in the captured specimens. Sand flies were collected at the locality of Huimango, Tabasco, Mexico, during October 2019, using nine light traps (CDC) and two Shannon traps per night. The specimens were identified and females were analyzed by PCR for the DNA detection for pathogens. A total of 193 sand fly specimens belonging to five species were morphologically identified. Pintomyia ovallesi was the most abundant species (76.84%), followed by Micropygomyia cayennensis (6.40%). Furthermore, first records of four sand fly species were established for the state of Tabasco, thereby increasing the species richness in the state from four to eight. We observed a natural infection rate of 9.7% (10/103) for Leishmania and 0.91% (1/103) for Wolbachia. The importance of conducting entomological surveys in endemic areas of leishmaniasis in Mexico is highlighted, to determine whether other sand fly species may be potential vectors of Leishmania spp., and if some Wolbachia strains could be relevant for the control of leishmaniasis.}, } @article {pmid35066589, year = {2022}, author = {Hidayanti, AK and Gazali, A and Tagami, Y}, title = {Effect of Quorum Sensing Inducers and Inhibitors on Cytoplasmic Incompatibility Induced by Wolbachia (Rickettsiales: Anaplasmataceae) in American Serpentine Leafminer (Diptera: Agromyzidae): Potential Tool for the Incompatible Insect Technique.}, journal = {Journal of insect science (Online)}, volume = {22}, number = {1}, pages = {}, pmid = {35066589}, issn = {1536-2442}, mesh = {Animals ; *Diptera/microbiology ; Ovum ; *Pest Control, Biological ; *Quorum Sensing ; *Wolbachia ; }, abstract = {Agricultural crops around the world are attacked by approximately 3,000-10,000 species of pest insect. There is increasing interest in resolving this problem using environmentally friendly approaches. Wolbachia (Hertig), an insect endosymbiont, can modulate host reproduction and offspring sex through cytoplasmic incompatibility (CI). The incompatible insect technique (IIT) based on CI-Wolbachia is a promising biological control method. Previous studies have reported an association between CI and Wolbachia density, which may involve a quorum sensing (QS) mechanism. In this study, we investigated the effect of manipulating QS in Wolbachia using several chemicals including 3O-C12-HSL; C2HSL; spermidine (QS inducers), 4-phenylbutanoyl; and 4-NPO (QS inhibitors) on American serpentine leafminer (Liriomyza trifolii [Burgess]), an agricultural pest. The results showed that inducing QS with 3O-C12-HSL decreased the proportion of hatched eggs and increased Wolbachia density, whereas QS inhibition with 4-phenylbutanoyl had the opposite effects. Thus, manipulating QS in Wolbachia can alter cell density and the proportion of hatched eggs in the host L. trifolii, thereby reducing the number of insect progeny. These findings provide evidence supporting the potential efficacy of the IIT based on CI-Wolbachia for the environmentally friendly control of insect pest populations.}, } @article {pmid35064627, year = {2022}, author = {Li, TP and Zhou, CY and Gong, JT and Xi, Z and Hong, XY}, title = {Recently introduced Wolbachia reduces bacterial species richness and reshapes bacterial community structure in Nilaparvata lugens.}, journal = {Pest management science}, volume = {78}, number = {5}, pages = {1881-1894}, doi = {10.1002/ps.6806}, pmid = {35064627}, issn = {1526-4998}, support = {2016YFC1201200//National Key Research and Development Program of China/ ; 32020103011//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Bacteria ; *Hemiptera/microbiology ; Humans ; *Microbiota ; Nymph/microbiology ; *Wolbachia ; }, abstract = {BACKGROUND: Wolbachia has been developed as an effective tool to suppress insect pests and arbovirus transmission. Recently, the brown planthopper Nilaparvata lugens, a serious agricultural pest, has been successfully transinfected with Wolbachia wStri strain from Laodelphax striatellus. However, before conducting the field experiments, the impacts of wStri on the bacterial microbiota in N. lugens and how it differs from native Wolbachia wLug strain have not been clarified.

RESULTS: Here, we found that wStri reduced bacterial diversity and shaped bacterial community structure more than wLug in both developmental stage and different adult tissues. Overall, the relative abundance of Wolbachia was negatively correlated with bacterial diversity, but the bacterial diversity gradually decreased only when the relative abundance of Wolbachia was higher than 60%. Further analysis found that wStri reduced species richness of other bacteria but not their evenness. wStri infection also affected many bacterial functions (e.g., amino acid metabolism & signaling and cellular processes) in the developmental stages, with a stronger effect than wLug in nymphs. Moreover, although Wolbachia occupied a high relative abundance in infected individuals, Acinetobacter was consistently a core part of microbiome.

CONCLUSION: These results showed the significant impacts of recently introduced wStri on bacterial microbiota in N. lugens, with the effects differing from native wLug. This study will aid in understanding the relationship between Wolbachia, its host and the host's microbiota, and provide a reference for future field experiments.}, } @article {pmid35059821, year = {2023}, author = {Mancini, E and Sabatelli, S and Hu, Y and Frasca, S and Di Giulio, A and Audisio, P and Brown, CD and Russell, JA and Trizzino, M}, title = {Uncovering Active Bacterial Symbionts in Three Species of Pollen-feeding Beetles (Nitidulidae: Meligethinae).}, journal = {Microbial ecology}, volume = {85}, number = {1}, pages = {335-339}, pmid = {35059821}, issn = {1432-184X}, support = {1442144//national science foundation/ ; RP120172B8C07DF1//sapienza università di roma/ ; CAL/2016//università degli studi roma tre/ ; }, mesh = {Animals ; *Coleoptera ; RNA, Ribosomal, 16S/genetics ; Insecta ; Pollen ; Plants ; }, abstract = {Microbial symbionts enable many phytophagous insects to specialize on plant-based diets through a range of metabolic services. Pollen comprises one-plant tissue consumed by such herbivores. While rich in lipids and proteins, its nutrient content is often imbalanced and difficult-to-access due to a digestibly recalcitrant cell wall. Pollen quality can be further degraded by harmful allelochemicals. To identify microbes that may aid in palynivory, we performed cDNA-based 16S rRNA metabarcoding on three related pollen beetles (Nitidulidae: Meligethinae) exhibiting different dietary breadths: Brassicogethes aeneus, B. matronalis, and Meligethes atratus. Nine bacterial symbionts (i.e., 97% OTUs) exhibited high metabolic activity during active feeding. Subsequent PCR surveys revealed varying prevalence of those from three Rickettsialles genera-Lariskella, Rickettsia, and Wolbachia-within beetle populations. Our findings lay the groundwork for future studies on the influence of phylogeny and diet on palynivorous insect microbiomes, and roles of symbionts in the use of challenging diets.}, } @article {pmid35055992, year = {2021}, author = {Efon Ekangouo, A and Nana Djeunga, HC and Sempere, G and Kamgno, J and Njiokou, F and Moundipa Fewou, P and Geiger, A}, title = {Bacteriome Diversity of Blackflies' Gut and Association with Onchocerca volvulus, the Causative Agent of Onchocerciasis in Mbam Valley (Center Region, Cameroon).}, journal = {Pathogens (Basel, Switzerland)}, volume = {11}, number = {1}, pages = {}, pmid = {35055992}, issn = {2076-0817}, abstract = {Vector control using larvicides is the main alternative strategy to address limits of preventive chemotherapy using ivermectin for the control of onchocerciasis. However, it remains substantially limited by implementation difficulties, ecological concerns and the resistance of vector populations. Therefore, efficient and environmentally safe alternative control strategies are still needed. This study explores the composition of the blackfly bacteriome and its variability in the presence of Onchocerca volvulus infection, in order to determine their potential as a novel vector control-based approach to fight onchocerciasis. An entomological survey of a collection of samples was performed in the Bafia health district, a historical endemic focus for onchocerciasis in Cameroon. A total of 1270 blackflies were dissected and the infection rate was 10.1%, indicative of ongoing transmission of onchocerciasis in the surveyed communities. Sequencing process of blackflies' gut DNA for bacteria screening revealed 14 phyla and 123 genera, highlighting the diversity of gut blackflies bacterial communities. Eight bacteria formed the core of blackfly bacteriome and Wolbachia was the predominant genus with 73.4% of relative abundance of blackflies' gut bacterial communities. Acidomonas and Roseanomas genera were significantly abundant among infected blackflies (p = 0.01), whereas other genera such as Brevibacterium and Fructobacillus were associated with the absence of infection (p = 0.0009). Differences in gut bacterial distribution of blackflies according to their infection status by the parasite suggest a causal relationship between the bacteriome composition and the onset of blackflies' infection by O. volvulus or vice versa. Blackfly native bacteria are then potentially involved in infection by O. volvulus, either by facilitating or preventing the parasite infestation of the vector. These bacteria represent an interesting potential as a biological tool/target for a novel approach of vector control to fight onchocerciasis.}, } @article {pmid35055852, year = {2021}, author = {Du, S and Ye, F and Wang, Q and Liang, Y and Wan, W and Guo, J and Liu, W}, title = {Multiple Data Demonstrate That Bacteria Regulating Reproduction Could Be Not the Cause for the Thelytoky of Diglyphus&nbsp;wani (Hymenoptera: Eulophidae).}, journal = {Insects}, volume = {13}, number = {1}, pages = {}, pmid = {35055852}, issn = {2075-4450}, support = {Grant No. 31772236 and No. 31972344//the National Natural Science Foundation of China/ ; Grant No. caascx-2017-2022-IAS//the Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences/ ; Grant No. 2021YFC2600400//the National Key R&D Program of China/ ; }, abstract = {In Hymenoptera parasitoids, the reproductive mode is arrhenotoky, while a few species reproduce by thelytoky. The thelytoky of Hymenoptera parasitoids is generally genetically determined by the parasitoids themselves or induced by bacteria, including Wolbachia, Cardinium, and Rickettsia. Diglyphus&nbsp;wani (Hymenoptera: Eulophidae), a recently reported thelytokous species is a main parasitoid attacking agromyzid leafminers. To assess whether endosymbionts induce thelytoky in D.&nbsp;wani, we performed universal PCR detection and sequenced the V3-V4 region of 16S ribosomal RNA gene. In addition, bacteria were removed through high-temperature and antibiotic treatments, and the localized bacteria were detected using FISH. Based on general PCR detection, Wolbachia, Cardinium, Rickettsia, Arsenophonus, Spiroplasma, and Microsporidia were absent in laboratory and field individuals of thelytokous D.&nbsp;wani. Furthermore, 16S rRNA gene sequencing revealed that the dominant endosymbionts in thelytokous D.&nbsp;wani were not reproductive manipulators. High-temperature and antibiotic treatment for five consecutive generations cannot reverse the thelytokous pattern of D.&nbsp;wani, and no male offspring were produced. Moreover, no bacterial spots were found in the ovaries of D.&nbsp;wani. Thus, it is considered that the thelytoky of D. wani does not result in the presence of endosymbionts. This species is thus the second reported eulophid parasitoid whose thelytoky appears not to be associated with endosymbionts.}, } @article {pmid35055844, year = {2021}, author = {Aryaprema, VS and Qualls, WA and Dobson, KL and Dobson, SL and Xue, RD}, title = {The Effects of Boric Acid Sugar Bait on Wolbachia Trans-Infected Male Aedes albopictus (ZAP Males[®]) in Laboratory Conditions.}, journal = {Insects}, volume = {13}, number = {1}, pages = {}, pmid = {35055844}, issn = {2075-4450}, abstract = {The field release of Wolbachia trans-infected male mosquitoes, as well as the use of toxic sugar baits, is a novel and promising candidate technique for integrated mosquito management programs. However, the methods of action of the two techniques may not be complementary, because the Wolbachia method releases mosquitoes into the environment expecting a wild population reduction in subsequent generations while the toxic baits are intended to reduce the wild population by killing mosquitoes. This laboratory study was conducted to evaluate the effectiveness of boric acid toxic sugar baits on Wolbachia trans-infected male Aedes albopictus, relative to wild-type Ae. albopictus males. Wolbachia trans-infected (ZAP male[®]) and the wild-type Ae. albopictus males were exposed separately to 1% boric acid in a 10% sucrose solution in BugDorms. In the control test, the two groups were exposed to 10% sucrose solution without boric acid. Percent mortalities were counted for 24 h, 48 h and 72 h post exposure periods. The results show that 1% boric acid toxic sugar bait can effectively kill ZAP males under laboratory conditions, and the effectiveness was significantly higher after 24 h and 48 h, compared to wild-type male Ae. albopictus. This finding will help in planning and coordinating integrated mosquito management programs, including both Wolbachia trans-infected mosquito releases and the use of toxic sugar baits against Ae. albopictus.}, } @article {pmid35055035, year = {2022}, author = {Ilinsky, Y and Demenkova, M and Bykov, R and Bugrov, A}, title = {Narrow Genetic Diversity of Wolbachia Symbionts in Acrididae Grasshopper Hosts (Insecta, Orthoptera).}, journal = {International journal of molecular sciences}, volume = {23}, number = {2}, pages = {}, pmid = {35055035}, issn = {1422-0067}, support = {19-04-00983//Russian Foundation for Basic Research/ ; 121031800061-7//Ministry of Science and Higher Education (Russian Federation)/ ; }, mesh = {Animals ; Gene Transfer, Horizontal ; *Genetic Variation ; Grasshoppers/*microbiology ; Insecta/microbiology ; Multilocus Sequence Typing ; Phylogeny ; Symbiosis ; Wolbachia/*classification/*genetics ; }, abstract = {Bacteria of the Wolbachia genus are maternally inherited symbionts of Nematoda and numerous Arthropoda hosts. There are approximately 20 lineages of Wolbachia, which are called supergroups, and they are designated alphabetically. Wolbachia strains of the supergroups A and B are predominant in arthropods, especially in insects, and supergroup F seems to rank third. Host taxa have been studied very unevenly for Wolbachia symbionts, and here, we turn to one of largely unexplored insect families: Acrididae. On the basis of five genes subject to multilocus sequence typing, we investigated the incidence and genetic diversity of Wolbachia in 41 species belonging three subfamilies (Gomphocerinae, Oedipodinae, and Podisminae) collected in Turkey, Kazakhstan, Tajikistan, Russia, and Japan, making 501 specimens in total. Our results revealed a high incidence and very narrow genetic diversity of Wolbachia. Although only the strains belonging to supergroups A and B are commonly present in present, the Acrididae hosts here proved to be infected with supergroups B and F without A-supergroup variants. The only trace of an A-supergroup lineage was noted in one case of an inter-supergroup recombinant haplotype, where the ftsZ gene came from supergroup A, and the others from supergroup B. Variation in the Wolbachia haplotypes in Acrididae hosts within supergroups B and F was extremely low. A comprehensive genetic analysis of Wolbachia diversity confirmed specific features of the Wolbachia allelic set in Acrididae hosts. This result can help to elucidate the crucial issue of Wolbachia biology: the route(s) and mechanism(s) of Wolbachia horizontal transmission.}, } @article {pmid35030199, year = {2022}, author = {Aikawa, T and Maehara, N and Ichihara, Y and Masuya, H and Nakamura, K and Anbutsu, H}, title = {Cytoplasmic incompatibility in the semivoltine longicorn beetle Acalolepta fraudatrix (Coleoptera: Cerambycidae) double infected with Wolbachia.}, journal = {PloS one}, volume = {17}, number = {1}, pages = {e0261928}, pmid = {35030199}, issn = {1932-6203}, mesh = {Animals ; Coleoptera/*microbiology/ultrastructure ; DNA, Bacterial/*genetics ; Asia, Eastern ; Female ; Male ; RNA, Bacterial/*genetics ; RNA, Ribosomal, 16S/*genetics ; *Wolbachia/classification/genetics/ultrastructure ; }, abstract = {Wolbachia are obligatory endosymbiotic α-proteobacteria found in many arthropods. They are maternally inherited, and can induce reproductive alterations in the hosts. Despite considerable recent progress in studies on the associations between Wolbachia and various taxonomic groups of insects, none of the researches have revealed the effects of Wolbachia on longicorn beetles as the host insect. Acalolepta fraudatrix is a forest longicorn beetle that is distributed in East Asia. In this study, the relationship between Wolbachia and A. fraudatrix was investigated. Out of two populations of A. fraudatrix screened for Wolbachia using the genes ftsZ, wsp, and 16S rRNA, only one of the populations showed detection of all three genes indicating the presence of Wolbachia. Electron microscopy and fluorescent in situ hybridization also confirmed that the A. fraudatrix population was infected with Wolbachia. Sequencing the wsp genes derived from single insects revealed that two strains of Wolbachia coexisted in the insects based on the detection of two different sequences of the wsp gene. We designated these strains as wFra1 and wFra2. The bacterial titers of wFra1 were nearly 2-fold and 3-fold higher than wFra2 in the testes and ovaries, respectively. The two strains of Wolbachia in the insects were completely eliminated by rearing the insects on artificial diets containing 1% concentration of tetracycline for 1 generation. Reciprocal crosses between Wolbachia-infected and Wolbachia-uninfected A. fraudatrix demonstrated that only eggs produced by the crosses between Wolbachia-infected males and Wolbachia-uninfected females did not hatch, indicating that Wolbachia infecting A. fraudatrix causes cytoplasmic incompatibility in the host insect. This is the first report showing the effect of Wolbachia on reproductive function in a longicorn beetle, A. fraudatrix.}, } @article {pmid35029964, year = {2022}, author = {Rybnicky, GA and Dixon, RA and Kuhn, RM and Karim, AS and Jewett, MC}, title = {Development of a Freeze-Dried CRISPR-Cas12 Sensor for Detecting Wolbachia in the Secondary Science Classroom.}, journal = {ACS synthetic biology}, volume = {11}, number = {2}, pages = {835-842}, doi = {10.1021/acssynbio.1c00503}, pmid = {35029964}, issn = {2161-5063}, mesh = {Biotechnology ; CRISPR-Cas Systems/genetics ; Gene Editing ; Genetic Engineering ; Humans ; Synthetic Biology/education ; *Wolbachia/genetics ; }, abstract = {Training the future synthetic biology workforce requires the opportunity for students to be exposed to biotechnology concepts and activities in secondary education. Detecting Wolbachia bacteria in arthropods using polymerase chain reaction (PCR) has become a common way for secondary students to investigate and apply recombinant DNA technology in the science classroom. Despite this important activity, cutting-edge biotechnologies such as clustered regularly interspaced short palindromic repeat (CRISPR)-based diagnostics have yet to be widely implemented in the classroom. To address this gap, we present a freeze-dried CRISPR-Cas12 sensing reaction to complement traditional recombinant DNA technology education and teach synthetic biology concepts. The reactions accurately detect Wolbachia from arthropod-derived PCR samples in under 2 h and can be stored at room temperature for over a month without appreciable degradation. The reactions are easy-to-use and cost less than $40 to implement for a classroom of 22 students including the cost of reusable equipment. We see these freeze-dried CRISPR-Cas12 reactions as an accessible way to incorporate synthetic biology education into the existing biology curriculum, which will expand biology educational opportunities in science, technology, engineering, and mathematics.}, } @article {pmid35027704, year = {2022}, author = {Hofer, U}, title = {Wolbachia likes it hot.}, journal = {Nature reviews. Microbiology}, volume = {20}, number = {3}, pages = {126}, pmid = {35027704}, issn = {1740-1534}, mesh = {*Aedes ; Animals ; Anti-Bacterial Agents ; *Wolbachia/genetics ; }, } @article {pmid35019223, year = {2022}, author = {Richardson, KM and Schiffer, M and Ross, PA and Thia, JA and Hoffmann, AA}, title = {Characterization of the first Wolbachia from the genus Scaptodrosophila, a male-killer from the rainforest species S. claytoni.}, journal = {Insect science}, volume = {29}, number = {5}, pages = {1401-1413}, doi = {10.1111/1744-7917.13000}, pmid = {35019223}, issn = {1744-7917}, mesh = {Animals ; Drosophila/genetics ; Female ; Male ; Multilocus Sequence Typing ; Phylogeny ; Rainforest ; Tetracyclines ; *Wolbachia/genetics ; }, abstract = {The Scaptodrosophila genus represents a large group of drosophilids with a worldwide distribution and a predominance of species in Australia, but there is little information on the presence and impacts of Wolbachia endosymbionts in this group. Here we describe the first Wolbachia infection from this group, wClay isolated from Scaptodrosophila claytoni (van Klinken), a species from the east coast of Australia. The infection is polymorphic in natural populations, occurring at a frequency of around 6%-10%. wClay causes male killing, producing female-biased lines; most lines showed 100% male killing, though in 1 line it was <80%. The lines need to be maintained through the introduction of males unless the infection is removed by tetracycline treatment. wClay is transmitted at a high fidelity (98.6%) through the maternal lineage and has been stable in 2 laboratory lines across 24 generations, suggesting it is likely to persist in populations. The infection has not been previously described but is closely related to the male-killing Wolbachia recently described from Drosophila pandora based on multilocus sequence typing and the wsp gene. Male-killing Wolbachia are likely to be common in drosophilids but remain difficult to detect because the infections can often be at a low frequency.}, } @article {pmid35015769, year = {2022}, author = {Liu, WL and Yu, HY and Chen, YX and Chen, BY and Leaw, SN and Lin, CH and Su, MP and Tsai, LS and Chen, Y and Shiao, SH and Xi, Z and Jang, AC and Chen, CH}, title = {Lab-scale characterization and semi-field trials of Wolbachia Strain wAlbB in a Taiwan Wolbachia introgressed Ae. aegypti strain.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {1}, pages = {e0010084}, pmid = {35015769}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Biological Control Agents/administration & dosage ; Dengue/*prevention & control/transmission ; Dengue Virus/isolation & purification ; Female ; Male ; Mosquito Control/*methods ; Mosquito Vectors/virology ; Pest Control, Biological/*methods ; Taiwan ; Wolbachia/classification/*metabolism ; Zygote/microbiology ; }, abstract = {Dengue fever is one of the most severe viral diseases transmitted by Aedes mosquitoes, with traditional approaches of disease control proving insufficient to prevent significant disease burden. Release of Wolbachia-transinfected mosquitoes offers a promising alternative control methodologies; Wolbachia-transinfected female Aedes aegypti demonstrate reduced dengue virus transmission, whilst Wolbachia-transinfected males cause zygotic lethality when crossed with uninfected females, providing a method for suppressing mosquito populations. Although highly promising, the delicate nature of population control strategies and differences between local species populations means that controlled releases of Wolbachia-transinfected mosquitoes cannot be performed without extensive testing on specific local Ae. aegypti populations. In order to investigate the potential for using Wolbachia to suppress local Ae. aegypti populations in Taiwan, we performed lab-based and semi-field fitness trials. We first transinfected the Wolbachia strain wAlbB into a local Ae. aegypti population (wAlbB-Tw) and found no significant changes in lifespan, fecundity and fertility when compared to controls. In the laboratory, we found that as the proportion of released male mosquitoes carrying Wolbachia was increased, population suppression could reach up to 100%. Equivalent experiments in semi-field experiments found suppression rates of up to 70%. The release of different ratios of wAlbB-Tw males in the semi-field system provided an estimate of the optimal size of male releases. Our results indicate that wAlbB-Tw has significant potential for use in vector control strategies aimed at Ae. aegypti population suppression in Taiwan. Open field release trials are now necessary to confirm that wAlbB-Tw mediated suppression is feasible in natural environments.}, } @article {pmid35013476, year = {2022}, author = {Towett-Kirui, S and Morrow, JL and Riegler, M}, title = {Substantial rearrangements, single nucleotide frameshift deletion and low diversity in mitogenome of Wolbachia-infected strepsipteran endoparasitoid in comparison to its tephritid hosts.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {477}, pmid = {35013476}, issn = {2045-2322}, support = {2018 E.A. Southee Award//Hawkesbury Foundation/ ; IC150100026//Australian Research Council/ ; }, mesh = {Animals ; Australia ; Frameshift Mutation ; Gene Rearrangement ; *Genome, Insect ; *Genome, Mitochondrial ; Insect Proteins/genetics ; Sequence Deletion ; Tephritidae/classification/*genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Insect mitogenome organisation is highly conserved, yet, some insects, especially with parasitic life cycles, have rearranged mitogenomes. Furthermore, intraspecific mitochondrial diversity can be reduced by fitness-affecting bacterial endosymbionts like Wolbachia due to their maternal coinheritance with mitochondria. We have sequenced mitogenomes of the Wolbachia-infected endoparasitoid Dipterophagus daci (Strepsiptera: Halictophagidae) and four of its 22 known tephritid fruit fly host species using total genomic extracts of parasitised flies collected across > 700 km in Australia. This halictophagid mitogenome revealed extensive rearrangements relative to the four fly mitogenomes which exhibited the ancestral insect mitogenome pattern. Compared to the only four available other strepsipteran mitogenomes, the D. daci mitogenome had additional transpositions of one rRNA and two tRNA genes, and a single nucleotide frameshift deletion in nad5 requiring translational frameshifting or, alternatively, resulting in a large protein truncation. Dipterophagus daci displays an almost completely endoparasitic life cycle when compared to Strepsiptera that have maintained the ancestral state of free-living adults. Our results support the hypothesis that the transition to extreme endoparasitism evolved together with increased levels of mitogenome changes. Furthermore, intraspecific mitogenome diversity was substantially smaller in D. daci than the parasitised flies suggesting Wolbachia reduced mitochondrial diversity because of a role in D. daci fitness.}, } @article {pmid35006317, year = {2022}, author = {Schlabe, S and Korir, P and Lämmer, C and Landmann, F and Dubben, B and Koschel, M and Albers, A and Debrah, LB and Debrah, AY and Hübner, MP and Pfarr, K and Klarmann-Schulz, U and Hoerauf, A}, title = {A qPCR to quantify Wolbachia from few Onchocerca volvulus microfilariae as a surrogate for adult worm histology in clinical trials of antiwolbachial drugs.}, journal = {Parasitology research}, volume = {121}, number = {4}, pages = {1199-1206}, pmid = {35006317}, issn = {1432-1955}, support = {OPPGH5342//Bill and Melinda Gates Foundation/ ; TI 07.001_Schlabe_00//Deutsches Zentrum für Infektionsforschung, Clinical Leave Stipend/ ; TI 07.001_Schlabe_01//Deutsches Zentrum für Infektionsforschung, Clinical Leave Stipend/ ; TI 03.907//Deutsches Zentrum für Infektionsforschung/ ; EXC2151 - 390873048//Deutsche Forschungsgemeinschaft/ ; }, mesh = {Animals ; Humans ; *Filarioidea ; Microfilariae ; Onchocerca ; *Onchocerca volvulus/genetics ; Reproducibility of Results ; *Wolbachia/drug effects/genetics ; }, abstract = {The filarial nematode Onchocerca volvulus causes onchocerciasis (river blindness), a neglected tropical disease affecting 21 million people, mostly in Sub-Saharan Africa. Targeting the endosymbiont Wolbachia with antibiotics leads to permanent sterilization and killing of adult worms. The gold standard to assess Wolbachia depletion is the histological examination of adult worms in nodules beginning at 6 months post-treatment. However, nodules can only be used once, limiting the time points to monitor Wolbachia depletion. A diagnostic to longitudinally monitor Wolbachia depletion from microfilariae (MF) at more frequent intervals < 6 months post-treatment would accelerate clinical trials of antiwolbachials. We developed a TaqMan qPCR amplifying the single-copy gene wOvftsZ to quantify Wolbachia from as few as one MF that had migrated from skin biopsies and compared quantification using circular and linearized plasmids or synthetic dsDNA (gBlock®). qPCR for MF from the rodent nematode Litomosoides sigmodontis was used to support the reproducibility and validate the principle. The qPCR using as few as 2 MF from O. volvulus and L. sigmodontis reproducibly quantified Wolbachia. Use of a linearized plasmid standard or synthesized dsDNA resulted in numbers of Wolbachia/MF congruent with biologically plausible estimates in O. volvulus and L. sigmodontis MF. The qPCR assay yielded a median of 48.8 (range 1.5-280.5) Wolbachia/O. volvulus MF. The qPCR is a sensitive tool for quantifying Wolbachia in a few MF from skin biopsies and allows for establishing the qPCR as a surrogate parameter for monitoring Wolbachia depletion in adult worms of new antiwolbachial candidates.}, } @article {pmid35006065, year = {2022}, author = {Leitner, M and Etebari, K and Asgari, S}, title = {Transcriptional response of Wolbachia-transinfected Aedes aegypti mosquito cells to dengue virus at early stages of infection.}, journal = {The Journal of general virology}, volume = {103}, number = {1}, pages = {}, pmid = {35006065}, issn = {1465-2099}, mesh = {Aedes/microbiology/*virology ; Animals ; Dengue/*virology ; Dengue Virus/*genetics/physiology ; Host Microbial Interactions ; Humans ; Mosquito Vectors/microbiology/virology ; RNA, Long Noncoding ; Sumoylation ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Mosquito-borne flaviviruses are responsible for viral infections and represent a considerable public health burden. Aedes aegypti is the principal vector of dengue virus (DENV), therefore understanding the intrinsic virus-host interactions is vital, particularly in the presence of the endosymbiont Wolbachia, which blocks virus replication in mosquitoes. Here, we examined the transcriptional response of Wolbachia-transinfected Ae. aegypti Aag2 cells to DENV infection. We identified differentially expressed immune genes that play a key role in the activation of anti-viral defence such as the Toll and immune deficiency pathways. Further, genes encoding cytosine and N[6]-adenosine methyltransferases and SUMOylation, involved in post-transcriptional modifications, an antioxidant enzyme, and heat-shock response were up-regulated at the early stages of DENV infection and are reported here for the first time. Additionally, several long non-coding RNAs were among the differentially regulated genes. Our results provide insight into Wolbachia-transinfected Ae. aegypti's initial virus recognition and transcriptional response to DENV infection.}, } @article {pmid34996906, year = {2022}, author = {Neupane, S and Bonilla, SI and Manalo, AM and Pelz-Stelinski, KS}, title = {Complete de novo assembly of Wolbachia endosymbiont of Diaphorina citri Kuwayama (Hemiptera: Liviidae) using long-read genome sequencing.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {125}, pmid = {34996906}, issn = {2045-2322}, support = {D19AP00013//Defense Advanced Research Projects Agency/ ; D19AP00013//Defense Advanced Research Projects Agency/ ; D19AP00013//Defense Advanced Research Projects Agency/ ; D19AP00013//Defense Advanced Research Projects Agency/ ; }, mesh = {Animals ; Cell Line ; *Chromosomes, Bacterial ; DNA, Bacterial/*genetics ; DNA, Circular/*genetics ; *Genes, Bacterial ; *Genome, Bacterial ; Hemiptera/*microbiology ; Phylogeny ; Sequence Analysis, DNA ; Symbiosis ; *Whole Genome Sequencing ; Wolbachia/*genetics ; }, abstract = {Wolbachia, a gram-negative [Formula: see text]-proteobacterium, is an endosymbiont found in some arthropods and nematodes. Diaphorina citri Kuwayama, the vector of 'Candidatus Liberibacter asiaticus' (CLas), are naturally infected with a strain of Wolbachia (wDi), which has been shown to colocalize with the bacteria pathogens CLas, the pathogen associated with huanglongbing (HLB) disease of citrus. The relationship between wDi and CLas is poorly understood in part because the complete genome of wDi has not been available. Using high-quality long-read PacBio circular consensus sequences, we present the largest complete circular wDi genome among supergroup-B members. The assembled circular chromosome is 1.52 megabases with 95.7% genome completeness with contamination of 1.45%, as assessed by checkM. We identified Insertion Sequences (ISs) and prophage genes scattered throughout the genomes. The proteins were annotated using Pfam, eggNOG, and COG that assigned unique domains and functions. The wDi genome was compared with previously sequenced Wolbachia genomes using pangenome and phylogenetic analyses. The availability of a complete circular chromosome of wDi will facilitate understanding of its role within the insect vector, which may assist in developing tools for disease management. This information also provides a baseline for understanding phylogenetic relationships among Wolbachia of other insect vectors.}, } @article {pmid34996376, year = {2022}, author = {Nakabachi, A and Inoue, H and Hirose, Y}, title = {Microbiome analyses of 12 psyllid species of the family Psyllidae identified various bacteria including Fukatsuia and Serratia symbiotica, known as secondary symbionts of aphids.}, journal = {BMC microbiology}, volume = {22}, number = {1}, pages = {15}, pmid = {34996376}, issn = {1471-2180}, mesh = {Animals ; Aphids/microbiology ; Gammaproteobacteria/classification/genetics/*isolation & purification ; Hemiptera/classification/*microbiology ; Liberibacter/classification/genetics/isolation & purification ; *Microbiota ; Phylogeny ; Plant Diseases/microbiology/parasitology ; Rickettsia/classification/genetics/isolation & purification ; Serratia/classification/genetics/isolation & purification ; Symbiosis ; Wolbachia/classification/genetics/isolation & purification ; }, abstract = {BACKGROUND: Psyllids (Hemiptera: Psylloidea) comprise a group of plant sap-sucking insects that includes important agricultural pests. They have close associations not only with plant pathogens, but also with various microbes, including obligate mutualists and facultative symbionts. Recent studies are revealing that interactions among such bacterial populations are important for psyllid biology and host plant pathology. In the present study, to obtain further insight into the ecological and evolutionary behaviors of bacteria in Psylloidea, we analyzed the microbiomes of 12 psyllid species belonging to the family Psyllidae (11 from Psyllinae and one from Macrocorsinae), using high-throughput amplicon sequencing of the 16S rRNA gene.

RESULTS: The analysis showed that all 12 psyllids have the primary symbiont, Candidatus Carsonella ruddii (Gammaproteobacteria: Oceanospirillales), and at least one secondary symbiont. The majority of the secondary symbionts were gammaproteobacteria, especially those of the family Enterobacteriaceae (order: Enterobacteriales). Among them, symbionts belonging to "endosymbionts3", which is a genus-level monophyletic group assigned by the SILVA rRNA database, were the most prevalent and were found in 9 of 11 Psyllinae species. Ca. Fukatsuia symbiotica and Serratia symbiotica, which were recognized only as secondary symbionts of aphids, were also identified. In addition to other Enterobacteriaceae bacteria, including Arsenophonus, Sodalis, and "endosymbionts2", which is another genus-level clade, Pseudomonas (Pseudomonadales: Pseudomonadaceae) and Diplorickettsia (Diplorickettsiales: Diplorickettsiaceae) were identified. Regarding Alphaproteobacteria, the potential plant pathogen Ca. Liberibacter europaeus (Rhizobiales: Rhizobiaceae) was detected for the first time in Anomoneura mori (Psyllinae), a mulberry pest. Wolbachia (Rickettsiales: Anaplasmataceae) and Rickettsia (Rickettsiales: Rickettsiaceae), plausible host reproduction manipulators that are potential tools to control pest insects, were also detected.

CONCLUSIONS: The present study identified various bacterial symbionts including previously unexpected lineages in psyllids, suggesting considerable interspecific transfer of arthropod symbionts. The findings provide deeper insights into the evolution of interactions among insects, bacteria, and plants, which may be exploited to facilitate the control of pest psyllids in the future.}, } @article {pmid34969947, year = {2021}, author = {Ghanavi, HR and Twort, VG and Duplouy, A}, title = {Exploring bycatch diversity of organisms in whole genome sequencing of Erebidae moths (Lepidoptera).}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {24499}, pmid = {34969947}, issn = {2045-2322}, mesh = {Animals ; Bacteria/genetics ; Genome, Insect ; Lepidoptera/genetics/physiology ; Moths/*genetics/*microbiology/physiology ; *Symbiosis ; Whole Genome Sequencing ; Wolbachia/genetics/physiology ; }, abstract = {Models estimate that up to 80% of all butterfly and moth species host vertically transmitted endosymbiotic microorganisms, which can affect the host fitness, metabolism, reproduction, population dynamics, and genetic diversity, among others. The supporting empirical data are however currently highly biased towards the generally more colourful butterflies, and include less information about moths. Additionally, studies of symbiotic partners of Lepidoptera predominantly focus on the common bacterium Wolbachia pipientis, while infections by other inherited microbial partners have more rarely been investigated. Here, we mine the whole genome sequence data of 47 species of Erebidae moths, with the aims to both inform on the diversity of symbionts potentially associated with this Lepidoptera group, and discuss the potential of metagenomic approaches to inform on host associated microbiome diversity. Based on the result of Kraken2 and MetaPhlAn2 analyses, we found clear evidence of the presence of Wolbachia in four species. Our result also suggests the presence of three other bacterial symbionts (Burkholderia spp., Sodalis spp. and Arsenophonus spp.) in three other moth species. Additionally, we recovered genomic material from bracovirus in about half of our samples. The detection of the latter, usually found in mutualistic association to braconid parasitoid wasps, may inform on host-parasite interactions that take place in the natural habitat of the Erebidae moths, suggesting either contamination with material from species of the host community network, or horizontal transfer of members of the microbiome between interacting species.}, } @article {pmid34965003, year = {2022}, author = {Xu, P and Rice, A and Li, T and Wang, J and Yang, X and Yuan, H and Graham, RI and Wilson, K}, title = {Partiti-like viruses from African armyworm increase larval and pupal mortality of a novel host: the Egyptian cotton leafworm.}, journal = {Pest management science}, volume = {78}, number = {4}, pages = {1529-1537}, doi = {10.1002/ps.6771}, pmid = {34965003}, issn = {1526-4998}, support = {//Newton international fellowship (NF161146)/ ; //Global Challenges Research Fund grant (BB/P023444/1)/ ; //the Agricultural Science and Technology Innovation Program (CAAS-ZDRW202007 and ASTIP-TRIC04)/ ; //the National Natural Science Foundation of China (31901893)/ ; }, mesh = {Animals ; *Baculoviridae ; Egypt ; Larva ; Pupa ; Spodoptera ; }, abstract = {BACKGROUND: The general principle of using microbes from one species to manage a different pest species has a clear precedent in the large-scale release of mosquitoes carrying a Wolbachia bacterium derived from Drosophila flies. New technologies will facilitate the discovery of microbes that can be used in a similar way. Previously, we found three novel partiti-like viruses in the African armyworm (Spodoptera exempta). To investigate further the utility and consistency of host shift of insect viruses as a potential pest management tool, we tested the interaction between the partiti-like viruses and another novel host, the Egyptian cotton leafworm (Spodoptera littoralis).

RESULT: We found that all three partiti-like viruses appeared to be harmful to the novel host S. littoralis, by causing increased larval and pupal mortality. No effect was observed on host fecundity, and partiti-like virus infection did not impact host susceptibility when challenged with another pathogen, the baculovirus SpliNPV. Transcriptome analysis of partiti-like virus-infected and noninfected S. littoralis indicated that the viruses could impact host gene-expression profiles of S. littoralis, but they impact different pathways to the two other Spodoptera species through effects on pathways related to immunity (Jak-STAT/Toll and Imd) and reproduction (insulin signaling/insect hormones).

CONCLUSION: Taken together with the previous findings in the novel host S. frugiperda, these results indicate a parasitic relationship between the partiti-like viruses and novel insect hosts, suggesting a possible use and novel pest management strategy through the artificial host shift of novel viruses. © 2021 Society of Chemical Industry.}, } @article {pmid34960705, year = {2021}, author = {Paradkar, PN and Sahasrabudhe, PR and Ghag Sawant, M and Mukherjee, S and Blasdell, KR}, title = {Towards Integrated Management of Dengue in Mumbai.}, journal = {Viruses}, volume = {13}, number = {12}, pages = {}, pmid = {34960705}, issn = {1999-4915}, mesh = {Animals ; Cities ; Community Participation ; Dengue/diagnosis/*epidemiology/*prevention & control/transmission ; Disease Notification ; *Disease Outbreaks/prevention & control ; Epidemiological Monitoring ; Humans ; India/epidemiology ; Mosquito Control ; Mosquito Vectors ; Urban Health ; }, abstract = {With increasing urbanisation, the dengue disease burden is on the rise in India, especially in large cities such as Mumbai. Current dengue surveillance in Mumbai includes municipal corporation carrying out specific activities to reduce mosquito breeding sites and the use of insecticides to suppress the adult mosquito populations. Clinical cases remain either underreported or misreported due to the restriction to government clinics, missing the large private health care sector. There is a need for an integrated approach to manage dengue outbreaks in Mumbai. There are various novel strategies available for use that can be utilised to improve disease detection, mosquito surveillance, and control of mosquito-borne diseases. These novel technologies are discussed in this manuscript. Given the complex ecosystem of mosquito-borne diseases in Mumbai, integrating data obtained from these technologies would support the ongoing mosquito control measures in Mumbai.}, } @article {pmid34954414, year = {2022}, author = {Namias, A and Sicard, M and Weill, M and Charlat, S}, title = {From Wolbachia genomics to phenotype: molecular models of cytoplasmic incompatibility must account for the multiplicity of compatibility types.}, journal = {Current opinion in insect science}, volume = {49}, number = {}, pages = {78-84}, doi = {10.1016/j.cois.2021.12.005}, pmid = {34954414}, issn = {2214-5753}, mesh = {Animals ; Antidotes ; Genomics ; Male ; Models, Molecular ; Phenotype ; *Wolbachia/genetics ; }, abstract = {Wolbachia endosymbionts commonly induce cytoplasmic incompatibility, making infected males' sperm lethal to the embryos unless these are rescued by the same bacterium, inherited from their mother. Causal genes were recently identified but two families of mechanistic models are still opposed. In the toxin-antidote model, interaction between the toxin and the antidote is required for rescuing the embryos. In host modification models, a host factor is misregulated in sperm and rescue occurs through compensation or withdrawal of this modification. While these models have been thoroughly discussed, the multiplicity of compatibility types, that is, the existence of many mutually incompatible strains, as seen in Culex mosquitoes, has not received sufficient attention. To explain such a fact, host modification models must posit that the same embryonic defects can be induced and rescued through a large variety of host targets. Conversely, the toxin-antidote model simply accommodates this pattern in a lock-key fashion, through variations in the toxin-antidote interaction sites.}, } @article {pmid34953157, year = {2022}, author = {Røed, ES and Engelstädter, J}, title = {Cytoplasmic incompatibility in hybrid zones: infection dynamics and resistance evolution.}, journal = {Journal of evolutionary biology}, volume = {35}, number = {2}, pages = {240-253}, doi = {10.1111/jeb.13974}, pmid = {34953157}, issn = {1420-9101}, mesh = {Animals ; *Arthropods ; Cytoplasm ; Female ; Male ; Reproduction ; Symbiosis ; *Wolbachia ; }, abstract = {Cytoplasmic incompatibility is an endosymbiont-induced mating incompatibility common in arthropods. Unidirectional cytoplasmic incompatibility impairs crosses between infected males and uninfected females, whereas bidirectional cytoplasmic incompatibility occurs when two host lineages are infected with reciprocally incompatible endosymbionts. Bidirectional cytoplasmic incompatibility is unstable in unstructured populations, but may be stable in hybrid zones. Stable coexistence of incompatible host lineages should generate frequent incompatible crosses. Therefore, hosts are expected to be under selection to resist their endosymbionts. Here, we formulate a mathematical model of hybrid zones where two bidirectionally incompatible host lineages meet. We expand this model to consider the invasion of a hypothetical resistance allele. To corroborate our mathematical predictions, we test each prediction with stochastic, individual-based simulations. Our models suggest that hybrid zones may sustain stable coinfections of bidirectionally incompatible endosymbiont strains. Over a range of conditions, hosts are under selection for resistance against cytoplasmic incompatibility. Under asymmetric migration, a resistance allele can facilitate infection turnover and subsequently either persist or become lost. The predictions we present may inform our understanding of the cophylogenetic relationship between the endosymbiont Wolbachia and its hosts.}, } @article {pmid34949167, year = {2021}, author = {Arham, AF and Amin, L and Mustapa, MAC and Mahadi, Z and Yaacob, M and Ibrahim, M}, title = {Determinants of stakeholders' attitudes and intentions toward supporting the use of Wolbachia-infected Aedes mosquitoes for dengue control.}, journal = {BMC public health}, volume = {21}, number = {1}, pages = {2314}, pmid = {34949167}, issn = {1471-2458}, mesh = {*Aedes ; Animals ; Attitude ; *Dengue/prevention & control ; Humans ; Intention ; *Wolbachia ; }, abstract = {BACKGROUND: A recent approach in controlling dengue is by using the Wolbachia-infected Aedes mosquito (WiAM). The approach has been reported to be more effective than traditional methods, such as fogging. Therefore, it is imperative to assess the factors predicting its acceptance among stakeholders before implementing this technology more widely in Malaysia.

METHODS: The survey data were collected from two primary stakeholder groups using a stratified random sampling technique. The two primary stakeholder groups were scientists (n = 202) and the public (n = 197) in the Klang Valley region of Malaysia, a hot spot area known for the high rate of dengue cases. The respondents answered questions on a seven-point Likert scale survey regarding trust in key players, attitudes toward nature versus materialism, religiosity, perceived benefits, perceived risks, attitudes, and intentions. The data were analyzed using Smart Partial Least Square (SmartPLS) software (version 3.2.6) to determine the predictors influencing attitudes and intentions to support the use of WiAM technology.

RESULTS: The results indicated a strong positive relationship between attitudes and intentions to support the use of WiAM (β = 0.676, p < 0.001). The most important significant predictor for attitudes was perceived benefits (β = 0.493, p < 0.001), followed by perceived risks (β = - 0.080, p = 0.048). Trust in key players, attitudes toward nature versus material, and religiosity had indirect relationships with attitudes through the perceived benefits and risks.

CONCLUSIONS: The identified predictors can serve as indicators for the decision-making process regarding WiAM implementation in Malaysia and other developing countries with similar demographics and cultures.}, } @article {pmid34940221, year = {2021}, author = {Bamou, R and Diarra, AZ and Mayi, MPA and Djiappi-Tchamen, B and Antonio-Nkondjio, C and Parola, P}, title = {Wolbachia Detection in Field-Collected Mosquitoes from Cameroon.}, journal = {Insects}, volume = {12}, number = {12}, pages = {}, pmid = {34940221}, issn = {2075-4450}, support = {ANR-10-IAHU-03//National Research Agency under the program « Investissements d'avenir »/ ; FEDER PRIMI//Région Provence Alpes Côte d'Azur and European funding/ ; }, abstract = {Wolbachia spp., known to be maternally inherited intracellular bacteria, are widespread among arthropods, including mosquitoes. Our study assessed the presence and prevalence of Wolbachia infection in wild mosquitoes collected in Cameroon, using the combination of 23s rRNA Anaplasmatacea and 16s rRNA Wolbachia genes. Mosquitoes that were positive for Wolbachia were sequenced for subsequent phylogenetic analysis. Out of a total of 1740 individual mosquitoes belonging to 22 species and five genera screened, 33 mosquitoes (1.87%) belonging to eight species (namely, Aedes albopictus, A. contigus, Culex quinquefasciatus, C. perfuscus, C. wigglesworthi, C. duttoni, Anopheles paludis and Coquillettidia sp.) were found to be positive for Wolbachia infections. Wolbachia spp. were absent in A. gambiae and A. aegypti, the main vectors of malaria and dengue, respectively. Phylogenetic analysis of the 16S RNA sequences showed they belong mainly to two distinct subgroups (A and B). This study reports the presence of Wolbachia in about eight species of mosquitoes in Cameroon and suggests that future characterisation of the strains is needed.}, } @article {pmid34940211, year = {2021}, author = {Beribaka, M and Jelić, M and Tanasković, M and Lazić, C and Stamenković-Radak, M}, title = {Life History Traits in Two Drosophila Species Differently Affected by Microbiota Diversity under Lead Exposure.}, journal = {Insects}, volume = {12}, number = {12}, pages = {}, pmid = {34940211}, issn = {2075-4450}, support = {19/6-020/961-104/18//Ministry for Scientific and Technological Development, Higher Education and Information Society of the Republic of Srpska, B&H/ ; 451-03-9/2021-14/200178//Ministry of Education, Science and Technological Development of the Republic of Serbia/ ; 451-03-9/2021-14/ 200007//Ministry of Education, Science and Technological Development of the Republic of Serbia/ ; }, abstract = {Life history traits determine the persistence and reproduction of each species. Factors that can affect life history traits are numerous and can be of different origin. We investigated the influence of population origin and heavy metal exposure on microbiota diversity and two life history traits, egg-to-adult viability and developmental time, in Drosophila melanogaster and Drosophila subobscura, grown in the laboratory on a lead (II) acetate-saturated substrate. We used 24 samples, 8 larval and 16 adult samples (two species × two substrates × two populations × two sexes). The composition of microbiota was determined by sequencing (NGS) of the V3-V4 variable regions of the 16S rRNA gene. The population origin showed a significant influence on life history traits, though each trait in the two species was affected differentially. Reduced viability in D. melanogaster could be a cost of fast development, decrease in Lactobacillus abundance and the presence of Wolbachia. The heavy metal exposure in D. subobscura caused shifts in developmental time but maintained the egg-to-adult viability at a similar level. Microbiota diversity indicated that the Komagataeibacter could be a valuable member of D. subobscura microbiota in overcoming the environmental stress. Research on the impact of microbiota on the adaptive response to heavy metals and consequently the potential tradeoffs among different life history traits is of great importance in evolutionary research.}, } @article {pmid34940142, year = {2021}, author = {Jiang, RX and Shang, F and Jiang, HB and Dou, W and Cernava, T and Wang, JJ}, title = {The Influence of Temperature and Host Gender on Bacterial Communities in the Asian Citrus Psyllid.}, journal = {Insects}, volume = {12}, number = {12}, pages = {}, pmid = {34940142}, issn = {2075-4450}, abstract = {The Asian citrus psyllid, D. citri Kuwayama is the primary vector for Candidatus Liberibacter asiaticus (CLas), which causes a destructive disease in citrus plants. Bacterial symbionts are important determinants of insect physiology, and they can be impacted by many external factors. Temperature is an important abiotic factor affecting insect physiology, and it is also known that differences in symbiont proportions may vary in different insect genders. To date, it is unclear how the symbionts of D. citri are affected by temperature and gender. This study used high-throughput sequencing of 16S ribosomal RNA amplicons to determine how temperature and gender affect the bacterial communities present in D. citri. We identified 27 amplicon sequence variants (ASVs) belonging to 10 orders, seven classes, and five phyla. The dominant phylum was Proteobacteria (99.93%). Other phyla, including Firmicutes, Bacteroidota, Deinococcota, Cyanobacteria, and Actinobacteriota, were less abundant (<0.1%). Profftella (71.77-81.59%) and Wolbachia (18.39-28.22%) were the predominant taxa in all samples. Under high-temperature treatment, Profftella was more common in females, while Wolbachia had a higher abundance in males. In males, Profftella was more abundant under low-temperature treatments than under high-temperature treatments. In contrast, Wolbachia showed a higher abundance under high-temperature treatments than under low-temperature treatments. An RT-qPCR (quantitative real-time PCR) approach confirmed the results obtained with high-throughput DNA sequencing. Our results provide a basis for understanding the co-adaptation of D. citri and its symbionts to environmental temperature stress.}, } @article {pmid34933167, year = {2022}, author = {Aouadi, N and Benkacimi, L and Zan Diarra, A and Laroche, M and Bérenger, JM and Bitam, I and Parola, P}, title = {Microorganisms associated with the North African hedgehog Atelerix algirus and its parasitizing arthropods in Algeria.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {80}, number = {}, pages = {101726}, doi = {10.1016/j.cimid.2021.101726}, pmid = {34933167}, issn = {1878-1667}, mesh = {Algeria/epidemiology ; Animals ; *Arthropods ; Hedgehogs ; *Rhipicephalus sanguineus ; *Siphonaptera ; }, abstract = {Hedgehogs are small mammals. They are potential reservoirs of various zoonotic agents. This study was conducted in Bouira, a north-central region of Algeria. A total of 21 Atelerix algirus corpses were picked up on roadsides and gardens. Hedgehog kidneys, spleens and ectoparasites were collected. Twelve hedgehogs were infested with ectoparasites, including Archaeopsylla erinacei, Rhipicephalus sanguineus s.l. and Haemaphysalis erinacei. Hedgehog organs and randomly selected arthropods were screened for microorganisms using molecular methods. Coxiella burnetii was detected in kidneys, spleens, A. erinacei, Hae. erinacei and Rh. sanguineus s.l. Leptospira interrogans was detected in kidneys. Rickettsia felis and Rickettsia massiliae were detected respectively in A. erinacei and in Rh. sanguineus s.l. DNA of an uncultivated Rickettsia spp. was found in Hae. erinacei. Wolbachia spp. DNA was detected in fleas. The DNA of potential new Bartonella and Ehrlichia species were found respectively in fleas and ticks. This study highlights the presence of DNA from a broad range of microorganisms in hedgehogs and their ectoparasites that may be responsible for zoonoses in Algeria.}, } @article {pmid34928947, year = {2021}, author = {Vivero-Gomez, RJ and Castañeda-Monsalve, VA and Atencia, MC and Hoyos-Lopez, R and Hurst, GD and Cadavid-Restrepo, G and Moreno-Herrera, CX}, title = {Molecular phylogeny of heritable symbionts and microbiota diversity analysis in phlebotominae sand flies and Culex nigripalpus from Colombia.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {12}, pages = {e0009942}, pmid = {34928947}, issn = {1935-2735}, mesh = {Animals ; Bacteroidetes/classification/genetics/*isolation & purification/physiology ; Biodiversity ; Colombia ; Culex/*microbiology/physiology ; *Microbiota ; Microsporidia/classification/genetics/*isolation & purification/physiology ; *Phylogeny ; Psychodidae/*microbiology/physiology ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {BACKGROUND: Secondary symbionts of insects include a range of bacteria and fungi that perform various functional roles on their hosts, such as fitness, tolerance to heat stress, susceptibility to insecticides and effects on reproduction. These endosymbionts could have the potential to shape microbial communites and high potential to develop strategies for mosquito-borne disease control.

The relative frequency and molecular phylogeny of Wolbachia, Microsporidia and Cardinium were determined of phlebotomine sand flies and mosquitoes in two regions from Colombia. Illumina Miseq using the 16S rRNA gene as a biomarker was conducted to examine the microbiota. Different percentages of natural infection by Wolbachia, Cardinium, and Microsporidia in phlebotomines and mosquitoes were detected. Phylogenetic analysis of Wolbachia shows putative new strains of Lutzomyia gomezi (wLgom), Brumptomyia hamata (wBrham), and a putative new group associated with Culex nigripalpus (Cnig) from the Andean region, located in Supergroup A and Supergroup B, respectively. The sequences of Microsporidia were obtained of Pi. pia and Cx. nigripalpus, which are located on phylogeny in the IV clade (terrestrial origin). The Cardinium of Tr. triramula and Ps. shannoni were located in group C next to Culicoides sequences while Cardinium of Mi. cayennensis formed two putative new subgroups of Cardinium in group A. In total were obtained 550 bacterial amplicon sequence variants (ASVs) and 189 taxa to the genus level. The microbiota profiles of Sand flies and mosquitoes showed mainly at the phylum level to Proteobacteria (67.6%), Firmicutes (17.9%) and Actinobacteria (7.4%). High percentages of relative abundance for Wolbachia (30%-83%) in Lu. gomezi, Ev. dubitans, Mi. micropyga, Br. hamata, and Cx. nigripalpus were found. ASVs assigned as Microsporidia were found in greater abundance in Pi. pia (23%) and Cx. nigripalpus (11%). An important finding is the detection of Rickettsia in Pi. pia (58,8%) and Bartonella sp. in Cx. nigripalpus.

CONCLUSIONS/SIGNIFICANCE: We found that Wolbachia infection significantly decreased the alpha diversity and negatively impacts the number of taxa on sand flies and Culex nigripalpus. The Principal Coordinate Analysis (PCoA) is consistent, which showed statistically significant differences (PERMANOVA, F = 2.4744; R2 = 0.18363; p-value = 0.007) between the microbiota of sand flies and mosquitoes depending on its origin, host and possibly for the abundance of some endosymbionts (Wolbachia, Rickettsia).}, } @article {pmid34919808, year = {2022}, author = {Hague, MTJ and Shropshire, JD and Caldwell, CN and Statz, JP and Stanek, KA and Conner, WR and Cooper, BS}, title = {Temperature effects on cellular host-microbe interactions explain continent-wide endosymbiont prevalence.}, journal = {Current biology : CB}, volume = {32}, number = {4}, pages = {878-888.e8}, pmid = {34919808}, issn = {1879-0445}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; Host Microbial Interactions ; Prevalence ; Temperature ; *Wolbachia/genetics ; }, abstract = {Endosymbioses influence host physiology, reproduction, and fitness, but these relationships require efficient microbe transmission between host generations to persist. Maternally transmitted Wolbachia are the most common known endosymbionts,[1] but their frequencies vary widely within and among host populations for unknown reasons.[2][,][3] Here, we integrate genomic, cellular, and phenotypic analyses with mathematical models to provide an unexpectedly simple explanation for global wMel Wolbachia prevalence in Drosophila melanogaster. Cooling temperatures decrease wMel cellular abundance at a key stage of host oogenesis, producing temperature-dependent variation in maternal transmission that plausibly explains latitudinal clines of wMel frequencies on multiple continents. wMel sampled from a temperate climate targets the germline more efficiently in the cold than a recently differentiated tropical variant (∼2,200 years ago), indicative of rapid wMel adaptation to climate. Genomic analyses identify a very narrow list of wMel alleles-most notably, a derived stop codon in the major Wolbachia surface protein WspB-that underlie thermal sensitivity of cellular Wolbachia abundance and covary with temperature globally. Decoupling temperate wMel and host genomes further reduces transmission in the cold, a pattern that is characteristic of host-microbe co-adaptation to a temperate climate. Complex interactions among Wolbachia, hosts, and the environment (GxGxE) mediate wMel cellular abundance and maternal transmission, implicating temperature as a key determinant of Wolbachia spread and equilibrium frequencies, in conjunction with Wolbachia effects on host fitness and reproduction.[4][,][5] Our results motivate the strategic use of locally selected wMel variants for Wolbachia-based biocontrol efforts, which protect millions of individuals from arboviruses that cause human disease.[6].}, } @article {pmid34906084, year = {2021}, author = {Lau, MJ and Schmidt, TL and Yang, Q and Chung, J and Sankey, L and Ross, PA and Hoffmann, AA}, title = {Genetic stability of Aedes aegypti populations following invasion by wMel Wolbachia.}, journal = {BMC genomics}, volume = {22}, number = {1}, pages = {894}, pmid = {34906084}, issn = {1471-2164}, mesh = {*Aedes/genetics ; Animals ; *Arboviruses ; *Wolbachia/genetics ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {BACKGROUND: Wolbachia wMel is the most commonly used strain in rear and release strategies for Aedes aegypti mosquitoes that aim to inhibit the transmission of arboviruses such as dengue, Zika, Chikungunya and yellow fever. However, the long-term establishment of wMel in natural Ae. aegypti populations raises concerns that interactions between Wolbachia wMel and Ae. aegypti may lead to changes in the host genome, which could affect useful attributes of Wolbachia that allow it to invade and suppress disease transmission.

RESULTS: We applied an evolve-and-resequence approach to study genome-wide genetic changes in Ae. aegypti from the Cairns region, Australia, where Wolbachia wMel was first introduced more than 10 years ago. Mosquito samples were collected at three different time points in Gordonvale, Australia, covering the phase before (2010) and after (2013 and 2018) Wolbachia releases. An additional three locations where Wolbachia replacement happened at different times across the last decade were also sampled in 2018. We found that the genomes of mosquito populations mostly remained stable after Wolbachia release, with population differences tending to reflect the geographic location of the populations rather than Wolbachia infection status. However, outlier analysis suggests that Wolbachia may have had an influence on some genes related to immune response, development, recognition and behavior.

CONCLUSIONS: Ae. aegypti populations remained geographically distinct after Wolbachia wMel releases in North Australia despite their Wolbachia infection status. At some specific genomic loci, we found signs of selection associated with Wolbachia, suggesting potential evolutionary impacts can happen in the future and further monitoring is warranted.}, } @article {pmid34903056, year = {2021}, author = {Shropshire, JD and Hamant, E and Cooper, BS}, title = {Male Age and Wolbachia Dynamics: Investigating How Fast and Why Bacterial Densities and Cytoplasmic Incompatibility Strengths Vary.}, journal = {mBio}, volume = {12}, number = {6}, pages = {e0299821}, pmid = {34903056}, issn = {2150-7511}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/genetics/*microbiology ; Drosophila/genetics/immunology/*microbiology/physiology ; Drosophila melanogaster ; Female ; Male ; Species Specificity ; Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Endosymbionts can influence host reproduction and fitness to favor their maternal transmission. For example, endosymbiotic Wolbachia bacteria often cause cytoplasmic incompatibility (CI) that kills uninfected embryos fertilized by Wolbachia-modified sperm. Infected females can rescue CI, providing them a relative fitness advantage. Wolbachia-induced CI strength varies widely and tends to decrease as host males age. Since strong CI drives Wolbachia to high equilibrium frequencies, understanding how fast and why CI strength declines with male age is crucial to explaining age-dependent CI's influence on Wolbachia prevalence. Here, we investigate if Wolbachia densities and/or CI gene (cif) expression covary with CI-strength variation and explore covariates of age-dependent Wolbachia-density variation in two classic CI systems. wRi CI strength decreases slowly with Drosophila simulans male age (6%/day), but wMel CI strength decreases very rapidly (19%/day), yielding statistically insignificant CI after only 3 days of Drosophila melanogaster adult emergence. Wolbachia densities and cif expression in testes decrease as wRi-infected males age, but both surprisingly increase as wMel-infected males age, and CI strength declines. We then tested if phage lysis, Octomom copy number (which impacts wMel density), or host immune expression covary with age-dependent wMel densities. Only host immune expression correlated with density. Together, our results identify how fast CI strength declines with male age in two model systems and reveal unique relationships between male age, Wolbachia densities, cif expression, and host immunity. We discuss new hypotheses about the basis of age-dependent CI strength and its contributions to Wolbachia prevalence. IMPORTANCEWolbachia bacteria are the most common animal-associated endosymbionts due in large part to their manipulation of host reproduction. Many Wolbachia cause cytoplasmic incompatibility (CI) that kills uninfected host eggs. Infected eggs are protected from CI, favoring Wolbachia spread in natural systems and in transinfected mosquito populations where vector-control groups use strong CI to maintain pathogen-blocking Wolbachia at high frequencies for biocontrol of arboviruses. CI strength varies considerably in nature and declines as males age for unknown reasons. Here, we determine that CI strength weakens at different rates with age in two model symbioses. Wolbachia density and CI gene expression covary with wRi-induced CI strength in Drosophila simulans, but neither explain rapidly declining wMel-induced CI in aging D. melanogaster males. Patterns of host immune gene expression suggest a candidate mechanism behind age-dependent wMel densities. These findings inform how age-dependent CI may contribute to Wolbachia prevalence in natural systems and potentially in transinfected systems.}, } @article {pmid34871301, year = {2021}, author = {Sánchez-González, L and Adams, LE and Saavedra, R and Little, EM and Medina, NA and Major, CG and Bruck, M and Miranda, J and Rosado-Santiago, C and Ryff, K and Ortiz, M and Brown, G and Barrera, R and Pérez-Guerra, CL and Rivera-Amill, V and Paz-Bailey, G}, title = {Assessment of community support for Wolbachia-mediated population suppression as a control method for Aedes aegypti mosquitoes in a community cohort in Puerto Rico.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {12}, pages = {e0009966}, pmid = {34871301}, issn = {1935-2735}, support = {U01 CK000437/CK/NCEZID CDC HHS/United States ; U01 CK000580/CK/NCEZID CDC HHS/United States ; U01CK000580/ACL/ACL HHS/United States ; }, mesh = {Adult ; Aedes/*microbiology/physiology ; Animals ; Cohort Studies ; Community Support/psychology ; Female ; Humans ; Male ; Middle Aged ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology/physiology ; Sexual Behavior, Animal ; Surveys and Questionnaires ; Wolbachia/*physiology ; Young Adult ; }, abstract = {Arboviral diseases transmitted by Aedes species mosquitoes pose an increasing public health challenge in tropical regions. Wolbachia-mediated population suppression (Wolbachia suppression) is a vector control method used to reduce Aedes mosquito populations by introducing male mosquitoes infected with Wolbachia, a naturally occurring endosymbiotic bacterium. When Wolbachia-infected male mosquitoes mate with female wild mosquitoes, the resulting eggs will not hatch. Public support is vital to the successful implementation and sustainability of vector control interventions. Communities Organized to Prevent Arboviruses (COPA) is a cohort study to determine the incidence of arboviral disease in Ponce, Puerto Rico and evaluate vector control methods. Focus groups were conducted with residents of COPA communities to gather their opinion on vector control methods; during 2018-2019, adult COPA participants were interviewed regarding their views on Wolbachia suppression; and a follow-up questionnaire was conducted among a subset of participants and non-participants residing in COPA communities. We analyzed factors associated with support for this method. Among 1,528 participants in the baseline survey, median age was 37 years and 63% were female. A total of 1,032 (68%) respondents supported Wolbachia suppression. Respondents with an income of $40,000 or more were 1.34 times as likely [95% CI: 1.03, 1.37] to support Wolbachia suppression than those who earned less than $40,000 annually. Respondents who reported repellant use were 1.19 times as likely to support Wolbachia suppression [95% CI: 1.03, 1.37]. A follow-up survey in 2020 showed that most COPA participants (86%) and non-participants living in COPA communities (84%) supported Wolbachia suppression during and after an educational campaign. The most frequent questions regarding this method were related to its impact on human and animal health, and the environment. Continuous community engagement and education efforts before and during the implementation of novel vector control interventions are necessary to increase and maintain community support.}, } @article {pmid34865032, year = {2021}, author = {Wangkeeree, J and Sanit, P and Roddee, J and Hanboonsong, Y}, title = {Population Dynamics of Wolbachia in the Leafhopper Vector Yamatotettix flavovittatus (Hemiptera: Cicadellidae).}, journal = {Journal of insect science (Online)}, volume = {21}, number = {6}, pages = {}, pmid = {34865032}, issn = {1536-2442}, support = {226/2563//National Research Council of Thailand/ ; }, mesh = {Animals ; *Hemiptera/microbiology ; Population Dynamics ; Saccharum ; *Wolbachia ; }, abstract = {Wolbachia (Rickettsiales: Alphaproteobacteria) infections induce abnormalities in the reproductive system and affect various biological traits of the host insects. The density of Wolbachia is one of the major parameters that influence induced phenotypes and interactions with the hosts. Wolbachia occurs naturally in populations of the leafhopper Yamatotettix flavovittatus Matsumura (Hemiptera: Cicadellidae), which transmits phytoplasma that cause white leaf disease in sugarcane. However, the quantity and dynamics of Wolbachia in this leafhopper are not well understood. In the current study, we estimated the number of Wolbachia by absolute quantification of the copy number of wsp, which encodes the outer surface protein, using real-time quantitative polymerase chain reaction (PCR). This investigation was performed using natural populations and laboratory colonies from three lineages of leafhoppers (designated as UD, KK, and SK). There was no significant difference in the number of wsp copies in most of field-collected adults. During the immature developmental stages, there were differences in the dynamics of Wolbachia infection between the UD lineage and the other two lineages. However, the number of wsp copies increased in the early instar and plateaued in the later nymphal instars. Sex had no influence on the number of Wolbachia within the same lineages. The number of Wolbachia was relatively constant during the adult stage in the UD lineage but fluctuated in the other two lineages. In conclusion, the present data provide a framework for exploring the relationship between Wolbachia and the leafhopper and could facilitate future research into management strategies using Wolbachia.}, } @article {pmid34864906, year = {2022}, author = {Cooper, WR and Horton, DR and Swisher-Grimm, K and Krey, K and Wildung, MR}, title = {Bacterial Endosymbionts of Bactericera maculipennis and Three Mitochondrial Haplotypes of B. cockerelli (Hemiptera: Psylloidea: Triozidae).}, journal = {Environmental entomology}, volume = {51}, number = {1}, pages = {94-107}, doi = {10.1093/ee/nvab133}, pmid = {34864906}, issn = {1938-2936}, mesh = {Animals ; Bacteria/genetics ; Haplotypes ; *Hemiptera/microbiology ; Plant Diseases/microbiology ; *Rhizobiaceae ; *Solanum tuberosum/microbiology ; }, abstract = {Insects harbor bacterial endosymbionts that provide their hosts with nutritional benefit or with protection against natural enemies, plant defenses, insecticides, or abiotic stresses. We used directed sequencing of 16S rDNA to identify and compare endosymbionts of Bactericera maculipennis (Crawford) and the western, central, and northwestern haplotypes of B. cockerelli (Šulc) (Hemiptera: Psylloidea: Triozidae). Both species are native to North America, are known to harbor the plant pathogen 'Candidatus Liberibacter solanacearum' and develop on shared host plants within the Convolvulaceae. The Old-World species Heterotrioza chenopodii (Reuter) (Psylloidea: Triozidae), now found in North America, was included as an outgroup. 16S sequencing confirmed that both Bactericera species harbor 'Candidatus Liberibacter solanacearum' and revealed that both species harbor unique strains of Wolbachia and Sodalis. However, the presence of Wolbachia and Sodalis varied among haplotypes of B. cockerelli. The central and western haplotypes harbored the same strains of Wolbachia, which was confirmed by Sanger sequencing of the wsp and ftsZ genes. Wolbachia was also detected in very low abundance from the northwestern haplotype by high-throughput sequencing of 16S but was not detected from this haplotype by PCR screening. The northwestern and central haplotypes also harbored Sodalis, which was not detected in the western haplotype. Heterotrioza chenopodii harbored an entirely different community of potential endosymbionts compared with the Bactericera spp. that included Rickettsia and an unidentified bacterium in the Enterobacteriaceae. Results of this study provide a foundation for further research on the interactions between psyllids and their bacterial endosymbionts.}, } @article {pmid34862932, year = {2021}, author = {Girardin, L and Débarre, F}, title = {Demographic feedbacks can hamper the spatial spread of a gene drive.}, journal = {Journal of mathematical biology}, volume = {83}, number = {6-7}, pages = {67}, pmid = {34862932}, issn = {1432-1416}, mesh = {Feedback ; *Gene Drive Technology ; Humans ; Population Dynamics ; }, abstract = {This paper is concerned with a reaction-diffusion system modeling the fixation and the invasion in a population of a gene drive (an allele biasing inheritance, increasing its own transmission to offspring). In our model, the gene drive has a negative effect on the fitness of individuals carrying it, and is therefore susceptible of decreasing the total carrying capacity of the population locally in space. This tends to generate an opposing demographic advection that the gene drive has to overcome in order to invade. While previous reaction-diffusion models neglected this aspect, here we focus on it and try to predict the sign of the traveling wave speed. It turns out to be an analytical challenge, only partial results being within reach, and we complete our theoretical analysis by numerical simulations. Our results indicate that taking into account the interplay between population dynamics and population genetics might actually be crucial, as it can effectively reverse the direction of the invasion and lead to failure. Our findings can be extended to other bistable systems, such as the spread of cytoplasmic incompatibilities caused by Wolbachia.}, } @article {pmid34860833, year = {2021}, author = {Gil, MF and Fassolari, M and Battaglia, ME and Berón, CM}, title = {Culex quinquefasciatus larvae development arrested when fed on Neochloris aquatica.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {12}, pages = {e0009988}, pmid = {34860833}, issn = {1935-2735}, mesh = {Animals ; Culex/*growth & development/*microbiology/physiology ; Feeding Behavior ; Female ; Larva/*microbiology ; *Microalgae ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology/physiology ; Oviposition ; Wolbachia/isolation & purification ; }, abstract = {Culex quinquefasciatus is a cosmopolitan species widely distributed in the tropical and subtropical areas of the world. Due to its long history of close association with humans, the transmission of arboviruses and parasites have an important role in veterinary and public health. Adult females feed mainly on birds although they can also feed on humans and other mammals. On the other hand, larvae are able to feed on a great diversity of microorganisms, including microalgae, present in natural or artificial breeding sites with a high organic load. These two particularities, mentioned above, are some of the reasons why this mosquito is so successful in the environment. In this work, we report the identification of a microalga found during field sampling in artificial breeding sites, in a group of discarded tires with accumulated rainwater. Surprisingly, only one of them had a bright green culture without mosquito larvae while the other surrounding tires contained a large number of mosquito larvae. We isolated and identified this microorganism as Neochloris aquatica, and it was evaluated as a potential biological control agent against Cx. quinquefasciatus. The oviposition site preference in the presence of the alga by gravid females, and the effects on larval development were analyzed. Additionally, microalga effect on Cx. quinquefasciatus wild type, naturally infected with the endosymbiotic bacterium Wolbachia (w+) and Wolbachia free (w-) laboratory lines was explored. According to our results, even though it is chosen by gravid females to lay their eggs, the microalga had a negative effect on the development of larvae from both populations. Additionally, when the larvae were fed with a culture of alga supplemented with balanced fish food used as control diet, they were not able to reverse its effect, and were unable to complete development until adulthood. Here, N. aquatica is described as a biological agent, and as a potential source of bioactive compounds for the control of mosquito populations important in veterinary and human health.}, } @article {pmid34852835, year = {2021}, author = {Díaz, S and Camargo, C and Avila, FW}, title = {Characterization of the reproductive tract bacterial microbiota of virgin, mated, and blood-fed Aedes aegypti and Aedes albopictus females.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {592}, pmid = {34852835}, issn = {1756-3305}, support = {566-1-2014//Departamento Administrativo de Ciencia, Tecnología e Innovación/ ; }, mesh = {Aedes/*microbiology ; Animals ; Bacteria/*classification/genetics/isolation & purification ; DNA Barcoding, Taxonomic ; Female ; Genitalia/microbiology ; Humans ; *Microbiota ; Mosquito Vectors/*parasitology ; Organ Specificity ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; }, abstract = {BACKGROUND: Aedes aegypti and Ae. albopictus are vectors of numerous arboviruses that adversely affect human health. In mosquito vectors of disease, the bacterial microbiota influence several physiological processes, including fertility and vector competence, making manipulation of the bacterial community a promising method to control mosquito vectors. In this study, we describe the reproductive tract tissue microbiota of lab-reared virgin Ae. aegypti and Ae. albopictus males, and virgin, mated, and mated + blood-fed females of each species, comparing the bacterial composition found there to the well-described gut microbiota.

METHODS: We performed metabarcoding of the 16S rRNA isolated from the gut, upper reproductive tract (URT; testes or ovaries), and lower reproductive tract (LRT; males: seminal vesicles and accessory glands; females: oviduct, spermathecae, and bursa) for each species, and evaluated the influence of host species, tissue, nutritional status, and reproductive status on microbiota composition. Finally, based on the identified taxonomic profiles of the tissues assessed, bacterial metabolic pathway abundance was predicted.

RESULTS: The community structure of the reproductive tract is unique compared to the gut. Asaia is the most prevalent OTU in the LRTs of both Ae. aegypti and Ae. albopictus. In the URT, we observed differences between species, with Wolbachia OTUs being dominant in the Ae. albopictus URT, while Enterobacter and Serratia were dominant in Ae. aegypti URT. Host species and tissue were the best predictors of the community composition compared to reproductive status (i.e., virgin or mated) and nutritional status (i.e., sugar or blood-fed). The predicted functional profile shows changes in the abundance of specific microbial pathways that are associated with mating and blood-feeding, like energy production in mated tissues and siderophore synthesis in blood-fed female tissues.

CONCLUSIONS: Aedes aegypti and Ae. albopictus have distinct differences in the composition of microbiota found in the reproductive tract. The distribution of the bacterial taxonomic groups indicates that some bacteria have tissue-specific tropism for reproductive tract tissue, such as Asaia and Wolbachia. No significant differences in the taxonomic composition were observed in the reproductive tract between virgin, mated, and mated + blood-fed females, but changes in the abundance of specific metabolic pathways were found in the predicted microbial functional profiles in mated and blood-fed females.}, } @article {pmid34850518, year = {2022}, author = {Brunetti, M and Magoga, G and Gionechetti, F and De Biase, A and Montagna, M}, title = {Does diet breadth affect the complexity of the phytophagous insect microbiota? The case study of Chrysomelidae.}, journal = {Environmental microbiology}, volume = {24}, number = {8}, pages = {3565-3579}, pmid = {34850518}, issn = {1462-2920}, mesh = {Animals ; *Aphids/microbiology ; *Coleoptera/microbiology ; Diet ; *Microbiota/genetics ; Plants/genetics ; RNA, Ribosomal, 16S/genetics ; *Rickettsia/genetics ; Symbiosis ; }, abstract = {Chrysomelidae is a family of phytophagous insects with a highly variable degree of trophic specialization. The aim of this study is to test whether species feeding on different plants (generalists) harbour more complex microbiotas than those feeding on a few or a single plant species (specialists). The microbiota of representative leaf beetle species was characterized with a metabarcoding approach targeting V1-V2 and V4 regions of the bacterial 16S rRNA. Almost all the analysed species harbour at least one reproductive manipulator bacteria (e.g., Wolbachia, Rickettsia). Two putative primary symbionts, previously isolated only from a single species (Bromius obscurus), have been detected in two species of the same subfamily, suggesting a widespread symbiosis in Eumolpinae. Surprisingly, the well-known aphid symbiont Buchnera is well represented in the microbiota of Orsodacne humeralis. Moreover, in this study, using Hill numbers to dissect the components of the microbiota diversity (abundant and rare bacteria), it has been demonstrated that generalist insect species harbour a more diversified microbiota than specialists. The higher microbiota diversity associated with a wider host-plant spectrum could be seen as an adaptive trait, conferring new metabolic potential useful to expand the diet breath, or as a result of environmental stochastic acquisition conveyed by diet.}, } @article {pmid34849818, year = {2021}, author = {Bauer DuMont, VL and White, SL and Zinshteyn, D and Aquadro, CF}, title = {Molecular population genetics of Sex-lethal (Sxl) in the Drosophila melanogaster species group: a locus that genetically interacts with Wolbachia pipientis in Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {11}, number = {8}, pages = {}, pmid = {34849818}, issn = {2160-1836}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster/genetics/metabolism ; Female ; Genetics, Population ; RNA-Binding Proteins ; *Wolbachia/metabolism ; }, abstract = {Sex-lethal (Sxl) is the sex determination switch in Drosophila, and also plays a critical role in germ-line stem cell daughter differentiation in Drosophila melanogaster. Three female-sterile alleles at Sxl in D. melanogaster were previously shown to genetically interact to varying degrees with the maternally inherited endosymbiont Wolbachia pipientis. Given this genetic interaction and W. pipientis' ability to manipulate reproduction in Drosophila, we carried out a careful study of both the population genetics (within four Drosophila species) and molecular evolutionary analysis (across 20 Drosophila species) of Sxl. Consistent with earlier studies, we find that selective constraint has played a prominent role in Sxl's molecular evolution within Drosophila, but we also observe patterns that suggest both episodic bursts of protein evolution and recent positive selection at Sxl. The episodic nature of Sxl's protein evolution is discussed in light of its genetic interaction with W. pipientis.}, } @article {pmid34843992, year = {2022}, author = {Manoj, RRS and Latrofa, MS and Bezerra-Santos, MA and Sgroi, G and Samarelli, R and Mendoza-Roldan, JA and Otranto, D}, title = {Molecular detection and characterization of the endosymbiont Wolbachia in the European hedgehog flea, Archaeopsylla erinacei.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {97}, number = {}, pages = {105161}, doi = {10.1016/j.meegid.2021.105161}, pmid = {34843992}, issn = {1567-7257}, mesh = {Animals ; Bacterial Outer Membrane Proteins/analysis ; Female ; Italy ; Male ; Phylogeny ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; Siphonaptera/*microbiology ; *Symbiosis ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia, the endosymbiont of arthropods and onchocercid nematodes is present in many medically important insect species, being also considered for the indirect control of parasitic ones. Archaeopsylla erinacei is a flea species infesting hedgehogs acting as vector of Rickettsia felis, Bartonella henselae, and Rickettsia helvetica, thus having public health relevance. The Wolbachia surface protein (wsp) and 16S rRNA genes were used to determine the presence, prevalence and molecular typing of Wolbachia in this flea species collected in two regions of southern Italy. Of the 45 fleas tested (n = 16 males, 35.6%; n = 29 females, 64.4%), 43 (95.6%; 95% CI: 84.8-99.2) scored positive for Wolbachia, of which 15 (33.3%) and 28 (62.2%) were males and females, respectively. The sex-wise prevalence of this endosymbiont was almost equal in both sexes (males 93.8%; 95% CI: 69.5-99.7; females 96.7%; 95% CI: 83.1-99.8). Single locus sequence analysis (SLST) of Wolbachia revealed two sequence types for 16S rRNA gene, named as wAr_15227 and wAr_15234, which came from two different areas, equally distributed in male and female fleas, whilst only one sequence type was identified for wsp gene. The phylogenetic analysis placed the two 16S rRNA sequence types in paraphyletic clades belonging to the supergroup A and B, respectively. Whilst, the tree of wsp gene clustered the corresponding sequence in the same clade including those of Wolbachia supergroup A. In MLST analyses, both Wolbachia sequence types clustered in a monophyletic clade with Drosophila nikananu (wNik) and Drosophila sturtevanti (wStv) from supergroup A. ClonalFrame analysis revealed a recombination event in the wAr_15234 strain which came from Apulia region. Scientific knowledge of the presence/prevalence of Wolbachia among medically important fleas, may contribute to develop an alternative biological method for the vector control.}, } @article {pmid34843477, year = {2021}, author = {Ramirez, JL and Schumacher, MK and Ower, G and Palmquist, DE and Juliano, SA}, title = {Impacts of fungal entomopathogens on survival and immune responses of Aedes albopictus and Culex pipiens mosquitoes in the context of native Wolbachia infections.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {11}, pages = {e0009984}, pmid = {34843477}, issn = {1935-2735}, support = {R15 AI124005/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/drug effects/genetics/*immunology/microbiology ; Animals ; Culex/drug effects/genetics/*immunology/microbiology ; Fungi ; Gene Expression ; Immunity/*genetics ; Insecticide Resistance ; Insecticides ; Monophenol Monooxygenase/genetics/metabolism ; Mosquito Vectors/*immunology/microbiology ; Symbiosis ; Vector Borne Diseases ; Wolbachia/*genetics ; }, abstract = {Microbial control of mosquitoes via the use of symbiotic or pathogenic microbes, such as Wolbachia and entomopathogenic fungi, are promising alternatives to synthetic insecticides to tackle the rapid increase in insecticide resistance and vector-borne disease outbreaks. This study evaluated the susceptibility and host responses of two important mosquito vectors, Ae. albopictus and Cx. pipiens, that naturally carry Wolbachia, to infections by entomopathogenic fungi. Our study indicated that while Wolbachia presence did not provide a protective advantage against entomopathogenic fungal infection, it nevertheless influenced the bacterial / fungal load and the expression of select anti-microbial effectors and phenoloxidase cascade genes in mosquitoes. Furthermore, although host responses from Ae. albopictus and Cx. pipiens were mostly similar, we observed contrasting phenotypes with regards to susceptibility and immune responses to fungal entomopathogenic infection in these two mosquitoes. This study provides new insights into the intricate multipartite interaction between the mosquito host, its native symbiont and pathogenic microbes that might be employed to control mosquito populations.}, } @article {pmid34842379, year = {2021}, author = {Victoria Rombot, D and Yermia Semuel, M}, title = {The Metagenomic Analysis of Potential Pathogenic Emerging Bacteria in Fleas.}, journal = {Pakistan journal of biological sciences : PJBS}, volume = {24}, number = {10}, pages = {1084-1090}, doi = {10.3923/pjbs.2021.1084.1090}, pmid = {34842379}, issn = {1812-5735}, mesh = {Animals ; Bacteria/*genetics/pathogenicity ; Metagenomics/methods ; Pesticides ; Siphonaptera/*metabolism/microbiology ; }, abstract = {Background and Objective: At present many pathogenic microbes that cause disease in humans are transmitted through animals. Ctenocephalides felisis specific ectoparasites in cats. Metagenomic research on the digestive tract and body surface of C. felishas been conducted. DNA genomics was extracted from the body surface and digestive tract of C. felis. Materials and Methods: Metagenomic analysis has used the 16S rRNA gene (V3-V4 region). Sequencing was carried out using New Generation Sequencing at the First BASE Laboratory, Singapore. Results: Wolbachia has the most significant bacterial composition in C. felis (94.4%), we were found bacteria with a composition >1% that have never been reported to be associated with C. felis. Also, there were 0.2% of bacteria whose taxonomic status cannot be determined. Conclusion: The results of this study become a vital reference pathogenic bacteria that can be transmitted to humans and animals through C. felis. It is necessary to study the resistance of bacteria isolated fromC. felisto antibiotics in the future.}, } @article {pmid34838119, year = {2021}, author = {Hamlili, FZ and Bérenger, JM and Diarra, AZ and Parola, P}, title = {Molecular and MALDI-TOF MS identification of swallow bugs Cimex hirundinis (Heteroptera: Cimicidae) and endosymbionts in France.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {587}, pmid = {34838119}, issn = {1756-3305}, mesh = {Animals ; *Bedbugs/classification/microbiology ; Birds ; France ; Humans ; Pathology, Molecular ; Phylogeny ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods ; Swallows/*parasitology ; Symbiosis ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: The Cimicidae are obligatory blood-feeding ectoparasites of medical and veterinary importance. We aim in the current study to assess the ability of MALDI-TOF MS to identify Cimex hirundinis swallow bugs collected in house martin nests.

METHODS: Swallow bugs were picked out from abandoned nests of house martin swallows and identified morphologically to the species level. The bugs were randomly selected, dissected and then subjected to MALDI-TOF MS and molecular analyses.

RESULTS: A total of 65 adults and 50 nymphs were used in the attempt to determine whether this tool could identify the bug species and discriminate their developmental stages. Five adults and four nymphs of C. hirundinis specimens were molecularly identified to update our MS homemade arthropod database. BLAST analysis of COI gene sequences from these C. hirundinis revealed 98.66-99.12% identity with the corresponding sequences of C. hirundinis of the GenBank. The blind test against the database supplemented with MS reference spectra showed 100% (57/57) C. hirundinis adults and 100% (46/46) C. hirundinis nymphs were reliably identified and in agreement with morphological identification with logarithmic score values between 1.922 and 2.665. Ninety-nine percent of C. hirundinis specimens tested were positive for Wolbachia spp. The sequencing results revealed that they were identical to Wolbachia massiliensis, belonging to the new T-supergroup strain and previously isolated from C. hemipterus.

CONCLUSIONS: We report for the first time to our knowledge a case of human infestation by swallow bugs (C. hirundinis) in France. We also show the usefulness of MALDI-TOF MS in the rapid identification of C. hirundinis specimens and nymphs with minimal sample requirements. We phylogenetically characterized the novel Wolbachia strain (W. massiliensis) infecting C. hirundinis and compared it to other recognized Wolbachia clades.}, } @article {pmid34831751, year = {2021}, author = {Soh, LT and Ong, Z and Vasquez, K and Chen, I and Li, X and Niah, W and Panchapakesan, C and Sheldenkar, A and Sim, S and Ng, LC and Lwin, MO}, title = {A Household-Based Survey to Understand Factors Influencing Awareness, Attitudes and Knowledge towards Wolbachia-Aedes Technology.}, journal = {International journal of environmental research and public health}, volume = {18}, number = {22}, pages = {}, pmid = {34831751}, issn = {1660-4601}, mesh = {*Aedes ; Animals ; Attitude ; *Dengue ; Male ; Mosquito Vectors ; Technology ; *Wolbachia ; }, abstract = {In 2016, Singapore introduced the release of male Wolbachia-Aedes mosquitoes to complement vector control efforts and suppress Aedes aegypti mosquitoes in selected study sites. With ongoing expansion of Project Wolbachia-Singapore to cover larger areas, a household-based survey was conducted between July 2019 to February 2020 in two Project Wolbachia study sites using a structured questionnaire, to evaluate current sentiments and assess the need for enhanced public messaging and engagement. The association of factors that influence awareness, attitudes, and knowledge towards the use of Wolbachia-Aedes technology was analysed using Pearson's Chi-square test and binary logistic regression. Of 500 respondents, 74.8% were aware of Project Wolbachia-Singapore. Comparatively, the level of knowledge on Wolbachia-Aedes technology was lower, suggesting knowledge gaps that require enhanced communication and messaging to address misinformation. Longer exposure to the project predicted greater awareness, whereas higher education levels predicted higher knowledge levels. Younger age groups and higher education levels were associated with high acceptance towards the project. High levels of trust and acceptance towards the project were also observed across the population. The public's positive perception of the project is a testament to the effective public communication undertaken to date and will facilitate programme expansion.}, } @article {pmid34829911, year = {2021}, author = {Kwofie, SK and Broni, E and Yunus, FU and Nsoh, J and Adoboe, D and Miller, WA and Wilson, MD}, title = {Molecular Docking Simulation Studies Identifies Potential Natural Product Derived-Antiwolbachial Compounds as Filaricides against Onchocerciasis.}, journal = {Biomedicines}, volume = {9}, number = {11}, pages = {}, pmid = {34829911}, issn = {2227-9059}, abstract = {Onchocerciasis is the leading cause of blindness and severe skin lesions which remain a major public health problem, especially in tropical areas. The widespread use of antibiotics and the long duration required for effective treatment continues to add to the increasing global menace of multi-resistant pathogens. Onchocerca volvulus harbors the endosymbiont bacteria Wolbachia, essential for the normal development of embryos, larvae and long-term survival of the adult worm, O. volvulus. We report here results of using structure-based drug design (SBDD) approach aimed at identifying potential novel Wolbachia inhibitors from natural products against the Wolbachia surface protein (WSP). The protein sequence of the WSP with UniProtKB identifier Q0RAI4 was used to model the three-dimensional (3D) structure via homology modelling techniques using three different structure-building algorithms implemented in Modeller, I-TASSER and Robetta. Out of the 15 generated models of WSP, one was selected as the most reasonable quality model which had 82, 15.5, 1.9 and 0.5% of the amino acid residues in the most favored regions, additionally allowed regions, generously allowed regions and disallowed regions, respectively, based on the Ramachandran plot. High throughput virtual screening was performed via Autodock Vina with a library comprising 42,883 natural products from African and Chinese databases, including 23 identified anti-Onchocerca inhibitors. The top six compounds comprising ZINC000095913861, ZINC000095486235, ZINC000035941652, NANPDB4566, acetylaleuritolic acid and rhemannic acid had binding energies of -12.7, -11.1, -11.0, -11, -10.3 and -9.5 kcal/mol, respectively. Molecular dynamics simulations including molecular mechanics Poisson-Boltzmann (MMPBSA) calculations reinforced the stability of the ligand-WSP complexes and plausible binding mechanisms. The residues Arg45, Tyr135, Tyr148 and Phe195 were predicted as potential novel critical residues required for ligand binding in pocket 1. Acetylaleuritolic acid and rhemannic acid (lantedene A) have previously been shown to possess anti-onchocercal activity. This warrants the need to evaluate the anti-WSP activity of the identified molecules. The study suggests the exploitation of compounds which target both pockets 1 and 2, by investigating their potential for effective depletion of Wolbachia. These compounds were predicted to possess reasonably good pharmacological profiles with insignificant toxicity and as drug-like. The compounds were computed to possess biological activity including antibacterial, antiparasitic, anthelmintic and anti-rickettsials. The six natural products are potential novel antiwolbachial agents with insignificant toxicities which can be explored further as filaricides for onchocerciasis.}, } @article {pmid34824354, year = {2021}, author = {Cogni, R and Ding, SD and Pimentel, AC and Day, JP and Jiggins, FM}, title = {Wolbachia reduces virus infection in a natural population of Drosophila.}, journal = {Communications biology}, volume = {4}, number = {1}, pages = {1327}, pmid = {34824354}, issn = {2399-3642}, support = {NAF\R1\180244//Royal Society/ ; }, mesh = {Animals ; Connecticut ; Drosophila melanogaster/*microbiology/*virology ; Male ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a maternally transmitted bacterial symbiont that is estimated to infect approximately half of arthropod species. In the laboratory it can increase the resistance of insects to viral infection, but its effect on viruses in nature is unknown. Here we report that in a natural population of Drosophila melanogaster, individuals that are infected with Wolbachia are less likely to be infected by viruses. By characterising the virome by metagenomic sequencing and then testing individual flies for infection, we found the protective effect of Wolbachia was virus-specific, with the prevalence of infection being up to 15% greater in Wolbachia-free flies. The antiviral effects of Wolbachia may contribute to its extraordinary ecological success, and in nature the symbiont may be an important component of the antiviral defences of insects.}, } @article {pmid34823581, year = {2021}, author = {Bleidorn, C and Henze, K}, title = {A new primer pair for barcoding of bees (Hymenoptera: Anthophila) without amplifying the orthologous coxA gene of Wolbachia bacteria.}, journal = {BMC research notes}, volume = {14}, number = {1}, pages = {427}, pmid = {34823581}, issn = {1756-0500}, mesh = {Animals ; Bacteria ; Bees/genetics ; DNA Barcoding, Taxonomic ; Hip ; *Hymenoptera ; *Wolbachia/genetics ; }, abstract = {OBJECTIVES: DNA barcoding became an effective method for the identification and monitoring of bees. However, standard primer pairs used for barcoding often result in (co-) amplification of bacterial endosymbionts of the genus Wolbachia, which are widespread among bee species. Here we designed a new primer pair and compared it with the performance of the standard Folmer-primers for a small sample set of bees representing the main taxonomic groups of bees.

RESULTS: The newly designed primer pair (BeeCox1F1/BeeCox1R2) outperformed the standard barcoding primer (LCO1490/HCO2198). By generating barcodes for a small test set of bees we found that the new primer pair produced high-quality sequences in all cases for unambiguous species identification using BOLD. Conversely, the standard barcoding primers often co-amplified the homologous Wolbachia gene and resulted in mixed chromatogram signals. These sequences showed high similarity with the bacterial endosymbiont instead of the host.}, } @article {pmid34820166, year = {2021}, author = {Price, DC and Brennan, JR and Wagner, NE and Egizi, AM}, title = {Comparative hologenomics of two Ixodes scapularis tick populations in New Jersey.}, journal = {PeerJ}, volume = {9}, number = {}, pages = {e12313}, pmid = {34820166}, issn = {2167-8359}, abstract = {Tick-borne diseases, such as those transmitted by the blacklegged tick Ixodes scapularis, are a significant and growing public health problem in the US. There is mounting evidence that co-occurring non-pathogenic microbes can also impact tick-borne disease transmission. Shotgun metagenome sequencing enables sampling of the complete tick hologenome-the collective genomes of the tick and all of the microbial species contained therein, whether pathogenic, commensal or symbiotic. This approach simultaneously uncovers taxonomic composition and allows the detection of intraspecific genetic variation, making it a useful tool to compare spatial differences across tick populations. We evaluated this approach by comparing hologenome data from two tick samples (N = 6 ticks per location) collected at a relatively fine spatial scale, approximately 23 km apart, within a single US county. Several intriguing variants in the data between the two sites were detected, including polymorphisms in both in the tick's own mitochondrial DNA and that of a rickettsial endosymbiont. The two samples were broadly similar in terms of the microbial species present, including multiple known tick-borne pathogens (Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum), filarial nematodes, and Wolbachia and Babesia species. We assembled the complete genome of the rickettsial endosymbiont (most likely Rickettsia buchneri) from both populations. Our results provide further evidence for the use of shotgun metagenome sequencing as a tool to compare tick hologenomes and differentiate tick populations across localized spatial scales.}, } @article {pmid34819638, year = {2021}, author = {Adams, KL and Abernathy, DG and Willett, BC and Selland, EK and Itoe, MA and Catteruccia, F}, title = {Wolbachia cifB induces cytoplasmic incompatibility in the malaria mosquito vector.}, journal = {Nature microbiology}, volume = {6}, number = {12}, pages = {1575-1582}, pmid = {34819638}, issn = {2058-5276}, support = {/HHMI/Howard Hughes Medical Institute/United States ; }, mesh = {Aedes/genetics/microbiology/physiology ; Animals ; Anopheles/genetics/*microbiology/*physiology ; Bacterial Proteins/genetics/*metabolism ; *Extrachromosomal Inheritance ; Female ; Infertility, Male ; Malaria/transmission ; Male ; Mosquito Vectors/genetics/*microbiology/*physiology ; Wolbachia/genetics/*metabolism ; }, abstract = {Wolbachia, a maternally inherited intracellular bacterial species, can manipulate host insect reproduction by cytoplasmic incompatibility (CI), which results in embryo lethality in crosses between infected males and uninfected females. CI is encoded by two prophage genes, cifA and cifB. Wolbachia, coupled with the sterile insect technique, has been used in field trials to control populations of the dengue vector Aedes albopictus, but CI-inducing strains are not known to infect the malaria vector Anopheles gambiae. Here we show that cifA and cifB can induce conditional sterility in the malaria vector An. gambiae. We used transgenic expression of these Wolbachia-derived genes in the An. gambiae germline to show that cifB is sufficient to cause embryonic lethality and that cifB-induced sterility is rescued by cifA expression in females. When we co-expressed cifA and cifB in male mosquitoes, the CI phenotype was attenuated. In female mosquitoes, cifB impaired fertility, which was overcome by co-expression of cifA. Our findings pave the way towards using CI to control malaria mosquito vectors.}, } @article {pmid34792280, year = {2022}, author = {Sanaei, E and Lin, YP and Cook, LG and Engelstädter, J}, title = {Wolbachia in scale insects: a distinct pattern of infection frequencies and potential transfer routes via ant associates.}, journal = {Environmental microbiology}, volume = {24}, number = {3}, pages = {1326-1339}, doi = {10.1111/1462-2920.15833}, pmid = {34792280}, issn = {1462-2920}, mesh = {Animals ; *Ants ; *Arthropods ; Biological Evolution ; *Hemiptera ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia is one of the most successful endosymbiotic bacteria of arthropods. Known as the 'master of manipulation', Wolbachia can induce a wide range of phenotypes in its host that can have far-reaching ecological and evolutionary consequences and may be exploited for disease and pest control. However, our knowledge of Wolbachia's distribution and the infection rate is unevenly distributed across arthropod groups such as scale insects. We fitted a distribution of within-species prevalence of Wolbachia to our data and compared it to distributions fitted to an up-to-date dataset compiled from surveys across all arthropods. The estimated distribution parameters indicate a Wolbachia infection frequency of 43.6% (at a 10% prevalence threshold) in scale insects. Prevalence of Wolbachia in scale insects follows a distribution similar to exponential decline (most species are predicted to have low prevalence infections), in contrast to the U-shaped distribution estimated for other taxa (most species have a very low or very high prevalence). We observed no significant associations between Wolbachia infection and scale insect traits. Finally, we screened for Wolbachia in scale insect's ecological associates. We found a positive correlation between Wolbachia infection in scale insects and their ant associates, pointing to a possible route of horizontal transfer of Wolbachia.}, } @article {pmid34791181, year = {2022}, author = {Mostoufi, SL and Singh, ND}, title = {Diet-induced changes in titer support a discrete response of Wolbachia-associated plastic recombination in Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {12}, number = {1}, pages = {}, pmid = {34791181}, issn = {2160-1836}, support = {T32 GM007413/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/physiology ; Phenotype ; Plastics ; Recombination, Genetic ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Plastic recombination in Drosophila melanogaster has been associated with a variety of extrinsic and intrinsic factors such as temperature, starvation, and parasite infection. The bacterial endosymbiont Wolbachia pipientis has also been associated with plastic recombination in D. melanogaster. Wolbachia infection is pervasive in arthropods and this infection induces a variety of phenotypes in its hosts, the strength of which can depend on bacterial titer. Here, we test the hypothesis that the magnitude of Wolbachia-associated plastic recombination in D. melanogaster depends on titer. To manipulate titer, we raised Wolbachia-infected and uninfected flies on diets that have previously been shown to increase or decrease Wolbachia titer relative to controls. We measured recombination in treated and control individuals using a standard backcrossing scheme with two X-linked visible markers. Our results recapitulate previous findings that Wolbachia infection is associated with increased recombination rate across the yellow-vermillion interval of the X chromosome. Our data show no significant effect of diet or diet by Wolbachia interactions on recombination, suggesting that diet-induced changes in Wolbachia titer have no effect on the magnitude of plastic recombination. These findings represent one of the first steps toward investigating Wolbachia-associated plastic recombination and demonstrate that the phenotype is a discrete response rather than a continuous one.}, } @article {pmid34765121, year = {2021}, author = {Gupta, M and Kaur, R and Gupta, A and Raychoudhury, R}, title = {Are ecological communities the seat of endosymbiont horizontal transfer and diversification? A case study with soil arthropod community.}, journal = {Ecology and evolution}, volume = {11}, number = {21}, pages = {14490-14508}, pmid = {34765121}, issn = {2045-7758}, abstract = {Maternally inherited endosymbionts of arthropods are one of the most abundant and diverse group of bacteria. These bacterial endosymbionts also show extensive horizontal transfer to taxonomically unrelated hosts and widespread recombination in their genomes. Such horizontal transfers can be enhanced when different arthropod hosts come in contact like in an ecological community. Higher rates of horizontal transfer can also increase the probability of recombination between endosymbionts, as they now share the same host cytoplasm. However, reports of community-wide endosymbiont data are rare as most studies choose few host taxa and specific ecological interactions among the hosts. To better understand endosymbiont spread within host populations, we investigated the incidence, diversity, extent of horizontal transfer, and recombination of three endosymbionts (Wolbachia, Cardinium, and Arsenophonus) in a specific soil arthropod community. Wolbachia strains were characterized with MLST genes whereas 16S rRNA gene was used for Cardinium and Arsenophonus. Among 3,509 individual host arthropods, belonging to 390 morphospecies, 12.05% were infected with Wolbachia, 2.82% with Cardinium and 2.05% with Arsenophonus. Phylogenetic incongruence between host and endosymbiont indicated extensive horizontal transfer of endosymbionts within this community. Three cases of recombination between Wolbachia supergroups and eight incidences of within-supergroup recombination were also found. Statistical tests of similarity indicated supergroup A Wolbachia and Cardinium show a pattern consistent with extensive horizontal transfer within the community but not for supergroup B Wolbachia and Arsenophonus. We highlight the importance of extensive community-wide studies for a better understanding of the spread of endosymbionts across global arthropod communities.}, } @article {pmid34749528, year = {2021}, author = {Mancini, MV and Ant, TH and Herd, CS and Martinez, J and Murdochy, SM and Gingell, DD and Mararo, E and Johnson, PCD and Sinkins, SP}, title = {High Temperature Cycles Result in Maternal Transmission and Dengue Infection Differences Between Wolbachia Strains in Aedes aegypti.}, journal = {mBio}, volume = {12}, number = {6}, pages = {e0025021}, pmid = {34749528}, issn = {2150-7511}, support = {/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/growth & development/*microbiology/virology ; Animals ; Dengue/transmission/virology ; Dengue Virus/physiology ; Ecosystem ; Female ; Humans ; Larva/*growth & development/microbiology/virology ; Male ; Mosquito Control ; Mosquito Vectors/growth & development/*microbiology/virology ; Population Dynamics ; Temperature ; Wolbachia/genetics/*physiology ; }, abstract = {Environmental factors play a crucial role in the population dynamics of arthropod endosymbionts, and therefore in the deployment of Wolbachia symbionts for the control of dengue arboviruses. The potential of Wolbachia to invade, persist, and block virus transmission depends in part on its intracellular density. Several recent studies have highlighted the importance of larval rearing temperature in modulating Wolbachia densities in adults, suggesting that elevated temperatures can severely impact some strains, while having little effect on others. The effect of a replicated tropical heat cycle on Wolbachia density and levels of virus blocking was assessed using Aedes aegypti lines carrying strains wMel and wAlbB, two Wolbachia strains currently used for dengue control. Impacts on intracellular density, maternal transmission fidelity, and dengue inhibition capacity were observed for wMel. In contrast, wAlbB-carrying Ae. aegypti maintained a relatively constant intracellular density at high temperatures and conserved its capacity to inhibit dengue. Following larval heat treatment, wMel showed a degree of density recovery in aging adults, although this was compromised by elevated air temperatures. IMPORTANCE In the past decades, dengue incidence has dramatically increased all over the world. An emerging dengue control strategy utilizes Aedes aegypti mosquitoes artificially transinfected with the bacterial symbiont Wolbachia, with the ultimate aim of replacing wild mosquito populations. However, the rearing temperature of mosquito larvae is known to impact on some Wolbachia strains. In this study, we compared the effects of a temperature cycle mimicking natural breeding sites in tropical climates on two Wolbachia strains, currently used for open field trials. When choosing the Wolbachia strain to be used in a dengue control program it is important to consider the effects of environmental temperatures on invasiveness and virus inhibition. These results underline the significance of understanding the impact of environmental factors on released mosquitoes, in order to ensure the most efficient strategy for dengue control.}, } @article {pmid34725401, year = {2021}, author = {Rakotonirina, A and Caruzzo, C and Ballan, V and Kainiu, M and Marin, M and Colot, J and Richard, V and Dupont-Rouzeyrol, M and Selmaoui-Folcher, N and Pocquet, N}, title = {Wolbachia detection in Aedes aegypti using MALDI-TOF MS coupled to artificial intelligence.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {21355}, pmid = {34725401}, issn = {2045-2322}, mesh = {Aedes/*microbiology ; Animals ; Artificial Intelligence ; Mosquito Control/methods ; Mosquito Vectors/*microbiology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Wolbachia/chemistry/*isolation & purification ; }, abstract = {The mosquito Aedes aegypti is the major vector of arboviruses like dengue, Zika and chikungunya viruses. Attempts to reduce arboviruses emergence focusing on Ae. aegypti control has proven challenging due to the increase of insecticide resistances. An emerging strategy which consists of releasing Ae. aegypti artificially infected with Wolbachia in natural mosquito populations is currently being developed. The monitoring of Wolbachia-positive Ae. aegypti in the field is performed in order to ensure the program effectiveness. Here, the reliability of the Matrix‑Assisted Laser Desorption Ionization‑Time Of Flight (MALDI‑TOF) coupled with the machine learning methods like Convolutional Neural Network (CNN) to detect Wolbachia in field Ae. aegypti was assessed for the first time. For this purpose, laboratory reared and field Ae. aegypti were analyzed. The results showed that the CNN recognized Ae. aegypti spectral patterns associated with Wolbachia-infection. The MALDI-TOF coupled with the CNN (sensitivity = 93%, specificity = 99%, accuracy = 97%) was more efficient than the loop-mediated isothermal amplification (LAMP), and as efficient as qPCR for Wolbachia detection. It therefore represents an interesting method to evaluate the prevalence of Wolbachia in field Ae. aegypti mosquitoes.}, } @article {pmid34719095, year = {2022}, author = {Malkeyeva, D and Kiseleva, E and Fedorova, SA}, title = {Loss of Hsp67Bc leads to autolysosome enlargement in the Drosophila brain.}, journal = {Cell biology international}, volume = {46}, number = {2}, pages = {203-212}, doi = {10.1002/cbin.11721}, pmid = {34719095}, issn = {1095-8355}, support = {0259-2021-0011//Ministry of Science and Higher Education of the Russian Federation/ ; }, mesh = {Animals ; Brain/metabolism ; *Drosophila/genetics ; *Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster/metabolism ; Heat-Shock Proteins/metabolism ; Lysosomes/metabolism ; }, abstract = {Hsp67Bc is a small heat shock protein found in Drosophila melanogaster. Apart from performing a function (common for all small heat shock proteins) of preventing aggregation of misfolded proteins, it is involved in macroautophagy regulation alongside the Starvin protein. Overexpression of the D. melanogaster Hsp67Bc gene has been shown to stimulate macroautophagy in S2 cell culture. Nonetheless, it has been unknown how the absence of the Hsp67Bc gene may affect it. Here, we studied the effect of Hsp67Bc gene deletion on the macroautophagy induced by the pathogenic Wolbachia wMelPop strain in D. melanogaster. We detected Wolbachia inside autophagic vacuoles in fly neurons, thereby proving that these endosymbionts were being eliminated via macroautophagy. Nevertheless, we did not register any difference in brain bacterial load between Hsp67Bc-null and control flies at all tested stages of ontogenesis. Moreover, the abundance of autophagic vacuoles was similar between neurons of the mutant and control flies, yet the cross-sectional area of autolysosomes on ultrathin sections was more than 1.5-fold larger in Hsp67Bc-null fly brains than in the control line. Our findings suggest that the product of the Hsp67Bc gene does not participate in the initiation of endosymbiont-induced macroautophagy but may mediate autophagosome maturation: the deletion of the Hsp67Bc gene leads to the increase in autolysosome size.}, } @article {pmid34717929, year = {2021}, author = {Prichard, RK}, title = {Macrocyclic lactone resistance in Dirofilaria immitis: risks for prevention of heartworm disease.}, journal = {International journal for parasitology}, volume = {51}, number = {13-14}, pages = {1121-1132}, doi = {10.1016/j.ijpara.2021.08.006}, pmid = {34717929}, issn = {1879-0135}, mesh = {Animals ; *Cat Diseases/drug therapy ; Cats ; *Dirofilaria immitis/genetics ; *Dirofilariasis/prevention & control ; *Dog Diseases/parasitology/prevention & control ; Dogs ; Genetic Markers ; Lactones/pharmacology/therapeutic use ; Microfilariae ; }, abstract = {Heartworm disease, caused by Dirofilaria immitis, can be lethal in dogs and cats. It is transmitted by mosquitoes, and occurs in many parts of the world. Prevention relies on macrocyclic lactones. Macrocyclic lactones used are ivermectin, selamectin, abamectin, eprinomectin, milbemycin oxime and moxidectin, administered at 30-day intervals during the transmission season. Some moxidectin formulations are long-acting injectables. In the USA, preventives are recommended throughout the year. Loss of efficacy of macrocyclic lactone preventives was reported in 2005 and proof of resistance in the USA was published a decade later. Understanding factors which promote resistance is important to maintain control. Factors important for resistance development are discussed. Better, inexpensive tests to confirm resistance are needed. Infection in animals under chemoprophylaxis per se does not imply resistance because lack of compliance in preventive use could be the reason. In vivo confirmation of resistance is expensive, slow and ethically questionable. A microfilariae suppression test can be a surrogate test, but requires a high dose of a macrocyclic lactone and repeated blood microfilaria counts 2-4 weeks later. DNA single nucleotide polymorphism markers have been successfully used. However, the specific genetic changes which cause resistance are unknown. Surveys to map and follow the extent of resistance are needed. Long acting mosquito repellants and insecticides can play a useful role. High dose rate formulations of moxidectin, coupled with mosquito biting mitigation may reduce transmission of resistant genotypes. Doxycycline, daily for 28 days, as anti-Wolbachia treatment, can reduce transmission and remove adult parasites. However, new classes of heartworm preventives are needed. While any preventive strategy must be highly effective, registration requirements for 100% efficacy may hinder development of useful new classes of preventives. Continued reliance on macrocyclic lactone preventives, when they do not work against resistant genotypes, will spread resistance, and allow for more disease.}, } @article {pmid34710095, year = {2021}, author = {Yang, Y and He, Y and Zhu, G and Zhang, J and Gong, Z and Huang, S and Lu, G and Peng, Y and Meng, Y and Hao, X and Wang, C and Sun, J and Shang, S}, title = {Prevalence and molecular characterization of Wolbachia in field-collected Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus in China.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {10}, pages = {e0009911}, pmid = {34710095}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Anopheles/*microbiology ; China ; Culex/*microbiology ; Female ; Male ; Mosquito Vectors/microbiology ; Multilocus Sequence Typing ; Phylogeny ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China.}, } @article {pmid34704919, year = {2021}, author = {Ortiz-Baez, AS and Shi, M and Hoffmann, AA and Holmes, EC}, title = {RNA virome diversity and Wolbachia infection in individual Drosophila simulans flies.}, journal = {The Journal of general virology}, volume = {102}, number = {10}, pages = {}, pmid = {34704919}, issn = {1465-2099}, mesh = {Animals ; Drosophila simulans/*microbiology/virology ; Female ; Phylogeny ; RNA Viruses/classification/genetics/isolation & purification/*physiology ; Symbiosis ; Virome/genetics/*physiology ; Wolbachia/isolation & purification/*physiology ; }, abstract = {The endosymbiont bacteria of the genus Wolbachia are associated with multiple mutualistic effects on insect biology, including nutritional and antiviral properties. Members of the genus Wolbachia naturally occur in fly species of the genus Drosophila, providing an operational model host for studying how virome composition may be affected by its presence. Drosophila simulans populations can carry a variety of strains of members of the genus Wolbachia, with the wAu strain associated with strong antiviral protection under experimental conditions. We used D. simulans sampled from the Perth Hills, Western Australia, to investigate the potential virus protective effect of the wAu strain of Wolbachia on individual wild-caught flies. Our data revealed no appreciable variation in virus composition and abundance between individuals infected or uninfected with Wolbachia associated with the presence or absence of wAu. However, it remains unclear whether wAu might affect viral infection and host survival by increasing tolerance rather than inducing complete resistance. These data also provide new insights into the natural virome diversity of D. simulans. Despite the small number of individuals sampled, we identified a repertoire of RNA viruses, including nora virus, galbut virus, thika virus and La Jolla virus, that have been identified in other species of the genus Drosophila. Chaq virus-like sequences associated with galbut virus were also detected. In addition, we identified five novel viruses from the families Reoviridae, Tombusviridae, Mitoviridae and Bunyaviridae. Overall, this study highlights the complex interaction between Wolbachia and RNA virus infections and provides a baseline description of the natural virome of D. simulans.}, } @article {pmid34680640, year = {2021}, author = {Bell-Sakyi, L and Beliavskaia, A and Hartley, CS and Jones, L and Luu, L and Haines, LR and Hamilton, JGC and Darby, AC and Makepeace, BL}, title = {Isolation in Natural Host Cell Lines of Wolbachia Strains wPip from the Mosquito Culex pipiens and wPap from the Sand Fly Phlebotomus papatasi.}, journal = {Insects}, volume = {12}, number = {10}, pages = {}, pmid = {34680640}, issn = {2075-4450}, support = {BBS/E/I/00007039/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BBS/E/I/00002118/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 204806/Z/16/Z and 080961/Z/06/Z/WT_/Wellcome Trust/United Kingdom ; BB/P024270/1, BBS/E/I/00002118 and BBS/E/I/00007039/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, abstract = {Endosymbiotic intracellular bacteria of the genus Wolbachia are harboured by many species of invertebrates. They display a wide range of developmental, metabolic and nutritional interactions with their hosts and may impact the transmission of arboviruses and protozoan parasites. Wolbachia have occasionally been isolated during insect cell line generation. Here, we report the isolation of two strains of Wolbachia, wPip and wPap, during cell line generation from their respective hosts, the mosquito Culex pipiens and the sand fly Phlebotomus papatasi. wPip was pathogenic for both new C. pipiens cell lines, CPE/LULS50 and CLP/LULS56, requiring tetracycline treatment to rescue the lines. In contrast, wPap was tolerated by the P. papatasi cell line PPL/LULS49, although tetracycline treatment was applied to generate a Wolbachia-free subline. Both Wolbachia strains were infective for a panel of heterologous insect and tick cell lines, including two novel lines generated from the sand fly Lutzomyia longipalpis, LLE/LULS45 and LLL/LULS52. In all cases, wPip was more pathogenic for the host cells than wPap. These newly isolated Wolbachia strains, and the novel mosquito and sand fly cell lines reported here, will add to the resources available for research on host-endosymbiont relationships, as well as on C. pipiens, P. papatasi, L. longipalpis and the pathogens that they transmit.}, } @article {pmid34680622, year = {2021}, author = {Shapoval, NA and Nokkala, S and Nokkala, C and Kuftina, GN and Kuznetsova, VG}, title = {The Incidence of Wolbachia Bacterial Endosymbiont in Bisexual and Parthenogenetic Populations of the Psyllid Genus Cacopsylla (Hemiptera, Psylloidea).}, journal = {Insects}, volume = {12}, number = {10}, pages = {}, pmid = {34680622}, issn = {2075-4450}, support = {19-14-00202//Russian Science Foundation/ ; }, abstract = {Wolbachia is one of the most common intracellular bacteria; it infects a wide variety of insects, other arthropods, and some nematodes. Wolbachia is ordinarily transmitted vertically from mother to offspring and can manipulate physiology and reproduction of their hosts in different ways, e.g., induce feminization, male killing, and parthenogenesis. Despite the great interest in Wolbachia, many aspects of its biology remain unclear and its incidence across many insect orders, including Hemiptera, is still poorly understood. In this report, we present data on Wolbachia infection in five jumping plant-lice species (Hemiptera, Psylloidea) of the genus Cacopsylla Ossiannilsson, 1970 with different reproductive strategies and test the hypothesis that Wolbachia mediates parthenogenetic and bisexual patterns observed in some Cacopsylla species. We show that the five species studied are infected with a single Wolbachia strain, belonging to the supergroup B. This strain has also been found in different insect orders (Lepidoptera, Hemiptera, Plecoptera, Orthoptera, Hymenoptera, Diptera) and even in acariform mites (Trombidiformes), suggesting extensive horizontal transmission of Wolbachia between representatives of these taxa. Our survey did not reveal significant differences in infection frequency between parthenogenetic and bisexual populations or between males and females within bisexual populations. However, infection rate varied notably in different Cacopsylla species or within distinct populations of the same species. Overall, we demonstrate that Wolbachia infects a high proportion of Cacopsylla individuals and populations, suggesting the essential role of this bacterium in their biology.}, } @article {pmid34677126, year = {2021}, author = {Perlmutter, JI and Meyers, JE and Bordenstein, SR}, title = {A single synonymous nucleotide change impacts the male-killing phenotype of prophage WO gene wmk.}, journal = {eLife}, volume = {10}, number = {}, pages = {}, pmid = {34677126}, issn = {2050-084X}, support = {R21 AI133522/AI/NIAID NIH HHS/United States ; F31 AI143152/AI/NIAID NIH HHS/United States ; P20 GM103418/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Drosophila melanogaster/*microbiology ; Male ; Microorganisms, Genetically-Modified/genetics/physiology ; Prophages/*genetics ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are the most widespread bacterial endosymbionts in animals. Within arthropods, these maternally transmitted bacteria can selfishly hijack host reproductive processes to increase the relative fitness of their transmitting females. One such form of reproductive parasitism called male killing, or the selective killing of infected males, is recapitulated to degrees by transgenic expression of the prophage WO-mediated killing (wmk) gene. Here, we characterize the genotype-phenotype landscape of wmk-induced male killing in D. melanogaster using transgenic expression. While phylogenetically distant wmk homologs induce no sex-ratio bias, closely-related homologs exhibit complex phenotypes spanning no death, male death, or death of all hosts. We demonstrate that alternative start codons, synonymous codons, and notably a single synonymous nucleotide in wmk can ablate killing. These findings reveal previously unrecognized features of transgenic wmk-induced killing and establish new hypotheses for the impacts of post-transcriptional processes in male killing variation. We conclude that synonymous sequence changes are not necessarily silent in nested endosymbiotic interactions with life-or-death consequences.}, } @article {pmid34666103, year = {2021}, author = {Poopandi, S and Sundaraj, R and Rajmichael, R and Thangaraj, S and Dhamodharan, P and Biswal, J and Malaisamy, V and Jeyaraj Pandian, C and Jeyaraman, J}, title = {Computational screening of potential inhibitors targeting MurF of Brugia malayi Wolbachia through multi-scale molecular docking, molecular dynamics and MM-GBSA analysis.}, journal = {Molecular and biochemical parasitology}, volume = {246}, number = {}, pages = {111427}, doi = {10.1016/j.molbiopara.2021.111427}, pmid = {34666103}, issn = {1872-9428}, mesh = {Animals ; *Brugia malayi ; *Elephantiasis, Filarial/parasitology ; Molecular Docking Simulation ; Molecular Dynamics Simulation ; *Wolbachia/metabolism ; }, abstract = {Lymphatic filariasis is a parasitic disease caused by the worms Wuchereria bancrofti, Brugia malayi and Brugia timori. Three anti-filarial drugs namely Diethylcarbamazine, Ivermectin and Albendazole and their combinations are used as the control strategies for filariasis. The disease has received much attention in drug discovery due to the unavailability of vaccines and the toxic pharmaceutical properties of the existing drugs. In Wolbachia endosymbiont Brugia malayi, the UDP-N-acetylmuramoyl-tripeptide-d-alanyl-d-alanine ligase (MurF) plays a key role in peptidoglycan biosynthesis pathway and therefore can be considered as effective drug target against filariasis disease. Therefore, in the present study, MurF was selected as the therapeutic target to identify specific inhibitors against filariasis. Homology modeling was performed to predict the three-dimensional structure of MurF due to the absence of the experimental structure. Further molecular dynamics simulation and structure-based high throughput virtual screening with three different chemical databases (Zinc, Maybridge and Specs) were carried out to identify potent inhibitors and also to check their conformations inside the binding site of MurF, respectively. Top three compounds with high docking score and high relative binding affinity against MurF were selected. Further, validation studies, including predicted ADME (Absorption, Distribution, Metabolism, Excretion) assessment, binding free energy using MM-GBSA (Molecular Mechanics Generalized Born Surface Area) and DFT (Density Functional Theory) calculations were performed for the top three compounds. From the results, it was observed that all the three compounds were predicted to show high reactivity, acceptable range of pharmacokinetic properties and high binding affinity with the drug target MurF. Overall, the results could provide more understanding on the inhibition of MurF enzyme and the screened compounds could lead to the development of new specific anti-filarial drugs.}, } @article {pmid34662426, year = {2022}, author = {Hill, T and Unckless, RL and Perlmutter, JI}, title = {Positive Selection and Horizontal Gene Transfer in the Genome of a Male-Killing Wolbachia.}, journal = {Molecular biology and evolution}, volume = {39}, number = {1}, pages = {}, pmid = {34662426}, issn = {1537-1719}, support = {P20 GM103418/GM/NIGMS NIH HHS/United States ; P20 GM103638/GM/NIGMS NIH HHS/United States ; R00 GM114714/GM/NIGMS NIH HHS/United States ; R01 AI139154/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila/genetics/microbiology ; Drosophila melanogaster/genetics ; Gene Transfer, Horizontal ; Genome ; Male ; *Wolbachia/genetics ; }, abstract = {Wolbachia are a genus of widespread bacterial endosymbionts in which some strains can hijack or manipulate arthropod host reproduction. Male killing is one such manipulation in which these maternally transmitted bacteria benefit surviving daughters in part by removing competition with the sons for scarce resources. Despite previous findings of interesting genome features of microbial sex ratio distorters, the population genomics of male-killers remain largely uncharacterized. Here, we uncover several unique features of the genome and population genomics of four Arizonan populations of a male-killing Wolbachia strain, wInn, that infects mushroom-feeding Drosophila innubila. We first compared the wInn genome with other closely related Wolbachia genomes of Drosophila hosts in terms of genome content and confirm that the wInn genome is largely similar in overall gene content to the wMel strain infecting D. melanogaster. However, it also contains many unique genes and repetitive genetic elements that indicate lateral gene transfers between wInn and non-Drosophila eukaryotes. We also find that, in line with literature precedent, genes in the Wolbachia prophage and Octomom regions are under positive selection. Of all the genes under positive selection, many also show evidence of recent horizontal transfer among Wolbachia symbiont genomes. These dynamics of selection and horizontal gene transfer across the genomes of several Wolbachia strains and diverse host species may be important underlying factors in Wolbachia's success as a male-killer of divergent host species.}, } @article {pmid34654483, year = {2021}, author = {Gloder, G and Bourne, ME and Verreth, C and Wilberts, L and Bossaert, S and Crauwels, S and Dicke, M and Poelman, EH and Jacquemyn, H and Lievens, B}, title = {Parasitism by endoparasitoid wasps alters the internal but not the external microbiome in host caterpillars.}, journal = {Animal microbiome}, volume = {3}, number = {1}, pages = {73}, pmid = {34654483}, issn = {2524-4671}, support = {FWO//Fonds Wetenschappelijk Onderzoek/ ; G.0961.19N//Fonds Wetenschappelijk Onderzoek/ ; ALWOP.343//Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ ; ALWOP.368//Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ ; }, abstract = {BACKGROUND: The microbiome of many insects consists of a diverse community of microorganisms that can play critical roles in the functioning and overall health of their hosts. Although the microbial communities of insects have been studied thoroughly over the past decade, little is still known about how biotic interactions affect the microbial community structure in and on the bodies of insects. In insects that are attacked by parasites or parasitoids, it can be expected that the microbiome of the host insect is affected by the presence of these parasitic organisms that develop in close association with their host. In this study, we used high-throughput amplicon sequencing targeting both bacteria and fungi to test the hypothesis that parasitism by the endoparasitoid Cotesia glomerata affected the microbiome of its host Pieris brassicae. Healthy and parasitized caterpillars were collected from both natural populations and a laboratory culture.

RESULTS: Significant differences in bacterial community structure were found between field-collected caterpillars and laboratory-reared caterpillars, and between the external and the internal microbiome of the caterpillars. Parasitism significantly altered the internal microbiome of caterpillars, but not the external microbiome. The internal microbiome of all parasitized caterpillars and of the parasitoid larvae in the caterpillar hosts was dominated by a Wolbachia strain, which was completely absent in healthy caterpillars, suggesting that the strain was transferred to the caterpillars during oviposition by the parasitoids.

CONCLUSION: We conclude that biotic interactions such as parasitism have pronounced effects on the microbiome of an insect host and possibly affect interactions with higher-order insects.}, } @article {pmid34650106, year = {2021}, author = {Wielkopolan, B and Krawczyk, K and Szabelska-Beręsewicz, A and Obrępalska-Stęplowska, A}, title = {The structure of the cereal leaf beetle (Oulema melanopus) microbiome depends on the insect's developmental stage, host plant, and origin.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {20496}, pmid = {34650106}, issn = {2045-2322}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Coleoptera/growth & development/*microbiology ; Hordeum ; Larva/microbiology ; *Microbiota ; RNA, Ribosomal, 16S ; Rickettsia/isolation & purification ; Triticum ; Wolbachia/isolation & purification ; }, abstract = {Cereal leaf beetle (CLB, Oulema melanopus, Coleoptera, Chrysomelidae) is a serious agricultural pest that causes considerable damages to agricultural production. The aim of this study was to characterize the bacterial communities associated with larvae and imagoes of CLB collected from various cereal host species and locations. The bacterial profile was characterized by 16S rRNA gene sequencing at the V3-V4 hypervariable region. Using taxonomy-based analysis, the bacterial community of CLB containing 16 phyla, 26 classes, 49 orders, 78 families, 94 genera, and 63 species of bacteria was identified. The abundance of Wolbachia, Rickettsia, and Lactococcus genus was significantly higher in CLB imagoes than in larvae. Statistical analysis confirmed that the bacterial community of the larvae is more diverse in comparison to imagoes and that insects collected from spring barley and wheat are characterized by a much higher biodiversity level of bacterial genera and species than insects collected from other cereals. Obtained results indicated that the developmental stage, the host plant, and the insect's sampling location affected the CLB's microbiome. Additionally, the CLB core microbiome was determined. It consists of 2 genera (Wolbachia and Rickettsia) shared by at least 90% tested CLB insects, regardless of the variables analysed.}, } @article {pmid34643449, year = {2021}, author = {Lefoulon, E and Truchon, A and Clark, T and Long, C and Frey, D and Slatko, BE}, title = {Greenhead (Tabanus nigrovittatus) Wolbachia and Its Microbiome: A Preliminary Study.}, journal = {Microbiology spectrum}, volume = {9}, number = {2}, pages = {e0051721}, pmid = {34643449}, issn = {2165-0497}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Diptera/*microbiology ; *Microbiota ; Phylogeny ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Endosymbiotic Wolbachia bacteria are known to influence the host physiology, microbiota composition, and dissemination of pathogens. We surveyed a population of Tabanus nigrovittatus, commonly referred to as "greenheads," from Crane Beach (Ipswich, MA, USA) for the presence of the alphaproteobacterial symbiont Wolbachia. We studied the COI (mitochondrial cytochrome oxidase) marker gene to evaluate the phylogenetic diversity of the studied specimens. The DNA sequences show strong similarity (between 99.9 and 98%) among the collected specimens but lower similarity to closely related entries in the NCBI database (only between 96.3 and 94.7%), suggesting a more distant relatedness. Low levels of Wolbachia presence necessitated a nested PCR approach, and using 5 markers (ftsZ, fbpA, dnaA, coxA, and gatB), we determined that two recognized "supergroups" of Wolbachia species were represented in the studied specimens, members of clades A and B. Using next-generation sequencing, we also surveyed the insect gut microbiomes of a subset of flies, using Illumina and PacBio 16S rRNA gene sequencing with barcoded primers. The composition of Proteobacteria also varied from fly to fly, with components belonging to Gammaproteobacteria making up the largest percentage of organisms (30 to 70%) among the microbiome samples. Most of the samples showed the presence of Spiroplasma, a member of the phylum Mollicutes, although the frequency of its presence was variable, ranging from 2 to 57%. Another noteworthy bacterial phylum consistently identified was Firmicutes, though the read abundances were typically below 10%. Of interest is an association between Wolbachia presence and higher Alphaproteobacteria representation in the microbiomes, suggesting that the presence of Wolbachia affects the host microbiome. IMPORTANCE Tabanus nigrovittatus greenhead populations contain two supergroups of Wolbachia endosymbionts, members of supergroups A and B. Analysis of the greenhead microbiome using next-generation sequencing revealed that the majority of bacterial species detected belonged to Gammaproteobacteria, with most of the samples also showing the presence of Spiroplasma, a member of the Mollicutes phylum also known to infect insects. An association between Wolbachia presence and higher Alphaproteobacteria representation in the microbiomes suggests that Wolbachia presence affects the host microbiome composition.}, } @article {pmid34642800, year = {2021}, author = {Ngwewondo, A and Scandale, I and Specht, S}, title = {Onchocerciasis drug development: from preclinical models to humans.}, journal = {Parasitology research}, volume = {120}, number = {12}, pages = {3939-3964}, pmid = {34642800}, issn = {1432-1955}, mesh = {Humans ; Neglected Diseases/drug therapy/prevention & control ; *Onchocerciasis/drug therapy ; *Pharmaceutical Preparations ; *Wolbachia ; }, abstract = {Twenty diseases are recognized as neglected tropical diseases (NTDs) by World Health Assembly resolutions, including human filarial diseases. The end of NTDs is embedded within the Sustainable Development Goals for 2030, under target 3.3. Onchocerciasis afflicts approximately 20.9 million people worldwide with > 90% of those infected residing in Africa. Control programs have made tremendous efforts in the management of onchocerciasis by mass drug administration and aerial larviciding; however, disease elimination is not yet achieved. In the new WHO roadmap, it is recognized that new drugs or drug regimens that kill or permanently sterilize adult filarial worms would significantly improve elimination timelines and accelerate the achievement of the program goal of disease elimination. Drug development is, however, handicapped by high attrition rates, and many promising molecules fail in preclinical development or in subsequent toxicological, safety and efficacy testing; thus, research and development (R&D) costs are, in aggregate, very high. Drug discovery and development for NTDs is largely driven by unmet medical needs put forward by the global health community; the area is underfunded and since no high return on investment is possible, there is no dedicated drug development pipeline for human filariasis. Repurposing existing drugs is one approach to filling the drug development pipeline for human filariasis. The high cost and slow pace of discovery and development of new drugs has led to the repurposing of "old" drugs, as this is more cost-effective and allows development timelines to be shortened. However, even if a drug is marketed for a human or veterinary indication, the safety margin and dosing regimen will need to be re-evaluated to determine the risk in humans. Drug repurposing is a promising approach to enlarging the pool of active molecules in the drug development pipeline. Another consideration when providing new treatment options is the use of combinations, which is not addressed in this review. We here summarize recent advances in the late preclinical or early clinical stage in the search for a potent macrofilaricide, including drugs against the nematode and against its endosymbiont, Wolbachia pipientis.}, } @article {pmid36962069, year = {2021}, author = {Soh, S and Ho, SH and Seah, A and Ong, J and Dickens, BS and Tan, KW and Koo, JR and Cook, AR and Tan, KB and Sim, S and Ng, LC and Lim, JT}, title = {Economic impact of dengue in Singapore from 2010 to 2020 and the cost-effectiveness of Wolbachia interventions.}, journal = {PLOS global public health}, volume = {1}, number = {10}, pages = {e0000024}, pmid = {36962069}, issn = {2767-3375}, abstract = {The release of Wolbachia-infected mosquitoes is a promising disease intervention strategy that aims to control dengue and other arboviral infections. While early field trials and modelling studies suggest promising epidemiological and entomological outcomes, the overall cost effectiveness of the technology is not well studied in a resource rich setting nor under the suppression approach that aims to suppress the wild-type mosquito population through the release of Wolbachia-infected males. We used economical and epidemiological data from 2010 to 2020 to first ascertain the economic and health costs of dengue in Singapore, a high income nation where dengue is hyper-endemic. The hypothetical cost effectiveness of a national Wolbachia suppression program was then evaluated historically from 2010 to 2020. We estimated that the average economic impact of dengue in Singapore from 2010 to 2020 in constant 2010US$ ranged from $1.014 to $2.265 Billion. Using empirically derived disability weights, we estimated a disease burden of 7,645-21,262 DALYs from 2010-2020. Under an assumed steady-state running cost of a national Wolbachia suppression program in Singapore, we conservatively estimate that Wolbachia would cost an estimated $50,453-$100,907 per DALYs averted and would lead to an estimated $329.40 Million saved in economic costs over 2010 to 2020 under 40% intervention efficacy. Wolbachia releases in Singapore are expected to be highly cost-effective and its rollout must be prioritised to reduce the onward spread of dengue.}, } @article {pmid34634928, year = {2021}, author = {Jiménez, NE and Gerdtzen, ZP and Olivera-Nappa, Á and Salgado, JC and Conca, C}, title = {Novel Symbiotic Genome-Scale Model Reveals Wolbachia's Arboviral Pathogen Blocking Mechanism in Aedes aegypti.}, journal = {mBio}, volume = {12}, number = {5}, pages = {e0156321}, pmid = {34634928}, issn = {2150-7511}, mesh = {Aedes/*microbiology/*virology ; Amino Acids/metabolism ; Animals ; Arboviruses/metabolism/*pathogenicity ; *Genome, Bacterial ; Host Microbial Interactions ; Lipid Metabolism ; Mosquito Vectors/microbiology/virology ; Symbiosis/*genetics ; Virus Replication/physiology ; Wolbachia/*genetics/metabolism/*virology ; }, abstract = {Wolbachia are endosymbiont bacteria known to infect arthropods causing different effects, such as cytoplasmic incompatibility and pathogen blocking in Aedes aegypti. Although several Wolbachia strains have been studied, there is little knowledge regarding the relationship between this bacterium and their hosts, particularly on their obligate endosymbiont nature and its pathogen blocking ability. Motivated by the potential applications on disease control, we developed a genome-scale model of two Wolbachia strains: wMel and the strongest Dengue blocking strain known to date: wMelPop. The obtained metabolic reconstructions exhibit an energy metabolism relying mainly on amino acids and lipid transport to support cell growth that is consistent with altered lipid and cholesterol metabolism in Wolbachia-infected mosquitoes. The obtained metabolic reconstruction was then coupled with a reconstructed mosquito model to retrieve a symbiotic genome-scale model accounting for 1,636 genes and 6,408 reactions of the Aedes aegypti-Wolbachia interaction system. Simulation of an arboviral infection in the obtained novel symbiotic model represents a metabolic scenario characterized by pathogen blocking in higher titer Wolbachia strains, showing that pathogen blocking by Wolbachia infection is consistent with competition for lipid and amino acid resources between arbovirus and this endosymbiotic bacteria. IMPORTANCE Arboviral diseases such as Zika and Dengue have been on the rise mainly due to climate change, and the development of new treatments and strategies to limit their spreading is needed. The use of Wolbachia as an approach for disease control has motivated new research related to the characterization of the mechanisms that underlie its pathogen-blocking properties. In this work, we propose a new approach for studying the metabolic interactions between Aedes aegypti and Wolbachia using genome-scale models, finding that pathogen blocking is mainly influenced by competition for the resources required for Wolbachia and viral replication.}, } @article {pmid34620712, year = {2021}, author = {Xiao, Y and Chen, H and Wang, H and Zhang, M and Chen, X and Berk, JM and Zhang, L and Wei, Y and Li, W and Cui, W and Wang, F and Wang, Q and Cui, C and Li, T and Chen, C and Ye, S and Zhang, L and Ji, X and Huang, J and Wang, W and Wang, Z and Hochstrasser, M and Yang, H}, title = {Structural and mechanistic insights into the complexes formed by Wolbachia cytoplasmic incompatibility factors.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {118}, number = {41}, pages = {}, pmid = {34620712}, issn = {1091-6490}, support = {R01 GM046904/GM/NIGMS NIH HHS/United States ; R35 GM136325/GM/NIGMS NIH HHS/United States ; R37 GM046904/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified ; Drosophila melanogaster/genetics/*microbiology ; Embryo, Nonmammalian/*embryology ; Embryonic Development ; Female ; Infertility, Male/*physiopathology ; Male ; Mosquito Control/methods ; Multiprotein Complexes/metabolism ; Protein Binding ; Reproduction/*physiology ; Symbiosis ; Vector Borne Diseases/prevention & control/transmission/virology ; Wolbachia/*metabolism ; }, abstract = {Wolbachia bacteria, inherited through the female germ line, infect a large fraction of arthropod species. Many Wolbachia strains manipulate host reproduction, most commonly through cytoplasmic incompatibility (CI). CI, a conditional male sterility, results when Wolbachia-infected male insects mate with uninfected females; viability is restored if the female is similarly infected (called "rescue"). CI is used to help control mosquito-borne viruses such as dengue and Zika, but its mechanisms remain unknown. The coexpressed CI factors CifA and CifB form stable complexes in vitro, but the timing and function of this interaction in the insect are unresolved. CifA expression in the female germ line is sufficient for rescue. We report high-resolution structures of a CI-factor complex, CinA-CinB, which utilizes a unique binding mode between the CinA rescue factor and the CinB nuclease; the structures were validated by biochemical and yeast growth analyses. Importantly, transgenic expression in Drosophila of a nonbinding CinA mutant, designed based on the CinA-CinB structure, suggests CinA expressed in females must bind CinB imported by sperm in order to rescue embryonic viability. Binding between cognate factors is conserved in an enzymatically distinct CI system, CidA-CidB, suggesting universal features in Wolbachia CI induction and rescue.}, } @article {pmid34617112, year = {2021}, author = {Xiao, Z and Tao, X and Xu, X and Zhu, C and Nian, X and Han, D and Wang, D and He, Y}, title = {A Comparative Study on the Biological Characteristics of Parthenogenetic and Bisexual Restored Trichogramma pretiosum Lines.}, journal = {Journal of economic entomology}, volume = {114}, number = {6}, pages = {2355-2360}, doi = {10.1093/jee/toab194}, pmid = {34617112}, issn = {1938-291X}, mesh = {Animals ; Female ; Fertility ; Humans ; *Hymenoptera ; Reproduction ; *Sexual and Gender Minorities ; *Wasps ; *Wolbachia ; }, abstract = {In order to investigate the effect of Wolbachia on fitness of their hosts, the biological characteristics of a primarily parthenogenetic line and a bisexual restored line (obtained by antibiotic treatment) of Trichogramma pretiosum were compared in the laboratory. Results indicated that both the mean longevity and fecundity of parthenogenetic line (14.2 d and 165 eggs/female) were significantly higher than those in bisexual restored line (8.4 d and 124.2 eggs/female). Both lines of T. pretiosum had the highest daily fecundity (23.3 eggs/female for parthenogenetic line and 19.8 eggs/female for bisexual restored line) on the first day during their reproduction period, and their survival rate and daily fecundity decreased gradually with age; however, the survival rate and daily fecundity of the parthenogenetic line were always higher than those of bisexual restored line, during the entire experimental period. There was no significant difference in emergence rate and deformity rate between the two T. pretiosum lines. The life-table parameter results indicated that net reproduction rate (R0) and mean generation time (T) in parthenogenetic line (133 and 16.8) were significantly higher than those in bisexual restored line (61.7 and 15.5); intrinsic rate of natural increase (rm) and finite rate of increase (λ) in parthenogenetic line (0.29 and 1.34) were higher than those in bisexual restored line (0.26 and 1.3). Based on these results, it can be confirmed that the removal of Wolbachia in the parthenogenetic line had an adverse effect on fitness of T. pretiosum.}, } @article {pmid34616942, year = {2021}, author = {Bech, N and Beltran-Bech, S and Chupeau, C and Peccoud, J and Thierry, M and Raimond, R and Caubet, Y and Sicard, M and Grève, P}, title = {Experimental evidence of Wolbachia introgressive acquisition between terrestrial isopod subspecies.}, journal = {Current zoology}, volume = {67}, number = {4}, pages = {455-464}, pmid = {34616942}, issn = {1674-5507}, abstract = {Wolbachia are the most widespread endosymbiotic bacteria in animals. In many arthropod host species, they manipulate reproduction via several mechanisms that favor their maternal transmission to offspring. Among them, cytoplasmic incompatibility (CI) promotes the spread of the symbiont by specifically decreasing the fertility of crosses involving infected males and uninfected females, via embryo mortality. These differences in reproductive efficiency may select for the avoidance of incompatible mating, a process called reinforcement, and thus contribute to population divergence. In the terrestrial isopod Porcellio dilatatus, the Wolbachia wPet strain infecting the subspecies P. d. petiti induces unidirectional CI with uninfected individuals of the subspecies P. d. dilatatus. To study the consequences of CI on P. d. dilatatus and P. d. petiti hybridization, mitochondrial haplotypes and Wolbachia infection dynamics, we used population cages seeded with different proportions of the 2 subspecies in which we monitored these genetic parameters 5 and 7 years after the initial setup. Analysis of microsatellite markers allowed evaluating the degree of hybridization between individuals of the 2 subspecies. These markers revealed an increase in P. d. dilatatus nuclear genetic signature in all mixed cages, reflecting an asymmetry in hybridization. Hybridization led to the introgressive acquisition of Wolbachia and mitochondrial haplotype from P. d. petiti into nuclear genomes dominated by alleles of P. d. dilatatus. We discuss these results with regards to Wolbachia effects on their host (CI and putative fitness cost), and to a possible reinforcement that may have led to assortative mating, as possible factors contributing to the observed results.}, } @article {pmid34607949, year = {2021}, author = {Beebe, NW and Pagendam, D and Trewin, BJ and Boomer, A and Bradford, M and Ford, A and Liddington, C and Bondarenco, A and De Barro, PJ and Gilchrist, J and Paton, C and Staunton, KM and Johnson, B and Maynard, AJ and Devine, GJ and Hugo, LE and Rasic, G and Cook, H and Massaro, P and Snoad, N and Crawford, JE and White, BJ and Xi, Z and Ritchie, SA}, title = {Releasing incompatible males drives strong suppression across populations of wild and Wolbachia-carrying Aedes aegypti in Australia.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {118}, number = {41}, pages = {}, pmid = {34607949}, issn = {1091-6490}, mesh = {Aedes/*microbiology/physiology ; Animals ; Arbovirus Infections/*prevention & control/transmission ; Arboviruses ; Australia ; Biological Control Agents ; Female ; Humans ; *Infertility, Male ; Male ; Mosquito Control/*methods ; Mosquito Vectors/microbiology ; Queensland ; Wolbachia/*metabolism ; }, abstract = {Releasing sterile or incompatible male insects is a proven method of population management in agricultural systems with the potential to revolutionize mosquito control. Through a collaborative venture with the "Debug" Verily Life Sciences team, we assessed the incompatible insect technique (IIT) with the mosquito vector Aedes aegypti in northern Australia in a replicated treatment control field trial. Backcrossing a US strain of Ae. aegypti carrying Wolbachia wAlbB from Aedes albopictus with a local strain, we generated a wAlbB2-F4 strain incompatible with both the wild-type (no Wolbachia) and wMel-Wolbachia Ae. aegypti now extant in North Queensland. The wAlbB2-F4 strain was manually mass reared with males separated from females using Verily sex-sorting technologies to obtain no detectable female contamination in the field. With community consent, we delivered a total of three million IIT males into three isolated landscapes of over 200 houses each, releasing ∼50 males per house three times a week over 20 wk. Detecting initial overflooding ratios of between 5:1 and 10:1, strong population declines well beyond 80% were detected across all treatment landscapes when compared to controls. Monitoring through the following season to observe the ongoing effect saw one treatment landscape devoid of adult Ae. aegypti early in the season. A second landscape showed reduced adults, and the third recovered fully. These encouraging results in suppressing both wild-type and wMel-Ae. aegypti confirms the utility of bidirectional incompatibility in the field setting, show the IIT to be robust, and indicate that the removal of this arbovirus vector from human-occupied landscapes may be achievable.}, } @article {pmid34588907, year = {2021}, author = {Tyagi, K and Tyagi, I and Kumar, V}, title = {Insights into the gut bacterial communities of spider from wild with no evidence of phylosymbiosis.}, journal = {Saudi journal of biological sciences}, volume = {28}, number = {10}, pages = {5913-5924}, pmid = {34588907}, issn = {1319-562X}, abstract = {In the present study, an effort has been made to elucidate the gut bacterial diversity of twelve species of the family Araneidae under three subfamilies collected from 5 states of India along with their predicted metabolic role in functional metabolism. Further, we also compared the host species phylogeny based on partial cytochrome c oxidase subunit I (COI) sequences with the gut bacteria composition dendrogram to decipher the phylosymbiotic relationships. Analysis revealed the presence of 22 bacterial phyla, 145 families, and 364 genera in the gut, with Proteobacteria, Firmicutes, Actinobacteria, and Deinococcus-Thermus as the highest abundant phyla. Moreover, phylum Bacteriodetes was dominated only in Cyclosa mulmeinensis and Chlamydiae in Neoscona bengalensis. At the genus level, Bacillus, Acinetobacter, Cutibacterium, Pseudomonas, and Staphylococcus were the most dominant genera. Furthermore, the genus Prevotella was observed only in Cyclosa mulmeinensis, and endosymbiont Wolbachia only in Eriovixia laglaizei. The differential abundance analysis (DeSeq2) revealed the 19 significant ASVs represented by the genera like Acinetobacter, Vagoccoccus, Prevotella, Staphylococcus, Curvibacter, Corynebacterium, Paracoccus, Streptococcus, Microbacterium, and Pseudocitrobacter. The inter- and intra-subfamilies comparison based on diversity indices (alpha and beta diversity) revealed that the subfamily Araneinae have high richness and diversity than Argiopinae and Gasteracanthinae. The phylosymbiotic analysis revealed that there is no congruence between the gut bacteria composition dendrogram with their host phylogeny.}, } @article {pmid34584163, year = {2021}, author = {Conceição, CC and da Silva, JN and Arcanjo, A and Nogueira, CL and de Abreu, LA and de Oliveira, PL and Gondim, KC and Moraes, B and de Carvalho, SS and da Silva, RM and da Silva Vaz, I and Moreira, LA and Logullo, C}, title = {Aedes fluviatilis cell lines as new tools to study metabolic and immune interactions in mosquito-Wolbachia symbiosis.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {19202}, pmid = {34584163}, issn = {2045-2322}, mesh = {Aedes/immunology/metabolism/*microbiology ; Animals ; Cell Line ; Female ; Host Microbial Interactions/immunology ; *Immunity, Innate ; Symbiosis/immunology ; Wolbachia/*immunology ; }, abstract = {In the present work, we established two novel embryonic cell lines from the mosquito Aedes fluviatilis containing or not the naturally occurring symbiont bacteria Wolbachia, which were called wAflu1 and Aflu2, respectively. We also obtained wAflu1 without Wolbachia after tetracycline treatment, named wAflu1.tet. Morphofunctional characterization was performed to help elucidate the symbiont-host interaction in the context of energy metabolism regulation and molecular mechanisms of the immune responses involved. The presence of Wolbachia pipientis improves energy performance in A. fluviatilis cells; it affects the regulation of key energy sources such as lipids, proteins, and carbohydrates, making the distribution of actin more peripheral and with extensions that come into contact with neighboring cells. Additionally, innate immunity mechanisms were activated, showing that the wAflu1 and wAflu1.tet cells are responsive after the stimulus using Gram negative bacteria. Therefore, this work confirms the natural, mutually co-regulating symbiotic relationship between W. pipientis and A. fluviatilis, modulating the host metabolism and immune pathway activation. The results presented here add important resources to the current knowledge of Wolbachia-arthropod interactions.}, } @article {pmid34580706, year = {2021}, author = {Bubnell, JE and Fernandez-Begne, P and Ulbing, CKS and Aquadro, CF}, title = {Diverse wMel variants of Wolbachia pipientis differentially rescue fertility and cytological defects of the bag of marbles partial loss of function mutation in Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {11}, number = {12}, pages = {}, pmid = {34580706}, issn = {2160-1836}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Calcium Carbonate ; *Drosophila melanogaster/genetics ; Female ; Fertility/genetics ; Loss of Function Mutation ; Phylogeny ; *Wolbachia ; }, abstract = {In Drosophila melanogaster, the maternally inherited endosymbiont Wolbachia pipientis interacts with germline stem cell genes during oogenesis. One such gene, bag of marbles (bam) is the key switch for differentiation and also shows signals of adaptive evolution for protein diversification. These observations have led us to hypothesize that W. pipientis could be driving the adaptive evolution of bam for control of oogenesis. To test this hypothesis, we must understand the specificity of the genetic interaction between bam and W. pipientis. Previously, we documented that the W. pipientis variant, wMel, rescued the fertility of the bamBW hypomorphic mutant as a transheterozygote over a bam null. However, bamBW was generated more than 20 years ago in an uncontrolled genetic background and maintained over a balancer chromosome. Consequently, the chromosome carrying bamBW accumulated mutations that have prevented controlled experiments to further assess the interaction. Here, we used CRISPR/Cas9 to engineer the same single amino acid bam hypomorphic mutation (bamL255F) and a new bam null disruption mutation into the w1118 isogenic background. We assess the fertility of wildtype bam, bamL255F/bamnull hypomorphic, and bamL255F/bamL255F mutant females, each infected individually with 10 W. pipientis wMel variants representing three phylogenetic clades. Overall, we find that all of the W. pipientis variants tested here rescue bam hypomorphic fertility defects with wMelCS-like variants exhibiting the strongest rescue effects. In addition, these variants did not increase wildtype bam female fertility. Therefore, both bam and W. pipientis interact in genotype-specific ways to modulate female fertility, a critical fitness phenotype.}, } @article {pmid34579766, year = {2021}, author = {Lee, H and Seo, MG and Lee, SH and Oem, JK and Kim, SH and Jeong, H and Kim, Y and Jheong, WH and Kwon, OD and Kwak, D}, title = {Relationship among bats, parasitic bat flies, and associated pathogens in Korea.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {503}, pmid = {34579766}, issn = {1756-3305}, support = {NIER-2019-01-01-006//national institute of environmental research/ ; NRF-2016R1D1A1B02015366//National Research Foundation/ ; }, mesh = {Animals ; Bacteria/classification/*genetics/pathogenicity ; Chiroptera/*parasitology ; Diptera/anatomy & histology/classification/*microbiology/*parasitology ; Disease Reservoirs/microbiology/parasitology ; Disease Vectors ; Female ; Genetic Variation ; Male ; Parasites/classification/*genetics/pathogenicity ; Phylogeny ; Republic of Korea ; Sequence Analysis, DNA ; }, abstract = {BACKGROUND: Bats are hosts for many ectoparasites and act as reservoirs for several infectious agents, some of which exhibit zoonotic potential. Here, species of bats and bat flies were identified and screened for microorganisms that could be mediated by bat flies.

METHODS: Bat species were identified on the basis of their morphological characteristics. Bat flies associated with bat species were initially morphologically identified and further identified at the genus level by analyzing the cytochrome c oxidase subunit I gene. Different vector-borne pathogens and endosymbionts were screened using PCR to assess all possible relationships among bats, parasitic bat flies, and their associated organisms.

RESULTS: Seventy-four bat flies were collected from 198 bats; 66 of these belonged to Nycteribiidae and eight to Streblidae families. All Streblidae bat flies were hosted by Rhinolophus ferrumequinum, known as the most common Korean bat. Among the 74 tested bat flies, PCR and nucleotide sequencing data showed that 35 (47.3%) and 20 (27.0%) carried Wolbachia and Bartonella bacteria, respectively, whereas tests for Anaplasma, Borrelia, Hepatozoon, Babesia, Theileria, and Coxiella were negative. Phylogenetic analysis revealed that Wolbachia endosymbionts belonged to two different supergroups, A and F. One sequence of Bartonella was identical to that of Bartonella isolated from Taiwanese bats.

CONCLUSIONS: The vectorial role of bat flies should be checked by testing the same pathogen and bacterial organisms by collecting blood from host bats. This study is of great interest in the fields of disease ecology and public health owing to the bats' potential to transmit pathogens to humans and/or livestock.}, } @article {pmid34576710, year = {2021}, author = {Hoffmann, A and Müller, T and Fingerle, V and Noll, M}, title = {Presence of Human Pathogens of the Borrelia burgdorferi sensu lato Complex Shifts the Sequence Read Abundances of Tick Microbiomes in Two German Locations.}, journal = {Microorganisms}, volume = {9}, number = {9}, pages = {}, pmid = {34576710}, issn = {2076-2607}, support = {Kap. 1549 TG78//Bayerisches Staatsministerium für Bildung und Kultus, Wissenschaft und Kunst/ ; }, abstract = {The distribution of human Lyme borreliosis (LB) is assumed random in Germany, indicating that the human pathogenic species of the Borrelia burgdorferi sensu lato complex (Bb) are similarly distributed as part of the tick microbiome. The aim of this study was to differentiate if the presence of Bb occurs with a defined tick microbiome composition. Furthermore, the effect of location on tick microbiome composition was addressed for two German locations. Therefore, nucleic acid extracts from 82 Borrelia-positive and 118 Borrelia-negative Ixodes ricinus ticks sampled from human hosts in both districts were selected. Nucleic acid extracts were used for human pathogenic Bb species diagnostics based on qPCR and multilocus sequence typing (MLST) and bacterial 16S rRNA gene amplicon sequencing followed by network analyses. As a result, the presence of Bb shifted the sequence read abundances of Candidatus Midichloria, Rickettsia, Pseudomonas, Staphylococcus, and Candidatus Neoehrlichia and their topological roles in the tick microbiome. Moreover, the location was less important in the tick microbiome composition but shifted significantly sequence read abundances of Pseudomonas and Wolbachia as well as the topological role of microbial members. Since the presence of human pathogenic Bb species with other tick-associated pathogens varies regionally, we suggest that a bacterial 16S rRNA gene-based microbiome survey should be implemented in the routine diagnostics for both tick and host if human pathogenic species of Bb were detected. This diagnostic extension will help to optimize therapeutic approaches against Bb infection and co-occurring pathogens.}, } @article {pmid34568917, year = {2021}, author = {Ettinger, CL and Byrne, FJ and Collin, MA and Carter-House, D and Walling, LL and Atkinson, PW and Redak, RA and Stajich, JE}, title = {Improved draft reference genome for the Glassy-winged Sharpshooter (Homalodisca vitripennis), a vector for Pierce's disease.}, journal = {G3 (Bethesda, Md.)}, volume = {11}, number = {10}, pages = {}, pmid = {34568917}, issn = {2160-1836}, support = {S10 OD016290/OD/NIH HHS/United States ; }, mesh = {Animals ; *Genome, Insect ; *Hemiptera/genetics ; Metagenome ; Pilot Projects ; *Xylella ; }, abstract = {Homalodisca vitripennis (Hemiptera: Cicadellidae), known as the glassy-winged sharpshooter, is a xylem feeding leafhopper and an important agricultural pest as a vector of Xylella fastidiosa, which causes Pierce's disease in grapes and a variety of other scorch diseases. The current H. vitripennis reference genome from the Baylor College of Medicine's i5k pilot project is a 1.4-Gb assembly with 110,000 scaffolds, which still has significant gaps making identification of genes difficult. To improve on this effort, we used a combination of Oxford Nanopore long-read sequencing technology combined with Illumina sequencing reads to generate a better assembly and first-pass annotation of the whole genome sequence of a wild-caught Californian (Tulare County) individual of H. vitripennis. The improved reference genome assembly for H. vitripennis is 1.93-Gb in length (21,254 scaffolds, N50 = 650 Mb, BUSCO completeness = 94.3%), with 33.06% of the genome masked as repetitive. In total, 108,762 gene models were predicted including 98,296 protein-coding genes and 10,466 tRNA genes. As an additional community resource, we identified 27 orthologous candidate genes of interest for future experimental work including phenotypic marker genes like white. Furthermore, as part of the assembly process, we generated four endosymbiont metagenome-assembled genomes, including a high-quality near complete 1.7-Mb Wolbachia sp. genome (1 scaffold, CheckM completeness = 99.4%). The improved genome assembly and annotation for H. vitripennis, curated set of candidate genes, and endosymbiont MAGs will be invaluable resources for future research of H. vitripennis.}, } @article {pmid34567077, year = {2021}, author = {Cao, R and Ren, Q and Luo, J and Tian, Z and Liu, W and Zhao, B and Li, J and Diao, P and Tan, Y and Qiu, X and Zhang, G and Wang, Q and Guan, G and Luo, J and Yin, H and Liu, G}, title = {Analysis of Microorganism Diversity in Haemaphysalis longicornis From Shaanxi, China, Based on Metagenomic Sequencing.}, journal = {Frontiers in genetics}, volume = {12}, number = {}, pages = {723773}, pmid = {34567077}, issn = {1664-8021}, abstract = {Ticks are dangerous ectoparasites of humans and animals, as they are important disease vectors and serve as hosts for various microorganisms (including a variety of pathogenic microorganisms). Diverse microbial populations coexist within the tick body. Metagenomic next-generation sequencing (mNGS) has been suggested to be useful for rapidly and accurately obtaining microorganism abundance and diversity data. In this study, we performed mNGS to analyze the microbial diversity of Haemaphysalis longicornis from Baoji, Shaanxi, China, with the Illumina HiSeq platform. We identified 189 microbial genera (and 284 species) from ticks in the region; the identified taxa included Anaplasma spp., Rickettsia spp., Ehrlichia spp., and other important tick-borne pathogens at the genus level as well as symbiotic microorganisms such as Wolbachia spp., and Candidatus Entotheonella. The results of this study provide insights into possible tick-borne diseases and reveal new tick-borne pathogens in this region. Additionally, valuable information for the biological control of ticks is provided. In conclusion, this study provides reference data for guiding the development of prevention and control strategies targeting ticks and tick-borne diseases in the region, which can improve the effectiveness of tick and tick-borne disease control.}, } @article {pmid34566927, year = {2021}, author = {Calle-Tobón, A and Holguin-Rocha, AF and Moore, C and Rippee-Brooks, M and Rozo-Lopez, P and Harrod, J and Fatehi, S and Rua-Uribe, GL and Park, Y and Londoño-Rentería, B}, title = {Blood Meals With Active and Heat-Inactivated Serum Modifies the Gene Expression and Microbiome of Aedes albopictus.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {724345}, pmid = {34566927}, issn = {1664-302X}, support = {P20 GM103638/GM/NIGMS NIH HHS/United States ; }, abstract = {The Asian "tiger mosquito" Aedes albopictus is currently the most widely distributed disease-transmitting mosquito in the world. Its geographical expansion has also allowed the expansion of multiple arboviruses like dengue, Zika, and chikungunya, to higher latitudes. Due to the enormous risk to global public health caused by mosquitoes species vectors of human disease, and the challenges in slowing their expansion, it is necessary to develop new and environmentally friendly vector control strategies. Among these, host-associated microbiome-based strategies have emerged as promising options. In this study, we performed an RNA-seq analysis on dissected abdomens of Ae. albopictus females from Manhattan, KS, United States fed with sugar and human blood containing either normal or heat-inactivated serum, to evaluate the effect of heat inactivation on gene expression, the bacteriome transcripts and the RNA virome of this mosquito species. Our results showed at least 600 genes with modified expression profile when mosquitoes were fed with normal vs. heat-inactivated-containing blood. These genes were mainly involved in immunity, oxidative stress, lipid metabolism, and oogenesis. Also, we observed bacteriome changes with an increase in transcripts of Actinobacteria, Rhodospirillaceae, and Anaplasmataceae at 6 h post-feeding. We also found that feeding with normal blood seems to particularly influence Wolbachia metabolism, demonstrated by a significant increase in transcripts of this bacteria in mosquitoes fed with blood containing normal serum. However, no differences were observed in the virome core of this mosquito population. These results suggest that heat and further inactivation of complement proteins in human serum may have profound effect on mosquito and microbiome metabolism, which could influence interpretation of the pathogen-host interaction findings when using this type of reagents specially when measuring the effect of Wolbachia in vector competence.}, } @article {pmid34564245, year = {2021}, author = {Carpenter, M and Peng, L and Smith, AH and Joffe, J and O'Connor, M and Oliver, KM and Russell, JA}, title = {Frequent Drivers, Occasional Passengers: Signals of Symbiont-Driven Seasonal Adaptation and Hitchhiking in the Pea Aphid, Acyrthosiphon pisum.}, journal = {Insects}, volume = {12}, number = {9}, pages = {}, pmid = {34564245}, issn = {2075-4450}, support = {1050098, 1754597, 1050128, 1754302//National Science Foundation/ ; }, abstract = {Insects harbor a variety of maternally inherited bacterial symbionts. As such, variation in symbiont presence/absence, in the combinations of harbored symbionts, and in the genotypes of harbored symbiont species provide heritable genetic variation of potential use in the insects' adaptive repertoires. Understanding the natural importance of symbionts is challenging but studying their dynamics over time can help to elucidate the potential for such symbiont-driven insect adaptation. Toward this end, we studied the seasonal dynamics of six maternally transferred bacterial symbiont species in the multivoltine pea aphid (Acyrthosiphon pisum). Our sampling focused on six alfalfa fields in southeastern Pennsylvania, and spanned 14 timepoints within the 2012 growing season, in addition to two overwintering periods. To test and generate hypotheses on the natural relevance of these non-essential symbionts, we examined whether symbiont dynamics correlated with any of ten measured environmental variables from the 2012 growing season, including some of known importance in the lab. We found that five symbionts changed prevalence across one or both overwintering periods, and that the same five species underwent such frequency shifts across the 2012 growing season. Intriguingly, the frequencies of these dynamic symbionts showed robust correlations with a subset of our measured environmental variables. Several of these trends supported the natural relevance of lab-discovered symbiont roles, including anti-pathogen defense. For a seventh symbiont-Hamiltonella defensa-studied previously across the same study periods, we tested whether a reported correlation between prevalence and temperature stemmed not from thermally varying host-level fitness effects, but from selection on co-infecting symbionts or on aphid-encoded alleles associated with this bacterium. In general, such "hitchhiking" effects were not evident during times with strongly correlated Hamiltonella and temperature shifts. However, we did identify at least one time period in which Hamiltonella spread was likely driven by selection on a co-infecting symbiont-Rickettsiella viridis. Recognizing the broader potential for such hitchhiking, we explored selection on co-infecting symbionts as a possible driver behind the dynamics of the remaining six species. Out of twelve examined instances of symbiont dynamics unfolding across 2-week periods or overwintering spans, we found eight in which the focal symbiont underwent parallel frequency shifts under single infection and one or more co-infection contexts. This supported the idea that phenotypic variation created by the presence/absence of individual symbionts is a direct target for selection, and that symbiont effects can be robust under co-habitation with other symbionts. Contrastingly, in two cases, we found that selection may target phenotypes emerging from symbiont co-infections, with specific species combinations driving overall trends for the focal dynamic symbionts, without correlated change under single infection. Finally, in three cases-including the one described above for Hamiltonella-our data suggested that incidental co-infection with a (dis)favored symbiont could lead to large frequency shifts for "passenger" symbionts, conferring no apparent cost or benefit. Such hitchhiking has rarely been studied in heritable symbiont systems. We propose that it is more common than appreciated, given the widespread nature of maternally inherited bacteria, and the frequency of multi-species symbiotic communities across insects.}, } @article {pmid34564228, year = {2021}, author = {Xu, X and Ridland, PM and Umina, PA and Gill, A and Ross, PA and Pirtle, E and Hoffmann, AA}, title = {High Incidence of Related Wolbachia across Unrelated Leaf-Mining Diptera.}, journal = {Insects}, volume = {12}, number = {9}, pages = {}, pmid = {34564228}, issn = {2075-4450}, support = {MT16004//Hort Innovation/ ; }, abstract = {The maternally inherited endosymbiont, Wolbachia pipientis, plays an important role in the ecology and evolution of many of its hosts by affecting host reproduction and fitness. Here, we investigated 13 dipteran leaf-mining species to characterize Wolbachia infections and the potential for this endosymbiont in biocontrol. Wolbachia infections were present in 12 species, including 10 species where the Wolbachia infection was at or near fixation. A comparison of Wolbachia relatedness based on the wsp/MLST gene set showed that unrelated leaf-mining species often shared similar Wolbachia, suggesting common horizontal transfer. We established a colony of Liriomyza brassicae and found adult Wolbachia density was stable; although Wolbachia density differed between the sexes, with females having a 20-fold higher density than males. Wolbachia density increased during L. brassicae development, with higher densities in pupae than larvae. We removed Wolbachia using tetracycline and performed reciprocal crosses between Wolbachia-infected and uninfected individuals. Cured females crossed with infected males failed to produce offspring, indicating that Wolbachia induced complete cytoplasmic incompatibility in L. brassicae. The results highlight the potential of Wolbachia to suppress Liriomyza pests based on approaches such as the incompatible insect technique, where infected males are released into populations lacking Wolbachia or with a different incompatible infection.}, } @article {pmid34563127, year = {2021}, author = {Deng, J and Assandri, G and Chauhan, P and Futahashi, R and Galimberti, A and Hansson, B and Lancaster, LT and Takahashi, Y and Svensson, EI and Duplouy, A}, title = {Wolbachia-driven selective sweep in a range expanding insect species.}, journal = {BMC ecology and evolution}, volume = {21}, number = {1}, pages = {181}, pmid = {34563127}, issn = {2730-7182}, mesh = {Animals ; Cyprus ; *DNA, Mitochondrial/genetics ; Female ; Genetic Variation ; Odonata/*genetics/*microbiology ; Phylogeny ; *Wolbachia ; }, abstract = {BACKGROUND: Evolutionary processes can cause strong spatial genetic signatures, such as local loss of genetic diversity, or conflicting histories from mitochondrial versus nuclear markers. Investigating these genetic patterns is important, as they may reveal obscured processes and players. The maternally inherited bacterium Wolbachia is among the most widespread symbionts in insects. Wolbachia typically spreads within host species by conferring direct fitness benefits, and/or by manipulating its host reproduction to favour infected over uninfected females. Under sufficient selective advantage, the mitochondrial haplotype associated with the favoured maternally-inherited symbiotic strains will spread (i.e. hitchhike), resulting in low mitochondrial genetic variation across the host species range.

METHOD: The common bluetail damselfly (Ischnura elegans: van der Linden, 1820) has recently emerged as a model organism for genetics and genomic signatures of range expansion during climate change. Although there is accumulating data on the consequences of such expansion on the genetics of I. elegans, no study has screened for Wolbachia in the damselfly genus Ischnura. Here, we present the biogeographic variation in Wolbachia prevalence and penetrance across Europe and Japan (including samples from 17 populations), and from close relatives in the Mediterranean area (i.e. I. genei: Rambur, 1842; and I. saharensis: Aguesse, 1958).

RESULTS: Our data reveal (a) multiple Wolbachia-strains, (b) potential transfer of the symbiont through hybridization, (c) higher infection rates at higher latitudes, and (d) reduced mitochondrial diversity in the north-west populations, indicative of hitchhiking associated with the selective sweep of the most common strain. We found low mitochondrial haplotype diversity in the Wolbachia-infected north-western European populations (Sweden, Scotland, the Netherlands, Belgium, France and Italy) of I. elegans, and, conversely, higher mitochondrial diversity in populations with low penetrance of Wolbachia (Ukraine, Greece, Montenegro and Cyprus). The timing of the selective sweep associated with infected lineages was estimated between 20,000 and 44,000 years before present, which is consistent with the end of the last glacial period about 20,000 years.

CONCLUSIONS: Our findings provide an example of how endosymbiont infections can shape spatial variation in their host evolutionary genetics during postglacial expansion. These results also challenge population genetic studies that do not consider the prevalence of symbionts in many insects, which we show can impact geographic patterns of mitochondrial genetic diversity.}, } @article {pmid34555085, year = {2021}, author = {Lau, MJ and Hoffmann, AA and Endersby-Harshman, NM}, title = {A diagnostic primer pair to distinguish between wMel and wAlbB Wolbachia infections.}, journal = {PloS one}, volume = {16}, number = {9}, pages = {e0257781}, pmid = {34555085}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Anaplasmataceae Infections/*diagnosis/veterinary ; Animals ; DNA Primers/*genetics ; Early Diagnosis ; Female ; Sensitivity and Specificity ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Detection of the Wolbachia endosymbiont in Aedes aegypti mosquitoes through real-time polymerase chain reaction assays is widely used during and after Wolbachia releases in dengue reduction trials involving the wMel and wAlbB strains. Although several different primer pairs have been applied in current successful Wolbachia releases, they cannot be used in a single assay to distinguish between these strains. Here, we developed a new diagnostic primer pair, wMwA, which can detect the wMel or wAlbB infection in the same assay. We also tested current Wolbachia primers and show that there is variation in their performance when they are used to assess the relative density of Wolbachia. The new wMwA primers provide an accurate and efficient estimate of the presence and density of both Wolbachia infections, with practical implications for Wolbachia estimates in field collected Ae. aegypti where Wolbachia releases have taken place.}, } @article {pmid34548405, year = {2021}, author = {Beckmann, JF and Van Vaerenberghe, K and Akwa, DE and Cooper, BS}, title = {A single mutation weakens symbiont-induced reproductive manipulation through reductions in deubiquitylation efficiency.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {118}, number = {39}, pages = {}, pmid = {34548405}, issn = {1091-6490}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/microbiology/*pathology ; Deubiquitinating Enzymes/metabolism ; Drosophila melanogaster/genetics/*microbiology ; Embryo, Nonmammalian/metabolism/*microbiology ; Female ; Male ; *Mutation ; *Symbiosis ; *Ubiquitination ; Wolbachia/*physiology ; }, abstract = {Animals interact with microbes that affect their performance and fitness, including endosymbionts that reside inside their cells. Maternally transmitted Wolbachia bacteria are the most common known endosymbionts, in large part because of their manipulation of host reproduction. For example, many Wolbachia cause cytoplasmic incompatibility (CI) that reduces host embryonic viability when Wolbachia-modified sperm fertilize uninfected eggs. Operons termed cifs control CI, and a single factor (cifA) rescues it, providing Wolbachia-infected females a fitness advantage. Despite CI's prevalence in nature, theory indicates that natural selection does not act to maintain CI, which varies widely in strength. Here, we investigate the genetic and functional basis of CI-strength variation observed among sister Wolbachia that infect Drosophila melanogaster subgroup hosts. We cloned, Sanger sequenced, and expressed cif repertoires from weak CI-causing wYak in Drosophila yakuba, revealing mutations suspected to weaken CI relative to model wMel in D. melanogaster A single valine-to-leucine mutation within the deubiquitylating (DUB) domain of the wYak cifB homolog (cidB) ablates a CI-like phenotype in yeast. The same mutation reduces both DUB efficiency in vitro and transgenic CI strength in the fly, each by about twofold. Our results map hypomorphic transgenic CI to reduced DUB activity and indicate that deubiquitylation is central to CI induction in cid systems. We also characterize effects of other genetic variation distinguishing wMel-like cifs Importantly, CI strength determines Wolbachia prevalence in natural systems and directly influences the efficacy of Wolbachia biocontrol strategies in transinfected mosquito systems. These approaches rely on strong CI to reduce human disease.}, } @article {pmid34544126, year = {2021}, author = {Zhao, Z and Zhu, J and Hoffmann, AA and Cao, L and Shen, L and Fang, J and Ma, S and Liu, Q and Yu, W and Tang, L and Wang, Y and Jiang, W}, title = {Horizontal transmission and recombination of Wolbachia in the butterfly tribe Aeromachini Tutt, 1906 (Lepidoptera: Hesperiidae).}, journal = {G3 (Bethesda, Md.)}, volume = {11}, number = {9}, pages = {}, pmid = {34544126}, issn = {2160-1836}, mesh = {Animals ; *Butterflies/genetics ; Phylogeny ; Recombination, Genetic ; Species Specificity ; *Wolbachia/genetics ; }, abstract = {Wolbachia is arguably one of the most ubiquitous heritable symbionts among insects and understanding its transmission dynamics is crucial for understanding why it is so common. While previous research has studied the transmission pathways of Wolbachia in several insect lineages including Lepidoptera, this study takes advantage of data collected from the lepidopteran tribe Aeromachini in an effort to assess patterns of transmission. Twenty-one of the 46 species of Aeromachini species were infected with Wolbachia. Overall, 25% (31/125) of Aeromachini specimens tested were Wolbachia positive. All Wolbachia strains were species-specific except for the wJho strain which appeared to be shared by three host species with a sympatric distribution based on a cophylogenetic comparison between Wolbachia and the Aeromachini species. Two tests of phylogenetic congruence did not find any evidence for cospeciation between Wolbachia strains and their butterfly hosts. The cophylogenetic comparison, divergence time estimation, and Wolbachia recombination analysis revealed that Wolbachia acquisition in Aeromachini appears to have mainly occurred mainly through horizontal transmission rather than codivergence.}, } @article {pmid34533100, year = {2022}, author = {Shi, Y and Zheng, B}, title = {Discrete dynamical models on Wolbachia infection frequency in mosquito populations with biased release ratios.}, journal = {Journal of biological dynamics}, volume = {16}, number = {1}, pages = {320-339}, doi = {10.1080/17513758.2021.1977400}, pmid = {34533100}, issn = {1751-3766}, mesh = {*Aedes ; Animals ; Female ; Humans ; Male ; Models, Biological ; *Wolbachia ; }, abstract = {We develop two discrete models to study how supplemental releases affect the Wolbachia spreading dynamics in cage mosquito populations. The first model focuses on the case when only infected males are released at each generation. This release strategy has been proved to be capable of speeding up the Wolbachia persistence by suppressing the compatible matings between uninfected individuals. The second model targets the case when only infected females are released at each generation. For both models, detailed model formulation, enumeration of the positive equilibria and their stability analysis are provided. Theoretical results show that the two models can generate bistable dynamics when there are three positive equilibrium points, semi-stable dynamics for the case of two positive equilibrium points. And when the positive equilibrium point is unique, it is globally asymptotically stable. Some numerical simulations are offered to get helpful implications on the design of the release strategy.}, } @article {pmid34532954, year = {2022}, author = {Bagheri, Z and Talebi, AA and Asgari, S and Mehrabadi, M}, title = {Wolbachia promotes successful sex with siblings in the parasitoid Habrobracon hebetor.}, journal = {Pest management science}, volume = {78}, number = {1}, pages = {362-368}, doi = {10.1002/ps.6649}, pmid = {34532954}, issn = {1526-4998}, support = {//Tarbiat Modares University/ ; 98008582//Iran National Science Foundation/ ; }, mesh = {Animals ; Female ; Humans ; Inbreeding ; Male ; Reproduction ; Siblings ; *Wasps ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Wolbachia are intracellular α-proteobacteria that have a wide distribution among various arthropods and nematodes. They affect the host reproduction favoring their maternal transmission, which sets up a potential conflict in inbreeding situations when the host avoids sexual reproduction preventing inbreeding depression, while Wolbachia pushes it. We used the wasp Habrobracon hebetor to test the hypothesis that Wolbachia modulates inbreeding avoidance behavior and promotes sib mating.

RESULTS: Our results showed no obvious pre-copulatory inbreeding avoidance in this wasp. However, H. hebetor showed a strong post-copulatory inbreeding avoidance behavior that resulted in a low fertilization rate of uninfected siblings and therefore high rate of production of male progeny was obtained. We observed higher rates of fertilization success in the Wolbachia-infected lines that resulted in significantly higher female progeny production compared to the uninfected sib mates. Since diploid females are the result of successful fertilization due to haplodiploidy sex determination system in this insect, our results indicate that Wolbachia promoted fertile sib mating in H. hebetor. Interestingly, the rate of adult emergence in the progeny of Wolbachia-infected sib mates were almost similar to the non-sib mate crosses and significantly more than those observed in the uninfected sib mate crosses.

CONCLUSION: Our results support the idea that Wolbachia modulates inbreeding avoidance and promotes sib mating and also mitigates inbreeding depression. By promoting successful sex with siblings and increasing the probability of female progeny, Wolbachia enhances its transmission to the next generation. This is an undescribed effect of Wolbachia on the host reproduction. © 2021 Society of Chemical Industry.}, } @article {pmid34527601, year = {2021}, author = {Altinli, M and Schnettler, E and Sicard, M}, title = {Symbiotic Interactions Between Mosquitoes and Mosquito Viruses.}, journal = {Frontiers in cellular and infection microbiology}, volume = {11}, number = {}, pages = {694020}, pmid = {34527601}, issn = {2235-2988}, mesh = {Animals ; *Arboviruses ; *Culicidae ; Female ; Humans ; *Insect Viruses ; Metagenomics ; *Viruses ; }, abstract = {Mosquitoes not only transmit human and veterinary pathogens called arboviruses (arthropod-borne viruses) but also harbor mosquito-associated insect-specific viruses (mosquito viruses) that cannot infect vertebrates. In the past, studies investigating mosquito viruses mainly focused on highly pathogenic interactions that were easier to detect than those without visible symptoms. However, the recent advances in viral metagenomics have highlighted the abundance and diversity of viruses which do not generate mass mortality in host populations. Over the last decade, this has facilitated the rapid growth of virus discovery in mosquitoes. The circumstances around the discovery of mosquito viruses greatly affected how they have been studied so far. While earlier research mainly focused on the pathogenesis caused by DNA and some double-stranded RNA viruses during larval stages, more recently discovered single-stranded RNA mosquito viruses were heavily studied for their putative interference with arboviruses in female adults. Thus, many aspects of mosquito virus interactions with their hosts and host-microbiota are still unknown. In this context, considering mosquito viruses as endosymbionts can help to identify novel research areas, in particular in relation to their long-term interactions with their hosts (e.g. relationships during all life stages, the stability of the associations at evolutionary scales, transmission routes and virulence evolution) and the possible context-dependent range of interactions (i.e. beneficial to antagonistic). Here, we review the symbiotic interactions of mosquito viruses considering different aspects of their ecology, such as transmission, host specificity, host immune system and interactions with other symbionts within the host cellular arena. Finally, we highlight related research gaps in mosquito virus research.}, } @article {pmid34527179, year = {2021}, author = {McGillan, P and Berry, NG and Nixon, GL and Leung, SC and Webborn, PJH and Wenlock, MC and Kavanagh, S and Cassidy, A and Clare, RH and Cook, DA and Johnston, KL and Ford, L and Ward, SA and Taylor, MJ and Hong, WD and O'Neill, PM}, title = {Development of Pyrazolopyrimidine Anti-Wolbachia Agents for the Treatment of Filariasis.}, journal = {ACS medicinal chemistry letters}, volume = {12}, number = {9}, pages = {1421-1426}, pmid = {34527179}, issn = {1948-5875}, abstract = {Anti-Wolbachia therapy has been identified as a viable treatment for combating filarial diseases. Phenotypic screening revealed a series of pyrazolopyrimidine hits with potent anti-Wolbachia activity. This paper focuses on the exploration of the SAR for this chemotype, with improvement of metabolic stability and solubility profiles using medicinal chemistry approaches. Organic synthesis has enabled functionalization of the pyrazolopyrimidine core at multiple positions, generating a library of compounds of which many analogues possess nanomolar activity against Wolbachia in vitro with improved DMPK parameters. A lead compound, 15f, was selected for in vivo pharmacokinetics (PK) profiling in mice. The combination of potent anti-Wolbachia activity in two in vitro assessments plus the exceptional oral PK profiles in mice puts this lead compound in a strong position for in vivo proof-of-concept pharmacodynamics studies and demonstrates the strong potential for further optimization and development of this series for treatment of filariasis in the future.}, } @article {pmid34525260, year = {2022}, author = {Chen, F and Schenkel, M and Geuverink, E and van de Zande, L and Beukeboom, LW}, title = {Absence of complementary sex determination in two Leptopilina species (Figitidae, Hymenoptera) and a reconsideration of its incompatibility with endosymbiont-induced thelytoky.}, journal = {Insect science}, volume = {29}, number = {3}, pages = {900-914}, pmid = {34525260}, issn = {1744-7917}, support = {824.15.015//Netherlands Organisation for Scientific Research (NWO)/ ; 201506300038//China Scholarship Council/ ; }, mesh = {Animals ; Diploidy ; Female ; Haploidy ; *Hymenoptera/genetics/microbiology ; Male ; Parthenogenesis ; *Wasps/genetics/microbiology ; *Wolbachia/genetics ; }, abstract = {Complementary sex determination (CSD) is a widespread sex determination mechanism in haplodiploid Hymenoptera. Under CSD, sex is determined by the allelic state of one or multiple CSD loci. Heterozygosity at one or more loci leads to female development, whereas hemizygosity of haploid eggs and homozygosity of diploid eggs results in male development. Sexual (arrhenotokous) reproduction normally yields haploid male and diploid female offspring. Under asexual reproduction (thelytoky), diploidized unfertilized eggs develop into females. Thelytoky is often induced by bacterial endosymbionts that achieve egg diploidization by gamete duplication. As gamete duplication leads to complete homozygosity, endosymbiont-induced thelytokous reproduction is presumed to be incompatible with CSD, which relies on heterozygosity for female development. Previously, we excluded CSD in four Asobara (Braconidae) species and proposed a two-step mechanism for Wolbachia-induced thelytoky in Asobara japonica. Here, we conclusively reject CSD in two cynipid wasp species, Leptopilina heterotoma and Leptopilina clavipes. We further show that thelytoky in L. clavipes depends on Wolbachia titer but that diploidization and feminization steps cannot be separated, unlike in A. japonica. We discuss what these results reveal about the sex determination mechanism of L. clavipes and the presumed incompatibility between CSD and endosymbiont-induced thelytoky in the Hymenoptera.}, } @article {pmid34506623, year = {2021}, author = {Xu, S and Chen, J and Qin, M and Jiang, L and Qiao, G}, title = {Geography-dependent symbiont communities in two oligophagous aphid species.}, journal = {FEMS microbiology ecology}, volume = {97}, number = {10}, pages = {}, pmid = {34506623}, issn = {1574-6941}, mesh = {Animals ; *Aphids ; *Buchnera/genetics ; Geography ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; }, abstract = {Aphids and their diverse symbionts have become a good model to study bacteria-arthropod symbiosis. The feeding habits of aphids are usually influenced by a variety of symbionts. Most studies on symbiont diversity have focused on polyphagous aphids, while symbiont community patterns for oligophagous aphids remain unclear. Here, we surveyed the bacterial communities in natural populations of two oligophagous aphids, Melanaphis sacchari and Neophyllaphis podocarpi, in natural populations. Seven common symbionts were detected, among which Buchnera aphidicola and Wolbachia were the most prevalent. In addition, an uncommon Sodalis-like symbiont was also detected in these two aphids, and Gilliamella was found in some samples of M. sacchari. We further assessed the significant variation in symbiont communities within the two aphid species, geographical regions and host specialization using statistical and ordination analyses. Geography was an important factor in shaping the symbiont community structure in these oligophagous aphids. Furthermore, the strong geographical influence may be related to specific environmental factors, especially temperature, among different regions. These findings extend our knowledge of the significance of geography and its associated environmental conditions in the symbiont community structure associated with oligophagous aphids.}, } @article {pmid34495683, year = {2021}, author = {Li, TP and Zha, SS and Zhou, CY and Xia, X and Hoffmann, AA and Hong, XY}, title = {Two Newly Introduced Wolbachia Endosymbionts Induce Cell Host Differences in Competitiveness and Metabolic Responses.}, journal = {Applied and environmental microbiology}, volume = {87}, number = {22}, pages = {e0147921}, pmid = {34495683}, issn = {1098-5336}, support = {2016YFC1201200//MOST | National Key Research and Development Program of China (973 Program)/ ; 32020103011//National Natural Science Foundation of China (NSFC)/ ; }, mesh = {Aedes/microbiology ; Animals ; Cell Line ; Hemiptera/*microbiology ; *Symbiosis ; *Wolbachia/classification/growth & development ; }, abstract = {Wolbachia endosymbionts can induce multiple reproductive manipulations in their hosts, with cytoplasmic incompatibility (CI) being one of the most common manipulations. Two important agricultural pests, the white-backed planthopper (Sogatella furcifera) and the brown planthopper (Nilaparvata lugens), are usually infected with CI-inducing Wolbachia strain wFur and non-CI-inducing Wolbachia strain wLug, respectively. The biological effects of these infections when present in a host cell are unknown. Here, we introduced the two Wolbachia strains into an Aedes albopictus cell line to stably establish a wFur-infected cell line (WFI) and a wLug-infected cell line (WLI). In a mixed culture, WFI cells were completely replaced by WLI cells, pointing to a stronger competitiveness of the WLI cell line. We found that infection by both Wolbachia strains reduced cell growth rates, but WLI had a higher cell growth rate than WFI, and this difference in cell growth rate combined with possible Wolbachia differences in diffusivity may have affected cell competitiveness. By examining gene expression and metabolites in the two lines, we found that some genes and key metabolites responded to differences in cell competitiveness. These results point to potential mechanisms that could contribute to the relative performance of hosts infected by these strains and also highlight the substantial impact of a non-CI Wolbachia on metabolism, which may in turn influence the fitness of its native host. IMPORTANCEWolbachia transinfection in insects can be used to suppress pests and block virus transmission. We stably introduced two Wolbachia strains from rice planthoppers into cell lines of an important arbovirus mosquito vector, Aedes albopictus. The levels of competitiveness of host cells from the lines infected by the two Wolbachia strains were different, as were metabolic responses of the cell lines. These results suggest potential metabolic effects of Wolbachia on native hosts that could be exploited when they are transinfected into novel hosts for pest control.}, } @article {pmid34493670, year = {2021}, author = {Jing, YP and Wen, X and Li, L and Zhang, S and Zhang, C and Zhou, S}, title = {The vitellogenin receptor functionality of the migratory locust depends on its phosphorylation by juvenile hormone.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {118}, number = {37}, pages = {}, pmid = {34493670}, issn = {1091-6490}, mesh = {Animals ; Egg Proteins/*metabolism ; Endocytosis ; Female ; Isoenzymes/metabolism ; Juvenile Hormones/*pharmacology ; Locusta migratoria ; Oocytes/cytology/drug effects/*physiology ; Phosphorylation ; Protein Kinase C/metabolism ; Receptors, Cell Surface/*metabolism ; Receptors, G-Protein-Coupled/metabolism ; Type C Phospholipases/metabolism ; *Vitellogenesis ; Vitellogenins/*metabolism ; }, abstract = {Vitellogenin receptor (VgR) plays a pivotal role in ovarian vitellogenin (Vg) uptake and vertical transmission of pathogenic microbes and Wolbachia symbionts. However, the regulatory mechanisms of VgR action as an endocytic receptor and translocation from oocyte cytoplasm to the membrane remain poorly understood. Here, by using the migratory locust Locusta migratoria as a model system, we report that juvenile hormone (JH) promotes VgR phosphorylation at Ser[1361] in the second EGF-precursor homology domain. A signaling cascade including GPCR, PLC, extracellular calcium, and PKC-ι is involved in JH-stimulated VgR phosphorylation. This posttranslational regulation is a prerequisite for VgR binding to Vg on the external surface of the oocyte membrane and subsequent VgR/Vg endocytosis. Acidification, a condition in endosomes, induces VgR dephosphorylation along with the dissociation of Vg from VgR. Phosphorylation modification is also required for VgR recycling from oocyte cytoplasm to the membrane. Additionally, VgR phosphorylation and its requirement for Vg uptake and VgR recycling are evolutionarily conserved in other representative insects including the cockroach Periplaneta americana and the cotton bollworm Helicoverpa armigera This study fills an important knowledge gap of low-density lipoprotein receptors in posttranslational regulation, endocytosis, and intracellular recycling.}, } @article {pmid34492433, year = {2021}, author = {Parry, R and de Malmanche, H and Asgari, S}, title = {Persistent Spodoptera frugiperda rhabdovirus infection in Sf9 cells is not restricted by Wolbachia wMelPop-CLA and wAlbB strains and is targeted by the RNAi machinery.}, journal = {Virology}, volume = {563}, number = {}, pages = {82-87}, doi = {10.1016/j.virol.2021.08.013}, pmid = {34492433}, issn = {1096-0341}, mesh = {Animals ; Cell Line ; Genome, Viral ; Host-Pathogen Interactions ; RNA Interference ; RNA, Viral ; Rhabdoviridae/*physiology ; Spodoptera/*virology ; Wolbachia/classification/*physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia pipientis confers RNA virus refractoriness in Drosophila and Aedes mosquitoes. Questions remain about the Wolbachia-virus restriction phenotype and how extensive this phenomenon may be within other arthropods. Here, we generated two Spodoptera frugiperda cell lines stably transinfected with two strains of Wolbachia, wAlbB and wMelPop-CLA. Despite the high density of Wolbachia in stably infected Sf9 cells, RT-PCR indicated the presence of the negative-sense RNA virus Spodoptera frugiperda rhabdovirus (SfRV) in Wolbachia-infected and uninfected cell lines. No differences in the replication of SfRV between Sf9 and Wolbachia-infected cells was found. RNA-Seq analysis of the parental Sf9 cells supported SfRV's presence in these cells with abundant 20 nt virus-derived small RNAs indicating active replication of SfRV in these cells. Overall, this study supports a growing body of evidence that Wolbachia does not restrict negative-sense RNA viruses and generates an in vitro model to examine Lepidoptera-Wolbachia virus interactions.}, } @article {pmid34492017, year = {2021}, author = {Pocquet, N and O'Connor, O and Flores, HA and Tutagata, J and Pol, M and Hooker, DJ and Inizan, C and Russet, S and Duyvestyn, JM and Pacidônio, EC and Girault, D and da Silva Gonçalves, D and Minier, M and Touzain, F and Chalus, E and Lucien, K and Cheilan, F and Derycke, T and Laumond, S and Simmons, CP and Dupont-Rouzeyrol, M and Rossi, N}, title = {Assessment of fitness and vector competence of a New Caledonia wMel Aedes aegypti strain before field-release.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {9}, pages = {e0009752}, pmid = {34492017}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Chikungunya virus/physiology ; Dengue Virus/classification/physiology ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology ; New Caledonia ; Pest Control, Biological/*methods ; *Wolbachia ; Zika Virus/classification ; }, abstract = {BACKGROUND: Biological control programs involving Wolbachia-infected Aedes aegypti are currently deployed in different epidemiological settings. New Caledonia (NC) is an ideal location for the implementation and evaluation of such a strategy as the only proven vector for dengue virus (DENV) is Ae. aegypti and dengue outbreaks frequency and severity are increasing. We report the generation of a NC Wolbachia-infected Ae. aegypti strain and the results of experiments to assess the vector competence and fitness of this strain for future implementation as a disease control strategy in Noumea, NC.

METHODS/PRINCIPAL FINDINGS: The NC Wolbachia strain (NC-wMel) was obtained by backcrossing Australian AUS-wMel females with New Caledonian Wild-Type (NC-WT) males. Blocking of DENV, chikungunya (CHIKV), and Zika (ZIKV) viruses were evaluated via mosquito oral feeding experiments and intrathoracic DENV challenge. Significant reduction in infection rates were observed for NC-wMel Ae. aegypti compared to WT Ae. aegypti. No transmission was observed for NC-wMel Ae. aegypti. Maternal transmission, cytoplasmic incompatibility, fertility, fecundity, wing length, and insecticide resistance were also assessed in laboratory experiments. Ae. aegypti NC-wMel showed complete cytoplasmic incompatibility and a strong maternal transmission. Ae. aegypti NC-wMel fitness seemed to be reduced compared to NC-WT Ae. aegypti and AUS-wMel Ae. aegypti regarding fertility and fecundity. However further experiments are required to assess it accurately.

CONCLUSIONS/SIGNIFICANCE: Our results demonstrated that the NC-wMel Ae. aegypti strain is a strong inhibitor of DENV, CHIKV, and ZIKV infection and prevents transmission of infectious viral particles in mosquito saliva. Furthermore, our NC-wMel Ae. aegypti strain induces reproductive cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, supporting field-releases in Noumea, NC.}, } @article {pmid34488458, year = {2021}, author = {Chrostek, E and Martins, N and Marialva, MS and Teixeira, L}, title = {Wolbachia-Conferred Antiviral Protection Is Determined by Developmental Temperature.}, journal = {mBio}, volume = {12}, number = {5}, pages = {e0292320}, pmid = {34488458}, issn = {2150-7511}, support = {/WT_/Wellcome Trust/United Kingdom ; 094664/Z/10/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Dicistroviridae/*physiology ; Drosophila melanogaster/*growth & development/*microbiology/virology ; Female ; Host-Pathogen Interactions/*physiology ; Male ; *Temperature ; Viral Load ; Virus Diseases/prevention & control ; Virus Replication ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is a maternally transmitted bacterium that is widespread in arthropods and filarial nematodes and confers strong antiviral protection in Drosophila melanogaster and other arthropods. Wolbachia-transinfected Aedes aegypti mosquitoes are currently being deployed to fight transmission of dengue and Zika viruses. However, the mechanism of antiviral protection and the factors influencing are still not fully understood. Here, we show that temperature modulates Wolbachia-conferred protection in Drosophila melanogaster. Temperature after infection directly impacts Drosophila C virus (DCV) replication and modulates Wolbachia protection. At higher temperatures, viruses proliferate more and are more lethal, while Wolbachia confers lower protection. Strikingly, host developmental temperature is a determinant of Wolbachia-conferred antiviral protection. While there is strong protection when flies develop from egg to adult at 25°C, the protection is highly reduced or abolished when flies develop at 18°C. However, Wolbachia-induced changes during development are not sufficient to limit virus-induced mortality, as Wolbachia is still required to be present in adults at the time of infection. This developmental effect is general, since it was present in different host genotypes, Wolbachia variants, and upon infection with different viruses. Overall, we show that Wolbachia-conferred antiviral protection is temperature dependent, being present or absent depending on the environmental conditions. This interaction likely impacts Wolbachia-host interactions in nature and, as a result, frequencies of host and symbionts in different climates. Dependence of Wolbachia-mediated pathogen blocking on developmental temperature could be used to dissect the mechanistic bases of protection and influence the deployment of Wolbachia to prevent transmission of arboviruses. IMPORTANCE Insects are often infected with beneficial intracellular bacteria. The bacterium Wolbachia is extremely common in insects and can protect them from pathogenic viruses. This effect is being used to prevent transmission of dengue and Zika viruses by Wolbachia-infected mosquitoes. To understand the biology of insects in the wild, we need to discover which factors affect Wolbachia-conferred antiviral protection. Here, we show that the temperature at which insects develop from eggs to adults can determine the presence or absence of antiviral protection. The environment, therefore, strongly influences this insect-bacterium interaction. Our work may help to provide insights into the mechanism of viral blocking by Wolbachia, deepen our understanding of the geographical distribution of host and symbiont, and incentivize further research on the temperature dependence of Wolbachia-conferred protection for control of mosquito-borne disease.}, } @article {pmid34475664, year = {2021}, author = {Alipour, H and Izadpanah, L and Azizi, K and Shahriari-Namadi, M and Kalantari, M}, title = {Potential co-infection of Wolbachia with Leishmania among sand fly vectors caught from endemic leishmaniasis foci in Fars province, southern Iran.}, journal = {Journal of parasitic diseases : official organ of the Indian Society for Parasitology}, volume = {45}, number = {3}, pages = {817-822}, pmid = {34475664}, issn = {0971-7196}, abstract = {Leishmaniasis is one of the Neglected Tropical Diseases in the tropical region of many countries in the world. The etiological agents (Leishmania parasites) of the disease are transmitted to human and other vertebrate hosts by infectious bites of female phlebotomine sand flies. On the other hand, some symbiotic microorganisms such as Wolbachia (Rickettsiales: Anaplasmataceae) may be transmitted vertically in many arthropods and may cause synergistic or antagonistic effects on epidemiology of the vector-borne diseases. Hence, in the present study, potential coinfection of Wolbachia with Leishmania in the sand fly vectors will be examined by PCR technique in the important leishmaniasis foci of Fars province in southern Iran, as a new feature for the disease long-term control. Sand flies were collected by sticky traps from indoor and outdoor locations of 5 different areas of Fars province during 2018 and 2019. DNAs of sand flies were extracted and PCR method was performed based on primers which were designed from surface proteins (WSP) genome region for Wolbachia and minicircle kDNA gene for Leishmania detections. At last, PCR products were sequenced and recorded in the GenBank. Out of 1002 sand flies caught from 5 different foci of Fars province, 909 male and female and 386 female sand flies' DNAs were extracted for detection of Wolbachia and Leishmania by PCR, respectively. Accordingly, out of the total 44 pools prepared from sand flies, 6 out of 28 pools form P. papatasi female were positive for Wolbachia in Shiraz, Marvdasht and Kharameh. However, none of female sand flies were positive for Leishmania. The study also focused on monitoring of sand flies co-infection of Wolbachia with Leishmania, which was not found in any of the studied samples. The negative results may be due to control strategies implemented which were done against Leishmaniasis in the studied areas during last years.}, } @article {pmid34470970, year = {2022}, author = {Sasaki, T and Moi, ML and Saito, K and Isawa, H and Takasaki, T and Sawabe, K}, title = {Aedes albopictus Strain and Dengue Virus Serotype in the Dengue Fever Outbreaks in Japan: Implications of Wolbachia Infection.}, journal = {Japanese journal of infectious diseases}, volume = {75}, number = {2}, pages = {140-143}, doi = {10.7883/yoken.JJID.2021.376}, pmid = {34470970}, issn = {1884-2836}, mesh = {*Aedes/genetics/virology ; Anaplasmataceae Infections/microbiology/virology ; Animals ; *Dengue/epidemiology/immunology/virology ; *Dengue Virus/genetics/immunology ; *Disease Outbreaks ; Disease Susceptibility ; Japan/epidemiology ; Serogroup ; Symbiosis ; *Wolbachia/genetics/virology ; }, abstract = {From August 27 to October 15, 2014, a dengue fever outbreak with 158 autochthonous cases occurred after nearly 70 years of no reports of autochthonous cases in Japan. The most competent mosquito vector for dengue virus (DENV) transmission in Japan is Aedes albopictus. Since A. albopictus is widely distributed throughout Japan, we examined the susceptibility of this species to infection by DENV and the relationship of the endosymbiont Wolbachia (wAlbA and wAlbB) with susceptibility to DENV. The A. albopictus YYG strain, collected from the Yoyogi Park in 2014, the epicenter of the dengue fever outbreak, was found to have lower susceptibility to DENV 1 and 3 than that of the indigenous Japanese strains A. albopictus EBN 201808 (F1 from the field) and A. albopictus ISG 201603. Furthermore, the A. albopictus EBN 201808 strain showed the same susceptibility to DENV3 as the A. albopictus ISG 201603tet strain (Wolbachia-free). Susceptibility to DENV3 was not related to Wolbachia strains wAlbA or wAlbB in the A. albopictus ISG 201603 strain.}, } @article {pmid34468309, year = {2021}, author = {Dainty, KR and Hawkey, J and Judd, LM and Pacidônio, EC and Duyvestyn, JM and Gonçalves, DS and Lin, SY and O'Donnell, TB and O'Neill, SL and Simmons, CP and Holt, KE and Flores, HA}, title = {wMel Wolbachia genome remains stable after 7 years in Australian Aedes aegypti field populations.}, journal = {Microbial genomics}, volume = {7}, number = {9}, pages = {}, pmid = {34468309}, issn = {2057-5858}, support = {/WT_/Wellcome Trust/United Kingdom ; 212914/Z/18/B/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/virology ; Animals ; Australia ; Dengue ; Dengue Virus ; Evolution, Molecular ; *Genome, Bacterial ; Wolbachia/*genetics ; Zika Virus ; Zika Virus Infection ; }, abstract = {Infection of wMel Wolbachia in Aedes aegypti imparts two signature features that enable its application for biocontrol of dengue. First, the susceptibility of mosquitoes to viruses such as dengue and Zika is reduced. Second, a reproductive manipulation is caused that enables wMel introgression into wild-type mosquito populations. The long-term success of this method relies, in part, on evolution of the wMel genome not compromising the critical features that make it an attractive biocontrol tool. This study compared the wMel Wolbachia genome at the time of initial releases and 1-7 years post-release in Cairns, Australia. Our results show the wMel genome remains highly conserved up to 7 years post-release in gene sequence, content, synteny and structure. This work suggests the wMel genome is stable in its new mosquito host and, therefore, provides reassurance on the potential for wMel to deliver long-term public-health impacts.}, } @article {pmid34466651, year = {2021}, author = {Pupić-Bakrač, A and Pupić-Bakrač, J and Beck, A and Jurković, D and Polkinghorne, A and Beck, R}, title = {Dirofilaria repens microfilaremia in humans: Case description and literature review.}, journal = {One health (Amsterdam, Netherlands)}, volume = {13}, number = {}, pages = {100306}, pmid = {34466651}, issn = {2352-7714}, abstract = {INTRODUCTION: Dirofilaria repens is a vector-borne filaroid helminth of carnivorous animals, primarily domesticated dogs. Humans are considered to be accidental hosts in which D. repens rarely reach sexual maturity but induce local inflammation, mainly in subcutaneous and ocular tissues.

METHODS: In the current study, we present the detection of multiple adults of D. repens, endosymbiont Wolbachia sp. and microfilariae by molecular analysis in peripheral tissues and bloodstream of a human host. A subsequent meta-analysis of published literature identified 21 cases of human infection with adult D. repens producing microfilariae.

RESULTS: Within the study population, there were 13 (59.09%) males, eight (36.36%) females and, in one (4.55%) case, sex was not reported. A total of 11 (50.00%) cases had subcutaneous dirofilariasis, six (27.27%) had ocular dirofiliariasis, with single cases (4.55% each) of genital, mammary, lymphatic and a combination of subcutaneous and pulmonary dirofilariasis described. In one (4.55%) case, the primary anatomical site of adult D. repens could not be found. D. repens microfilariae were detected in the local tissue (local microfilariasis) in 11 (50.00%) cases and the peripheral blood (microfilaremia) in 11 (50.50%) cases. Final identification of D. repens microfilariae was based on morphological detection in 14 (63.64%) cases, and molecular detection in eight (36.36%) cases.

CONCLUSION: The results of this study suggest that humans may act as a final host for D. repens, however its role as a source of D. repens infection is less clear.}, } @article {pmid34455644, year = {2022}, author = {Bruzzese, DJ and Schuler, H and Wolfe, TM and Glover, MM and Mastroni, JV and Doellman, MM and Tait, C and Yee, WL and Rull, J and Aluja, M and Hood, GR and Goughnour, RB and Stauffer, C and Nosil, P and Feder, JL}, title = {Testing the potential contribution of Wolbachia to speciation when cytoplasmic incompatibility becomes associated with host-related reproductive isolation.}, journal = {Molecular ecology}, volume = {31}, number = {10}, pages = {2935-2950}, pmid = {34455644}, issn = {1365-294X}, support = {J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 31441/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Cytoplasm/genetics ; DNA, Mitochondrial/genetics ; Drosophila/genetics ; Male ; Reproductive Isolation ; *Tephritidae/genetics ; *Wolbachia/genetics ; }, abstract = {Endosymbiont-induced cytoplasmic incompatibility (CI) may play an important role in arthropod speciation. However, whether CI consistently becomes associated or coupled with other host-related forms of reproductive isolation (RI) to impede the transfer of endosymbionts between hybridizing populations and further the divergence process remains an open question. Here, we show that varying degrees of pre- and postmating RI exist among allopatric populations of two interbreeding cherry-infesting tephritid fruit flies (Rhagoletis cingulata and R. indifferens) across North America. These flies display allochronic and sexual isolation among populations, as well as unidirectional reductions in egg hatch in hybrid crosses involving southwestern USA males. All populations are infected by a Wolbachia strain, wCin2, whereas a second strain, wCin3, only co-infects flies from the southwest USA and Mexico. Strain wCin3 is associated with a unique mitochondrial DNA haplotype and unidirectional postmating RI, implicating the strain as the cause of CI. When coupled with nonendosymbiont RI barriers, we estimate the strength of CI associated with wCin3 would not prevent the strain from introgressing from infected southwestern to uninfected populations elsewhere in the USA if populations were to come into secondary contact and hybridize. In contrast, cytoplasmic-nuclear coupling may impede the transfer of wCin3 if Mexican and USA populations were to come into contact. We discuss our results in the context of the general paucity of examples demonstrating stable Wolbachia hybrid zones and whether the spread of Wolbachia among taxa can be constrained in natural hybrid zones long enough for the endosymbiont to participate in speciation.}, } @article {pmid34452330, year = {2021}, author = {Bhattacharya, T and Rice, DW and Crawford, JM and Hardy, RW and Newton, ILG}, title = {Evidence of Adaptive Evolution in Wolbachia-Regulated Gene DNMT2 and Its Role in the Dipteran Immune Response and Pathogen Blocking.}, journal = {Viruses}, volume = {13}, number = {8}, pages = {}, pmid = {34452330}, issn = {1999-4915}, support = {R21 AI137918/AI/NIAID NIH HHS/United States ; R21 AI153785/AI/NIAID NIH HHS/United States ; R01 AI144430/AI/NIAID NIH HHS/United States ; }, mesh = {Adaptation, Biological ; Aedes/enzymology/genetics/immunology/microbiology ; Amino Acid Sequence ; Animals ; DNA (Cytosine-5-)-Methyltransferases/chemistry/*genetics/immunology ; Diptera/classification/enzymology/*genetics/immunology ; Drosophila Proteins/chemistry/*genetics/immunology ; Drosophila melanogaster/*enzymology/genetics/immunology/*microbiology ; Evolution, Molecular ; *Host-Pathogen Interactions ; Phylogeny ; Protein Conformation ; Sequence Alignment ; Wolbachia/genetics/*physiology ; }, abstract = {Eukaryotic nucleic acid methyltransferase (MTase) proteins are essential mediators of epigenetic and epitranscriptomic regulation. DNMT2 belongs to a large, conserved family of DNA MTases found in many organisms, including holometabolous insects such as fruit flies and mosquitoes, where it is the lone MTase. Interestingly, despite its nomenclature, DNMT2 is not a DNA MTase, but instead targets and methylates RNA species. A growing body of literature suggests that DNMT2 mediates the host immune response against a wide range of pathogens, including RNA viruses. Curiously, although DNMT2 is antiviral in Drosophila, its expression promotes virus replication in mosquito species. We, therefore, sought to understand the divergent regulation, function, and evolution of these orthologs. We describe the role of the Drosophila-specific host protein IPOD in regulating the expression and function of fruit fly DNMT2. Heterologous expression of these orthologs suggests that DNMT2's role as an antiviral is host-dependent, indicating a requirement for additional host-specific factors. Finally, we identify and describe potential evidence of positive selection at different times throughout DNMT2 evolution within dipteran insects. We identify specific codons within each ortholog that are under positive selection and find that they are restricted to four distinct protein domains, which likely influence substrate binding, target recognition, and adaptation of unique intermolecular interactions. Collectively, our findings highlight the evolution of DNMT2 in Dipteran insects and point to structural, regulatory, and functional differences between mosquito and fruit fly homologs.}, } @article {pmid34448854, year = {2021}, author = {Hubert, J and Nesvorna, M and Pekar, S and Green, SJ and Klimov, PB}, title = {Cardinium inhibits Wolbachia in its mite host, Tyrophagus putrescentiae, and affects host fitness.}, journal = {FEMS microbiology ecology}, volume = {97}, number = {10}, pages = {}, doi = {10.1093/femsec/fiab123}, pmid = {34448854}, issn = {1574-6941}, mesh = {*Acaridae ; Animals ; Bacteroidetes/genetics ; Humans ; *Microbiota ; *Mites ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Interactions among endosymbiotic bacteria inside their eukaryotic hosts are poorly understood, particularly in mites. The mite Tyrophagus putrescentiae is a common, medically important generalist species that has many intracellular and gut bacterial symbionts. In the experiments, we examined bacterial abundances and composition in mite populations obtained by controlled mixing of stock mite populations that differed in the presence/absence of the major intracellular bacteria Wolbachia and Cardinium. Changes in microbial communities were characterized using 16S ribosomal RNA high-throughput sequencing (pooled mite individuals) and quantitative PCR for key microbial taxa (individual mites). Mite fitness was estimated as a parameter of population growth. We detected that in mixed mite populations, Cardinium and Wolbachia can co-occur in the same mite individual. The presence of Cardinium was negatively correlated with the presence of Wolbachia and Bartonella, while the Bartonella and Wolbachia were positively correlated in individual level samples. Since mixed populations had lower abundances of Wolbachia, while the abundance of Cardinium did not change, we suggest that the presence of Cardinium inhibits the growth of Wolbachia. The mixed mite populations had lower population growth than parental populations. The possible effect of symbionts on the fitness of mixed population is discussed.}, } @article {pmid34448004, year = {2021}, author = {Cruz, LNPD and Carvalho-Costa, LF and Rebêlo, JMM}, title = {Molecular Evidence Suggests That Wolbachia pipientis (Rickettsiales: Anaplasmataceae) is Widely Associated With South American Sand Flies (Diptera: Psychodidae).}, journal = {Journal of medical entomology}, volume = {58}, number = {6}, pages = {2186-2195}, doi = {10.1093/jme/tjab130}, pmid = {34448004}, issn = {1938-2928}, mesh = {Animals ; Brazil ; DNA, Bacterial/analysis ; Psychodidae/*physiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia pipientis (Hertig) is an endosymbiotic microorganism widespread among arthropods and other invertebrate hosts, and employed in strategies to reduce the incidence of arthropod-borne diseases. Here, we used a PCR-based approach for 16S RNA and wsp genes to investigate the prevalence, geographical distribution, and strains of Wolbachia in sand flies (Diptera: Psychodidae: Phlebotominae), the main vectors of the causative agents of leishmaniasis, from three biomes in Brazil: Amazon, Cerrado, and Caatinga. We found that: 1) Wolbachia DNA is present in most (66.7%) of the sampled sand fly species, including vectors of Leishmania spp. (Ross, Trypanosomatida: Trypanosomatidae), 2) the prevalence of Wolbachia DNA varies among species and populations, 3) some strains of Wolbachia may have wider geographical and host range in South America, and 4) two phylogenetic distinct wsp sequences might represent two novel strains for Wolbachia in South America sand flies. Those findings increase the basic knowledge about Wolbachia in South American sand flies and might foster further researches on its use to reduce the transmission of sand fly-borne parasites.}, } @article {pmid34446060, year = {2021}, author = {Bergman, A and Hesson, JC}, title = {Wolbachia prevalence in the vector species Culex pipiens and Culex torrentium in a Sindbis virus-endemic region of Sweden.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {428}, pmid = {34446060}, issn = {1756-3305}, support = {874735//Horizon 2020/ ; }, mesh = {Alphavirus Infections/*epidemiology/virology ; Animals ; Culex/classification/*microbiology ; Endemic Diseases ; Mosquito Vectors/*microbiology ; Prevalence ; Sindbis Virus/physiology ; Sweden/epidemiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia pipientis are endosymbiotic bacteria present in a large proportion of terrestrial arthropods. The species is known to sometimes affect the ability of its host to transmit vector-borne pathogens. Central Sweden is endemic for Sindbis virus (SINV), where it is mainly transmitted by the vector species Culex pipiens and Culex torrentium, with the latter established as the main vector. In this study we investigated the Wolbachia prevalence in these two vector species in a region highly endemic for SINV.

METHODS: Culex mosquitoes were collected using CDC light traps baited with carbon dioxide over 9 years at 50 collection sites across the River Dalälven floodplains in central Sweden. Mosquito genus was determined morphologically, while a molecular method was used for reliable species determination. The presence of Wolbachia was determined through PCR using general primers targeting the wsp gene and sequencing of selected samples.

RESULTS: In total, 676 Cx. pipiens and 293 Cx. torrentium were tested for Wolbachia. The prevalence of Wolbachia in Cx. pipiens was 97% (95% CI 94.8-97.6%), while only 0.7% (95% CI 0.19-2.45%) in Cx. torrentium. The two Cx. torrentium mosquitoes that were infected with Wolbachia carried different types of the bacteria.

CONCLUSIONS: The main vector of SINV in the investigated endemic region, Cx. torrentium, was seldom infected with Wolbachia, while it was highly prevalent in the secondary vector, Cx. pipiens. The presence of Wolbachia could potentially have an impact on the vector competence of these two species. Furthermore, the detection of Wolbachia in Cx. torrentium could indicate horizontal transmission of the endosymbiont between arthropods of different species.}, } @article {pmid34442732, year = {2021}, author = {Parry, R and James, ME and Asgari, S}, title = {Uncovering the Worldwide Diversity and Evolution of the Virome of the Mosquitoes Aedes aegypti and Aedes albopictus.}, journal = {Microorganisms}, volume = {9}, number = {8}, pages = {}, pmid = {34442732}, issn = {2076-2607}, support = {DP190102048//Australian Research Council/ ; }, abstract = {Aedes aegypti, the yellow fever mosquito, and Aedes albopictus, the Asian tiger mosquito, are the most significant vectors of dengue, Zika, and Chikungunya viruses globally. Studies examining host factors that control arbovirus transmission demonstrate that insect-specific viruses (ISVs) can modulate mosquitoes' susceptibility to arbovirus infection in both in vivo and in vitro co-infection models. While research is ongoing to implicate individual ISVs as proviral or antiviral factors, we have a limited understanding of the composition and diversity of the Aedes virome. To address this gap, we used a meta-analysis approach to uncover virome diversity by analysing ~3000 available RNA sequencing libraries representing a worldwide geographic range for both mosquitoes. We identified ten novel viruses and previously characterised viruses, including mononegaviruses, orthomyxoviruses, negeviruses, and a novel bi-segmented negev-like group. Phylogenetic analysis suggests close relatedness to mosquito viruses implying likely insect host range except for one arbovirus, the multi-segmented Jingmen tick virus (Flaviviridae) in an Italian colony of Ae. albopictus. Individual mosquito transcriptomes revealed remarkable inter-host variation of ISVs within individuals from the same colony and heterogeneity between different laboratory strains. Additionally, we identified striking virus diversity in Wolbachia infected Aedes cell lines. This study expands our understanding of the virome of these important vectors. It provides a resource for further assessing the ecology, evolution, and interaction of ISVs with their mosquito hosts and the arboviruses they transmit.}, } @article {pmid34442279, year = {2021}, author = {Su, CY and Zhu, DH and Yang, XH}, title = {Design and Testing of Effective Primers for Amplification of the orf7 Gene of Phage WO Associated with Andricus hakonensis.}, journal = {Insects}, volume = {12}, number = {8}, pages = {}, pmid = {34442279}, issn = {2075-4450}, support = {2016YFE0128200//National Key Research and Development Program of China/ ; }, abstract = {Phage WO was first characterized in Wolbachia, an obligate intracellular Rickettsiales known for its ability to regulate the reproduction of arthropod hosts. In this paper, we focus on the study of virus diversity in Andricus hakonensis and the development of highly effective primers. Based on the existing Wolbachia genome sequence, we designed primers (WO-TF and WO-TR) to amplify the full-length orf7 gene of phage WO. Surprisingly, sequencing results showed a high abundance of other phage WO groups in A. hakonensis, in addition to the four groups previously identified. The results also showed that A. hakonensis contained most of the known types of orf7 genes (I, III, IV, V and VI) and the level of diversity of harbored phage WO was very high. Therefore, we speculated that existing primers were not specific enough and that new primers for the detection of phage WO were needed. Based on the existing orf7 gene sequence, we designed specific detection primers (WO-SUF and WO-SUR). Sequencing results showed that the primers effectively amplified all known types of phage WO. In addition to amplifying most of the known sequences, we also detected some new genotypes in A. hakonensis using the new primers. Importantly, all phage WO groups could be efficiently detected. Combined with the results of previous studies, our results suggest that A. hakonensis contains the largest number of phage types (up to 36 types). This study is novel in that it provides practical molecular evidence supporting base deletions, in addition to gene mutations and genetic recombination, as an important cause of phage WO diversity.}, } @article {pmid34442272, year = {2021}, author = {Fallon, AM}, title = {Growth and Maintenance of Wolbachia in Insect Cell Lines.}, journal = {Insects}, volume = {12}, number = {8}, pages = {}, pmid = {34442272}, issn = {2075-4450}, abstract = {The obligate intracellular microbe, Wolbachia pipientis (Rickettsiales; Anaplasmataceae), is a Gram-negative member of the alpha proteobacteria that infects arthropods and filarial worms. Although closely related to the genera Anaplasma and Ehrlichia, which include pathogens of humans, Wolbachia is uniquely associated with invertebrate hosts in the clade Ecdysozoa. Originally described in Culex pipiens mosquitoes, Wolbachia is currently represented by 17 supergroups and is believed to occur in half of all insect species. In mosquitoes, Wolbachia acts as a gene drive agent, with the potential to modify vector populations; in filarial worms, Wolbachia functions as a symbiont, and is a target for drug therapy. A small number of Wolbachia strains from supergroups A, B, and F have been maintained in insect cell lines, which are thought to provide a more permissive environment than the natural host. When transferred back to an insect host, Wolbachia produced in cultured cells are infectious and retain reproductive phenotypes. Here, I review applications of insect cell lines in Wolbachia research and describe conditions that facilitate Wolbachia infection and replication in naive host cells. Progress in manipulation of Wolbachia in vitro will enable genetic and biochemical advances that will facilitate eventual genetic engineering of this important biological control agent.}, } @article {pmid34438106, year = {2021}, author = {Erban, T and Klimov, PB and Harant, K and Talacko, P and Nesvorna, M and Hubert, J}, title = {Label-free proteomic analysis reveals differentially expressed Wolbachia proteins in Tyrophagus putrescentiae: Mite allergens and markers reflecting population-related proteome differences.}, journal = {Journal of proteomics}, volume = {249}, number = {}, pages = {104356}, doi = {10.1016/j.jprot.2021.104356}, pmid = {34438106}, issn = {1876-7737}, mesh = {*Allergens ; Animals ; *Mites/microbiology ; Proteome ; Proteomics ; *Wolbachia ; }, abstract = {Tyrophagus putrescentiae is an astigmatid mite of great economic, medical and veterinary importance. The microbiome, especially intracellular bacteria, may affect allergy/allergen expression. We targeted Wolbachia proteins, allergen comparisons and markers in Wolbachia-mite interactions in three mite populations. A decoy database was constructed by proteogenomics using the T. putrescentiae draft genome, Wolbachia transcriptome assembly and current T. putrescentiae-related sequences in GenBank. Among thousands of mite-derived proteins, 18 Wolbachia proteins were reliably identified. We suggest that peroxiredoxin, bacterioferritin, ankyrin repeat domain-containing protein and DegQ family serine endoprotease indicate a higher-level bacterium-bacterium-host interaction. We produced evidence that the host-Wolbachia interaction is modulated through pattern recognition receptors (PRRs), mannose-binding lectins/mannose receptors, the cholinergic anti-inflammatory pathway with TNF-α, and others. We observed Tyr p 3 suppression in mites with Wolbachia, linking trypsin to PRR modulation. Nine out of the 12 current WHO/IUIS official allergens were reliably identified, but the remaining three allergens, Tyr p 1, 8 and 35, were detected as only trace hits. This study provides numerous markers for further Wolbachia-host interaction research. For accuracy, mite allergens should be considered according to abundance in species, but mite populations/strains, as well as their microbiome structure, may be key factors. SIGNIFICANCE: The astigmatid mites occurring in homes are significant producers of allergens that are highly dangerous to humans and domesticated animals. Mites are tightly associated with microorganisms that affect their biology and consequently allergy signatures. Mite populations were found to be infected with certain intracellular bacteria, but some populations lacked an intracellular bacterium. Our previous research showed that some populations of Tyrophagus putrescentiae are infected with Wolbachia, but some populations host additional bacteria of interest. Thus, there are not only interactions between the mites and Wolbachia but also likely an additional level of interaction that can be found in the interaction between different bacteria in the mites. These "higher-level" signatures and consequences that bacteria affect, including allergen production, are not understood in mites. In this study, we identified Wolbachia-specific proteins in mites for the first time. This study provides Wolbachia- and mite-derived markers that can be clues for describing "higher-level" mite-bacterium-bacterium interactions. Indeed, the microbiome contribution to allergies can potentially be derived directly from bacterial proteins, especially if they are abundant.}, } @article {pmid34436640, year = {2022}, author = {Möhlmann, TWR and Ter Braak, CJF and Te Beest, DE and Hendriks, M and Nijhuis, EH and Warris, S and Drolet, BS and van Overbeek, L and Koenraadt, CJM}, title = {Species Identity, Life History, and Geographic Distance Influence Gut Bacterial Communities in Lab-Reared and European Field-Collected Culicoides Biting midges.}, journal = {Microbial ecology}, volume = {84}, number = {1}, pages = {267-284}, pmid = {34436640}, issn = {1432-184X}, support = {BBS/E/I/00007039/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 1300018161//ANIHWA-EraNet/ ; Global One Health strategic program//Wageningen University/ ; }, mesh = {Animals ; *Ceratopogonidae ; *Gastrointestinal Microbiome ; Insect Vectors/microbiology ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {Bacteria are part of the insect gut system and influence many physiological traits of their host. Gut bacteria may even reduce or block the transmission of arboviruses in several species of arthropod vectors. Culicoides biting midges are important arboviral vectors of several livestock and wildlife diseases, yet limited information is available on their gut bacterial communities. Addressing this gap will help inform how these communities can be manipulated and ultimately used as novel tools to control pathogens. To assess how bacterial communities change during the life stages of lab-reared C. nubeculosus and C. sonorensis, endosymbiotic bacteria were identified using Illumina sequencing of 16S rRNA and taxonomically characterised. Analyses were conducted to determine how gut bacterial communities in adults are influenced by species identity and geographic distance among biting midge populations. Communities of the two lab-reared Culicoides species significantly changed after pupation and with maturation into 6-day-old adults. Pseudomonas, Burkholderiaceae and Leucobacter bacteria were part of a core community that was trans-stadially transmitted and found throughout their life cycle. Among field-collected biting midges, the bacterial communities were unique for almost each species. Cardinium, Rickettsia and Wolbachia were some of the most abundant bacteria in midges collected from wetlands. Only Pseudomonas was present in high relative abundance in all field-collected species. In this study, species identity, as well as geographic distance, influenced the gut bacterial communities and may partly explain known inter- and intra-species variability in vector competence. Additionally, stably associated bacterial species could be candidates for paratransgenic strategies to control vector-borne pathogens.}, } @article {pmid34432527, year = {2021}, author = {Edenborough, KM and Flores, HA and Simmons, CP and Fraser, JE}, title = {Correction for Edenborough et al., "Using Wolbachia to Eliminate Dengue: Will the Virus Fight Back?".}, journal = {Journal of virology}, volume = {95}, number = {18}, pages = {e0095321}, doi = {10.1128/JVI.00953-21}, pmid = {34432527}, issn = {1098-5514}, } @article {pmid34402109, year = {2021}, author = {Arif, S and Gerth, M and Hone-Millard, WG and Nunes, MDS and Dapporto, L and Shreeve, TG}, title = {Evidence for multiple colonisations and Wolbachia infections shaping the genetic structure of the widespread butterfly Polyommatus icarus in the British Isles.}, journal = {Molecular ecology}, volume = {30}, number = {20}, pages = {5196-5213}, doi = {10.1111/mec.16126}, pmid = {34402109}, issn = {1365-294X}, mesh = {Animals ; *Butterflies/genetics ; DNA, Mitochondrial/genetics ; Genetic Structures ; Genetic Variation ; Phylogeny ; Phylogeography ; Refugium ; *Wolbachia/genetics ; }, abstract = {The paradigm of isolation in southern refugia during glacial periods followed by expansions during interglacials, producing limited genetic differentiation in northern areas, dominates European phylogeography. However, the existence of complex structured populations in formerly glaciated areas, and islands connected to mainland areas during glacial maxima, call for alternative explanations. We reconstructed the mtDNA phylogeography of the widespread Polyommatus Icarus butterfly with an emphasis on the formerly glaciated and connected British Isles. We found distinct geographical structuring of CO1 haplogroups, with an ancient lineage restricted to the marginal European areas, including Northern Scotland and Outer Hebrides. Population genomic analyses, using ddRADSeq genomic markers, also reveal substantial genetic structuring within Britain. However, there is negligble mito-nuclear concordance consistent with independent demographic histories of mitochondrial versus nuclear DNA. While mtDNA-Wolbachia associations in northern Britain could account for the geographic structuring of mtDNA across most of the British Isles, for nuclear DNA markers (derived from ddRADseq data) butterflies from France cluster between northern and southern British populations - an observation consistent with a scenario of multiple recolonisation. Taken together our results suggest that contemporary mtDNA structuring in the British Isles (and potentially elsewhere in Europe) largely results from Wolbachia infections, however, nuclear genomic structuring suggests a history of at least two distinct colonisations. This two-stage colonisation scenario has previously been put forth to explain genetic diversity and structuring in other British flora and fauna. Additionally, we also present preliminary evidence for potential Wolbachia-induced feminization in the Outer Hebrides.}, } @article {pmid34394061, year = {2021}, author = {Gesto, JSM and Pinto, SB and Dias, FBS and Peixoto, J and Costa, G and Kutcher, S and Montgomery, J and Green, BR and Anders, KL and Ryan, PA and Simmons, CP and O'Neill, SL and Moreira, LA}, title = {Large-Scale Deployment and Establishment of Wolbachia Into the Aedes aegypti Population in Rio de Janeiro, Brazil.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {711107}, pmid = {34394061}, issn = {1664-302X}, abstract = {Traditional methods of vector control have proven insufficient to reduce the alarming incidence of dengue, Zika, and chikungunya in endemic countries. The bacterium symbiont Wolbachia has emerged as an efficient pathogen-blocking and self-dispersing agent that reduces the vectorial potential of Aedes aegypti populations and potentially impairs arboviral disease transmission. In this work, we report the results of a large-scale Wolbachia intervention in Ilha do Governador, Rio de Janeiro, Brazil. wMel-infected adults were released across residential areas between August 2017 and March 2020. Over 131 weeks, including release and post-release phases, we monitored the wMel prevalence in field specimens and analyzed introgression profiles of two assigned intervention areas, RJ1 and RJ2. Our results revealed that wMel successfully invaded both areas, reaching overall infection rates of 50-70% in RJ1 and 30-60% in RJ2 by the end of the monitoring period. At the neighborhood-level, wMel introgression was heterogeneous in both RJ1 and RJ2, with some profiles sustaining a consistent increase in infection rates and others failing to elicit the same. Correlation analysis revealed a weak overall association between RJ1 and RJ2 (r = 0.2849, p = 0.0236), and an association at a higher degree when comparing different deployment strategies, vehicle or backpack-assisted, within RJ1 (r = 0.4676, p < 0.0001) or RJ2 (r = 0.6263, p < 0.0001). The frequency knockdown resistance (kdr) alleles in wMel-infected specimens from both areas were consistently high over this study. Altogether, these findings corroborate that wMel can be successfully deployed at large-scale as part of vector control intervention strategies and provide the basis for imminent disease impact studies in Southeastern Brazil.}, } @article {pmid34390609, year = {2021}, author = {Towett-Kirui, S and Morrow, JL and Close, S and Royer, JE and Riegler, M}, title = {Host-endoparasitoid-endosymbiont relationships: concealed Strepsiptera provide new twist to Wolbachia in Australian tephritid fruit flies.}, journal = {Environmental microbiology}, volume = {23}, number = {9}, pages = {5587-5604}, doi = {10.1111/1462-2920.15715}, pmid = {34390609}, issn = {1462-2920}, mesh = {Animals ; Australia ; *Symbiosis ; *Tephritidae/microbiology ; *Wolbachia/genetics ; }, abstract = {Wolbachia are widespread endosymbionts that affect arthropod reproduction and fitness. Mostly maternally inherited, Wolbachia are occasionally transferred horizontally. Previously, two Wolbachia strains were reported at low prevalence and titres across seven Australian tephritid species, possibly indicative of frequent horizontal transfer. Here, we performed whole-genome sequencing of field-caught Wolbachia-positive flies. Unexpectedly, we found complete mitogenomes of an endoparasitic strepsipteran, Dipterophagus daci, suggesting that Wolbachia in the flies are linked to concealed parasitization. We performed the first genetic characterization of D. daci and detected D. daci in Wolbachia-positive flies not visibly parasitized, and most but not all Wolbachia-negative flies were D. daci-negative, presumably reflecting polymorphism for the Wolbachia infections in D. daci. We dissected D. daci from stylopized flies and confirmed that Wolbachia infects D. daci, but also found Wolbachia in stylopized fly tissues, likely somatic, horizontally transferred, non-heritable infections. Furthermore, no Wolbachia cif and wmk genes were detected and very low mitogenomic variation in D. daci across its distribution. Therefore, Wolbachia may influence host fitness without reproductive manipulation. Our study of 13 tephritid species highlights that concealed early stages of strepsipteran parasitization led to the previous incorrect assignment of Wolbachia co-infections to tephritid species, obscuring ecological studies of this common endosymbiont and its horizontal transmission by parasitoids.}, } @article {pmid34389828, year = {2022}, author = {Perreau, J and Moran, NA}, title = {Genetic innovations in animal-microbe symbioses.}, journal = {Nature reviews. Genetics}, volume = {23}, number = {1}, pages = {23-39}, pmid = {34389828}, issn = {1471-0064}, support = {R35 GM131738/GM/NIGMS NIH HHS/United States ; }, mesh = {Aliivibrio/*genetics/physiology ; Animals ; Arthropods/*genetics/microbiology ; Decapodiformes/*genetics/microbiology ; Gene Flow ; Genetic Drift ; Host Microbial Interactions/*genetics ; Models, Genetic ; Phylogeny ; Selection, Genetic ; Symbiosis/*genetics ; Wolbachia/classification/*genetics/physiology ; }, abstract = {Animal hosts have initiated myriad symbiotic associations with microorganisms and often have maintained these symbioses for millions of years, spanning drastic changes in ecological conditions and lifestyles. The establishment and persistence of these relationships require genetic innovations on the parts of both symbionts and hosts. The nature of symbiont innovations depends on their genetic population structure, categorized here as open, closed or mixed. These categories reflect modes of inter-host transmission that result in distinct genomic features, or genomic syndromes, in symbionts. Although less studied, hosts also innovate in order to preserve and control symbiotic partnerships. New capabilities to sequence host-associated microbial communities and to experimentally manipulate both hosts and symbionts are providing unprecedented insights into how genetic innovations arise under different symbiont population structures and how these innovations function to support symbiotic relationships.}, } @article {pmid34388986, year = {2021}, author = {Morrow, JL and Riegler, M}, title = {Genome analyses of four Wolbachia strains and associated mitochondria of Rhagoletis cerasi expose cumulative modularity of cytoplasmic incompatibility factors and cytoplasmic hitchhiking across host populations.}, journal = {BMC genomics}, volume = {22}, number = {1}, pages = {616}, pmid = {34388986}, issn = {1471-2164}, mesh = {Animals ; Biological Evolution ; Cytoplasm ; Humans ; Mitochondria ; Symbiosis/genetics ; *Tephritidae ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: The endosymbiont Wolbachia can manipulate arthropod reproduction and invade host populations by inducing cytoplasmic incompatibility (CI). Some host species are coinfected with multiple Wolbachia strains which may have sequentially invaded host populations by expressing different types of modular CI factor (cif) genes. The tephritid fruit fly Rhagoletis cerasi is a model for CI and Wolbachia population dynamics. It is associated with at least four Wolbachia strains in various combinations, with demonstrated (wCer2, wCer4), predicted (wCer1) or unknown (wCer5) CI phenotypes.

RESULTS: We sequenced and assembled the draft genomes of the Wolbachia strains wCer1, wCer4 and wCer5, and compared these with the previously sequenced genome of wCer2 which currently invades R. cerasi populations. We found complete cif gene pairs in all strains: four pairs in wCer2 (three Type I; one Type V), two pairs in wCer1 (both Type I) and wCer4 (one Type I; one Type V), and one pair in wCer5 (Type IV). Wolbachia genome variant analyses across geographically and genetically distant host populations revealed the largest diversity of single nucleotide polymorphisms (SNPs) in wCer5, followed by wCer1 and then wCer2, indicative of their different lengths of host associations. Furthermore, mitogenome analyses of the Wolbachia genome-sequenced individuals in combination with SNP data from six European countries revealed polymorphic mitogenome sites that displayed reduced diversity in individuals infected with wCer2 compared to those without.

CONCLUSIONS: Coinfections with Wolbachia are common in arthropods and affect options for Wolbachia-based management strategies of pest and vector species already infected by Wolbachia. Our analyses of Wolbachia genomes of a host naturally coinfected by several strains unravelled signatures of the evolutionary dynamics in both Wolbachia and host mitochondrial genomes as a consequence of repeated invasions. Invasion of already infected populations by new Wolbachia strains requires new sets of functionally different cif genes and thereby may select for a cumulative modularity of cif gene diversity in invading strains. Furthermore, we demonstrated at the mitogenomic scale that repeated CI-driven Wolbachia invasions of hosts result in reduced mitochondrial diversity and hitchhiking effects. Already resident Wolbachia strains may experience similar cytoplasmic hitchhiking effects caused by the invading Wolbachia strain.}, } @article {pmid34383852, year = {2021}, author = {Zhang, HB and Cao, Z and Qiao, JX and Zhong, ZQ and Pan, CC and Liu, C and Zhang, LM and Wang, YF}, title = {Metabolomics provide new insights into mechanisms of Wolbachia-induced paternal defects in Drosophila melanogaster.}, journal = {PLoS pathogens}, volume = {17}, number = {8}, pages = {e1009859}, pmid = {34383852}, issn = {1553-7374}, mesh = {Animals ; Bacterial Infections/*complications/metabolism/microbiology ; Drosophila melanogaster/growth & development/*metabolism/microbiology ; Female ; Infertility, Male/etiology/metabolism/*pathology ; Male ; *Metabolome ; *Phenotype ; *Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a group of intracellular symbiotic bacteria that widely infect arthropods and nematodes. Wolbachia infection can regulate host reproduction with the most common phenotype in insects being cytoplasmic incompatibility (CI), which results in embryonic lethality when uninfected eggs fertilized with sperms from infected males. This suggests that CI-induced defects are mainly in paternal side. However, whether Wolbachia-induced metabolic changes play a role in the mechanism of paternal-linked defects in embryonic development is not known. In the current study, we first use untargeted metabolomics method with LC-MS to explore how Wolbachia infection influences the metabolite profiling of the insect hosts. The untargeted metabolomics revealed 414 potential differential metabolites between Wolbachia-infected and uninfected 1-day-old (1d) male flies. Most of the differential metabolites were significantly up-regulated due to Wolbachia infection. Thirty-four metabolic pathways such as carbohydrate, lipid and amino acid, and vitamin and cofactor metabolism were affected by Wolbachia infection. Then, we applied targeted metabolomics analysis with GC-MS and showed that Wolbachia infection resulted in an increased energy expenditure of the host by regulating glycometabolism and fatty acid catabolism, which was compensated by increased food uptake. Furthermore, overexpressing two acyl-CoA catabolism related genes, Dbi (coding for diazepam-binding inhibitor) or Mcad (coding for medium-chain acyl-CoA dehydrogenase), ubiquitously or specially in testes caused significantly decreased paternal-effect egg hatch rate. Oxidative stress and abnormal mitochondria induced by Wolbachia infection disrupted the formation of sperm nebenkern. These findings provide new insights into mechanisms of Wolbachia-induced paternal defects from metabolic phenotypes.}, } @article {pmid34379678, year = {2021}, author = {Fakhour, S and Renoz, F and Ambroise, J and Pons, I and Noël, C and Gala, JL and Hance, T}, title = {Insight into the bacterial communities of the subterranean aphid Anoecia corni.}, journal = {PloS one}, volume = {16}, number = {8}, pages = {e0256019}, pmid = {34379678}, issn = {1932-6203}, mesh = {Animals ; Aphids/*microbiology ; Bacteria/*classification/genetics/*isolation & purification ; *Biodiversity ; *Biological Evolution ; *Genetic Variation ; High-Throughput Nucleotide Sequencing ; Microbiota ; Morocco ; Phylogeny ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Many insect species are associated with bacterial partners that can significantly influence their evolutionary ecology. Compared to other insect groups, aphids harbor a bacterial microbiota that has the reputation of being poorly diversified, generally limited to the presence of the obligate nutritional symbiont Buchnera aphidicola and some facultative symbionts. In this study, we analyzed the bacterial diversity associated with the dogwood-grass aphid Anoecia corni, an aphid species that spends much of its life cycle in a subterranean environment. Little is known about the bacterial diversity associated with aphids displaying such a lifestyle, and one hypothesis is that close contact with the vast microbial community of the rhizosphere could promote the acquisition of a richer bacterial diversity compared to other aphid species. Using 16S rRNA amplicon Illumina sequencing on specimens collected on wheat roots in Morocco, we identified 10 bacterial operational taxonomic units (OTUs) corresponding to five bacterial genera. In addition to the obligate symbiont Buchnera, we identified the facultative symbionts Serratia symbiotica and Wolbachia in certain aphid colonies. The detection of Wolbachia is unexpected as it is considered rare in aphids. Moreover, its biological significance remains unknown in these insects. Besides, we also detected Arsenophonus and Dactylopiibacterium carminicum. These results suggest that, despite its subterranean lifestyle, A. corni shelter a bacterial diversity mainly limited to bacterial endosymbionts.}, } @article {pmid34379518, year = {2021}, author = {Ross, PA and Gu, X and Robinson, KL and Yang, Q and Cottingham, E and Zhang, Y and Yeap, HL and Xu, X and Endersby-Harshman, NM and Hoffmann, AA}, title = {A wAlbB Wolbachia Transinfection Displays Stable Phenotypic Effects across Divergent Aedes aegypti Mosquito Backgrounds.}, journal = {Applied and environmental microbiology}, volume = {87}, number = {20}, pages = {e0126421}, pmid = {34379518}, issn = {1098-5336}, mesh = {Aedes/*microbiology ; Animals ; Female ; Heat-Shock Response ; Male ; Phenotype ; Transfection ; Wolbachia/*genetics ; }, abstract = {Aedes mosquitoes harboring intracellular Wolbachia bacteria are being released in arbovirus and mosquito control programs. With releases taking place around the world, understanding the contribution of host variation to Wolbachia phenotype is crucial. We generated a Wolbachia transinfection (wAlbB[Q]) in Aedes aegypti and performed backcrossing to introduce the infection into Australian or Malaysian nuclear backgrounds. Whole Wolbachia genome sequencing shows that the wAlbB[Q] transinfection is nearly identical to the reference wAlbB genome, suggesting few changes since the infection was first introduced to A. aegypti over 15 years ago. However, these sequences were distinct from other available wAlbB genome sequences, highlighting the potential diversity of wAlbB in natural Aedes albopictus populations. Phenotypic comparisons demonstrate the effects of wAlbB infection on egg hatching and nuclear background on fecundity and body size but no interactions between wAlbB infection and nuclear background for any trait. The wAlbB infection was stable at high temperatures and showed perfect maternal transmission and cytoplasmic incompatibility regardless of the host background. Our results demonstrate the stability of wAlbB across host backgrounds and point to its long-term effectiveness for controlling arbovirus transmission and mosquito populations. IMPORTANCEWolbachia bacteria are being used to control the transmission of dengue virus and other arboviruses by mosquitoes. For Wolbachia release programs to be effective globally, Wolbachia infections must be stable across mosquito populations from different locations. In this study, we transferred Wolbachia (strain wAlbB) to Aedes aegypti mosquitoes with an Australian genotype and introduced the infection to Malaysian mosquitoes through backcrossing. We found that the phenotypic effects of Wolbachia are stable across both mosquito backgrounds. We sequenced the genome of wAlbB and found very few genetic changes despite spending over 15 years in a novel mosquito host. Our results suggest that the effects of Wolbachia infections are likely to remain stable across time and host genotype.}, } @article {pmid34367558, year = {2021}, author = {Wheeler, TB and Thompson, V and Conner, WR and Cooper, BS}, title = {Wolbachia in the spittlebug Prosapia ignipectus: Variable infection frequencies, but no apparent effect on host reproductive isolation.}, journal = {Ecology and evolution}, volume = {11}, number = {15}, pages = {10054-10065}, pmid = {34367558}, issn = {2045-7758}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, abstract = {Animals serve as hosts for complex communities of microorganisms, including endosymbionts that live inside their cells. Wolbachia bacteria are perhaps the most common endosymbionts, manipulating host reproduction to propagate. Many Wolbachia cause cytoplasmic incompatibility (CI), which results in reduced egg hatch when uninfected females mate with infected males. Wolbachia that cause intense CI spread to high and relatively stable frequencies, while strains that cause weak or no CI tend to persist at intermediate, often variable, frequencies. Wolbachia could also contribute to host reproductive isolation (RI), although current support for such contributions is limited to a few systems. To test for Wolbachia frequency variation and effects on host RI, we sampled several local Prosapia ignipectus (Fitch) (Hemiptera: Cercopidae) spittlebug populations in the northeastern United States over two years, including closely juxtaposed Maine populations with different monomorphic color forms, "black" and "lined." We discovered a group-B Wolbachia (wPig) infecting P. ignipectus that diverged from group-A Wolbachia-like model wMel and wRi strains in Drosophila-6 to 46 MYA. Populations of the sister species Prosapia bicincta (Say) from Hawaii and Florida are uninfected, suggesting that P. ignipectus acquired wPig after their initial divergence. wPig frequencies were generally high and variable among sites and between years. While phenotyping wPig effects on host reproduction is not currently feasible, the wPig genome contains three divergent sets of CI loci, consistent with high wPig frequencies. Finally, Maine monomorphic black and monomorphic lined populations of P. ignipectus share both wPig and mtDNA haplotypes, implying no apparent effect of wPig on the maintenance of this morphological contact zone. We hypothesize P. ignipectus acquired wPig horizontally as observed for many Drosophila species, and that significant CI and variable transmission produce high but variable wPig frequencies.}, } @article {pmid34365209, year = {2021}, author = {Li, XD and Xin, L and Rong, WT and Liu, XY and Deng, WA and Qin, YC and Li, XL}, title = {Effect of heavy metals pollution on the composition and diversity of the intestinal microbial community of a pygmy grasshopper (Eucriotettix oculatus).}, journal = {Ecotoxicology and environmental safety}, volume = {223}, number = {}, pages = {112582}, doi = {10.1016/j.ecoenv.2021.112582}, pmid = {34365209}, issn = {1090-2414}, mesh = {Animals ; *Grasshoppers ; *Metals, Heavy/analysis/toxicity ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Soil ; Soil Microbiology ; *Soil Pollutants/analysis/toxicity ; }, abstract = {Heavy metal pollution in the environment is mostly manifested as a multi-elemental compound pollution. The effect of the long-term exposure to heavy metal pollution on the gut microbes of insects has remained unknown. For the current work, the population of Eucriotettix oculatus living in mining areas around the Diaojiang River with a history of hundreds of years of pollution, was selected along with the similar species living in non-mining areas to conduct a comparative study of their gut microbes. The microbial communities were analyzed using 16S rRNA amplicon sequencing. The results showed Proteobacteria to be dominant among gut microbes of E. oculatus, but the abundance of Proteobacteria was significantly increased when the insects were exposed to the environment with heavy metal pollution. The symbiotic bacteria belonging to genus Wolbachia were found to be dominant among the insect population from the non-mining area group, while the pathogenic bacteria belonging to Aeromonas were dominant among the insect population of the mining area group. The diversity analysis showed that the gut microbial community diversity of E. oculatus was reduced in the heavy metal pollution habitat. The analysis of the differences in the gut microbial population and metabolic pathways of the two groups showed that the heavy metal pollution caused the increase in pathogenic bacteria among the gut microbes of E. oculatus, which might have a negative impact on the health of the host. At the same time, probiotics and the beneficial metabolism pathways were also found to increase and enhance, helping the host to resist the damage caused by heavy metal stress. This might be one of the strategies used by E. oculatus to adapt to heavy metal pollution.}, } @article {pmid34364896, year = {2021}, author = {Mendoza-Roldan, JA and Gabrielli, S and Cascio, A and Manoj, RRS and Bezerra-Santos, MA and Benelli, G and Brianti, E and Latrofa, MS and Otranto, D}, title = {Zoonotic Dirofilaria immitis and Dirofilaria repens infection in humans and an integrative approach to the diagnosis.}, journal = {Acta tropica}, volume = {223}, number = {}, pages = {106083}, doi = {10.1016/j.actatropica.2021.106083}, pmid = {34364896}, issn = {1873-6254}, mesh = {Animals ; *Dirofilaria immitis/genetics ; *Dirofilaria repens/genetics ; *Dirofilariasis/diagnosis/epidemiology ; Dog Diseases/diagnosis/epidemiology ; Dogs ; Humans ; Islands ; Italy ; Phylogeny ; Seroepidemiologic Studies ; *Zoonoses/diagnosis/parasitology ; }, abstract = {Dirofilariosis by Dirofilaria immitis and Dirofilaria repens is endemic in dogs from countries of the Mediterranean basin. Both species may infect humans, with most of the infected patients remaining asymptomatic. Based on the recent description of the southernmost hyperendemic European focus of heartworm disease in dogs from the Pelagie archipelagos, we performed a serological and molecular survey in human population of that area. Human blood samples were collected in the islands of Linosa (n=101) and Lampedusa (n=296) and tested by ELISA and molecular test for the detection of D. immitis and D. repens. Samples were also screened for filarioid-associated endosymbionts, Wolbachia sp. The seroprevalence of D. immitis and D. repens was, respectively, 7.9% and 3.96% in Linosa, and 7.77% and 19.93% in Lampedusa. Out of 397 human blood samples tested molecularly, 4 scored positive (1%) for Dirofilaria spp. by qPCR (i.e., three for D. immitis and one for D. repens) and 6 (1.5%) for Wolbachia. Of the qPCR positive for Dirofilaria spp., only D. repens was amplified by cPCR and was positive for Wolbachia. In the phylogenetic analysis, the sequence of Wolbachia detected in D. repens positive samples clustered along with other C supergroup filarioids. Our results overlap with the recent prevalence data collected on dogs from the same area, where D. immitis is prevalent in Linosa and D. repens prevails in Lampedusa. Molecular detection of D. immitis in human blood is quite unusual considering that humans are dead-end hosts for dirofilarial infection and most of the human cases described so far in Europe were ascribed to D. repens. An integrative diagnostic approach using serum analysis and Wolbachia detection is also presented. In endemic areas for canine dirofilarioses humans are exposed to the infection, suggesting the importance of One Health approach in diagnosing, treating and controlling this zoonotic parasitosis.}, } @article {pmid34351459, year = {2021}, author = {Balaji, S and Deepthi, KNG and Prabagaran, SR}, title = {Native Wolbachia influence bacterial composition in the major vector mosquito Aedes aegypti.}, journal = {Archives of microbiology}, volume = {203}, number = {8}, pages = {5225-5240}, pmid = {34351459}, issn = {1432-072X}, mesh = {*Aedes ; Animals ; Female ; Mosquito Vectors ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {Bacterial species that inhabit mosquito microbiota play an essential role in determining vector competence. In addition to critical factors such as host genotype, feeding habit and geography, intracellular endosymbiont Wolbachia pipientis modulates microbial composition considerably. In the present study, we assessed the midgut bacterial diversity of Aedes aegypti mosquitoes that is either naturally carrying Wolbachia (wAegB[+]) or antibiotic cured (wAegB[-]) through a culture-independent approach. Towards this, 16S rRNA gene libraries were constructed from midgut bacterial DNA of laboratory-reared larvae and adult female mosquitoes fed with sugar or blood. Among them 33 genera comprising 65 distinct species were identified, where > 75% of bacterial taxa were commonly shared by both groups (wAegB[+] and wAegB[-]), implying a subtle shift in the bacterial composition influenced by Wolbachia. Though the change was mostly restricted to minimally represented species, predominant taxa were observed unaltered except for certain genera. While Serratia sp. was abundant in Wolbachia carrying mosquitoes, Pseudomonas sp. and Acinetobacter sp. were predominant in Wolbachia free mosquitoes. This result demonstrates the influence of Wolbachia that could modulate the colonization of certain resident bacterial taxa through competitive interactions. Overall, this study shed more light on the impact of wAegB in altering the gut microbiota of Ae. aegypti mosquito, which might challenge host fitness and vector competence.}, } @article {pmid34350357, year = {2021}, author = {Ma, Y and Huang, CW and Luan, YX and Chen, WJ}, title = {The mitochondrial genome of a minute springtail species Megalothorax incertus (Collembola: Neelipleona: Neelidae).}, journal = {Mitochondrial DNA. Part B, Resources}, volume = {6}, number = {8}, pages = {2430-2432}, pmid = {34350357}, issn = {2380-2359}, abstract = {In this study, the complete mitochondrial genome for the Wolbachia infected parthenogenetic collembola Megalothorax incertus Börner, 1903 was determined. It represents the first report of a complete mitochondrial genome from Neelipleona, one of the four orders of Collembola. The circularized 14,994 bp mitochondrial genome sequence consists of canonical 37 mito-genes, including 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes. The base composition of the majority strand (same direction as most of the mitochondrial genes) is 32.0% for A, 24.1% for C, 11.9% for G, and 32.1% for T. The phylogenetic trees inferred from 13 PCGs using the Maximum-likelihood (ML) and Bayesian inference (BI) methods suggested that Neelidae is basal to the remaining springtails. This mitochondrial genome provides new insights to decipher the phylogeny of Collembola.}, } @article {pmid34349742, year = {2021}, author = {Zhang, Y and Liu, S and Jiang, R and Zhang, C and Gao, T and Wang, Y and Liu, C and Long, Y and Zhang, Y and Yang, Y}, title = {Wolbachia Strain wGri From the Tea Geometrid Moth Ectropis grisescens Contributes to Its Host's Fecundity.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {694466}, pmid = {34349742}, issn = {1664-302X}, abstract = {Members of the Wolbachia genus manipulate insect-host reproduction and are the most abundant bacterial endosymbionts of insects. The tea Geometrid moth Ectropis grisescens (Warren) (Lepidoptera: Geometridae) is the most devastating insect pest of tea plants [Camellia sinensis (L.) O. Kuntze] in China. However, limited data on the diversity, typing, or phenotypes of Wolbachia in E. grisescens are available. Here, we used a culture-independent method to compare the gut bacteria of E. grisescens and other tea Geometridae moths. The results showed that the composition of core gut bacteria in larvae of the three Geometridae moth species was similar, except for the presence of Wolbachia. Moreover, Wolbachia was also present in adult female E. grisescens samples. A Wolbachia strain was isolated from E. grisescens and designated as wGri. Comparative analyses showed that this strain shared multilocus sequence types and Wolbachia surface protein hypervariable region profiles with cytoplasmic incompatibility (CI)-inducing strains in supergroup B; however, the wGri-associated phenotypes were undetermined. A reciprocal cross analysis showed that Wolbachia-uninfected females mated with infected males resulted in 100% embryo mortality (0% eggs hatched per female). Eggs produced by mating between uninfected males and infected females hatched normally. These findings indicated that wGri induces strong unidirectional CI in E. grisescens. Additionally, compared with uninfected females, Wolbachia-infected females produced approximately 30-40% more eggs. Together, these results show that this Wolbachia strain induces reproductive CI in E. grisescens and enhances the fecundity of its female host. We also demonstrated that wGri potential influences reproductive communication between E. grisescens and Ectropis obliqua through CI.}, } @article {pmid34346878, year = {2021}, author = {Karatepe, M and Aksoy, S and Karatepe, B}, title = {Wolbachia spp. and Spiroplasma spp. in Musca spp.: Detection Using Molecular Approaches.}, journal = {Turkiye parazitolojii dergisi}, volume = {45}, number = {3}, pages = {211-215}, doi = {10.4274/tpd.galenos.2021.35229}, pmid = {34346878}, issn = {2146-3077}, mesh = {Animals ; Female ; Male ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; *Spiroplasma/genetics ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {OBJECTIVE: This study aimed to detect the presence of Wolbachia and Spiroplasma endosymbionts in Musca flies through molecular approaches.

METHODS: In total, 40 Musca spp. (20 female and 20 male) were used. Before DNA extraction, the flies were dissected and their heads, wings and legs were detached from their bodies under a stereomicroscope. Genomic DNA was analysed by standard polymerase chain reaction (PCR) using primers against Musca beta-tubulin. Afterward, the samples were examined for the presence of Wolbachia spp. using primers against Wolbachia wsp and GroEL. Furthermore, the DNA samples were analysed by PCR to detect the presence of Spiroplasma using primers against the 16S rRNA.

RESULTS: No Wolbachia positivity was detected in Musca flies, as shown by the negative PCR results for wsp and GroEL. Spiroplasma positivity was detected in 5% (1/20) of the female Musca flies but not in the male flies (0/20).

CONCLUSION: Wolbachia spp. were not detected in Musca flies. Of the total Musca flies, only one was positive for Spiroplasma spp. To our knowledge, this is the first study to detect the presence of Spiroplasma in Musca flies.}, } @article {pmid34324610, year = {2021}, author = {Rothman, JA and Loope, KJ and McFrederick, QS and Wilson Rankin, EE}, title = {Microbiome of the wasp Vespula pensylvanica in native and invasive populations, and associations with Moku virus.}, journal = {PloS one}, volume = {16}, number = {7}, pages = {e0255463}, pmid = {34324610}, issn = {1932-6203}, mesh = {Animals ; *Bees ; RNA Viruses ; *RNA, Ribosomal, 16S ; *Wasps ; }, abstract = {Invasive species present a worldwide concern as competition and pathogen reservoirs for native species. Specifically, the invasive social wasp, Vespula pensylvanica, is native to western North America and has become naturalized in Hawaii, where it exerts pressures on native arthropod communities as a competitor and predator. As invasive species may alter the microbial and disease ecology of their introduced ranges, there is a need to understand the microbiomes and virology of social wasps. We used 16S rRNA gene sequencing to characterize the microbiome of V. pensylvanica samples pooled by colony across two geographically distinct ranges and found that wasps generally associate with taxa within the bacterial genera Fructobacillus, Fructilactobacillus, Lactococcus, Leuconostoc, and Zymobacter, and likely associate with environmentally-acquired bacteria. Furthermore, V. pensylvanica harbors-and in some cases were dominated by-many endosymbionts including Wolbachia, Sodalis, Arsenophonus, and Rickettsia, and were found to contain bee-associated taxa, likely due to scavenging on or predation upon honey bees. Next, we used reverse-transcriptase quantitative PCR to assay colony-level infection intensity for Moku virus (family: Iflaviridae), a recently-described disease that is known to infect multiple Hymenopteran species. While Moku virus was prevalent and in high titer, it did not associate with microbial diversity, indicating that the microbiome may not directly interact with Moku virus in V. pensylvanica in meaningful ways. Collectively, our results suggest that the invasive social wasp V. pensylvanica associates with a simple microbiome, may be infected with putative endosymbionts, likely acquires bacterial taxa from the environment and diet, and is often infected with Moku virus. Our results suggest that V. pensylvanica, like other invasive social insects, has the potential to act as a reservoir for bacteria pathogenic to other pollinators, though this requires experimental demonstration.}, } @article {pmid34314434, year = {2021}, author = {Dutra, HLC and Ford, SA and Allen, SL and Bordenstein, SR and Chenoweth, SF and Bordenstein, SR and McGraw, EA}, title = {The impact of artificial selection for Wolbachia-mediated dengue virus blocking on phage WO.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {7}, pages = {e0009637}, pmid = {34314434}, issn = {1935-2735}, support = {R01 AI143725/AI/NIAID NIH HHS/United States ; R01 AI143758/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; *Bacteriophages ; Biological Control Agents ; Dengue Virus/*physiology ; *Host-Pathogen Interactions ; Mosquito Vectors ; Mutation ; *Prophages ; Viral Load ; Wolbachia/*physiology ; }, abstract = {Wolbachia is currently at the forefront of global efforts to control arbovirus transmission from the vector Aedes aegypti. The use of Wolbachia relies on two phenotypes-cytoplasmic incompatibility (CI), conferred by cifA and cifB genes in prophage WO, and Wolbachia-mediated pathogen blocking (WMPB). These traits allow for local, self-sustaining reductions in transmission of dengue (DENV) following release of Wolbachia-infected A. aegypti. Here, aided by previous artificial selection experiment that generated Low and High pathogen blocking lines, we examined the potential link between WMPB and phage WO. We found no evidence that Wolbachia or phage WO relative densities predict DENV blocking strength across selected lines. However, selection resulted in reduced phage WO relative density for the Low WMPB line. The Low blocking line was previously shown to have reduced fitness as a result of selection. Through subsequent genomic analyses, we demonstrate that SNP variation underpinning selection for low blocking led to elevated frequency of potential deleterious SNPs on chromosome 1. The key region on chromosome 1 contains genes relating to cell cycle regulation, oxidative stress, transcriptional pausing, among others, that may have cascading effects on Wolbachia intracellular environment. We hypothesize that reduction in phage WO may be driven by changes in the loci directly under selection for blocking, or by the accumulation of predicted deleterious alleles in linkage disequilibrium with blocking loci resulting from hitchhiking. For the Low line with fewer phage WO, we also detected reduced expression of cifA and cifB CI genes, with patterns of expression varying between somatic and reproductive tissues. In conclusion, we propose that artificial selection for WMPB trait had corresponding impacts on phage WO densities, and also the transcription of CI-causing genes. Future studies may include a more detailed analysis of the regions the A. aegypti chromosome 1's ability to affect WMPB and other Wolbachia-associated intrinsic factors such as phage WO.}, } @article {pmid34313556, year = {2021}, author = {Schairer, CE and Najera, J and James, AA and Akbari, OS and Bloss, CS}, title = {Oxitec and MosquitoMate in the United States: lessons for the future of gene drive mosquito control.}, journal = {Pathogens and global health}, volume = {115}, number = {6}, pages = {365-376}, pmid = {34313556}, issn = {2047-7732}, mesh = {Animals ; *Gene Drive Technology ; Mosquito Control ; United States ; *Wolbachia ; }, abstract = {In response to growing concerns regarding mosquito-borne diseases, scientists are developing novel systems of vector control. Early examples include Oxitec's OX513A genetically-engineered mosquito and MosquitoMate's Wolbachia-infected mosquito, and systems using 'gene-drive' are in development. Systems based on genetic engineering are controversial and institutions around the world are grappling with the question of who should have a say in how such technologies are field-tested and used. Based on media coverage and public records, we created comparative timelines of the efforts of Oxitec and MosquitoMate to navigate federal and local governance and bring their products to market in the United States. We analyze these timelines with particular attention to the role of public input in technology governance. These cases illustrate how governance of technology in the US is diverse, complex, and opaque. Further, the public response to proposed field trials of the Oxitec product highlights inconsistencies between public expectations for governance and actual practice. As gene-drive mosquito control products develop, both federal and local agencies will find their legitimacy tested without a better procedure for transparently integrating public input.}, } @article {pmid34312980, year = {2021}, author = {Herran, B and Houdelet, C and Raimond, M and Delaunay, C and Cerveau, N and Debenest, C and Grève, P and Bertaux, J}, title = {Feminising Wolbachia disrupt Armadillidium vulgare insulin-like signalling pathway.}, journal = {Cellular microbiology}, volume = {23}, number = {11}, pages = {e13381}, doi = {10.1111/cmi.13381}, pmid = {34312980}, issn = {1462-5822}, support = {//European Regional Development Fund/ ; //National Centre for Scientific Research/ ; }, mesh = {Animals ; Female ; Feminization ; Humans ; Insulin ; *Isopoda ; Male ; Signal Transduction ; *Wolbachia/genetics ; }, abstract = {The endosymbiont Wolbachia feminises male isopods by making them refractory to the insulin-like masculinising hormone, which shunts the autocrine development of the androgenic glands. It was, therefore, proposed that Wolbachia silences the IR receptors, either by preventing their expression or by inactivating them. We describe here the two IR paralogs of Armadillidium vulgare. They displayed a conventional structure and belonged to a family widespread among isopods. Av-IR1 displayed an ubiquist expression, whereas the expression of Av-IR2 was restricted to the gonads. Both were constitutively expressed in males and females and throughout development. However, upon silencing, altered gland physiology and gene expression therein suggested antagonistic roles for Av-IR1 (androinhibiting) and Av-IR2 (androstimulating). They may function in tandem with regulating neurohormones, as a conditional platform that conveys insulin signalling. Wolbachia infection did not alter their expression patterns: leaving the IRs unscathed, the bacteria would suppress the secretion of the neurohormones, thus inducing body-wide IR deactivation and feminisation. Adult males injected with Wolbachia acquired an intersexed physiology. Their phenotypes and gene expressions mirrored the silencing of Av-IR1 only, suggesting that imperfect feminisation stems from a flawed invasion of the androstimulating centre, whereas in fully feminised males invasion would be complete in early juveniles. TAKE AWAY: Two antagonistic Insulin Receptors were characterised in Armadillidium vulgare. The IRs were involved in androstimulating and androinhibiting functions. Wolbachia-induced feminisation did not prevent the expression of the IRs. Imperfectly feminised intersexes phenocopied the silencing of Av-IR1 only. Wolbachia would deactivate the IRs by suppressing neurosecretory co-factors.}, } @article {pmid34305877, year = {2021}, author = {Zhu, YX and Song, ZR and Zhang, YY and Hoffmann, AA and Hong, XY}, title = {Spider Mites Singly Infected With Either Wolbachia or Spiroplasma Have Reduced Thermal Tolerance.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {706321}, pmid = {34305877}, issn = {1664-302X}, abstract = {Heritable symbionts play an essential role in many aspects of host ecology in a temperature-dependent manner. However, how temperature impacts the host and their interaction with endosymbionts remains largely unknown. Here, we investigated the impact of moderate (20°C) and high (30 and 35°C) temperatures on symbioses between the spider mite Tetranychus truncatus and two maternally inherited endosymbionts (Wolbachia and Spiroplasma). We found that the thermal tolerance of mites (as measured by survival after heat exposure) was lower for mites that were singly infected with either Wolbachia or Spiroplasma than it was for co-infected or uninfected mites. Although a relatively high temperature (30°C) is thought to promote bacterial replication, rearing at high temperature (35°C) resulted in losses of Wolbachia and particularly Spiroplasma. Exposing the mites to 20°C reduced the density and transmission of Spiroplasma but not Wolbachia. The four spider mite strains tested differed in the numbers of heat shock genes (Hsps) induced under moderate or high temperature exposure. In thermal preference (Tp) assays, the two Wolbachia-infected spider mite strains preferred a lower temperature than strains without Wolbachia. Our results show that endosymbiont-mediated spider mite responses to temperature stress are complex, involving a combination of changing endosymbiont infection patterns, altered thermoregulatory behavior, and transcription responses.}, } @article {pmid34303627, year = {2021}, author = {Caragata, EP and Dutra, HLC and Sucupira, PHF and Ferreira, AGA and Moreira, LA}, title = {Wolbachia as translational science: controlling mosquito-borne pathogens.}, journal = {Trends in parasitology}, volume = {37}, number = {12}, pages = {1050-1067}, doi = {10.1016/j.pt.2021.06.007}, pmid = {34303627}, issn = {1471-5007}, mesh = {*Aedes ; Animals ; Mosquito Vectors ; Pest Control, Biological ; Translational Science, Biomedical ; *Wolbachia ; }, abstract = {In this review we examine how exploiting the Wolbachia-mosquito relationship has become an increasingly popular strategy for controlling arbovirus transmission. Field deployments of Wolbachia-infected mosquitoes have led to significant decreases in dengue virus incidence via high levels of mosquito population suppression and replacement, emphasizing the success of Wolbachia approaches. Here, we examine how improved knowledge of Wolbachia-host interactions has provided key insight into the mechanisms of the essential phenotypes of pathogen blocking and cytoplasmic incompatibility. And we discuss recent studies demonstrating that extrinsic factors, such as ambient temperature, can modulate Wolbachia density and maternal transmission. Finally, we assess the prospects of using Wolbachia to control other vectors and agricultural pest species.}, } @article {pmid34295000, year = {2021}, author = {Cano-Calle, D and Saldamando-Benjumea, CI and Vivero-Gómez, RJ and Moreno-Herrera, CX and Arango-Isaza, RE}, title = {Two New Strains of Wolbachia Affecting Natural Avocado Thrips.}, journal = {Indian journal of microbiology}, volume = {61}, number = {3}, pages = {348-354}, pmid = {34295000}, issn = {0046-8991}, abstract = {UNLABELLED: Wolbachia is an obligate intracellular bacterium with a high frequency of infection and a continental distribution in arthropods and nematodes. This endosymbiont can induce various reproductive phenotypes in their hosts and has been previously found naturally in several pests including thrips (Thripidae). These insects cause physical fruit damage and economic losses in avocado. The presence of Wolbachia was evaluated for the first time in avocado thrips populations of Frankliniella sp. and Scirtothrips hansoni sp.n. from eastern Antioquia. DNA from adult thrips individuals was used to assess the detection of Wolbachia by amplifying a fragment (600 bp) of the Wolbachia major surface protein (wsp) gene. Results confirmed the presence of two new Wolbachia strains in these two thrips species, with a higher percentage of natural infection in S. hansoni sp.n. The first Wolbachia species was found in Frankliniella sp. and belongs to supergroup A and the second was detected in S. hansoni sp.n. and is part of supergroup B. Wolbachia was more frequently found in females (32.73%), and only found in one male. Analysis of phylogenetic relationships, suggests that the two new Wolbachia sequences (wFran: Frankliniella and wShan: Scirtothrips hansoni) detected here represent two new groups for this endosymbiont. The haplotype network shows the presence of two possible haplotypes for each strain. Future studies to evaluate the possible use of Wolbachia as a control agent in avocado thrips are necessary.

SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00951-5.}, } @article {pmid34293581, year = {2021}, author = {Krueger, S and Moritz, G}, title = {Sperm ultrastructure in arrhenotokous and thelytokous Thysanoptera.}, journal = {Arthropod structure & development}, volume = {64}, number = {}, pages = {101084}, doi = {10.1016/j.asd.2021.101084}, pmid = {34293581}, issn = {1873-5495}, mesh = {Animals ; Female ; Insecta ; Male ; Parthenogenesis ; Reproduction ; Spermatozoa ; *Thysanoptera ; }, abstract = {Thysanoptera are haplo-diploid insects that reproduce either via arrhenotoky or thelytoky. Beside genetically based thelytoky, this reproduction mode can also be endosymbiont induced. The recovery of these females from their infection again leads to the development of males. Functionality of these males ranges widely, and this might be associated with sperm structure. We analyzed the sperm ultrastructure in three different species belonging to both suborders with different reproduction systems via electron microscopy. Beside the different reproduction modes, and adaptations to their life style, the arrhenotokous species Suocerathrips linguis (Thysanoptera: Tubulifera) and Echinothrips americanus (Thysanoptera: Terebrantia) possess typical thysanopteran-like sperm structure. But endosymbiont-cured males from the thelytokous species Hercinothrips femoralis (Thysanoptera: Terebrantia) possess several malformed spermatozoa and a large amount of secretions in their testes. Spermiophagy seems to be typical. It indicates a highly conserved mechanism of the male developmental pathways, despite the observed decay. However, this decay would explain why in some species no stable arrhenotokous line can be re-established.}, } @article {pmid34292940, year = {2021}, author = {Ware-Gilmore, F and Sgrò, CM and Xi, Z and Dutra, HLC and Jones, MJ and Shea, K and Hall, MD and Thomas, MB and McGraw, EA}, title = {Microbes increase thermal sensitivity in the mosquito Aedes aegypti, with the potential to change disease distributions.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {7}, pages = {e0009548}, pmid = {34292940}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*physiology/virology ; Animals ; Dengue Virus/physiology ; Ecosystem ; Hot Temperature ; Mosquito Vectors/*microbiology/*physiology/virology ; Thermotolerance ; Wolbachia/physiology ; }, abstract = {The mosquito Aedes aegypti is the primary vector of many disease-causing viruses, including dengue (DENV), Zika, chikungunya, and yellow fever. As consequences of climate change, we expect an increase in both global mean temperatures and extreme climatic events. When temperatures fluctuate, mosquito vectors will be increasingly exposed to temperatures beyond their upper thermal limits. Here, we examine how DENV infection alters Ae. aegypti thermotolerance by using a high-throughput physiological 'knockdown' assay modeled on studies in Drosophila. Such laboratory measures of thermal tolerance have previously been shown to accurately predict an insect's distribution in the field. We show that DENV infection increases thermal sensitivity, an effect that may ultimately limit the geographic range of the virus. We also show that the endosymbiotic bacterium Wolbachia pipientis, which is currently being released globally as a biological control agent, has a similar impact on thermal sensitivity in Ae. aegypti. Surprisingly, in the coinfected state, Wolbachia did not provide protection against DENV-associated effects on thermal tolerance, nor were the effects of the two infections additive. The latter suggests that the microbes may act by similar means, potentially through activation of shared immune pathways or energetic tradeoffs. Models predicting future ranges of both virus transmission and Wolbachia's efficacy following field release may wish to consider the effects these microbes have on host survival.}, } @article {pmid34292377, year = {2021}, author = {Manoj, RRS and Latrofa, MS and Mendoza-Roldan, JA and Otranto, D}, title = {Molecular detection of Wolbachia endosymbiont in reptiles and their ectoparasites.}, journal = {Parasitology research}, volume = {120}, number = {9}, pages = {3255-3261}, pmid = {34292377}, issn = {1432-1955}, mesh = {Animals ; *Ixodes/microbiology ; *Mites/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Reptiles/*microbiology/parasitology ; *Wolbachia/genetics/isolation & purification ; }, abstract = {Wolbachia, a maternally transmitted Gram-negative endosymbiont of onchocercid nematodes and arthropods, has a role in the biology of their host; thus it has been exploited for the filariasis treatment in humans. To assess the presence and prevalence of this endosymbiont in reptiles and their ectoparasites, blood and tail tissue as well as ticks and mites collected from them were molecularly screened for Wolbachia DNA using two sets of primers targeting partial 16S rRNA and Wolbachia surface protein (wsp) genes. Positive samples were screened for the partial 12S rRNA and cytochrome c oxidase subunit 1 (cox1) genes for filarioids. Of the different species of lizards (Podarcis siculus, Podarcis muralis and Lacerta bilineata) and snakes (Elaphe quatuorlineata and Boa constrictor constrictor) screened from three collection sites, only P. siculus scored positive for Wolbachia 16S rRNA. Among ectoparasites collected from reptiles (Ixodes ricinus ticks and Neotrombicula autumnalis, Ophionyssus sauracum and Ophionyssus natricis mites), I. ricinus (n = 4; 2.8%; 95% CI, 0.9-7) from P. siculus, N. autumnalis (n = 2 each; 2.8%; 95% CI, 0.9-6.5) from P. siculus and P. muralis and O. natricis (n = 1; 14.3%; 95% CI, 0.7-55.4) from Boa constrictor constrictor scored positive for Wolbachia DNA. None of the positive Wolbachia samples scored positive for filarioids. This represents the first report of Wolbachia in reptilian hosts and their ectoparasites, which follows a single identification in the intestinal cells of a filarioid associated with a gecko. This data could contribute to better understand the reptile filarioid-Wolbachia association and to unveil the evolutionary pattern of Wolbachia in its filarial host.}, } @article {pmid34288947, year = {2021}, author = {Tyagi, K and Tyagi, I and Kumar, V}, title = {Interspecific variation and functional traits of the gut microbiome in spiders from the wild: The largest effort so far.}, journal = {PloS one}, volume = {16}, number = {7}, pages = {e0251790}, pmid = {34288947}, issn = {1932-6203}, mesh = {Animals ; Ecosystem ; *Gastrointestinal Microbiome ; Species Specificity ; Spiders/*microbiology ; Symbiosis ; }, abstract = {Spiders being one of the most diverse group in phylum arthropod are of great importance due to their role as predators, silk producer, and in medicinal applications. Spiders in prey-predator relationships play a crucial role in balancing the food-chain of any ecosystem; therefore it is essential to characterize the gut microbiota of spiders collected from natural environments. In the present work, the largest effort so far has been made to characterize the gut microbiota of 35 spider species belonging to four different families using 16S amplicon targeting sequencing. Further, we compared the gut microbiota composition including endosymbiont abundance in spider species collected from different geographical locations. The results obtained revealed the presence of genera like Acinetobacter (15%), V7clade (9%), Wolbachia (8%), Pseudomonas (5%), Bacillus (6%). Although comparative analysis revealed that the gut bacterial composition in all the spider families has a similar pattern, in terms of community richness and evenness. The bacterial diversity in the spider family, Lycosidae are more diverse than in Salticidae, Tetragnathidae and Araneidae. Furthermore, it was observed that the abundance of endosymbiont genera, i.e. Wolbachia and Rickettsia, leads to shift in the abundance of other bacterial taxa and may cause sexual alterations in spider species. Moreover, predicted functional analysis based on PICRUSt2 reveals that gut microbiota of spider species were involved in functions like metabolism of carbohydrates, cofactors and vitamins, amino acids; biosynthesis of organic compounds, fatty acids, lipids etc. Based on the results obtained, it can be said that different locations do not correlate with community composition of gut microbiota in spider species collected from natural environments.}, } @article {pmid34282149, year = {2021}, author = {Wang, GH and Gamez, S and Raban, RR and Marshall, JM and Alphey, L and Li, M and Rasgon, JL and Akbari, OS}, title = {Combating mosquito-borne diseases using genetic control technologies.}, journal = {Nature communications}, volume = {12}, number = {1}, pages = {4388}, pmid = {34282149}, issn = {2041-1723}, support = {R01 AI151004/AI/NIAID NIH HHS/United States ; R01 AI116636/AI/NIAID NIH HHS/United States ; R21 AI149161/AI/NIAID NIH HHS/United States ; DP2 AI152071/AI/NIAID NIH HHS/United States ; BBS/E/I/00007033/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Animals, Genetically Modified ; CRISPR-Cas Systems ; Female ; Humans ; *Insecticides ; Malaria/prevention & control/transmission ; Male ; Mosquito Control/*methods ; Mosquito Vectors ; Nucleic Acid Amplification Techniques/*methods ; Pest Control, Biological ; Vector Borne Diseases/*genetics ; Wolbachia/genetics ; }, abstract = {Mosquito-borne diseases, such as dengue and malaria, pose significant global health burdens. Unfortunately, current control methods based on insecticides and environmental maintenance have fallen short of eliminating the disease burden. Scalable, deployable, genetic-based solutions are sought to reduce the transmission risk of these diseases. Pathogen-blocking Wolbachia bacteria, or genome engineering-based mosquito control strategies including gene drives have been developed to address these problems, both requiring the release of modified mosquitoes into the environment. Here, we review the latest developments, notable similarities, and critical distinctions between these promising technologies and discuss their future applications for mosquito-borne disease control.}, } @article {pmid34276627, year = {2021}, author = {Zhu, DH and Su, CY and Yang, XH and Abe, Y}, title = {A Case of Intragenic Recombination Dramatically Impacting the Phage WO Genetic Diversity in Gall Wasps.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {694115}, pmid = {34276627}, issn = {1664-302X}, abstract = {The phage WO was characterized in Wolbachia, a strictly intracellular bacterium causing several reproductive alterations in its arthropod hosts. This study aimed to screen the presence of Wolbachia and phage WO in 15 gall wasp species from six provinces of southern China to investigate their diversity and prevalence patterns. A high incidence of Wolbachia infection was determined in the gall wasp species, with an infection rate of 86.7% (13/15). Moreover, seven species had double or multiple infections. All Wolbachia-infected gall wasp species were found to harbor phage WO. The gall wasp species infected with a single Wolbachia strain were found to harbor a single phage WO type. On the contrary, almost all species with double or multiple Wolbachia infections harbored a high level of phage WO diversity (ranging from three to 27 types). Six horizontal transfer events of phage WO in Wolbachia were found to be associated with gall wasps, which shared identical orf7 sequences among their respective accomplices. The transfer potentially took place through gall inducers and associated inquilines infected with or without Wolbachia. Furthermore, 10 putative recombination events were identified from Andricus hakonensis and Andricus sp2, which harbored multiple phage WO types, suggesting that intragenic recombination was the important evolutionary force, which effectively promoted the high level of phage WO diversity associated with gall wasps.}, } @article {pmid34276617, year = {2021}, author = {Huang, J and Dai, Z and Zheng, Z and da Silvia, PA and Kumagai, L and Xiang, Q and Chen, J and Deng, X}, title = {Bacteriomic Analyses of Asian Citrus Psyllid and Citrus Samples Infected With "Candidatus Liberibacter asiaticus" in Southern California and Huanglongbing Management Implications.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {683481}, pmid = {34276617}, issn = {1664-302X}, abstract = {Citrus Huanglongbing (HLB; yellow shoot disease) is associated with an unculturable α-proteobacterium "Candidatus Liberibacter asiaticus" (CLas). HLB was found in southern California in 2012, and the current management strategy is based on suppression of the Asian citrus psyllid (Diaphorina citri) that transmits CLas and removal of confirmed CLas-positive trees. Little is known about Asian citrus psyllid-associated bacteria and citrus-associated bacteria in the HLB system. Such information is important in HLB management, particularly for accurate detection of CLas. Recent advancements in next-generation sequencing technology provide new opportunities to study HLB through genomic DNA sequence analyses (metagenomics). In this study, HLB-related bacteria in Asian citrus psyllid and citrus (represented by leaf midrib tissues) samples from southern California were analyzed. A metagenomic pipeline was developed to serve as a prototype for future bacteriomic research. This pipeline included steps of next-generation sequencing in Illumina platform, de novo assembly of Illumina reads, sequence classification using the Kaiju tool, acquisition of bacterial draft genome sequences, and taxonomic validation and diversity evaluation using average nucleotide identity. The identified bacteria in Asian citrus psyllids and citrus together included Bradyrhizobium, Buchnera, Burkholderia, "Candidatus Profftella armature," "Candidatus Carsonella ruddii," CLas, Mesorhizobium, Paraburkholderia, Pseudomonas, and Wolbachia. The whole genome of a CLas strain recently found in San Bernardino County was sequenced and classified into prophage typing group 1 (PTG-1), one of the five known CLas groups in California. Based on sequence similarity, Bradyrhizobium and Mesorhizobium were identified as possible source that could interfere with CLas detection using the 16S rRNA gene-based PCR commonly used for HLB diagnosis, particularly at low or zero CLas titer situation.}, } @article {pmid34275443, year = {2021}, author = {Liew, C and Soh, LT and Chen, I and Ng, LC}, title = {Public sentiments towards the use of Wolbachia-Aedes technology in Singapore.}, journal = {BMC public health}, volume = {21}, number = {1}, pages = {1417}, pmid = {34275443}, issn = {1471-2458}, mesh = {*Aedes ; Animals ; *Dengue ; Humans ; Male ; Mosquito Vectors ; Singapore ; Technology ; *Wolbachia ; }, abstract = {BACKGROUND: Wolbachia technology is a novel vector control approach that can reduce mosquito populations and the risk of mosquito-borne diseases, which has recently gained popularity amongst countries. In 2016, Singapore embarked on a multi-phased field study named Project Wolbachia - Singapore, to evaluate the use of Wolbachia technology as an Aedes aegypti mosquito population suppression tool to fight dengue. Due to the novelty of this technology in Singapore, this study aims to understand the public's acceptance and sentiments towards the use of Wolbachia technology.

METHODS: Several public sentiment survey approaches - including online, face-to-face in the streets, as well as door-to-door household surveys - were conducted.

RESULTS: The surveys conducted prior to the first field releases and implementation of the project revealed high support for the use of Wolbachia technology in Singapore. A household perception survey conducted in the interim of the first project phase was encouraging, with the majority of the respondents being aware of the project and having no concerns with the release of male Wolbachia-carrying Aedes aegypti (Wolbachia-Aedes) mosquitoes in their neighbourhood.

CONCLUSIONS: The study reveal high support for the use of Wolbachia technology in Singapore and also provided invaluable insights that were used in the development of a public communications and engagement framework model, which thus helped to guide these elements in the subsequent phases and expansion of the project.}, } @article {pmid34273392, year = {2021}, author = {Gangwar, M and Jha, R and Goyal, M and Srivastava, M}, title = {Biochemical characterization of Recombinase A from Wolbachia endosymbiont of filarial nematode Brugia malayi (wBmRecA).}, journal = {International journal for parasitology}, volume = {51}, number = {10}, pages = {841-853}, doi = {10.1016/j.ijpara.2021.02.007}, pmid = {34273392}, issn = {1879-0135}, mesh = {Animals ; *Brugia malayi ; *Elephantiasis, Filarial ; Female ; Humans ; Microfilariae ; Rec A Recombinases/antagonists & inhibitors/chemistry/*metabolism ; *Wolbachia ; }, abstract = {Lymphatic filariasis is a debilitating disease that affects over 890 million people in 49 countries. A lack of vaccines, non-availability of adulticidal drugs, the threat of emerging drug resistance against available chemotherapeutics and an incomplete understanding of the immunobiology of the disease have sustained the problem. Characterization of Wolbachia proteins, the bacterial endosymbiont which helps in the growth and development of filarial worms, regulates fecundity in female worms and mediates immunopathogenesis of Lymphatic Filariasis, is an important approach to gain insights into the immunopathogenesis of the disease. In this study, we carried out extensive biochemical characterization of Recombinase A from Wolbachia of the filarial nematode Brugia malayi (wBmRecA) using an Electrophoretic Mobility Shift Assay, an ATP binding and hydrolysis assay, DNA strand exchange reactions, DAPI displacement assay and confocal microscopy, and evaluated anti-filarial activity of RecA inhibitors. Confocal studies showed that wBmRecA was expressed and localised within B. malayi microfilariae (Mf) and uteri and lateral chord of adult females. Recombinant wBmRecA was biochemically active and showed intrinsic binding capacity towards both single-stranded DNA and double-stranded DNA that were enhanced by ATP, suggesting ATP-induced cooperativity. wBmRecA promoted ATP hydrolysis and DNA strand exchange reactions in a concentration-dependent manner, and its binding to DNA was sensitive to temperature, pH and salt concentration. Importantly, the anti-parasitic drug Suramin, and Phthalocyanine tetrasulfonate (PcTs)-based inhibitors Fe-PcTs and 3,4-Cu-PcTs, inhibited wBmRecA activity and affected the motility and viability of Mf. The addition of Doxycycline further enhanced microfilaricidal activity of wBmRecA, suggesting potential synergism. Taken together, the omnipresence of wBmRecA in B. malayi life stages and the potent microfilaricidal activity of RecA inhibitors suggest an important role of wBmRecA in filarial pathogenesis.}, } @article {pmid34273308, year = {2021}, author = {Ross, PA}, title = {Designing effective Wolbachia release programs for mosquito and arbovirus control.}, journal = {Acta tropica}, volume = {222}, number = {}, pages = {106045}, doi = {10.1016/j.actatropica.2021.106045}, pmid = {34273308}, issn = {1873-6254}, mesh = {*Aedes/microbiology ; Animals ; *Arboviruses ; Female ; Male ; Mosquito Control/*methods ; Mosquito Vectors/microbiology ; *Wolbachia ; }, abstract = {Mosquitoes carrying endosymbiotic bacteria called Wolbachia are being released in mosquito and arbovirus control programs around the world through two main approaches: population suppression and population replacement. Open field releases of Wolbachia-infected male mosquitoes have achieved over 95% population suppression by reducing the fertility of wild mosquito populations. The replacement of populations with Wolbachia-infected females is self-sustaining and can greatly reduce local dengue transmission by reducing the vector competence of mosquito populations. Despite many successful interventions, significant questions and challenges lie ahead. Wolbachia, viruses and their mosquito hosts can evolve, leading to uncertainty around the long-term effectiveness of a given Wolbachia strain, while few ecological impacts of Wolbachia releases have been explored. Wolbachia strains are diverse and the choice of strain to release should be made carefully, taking environmental conditions and the release objective into account. Mosquito quality control, thoughtful community awareness programs and long-term monitoring of populations are essential for all types of Wolbachia intervention. Releases of Wolbachia-infected mosquitoes show great promise, but existing control measures remain an important way to reduce the burden of mosquito-borne disease.}, } @article {pmid34272503, year = {2021}, author = {Russell, A and Borrelli, S and Fontana, R and Laricchiuta, J and Pascar, J and Becking, T and Giraud, I and Cordaux, R and Chandler, CH}, title = {Evolutionary transition to XY sex chromosomes associated with Y-linked duplication of a male hormone gene in a terrestrial isopod.}, journal = {Heredity}, volume = {127}, number = {3}, pages = {266-277}, pmid = {34272503}, issn = {1365-2540}, mesh = {Animals ; Evolution, Molecular ; Female ; Genome ; Hormones ; Humans ; *Isopoda/genetics ; Male ; Sex Chromosomes/genetics ; Sex Determination Processes/genetics ; }, abstract = {Sex chromosomes are highly variable in some taxonomic groups, but the evolutionary mechanisms underlying this diversity are not well understood. In terrestrial isopod crustaceans, evolutionary turnovers in sex chromosomes are frequent, possibly caused by Wolbachia, a vertically-transmitted endosymbiont causing male-to-female sex reversal. Here, we use surgical manipulations and genetic crosses, plus genome sequencing, to examine sex chromosomes in the terrestrial isopod Trachelipus rathkei. Although an earlier cytogenetics study suggested a ZZ/ZW sex chromosome system in this species, we surprisingly find multiple lines of evidence that in our study population, sex is determined by an XX/XY system. Consistent with a recent evolutionary origin for this XX/XY system, the putative male-specific region of the genome is small. The genome shows evidence of Y-linked duplications of the gene encoding the androgenic gland hormone, a major component of male sexual differentiation in isopods. Our analyses also uncover sequences horizontally acquired from past Wolbachia infections, consistent with the hypothesis that Wolbachia may have interfered with the evolution of sex determination in T. rathkei. Overall, these results provide evidence for the co-occurrence of multiple sex chromosome systems within T. rathkei, further highlighting the relevance of terrestrial isopods as models for the study of sex chromosome evolution.}, } @article {pmid34268130, year = {2021}, author = {Kee, SL and Tan, MJT}, title = {Friend, Not Foe: Unveiling Vector-Bacteria Symbiosis and Its Utility as an Arboviral Intervention Strategy in the Philippines.}, journal = {Frontiers in cellular and infection microbiology}, volume = {11}, number = {}, pages = {650277}, pmid = {34268130}, issn = {2235-2988}, mesh = {*Aedes ; Animals ; Friends ; Humans ; Mosquito Vectors ; Philippines ; Symbiosis ; *Wolbachia ; }, } @article {pmid34264019, year = {2022}, author = {Zhu, DT and Rao, Q and Zou, C and Ban, FX and Zhao, JJ and Liu, SS}, title = {Genomic and transcriptomic analyses reveal metabolic complementarity between whiteflies and their symbionts.}, journal = {Insect science}, volume = {29}, number = {2}, pages = {539-549}, doi = {10.1111/1744-7917.12943}, pmid = {34264019}, issn = {1744-7917}, mesh = {Animals ; Genomics ; *Halomonadaceae ; *Hemiptera/microbiology ; Symbiosis/genetics ; Transcriptome ; }, abstract = {Nutritional mutualism between insects and symbiotic bacteria is widespread. The various sap-feeding whitefly species within the Bemisia tabaci complex associate with the same obligate symbiont (Portiera) and multiple secondary symbionts. It is often assumed that some of the symbionts residing in the whiteflies play crucial roles in the nutritional physiology of their insect hosts. Although effort has been made to understand the functions of the whitefly symbionts, the metabolic complementarity offered by these symbionts to the hosts is not yet well understood. We examined two secondary symbionts, Arsenophonus and Wolbachia, in two species of the B. tabaci whitefly complex, provisionally named as Asia II 3 and China 1. Genomic sequence analyses revealed that Arsenophonus and Wolbachia retained genes responsible for the biosynthesis of B vitamins. We then conducted transcriptomic surveys of the bacteriomes in these two species of whiteflies together with that in another species named MED of this whitefly complex previously reported. The analyses indicated that several key genes in B vitamin syntheses from the three whitefly species were identical. Our findings suggest that, similar to another secondary symbiont Hamiltonella, Arsenophonus and Wolbachia function in the nutrient provision of host whiteflies. Although phylogenetically distant species of symbionts are associated with their respective hosts, they have evolved and retained similar functions in biosynthesis of some B vitamins. Such metabolic complementarity between whiteflies and symbionts represents an important feature of their coevolution.}, } @article {pmid34253453, year = {2022}, author = {Massey, JH and Newton, ILG}, title = {Diversity and function of arthropod endosymbiont toxins.}, journal = {Trends in microbiology}, volume = {30}, number = {2}, pages = {185-198}, pmid = {34253453}, issn = {1878-4380}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Arthropods ; Male ; Phylogeny ; *Rickettsia ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Bacterial endosymbionts induce dramatic phenotypes in their arthropod hosts, including cytoplasmic incompatibility, feminization, parthenogenesis, male killing, parasitoid defense, and pathogen blocking. The molecular mechanisms underlying these effects remain largely unknown but recent evidence suggests that protein toxins secreted by the endosymbionts play a role. Here, we describe the diversity and function of endosymbiont proteins with homology to known bacterial toxins. We focus on maternally transmitted endosymbionts belonging to the Wolbachia, Rickettsia, Arsenophonus, Hamiltonella, Spiroplasma, and Cardinium genera because of their ability to induce the above phenotypes. We identify at least 16 distinct toxin families with diverse enzymatic activities, including AMPylases, nucleases, proteases, and glycosyltransferases. Notably, several annotated toxins contain domains with homology to eukaryotic proteins, suggesting that arthropod endosymbionts mimic host biochemistry to manipulate host physiology, similar to bacterial pathogens.}, } @article {pmid34252106, year = {2021}, author = {Pinto, SB and Riback, TIS and Sylvestre, G and Costa, G and Peixoto, J and Dias, FBS and Tanamas, SK and Simmons, CP and Dufault, SM and Ryan, PA and O'Neill, SL and Muzzi, FC and Kutcher, S and Montgomery, J and Green, BR and Smithyman, R and Eppinghaus, A and Saraceni, V and Durovni, B and Anders, KL and Moreira, LA}, title = {Effectiveness of Wolbachia-infected mosquito deployments in reducing the incidence of dengue and other Aedes-borne diseases in Niterói, Brazil: A quasi-experimental study.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {7}, pages = {e0009556}, pmid = {34252106}, issn = {1935-2735}, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Brazil/epidemiology ; Chikungunya Fever/epidemiology/*transmission/virology ; Chikungunya virus/physiology ; Dengue/epidemiology/*transmission/virology ; Dengue Virus/physiology ; Female ; Humans ; Incidence ; Male ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/physiology/virology ; Wolbachia/*physiology ; Zika Virus/physiology ; Zika Virus Infection/epidemiology/*transmission/virology ; }, abstract = {BACKGROUND: The introduction of the bacterium Wolbachia (wMel strain) into Aedes aegypti mosquitoes reduces their capacity to transmit dengue and other arboviruses. Evidence of a reduction in dengue case incidence following field releases of wMel-infected Ae. aegypti has been reported previously from a cluster randomised controlled trial in Indonesia, and quasi-experimental studies in Indonesia and northern Australia.

Following pilot releases in 2015-2016 and a period of intensive community engagement, deployments of adult wMel-infected Ae. aegypti mosquitoes were conducted in Niterói, Brazil during 2017-2019. Deployments were phased across four release zones, with a total area of 83 km2 and a residential population of approximately 373,000. A quasi-experimental design was used to evaluate the effectiveness of wMel deployments in reducing dengue, chikungunya and Zika incidence. An untreated control zone was pre-defined, which was comparable to the intervention area in historical dengue trends. The wMel intervention effect was estimated by controlled interrupted time series analysis of monthly dengue, chikungunya and Zika case notifications to the public health surveillance system before, during and after releases, from release zones and the control zone. Three years after commencement of releases, wMel introgression into local Ae. aegypti populations was heterogeneous throughout Niterói, reaching a high prevalence (>80%) in the earliest release zone, and more moderate levels (prevalence 40-70%) elsewhere. Despite this spatial heterogeneity in entomological outcomes, the wMel intervention was associated with a 69% reduction in dengue incidence (95% confidence interval 54%, 79%), a 56% reduction in chikungunya incidence (95%CI 16%, 77%) and a 37% reduction in Zika incidence (95%CI 1%, 60%), in the aggregate release area compared with the pre-defined control area. This significant intervention effect on dengue was replicated across all four release zones, and in three of four zones for chikungunya, though not in individual release zones for Zika.

CONCLUSIONS/SIGNIFICANCE: We demonstrate that wMel Wolbachia can be successfully introgressed into Ae. aegypti populations in a large and complex urban setting, and that a significant public health benefit from reduced incidence of Aedes-borne disease accrues even where the prevalence of wMel in local mosquito populations is moderate and spatially heterogeneous. These findings are consistent with the results of randomised and non-randomised field trials in Indonesia and northern Australia, and are supportive of the Wolbachia biocontrol method as a multivalent intervention against dengue, chikungunya and Zika.}, } @article {pmid34252087, year = {2021}, author = {Noroy, C and Meyer, DF}, title = {The super repertoire of type IV effectors in the pangenome of Ehrlichia spp. provides insights into host-specificity and pathogenesis.}, journal = {PLoS computational biology}, volume = {17}, number = {7}, pages = {e1008788}, pmid = {34252087}, issn = {1553-7358}, mesh = {Animals ; Bacterial Proteins ; Computational Biology ; *Ehrlichia/genetics/pathogenicity ; Ehrlichiosis/microbiology ; Genome, Bacterial/*genetics ; Host Specificity/*genetics ; Humans ; Type IV Secretion Systems/*genetics ; Virulence/*genetics ; }, abstract = {The identification of bacterial effectors is essential to understand how obligatory intracellular bacteria such as Ehrlichia spp. manipulate the host cell for survival and replication. Infection of mammals-including humans-by the intracellular pathogenic bacteria Ehrlichia spp. depends largely on the injection of virulence proteins that hijack host cell processes. Several hypothetical virulence proteins have been identified in Ehrlichia spp., but one so far has been experimentally shown to translocate into host cells via the type IV secretion system. However, the current challenge is to identify most of the type IV effectors (T4Es) to fully understand their role in Ehrlichia spp. virulence and host adaptation. Here, we predict the T4E repertoires of four sequenced Ehrlichia spp. and four other Anaplasmataceae as comparative models (pathogenic Anaplasma spp. and Wolbachia endosymbiont) using previously developed S4TE 2.0 software. This analysis identified 579 predicted T4Es (228 pT4Es for Ehrlichia spp. only). The effector repertoires of Ehrlichia spp. overlapped, thereby defining a conserved core effectome of 92 predicted effectors shared by all strains. In addition, 69 species-specific T4Es were predicted with non-canonical GC% mostly in gene sparse regions of the genomes and we observed a bias in pT4Es according to host-specificity. We also identified new protein domain combinations, suggesting novel effector functions. This work presenting the predicted effector collection of Ehrlichia spp. can serve as a guide for future functional characterisation of effectors and design of alternative control strategies against these bacteria.}, } @article {pmid34249780, year = {2021}, author = {Reyes, JIL and Suzuki, Y and Carvajal, T and Muñoz, MNM and Watanabe, K}, title = {Intracellular Interactions Between Arboviruses and Wolbachia in Aedes aegypti.}, journal = {Frontiers in cellular and infection microbiology}, volume = {11}, number = {}, pages = {690087}, pmid = {34249780}, issn = {2235-2988}, mesh = {*Aedes ; Animals ; *Arboviruses ; Humans ; Mosquito Vectors ; *Wolbachia ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {Aedes aegypti is inherently susceptible to arboviruses. The geographical expansion of this vector host species has led to the persistence of Dengue, Zika, and Chikungunya human infections. These viruses take advantage of the mosquito's cell to create an environment conducive for their growth. Arboviral infection triggers transcriptomic and protein dysregulation in Ae. aegypti and in effect, host antiviral mechanisms are compromised. Currently, there are no existing vaccines able to protect human hosts from these infections and thus, vector control strategies such as Wolbachia mass release program is regarded as a viable option. Considerable evidence demonstrates how the presence of Wolbachia interferes with arboviruses by decreasing host cytoskeletal proteins and lipids essential for arboviral infection. Also, Wolbachia strengthens host immunity, cellular regeneration and causes the expression of microRNAs which could potentially be involved in virus inhibition. However, variation in the magnitude of Wolbachia's pathogen blocking effect that is not due to the endosymbiont's density has been recently reported. Furthermore, the cellular mechanisms involved in this phenotype differs depending on Wolbachia strain and host species. This prompts the need to explore the cellular interactions between Ae. aegypti-arboviruses-Wolbachia and how different Wolbachia strains overall affect the mosquito's cell. Understanding what happens at the cellular and molecular level will provide evidence on the sustainability of Wolbachia vector control.}, } @article {pmid34247634, year = {2021}, author = {Endersby-Harshman, NM and Ali, A and Alhumrani, B and Alkuriji, MA and Al-Fageeh, MB and Al-Malik, A and Alsuabeyl, MS and Elfekih, S and Hoffmann, AA}, title = {Voltage-sensitive sodium channel (Vssc) mutations associated with pyrethroid insecticide resistance in Aedes aegypti (L.) from two districts of Jeddah, Kingdom of Saudi Arabia: baseline information for a Wolbachia release program.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {361}, pmid = {34247634}, issn = {1756-3305}, mesh = {Aedes/*drug effects/*genetics ; Animals ; Biological Assay/methods/statistics & numerical data ; Dengue/prevention & control/transmission ; Female ; Insecticide Resistance/*genetics ; Insecticides/*pharmacology ; Mosquito Control/methods/statistics & numerical data ; *Mutation ; Pyrethrins/*pharmacology ; Saudi Arabia ; Sodium Channels/*genetics ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Dengue suppression often relies on control of the mosquito vector, Aedes aegypti, through applications of insecticides of which the pyrethroid group has played a dominant role. Insecticide resistance is prevalent in Ae. aegypti around the world, and the resulting reduction of insecticide efficacy is likely to exacerbate the impact of dengue. Dengue has been a public health problem in Saudi Arabia, particularly in Jeddah, since its discovery there in the 1990s, and insecticide use for vector control is widespread throughout the city. An alternative approach to insecticide use, based on blocking dengue transmission in mosquitoes by the endosymbiont Wolbachia, is being trialed in Jeddah following the success of this approach in Australia and Malaysia. Knowledge of insecticide resistance status of mosquito populations in Jeddah is a prerequisite for establishing a Wolbachia-based dengue control program as releases of Wolbachia mosquitoes succeed when resistance status of the release population is similar to that of the wild population.

METHODS: WHO resistance bioassays of mosquitoes with deltamethrin, permethrin and DDT were used in conjunction with TaqMan[®] SNP Genotyping Assays to characterize mutation profiles of Ae. aegypti.

RESULTS: Screening of the voltage-sensitive sodium channel (Vssc), the pyrethroid target site, revealed mutations at codons 989, 1016 and 1534 in Ae. aegypti from two districts of Jeddah. The triple mutant homozygote (1016G/1534C/989P) was confirmed from Al Safa and Al Rawabi. Bioassays with pyrethroids (Type I and II) and DDT showed that mosquitoes were resistant to each of these compounds based on WHO definitions. An association between Vssc mutations and resistance was established for the Type II pyrethroid, deltamethrin, with one genotype (989P/1016G/1534F) conferring a survival advantage over two others (989S/1016V/1534C and the triple heterozygote). An indication of synergism of Type I pyrethroid activity with piperonyl butoxide suggests that detoxification by cytochrome P450s accounts for some of the pyrethroid resistance response in Ae. aegypti populations from Jeddah.

CONCLUSIONS: The results provide a baseline for monitoring and management of resistance as well as knowledge of Vssc genotype frequencies required in Wolbachia release populations to ensure homogeneity with the target field population. Vssc mutation haplotypes observed show some similarity with those from Ae. aegypti in southeast Asia and the Indo-Pacific, but the presence of the triple mutant haplotype in three genotypes indicates that the species in this region may have a unique population history.}, } @article {pmid34233062, year = {2021}, author = {Duplouy, A and Nair, A and Nyman, T and van Nouhuys, S}, title = {Long-term spatiotemporal genetic structure of an accidental parasitoid introduction, and local changes in prevalence of its associated Wolbachia symbiont.}, journal = {Molecular ecology}, volume = {30}, number = {18}, pages = {4368-4380}, doi = {10.1111/mec.16065}, pmid = {34233062}, issn = {1365-294X}, mesh = {Animals ; *Butterflies/genetics ; Humans ; Microsatellite Repeats ; Prevalence ; *Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {Population bottlenecks associated with founder events strongly impact the establishment and genetic makeup of populations. In addition to their genotype, founding individuals also bring along parasites, as well as symbionts that can manipulate the phenotype of their host, affecting the host population establishment, dynamics and evolution. Thus, to understand introduction, invasion, and spread, we should identify the roles played by accompanying symbionts. In 1991, the parasitoid wasp, Hyposoter horticola, and its associated hyperparasitoid were accidentally introduced from the main Åland islands, Finland, to an isolated island in the archipelago, along with their host, the Glanville fritillary butterfly. Though the receiving island was unoccupied, the butterfly was present on some of the small islands in the vicinity. The three introduced species have persisted locally ever since. A strain of the endosymbiotic bacterium Wolbachia has an intermediate prevalence in the parasitoid H. horticola across the main Åland population. The infection increases its susceptibility of to hyperparasitism. We investigated the establishment and spread of the parasitoid, along with patterns of prevalence of its symbiont using 323 specimens collected between 1992 and 2013, from five localities across Åland, including the source and introduced populations. Using 14 microsatellites and one mitochondrial marker, we suggest that the relatively diverse founding population and occasional migration between islands might have facilitated the persistence of all isolated populations, despite multiple local population crashes. We also show that where the hyperparasitoid is absent, and thus selection against infected wasp genotypes is relaxed, there is near-fixation of Wolbachia.}, } @article {pmid34229954, year = {2021}, author = {Johnston, KL and Hong, WD and Turner, JD and O'Neill, PM and Ward, SA and Taylor, MJ}, title = {Anti-Wolbachia drugs for filariasis.}, journal = {Trends in parasitology}, volume = {37}, number = {12}, pages = {1068-1081}, doi = {10.1016/j.pt.2021.06.004}, pmid = {34229954}, issn = {1471-5007}, support = {NC/M00175X/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; /MRC_/Medical Research Council/United Kingdom ; }, mesh = {Anti-Bacterial Agents/pharmacology/therapeutic use ; Drug Discovery ; *Elephantiasis, Filarial/drug therapy ; Humans ; *Nematode Infections/drug therapy ; *Onchocerciasis/drug therapy ; *Wolbachia ; }, abstract = {The mutualistic association between Wolbachia endosymbionts and their filarial nematode hosts has been exploited as a validated drug target delivering macrofilaricidal outcomes. Limitations of existing antibiotics to scale-up have driven the search for new drugs, which are effective in shorter regimens of 7 days or less. Here, we review the last 14 years of anti-Wolbachia drug discovery by the anti-Wolbachia (A·WOL) consortium, which has screened more than two million compounds, delivering thousands of hit compounds. Refined screening models integrated with robust pharmacokinetic/pharmacodynamic (PK/PD) driven optimisation and selection strategies have delivered the first two drug candidates specifically designed to target Wolbachia. AWZ1066S and ABBV-4083 are currently progressing through clinical trials with the aim of delivering safe and effective macrofilaricides to support the elimination of onchocerciasis and lymphatic filariasis.}, } @article {pmid34221442, year = {2021}, author = {Chan, PK and Hawley, JR and Lappin, MR}, title = {Evaluation of the role of Babesia species and Cytauxzoon felis in feline anemia cases in Colorado, USA.}, journal = {JFMS open reports}, volume = {7}, number = {1}, pages = {20551169211024967}, pmid = {34221442}, issn = {2055-1169}, abstract = {OBJECTIVES: The aim of this study was to evaluate the blood of cats in Colorado, USA, with suspected infectious causes of anemia for the presence of Babesia species and Cytauxzoon felis DNA. Results of PCR testing for other common vector-borne diseases potentially associated with anemia are also reported.

METHODS: Samples from 101 cats were tested using a PCR assay that coamplified the DNA of C felis and Babesia species mitochondrial DNA. PCR testing for DNA of hemoplasmas, Bartonella species, Ehrlichia species, Anaplasma species, Neorickettsia risticii and Wolbachia genera was also performed if not carried out previously.

RESULTS: Twenty-two cats (21.8%) were positive for DNA of an infectious agent. DNA from hemoplasma species were amplified from 14 cats (13.9%). Bartonella species DNA was amplified from four cats (4%) and Ehrlichia canis, Anaplasma platys, Anaplasma phagocytophilum and Wolbachia genera DNA were amplified from one cat each. Babesia species and C felis mitochondrial DNA were not amplified from any sample.

CONCLUSIONS AND RELEVANCE: Based on the results of this study, it does not appear that Babesia species or C felis are clinically relevant in anemic cats in Colorado, USA. For C felis, this suggests that the vector Amblyomma americanum is still uncommon in this geographic area.}, } @article {pmid34217365, year = {2021}, author = {Lejal, E and Chiquet, J and Aubert, J and Robin, S and Estrada-Peña, A and Rue, O and Midoux, C and Mariadassou, M and Bailly, X and Cougoul, A and Gasqui, P and Cosson, JF and Chalvet-Monfray, K and Vayssier-Taussat, M and Pollet, T}, title = {Temporal patterns in Ixodes ricinus microbial communities: an insight into tick-borne microbe interactions.}, journal = {Microbiome}, volume = {9}, number = {1}, pages = {153}, pmid = {34217365}, issn = {2049-2618}, mesh = {Animals ; *Borrelia ; Humans ; *Ixodes ; Microbial Interactions ; *Microbiota/genetics ; *Rickettsia/genetics ; }, abstract = {BACKGROUND: Ticks transmit pathogens of medical and veterinary importance and are an increasing threat to human and animal health. Assessing disease risk and developing new control strategies requires identifying members of the tick-borne microbiota as well as their temporal dynamics and interactions.

METHODS: Using high-throughput sequencing, we studied the Ixodes ricinus microbiota and its temporal dynamics. 371 nymphs were monthly collected during three consecutive years in a peri-urban forest. After a Poisson lognormal model was adjusted to our data set, a principal component analysis, sparse network reconstruction, and differential analysis allowed us to assess seasonal and monthly variability of I. ricinus microbiota and interactions within this community.

RESULTS: Around 75% of the detected sequences belonged to five genera known to be maternally inherited bacteria in arthropods and to potentially circulate in ticks: Candidatus Midichloria, Rickettsia, Spiroplasma, Arsenophonus and Wolbachia. The structure of the I. ricinus microbiota varied over time with interannual recurrence and seemed to be mainly driven by OTUs commonly found in the environment. Total network analysis revealed a majority of positive partial correlations. We identified strong relationships between OTUs belonging to Wolbachia and Arsenophonus, evidence for the presence of the parasitoid wasp Ixodiphagus hookeri in ticks. Other associations were observed between the tick symbiont Candidatus Midichloria and pathogens belonging to Rickettsia. Finally, more specific network analyses were performed on TBP-infected samples and suggested that the presence of pathogens belonging to the genera Borrelia, Anaplasma and Rickettsia may disrupt microbial interactions in I. ricinus.

CONCLUSIONS: We identified the I. ricinus microbiota and documented marked shifts in tick microbiota dynamics over time. Statistically, we showed strong relationships between the presence of specific pathogens and the structure of the I. ricinus microbiota. We detected close links between some tick symbionts and the potential presence of either pathogenic Rickettsia or a parasitoid in ticks. These new findings pave the way for the development of new strategies for the control of ticks and tick-borne diseases. Video abstract.}, } @article {pmid34208681, year = {2021}, author = {Ourry, M and Crosland, A and Lopez, V and Derocles, SAP and Mougel, C and Cortesero, AM and Poinsot, D}, title = {Influential Insider: Wolbachia, an Intracellular Symbiont, Manipulates Bacterial Diversity in Its Insect Host.}, journal = {Microorganisms}, volume = {9}, number = {6}, pages = {}, pmid = {34208681}, issn = {2076-2607}, support = {SE 0000282 MP-P10026 EB06//Institut National de la Recherche Agronomique/ ; }, abstract = {Facultative intracellular symbionts like the α-proteobacteria Wolbachia influence their insect host phenotype but little is known about how much they affect their host microbiota. Here, we quantified the impact of Wolbachia infection on the bacterial community of the cabbage root fly Delia radicum by comparing the microbiota of Wolbachia-free and infected adult flies of both sexes. We used high-throughput DNA sequencing (Illumina MiSeq, 16S rRNA, V5-V7 region) and performed a community and a network analysis. In both sexes, Wolbachia infection significantly decreased the diversity of D. radicum bacterial communities and modified their structure and composition by reducing abundance in some taxa but increasing it in others. Infection by Wolbachia was negatively correlated to 8 bacteria genera (Erwinia was the most impacted), and positively correlated to Providencia and Serratia. We suggest that Wolbachia might antagonize Erwinia for being entomopathogenic (and potentially intracellular), but would favor Providencia and Serratia because they might protect the host against chemical plant defenses. Although they might seem prisoners in a cell, endocellular symbionts can impact the whole microbiota of their host, hence its extended phenotype, which provides them with a way to interact with the outside world.}, } @article {pmid34197460, year = {2021}, author = {Kupritz, J and Martin, J and Fischer, K and Curtis, KC and Fauver, JR and Huang, Y and Choi, YJ and Beatty, WL and Mitreva, M and Fischer, PU}, title = {Isolation and characterization of a novel bacteriophage WO from Allonemobius socius crickets in Missouri.}, journal = {PloS one}, volume = {16}, number = {7}, pages = {e0250051}, pmid = {34197460}, issn = {1932-6203}, mesh = {Animals ; Bacteriophages/classification/*genetics/isolation & purification ; Capsid Proteins/genetics ; DNA, Bacterial/chemistry/metabolism ; DNA, Viral/chemistry/metabolism ; Female ; *Genome, Viral ; Gryllidae/*microbiology/virology ; Membrane Proteins/genetics ; Missouri ; Open Reading Frames/genetics ; Phylogeny ; Whole Genome Sequencing ; Wolbachia/genetics/isolation & purification/virology ; }, abstract = {Wolbachia are endosymbionts of numerous arthropod and some nematode species, are important for their development and if present can cause distinct phenotypes of their hosts. Prophage DNA has been frequently detected in Wolbachia, but particles of Wolbachia bacteriophages (phage WO) have been only occasionally isolated. Here, we report the characterization and isolation of a phage WO of the southern ground cricket, Allonemobius socius, and provided the first whole-genome sequence of phage WO from this arthropod family outside of Asia. We screened A. socius abdomen DNA extracts from a cricket population in eastern Missouri by quantitative PCR for Wolbachia surface protein and phage WO capsid protein and found a prevalence of 55% and 50%, respectively, with many crickets positive for both. Immunohistochemistry using antibodies against Wolbachia surface protein showed many Wolbachia clusters in the reproductive system of female crickets. Whole-genome sequencing using Oxford Nanopore MinION and Illumina technology allowed for the assembly of a high-quality, 55 kb phage genome containing 63 open reading frames (ORF) encoding for phage WO structural proteins and host lysis and transcriptional manipulation. Taxonomically important regions of the assembled phage genome were validated by Sanger sequencing of PCR amplicons. Analysis of the nucleotides sequences of the ORFs encoding the large terminase subunit (ORF2) and minor capsid (ORF7) frequently used for phage WO phylogenetics showed highest homology to phage WOAu of Drosophila simulans (94.46% identity) and WOCin2USA1 of the cherry fruit fly, Rhagoletis cingulata (99.33% identity), respectively. Transmission electron microscopy examination of cricket ovaries showed a high density of phage particles within Wolbachia cells. Isolation of phage WO revealed particles characterized by 40-62 nm diameter heads and up to 190 nm long tails. This study provides the first detailed description and genomic characterization of phage WO from North America that is easily accessible in a widely distributed cricket species.}, } @article {pmid34190587, year = {2021}, author = {Leitner, M and Bishop, C and Asgari, S}, title = {Transcriptional Response of Wolbachia to Dengue Virus Infection in Cells of the Mosquito Aedes aegypti.}, journal = {mSphere}, volume = {6}, number = {3}, pages = {e0043321}, pmid = {34190587}, issn = {2379-5042}, abstract = {Aedes aegypti transmits one of the most significant mosquito-borne viruses, dengue virus (DENV). The absence of effective vaccines and clinical treatments and the emergence of insecticide resistance in A. aegypti necessitate novel vector control strategies. A new approach uses the endosymbiotic bacterium Wolbachia pipientis to reduce the spread of arboviruses. However, the Wolbachia-mediated antiviral mechanism is not well understood. To shed light on this mechanism, we investigated an unexplored aspect of Wolbachia-virus-mosquito interaction. We used RNA sequencing to examine the transcriptional response of Wolbachia to DENV infection in A. aegypti Aag2 cells transinfected with the wAlbB strain of Wolbachia. Our results suggest that genes encoding an endoribonuclease (RNase HI), a regulator of sigma 70-dependent gene transcription (6S RNA), essential cellular, transmembrane, and stress response functions and primary type I and IV secretion systems were upregulated, while a number of transport and binding proteins of Wolbachia, ribosome structure, and elongation factor-associated genes were downregulated due to DENV infection. Furthermore, bacterial retrotransposon, transposable, and phage-related elements were found among the up- and downregulated genes. We show that Wolbachia elicits a transcriptional response to virus infection and identify differentially expressed Wolbachia genes mostly at the early stages of virus infection. These findings highlight Wolbachia's ability to alter its gene expression in response to DENV infection of the host cell. IMPORTANCE Aedes aegypti is a vector of several pathogenic viruses, including dengue, Zika, chikungunya, and yellow fever viruses, which are of importance to human health. Wolbachia is an endosymbiotic bacterium currently used in transinfected mosquitoes to suppress replication and transmission of dengue viruses. However, the mechanism of Wolbachia-mediated virus inhibition is not fully understood. While several studies have shown mosquitoes' transcriptional responses to dengue virus infection, none have investigated these responses in Wolbachia, which may provide clues to the inhibition mechanism. Our results suggest changes in the expression of a number of functionally important Wolbachia genes upon dengue virus infection, including those involved in stress responses, providing insights into the endosymbiont's reaction to virus infection.}, } @article {pmid34188180, year = {2021}, author = {Zhang, Y and Cai, T and Ren, Z and Liu, Y and Yuan, M and Cai, Y and Yu, C and Shu, R and He, S and Li, J and Wong, ACN and Wan, H}, title = {Decline in symbiont-dependent host detoxification metabolism contributes to increased insecticide susceptibility of insects under high temperature.}, journal = {The ISME journal}, volume = {15}, number = {12}, pages = {3693-3703}, pmid = {34188180}, issn = {1751-7370}, mesh = {Animals ; Drosophila melanogaster ; *Hemiptera ; Insecta ; Insecticide Resistance ; *Insecticides/toxicity ; Temperature ; }, abstract = {The interactions between insects and their bacterial symbionts are shaped by a variety of abiotic factors, including temperature. As global temperatures continue to break high records, a great deal of uncertainty surrounds how agriculturally important insect pests and their symbionts may be affected by elevated temperatures, and its implications for future pest management. In this study, we examine the role of bacterial symbionts in the brown planthopper Nilaparvata lugens response to insecticide (imidacloprid) under different temperature scenarios. Our results reveal that the bacterial symbionts orchestrate host detoxification metabolism via the CncC pathway to promote host insecticide resistance, whereby the symbiont-inducible CncC pathway acts as a signaling conduit between exogenous abiotic stimuli and host metabolism. However, this insect-bacterial partnership function is vulnerable to high temperature, which causes a significant decline in host-bacterial content. In particular, we have identified the temperature-sensitive Wolbachia as a candidate player in N. lugens detoxification metabolism. Wolbachia-dependent insecticide resistance was confirmed through a series of insecticide assays and experiments comparing Wolbachia-free and Wolbachia-infected N. lugens and also Drosophila melanogaster. Together, our research reveals elevated temperatures negatively impact insect-bacterial symbiosis, triggering adverse consequences on host response to insecticide (imidacloprid) and potentially other xenobiotics.}, } @article {pmid34156702, year = {2021}, author = {Cruz, MA and Magalhães, S and Sucena, É and Zélé, F}, title = {Wolbachia and host intrinsic reproductive barriers contribute additively to postmating isolation in spider mites.}, journal = {Evolution; international journal of organic evolution}, volume = {75}, number = {8}, pages = {2085-2101}, doi = {10.1111/evo.14286}, pmid = {34156702}, issn = {1558-5646}, mesh = {Animals ; Diploidy ; *Mites ; Reproduction ; Reproductive Isolation ; Symbiosis ; *Tetranychidae/genetics ; *Wolbachia ; }, abstract = {Wolbachia are maternally-inherited bacteria that induce cytoplasmic incompatibility in many arthropod species. However, the ubiquity of this isolation mechanism for host speciation processes remains elusive, as only few studies have examined Wolbachia-induced incompatibilities when host populations are not genetically compatible. Here, we used three populations of two genetically differentiated colour forms of the haplodiploid spider mite Tetranychus urticae to dissect the interaction between Wolbachia-induced and host-associated incompatibilities, and their relative contribution to postmating isolation. We found that these two sources of incompatibility act through different mechanisms in an additive fashion. Host-associated incompatibility contributes 1.5 times more than Wolbachia-induced incompatibility in reducing hybrid production, the former through an overproduction of haploid sons at the expense of diploid daughters (ca. 75% decrease) and the latter by increasing the embryonic mortality of daughters (by ca. 49%). Furthermore, regardless of cross direction, we observed near-complete F1 hybrid sterility and complete F2 hybrid breakdown between populations of the two forms, but Wolbachia did not contribute to this outcome. We thus show mechanistic independence and an additive nature of host-intrinsic and Wolbachia-induced sources of isolation. Wolbachia may contribute to reproductive isolation in this system, thereby potentially affecting host differentiation and distribution in the field.}, } @article {pmid34152527, year = {2021}, author = {Dângelo, RAC and Michereff-Filho, M and Inoue-Nagata, AK and da Silva, PS and Chediak, M and Guedes, RNC}, title = {Area-wide insecticide resistance and endosymbiont incidence in the whitefly Bemisia tabaci MEAM1 (B biotype): A Neotropical context.}, journal = {Ecotoxicology (London, England)}, volume = {30}, number = {6}, pages = {1056-1070}, pmid = {34152527}, issn = {1573-3017}, mesh = {Animals ; *Hemiptera ; Humans ; Incidence ; Insecticide Resistance ; *Insecticides/toxicity ; Symbiosis ; }, abstract = {Agriculture insecticides are used against insect pest species, but are able to change community structure in contaminated habitats, and also the genetic pool of exposed individuals. In fact, the latter effect is a relevant tool to in situ biomonitoring of pollutant contamination and impact, besides its practical economic and management concerns. This takes place because the emergence of individuals with resistance to insecticides is particularly frequent among insect pest species and usually enhances insecticide overuse and crop losses. Pest insects of global prominence such as whiteflies are a focus of attention due to problems with insecticide resistance and association with endosymbionts, as the case of the invasive putative species Bemisia tabaci MEAM1. The scenario is particularly complex in the Neotropics, where insecticide use is ubiquitous, but whose spatial scale of occurrence is usually neglected. Here we explored the spatial-dependence of both phenomena in MEAM1 whiteflies recording resistance to two widely used insecticides, lambda-cyhalothrin and spiromesifen, and endosymbiont co-occurrence. Resistance to both insecticides was frequent exhibiting low to moderate frequency of lambda-cyhalothrin resistance and moderate to high frequency of spiromesifen resistance. Among the prevailing whitefly endosymbionts, Wolbachia, Cardinium and Arsenophonus were markedly absent. In contrast, Hamiltonella and Rickettsia prevailed and their incidence was correlated. Furthermore, Rickettsia endosymbionts were particularly associated with lambda-cyhalothrin susceptibility. These traits were spatially dependent with significant variation taking place within an area of about 700 Km[2]. Such findings reinforce the notion of endosymbiont-associated resistance to insecticides, and also of their local incidence allowing spatial mapping and locally-targeted mitigation.}, } @article {pmid34149963, year = {2021}, author = {Ponce, GE and Fuse, M and Chan, A and Connor, EF}, title = {The Localization of Phytohormones within the Gall-inducing Insect Eurosta solidaginis (Diptera: Tephritidae).}, journal = {Arthropod-plant interactions}, volume = {15}, number = {3}, pages = {375-385}, pmid = {34149963}, issn = {1872-8855}, support = {R25 GM048972/GM/NIGMS NIH HHS/United States ; }, abstract = {The phytohormone production hypothesis suggests that organisms, including insects, induce galls by producing and secreting plant growth hormones. Auxins and cytokinins are classes of phytohormones that induce cell growth and cell division, which could contribute to the plant tissue proliferation which constitutes the covering gall. Bacteria, symbiotic with insects, may also play a part in gall induction by insects through the synthesis of phytohormones or other effectors. Past studies have shown that concentrations of cytokinins and auxins in gall-inducing insects are higher than in their host plants. However, these analyses have involved whole-body extractions. Using immunolocalization of cytokinin and auxin, in the gall inducing stage of Eurosta solidaginis, we found both phytohormones to localize almost exclusively to the salivary glands. Co-localization of phytohormone label with a nucleic acid stain in the salivary glands revealed the absence of Wolbachia sp., the bacterial symbiont of E. solidaginis, which suggests that phytohormone production is symbiont independent. Our findings are consistent with the hypothesis that phytohormones are synthesized in and secreted from the salivary glands of E. solidaginis into host-plant tissues for the purpose of manipulating the host plant.}, } @article {pmid34145422, year = {2021}, author = {York, A}, title = {Defeating dengue with Wolbachia.}, journal = {Nature reviews. Microbiology}, volume = {19}, number = {8}, pages = {482}, pmid = {34145422}, issn = {1740-1534}, } @article {pmid34143770, year = {2021}, author = {Duarte, EH and Carvalho, A and López-Madrigal, S and Costa, J and Teixeira, L}, title = {Forward genetics in Wolbachia: Regulation of Wolbachia proliferation by the amplification and deletion of an addictive genomic island.}, journal = {PLoS genetics}, volume = {17}, number = {6}, pages = {e1009612}, pmid = {34143770}, issn = {1553-7404}, mesh = {Animals ; Bacterial Load ; Dicistroviridae/growth & development/pathogenicity ; Drosophila melanogaster/immunology/*microbiology/virology ; Female ; Gene Editing/methods ; *Genome, Bacterial ; Genomic Islands ; Longevity/*immunology ; Male ; Phenotype ; Symbiosis/*genetics ; Wolbachia/*genetics/growth & development/metabolism ; }, abstract = {Wolbachia is one of the most prevalent bacterial endosymbionts, infecting approximately 40% of terrestrial arthropod species. Wolbachia is often a reproductive parasite but can also provide fitness benefits to its host, as, for example, protection against viral pathogens. This protective effect is currently being applied to fight arboviruses transmission by releasing Wolbachia-transinfected mosquitoes. Titre regulation is a crucial aspect of Wolbachia biology. Higher titres can lead to stronger phenotypes and fidelity of transmission but can have a higher cost to the host. Since Wolbachia is maternally transmitted, its fitness depends on host fitness, and, therefore, its cost to the host may be under selection. Understanding how Wolbachia titres are regulated and other aspects of Wolbachia biology has been hampered by the lack of genetic tools. Here we developed a forward genetic screen to identify new Wolbachia over-proliferative mutant variants. We characterized in detail two new mutants, wMelPop2 and wMelOctoless, and show that the amplification or loss of the Octomom genomic region lead to over-proliferation. These results confirm previous data and expand on the complex role of this genomic region in the control of Wolbachia proliferation. Both new mutants shorten the host lifespan and increase antiviral protection. Moreover, we show that Wolbachia proliferation rate in Drosophila melanogaster depends on the interaction between Octomom copy number, the host developmental stage, and temperature. Our analysis also suggests that the life shortening and antiviral protection phenotypes of Wolbachia are dependent on different, but related, properties of the endosymbiont; the rate of proliferation and the titres near the time of infection, respectively. We also demonstrate the feasibility of a novel and unbiased experimental approach to study Wolbachia biology, which could be further adapted to characterize other genetically intractable bacterial endosymbionts.}, } @article {pmid34125999, year = {2022}, author = {Norte, AC and Harris, DJ and Silveira, D and Nunes, CS and Núncio, MS and Martínez, EG and Giménez, A and de Sousa, R and Lopes de Carvalho, I and Perera, A}, title = {Diversity of microorganisms in Hyalomma aegyptium collected from spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia.}, journal = {Transboundary and emerging diseases}, volume = {69}, number = {4}, pages = {1951-1962}, doi = {10.1111/tbed.14188}, pmid = {34125999}, issn = {1865-1682}, mesh = {Animals ; *Borrelia ; Ehrlichia ; Humans ; *Ixodidae/microbiology ; *Rickettsia/genetics ; *Ticks/microbiology ; Tunisia/epidemiology ; Turkey/epidemiology ; *Turtles/parasitology ; }, abstract = {Ticks carry a diverse community of microorganisms including non-pathogenic symbionts, commensals, and pathogens, such as viruses, bacteria, protozoans, and fungi. The assessment of tick-borne microorganisms (TBM) in tortoises and their ticks is essential to understand their eco-epidemiology, and to map and monitor potential pathogens to humans and other animals. The aim of this study was to characterize the diversity of microorganisms found in ticks collected from the spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia. Ticks feeding on wild T. graeca were collected, and pathogens were screened by polymerase chain reaction using group-specific primers. In total, 131 adult Hyalomma aegyptium ticks were collected from 92 T. graeca in Morocco (n = 48), Tunisia (n = 2), Algeria (n = 70), and Turkey (n = 11). Bacteria and protozoa detected included Hemolivia mauritanica (22.9%), Midichloria mitochondrii (11.4%), relapsing-fever borreliae (8.4%), Ehrlichia spp. (7.6%), Rickettsia spp. (3.4%), Borrelia burgdorferi s.l. (0.9%), Francisella spp. (0.9%), and Wolbachia spp. (0.8%). The characterization of Rickettsia included R. sibirica mongolitimonae (Algeria), R. aeschlimannii (Turkey), and R.africae (Morocco). Hemolivia mauritanica and Ehrlichia spp. prevalence varied significantly with the sampling region/country. We did not detect significant associations in microorganism presence within ticks, nor between microorganism presence and tick mitochondrial DNA haplogroups. This is the first report of Francisella persica-like, relapsing fever borreliae, M. mitochondrii, and Wolbachia spp. in H. aegyptium ticks collected from wild hosts from the South and Eastern Mediterranean region, and of R. sibirica mongolitimonae and R. africae in H. aegyptium from Algeria and Morocco, respectively. Given that T. graeca is a common species in commercial and non-commercial pet trade, the evaluation of the role of this species and its ticks as hosts for TBM is particularly relevant for public health.}, } @article {pmid34122367, year = {2021}, author = {Demirbas-Uzel, G and Augustinos, AA and Doudoumis, V and Parker, AG and Tsiamis, G and Bourtzis, K and Abd-Alla, AMM}, title = {Interactions Between Tsetse Endosymbionts and Glossina pallidipes Salivary Gland Hypertrophy Virus in Glossina Hosts.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {653880}, pmid = {34122367}, issn = {1664-302X}, abstract = {Tsetse flies are the sole cyclic vector for trypanosomosis, the causative agent for human African trypanosomosis or sleeping sickness and African animal trypanosomosis or nagana. Tsetse population control is the most efficient strategy for animal trypanosomosis control. Among all tsetse control methods, the Sterile Insect Technique (SIT) is one of the most powerful control tactics to suppress or eradicate tsetse flies. However, one of the challenges for the implementation of SIT is the mass production of target species. Tsetse flies have a highly regulated and defined microbial fauna composed of three bacterial symbionts (Wigglesworthia, Sodalis and Wolbachia) and a pathogenic Glossina pallidipes Salivary Gland Hypertrophy Virus (GpSGHV) which causes reproduction alterations such as testicular degeneration and ovarian abnormalities with reduced fertility and fecundity. Interactions between symbionts and GpSGHV might affect the performance of the insect host. In the present study, we assessed the possible impact of GpSGHV on the prevalence of tsetse endosymbionts under laboratory conditions to decipher the bidirectional interactions on six Glossina laboratory species. The results indicate that tsetse symbiont densities increased over time in tsetse colonies with no clear impact of the GpSGHV infection on symbionts density. However, a positive correlation between the GpSGHV and Sodalis density was observed in Glossina fuscipes species. In contrast, a negative correlation between the GpSGHV density and symbionts density was observed in the other taxa. It is worth noting that the lowest Wigglesworthia density was observed in G. pallidipes, the species which suffers most from GpSGHV infection. In conclusion, the interactions between GpSGHV infection and tsetse symbiont infections seems complicated and affected by the host and the infection density of the GpSGHV and tsetse symbionts.}, } @article {pmid34110943, year = {2021}, author = {Christensen, D and Khoshmanesh, A and Perez-Guaita, D and Iturbe-Ormaetxe, I and O'Neill, S and Wood, BR}, title = {Detection and Identification of Wolbachia pipientis Strains in Mosquito Eggs Using Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy.}, journal = {Applied spectroscopy}, volume = {75}, number = {8}, pages = {1003-1011}, doi = {10.1177/00037028211027140}, pmid = {34110943}, issn = {1943-3530}, mesh = {*Aedes ; Animals ; Fourier Analysis ; Mosquito Vectors ; Spectroscopy, Fourier Transform Infrared ; *Wolbachia ; }, abstract = {The global fight against mosquito-borne viral diseases has in recent years been bolstered by the introduction of the endosymbiotic bacteria Wolbachia to vector populations, which in host mosquitoes suppresses the transmissibility of several viruses. Researchers engaged on this front of the battle often need to know the Wolbachia infection status of individual mosquitoes, as the intervention progresses and the mosquitoes become established in the target population. Previously, we successfully applied attenuated total reflection Fourier transform infrared spectroscopy to the detection of Wolbachia in adult Aedes aegypti mosquitoes; here we apply the same principles to Aedes eggs, with sensitivity and selectivity > 0.95. Further, we successfully distinguish between infections in eggs of the wMel and wMelPop strains of Wolbachia pipientis, with a classification error of 3%. The disruption of host lipid profile by Wolbachia is found to be a key driver in spectral differences between these sample classes.}, } @article {pmid34108021, year = {2021}, author = {Novelo, M and Audsley, MD and McGraw, EA}, title = {The effects of DENV serotype competition and co-infection on viral kinetics in Wolbachia-infected and uninfected Aedes aegypti mosquitoes.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {314}, pmid = {34108021}, issn = {1756-3305}, support = {APP1103804//National Health and Medical Research Council/ ; AI143758//Foundation for the National Institutes of Health/ ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Dengue Virus/chemistry/classification/genetics/*physiology ; Female ; Kinetics ; Mosquito Vectors/*microbiology/physiology/*virology ; Viral Load ; Virus Replication ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: The Aedes aegypti mosquito is responsible for the transmission of several medically important arthropod-borne viruses, including multiple serotypes of dengue virus (DENV-1, -2, -3, and -4). Competition within the mosquito between DENV serotypes can affect viral infection dynamics, modulating the transmission potential of the pathogen. Vector control remains the main method for limiting dengue fever. The insect endosymbiont Wolbachia pipientis is currently being trialed in field releases globally as a means of biological control because it reduces virus replication inside the mosquito. It is not clear how co-infection between DENV serotypes in the same mosquito might alter the pathogen-blocking phenotype elicited by Wolbachia in Ae. aegypti.

METHODS: Five- to 7-day-old female Ae. aegypti from two lines, namely, with (wMel) and without Wolbachia infection (WT), were fed virus-laden blood through an artificial membrane with either a mix of DENV-2 and DENV-3 or the same DENV serotypes singly. Mosquitoes were subsequently incubated inside environmental chambers and collected on the following days post-infection: 3, 4, 5, 7, 8, 9, 11, 12, and 13. Midgut, carcass, and salivary glands were collected from each mosquito at each timepoint and individually analyzed to determine the percentage of DENV infection and viral RNA load via RT-qPCR.

RESULTS: We saw that for WT mosquitoes DENV-3 grew to higher viral RNA loads across multiple tissues when co-infected with DENV-2 than when it was in a mono-infection. Additionally, we saw a strong pathogen-blocking phenotype in wMel mosquitoes independent of co-infection status.

CONCLUSION: In this study, we demonstrated that the wMel mosquito line is capable of blocking DENV serotype co-infection in a systemic way across the mosquito body. Moreover, we showed that for WT mosquitoes, serotype co-infection can affect infection frequency in a tissue- and time-specific manner and that both viruses have the potential of being transmitted simultaneously. Our findings suggest that the long-term efficacy of Wolbachia pathogen blocking is not compromised by arthropod-borne virus co-infection.}, } @article {pmid34107180, year = {2021}, author = {Utarini, A and Indriani, C and Ahmad, RA and Tantowijoyo, W and Arguni, E and Ansari, MR and Supriyati, E and Wardana, DS and Meitika, Y and Ernesia, I and Nurhayati, I and Prabowo, E and Andari, B and Green, BR and Hodgson, L and Cutcher, Z and Rancès, E and Ryan, PA and O'Neill, SL and Dufault, SM and Tanamas, SK and Jewell, NP and Anders, KL and Simmons, CP and , }, title = {Efficacy of Wolbachia-Infected Mosquito Deployments for the Control of Dengue.}, journal = {The New England journal of medicine}, volume = {384}, number = {23}, pages = {2177-2186}, pmid = {34107180}, issn = {1533-4406}, support = {/WT_/Wellcome Trust/United Kingdom ; 212914/WT_/Wellcome Trust/United Kingdom ; R01 AI148127/AI/NIAID NIH HHS/United States ; }, mesh = {Adolescent ; Adult ; Aedes/*microbiology/virology ; Animals ; Child ; Child, Preschool ; Communicable Disease Control/*methods ; Dengue/diagnosis/epidemiology/prevention & control/*transmission ; Dengue Virus/isolation & purification ; Female ; Humans ; Incidence ; Indonesia/epidemiology ; Male ; Middle Aged ; *Mosquito Vectors/microbiology/virology ; *Wolbachia ; Young Adult ; }, abstract = {BACKGROUND: Aedes aegypti mosquitoes infected with the wMel strain of Wolbachia pipientis are less susceptible than wild-type A. aegypti to dengue virus infection.

METHODS: We conducted a cluster-randomized trial involving releases of wMel-infected A. aegypti mosquitoes for the control of dengue in Yogyakarta, Indonesia. We randomly assigned 12 geographic clusters to receive deployments of wMel-infected A. aegypti (intervention clusters) and 12 clusters to receive no deployments (control clusters). All clusters practiced local mosquito-control measures as usual. A test-negative design was used to assess the efficacy of the intervention. Patients with acute undifferentiated fever who presented to local primary care clinics and were 3 to 45 years of age were recruited. Laboratory testing was used to identify participants who had virologically confirmed dengue (VCD) and those who were test-negative controls. The primary end point was symptomatic VCD of any severity caused by any dengue virus serotype.

RESULTS: After successful introgression of wMel into the intervention clusters, 8144 participants were enrolled; 3721 lived in intervention clusters, and 4423 lived in control clusters. In the intention-to-treat analysis, VCD occurred in 67 of 2905 participants (2.3%) in the intervention clusters and in 318 of 3401 (9.4%) in the control clusters (aggregate odds ratio for VCD, 0.23; 95% confidence interval [CI], 0.15 to 0.35; P = 0.004). The protective efficacy of the intervention was 77.1% (95% CI, 65.3 to 84.9) and was similar against the four dengue virus serotypes. The incidence of hospitalization for VCD was lower among participants who lived in intervention clusters (13 of 2905 participants [0.4%]) than among those who lived in control clusters (102 of 3401 [3.0%]) (protective efficacy, 86.2%; 95% CI, 66.2 to 94.3).

CONCLUSIONS: Introgression of wMel into A. aegypti populations was effective in reducing the incidence of symptomatic dengue and resulted in fewer hospitalizations for dengue among the participants. (Funded by the Tahija Foundation and others; AWED ClinicalTrials.gov number, NCT03055585; Indonesia Registry number, INA-A7OB6TW.).}, } @article {pmid34102127, year = {2021}, author = {Ross, PA and Hoffmann, AA}, title = {Vector control: Discovery of Wolbachia in malaria vectors.}, journal = {Current biology : CB}, volume = {31}, number = {11}, pages = {R738-R740}, doi = {10.1016/j.cub.2021.04.038}, pmid = {34102127}, issn = {1879-0445}, mesh = {*Aedes ; Animals ; *Anopheles ; *Malaria/prevention & control ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Wolbachia bacteria are being widely released for suppression of dengue transmitted by Aedes mosquitoes. Walker, Quek, Jeffries and colleagues present robust evidence for natural Wolbachia infections in malaria-vectoring Anopheles mosquitoes, paving the way for new Wolbachia-based interventions.}, } @article {pmid34091278, year = {2021}, author = {Dall'Agnol, B and McCulloch, JA and Mayer, FQ and Souza, U and Webster, A and Antunes, P and Doyle, RL and Reck, J and Ferreira, CAS}, title = {Molecular characterization of bacterial communities of two neotropical tick species (Amblyomma aureolatum and Ornithodoros brasiliensis) using rDNA 16S sequencing.}, journal = {Ticks and tick-borne diseases}, volume = {12}, number = {5}, pages = {101746}, doi = {10.1016/j.ttbdis.2021.101746}, pmid = {34091278}, issn = {1877-9603}, mesh = {Amblyomma/*microbiology ; Animals ; Bacteria/genetics/isolation & purification ; Coxiella/genetics/isolation & purification ; DNA, Bacterial/isolation & purification ; Francisella/genetics/isolation & purification ; Ixodidae/microbiology ; Metagenomics ; *Microbiota ; Ornithodoros/*microbiology ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/isolation & purification ; }, abstract = {Ticks are one of the main vectors of pathogens for humans and animals worldwide. However, they harbor non-pathogenic microorganisms that are important for their survival, facilitating both their nutrition and immunity. We investigated the bacterial communities associated with two neotropical tick species of human and veterinary potential health importance from Brazil: Amblyomma aureolatum and Ornithodoros brasiliensis. In A. aureolatum (adult ticks collected from wild canids from Southern Brazil), the predominant bacterial phyla were Proteobacteria (98.68%), Tenericutes (0.70%), Bacteroidetes (0.14%), Actinobacteria (0.13%), and Acidobacteria (0.05%). The predominant genera were Francisella (97.01%), Spiroplasma (0.70%), Wolbachia (0.51%), Candidatus Midichloria (0.25%), and Alkanindiges (0.13%). The predominant phyla in O. brasiliensis (adults, fed and unfed nymphs collected at the environment from Southern Brazil) were Proteobacteria (90.27%), Actinobacteria (7.38%), Firmicutes (0.77%), Bacteroidetes (0.44%), and Planctomycetes (0.22%). The predominant bacterial genera were Coxiella (87.71%), Nocardioides (1.73%), Saccharopolyspora (0.54%), Marmoricola (0.42%), and Staphylococcus (0.40%). Considering the genera with potential importance for human and animal health which can be transmitted by ticks, Coxiella sp. was found in all stages of O. brasiliensis, Francisella sp. in all stages of A. aureolatum and in unfed nymphs of O. brasiliensis, and Rickettsia sp. in females of A. aureolatum from Banhado dos Pachecos (BP) in Viamão municipality, Brazil, and in females and unfed nymphs of O. brasiliensis. These results deepen our understanding of the tick-microbiota relationship in Ixodidae and Argasidae, driving new studies with the focus on the manipulation of tick microbiota to prevent outbreaks of tick-borne diseases in South America.}, } @article {pmid34086908, year = {2021}, author = {Farahani, HK and Ashouri, A and Abroon, P and Pierre, JS and van Baaren, J}, title = {Wolbachia manipulate fitness benefits of olfactory associative learning in a parasitoid wasp.}, journal = {The Journal of experimental biology}, volume = {224}, number = {11}, pages = {}, doi = {10.1242/jeb.240549}, pmid = {34086908}, issn = {1477-9145}, mesh = {Animals ; Conditioning, Classical ; Female ; Memory ; Smell ; *Wasps ; *Wolbachia ; }, abstract = {Upon encountering a host, a female parasitoid wasp has to decide whether to learn positive or negative cues related to the host. The optimal female decision will depend on the fitness costs and benefits of learned stimuli. Reward quality is positively related to the rate of behavioral acquisition in processes such as associative learning. Wolbachia, an endosymbiotic bacterium, often plays an impressive role in the manipulation of its arthropod host's biology. Here, we studied the responses of two natural Wolbachia infected/uninfected Trichogramma brassicae wasp populations to theoretically high- and low-reward values during a conditioning process and the consequences of their responses in terms of memory duration. According to our results, uninfected wasps showed an attraction response to high-value rewards, but showed aversive learning in response to low-value rewards. The memory span of uninfected wasps after conditioning by low-value rewards was significantly shorter than that for high-value rewards. As our results revealed, responses to high-quality hosts will bring more benefits (bigger size, increased fecundity and enhanced survival) than those to low-quality hosts for uninfected wasps. Infected wasps were attracted to conditioned stimuli with the same memory duration after conditioning by both types of hosts. This was linked to the fact that parasitoids emerging from both types of hosts present the same life-history traits. Therefore, these hosts represent the same quality reward for infected wasps. According to the obtained results, it can be concluded that Wolbachia manipulates the learning ability of its host, resulting in the wasp responding to all reward values similarly.}, } @article {pmid34079424, year = {2021}, author = {Ta-Tang, TH and Luz, SLB and Crainey, JL and Rubio, JM}, title = {An Overview of the Management of Mansonellosis.}, journal = {Research and reports in tropical medicine}, volume = {12}, number = {}, pages = {93-105}, pmid = {34079424}, issn = {1179-7282}, abstract = {Mansonellosis is caused by three filarial parasite species from the genus Mansonella that commonly produce chronic human microfilaraemias: M. ozzardi, M. perstans and M. streptocerca. The disease is widespread in Africa, the Caribbean and South and Central America, and although it is typically asymptomatic it has been associated with mild pathologies including leg-chills, joint-pains, headaches, fevers, and corneal lesions. No robust mansonellosis disease burden estimates have yet been made and the impact the disease has on blood bank stocks and the monitoring of other filarial diseases is not thought to be of sufficient public health importance to justify dedicated disease management interventions. Mansonellosis´s Ceratopogonidae and Simuliidae vectors are not targeted by other control programmes and because of their small size and out-door biting habits are unlikely to be affected by interventions targeting other disease vectors like mosquitoes. The ivermectin and mebendazole-based mass drug administration (iMDA and mMDA) treatment regimens deployed by the WHO´s Elimination of Neglected Tropical Diseases (ESPEN) programme and its forerunners have, however, likely impacted significantly on the mansonellosis disease burden, principally by reducing the transmission of M. streptocerca in Africa. The increasingly popular plan of using iMDA to control malaria could also affect M. ozzardi parasite prevalence and transmission in Latin America in the future. However, a potentially far greater mansonellosis disease burden impact is likely to come from short-course curative anti-Wolbachia therapeutics, which are presently being developed for onchocerciasis and lymphatic filariasis treatment. Even if the WHO´s ESPEN programme does not choose to deploy these drugs in MDA interventions, they have the potential to dramatically increase the financial and logistical feasibility of effective mansonellosis management. There is, thus, now a fresh and urgent need to better characterise the disease burden and eco-epidemiology of mansonellosis so that effective management programmes can be designed, advocated for and implemented.}, } @article {pmid34074789, year = {2021}, author = {Baker, BM and Carbone, MA and Huang, W and Anholt, RRH and Mackay, TFC}, title = {Genetic basis of variation in cocaine and methamphetamine consumption in outbred populations of Drosophila melanogaster.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {118}, number = {23}, pages = {}, pmid = {34074789}, issn = {1091-6490}, support = {U01 DA041613/DA/NIDA NIH HHS/United States ; }, mesh = {Animals ; Cocaine/*pharmacology ; Drosophila Proteins/*genetics/metabolism ; Drosophila melanogaster ; *Epistasis, Genetic ; Female ; Genome-Wide Association Study ; Humans ; Male ; Methamphetamine/*pharmacology ; *Polymorphism, Single Nucleotide ; *Quantitative Trait Loci ; *Sex Characteristics ; }, abstract = {We used Drosophila melanogaster to map the genetic basis of naturally occurring variation in voluntary consumption of cocaine and methamphetamine. We derived an outbred advanced intercross population (AIP) from 37 sequenced inbred wild-derived lines of the Drosophila melanogaster Genetic Reference Panel (DGRP), which are maximally genetically divergent, have minimal residual heterozygosity, are not segregating for common inversions, and are not infected with Wolbachia pipientis We assessed consumption of sucrose, methamphetamine-supplemented sucrose, and cocaine-supplemented sucrose and found considerable phenotypic variation for consumption of both drugs, in both sexes. We performed whole-genome sequencing and extreme quantitative trait locus (QTL) mapping on the top 10% of consumers for each replicate, sex, and condition and an equal number of randomly selected flies. We evaluated changes in allele frequencies among high consumers and control flies and identified 3,033 variants significantly (P < 1.9 × 10[-8]) associated with increased consumption, located in or near 1,962 genes. Many of these genes are associated with nervous system development and function, and 77 belong to a known gene-gene interaction subnetwork. We assessed the effects of RNA interference (RNAi) on drug consumption for 22 candidate genes; 17 had a significant effect in at least one sex. We constructed allele-specific AIPs that were homozygous for alternative candidate alleles for 10 single-nucleotide polymorphisms (SNPs) and measured average consumption for each population; 9 SNPs had significant effects in at least one sex. The genetic basis of voluntary drug consumption in Drosophila is polygenic and implicates genes with human orthologs and associated variants with sex- and drug-specific effects.}, } @article {pmid34074606, year = {2021}, author = {Schmidt, TL and Endersby-Harshman, NM and Hoffmann, AA}, title = {Improving mosquito control strategies with population genomics.}, journal = {Trends in parasitology}, volume = {37}, number = {10}, pages = {907-921}, doi = {10.1016/j.pt.2021.05.002}, pmid = {34074606}, issn = {1471-5007}, mesh = {Animals ; *Culicidae/genetics ; Genome, Insect/genetics ; *Genomics ; Insecticide Resistance/genetics ; *Mosquito Control/methods ; }, abstract = {Mosquito control strategies increasingly apply knowledge from population genomics research. This review highlights recent applications to three research domains: mosquito invasions, insecticide resistance evolution, and rear and release programs. Current research trends follow developments in reference assemblies, either as improvements to existing assemblies (particularly Aedes) or assemblies for new taxa (particularly Anopheles). With improved assemblies, studies of invasive and rear and release target populations are better able to incorporate adaptive as well as demographic hypotheses. New reference assemblies are aiding comparisons of insecticide resistance across sister taxa while helping resolve taxon boundaries amidst frequent introgression. Anopheles gene drive deployments and improved Aedes genome assemblies should lead to a convergence in research aims for Anopheles and Aedes in the coming years.}, } @article {pmid34066350, year = {2021}, author = {Liu, Q and Zhang, H and Zeng, L and Yu, Y and Lin, X and Huang, X}, title = {Coexistence of Three Dominant Bacterial Symbionts in a Social Aphid and Implications for Ecological Adaptation.}, journal = {Insects}, volume = {12}, number = {5}, pages = {}, pmid = {34066350}, issn = {2075-4450}, support = {31970446//National Natural Science Foundation of China/ ; 2016YFE0203100//National Key R&D Program of China/ ; }, abstract = {Aphids are associated with an array of symbionts that have diverse ecological and evolutionary effects on their hosts. To date, symbiont communities of most aphid species are still poorly characterized, especially for the social aphids. In this study, high-throughput 16S rDNA amplicon sequencing was used to assess the bacterial communities of the social aphid Pseudoregma bambucicola, and the differences in bacterial diversity with respect to ant attendance and time series were also assessed. We found that the diversity of symbionts in P. bambucicola was low and three dominant symbionts (Buchnera, Pectobacterium and Wolbachia) were stably coexisting. Pectobacterium may help P. bambucicola feed on the hard bamboo stems, and genetic distance analysis suggests that the Pectobacterium in P. bambucicola may be a new symbiont species. Wolbachia may be associated with the transition of reproduction mode or has a nutritional role in P. bambucicola. Statistical tests on the diversity of bacterial communities in P. bambucicola suggest that aphid populations attended by ants usually have a significantly higher evenness than populations without ant attendance but there was no significant difference among aphid populations from different seasons.}, } @article {pmid34059765, year = {2021}, author = {Detcharoen, M and Schilling, MP and Arthofer, W and Schlick-Steiner, BC and Steiner, FM}, title = {Differential gene expression in Drosophila melanogaster and D. nigrosparsa infected with the same Wolbachia strain.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {11336}, pmid = {34059765}, issn = {2045-2322}, mesh = {Animals ; Drosophila melanogaster/*metabolism/*microbiology ; Female ; *Gene Expression ; Gene Expression Profiling ; Species Specificity ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited endosymbionts that infect nearly half of all arthropod species. Wolbachia manipulate their hosts to maximize their transmission, but they can also provide benefits such as nutrients and resistance against viruses to their hosts. The Wolbachia strain wMel was recently found to increase locomotor activities and possibly trigger cytoplasmic incompatibility in the transinfected fly Drosophila nigrosparsa. Here, we investigated, in females of both D. melanogaster and D. nigrosparsa, the gene expression between animals uninfected and infected with wMel, using RNA sequencing to see if the two Drosophila species respond to the infection in the same or different ways. A total of 2164 orthologous genes were used. The two fly species responded to the infection in different ways. Significant changes shared by the fly species belong to the expression of genes involved in processes such as oxidation-reduction process, iron-ion binding, and voltage-gated potassium-channel activity. We discuss our findings also in the light of how Wolbachia survive within both the native and the novel host.}, } @article {pmid34047357, year = {2021}, author = {Ün, Ç and Schultner, E and Manzano-Marín, A and Flórez, LV and Seifert, B and Heinze, J and Oettler, J}, title = {Cytoplasmic incompatibility between Old and New World populations of a tramp ant.}, journal = {Evolution; international journal of organic evolution}, volume = {75}, number = {7}, pages = {1775-1791}, doi = {10.1111/evo.14261}, pmid = {34047357}, issn = {1558-5646}, mesh = {Animals ; *Ants/genetics ; Biological Evolution ; Cytoplasm ; Reproduction ; Symbiosis ; *Wolbachia ; }, abstract = {Reproductive manipulation by endosymbiotic Wolbachia can cause unequal inheritance, allowing the manipulator to spread and potentially impacting evolutionary dynamics in infected hosts. Tramp and invasive species are excellent models to study the dynamics of host-Wolbachia associations because introduced populations often diverge in their microbiomes after colonizing new habitats, resulting in infection polymorphisms between native and introduced populations. Ants are the most abundant group of insects on earth, and numerous ant species are classified as highly invasive. However, little is known about the role of Wolbachia in these ecologically dominant insects. Here, we provide the first description of reproductive manipulation by Wolbachia in an ant. We show that Old and New World populations of the cosmotropic tramp ant Cardiocondyla obscurior harbor distinct Wolbachia strains, and that only the Old World strain manipulates host reproduction by causing cytoplasmic incompatibility (CI) in hybrid crosses. By uncovering a symbiont-induced mechanism of reproductive isolation in a social insect, our study provides a novel perspective on the biology of tramp ants and introduces a new system for studying the evolutionary consequences of CI.}, } @article {pmid34034166, year = {2021}, author = {Li, S and Li, J and Li, Z and Ke, X and Wu, L and Christie, P}, title = {Toxic effects of norfloxacin in soil on fed and unfed Folsomia candida (Isotomidae: Collembola) and on gut and soil microbiota.}, journal = {The Science of the total environment}, volume = {788}, number = {}, pages = {147793}, doi = {10.1016/j.scitotenv.2021.147793}, pmid = {34034166}, issn = {1879-1026}, mesh = {Animals ; *Arthropods ; *Microbiota ; Norfloxacin/toxicity ; Soil ; *Soil Pollutants/toxicity ; }, abstract = {Soils contaminated with antibiotics may exert effects on soil-dwelling animals. A systematic ecological toxicity assessment of norfloxacin on the soil collembolan Folsomia candida (F. candida) was therefore conducted in soil and Petri dish systems with and without feeding at the population, individual and cellular levels. The indicators survival, reproduction, antioxidant enzyme activities peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD), malondialdehyde (MDA) contents and gut microbiota were studied. The surrounding soil microbiota were also investigated because F. candida can ingest soil microbiota that may have effects on the gut microbiota. In general, the toxicity of norfloxacin to F. candida in contaminated soil without food addition was higher than in contaminated soil with food addition. Norfloxacin had little effect at population and individual levels but antioxidant enzyme activities changed significantly in treatments with longer exposure times or higher norfloxacin concentrations. CAT was more sensitive than SOD or POD. The diversity indices and composition at phylum level of the gut microbiota showed little change. However, the operational taxonomic units in the gut decreased in the presence of norfloxacin. The relative abundance of Wolbachia, the predominant bacterial genus in the gut, decreased significantly with increasing soil norfloxacin concentration. Wolbachia may therefore be a promising bioindicator in the assessment of norfloxacin pollution of soils at environmental concentrations.}, } @article {pmid34028446, year = {2021}, author = {Li, I and Mak, KW and Wong, J and Tan, CH}, title = {Using the Fluorescent Dye, Rhodamine B, to Study Mating Competitiveness in Male Aedes aegypti Mosquitoes.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {171}, pages = {}, doi = {10.3791/62432}, pmid = {34028446}, issn = {1940-087X}, mesh = {*Aedes ; Animals ; Female ; Fluorescent Dyes ; Male ; Mosquito Control ; Rhodamines ; Sexual Behavior, Animal ; *Wolbachia ; }, abstract = {The success of sterile or incompatible insect technique-based population suppression programs depends on the ability of released males to compete for wild-type females and induce sterility in the target population. Hence, laboratory assessment of male mating competitiveness is essential for evaluating the release strain's fitness before field release. Conventionally, such an assay is performed by determining the proportion of viable eggs produced by the females after being simultaneously exposed to two sets of males (wild-type and release strains) for copulation. However, this process is time-consuming and laborious due to the need to first blood-feed the females for egg production and then hatch and enumerate the hatched eggs to determine egg viability. Moreover, this method cannot discern the degree of competitiveness between two sterile or Wolbachia-infected mosquito lines as wild-type female mosquitoes will only produce non-viable eggs upon mating with both. To circumvent these limitations, this paper describes a more direct method of measuring male mosquito mating competitiveness in laboratory settings using the fluorescent dye, rhodamine B (RhB), which can be used to mark males by feeding them in sucrose solution containing RhB. After the mating assay, the presence of fluorescing sperms in the spermathecae of a female can be used to determine her mating partner. This method is cost-effective, reduces the experimental time by 90% and allows comparison of mating fitness between two sterile or Wolbachia-infected lines.}, } @article {pmid34026032, year = {2021}, author = {Sinotte, VM and Conlon, BH and Seibel, E and Schwitalla, JW and de Beer, ZW and Poulsen, M and Bos, N}, title = {Female-biased sex allocation and lack of inbreeding avoidance in Cubitermes termites.}, journal = {Ecology and evolution}, volume = {11}, number = {10}, pages = {5598-5605}, pmid = {34026032}, issn = {2045-7758}, abstract = {Sexually reproducing organisms face a strong selective pressure to find a mate and ensure reproduction. An important criterion during mate-selection is to avoid closely related individuals and subsequent potential fitness costs of resulting inbred offspring. Inbreeding avoidance can be active through kin recognition during mate choice, or passive through differential male and female-biased sex ratios, which effectively prevents sib-mating. In addition, sex allocation, or the resources allotted to male and female offspring, can impact mating and reproductive success. Here, we investigate mate choice, sex ratios, and sex allocation in dispersing reproductives (alates) from colonies of the termite Cubitermes tenuiceps. Termites have a short time to select a mate for life, which should intensify any fitness consequences of inbreeding. However, alates did not actively avoid inbreeding through mate choice via kin recognition based on genetic or environmental cues. Furthermore, the majority of colonies exhibited a female-biased sex ratio, and none exhibited a male-bias, indicating that differential bias does not reduce inbreeding. Sex allocation was generally female-biased, as females also were heavier, but the potential fitness effect of this costly strategy remains unclear. The bacterium Wolbachia, known in other insects to parasitically distort sex allocation toward females, was present within all alates. While Wolbachia is commonly associated with termites, parasitism has yet to be demonstrated, warranting further study of the nature of the symbiosis. Both the apparent lack of inbreeding avoidance and potential maladaptive sex allocation implies possible negative effects on mating and fitness.}, } @article {pmid34020585, year = {2021}, author = {Lucek, K and Bouaouina, S and Jospin, A and Grill, A and de Vos, JM}, title = {Prevalence and relationship of endosymbiotic Wolbachia in the butterfly genus Erebia.}, journal = {BMC ecology and evolution}, volume = {21}, number = {1}, pages = {95}, pmid = {34020585}, issn = {2730-7182}, mesh = {Animals ; *Butterflies ; Phylogeny ; Prevalence ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Wolbachia is an endosymbiont common to most invertebrates, which can have significant evolutionary implications for its host species by acting as a barrier to gene flow. Despite the importance of Wolbachia, still little is known about its prevalence and diversification pattern among closely related host species. Wolbachia strains may phylogenetically coevolve with their hosts, unless horizontal host-switches are particularly common. We address these issues in the genus Erebia, one of the most diverse Palearctic butterfly genera.

RESULTS: We sequenced the Wolbachia genome from a strain infecting Erebia cassioides and showed that it belongs to the Wolbachia supergroup B, capable of infecting arthropods from different taxonomic orders. The prevalence of Wolbachia across 13 closely related Erebia host species based on extensive population-level genetic data revealed that multiple Wolbachia strains jointly infect all investigated taxa, but with varying prevalence. Finally, the phylogenetic relationships of Wolbachia strains are in some cases significantly associated to that of their hosts, especially among the most closely related Erebia species, demonstrating mixed evidence for phylogenetic coevolution.

CONCLUSIONS: Closely related host species can be infected by closely related Wolbachia strains, evidencing some phylogenetic coevolution, but the actual pattern of infection more often reflects historical or contemporary geographic proximity among host species. Multiple processes, including survival in distinct glacial refugia, recent host shifts in sympatry, and a loss of Wolbachia during postglacial range expansion seem to have jointly shaped the complex interactions between Wolbachia evolution and the diversification of its host among our studied Erebia species.}, } @article {pmid34015229, year = {2021}, author = {Mioduchowska, M and Nitkiewicz, B and Roszkowska, M and Kačarević, U and Madanecki, P and Pinceel, T and Namiotko, T and Gołdyn, B and Kaczmarek, Ł}, title = {Taxonomic classification of the bacterial endosymbiont Wolbachia based on next-generation sequencing: is there molecular evidence for its presence in tardigrades?.}, journal = {Genome}, volume = {64}, number = {10}, pages = {951-958}, doi = {10.1139/gen-2020-0036}, pmid = {34015229}, issn = {1480-3321}, mesh = {Animals ; High-Throughput Nucleotide Sequencing ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Tardigrada/*microbiology ; *Wolbachia/classification ; }, abstract = {We used high-throughput sequencing of 16S rRNA to test whether tardigrade species are infected with Wolbachia parasites. We applied SILVA and Greengenes databases that allowed taxonomic classification of bacterial sequences to OTUs. The results obtained from both databases differed considerably in the number of OTUs, and only the Greengenes database allowed identification of Wolbachia (infection was also supported by comparison of sequences to NCBI database). The putative bacterial endosymbiont Wolbachia was discovered only in adult eutardigrades, while bacteria identified down to the order Rickettsiales were detected in both eutardigrade eggs and adult specimens. Nevertheless, the frequency of Wolbachia in the bacterial communities of the studied eutardigrades was low. Similarly, in our positive control, i.e., a fairy shrimp Streptocephalus cafer, which was found to be infected with Wolbachia in our previous study using Sanger sequencing, only the Rickettsiales were detected. We also carried out phylogenetic reconstruction using Wolbachia sequences from the SILVA and Greengenes databases, Alphaproteobacteria putative endosymbionts and Rickettsiales OTUs obtained in previous studies on the microbial community of tardigrades, and Rickettsiales and Wolbachia OTUs obtained in the current study. Our discovery of Wolbachia in tardigrades can fuel new research to uncover the specifics of this interaction.}, } @article {pmid34013860, year = {2021}, author = {Huebl, L and Tappe, D and Giese, M and Mempel, S and Tannich, E and Kreuels, B and Ramharter, M and Veletzky, L and Jochum, J}, title = {Recurrent Swelling and Microfilaremia Caused by Dirofilaria repens Infection after Travel to India.}, journal = {Emerging infectious diseases}, volume = {27}, number = {6}, pages = {1701-1704}, pmid = {34013860}, issn = {1080-6059}, mesh = {Animals ; *Dirofilaria repens ; *Dirofilariasis ; Germany ; Humans ; India ; Travel ; }, abstract = {Human subcutaneous dirofilariasis is an emerging mosquitoborne zoonosis. A traveler returning to Germany from India experienced Dirofilaria infection with concomitant microfilaremia. Molecular analysis indicated Dirofilaria repens nematodes of an Asian genotype. Microfilaremia showed no clear periodicity. Presence of Wolbachia endosymbionts enabled successful treatment with doxycycline.}, } @article {pmid34010697, year = {2021}, author = {Song, G and Chen, F and Chen, S and Ye, S}, title = {Polysaccharides from Premna microphylla turcz ameliorate inflammation via the enhancement of intestinal resistance in host.}, journal = {Journal of ethnopharmacology}, volume = {276}, number = {}, pages = {114208}, doi = {10.1016/j.jep.2021.114208}, pmid = {34010697}, issn = {1872-7573}, mesh = {Animals ; Autophagy/drug effects/immunology ; Disease Models, Animal ; Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster ; Epithelial Cells/drug effects ; Female ; Gastrointestinal Microbiome/drug effects/physiology ; Inflammation/chemically induced/genetics/mortality/*prevention & control ; Intestines/*drug effects/*immunology/microbiology/pathology ; Lamiaceae/*chemistry ; Metabolic Networks and Pathways ; Plant Extracts/*pharmacology/therapeutic use ; Polysaccharides/*pharmacology/therapeutic use ; Pore Forming Cytotoxic Proteins/genetics/metabolism ; Principal Component Analysis ; Protective Agents/*pharmacology/therapeutic use ; Sodium Dodecyl Sulfate/toxicity ; Survival Rate ; TOR Serine-Threonine Kinases/genetics/metabolism ; }, abstract = {Premna microphylla turcz is traditionally used as a folk remedy. Its roots, stems and leaves can be invoked as medicines, which have the functions of detoxification, swelling and hemostasis. It belongs to the Premna in the Verbenaceae and is mainly distributed in the mountains of southeastern China. However, there are few reports of in-depth studies on the anti-inflammatory effects of polysaccharide, which was the main component in Premna microphylla turcz.

MATERIALS AND METHODS: The flies were fed with standard corn flour-yeast medium to cause inflammation by sodium lauryl sulfate (SDS). The treatment group contained Premna microphylla turcz polysaccharide (pPMTLs) extract. The survival rate was obtained by feeding a vial containing five layers of filter paper, which was infiltrated with the 5% sucrose solution contaminated with SDS or SDS polysaccharide. The microvilli and nucleus of the midgut epithelial cells of different treatments were observed by transmission electron microscope, and the expression of inflammation-related genes was detected by real-time quantitative PCR (qRT-PCR). Finally, 16S rDNA analysis was conducted on the differences in the composition of the intestinal microbes of Drosophila.

RESULTS: In the current study, we showed that pPMTLs significantly prolonged the life span of SDS-inflamed flies from 5 days to 6 days. And pPMTLs reduced the rupture of microvilli in the midgut and restored the nuclear structure. In addition, pPMTLs significantly improved expression level of immune-related genes in Inflammation Drosophila especially the defensin (4.32 ± 0.75 vs 9.97 ± 0.52 SDS-polysaccharide group: SDS group, p < 0.001). The analysis of intestinal microbiota showed that pPMTLs decreased the relative abundance of Raoultella while Wolbachia increased (p < 0.05).

CONCLUSIONS: Collectively, our results revealed the potential application of pPMTLs in enhancing inflammation defense, which would be enormous significance for the inflammation-related disorders treatment.}, } @article {pmid34003269, year = {2021}, author = {Baião, GC and Janice, J and Galinou, M and Klasson, L}, title = {Comparative Genomics Reveals Factors Associated with Phenotypic Expression of Wolbachia.}, journal = {Genome biology and evolution}, volume = {13}, number = {7}, pages = {}, pmid = {34003269}, issn = {1759-6653}, mesh = {Animals ; Cytoplasm/genetics ; Female ; Genomics ; Male ; Phenotype ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a widespread, vertically transmitted bacterial endosymbiont known for manipulating arthropod reproduction. Its most common form of reproductive manipulation is cytoplasmic incompatibility (CI), observed when a modification in the male sperm leads to embryonic lethality unless a compatible rescue factor is present in the female egg. CI attracts scientific attention due to its implications for host speciation and in the use of Wolbachia for controlling vector-borne diseases. However, our understanding of CI is complicated by the complexity of the phenotype, whose expression depends on both symbiont and host factors. In the present study, we perform a comparative analysis of nine complete Wolbachia genomes with known CI properties in the same genetic host background, Drosophila simulans STC. We describe genetic differences between closely related strains and uncover evidence that phages and other mobile elements contribute to the rapid evolution of both genomes and phenotypes of Wolbachia. Additionally, we identify both known and novel genes associated with the modification and rescue functions of CI. We combine our observations with published phenotypic information and discuss how variability in cif genes, novel CI-associated genes, and Wolbachia titer might contribute to poorly understood aspects of CI such as strength and bidirectional incompatibility. We speculate that high titer CI strains could be better at invading new hosts already infected with a CI Wolbachia, due to a higher rescue potential, and suggest that titer might thus be a relevant parameter to consider for future strategies using CI Wolbachia in biological control.}, } @article {pmid33976301, year = {2021}, author = {Gesto, JSM and Ribeiro, GS and Rocha, MN and Dias, FBS and Peixoto, J and Carvalho, FD and Pereira, TN and Moreira, LA}, title = {Reduced competence to arboviruses following the sustainable invasion of Wolbachia into native Aedes aegypti from Southeastern Brazil.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {10039}, pmid = {33976301}, issn = {2045-2322}, mesh = {Aedes/*virology ; Animals ; Arboviruses/*physiology ; Brazil ; Dengue Virus/isolation & purification ; Female ; Mosquito Vectors/*virology ; Pest Control, Biological/methods/*statistics & numerical data ; *Wolbachia ; Zika Virus/isolation & purification ; }, abstract = {Field release of Wolbachia-infected Aedes aegypti has emerged as a promising solution to manage the transmission of dengue, Zika and chikungunya in endemic areas across the globe. Through an efficient self-dispersing mechanism, and the ability to induce virus-blocking properties, Wolbachia offers an unmatched potential to gradually modify wild Ae. aegypti populations turning them unsuitable disease vectors. Here we describe a proof-of-concept field trial carried out in a small community of Niterói, greater Rio de Janeiro, Brazil. Following the release of Wolbachia-infected eggs, we report here a successful invasion and long-term establishment of the bacterium across the territory, as denoted by stable high-infection indexes (> 80%). We have also demonstrated that refractoriness to dengue and Zika viruses, either thorough oral-feeding or intra-thoracic saliva challenging assays, was maintained over the adaptation to the natural environment of Southeastern Brazil. These findings further support Wolbachia's ability to invade local Ae. aegypti populations and impair disease transmission, and will pave the way for future epidemiological and economic impact assessments.}, } @article {pmid33969432, year = {2022}, author = {Nasehi, SF and Fathipour, Y and Asgari, S and Mehrabadi, M}, title = {Environmental Temperature, but Not Male Age, Affects Wolbachia and Prophage WO Thereby Modulating Cytoplasmic Incompatibility in the Parasitoid Wasp, Habrobracon Hebetor.}, journal = {Microbial ecology}, volume = {83}, number = {2}, pages = {482-491}, pmid = {33969432}, issn = {1432-184X}, support = {98008582//Iran National Science Foundation/ ; }, mesh = {Animals ; Cytoplasm/metabolism ; Female ; Male ; Prophages/genetics ; Temperature ; *Wasps/microbiology ; *Wolbachia/genetics/metabolism ; }, abstract = {Wolbachia is an endosymbiotic bacterium found in many species of arthropods and manipulates its host reproduction. Cytoplasmic incompatibility (CI) is one of the most common manipulations that is induced when an uninfected female mates with a Wolbachia-infected male. The CI factors (cifA and cifB genes) are encoded by phage WO that naturally infects Wolbachia. Here, we questioned whether an environmental factor (temperature) or host factor (male age) affected the strength of the CI phenotype in the ectoparasitoid wasp, Habrobracon hebetor. We found that temperature, but not male age, results in reduced CI penetrance. Consistent with these results, we also found that the expression of the cif CI factors decreased in temperature-exposed males but was consistent across aging male wasps. Similar to studies of other insect systems, cifA showed a higher expression level than cifB, and male hosts showed increased cif expression relative to females. Our results suggest that prophage WO is present in the Wolbachia-infected wasps and expression of cif genes contributes to the induction of CI in this insect. It seems that male aging has no effect on the intensity of CI; however, temperature affects Wolbachia and prophage WO titers as well as expression levels of cif genes, which modulate the CI level.}, } @article {pmid33962669, year = {2021}, author = {Manoj, RRS and Latrofa, MS and Epis, S and Otranto, D}, title = {Wolbachia: endosymbiont of onchocercid nematodes and their vectors.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {245}, pmid = {33962669}, issn = {1756-3305}, mesh = {Animals ; Arthropods/microbiology/physiology ; Host-Pathogen Interactions ; Nematoda/growth & development/immunology/*microbiology ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Wolbachia is an obligate intracellular maternally transmitted, gram-negative bacterium which forms a spectrum of endosymbiotic relationships from parasitism to obligatory mutualism in a wide range of arthropods and onchocercid nematodes, respectively. In arthropods Wolbachia produces reproductive manipulations such as male killing, feminization, parthenogenesis and cytoplasmic incompatibility for its propagation and provides an additional fitness benefit for the host to protect against pathogens, whilst in onchocercid nematodes, apart from the mutual metabolic dependence, this bacterium is involved in moulting, embryogenesis, growth and survival of the host.

METHODS: This review details the molecular data of Wolbachia and its effect on host biology, immunity, ecology and evolution, reproduction, endosymbiont-based treatment and control strategies exploited for filariasis. Relevant peer-reviewed scientic papers available in various authenticated scientific data bases were considered while writing the review.

CONCLUSIONS: The information presented provides an overview on Wolbachia biology and its use in the control and/or treatment of vectors, onchocercid nematodes and viral diseases of medical and veterinary importance. This offers the development of new approaches for the control of a variety of vector-borne diseases.}, } @article {pmid33958611, year = {2021}, author = {Tuda, M and Iwase, SI and Kébé, K and Haran, J and Skuhrovec, J and Sanaei, E and Tsuji, N and Podlussány, A and Merkl, O and El-Heneidy, AH and Morimoto, K}, title = {Diversification, selective sweep, and body size in the invasive Palearctic alfalfa weevil infected with Wolbachia.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {9664}, pmid = {33958611}, issn = {2045-2322}, mesh = {Animals ; Asia ; Body Size ; Europe ; Female ; Genetic Variation/genetics ; Haplotypes/genetics ; Introduced Species ; Male ; Mitochondria/genetics ; Phylogeny ; Phylogeography ; Weevils/genetics/*microbiology ; *Wolbachia ; }, abstract = {The alfalfa weevil Hypera postica, native to the Western Palearctic, is an invasive legume pest with two divergent mitochondrial clades in its invading regions, the Western clade and the Eastern/Egyptian clade. However, knowledge regarding the native populations is limited. The Western clade is infected with the endosymbiotic bacteria Wolbachia that cause cytoplasmic incompatibility in host weevils. Our aim was to elucidate the spatial genetic structure of this insect and the effect of Wolbachia on its population diversity. We analyzed two mitochondrial and two nuclear genes of the weevil from its native ranges. The Western clade was distributed in western/central Europe, whereas the Eastern/Egyptian clade was distributed from the Mediterranean basin to central Asia. Intermediate mitotypes were found from the Balkans to central Asia. Most Western clade individuals in western Europe were infected with an identical Wolbachia strain. Mitochondrial genetic diversity of the infected individuals was minimal. The infected clades demonstrated a higher nonsynonymous/synonymous substitution rate ratio than the uninfected clades, suggesting a higher fixation of nonsynonymous mutations due to a selective sweep by Wolbachia. Trans-Mediterranean and within-European dispersal routes were supported. We suggest that the ancestral populations diversified by geographic isolation due to glaciations and that the diversity was reduced in the west by a recent Wolbachia-driven sweep(s). The intermediate clade exhibited a body size and host plant that differed from the other clades. Pros and cons of the possible use of infected-clade males to control uninfected populations are discussed.}, } @article {pmid33955029, year = {2021}, author = {Pilgrim, J and Siozios, S and Baylis, M and Venter, G and Garros, C and Hurst, GDD}, title = {Identifying potential candidate Culicoides spp. for the study of interactions with Candidatus Cardinium hertigii.}, journal = {Medical and veterinary entomology}, volume = {35}, number = {3}, pages = {501-506}, doi = {10.1111/mve.12517}, pmid = {33955029}, issn = {1365-2915}, support = {BB/M011186/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Bacteroidetes ; *Ceratopogonidae ; Mosquito Vectors ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Culicoides biting midges (Diptera: Ceratopogonidae) are vectors responsible for the transmission of several viruses of veterinary importance. Previous screens of Culicoides have described the presence of the endosymbiont Candidatus Cardinium hertigii (Bacteroidetes). However, any impacts of this microbe on vectorial capacity, akin to those conferred by Wolbachia in mosquitoes, are yet to be uncovered and await a suitable system to study Cardinium-midge interactions. To identify potential candidate species to investigate these interactions, accurate knowledge of the distribution of the endosymbiont within Culicoides populations is needed. We used conventional and nested PCR assays to screen Cardinium infection in 337 individuals of 25 Culicoides species from both Palearctic and Afrotropical regions. Infections were observed in several vector species including C. imicola and the Pulicaris complex (C. pulicaris, C. bysta, C. newsteadi and C. punctatus) with varying prevalence. Phylogenetic analysis based on the Gyrase B gene grouped all new isolates within 'group C' of the genus, a clade that has to date been exclusively described in Culicoides. Through a comparison of our results with previous screens, we suggest C. imicola and C. sonorensis represent good candidates for onward study of Cardinium-midge interactions.}, } @article {pmid33948978, year = {2021}, author = {Manoj, RRS and Latrofa, MS and Cavalera, MA and Mendoza-Roldan, JA and Maia, C and Otranto, D}, title = {Molecular detection of zoonotic filarioids in Culex spp. from Portugal.}, journal = {Medical and veterinary entomology}, volume = {35}, number = {3}, pages = {468-477}, pmid = {33948978}, issn = {1365-2915}, mesh = {*Aedes ; Animals ; *Culex ; *Dirofilaria immitis ; Mosquito Vectors ; Portugal ; }, abstract = {To investigate the role of dipterans in the transmission of Onchocerca lupi and other zoonotic filarioids, samples were collected from different sites in Algarve, southern Portugal, morphologically identified and molecularly tested for filarioids. Culex sp. (72.8%) represented the predominant genus followed by Culicoides sp. (11.8%), Ochlerotatus sp. (9.7%), Culiseta sp. (4.5%), Aedes sp. (0.9%) and Anopheles sp. (0.3%). Nineteen (2.8%) specimens scored positive for filarioids, with Culex pipiens quinquefasciatus (2%) positive for Dirofilaria immitis (1.4%), Dirofilaria repens, Acanthocheilonema reconditum, Onchocerca lupi, unidentified species of Filarioidea (0.2%, each) and Onchocercidae (0.6%). Additionally, Culiseta longiareolata (6.5%), Ochlerotatus caspius (3%) and Culex laticinctus (0.2%) scored positive for unidentified Onchocercidae, A. reconditum and for O. lupi, respectively. This is the first report of the occurrence of DNA of O. lupi, D. repens and A. reconditum in Culex spp. in Portugal. Information regarding the vectors and the pathogens they transmit may help to adopt proper prophylactic and control measures.}, } @article {pmid33947218, year = {2021}, author = {Hague, MTJ and Woods, HA and Cooper, BS}, title = {Pervasive effects of Wolbachia on host activity.}, journal = {Biology letters}, volume = {17}, number = {5}, pages = {20210052}, pmid = {33947218}, issn = {1744-957X}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila ; Locomotion ; Reproduction ; Symbiosis ; *Wolbachia ; }, abstract = {Heritable symbionts have diverse effects on the physiology, reproduction and fitness of their hosts. Maternally transmitted Wolbachia are one of the most common endosymbionts in nature, infecting about half of all insect species. We test the hypothesis that Wolbachia alter host behaviour by assessing the effects of 14 different Wolbachia strains on the locomotor activity of nine Drosophila host species. We find that Wolbachia alter the activity of six different host genotypes, including all hosts in our assay infected with wRi-like Wolbachia strains (wRi, wSuz and wAur), which have rapidly spread among Drosophila species in about the last 14 000 years. While Wolbachia effects on host activity were common, the direction of these effects varied unpredictably and sometimes depended on host sex. We hypothesize that the prominent effects of wRi-like Wolbachia may be explained by patterns of Wolbachia titre and localization within host somatic tissues, particularly in the central nervous system. Our findings support the view that Wolbachia have wide-ranging effects on host behaviour. The fitness consequences of these behavioural modifications are important for understanding the evolution of host-symbiont interactions, including how Wolbachia spread within host populations.}, } @article {pmid33945798, year = {2021}, author = {Kaur, R and Shropshire, JD and Cross, KL and Leigh, B and Mansueto, AJ and Stewart, V and Bordenstein, SR and Bordenstein, SR}, title = {Living in the endosymbiotic world of Wolbachia: A centennial review.}, journal = {Cell host & microbe}, volume = {29}, number = {6}, pages = {879-893}, pmid = {33945798}, issn = {1934-6069}, support = {F32 AI140694/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/physiology ; Biological Evolution ; Feminization ; *Host Microbial Interactions ; Host Specificity ; Humans ; Male ; Phenotype ; Phylogeny ; Preventive Medicine ; *Symbiosis ; Wolbachia/*cytology/*physiology/*virology ; }, abstract = {The most widespread intracellular bacteria in the animal kingdom are maternally inherited endosymbionts of the genus Wolbachia. Their prevalence in arthropods and nematodes worldwide and stunning arsenal of parasitic and mutualistic adaptations make these bacteria a biological archetype for basic studies of symbiosis and applied outcomes for curbing human and agricultural diseases. Here, we conduct a summative, centennial analysis of living in the Wolbachia world. We synthesize literature on Wolbachia's host range, phylogenetic diversity, genomics, cell biology, and applications to filarial, arboviral, and agricultural diseases. We also review the mobilome of Wolbachia including phage WO and its essentiality to hallmark reproductive phenotypes in arthropods. Finally, the Wolbachia system is an exemplar for discovery-based science education using biodiversity, biotechnology, and bioinformatics lessons. As we approach a century of Wolbachia research, the interdisciplinary science of this symbiosis stands as a model for consolidating and teaching the integrative rules of endosymbiotic life.}, } @article {pmid33925275, year = {2021}, author = {Pereira, TN and Carvalho, FD and Rugani, JN and de Carvalho, VR and Jarusevicius, J and Souza-Neto, JA and Moreira, LA}, title = {Mayaro Virus: The Potential Role of Microbiota and Wolbachia.}, journal = {Pathogens (Basel, Switzerland)}, volume = {10}, number = {5}, pages = {}, pmid = {33925275}, issn = {2076-0817}, abstract = {The Mayaro virus (MAYV) is an arbovirus that circulates mainly in tropical forests or rural areas in Latin America and is transmitted mainly by Haemagogus mosquitoes. The objective of this study was to evaluate the vector competence, microbiome, and the presence of Wolbachia in three Aedes albopictus populations infected with MAYV. The vector competence was assessed based on viral infection and transmission by RT-qPCR. In addition, the microbiome was evaluated by amplification of the 16S rRNA V4 region and PCR to detect the presence of Wolbachia (strain wAlbA/wAlbB). Our results show that all three populations were susceptible to MAYV infection. The potential transmission of the MAYV was consistent in all populations of naïve mosquitoes injected (more than 50%). The microbiome analysis revealed 118 OTUs (operational taxonomic unit) from the three populations, 8 phyla, 15 classes, 26 orders, 35 families, 65 genera, and 53 species. All populations had Pseudomonas and Wolbachia as predominant genera. There was no difference between the variables for MAYV and Wolbachia (wAlbA or wAlbB) in the abdomen. However, in the head + thorax samples at 14 dpi, there was a difference between the two populations, indicating a possible correlation between the presence of Wolbachia (wAlbB) and infection. Overall, we show evidence that Ae. albopictus displays significant infection and transmission competence for the MAYV in the laboratory, and its bacterial microbiota play an important role in the host, mainly the strains of Wolbachia. The influence of the intestinal microbiota of Ae. albopictus is poorly known, and a better understanding of these interactions would open new perspectives for disease control through the manipulation of microbial communities. The exact contribution of this mosquito species to the transmission of the MAYV in the field remains to be confirmed.}, } @article {pmid33922427, year = {2021}, author = {Guad, RM and Wu, YS and Aung, YN and Sekaran, SD and Wilke, ABB and Low, WY and Sim, MS and Carandang, RR and Jeffree, MS and Taherdoost, H and Sunggip, C and Lin, CLS and Murugaiah, C and Subramaniyan, V and Azizan, N}, title = {Different Domains of Dengue Research in Malaysia: A Systematic Review and Meta-Analysis of Questionnaire-Based Studies.}, journal = {International journal of environmental research and public health}, volume = {18}, number = {9}, pages = {}, pmid = {33922427}, issn = {1660-4601}, mesh = {Child ; *Dengue/epidemiology/prevention & control ; *Health Knowledge, Attitudes, Practice ; Humans ; Malaysia/epidemiology ; Reproducibility of Results ; Surveys and Questionnaires ; }, abstract = {This review provided a systematic overview of the questionnaire-related dengue studies conducted in Malaysia and evaluated their reliability and validity used in the questionnaires. An extensive literature search was conducted using various electronic databases, including PubMed, EMBASE, Medline, and ScienceDirect. Systematic reviews and meta-analysis (PRISMA) were selected as the preferred item reporting method. Out of 88 identified dengue-related, 57 published from 2000 to April 2020 met the inclusion criteria and were included. Based on the meta-analysis, a poor mean score was obtained for knowledge (49%), attitude (44%), and preventive practice (55%). The study showed that the level of knowledge on cardinal signs and modes of transmission for dengue virus were highest among health care workers, followed by students (international and local) and lastly community residents. In treatment-seeking behaviours, only half of the respondents (50.8%) would send their child to the nearest health clinics or hospitals when a child became restless or lethargic. The acceptance rate for dengue vaccine, bacteria (Wolbachia), as a vector for dengue control and self-test diagnostic kit for dengue showed considerably high (88.4%, 70%, and 44.8%, respectively). Health belief model (HBM) constructs, such as perceived barriers, perceived severity, perceived susceptibility, self-efficacy, and perceived benefit influence prevention practices. Lastly, only 23 articles (40.3%) had piloted or pretested the questionnaire before surveying, in which three reported Cronbach's alpha coefficient (0.70-0.90). A need for active participation of communities and healthcare personnel, promotion of awareness, and safe complementary medicines, as well as assessment of psychometric properties of questionnaire use in dengue surveys in Malaysia, in order for assessing dengue reliably and valid.}, } @article {pmid33920445, year = {2021}, author = {Luu, L and Palomar, AM and Farrington, G and Schilling, AK and Premchand-Branker, S and McGarry, J and Makepeace, BL and Meredith, A and Bell-Sakyi, L}, title = {Bacterial Pathogens and Symbionts Harboured by Ixodes ricinus Ticks Parasitising Red Squirrels in the United Kingdom.}, journal = {Pathogens (Basel, Switzerland)}, volume = {10}, number = {4}, pages = {}, pmid = {33920445}, issn = {2076-0817}, support = {BB/N023889/2/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; Becas de estancia en el extranjero//Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica/ ; BBS/E/1/00001741/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, abstract = {Red squirrels (Sciurus vulgaris) are native to most of Eurasia; in much of the United Kingdom, they have been supplanted by the non-native grey squirrel, and are considered an endangered species. Very little is known about the range of tick-borne pathogens to which UK red squirrels are exposed. As part of trap-and-release surveys examining prevalence of Mycobacterium spp. in red squirrel populations on two UK islands, Ixodes ricinus ticks were removed from squirrels and PCR screened for Borrelia spp., intracellular arthropod-borne bacteria and the parasitic wasp Ixodiphagus hookeri. At both sites, the most commonly encountered tick-transmitted bacterium was Borrelia burgdorferi sensu lato (overall minimum prevalence 12.7%), followed by Anaplasma phagocytophilum (overall minimum prevalence 1.6%). Single ticks infected with Spiroplasma were found at both sites, and single ticks infected with Borrelia miyamotoi or an Ehrlichia sp. at one site. Ticks harbouring Wolbachia (overall minimum prevalence 15.2%) were all positive for I. hookeri. Our study shows that UK red squirrels are potentially exposed to a variety of bacterial pathogens via feeding ticks. The effects on the health and survival of this already vulnerable wildlife species are unknown, and further studies are needed to evaluate the threat posed to red squirrels by Borrelia and other tick-borne pathogens.}, } @article {pmid33916889, year = {2021}, author = {Liu, G and Zheng, X and Long, H and Rao, Z and Cao, L and Han, R}, title = {Gut Bacterial and Fungal Communities of the Wild and Laboratory-Reared Thitarodes Larvae, Host of the Chinese Medicinal Fungus Ophiocordyceps sinensis on Tibetan Plateau.}, journal = {Insects}, volume = {12}, number = {4}, pages = {}, pmid = {33916889}, issn = {2075-4450}, abstract = {By employing a culture-dependent and -independent 16S rRNA and ITS gene high-throughput sequencing analyses, comprehensive information was obtained on the gut bacterial and fungal communities in the ghost moth larvae of three different geographic locations from high-altitude on Tibet plateau and from low-altitude laboratory. Twenty-six culturable bacterial species belonging to 21 genera and 14 fungal species belonging to 12 genera were identified from six populations by culture-dependent method. Carnobacterium maltaromaticum was the most abundant bacterial species from both the wild and laboratory-reared larvae. The most abundant OTUs in the wild ghost moth populations were Carnobacteriaceae, Enterobacteriaceae for bacteria, and Ascomycota and Basidiomycota for fungi. Larval microbial communities of the wild ghost moth from different geographic locations were not significantly different from each other but significant difference in larval microbial community was detected between the wild and laboratory-reared ghost moth. The larval gut of the wild ghost moth was dominated by the culturable Carnobacterium. However, that of the laboratory-reared ghost moth exhibited significantly abundant Wolbachia, Rhizobium, Serratia, Pseudomonas, and Flavobacterium. Furthermore, the larval gut of the wild ghost moth had a significantly higher abundance of Ophiocordyceps but lower abundance of Candida and Aspergillus than that of the laboratory-reared ghost moth.}, } @article {pmid33912718, year = {2021}, author = {Shemshadian, A and Vatandoost, H and Oshaghi, MA and Abai, MR and Djadid, ND and Karimian, F}, title = {Relationship between Wolbachia infection in Culex quinquefasciatus and its resistance to insecticide.}, journal = {Heliyon}, volume = {7}, number = {4}, pages = {e06749}, pmid = {33912718}, issn = {2405-8440}, abstract = {Many studies have been done on the various factors affecting resistance to insecticides. The relationship between Wolbachia bacteria and resistance to insecticides is one of the factors that has attracted a lot of attentions. Wolbachia are obligatory intracellular endosymbionts that naturally occur in a wide range of arthropods and nematodes, including the mosquito Culex quinquefasciatus. Initially, the presence of bacteria was proved by molecular assays. Then the resistance level of this species was evaluated in adults against DDT 4.0% and deltamethrin 0.05% using the standard WHO guideline. After elimination of Wolbachia by tetracycline and its proof by molecular assays, the susceptibility tests were conducted again on uninfected line. Finally, the two lines were compared in terms of responding to insecticides. The findings indicated that there is no significant correlation between susceptibility of two lines in response to DDT 4.0% while they represented a significant correlation for deltamethrin (P =0.00). We propose that Wolbachia bacteria increase the susceptibility to deltamethrin but they show neutral effect on DDT susceptibility in Cx. quinquefasciatus. However, more studies on other vectors and insecticides still need to be done.}, } @article {pmid33906628, year = {2021}, author = {Knerer, G and Currie, CSM and Brailsford, SC}, title = {Reducing dengue fever cases at the lowest budget: a constrained optimization approach applied to Thailand.}, journal = {BMC public health}, volume = {21}, number = {1}, pages = {807}, pmid = {33906628}, issn = {1471-2458}, mesh = {Animals ; Child ; *Culicidae ; *Dengue/epidemiology/prevention & control ; Humans ; Mosquito Vectors ; Thailand/epidemiology ; *Wolbachia ; }, abstract = {BACKGROUND: With the challenges that dengue fever (DF) presents to healthcare systems and societies, public health officials must determine where best to allocate scarce resources and restricted budgets. Constrained optimization (CO) helps to address some of the acknowledged limitations of conventional health economic analyses and has typically been used to identify the optimal allocation of resources across interventions subject to a variety of constraints.

METHODS: A dynamic transmission model was developed to predict the number of dengue cases in Thailand at steady state. A CO was then applied to identify the optimal combination of interventions (release of Wolbachia-infected mosquitoes and paediatric vaccination) within the constraints of a fixed budget, set no higher than cost estimates of the current vector control programme, to minimize the number of dengue cases and disability-adjusted life years (DALYs) lost. Epidemiological, cost, and effectiveness data were informed by national data and the research literature. The time horizon was 10 years. Scenario analyses examined different disease management and intervention costs, budget constraints, vaccine efficacy, and optimization time horizon.

RESULTS: Under base-case budget constraints, the optimal coverage of the two interventions to minimize dengue incidence was predicted to be nearly equal (Wolbachia 50%; paediatric vaccination 49%) with corresponding coverages under lower bound (Wolbachia 54%; paediatric vaccination 10%) and upper bound (Wolbachia 67%; paediatric vaccination 100%) budget ceilings. Scenario analyses indicated that the most impactful situations related to the costs of Wolbachia and paediatric vaccination with decreases/ increases in costs of interventions demonstrating a direct correlation with coverage (increases/ decreases) of the respective control strategies under examination.

CONCLUSIONS: Determining the best investment strategy for dengue control requires the identification of the optimal mix of interventions to implement in order to maximize public health outcomes, often under fixed budget constraints. A CO model was developed with the objective of minimizing dengue cases (and DALYs lost) over a 10-year time horizon, within the constraints of the estimated budgets for vector control in the absence of vaccination and Wolbachia. The model provides a tool for developing estimates of optimal coverage of combined dengue control strategies that minimize dengue burden at the lowest budget.}, } @article {pmid33895462, year = {2021}, author = {Zurita, A and Benkacimi, L and El Karkouri, K and Cutillas, C and Parola, P and Laroche, M}, title = {New records of bacteria in different species of fleas from France and Spain.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {76}, number = {}, pages = {101648}, doi = {10.1016/j.cimid.2021.101648}, pmid = {33895462}, issn = {1878-1667}, mesh = {Animals ; *Bacteria/classification/genetics ; *Ctenocephalides/microbiology ; Europe ; *Flea Infestations/epidemiology/veterinary ; France ; *Siphonaptera/microbiology ; Spain/epidemiology ; }, abstract = {In this study, we assessed the presence of vector-borne microorganisms in different species of fleas collected from different hosts in diverse areas of South-Western Europe by molecular methods. A total of 319 fleas belonging to eight different species was tested for the presence of eight microorganisms. Wolbachia spp. endosymbionts were detected in Ctenocephalides felis, Pulex irritans, Archaeopsylla erinacei and Ctenophthalmus baeticus boisseauorum specimens. Rickettsia felis, an emerging pathogen, was detected in C. felis, A. erinacei and Ct. b. boisseauorum. Rickettsia typhi, the agent of murine typhus was detected for the first time in A. erinacei and Mycobacterium spp. were detected for the first time in fleas (C. felis, P. irritans and A. erinacei). Lastly, five different species of Bartonella were detected in fleas' DNA in this study, including a possible new bacterium belonging to this genus. With this study, we updated the knowledge of the flea-borne bacteria present in the South-West of Europe reinforcing the idea about the necessity to expand and increase the current knowledge on flea-borne pathogens.}, } @article {pmid33892579, year = {2021}, author = {Cardona-Salgado, D and Campo-Duarte, DE and Sepulveda-Salcedo, LS and Vasilieva, O and Svinin, M}, title = {Optimal release programs for dengue prevention using Aedes aegypti mosquitoes transinfected with wMel or wMelPop Wolbachia strains.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {18}, number = {3}, pages = {2952-2990}, doi = {10.3934/mbe.2021149}, pmid = {33892579}, issn = {1551-0018}, mesh = {*Aedes ; Animals ; *Dengue/prevention & control ; Female ; Humans ; Male ; Population Dynamics ; *Wolbachia ; }, abstract = {In this paper, we propose a dengue transmission model of SIR(S)-SI type that accounts for two sex-structured mosquito populations: the wild mosquitoes (males and females that are Wolbachia-free), and those deliberately infected with either wMel or wMelPop strain of Wolbachia. This epidemiological model has four possible outcomes: with or without Wolbachia and with or without dengue. To reach the desired outcome, with Wolbachia and without dengue, we employ the dynamic optimization approach and then design optimal programs for releasing Wolbachia-carrying male and female mosquitoes. Our discussion is focused on advantages and drawbacks of two Wolbachia strains, wMelPop and wMel, that are recommended for dengue prevention and control. On the one hand, the wMel strain guarantees a faster population replacement, ensures durable Wolbachia persistence in the wild mosquito population, and requiters fewer releases. On the other hand, the wMelPop strain displays better results for averting dengue infections in the human population.}, } @article {pmid33889137, year = {2021}, author = {Gabrieli, P and Caccia, S and Varotto-Boccazzi, I and Arnoldi, I and Barbieri, G and Comandatore, F and Epis, S}, title = {Mosquito Trilogy: Microbiota, Immunity and Pathogens, and Their Implications for the Control of Disease Transmission.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {630438}, pmid = {33889137}, issn = {1664-302X}, abstract = {In mosquitoes, the interaction between the gut microbiota, the immune system, and the pathogens that these insects transmit to humans and animals is regarded as a key component toward the development of control strategies, aimed at reducing the burden of severe diseases, such as malaria and dengue fever. Indeed, different microorganisms from the mosquito microbiota have been investigated for their ability to affect important traits of the biology of the host insect, related with its survival, development and reproduction. Furthermore, some microorganisms have been shown to modulate the immune response of mosquito females, significantly shaping their vector competence. Here, we will review current knowledge in this field, focusing on i) the complex interaction between the intestinal microbiota and mosquito females defenses, both in the gut and at humoral level; ii) how knowledge on these issues contributes to the development of novel and targeted strategies for the control of mosquito-borne diseases such as the use of paratransgenesis or taking advantage of the relationship between Wolbachia and mosquito hosts. We conclude by providing a brief overview of available knowledge on microbiota-immune system interplay in major insect vectors.}, } @article {pmid33882628, year = {2021}, author = {Wolfe, TM and Bruzzese, DJ and Klasson, L and Corretto, E and Lečić, S and Stauffer, C and Feder, JL and Schuler, H}, title = {Comparative genome sequencing reveals insights into the dynamics of Wolbachia in native and invasive cherry fruit flies.}, journal = {Molecular ecology}, volume = {30}, number = {23}, pages = {6259-6272}, pmid = {33882628}, issn = {1365-294X}, support = {P 31441/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Drosophila ; Multilocus Sequence Typing ; Symbiosis/genetics ; *Tephritidae/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a maternally inherited obligate endosymbiont that can induce a wide spectrum of effects in its host, ranging from mutualism to reproductive parasitism. At the genomic level, recombination within and between strains, transposable elements, and horizontal transfer of strains between host species make Wolbachia an evolutionarily dynamic bacterial system. The invasive cherry fruit fly Rhagoletis cingulata arrived in Europe from North America ~40 years ago, where it now co-occurs with the native cherry pest R. cerasi. This shared distribution has been proposed to have led to the horizontal transfer of different Wolbachia strains between the two species. To better understand transmission dynamics, we performed a comparative genome study of the strain wCin2 in its native United States and invasive European populations of R. cingulata with wCer2 in European R. cerasi. Previous multilocus sequence genotyping (MLST) of six genes implied that the source of wCer2 in R. cerasi was wCin2 from R. cingulata. However, we report genomic evidence discounting the recent horizontal transfer hypothesis for the origin of wCer2. Despite near identical sequences for the MLST markers, substantial sequence differences for other loci were found between wCer2 and wCin2, as well as structural rearrangements, and differences in prophage, repetitive element, gene content, and cytoplasmic incompatibility inducing genes. Our study highlights the need for whole-genome sequencing rather than relying on MLST markers for resolving Wolbachia strains and assessing their evolutionary dynamics.}, } @article {pmid33882275, year = {2021}, author = {Liu, XC and Li, ZX}, title = {Transmission of the wMel Wolbachia strain is modulated by its titre and by immune genes in Drosophila melanogaster (Wolbachia density and transmission).}, journal = {Journal of invertebrate pathology}, volume = {181}, number = {}, pages = {107591}, doi = {10.1016/j.jip.2021.107591}, pmid = {33882275}, issn = {1096-0805}, mesh = {Animals ; Drosophila melanogaster/genetics/*immunology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are common intracellular endosymbionts of arthropods, but the interactions between Wolbachia and arthropods are only partially understood. The fruit fly Drosophila melanogaster is a model insect for understanding Wolbachia-host interactions. Here the native wMel strain of D. melanogaster was isolated and then different initial titres of wMel were artificially transferred back into antibiotics-treated fruit flies. Our purpose was to examine the interactions between the injected wMel in a density gradient and the recipient host during trans-generational transmission. The results showed that the trans-generational transmission rates of wMel and titres of wMel exhibited a fluctuating trend over nine generations, and the titres of wMel displayed a similar fluctuating trans-generational trend. There was a significant positive correlation between the transmission rate and the titre of wMel. Reciprocal crossings between wMel-transinfected and uninfected fruit flies revealed that wMel could induce cytoplasmic incompatibility (CI) at different initial titres, but the intensity of CI was not significantly correlated with the initial titre of wMel. Quantitative PCR analysis showed that the immune genes Drsl5 and Spn38F displayed a significant transcriptional response to wMel transfection, with an obvious negative correlation with the titre of wMel at the 3rd and 4th generations. Furthermore, RNA interference-mediated knockdown of Drsl5 and Spn38F elicited a drastic increase in the titre of wMel. In combination, our study suggests that the trans-generational transmission of wMel is modulated by its density, and the immune genes are involved in the regulation of Wolbachia density.}, } @article {pmid33881802, year = {2022}, author = {Li, C and He, M and Cui, Y and Peng, Y and Liu, J and Yun, Y}, title = {Insights into the mechanism of shortened developmental duration and accelerated weight gain associated with Wolbachia infection in Hylyphantes graminicola.}, journal = {Integrative zoology}, volume = {17}, number = {3}, pages = {420-429}, doi = {10.1111/1749-4877.12550}, pmid = {33881802}, issn = {1749-4877}, support = {NSFC-31573236/31772420/31970406//National Natural Sciences Foundation of China/ ; 19CAS-TFE-3//CAS Key Laboratory of Tropical Forest Ecology, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences/ ; }, mesh = {Animals ; Body Weight ; Ecdysone ; *Insulins ; *Somatomedins ; *Spiders/genetics ; Weight Gain ; *Wolbachia/genetics ; }, abstract = {Wolbachia infection is known to affect host reproduction and development. To date, however, the underlying mechanism related to the effects of Wolbachia on host development has not yet been reported. Here, we compared the developmental duration and body weight in different instars of Wolbachia-positive (W[+]) and Wolbachia-negative (W[-]) spiders (Hylyphantes graminicola) and detected the relative expression levels of 6 insulin-related genes and 3 ecdysone-related genes using reverse transcription qPCR. Results showed that the developmental duration was significantly shortened in W[+] spiders compared with W[-] spiders. Furthermore, W[+] spiders were significantly heavier than W[-] spiders at the 3[rd] and 4[th] instars, although no significant differences in body weight were observed after maturity. We also found that the expression levels of insulin-like growth factor-2 mRNA-binding protein-1, insulin-degrading enzyme, and ecdysone-inducible protein-1 genes were significantly down-regulated in W[+] spiders compared with W[-] spiders, whereas the expression levels of insulin-like growth factor-binding protein-1, insulin-like peptide receptor, insulin receptor substrate 2-B, insulin-like, ecdysone-induced protein-2, and ecdysone receptor genes were significantly up-regulated in W[+] spiders. Our results suggest that Wolbachia may influence host development by affecting insulin and ecdysone signaling pathways.}, } @article {pmid33879316, year = {2021}, author = {Tain, LS and Sehlke, R and Meilenbrock, RL and Leech, T and Paulitz, J and Chokkalingam, M and Nagaraj, N and Grönke, S and Fröhlich, J and Atanassov, I and Mann, M and Beyer, A and Partridge, L}, title = {Tissue-specific modulation of gene expression in response to lowered insulin signalling in Drosophila.}, journal = {eLife}, volume = {10}, number = {}, pages = {}, pmid = {33879316}, issn = {2050-084X}, support = {268739/ERC_/European Research Council/International ; 0315893A-B//Bundesministerium für Bildung und Forschung/ ; FP7/2007-2013//FP7 Ideas: European Research Council/ ; }, mesh = {Animals ; Drosophila Proteins/*metabolism ; Drosophila melanogaster/*genetics/metabolism/microbiology ; Fat Body/metabolism ; Gene Expression Profiling ; Insulin/*physiology ; Proteome/*metabolism ; *Signal Transduction ; *Transcriptome ; Wolbachia/*physiology ; }, abstract = {Reduced activity of the insulin/IGF signalling network increases health during ageing in multiple species. Diverse and tissue-specific mechanisms drive the health improvement. Here, we performed tissue-specific transcriptional and proteomic profiling of long-lived Drosophila dilp2-3,5 mutants, and identified tissue-specific regulation of >3600 transcripts and >3700 proteins. Most expression changes were regulated post-transcriptionally in the fat body, and only in mutants infected with the endosymbiotic bacteria, Wolbachia pipientis, which increases their lifespan. Bioinformatic analysis identified reduced co-translational ER targeting of secreted and membrane-associated proteins and increased DNA damage/repair response proteins. Accordingly, age-related DNA damage and genome instability were lower in fat body of the mutant, and overexpression of a minichromosome maintenance protein subunit extended lifespan. Proteins involved in carbohydrate metabolism showed altered expression in the mutant intestine, and gut-specific overexpression of a lysosomal mannosidase increased autophagy, gut homeostasis, and lifespan. These processes are candidates for combatting ageing-related decline in other organisms.}, } @article {pmid33875732, year = {2021}, author = {Kwarteng, A and Asiedu, E and Sylverken, A and Larbi, A and Mubarik, Y and Apprey, C}, title = {In silico drug repurposing for filarial infection predicts nilotinib and paritaprevir as potential inhibitors of the Wolbachia 5'-aminolevulinic acid synthase.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {8455}, pmid = {33875732}, issn = {2045-2322}, mesh = {5-Aminolevulinate Synthetase/*antagonists & inhibitors ; Amino Acid Sequence ; *Computer Simulation ; Cyclopropanes/*pharmacology ; Drug Repositioning/*methods ; Enzyme Inhibitors/*pharmacology ; Humans ; Lactams, Macrocyclic/*pharmacology ; Proline/*analogs & derivatives/pharmacology ; Pyrimidines/*pharmacology ; Sequence Homology ; Sulfonamides/*pharmacology ; Wolbachia/*drug effects/enzymology/growth & development ; }, abstract = {Filarial infections affect millions of individuals and are responsible for some notorious disabilities. Current treatment options involve repeated mass drug administrations, which have been met with several challenges despite some successes. Administration of doxycycline, an anti-Wolbachia agent, has shown clinical effectiveness but has several limitations, including long treatment durations and contraindications. We describe the use of an in silico drug repurposing approach to screening a library of over 3200 FDA-approved medications against the filarial endosymbiont, Wolbachia. We target the enzyme which catalyzes the first step of heme biosynthesis in the Wolbachia. This presents an opportunity to inhibit heme synthesis, which leads to depriving the filarial worm of heme, resulting in a subsequent macrofilaricidal effect. High throughput virtual screening, molecular docking and molecular simulations with binding energy calculations led to the identification of paritaprevir and nilotinib as potential anti-Wolbachia agents. Having higher binding affinities to the catalytic pocket than the natural substrate, these drugs have the structural potential to bind and engage active site residues of the wolbachia 5'-Aminolevulinic Acid Synthase. We hereby propose paritaprevir and nilotinib for experimental validations as anti-Wolbachia agents.}, } @article {pmid33875727, year = {2021}, author = {Lim, L and Ab Majid, AH}, title = {Characterization of bacterial communities associated with blood-fed and starved tropical bed bugs, Cimex hemipterus (F.) (Hemiptera): a high throughput metabarcoding analysis.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {8465}, pmid = {33875727}, issn = {2045-2322}, mesh = {Animals ; Bacteria/*classification/genetics/*isolation & purification ; Bedbugs/*microbiology ; Blood/*microbiology ; DNA/analysis/*genetics ; DNA Barcoding, Taxonomic ; *Feeding Behavior ; *Starvation ; }, abstract = {With the development of new metagenomic techniques, the microbial community structure of common bed bugs, Cimex lectularius, is well-studied, while information regarding the constituents of the bacterial communities associated with tropical bed bugs, Cimex hemipterus, is lacking. In this study, the bacteria communities in the blood-fed and starved tropical bed bugs were analysed and characterized by amplifying the v3-v4 hypervariable region of the 16S rRNA gene region, followed by MiSeq Illumina sequencing. Across all samples, Proteobacteria made up more than 99% of the microbial community. An alpha-proteobacterium Wolbachia and gamma-proteobacterium, including Dickeya chrysanthemi and Pseudomonas, were the dominant OTUs at the genus level. Although the dominant OTUs of bacterial communities of blood-fed and starved bed bugs were the same, bacterial genera present in lower numbers were varied. The bacteria load in starved bed bugs was also higher than blood-fed bed bugs.}, } @article {pmid33868697, year = {2021}, author = {Jeffries, CL and Cansado-Utrilla, C and Beavogui, AH and Stica, C and Lama, EK and Kristan, M and Irish, SR and Walker, T}, title = {Evidence for natural hybridization and novel Wolbachia strain superinfections in the Anopheles gambiae complex from Guinea.}, journal = {Royal Society open science}, volume = {8}, number = {4}, pages = {202032}, pmid = {33868697}, issn = {2054-5703}, abstract = {Wolbachia, a widespread bacterium which can influence mosquito-borne pathogen transmission, has recently been detected within Anopheles (An.) species that are malaria vectors in Sub-Saharan Africa. Although studies have reported Wolbachia strains in the An. gambiae complex, apparent low density and prevalence rates require confirmation. In this study, wild Anopheles mosquitoes collected from two regions of Guinea were investigated. In contrast with previous studies, RNA was extracted from adult females (n = 516) to increase the chances for the detection of actively expressed Wolbachia genes, determine Wolbachia prevalence rates and estimate relative strain densities. Molecular confirmation of mosquito species and Wolbachia multilocus sequence typing (MLST) were carried out to analyse phylogenetic relationships of mosquito hosts and newly discovered Wolbachia strains. Strains were detected in An. melas (prevalence rate of 11.6%-16/138) and hybrids between An. melas and An. gambiae sensu stricto (prevalence rate of 40.0%-6/15) from Senguelen in the Maferinyah region. Furthermore, a novel high-density strain, termed wAnsX, was found in an unclassified Anopheles species. The discovery of novel Wolbachia strains (particularly in members, and hybrids, of the An. gambiae complex) provides further candidate strains that could be used for future Wolbachia-based malaria biocontrol strategies.}, } @article {pmid33857432, year = {2021}, author = {Walker, T and Quek, S and Jeffries, CL and Bandibabone, J and Dhokiya, V and Bamou, R and Kristan, M and Messenger, LA and Gidley, A and Hornett, EA and Anderson, ER and Cansado-Utrilla, C and Hegde, S and Bantuzeko, C and Stevenson, JC and Lobo, NF and Wagstaff, SC and Nkondjio, CA and Irish, SR and Heinz, E and Hughes, GL}, title = {Stable high-density and maternally inherited Wolbachia infections in Anopheles moucheti and Anopheles demeilloni mosquitoes.}, journal = {Current biology : CB}, volume = {31}, number = {11}, pages = {2310-2320.e5}, pmid = {33857432}, issn = {1879-0445}, support = {/WT_/Wellcome Trust/United Kingdom ; R21 AI124452/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; BB/T001240/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; V011278/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; *Anopheles/genetics ; In Situ Hybridization, Fluorescence ; *Malaria ; Maternal Inheritance ; Mosquito Vectors ; *Wolbachia/genetics ; }, abstract = {Wolbachia, a widespread bacterium that can reduce pathogen transmission in mosquitoes, has recently been reported to be present in Anopheles (An.) species. In wild populations of the An. gambiae complex, the primary vectors of Plasmodium malaria in Sub-Saharan Africa, Wolbachia DNA sequences at low density and infection frequencies have been detected. As the majority of studies have used highly sensitive nested PCR as the only method of detection, more robust evidence is required to determine whether Wolbachia strains are established as endosymbionts in Anopheles species. Here, we describe high-density Wolbachia infections in geographically diverse populations of An. moucheti and An. demeilloni. Fluorescent in situ hybridization localized a heavy infection in the ovaries of An. moucheti, and maternal transmission was observed. Genome sequencing of both Wolbachia strains obtained genome depths and coverages comparable to those of other known infections. Notably, homologs of cytoplasmic incompatibility factor (cif) genes were present, indicating that these strains possess the capacity to induce the cytoplasmic incompatibility phenotype, which allows Wolbachia to spread through host populations. These strains should be further investigated as candidates for use in Wolbachia biocontrol strategies in Anopheles aiming to reduce the transmission of malaria.}, } @article {pmid33853965, year = {2021}, author = {Edenborough, KM and Flores, HA and Simmons, CP and Fraser, JE}, title = {Using Wolbachia to Eliminate Dengue: Will the Virus Fight Back?.}, journal = {Journal of virology}, volume = {95}, number = {13}, pages = {e0220320}, pmid = {33853965}, issn = {1098-5514}, mesh = {Adaptation, Physiological/*physiology ; Aedes/drug effects/*microbiology ; Animals ; Dengue/pathology/*prevention & control/transmission ; Dengue Virus/*growth & development ; Drosophila/microbiology ; Evolution, Molecular ; Humans ; Insecticide Resistance/physiology ; Mosquito Vectors/microbiology ; Selection, Genetic/genetics ; Wolbachia/*metabolism ; }, abstract = {Recent field trials have demonstrated that dengue incidence can be substantially reduced by introgressing strains of the endosymbiotic bacterium Wolbachia into Aedes aegypti mosquito populations. This strategy relies on Wolbachia reducing the susceptibility of Ae. aegypti to disseminated infection by positive-sense RNA viruses like dengue. However, RNA viruses are well known to adapt to antiviral pressures. Here, we review the viral infection stages where selection for Wolbachia-resistant virus variants could occur. We also consider the genetic constraints imposed on viruses that alternate between vertebrate and invertebrate hosts, and the likely selection pressures to which dengue virus might adapt in order to be effectively transmitted by Ae. aegypti that carry Wolbachia. While there are hurdles to dengue viruses developing resistance to Wolbachia, we suggest that long-term surveillance for resistant viruses should be an integral component of Wolbachia-introgression biocontrol programs.}, } @article {pmid33847843, year = {2021}, author = {Hu, L and Yang, C and Hui, Y and Yu, J}, title = {Mosquito Control Based on Pesticides and Endosymbiotic Bacterium Wolbachia.}, journal = {Bulletin of mathematical biology}, volume = {83}, number = {5}, pages = {58}, pmid = {33847843}, issn = {1522-9602}, mesh = {Animals ; *Culicidae/microbiology ; Humans ; *Models, Biological ; *Mosquito Control/methods/standards ; *Pesticides ; Population Dynamics ; Vector Borne Diseases/prevention & control ; *Wolbachia/physiology ; }, abstract = {Mosquito-borne diseases, such as dengue fever and Zika, have posed a serious threat to human health around the world. Controlling vector mosquitoes is an effective method to prevent these diseases. Spraying pesticides has been the main approach of reducing mosquito population, but it is not a sustainable solution due to the growing insecticide resistance. One promising complementary method is the release of Wolbachia-infected mosquitoes into wild mosquito populations, which has been proven to be a novel and environment-friendly way for mosquito control. In this paper, we incorporate consideration of releasing infected sterile mosquitoes and spraying pesticides to aim to reduce wild mosquito populations based on the population replacement model. We present the estimations for the number of wild mosquitoes or infection density in a normal environment and then discuss how to offset the effect of the heatwave, which can cause infected mosquitoes to lose Wolbachia infection. Finally, we give the waiting time to suppress wild mosquito population to a given threshold size by numerical simulations.}, } @article {pmid33834007, year = {2021}, author = {Wei, Y and Wang, J and Wei, YH and Song, Z and Hu, K and Chen, Y and Zhou, G and Zhong, D and Zheng, X}, title = {Vector Competence for DENV-2 Among Aedes albopictus (Diptera: Culicidae) Populations in China.}, journal = {Frontiers in cellular and infection microbiology}, volume = {11}, number = {}, pages = {649975}, pmid = {33834007}, issn = {2235-2988}, mesh = {*Aedes ; Animals ; China ; *Dengue Virus ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Aedes albopictus is a vector of over 20 arboviruses that has spread throughout the world, mainly in the second half of the twentieth century. Approximately 50-100 million people are infected with dengue virus (DENV) transmitted by Aedes mosquitoes each year, leading to heavy economic burdens for both governments and individuals, among countless other negative consequences. Understanding the vector competence of vector species is critical for effectively preventing and controlling vector-borne diseases. Accordingly, in this study, vector competence was evaluated by quantitative analysis of DENV-2 loads in mosquito tissues (midguts, heads, and salivary glands) and whole mosquitoes through real-time quantitative polymerase chain reaction (RT-qPCR) analysis. Wolbachia and the expression of immune-associated genes (Rel1, Rel2, Dicer2, and STAT) in mosquitoes were also detected by RT-qPCR to explore their impact on vector competence. The amount of DENV-2 in the mosquito midguts, heads, and salivary glands from southern-western China were found to be lower than those from eastern-central-northern China. The DENV-2 loads in whole mosquitoes showed a negative correlation with Rel1 gene (r = -0.285, P = 0.011) and STAT gene expression levels (r = -0.289, P = 0.009). In terms of Wolbachia strains, the density of the wAlbB strain was found to be significantly higher than that of the wAlbA strain in the eight Ae. albopictus populations, and the relative density of the wAlbB strain in mosquitoes from southern-western China was higher than those from eastern-central-northern China. The relative density of the wAlbB strain showed a negative correlation with the mean loads of DENV-2 in the heads (r = -0.729, P = 0.040), salivary glands (r = -0.785, P = 0.021), and whole mosquitoes (r = -0.909, P = 0.002). Thus, there are lower DENV-2 loads in the mosquitoes from southern-western China, which may be related to the innate immunity of mosquitoes as affected by Rel1 in the Toll pathway, STAT in the JAK-STAT pathway, and the relative density of the wAlbB strain.}, } @article {pmid33822117, year = {2021}, author = {Che-Mendoza, A and Martin-Park, A and Chávez-Trava, JM and Contreras-Perera, Y and Delfín-González, H and González-Olvera, G and Leirana-Alcocer, J and Guillermo-May, G and Chan-Espinoza, D and Pavia-Ruz, N and Méndez-Vales, RE and Alcocer-Gamboa, A and Correa-Morales, F and Palacio-Vargas, J and Zhang, D and Vazquez-Prokopec, G and Xi, Z and Manrique-Saide, P}, title = {Abundance and Seasonality of Aedes aegypti (Diptera: Culicidae) in Two Suburban Localities of South Mexico, With Implications for Wolbachia (Rickettsiales: Rickettsiaceae)-Carrying Male Releases for Population Suppression.}, journal = {Journal of medical entomology}, volume = {58}, number = {4}, pages = {1817-1825}, pmid = {33822117}, issn = {1938-2928}, mesh = {*Aedes/microbiology/physiology ; Animals ; Arbovirus Infections/prevention & control/transmission ; Infertility, Male/*microbiology ; Male ; Mexico/epidemiology ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/physiology ; Population Control/methods ; Seasons ; *Wolbachia ; }, abstract = {We conducted a baseline characterization of the abundance and seasonality of Aedes aegypti (Linnaeus, 1762)-a vector of dengue, chikungunya, and Zika-in two suburban localities of Yucatan, Mexico, as the first step in the implementation of an integrated vector management (IVM) plan combining 'traditional Aedes control' (source reduction/truck-mounted ultra-low volume [ULV] spraying) and incompatible insect technique/sterile insect technique for population suppression in Yucatan, Mexico. Weekly entomological collections with ovitraps and BG-sentinel traps were performed in 1-ha quadrants of both localities for 1 yr. Three distinct periods/phases were identified, closely associated with precipitation: 1) a phase of low population abundance during the dry season (weekly average of Aedes eggs per ovitrap and adults per BG trap = 15.51 ± 0.71 and 10.07 ± 0.88, respectively); 2) a phase of population growth and greatest abundance of Aedes (49.03 ± 1.48 eggs and 25.69 ± 1.31 adults) during the rainy season; and finally 3) a phase of decline among populations (20.91 ± 0.97 eggs and 3.24 ± 0.21 adults) after the peak of the rainy season. Seasonal abundance and dynamics of Ae. aegypti populations suggest that it is feasible to develop and implement time-specific actions as part of an IVM approach incorporating integrating novel technologies (such as rear-and-release of Wolbachia-infected males) with classic (insecticide-based) approaches implemented routinely for vector control. In agreement with the local vector control program, we propose a pilot IVM strategy structured in a preparation phase, an attack phase with traditional vector control, and a suppression phase with inundative releases, which are described in this paper.}, } @article {pmid33798671, year = {2021}, author = {Jiang, N and Xue, DY and Han, HX and Cheng, R}, title = {Estimating hybridization as a consequence of climatic fluctuations: A case study of two geometridae species.}, journal = {Molecular phylogenetics and evolution}, volume = {161}, number = {}, pages = {107168}, doi = {10.1016/j.ympev.2021.107168}, pmid = {33798671}, issn = {1095-9513}, mesh = {Animals ; *Climate Change ; DNA, Mitochondrial/genetics ; Female ; Gene Flow ; *Hybridization, Genetic ; Male ; Moths/*genetics ; Phylogeny ; Sequence Analysis, DNA ; }, abstract = {In some cases, the phylogenetic analysis based on the mitochondrial DNA (mtDNA) and the nuclear DNA (ncDNA) are discordant. There are three major causes of the discordance within insects, including hybridization, incomplete lineage sorting (ILS) and infection by Wolbachia. In this study, we used a combination of multilocus and coalescent analyses to explore these processes occurred during the evolutionary history of Limbatochlamys rosthorni Rothschild, 1894 and Limbatochlamys pararosthorni Han and Xue, 2005. The ncDNA phylogenetic tree supported two reciprocally monophyletic species, whereas the mtDNA results failed to reveal such a structure and revealed an extensive level of admixture between two species. Because of very low Wolbachia infection rates (<20%), we firstly excluded this reason for the discordance. The fixed nucleotide differences and large genetic distances (1.5-2.5%) at the ncDNA genes suggested that the lineage sorting process between these two species is nearly complete and two species have experienced a prolonged period of independent evolution. Thus, we secondly excluded ILS. Sharing haplotypes, mtDNA gene flow occurring and the transitional samples with morphological features supported hybridization. The distribution contraction during glaciations and postglacial distribution expansion might have facilitated hybridization. Taken together, our study indicates that the current genetic structure of L. rosthorni and L. pararosthorni is the results of contraction and fragmentation into separated refugia during glaciations, followed by postglacial expansion and admixture.}, } @article {pmid33794350, year = {2021}, author = {Konecka, E and Olszanowski, Z}, title = {Wolbachia supergroup E found in Hypochthonius rufulus (Acari: Oribatida) in Poland.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {91}, number = {}, pages = {104829}, doi = {10.1016/j.meegid.2021.104829}, pmid = {33794350}, issn = {1567-7257}, mesh = {Animals ; Mites/*microbiology ; Phylogeny ; Poland ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Data on the spread of intracellular bacteria in oribatid mites (Acari: Oribatida) are scarce. Our work fills a gap in the research on endosymbionts in this group of invertebrates and provides information on Wolbachia infection in Hypochthonius rufulus (Acari: Oribatida) from soil, litter and moss sample collected in south-eastern Poland. This is the first report of Wolbachia in H. rufulus. Phylogeny based on the analysis of the 16S rRNA, gatB, fbpA, gltA, ftsZ and hcpA gene sequences revealed that Wolbachia from H. rufulus represented supergroup E and was related to bacterial endosymbionts of Collembola. The unique sequence within Wolbachia supergroup E was detected for the 16S rRNA gene of the bacteria from H. rufulus. The sequences of Wolbachia 16S rRNA and housekeeping genes have been deposited in publicly available databases and are an important source of molecular data for comparative studies.}, } @article {pmid33776981, year = {2021}, author = {Flatau, R and Segoli, M and Hawlena, H}, title = {Wolbachia Endosymbionts of Fleas Occur in All Females but Rarely in Males and Do Not Show Evidence of Obligatory Relationships, Fitness Effects, or Sex-Distorting Manipulations.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {649248}, pmid = {33776981}, issn = {1664-302X}, abstract = {The widespread temporal and spatial persistence of endosymbionts in arthropod host populations, despite potential conflicts with their hosts and fluctuating environmental conditions, is puzzling. Here, we disentangled three main mechanisms that are commonly proposed to explain such persistence, namely, obligatory relationships, in which the host is fully dependent on its endosymbiont, fitness advantages conferred by the endosymbiont, and reproductive manipulations imposed by the endosymbiont. Our model system reflects an extreme case, in which the Wolbachia endosymbiont persists in all female flea hosts but rarely in male ones. We cured fleas of both sexes of Wolbachia but found no indications for either lower reproduction, offspring survival, or a change in the offspring sex ratio, compared to Wolbacia-infected fleas. These results do not support any of the suggested mechanisms. We highlight future directions to advance our understanding of endosymbiont persistence in fleas, as well as in other model systems, with extreme sex-differences in endosymbiont persistence. Insights from such studies are predicted to shed light on the evolution and ecology of arthropod-endosymbiont interactions in nature.}, } @article {pmid33764469, year = {2021}, author = {Pilgrim, J and Thongprem, P and Davison, HR and Siozios, S and Baylis, M and Zakharov, EV and Ratnasingham, S and deWaard, JR and Macadam, CR and Smith, MA and Hurst, GDD}, title = {Torix Rickettsia are widespread in arthropods and reflect a neglected symbiosis.}, journal = {GigaScience}, volume = {10}, number = {3}, pages = {}, pmid = {33764469}, issn = {2047-217X}, support = {BB/M011186/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; *Arthropods/genetics ; Base Sequence ; Humans ; Phylogeny ; *Rickettsia/genetics ; Symbiosis ; }, abstract = {BACKGROUND: Rickettsia are intracellular bacteria best known as the causative agents of human and animal diseases. Although these medically important Rickettsia are often transmitted via haematophagous arthropods, other Rickettsia, such as those in the Torix group, appear to reside exclusively in invertebrates and protists with no secondary vertebrate host. Importantly, little is known about the diversity or host range of Torix group Rickettsia.

RESULTS: This study describes the serendipitous discovery of Rickettsia amplicons in the Barcode of Life Data System (BOLD), a sequence database specifically designed for the curation of mitochondrial DNA barcodes. Of 184,585 barcode sequences analysed, Rickettsia is observed in ∼0.41% of barcode submissions and is more likely to be found than Wolbachia (0.17%). The Torix group of Rickettsia are shown to account for 95% of all unintended amplifications from the genus. A further targeted PCR screen of 1,612 individuals from 169 terrestrial and aquatic invertebrate species identified mostly Torix strains and supports the "aquatic hot spot" hypothesis for Torix infection. Furthermore, the analysis of 1,341 SRA deposits indicates that Torix infections represent a significant proportion of all Rickettsia symbioses found in arthropod genome projects.

CONCLUSIONS: This study supports a previous hypothesis that suggests that Torix Rickettsia are overrepresented in aquatic insects. In addition, multiple methods reveal further putative hot spots of Torix Rickettsia infection, including in phloem-feeding bugs, parasitoid wasps, spiders, and vectors of disease. The unknown host effects and transmission strategies of these endosymbionts make these newly discovered associations important to inform future directions of investigation involving the understudied Torix Rickettsia.}, } @article {pmid33760987, year = {2021}, author = {Abrun, P and Ashouri, A and Duplouy, A and Farahani, HK}, title = {Wolbachia impairs post-eclosion host preference in a parasitoid wasp.}, journal = {Die Naturwissenschaften}, volume = {108}, number = {2}, pages = {13}, pmid = {33760987}, issn = {1432-1904}, support = {120586//Marie-Curie Sklodowska Individual fellowship/ ; }, mesh = {Animals ; Female ; Host-Parasite Interactions/*physiology ; Moths/*parasitology ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {Host preference behavior can result in adaptive advantages with important consequences for the fitness of individuals. Hopkin's host-selection principle (HHSP) suggests that organisms at higher trophic levels demonstrate a preference for the host species on which they developed during their own larval stage. Although investigated in many herbivorous and predatory insects, the HHSP has, to our knowledge, never been tested in the context of insects hosting selfish endosymbiotic passengers. Here, we investigated the effect of infection with the facultative bacterial symbiont Wolbachia on post-eclosion host preference in the parasitoid wasp Trichogramma brassicae (Hymenoptera: Trichogrammatidae). We compared host preference in Wolbachia-infected individuals and uninfected adult female parasitoids after rearing them on two different Lepidopteran hosts, namely the flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) or the grain moth Sitotroga cerealella (Lepidoptera: Gelechiidae) in choice and no choice experimental design (n = 120 wasps per each choice/no choice experiments). We showed that in T. brassicae, Wolbachia affects the post-eclosion host preference of female wasps. Wolbachia-infected wasps did not show any host preference and more frequently switched hosts in the laboratory, while uninfected wasps significantly preferred to lay eggs on the host species they developed on. Additionally, Wolbachia significantly improved the emergence rate of infected wasps when reared on new hosts. Altogether, our results revealed that the wasp's infection with Wolbachia may lead to impairment of post-eclosion host preference and facilitates growing up on different host species. The impairment of host preference by Wolbachia may allow T. brassicae to shift between hosts, a behavior that might have important evolutionary consequences for the wasp and its symbiont.}, } @article {pmid33717014, year = {2021}, author = {Wang, YT and Shen, RX and Xing, D and Zhao, CP and Gao, HT and Wu, JH and Zhang, N and Zhang, HD and Chen, Y and Zhao, TY and Li, CX}, title = {Metagenome Sequencing Reveals the Midgut Microbiota Makeup of Culex pipiens quinquefasciatus and Its Possible Relationship With Insecticide Resistance.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {625539}, pmid = {33717014}, issn = {1664-302X}, abstract = {Midgut microbiota can participate in the detoxification and metabolism processes in insects, but there are few reports on the relationship between midgut microbiota and insecticide resistance in mosquitoes. In this study, we performed metagenomic sequencing on a susceptible strain (SS), a field-collected Hainan strain (HN), and a deltamethrin-resistant strain (RR) of Culex pipiens quinquefasciatus to understand the diversity and functions of their midgut microbiota. The results revealed differences in midgut microbiota among the three strains of Cx. pipiens quinquefasciatus. At the phylum level, Proteobacteria was the most prominent, accounting for nearly 70% of their midgut microbes. At the genus level, Aeromonas made up the highest proportion. In addition, Aeromonas, Morganella, Elizabethkingia, Enterobacter, Cedecea, and Thorsellia showed significant differences between strains. At the species level, Bacillus cereus, Enterobacter cloacae complex sp. 4DZ3-17B2, Streptomyces sp. CNQ329, and some species of Pseudomonas and Wolbachia were more abundant in the two resistant strains. Principal component analysis (PCA) showed that the SS strain had significantly different metagenomic functions than the two deltamethrin-resistant strains (HN and RR strain). The HN and RR strains differed from the SS strain in more than 10 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The analysis of species abundance and functional diversity can provide directions for future studies.}, } @article {pmid33716790, year = {2021}, author = {Martins, M and Ramos, LFC and Murillo, JR and Torres, A and de Carvalho, SS and Domont, GB and de Oliveira, DMP and Mesquita, RD and Nogueira, FCS and Maciel-de-Freitas, R and Junqueira, M}, title = {Comprehensive Quantitative Proteome Analysis of Aedes aegypti Identifies Proteins and Pathways Involved in Wolbachia pipientis and Zika Virus Interference Phenomenon.}, journal = {Frontiers in physiology}, volume = {12}, number = {}, pages = {642237}, pmid = {33716790}, issn = {1664-042X}, abstract = {Zika virus (ZIKV) is a global public health emergency due to its association with microcephaly, Guillain-Barré syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of ZIKV, distributed across four continents, and no antivirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboviruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry (MS)-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the A. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in reactive oxygen species production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by MS and corroborates the idea that Wolbachia helps to block ZIKV infections in A. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in A. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.}, } @article {pmid33704487, year = {2021}, author = {Dobson, SL}, title = {When More is Less: Mosquito Population Suppression Using Sterile, Incompatible and Genetically Modified Male Mosquitoes.}, journal = {Journal of medical entomology}, volume = {58}, number = {5}, pages = {1980-1986}, doi = {10.1093/jme/tjab025}, pmid = {33704487}, issn = {1938-2928}, mesh = {Animals ; *Culicidae/microbiology/radiation effects ; Male ; Mosquito Control/*methods ; *Mosquito Vectors/microbiology/radiation effects ; Wolbachia/*physiology ; }, abstract = {The current review of the Sterile Insect Technique (SIT) is motivated by new technologies and the recent renaissance of male release field trials, which is driving an evolution in mosquito control and regulation. Practitioners that are releasing male mosquitoes would do well to learn from past successes and failures, including political and public engagement complications. With examples that include nuanced integrations of the different technologies, e.g., combinations of Wolbachia and irradiation, it is critical that scientists understand and communicate accurately about the technologies, including their evolving management by different regulatory agencies in the USA. Some male release approaches are considered 'pesticides' and regulated by federal and state agencies, while other male release approaches are unregulated. It is important to consider how the new technologies fit with the more 'traditional' chemical applications of adulticides and larvicides. The economics of male release programs are substantially different from traditional control costs, which can be a challenge to their adoption by abatement districts. However, there is substantial need to overcome these complications and challenges, because the problem with invasive mosquitoes grows ever worse with factors that include insecticide resistance, globalization and climate change.}, } @article {pmid33683351, year = {2021}, author = {Shropshire, JD and Rosenberg, R and Bordenstein, SR}, title = {The impacts of cytoplasmic incompatibility factor (cifA and cifB) genetic variation on phenotypes.}, journal = {Genetics}, volume = {217}, number = {1}, pages = {1-13}, pmid = {33683351}, issn = {1943-2631}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster ; Female ; Genetic Fitness ; *Genetic Variation ; Infertility/microbiology ; Male ; *Phenotype ; Prophages/*genetics ; Spermatozoa/microbiology ; Viral Proteins/*genetics ; Wolbachia/pathogenicity/virology ; }, abstract = {Wolbachia are maternally transmitted, intracellular bacteria that can often selfishly spread through arthropod populations via cytoplasmic incompatibility (CI). CI manifests as embryonic death when males expressing prophage WO genes cifA and cifB mate with uninfected females or females harboring an incompatible Wolbachia strain. Females with a compatible cifA-expressing strain rescue CI. Thus, cif-mediated CI confers a relative fitness advantage to females transmitting Wolbachia. However, whether cif sequence variation underpins incompatibilities between Wolbachia strains and variation in CI penetrance remains unknown. Here, we engineer Drosophila melanogaster to transgenically express cognate and non-cognate cif homologs and assess their CI and rescue capability. Cognate expression revealed that cifA;B native to D. melanogaster causes strong CI, and cognate cifA;B homologs from two other Drosophila-associated Wolbachia cause weak transgenic CI, including the first demonstration of phylogenetic type 2 cifA;B CI. Intriguingly, non-cognate expression of cifA and cifB alleles from different strains revealed that cifA homologs generally contribute to strong transgenic CI and interchangeable rescue despite their evolutionary divergence, and cifB genetic divergence contributes to weak or no transgenic CI. Finally, we find that a type 1 cifA can rescue CI caused by a genetically divergent type 2 cifA;B in a manner consistent with unidirectional incompatibility. By genetically dissecting individual CI functions for type 1 and 2 cifA and cifB, this work illuminates new relationships between cif genotype and CI phenotype. We discuss the relevance of these findings to CI's genetic basis, phenotypic variation patterns, and mechanism.}, } @article {pmid33671250, year = {2021}, author = {Zheng, Z and Hu, X and Xu, Y and Wei, C and He, H}, title = {Bacterial Composition and Diversity of the Digestive Tract of Odontomachus monticola Emery and Ectomomyrmex javanus Mayr.}, journal = {Insects}, volume = {12}, number = {2}, pages = {}, pmid = {33671250}, issn = {2075-4450}, support = {Grant Number 31570388//the Natural Science Foundation of China/ ; Grant Number 2452019174.//the Fundamental Research Funds for the Central Universities/ ; }, abstract = {Ponerine ants are generalist predators feeding on a variety of small arthropods, annelids, and isopods; however, knowledge of their bacterial communities is rather limited. This study investigated the bacterial composition and diversity in the digestive tract (different gut sections and the infrabuccal pockets (IBPs)) of two ponerine ant species (Odontomachus monticola Emery and Ectomomyrmex javanus Mayr) distributed in northwestern China using high-throughput sequencing. We found that several dominant bacteria that exist in other predatory ants were also detected in these two ponerine ant species, including Wolbachia, Mesoplasma, and Spiroplasma. Bacterial communities of these two ant species were differed significantly from each other, and significant differences were also observed across their colonies, showing distinctive inter-colony characteristics. Moreover, bacterial communities between the gut sections (crops, midguts, and hindguts) of workers were highly similar within colony, but they were clearly different from those in IBPs. Further, bacterial communities in the larvae of O. monticola were similar to those in the IBPs of workers, but significantly different from those in gut sections. We presume that the bacterial composition and diversity in ponerine ants are related to their social behavior and feeding habits, and bacterial communities in the IBPs may play a potential role in their social life.}, } @article {pmid33664278, year = {2021}, author = {Ma, YJ and He, HP and Zhao, HM and Xian, YD and Guo, H and Liu, B and Xue, K}, title = {Microbiome diversity of cotton aphids (Aphis gossypii) is associated with host alternation.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {5260}, pmid = {33664278}, issn = {2045-2322}, mesh = {Animals ; Aphids/genetics/*microbiology ; Genetic Variation/*genetics ; Gossypium/genetics/microbiology ; High-Throughput Nucleotide Sequencing ; Host-Pathogen Interactions/*genetics ; Microbiota/*genetics ; Symbiosis/genetics ; }, abstract = {Aphids are infected by a series of bacteria that can help them survive on specific host plants. However, the associations between aphids and these bacteria are not clear, and the bacterial communities in many aphid species are poorly characterized. Here, we investigated the bacterial communities of cotton aphids (Aphis gossypii) on 2 representative winter host plants and transferred to 3 summer host plants by 16S rDNA sequencing using the Illumina MiSeq platform. Our results revealed that the bacterial communities varied among cotton aphids on hibiscus, cotton aphids on pomegranate, cotton aphids on cotton transferred from hibiscus, cotton aphids on muskmelon transferred from hibiscus, cotton aphids on cucumber transferred from hibiscus,. The diversity and richness of the bacterial communities were significantly higher in aphids on muskmelon and aphids on cucumber than in the other treatments. There were two main factors influencing the distribution of internal bacterial OTUs revealed by principal component analysis, including the differences among Punicaceae, Malvaceae and Cucurbitaceae. There were 28 bacterial communities with significant differences between two arbitrary treatments, which could be grouped into 6 main clusters depending on relative abundance. Moreover, our results indicated that in addition to the obligate endosymbiont Buchnera, with a dominant position (> 52%), A. gossypii also harbored 3 facultative endosymbiotic bacteria (Serratia, Arsenophonus, and Wolbachia) and 3 possibly symbiotic bacteria (Acinetobacter, Pantoea, and Flavobacterium). There were several correspondences between the symbiotic bacteria in cotton aphids and the specific host plants of the aphids. This study provides a better understanding of the interactions among symbiotic bacteria, aphids and host plants, suggesting that the selection pressure on aphid bacterial communities is likely to be exerted by the species of host plants.}, } @article {pmid33659853, year = {2020}, author = {Romanov, DA and Zakharov, IA and Shaikevich, EV}, title = {Wolbachia, Spiroplasma, and Rickettsia symbiotic bacteria in aphids (Aphidoidea).}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {24}, number = {6}, pages = {673-682}, doi = {10.18699/VJ20.661}, pmid = {33659853}, issn = {2500-0462}, abstract = {Aphids are a diverse family of crop pests. Aphids formed a complex relationship with intracellular bacteria. Depending on the region of study, the species composition of both aphids and their facultative endosymbionts varies. The aim of the work was to determine the occurrence and genetic diversity of Wolbachia, Spiroplasma and Rickettsia symbionts in aphids collected in 2018-2019 in Moscow. For these purposes, 578 aphids from 32 collection sites were tested by PCR using specific primers. At least 21 species of aphids from 14 genera and four families were identified by barcoding method, of which 11 species were infected with endosymbionts. Rickettsia was found in six species, Wolbachia in two species, Spiroplasma in one species. The presence of Rickettsia in Impatientinum asiaticum, Myzus cerasi, Hyalopterus pruni, Eucallipterus tiliae, Chaitophorus tremulae and Wolbachia in Aphis pomi and C. tremulae has been described for the first time. A double infection with Rickettsia and Spiroplasma was detected in a half of pea aphid (Acyrthosiphon pisum) individuals. For the first time was found that six species of aphids are infected with Rickettsia that are genetically different from previously known. It was first discovered that A. pomi is infected with two Wolbachia strains, one of which belongs to supergroup B and is genetically close to Wolbachia from C. tremulae. The second Wolbachia strain from A. pomi belongs to the supergroup M, recently described in aphid species. Spiroplasma, which we observed in A. pisum, is genetically close to male killing Spiroplasma from aphids, ladybirds and moths. Both maternal inheritance and horizontal transmission are the pathways for the distribution of facultative endosymbiotic bacteria in aphids.}, } @article {pmid33646378, year = {2021}, author = {Wangkeeree, J and Sanit, P and Roddee, J and Hanboonsong, Y}, title = {Phylogeny and Strain Typing of Wolbachia from Yamatotettix flavovittatus Matsumura Leafhoppers.}, journal = {Current microbiology}, volume = {78}, number = {4}, pages = {1367-1376}, pmid = {33646378}, issn = {1432-0991}, support = {226/2563//National Research Council of Thailand/ ; }, mesh = {Animals ; Female ; *Hemiptera ; Multilocus Sequence Typing ; Phylogeny ; *Saccharum ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a maternally inherited bacterium of insects that can alter the reproduction, biology, and fitness of the hosts. It was detected in natural populations of the Yamatotettix flavovittatus Matsumura leafhoppers, the vector of phytoplasma, which is responsible for sugarcane white leaf disease. Wolbachia infection prolongs the longevity of female leafhoppers and promotes a strong reproductive incompatibility; importantly, highly maternal transmission rate was observed. However, limited data on the diversity or strain typing of Wolbachia in Y. flavovittatus are available. We aimed here to detect the presence of Wolbachia in different populations by amplification of the wsp gene, which was then sequenced. Multilocus sequence typing (MLST) was also performed to explore the diversity of the Wolbachia strains. Based on the wsp sequences, Wolbachia in the Y. flavovittatus leafhoppers belonged to supergroup B, and formed a distinct evolutionary lineage; therefore, we designated this new specific strain as wYfla. The MLST profiles revealed ten potential new sequence types (STs) in different leafhopper populations. Multiple STs were detected in individual leafhoppers, among which the ST-wYfla1 strain was predominant. Furthermore, we obtained congruent results for the phylogenetic analyses using the wsp gene and MLST loci. To the best of our knowledge, this is the first study characterizing Wolbachia strains in Y. flavovittatus. Our results reveal a novel strain and multiple STs of Wolbachia, and these data may prove useful in the exploitation of Wolbachia as a biological Y. flavovittatus control agent.}, } @article {pmid33644932, year = {2021}, author = {Burdina, EV and Bykov, RA and Menshanov, PN and Ilinsky, YY and Gruntenko, NЕ}, title = {Unique Wolbachia strain wMelPlus increases heat stress resistance in Drosophila melanogaster.}, journal = {Archives of insect biochemistry and physiology}, volume = {106}, number = {4}, pages = {e21776}, doi = {10.1002/arch.21776}, pmid = {33644932}, issn = {1520-6327}, support = {0259-2021-0016//Ministry of Science and Higher Education of the Russian Federation/ ; 20-04-00579//Russian Foundation for Basic Research/ ; }, mesh = {Alkaline Phosphatase/metabolism ; Animals ; Dopamine/metabolism ; Drosophila melanogaster/metabolism/*microbiology ; Genotype ; Heat-Shock Response ; Symbiosis/physiology ; *Thermotolerance/genetics/physiology ; Wolbachia/*genetics ; }, abstract = {Maternally inherited endosymbiotic bacterium Wolbachia infects Drosophila melanogaster populations worldwide. Its genetic diversity includes several closely related genotypes, which can be attributed to two main genotype groups: wMel and wMelCS. Here, we studied eight D. melanogaster lines carrying the nuclear background of wild type interbred Bi90 line and cytoplasmic backgrounds with or without Wolbachia of different origin, each of which belongs to wMelCS genotype group. We analyzed the effect these seven Wolbachia strains had on the heat stress resistance and dopamine metabolism in D. melanogaster females. Survival under heat stress (38°C, 3 h 30 min) was increased only in the line infected with bacteria of the wMelPlus strain. At the same time, the activity of alkaline phosphatase (an enzyme regulating the pool of dopamine precursor tyrosine) was increased under normal conditions in females infected with all strains under study and retained the response to heat stress typical for the uninfected line. Thus, we found the unique Wolbachia strain that provides an increase of the host stress resistance, and demonstrated that the mechanism of this resistance is not dopamine-mediated.}, } @article {pmid33629719, year = {2021}, author = {Benedict, MQ}, title = {Sterile Insect Technique: Lessons From the Past.}, journal = {Journal of medical entomology}, volume = {58}, number = {5}, pages = {1974-1979}, doi = {10.1093/jme/tjab024}, pmid = {33629719}, issn = {1938-2928}, mesh = {Mosquito Control/instrumentation/*methods/statistics & numerical data ; *Mosquito Vectors ; }, abstract = {When E.F. Knipling conceived of the release of sexually sterile insects to suppress wild populations, he laid down several fundamental qualities that characterized suitable target species-some of which mosquitoes generally violate-including high reproductive rates and large population numbers. Regardless of this, their global importance in public health has led numerous research teams to attempt to use the mosquito sterile insect technique against several species. Because of the degree of financial commitment required for suppression programs, most releases have consisted of preliminary investigations of male performance, population characteristics, and production methods. Those that have accomplished suppression provide important insights regarding the challenges of production, dispersal, and immigration. Insights gained from these studies remain relevant today, regardless of the genetic control technology being applied. In this article, I highlight studies that were notable for the insights that were gained, the intrinsic difficulties that mosquitoes present, and synthesize these into recommendations for successful applications of the sterile insect technique and newer technologies to mosquitoes.}, } @article {pmid33627171, year = {2021}, author = {Bulman, CA and Chappell, L and Gunderson, E and Vogel, I and Beerntsen, B and Slatko, BE and Sullivan, W and Sakanari, JA}, title = {The Eagle effect in the Wolbachia-worm symbiosis.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {118}, pmid = {33627171}, issn = {1756-3305}, support = {OPP1017584//Bill and Melinda Gates Foundation/ ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Brugia malayi/drug effects/microbiology/*physiology ; Doxycycline/pharmacology ; Female ; Male ; Microfilariae/drug effects/*microbiology/physiology ; Onchocerca/drug effects/microbiology/*physiology ; *Symbiosis/drug effects ; Wolbachia/drug effects/*physiology ; }, abstract = {BACKGROUND: Onchocerciasis (river blindness) and lymphatic filariasis (elephantiasis) are two human neglected tropical diseases that cause major disabilities. Mass administration of drugs targeting the microfilarial stage has reduced transmission and eliminated these diseases in several countries but a macrofilaricidal drug that kills or sterilizes the adult worms is critically needed to eradicate the diseases. The causative agents of onchocerciasis and lymphatic filariasis are filarial worms that harbor the endosymbiotic bacterium Wolbachia. Because filarial worms depend on Wolbachia for reproduction and survival, drugs targeting Wolbachia hold great promise as a means to eliminate these diseases.

METHODS: To better understand the relationship between Wolbachia and its worm host, adult Brugia pahangi were exposed to varying concentrations of doxycycline, minocycline, tetracycline and rifampicin in vitro and assessed for Wolbachia numbers and worm motility. Worm motility was monitored using the Worminator system, and Wolbachia titers were assessed by qPCR of the single copy gene wsp from Wolbachia and gst from Brugia to calculate IC50s and in time course experiments. Confocal microscopy was also used to quantify Wolbachia located at the distal tip region of worm ovaries to assess the effects of antibiotic treatment in this region of the worm where Wolbachia are transmitted vertically to the microfilarial stage.

RESULTS: Worms treated with higher concentrations of antibiotics had higher Wolbachia titers, i.e. as antibiotic concentrations increased there was a corresponding increase in Wolbachia titers. As the concentration of antibiotic increased, worms stopped moving and never recovered despite maintaining Wolbachia titers comparable to controls. Thus, worms were rendered moribund by the higher concentrations of antibiotics but Wolbachia persisted suggesting that these antibiotics may act directly on the worms at high concentration. Surprisingly, in contrast to these results, antibiotics given at low concentrations reduced Wolbachia titers.

CONCLUSION: Wolbachia in B. pahangi display a counterintuitive dose response known as the "Eagle effect." This effect in Wolbachia suggests a common underlying mechanism that allows diverse bacterial and fungal species to persist despite exposure to high concentrations of antimicrobial compounds. To our knowledge this is the first report of this phenomenon occurring in an intracellular endosymbiont, Wolbachia, in its filarial host.}, } @article {pmid33610188, year = {2021}, author = {Guo, H and Wang, N and Niu, H and Zhao, D and Zhang, Z}, title = {Interaction of Arsenophonus with Wolbachia in Nilaparvata lugens.}, journal = {BMC ecology and evolution}, volume = {21}, number = {1}, pages = {31}, pmid = {33610188}, issn = {2730-7182}, mesh = {Animals ; China/epidemiology ; *Hemiptera ; Humans ; *Spiroplasma ; Symbiosis ; *Wolbachia ; }, abstract = {BACKGROUND: Co-infection of endosymbionts in the same host is ubiquitous, and the interactions of the most common symbiont Wolbachia with other symbionts, including Spiroplasma, in invertebrate organisms have received increasing attention. However, the interactions between Wolbachia and Arsenophonus, another widely distributed symbiont in nature, are poorly understood. We tested the co-infection of Wolbachia and Arsenophonus in different populations of Nilaparvata lugens and investigated whether co-infection affected the population size of the symbionts in their host.

RESULTS: A significant difference was observed in the co-infection incidence of Wolbachia and Arsenophonus among 5 populations of N. lugens from China, with nearly half of the individuals in the Zhenjiang population harbouring the two symbionts simultaneously, and the rate of occurrence was significantly higher than that of the other 4 populations. The Arsenophonus density in the superinfection line was significantly higher only in the Maanshan population compared with that of the single-infection line. Differences in the density of Wolbachia and Arsenophonus were found in all the tested double-infection lines, and the dominant symbiont species varied with the population only in the Nanjing population, with Arsenophonus the overall dominant symbiont.

CONCLUSIONS: Wolbachia and Arsenophonus could coexist in N. lugens, and the co-infection incidence varied with the geographic populations. Antagonistic interactions were not observed between Arsenophonus and Wolbachia, and the latter was the dominant symbiont in most populations.}, } @article {pmid33103066, year = {2020}, author = {Costa, GB and Smithyman, R and O'Neill, SL and Moreira, LA}, title = {How to engage communities on a large scale? Lessons from World Mosquito Program in Rio de Janeiro, Brazil.}, journal = {Gates open research}, volume = {4}, number = {}, pages = {109}, pmid = {33103066}, issn = {2572-4754}, abstract = {One of the pillars of the World Health Organization's (WHO) Global Vector Control Response 2017-2030 strategy is the engagement of communities. Among the priority activities, defined by 2022 by the WHO, is the development of plans for the effective engagement and mobilisation of communities in vector control. Novel technologies for arboviruses control are being developed, such as the Wolbachia method, implemented by the World Mosquito Program (WMP). Here we discuss and analyse the framework for community engagement implemented by the WMP in Brazil, during the large-scale deployment of the method in the municipalities of Niterói and Rio de Janeiro, Brazil. Our experience indicates that the community engagement work for arboviruses control should be understood as an opportunity for local development. It is necessary, based on an integrated analysis of the territory, to understand that the actions for arboviruses control could be a catalyst for the necessary socioenvironmental, cultural and public health changes. Furthermore, it is essential to understand that community engagement goes beyond informing or asking for population consent, but it constitutes a possibility for dialogue and exchange between the various stakeholders present in the territories, to build on cooperation for mosquito-borne disease control.}, } @article {pmid33591971, year = {2021}, author = {Lau, MJ and Ross, PA and Hoffmann, AA}, title = {Infertility and fecundity loss of Wolbachia-infected Aedes aegypti hatched from quiescent eggs is expected to alter invasion dynamics.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {2}, pages = {e0009179}, pmid = {33591971}, issn = {1935-2735}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/physiology ; Animal Husbandry/methods ; Animals ; Female ; Fertility ; Male ; Ovum/physiology ; Pest Control, Biological/*methods ; Temperature ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia shows viral blocking in its mosquito host, leading to its use in arboviral disease control. Releases with Wolbachia strains wMel and wAlbB infecting Aedes aegypti have taken place in several countries. Mosquito egg survival is a key factor influencing population persistence and this trait is also important when eggs are stored prior to releases. We therefore tested the viability of mosquitoes derived from Wolbachia wMel and wAlbB-infected as well as uninfected eggs after long-term storage under diurnal temperature cycles of 11-19°C and 22-30°C. Eggs stored at 11-19°C had higher hatch proportions than those stored at 22-30°C. Adult Wolbachia density declined when they emerged from eggs stored for longer, which was associated with incomplete cytoplasmic incompatibility (CI) when wMel-infected males were crossed with uninfected females. Females from stored eggs at both temperatures continued to show perfect maternal transmission of Wolbachia, but storage reduced the fecundity of both wMel and wAlbB-infected females relative to uninfected mosquitoes. Furthermore, we found a very strong negative impact of the wAlbB infection on the fertility of females stored at 22-30°C, with almost 80% of females hatching after 11 weeks of storage being infertile. Our findings provide guidance for storing Wolbachia-infected A. aegypti eggs to ensure high fitness adult mosquitoes for release. Importantly, they also highlight the likely impact of egg quiescence on the population dynamics of Wolbachia-infected populations in the field, and the potential for Wolbachia to suppress mosquito populations through cumulative fitness costs across warm and dry periods, with expected effects on dengue transmission.}, } @article {pmid33588918, year = {2021}, author = {Farnesi, LC and Carvalho, FD and Lacerda, APC and Moreira, LA and Bruno, RV}, title = {Correction to: The influence of different sources of blood meals on the physiology of Aedes aegypti harboring Wolbachia wMel: mouse blood as an alternative for mosquito rearing.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {109}, pmid = {33588918}, issn = {1756-3305}, } @article {pmid33584729, year = {2020}, author = {Pimentel, AC and Cesar, CS and Martins, M and Cogni, R}, title = {The Antiviral Effects of the Symbiont Bacteria Wolbachia in Insects.}, journal = {Frontiers in immunology}, volume = {11}, number = {}, pages = {626329}, pmid = {33584729}, issn = {1664-3224}, mesh = {Animals ; *Drosophila/immunology/microbiology/virology ; RNA Virus Infections/*immunology ; RNA Viruses/*immunology ; Symbiosis/*immunology ; Wolbachia/*immunology ; }, abstract = {Wolbachia is a maternally transmitted bacterium that lives inside arthropod cells. Historically, it was viewed primarily as a parasite that manipulates host reproduction, but more recently it was discovered that Wolbachia can also protect Drosophila species against infection by RNA viruses. Combined with Wolbachia's ability to invade insect populations due to reproductive manipulations, this provides a way to modify mosquito populations to prevent them transmitting viruses like dengue. In this review, we discuss the main advances in the field since Wolbachia's antiviral effect was discovered 12 years ago, identifying current research gaps and potential future developments. We discuss that the antiviral effect works against a broad range of RNA viruses and depends on the Wolbachia lineage. We describe what is known about the mechanisms behind viral protection, and that recent studies suggest two possible mechanisms: activation of host immunity or competition with virus for cellular resources. We also discuss how association with Wolbachia may influence the evolution of virus defense on the insect host genome. Finally, we investigate whether the antiviral effect occurs in wild insect populations and its ecological relevance as a major antiviral component in insects.}, } @article {pmid33584626, year = {2021}, author = {Scolari, F and Sandionigi, A and Carlassara, M and Bruno, A and Casiraghi, M and Bonizzoni, M}, title = {Exploring Changes in the Microbiota of Aedes albopictus: Comparison Among Breeding Site Water, Larvae, and Adults.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {624170}, pmid = {33584626}, issn = {1664-302X}, abstract = {The mosquito body hosts highly diverse microbes, which influence different physiological traits of both larvae and adults. The composition of adult mosquito microbiota is tightly linked to that of larvae, which are aquatic and feed on organic detritus, algae and prokaryotic microorganisms present in their breeding sites. Unraveling the ecological features of larval habitats that shape the structure of bacterial communities and their interactions with the mosquito host is still a poorly investigated topic in the Asian tiger mosquito Aedes albopictus, a highly invasive species that is vector of numerous arboviruses, including Dengue, Chikungunya, and Zika viruses. In this study, we investigated the composition of the bacterial community present in the water from a natural larval breeding site in which we separately reared wild-collected larvae and hatched eggs of the Foshan reference laboratory strain. Using sequence analysis of bacterial 16S rRNA gene amplicons, we comparatively analyzed the microbiota of the larvae and that of adult mosquitoes, deriving information about the relative impact of the breeding site water on shaping mosquito microbiota. We observed a higher bacterial diversity in breeding site water than in larvae or adults, irrespective of the origin of the sample. Moreover, larvae displayed a significantly different and most diversified microbial community than newly emerged adults, which appeared to be dominated by Proteobacteria. The microbiota of breeding site water significantly increased its diversity over time, suggesting the presence of a dynamic interaction among bacterial communities, breeding sites and mosquito hosts. The analysis of Wolbachia prevalence in adults from Foshan and five additional strains with different geographic origins confirmed the described pattern of dual wAlbA and wAlbB strain infection. However, differences in Wolbachia prevalence were detected, with one strain from La Reunion Island showing up to 18% uninfected individuals. These findings contribute in further understanding the dynamic interactions between the ecology of larval habitats and the structure of host microbiota, as well as providing additional information relative to the patterns of Wolbachia infection.}, } @article {pmid33581654, year = {2021}, author = {Kwarteng, A and Sylverken, A and Asiedu, E and Ahuno, ST}, title = {Genome editing as control tool for filarial infections.}, journal = {Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie}, volume = {137}, number = {}, pages = {111292}, doi = {10.1016/j.biopha.2021.111292}, pmid = {33581654}, issn = {1950-6007}, mesh = {Animals ; CRISPR-Associated Protein 9/genetics/metabolism ; *CRISPR-Cas Systems ; Clustered Regularly Interspaced Short Palindromic Repeats ; Filariasis/genetics/parasitology/*therapy ; Filaricides/therapeutic use ; Filarioidea/drug effects/*genetics/pathogenicity ; *Gene Editing ; *Genetic Therapy ; Humans ; Protozoan Vaccines/therapeutic use ; }, abstract = {Human filarial infections are vector-borne nematode infections, which include lymphatic filariasis, onchocerciasis, loiasis, and mansonella filariasis. With a high prevalence in developing countries, filarial infections are responsible for some of the most debilitating morbidities and a vicious cycle of poverty and disease. Global initiatives set to eradicate these infections include community mass treatments, vector control, provision of care for morbidity, and search for vaccines. However, there are growing challenges associated with mass treatments, vector control, and antifilarial vaccine development. With the emergence of genome editing tools and successful applications in other infectious diseases, the integration of genetic editing techniques in future control strategies for filarial infections would offer the best option for eliminating filarial infections. In this review, we briefly discuss the mechanisms of the three main genetic editing techniques and explore the potential applications of these powerful tools to control filarial infections.}, } @article {pmid33580178, year = {2021}, author = {Kapantaidaki, DE and Antonatos, S and Evangelou, V and Papachristos, DP and Milonas, P}, title = {Genetic and endosymbiotic diversity of Greek populations of Philaenus spumarius, Philaenus signatus and Neophilaenus campestris, vectors of Xylella fastidiosa.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {3752}, pmid = {33580178}, issn = {2045-2322}, mesh = {Animals ; Genetic Variation/genetics ; Greece ; Hemiptera/*genetics/*microbiology ; Insect Vectors/genetics/microbiology ; Olea/microbiology ; Phylogeny ; Plant Diseases/microbiology ; Symbiosis/genetics ; Xylella/*genetics/metabolism/pathogenicity ; }, abstract = {The plant-pathogenic bacterium Xylella fastidiosa which causes significant diseases to various plant species worldwide, is exclusively transmitted by xylem sap-feeding insects. Given the fact that X. fastidiosa poses a serious potential threat for olive cultivation in Greece, the main aim of this study was to investigate the genetic variation of Greek populations of three spittlebug species (Philaenus spumarius, P. signatus and Neophilaenus campestris), by examining the molecular markers Cytochrome Oxidase I, cytochrome b and Internal Transcribed Spacer. Moreover, the infection status of the secondary endosymbionts Wolbachia, Arsenophonus, Hamiltonella, Cardinium and Rickettsia, among these populations, was determined. According to the results, the ITS2 region was the less polymorphic, while the analyzed fragments of COI and cytb genes, displayed high genetic diversity. The phylogenetic analysis placed the Greek populations of P. spumarius into the previously obtained Southwest clade in Europe. The analysis of the bacterial diversity revealed a diverse infection status. Rickettsia was the most predominant endosymbiont while Cardinium was totally absent from all examined populations. Philaenus spumarius harbored Rickettsia, Arsenophonus, Hamiltonella and Wolbachia, N. campestris carried Rickettsia, Hamiltonella and Wolbachia while P. signatus was infected only by Rickettsia. The results of this study will provide an important knowledge resource for understanding the population dynamics of vectors of X. fastidiosa with a view to formulate effective management strategies towards the bacterium.}, } @article {pmid33572512, year = {2021}, author = {Hou, HX and Zhao, D and Xiao, JH and Huang, DW}, title = {Transcriptomic Analysis Reveals the Sexually Divergent Host-Wolbachia Interaction Patterns in a Fig Wasp.}, journal = {Microorganisms}, volume = {9}, number = {2}, pages = {}, pmid = {33572512}, issn = {2076-2607}, support = {96172158, 96173250, 91822294//the construction funds for the "Double First-Class" initiative for Nankai University/ ; 31830084, 31970440, 31672336//National Natural Science Foundation of China/ ; }, abstract = {Wolbachia are widely distributed in arthropods and nematodes, acquiring nutrients from the hosts, and inducing remarkable reproductive modulations on the hosts. To investigate the interaction of Wolbachia and insects, Wolbachia are often artificially eliminated from Wolbachia-infected hosts, which may produce negative effects of antibiotics. In the present study, based on the transcriptomic data of a fig wasp species Ceratosolen solmsi with two sibling lineages, one natively infected and the other noninfected with Wolbachia, we investigated the expression patterns of genes. The comparison results of differently expressed genes (DEGs) between Wolbachia infected and noninfected samples show that males have many more DEGs than females. The male unique upregulated genes are enriched in biological processes mainly related to biosynthesis, transport, positive regulation of I-kappaB kinase/NF-kappaB signaling, MAPK cascade, and pathogenesis; the male unique downregulated genes are enriched in biological processes mainly related to transport, oxidation-reduction, cellular responses to oxidative stress, lipid oxidation, cytoskeleton organization, actin filament-based process, and localization. In addition, for the Wolbachia's gene expression, the number of genes up-regulated in males is higher than that in females. The results revealed divergent patterns of the host-Wolbachia interactions between males and females in the fig wasp species.}, } @article {pmid33564174, year = {2021}, author = {Salje, J}, title = {Cells within cells: Rickettsiales and the obligate intracellular bacterial lifestyle.}, journal = {Nature reviews. Microbiology}, volume = {19}, number = {6}, pages = {375-390}, pmid = {33564174}, issn = {1740-1534}, support = {R21 AI144385/AI/NIAID NIH HHS/United States ; R56 AI148645/AI/NIAID NIH HHS/United States ; }, mesh = {Bacterial Physiological Phenomena ; Genome, Bacterial ; Humans ; Phylogeny ; Rickettsiales/classification/genetics/*physiology ; }, abstract = {The Rickettsiales are a group of obligate intracellular vector-borne Gram-negative bacteria that include many organisms of clinical and agricultural importance, including Anaplasma spp., Ehrlichia chaffeensis, Wolbachia, Rickettsia spp. and Orientia tsutsugamushi. This Review provides an overview of the current state of knowledge of the biology of these bacteria and their interactions with host cells, with a focus on pathogenic species or those that are otherwise important for human health. This includes a description of rickettsial genomics, bacterial cell biology, the intracellular lifestyles of Rickettsiales and the mechanisms by which they induce and evade the innate immune response.}, } @article {pmid33563832, year = {2021}, author = {Lindsey, ARI and Bhattacharya, T and Hardy, RW and Newton, ILG}, title = {Wolbachia and Virus Alter the Host Transcriptome at the Interface of Nucleotide Metabolism Pathways.}, journal = {mBio}, volume = {12}, number = {1}, pages = {}, pmid = {33563832}, issn = {2150-7511}, support = {P40 OD018537/OD/NIH HHS/United States ; R01 AI144430/AI/NIAID NIH HHS/United States ; R21 AI121849/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology/virology ; Female ; Gene Expression Profiling ; Host-Pathogen Interactions/genetics ; Male ; *Metabolic Networks and Pathways ; *Microbial Interactions ; Mosquito Vectors/microbiology/virology ; Nucleotides/genetics/*metabolism ; Symbiosis ; *Transcriptome ; Virus Diseases/virology ; Virus Replication ; Viruses/*pathogenicity ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is a maternally transmitted bacterium that manipulates arthropod and nematode biology in myriad ways. The Wolbachia strain colonizing Drosophila melanogaster creates sperm-egg incompatibilities and protects its host against RNA viruses, making it a promising tool for vector control. Despite successful trials using Wolbachia-transfected mosquitoes for dengue control, knowledge of how Wolbachia and viruses jointly affect insect biology remains limited. Using the Drosophila melanogaster model, transcriptomics and gene expression network analyses revealed pathways with altered expression and splicing due to Wolbachia colonization and virus infection. Included are metabolic pathways previously unknown to be important for Wolbachia-host interactions. Additionally, Wolbachia-colonized flies exhibit a dampened transcriptomic response to virus infection, consistent with early blocking of virus replication. Finally, using Drosophila genetics, we show that Wolbachia and expression of nucleotide metabolism genes have interactive effects on virus replication. Understanding the mechanisms of pathogen blocking will contribute to the effective development of Wolbachia-mediated vector control programs.IMPORTANCE Recently developed arbovirus control strategies leverage the symbiotic bacterium Wolbachia, which spreads in insect populations and blocks viruses from replicating. While this strategy has been successful, details of how this "pathogen blocking" works are limited. Here, we use a combination of virus infections, fly genetics, and transcriptomics to show that Wolbachia and virus interact at host nucleotide metabolism pathways.}, } @article {pmid33563818, year = {2021}, author = {Sicard, M and Namias, A and Perriat-Sanguinet, M and Carron, E and Unal, S and Altinli, M and Landmann, F and Weill, M}, title = {Cytoplasmic Incompatibility Variations in Relation with Wolbachia cid Genes Divergence in Culex pipiens.}, journal = {mBio}, volume = {12}, number = {1}, pages = {}, pmid = {33563818}, issn = {2150-7511}, mesh = {Animals ; Cell Line ; Centromere Protein A/*genetics ; Culex/*microbiology/physiology ; Cytoplasm/*physiology ; Cytosol/*microbiology ; Female ; Genetic Drift ; Heterocyclic Compounds, 2-Ring ; Host Specificity ; Male ; Phenotype ; Phylogeny ; Symbiosis ; Thiourea/analogs & derivatives ; Wolbachia/*genetics ; }, abstract = {In arthropods, Wolbachia endosymbionts induce conditional sterility, called cytoplasmic incompatibility (CI), resulting from embryonic lethality. CI penetrance (i.e., embryonic death rate) varies depending on host species and Wolbachia strains involved. All Culex pipiens mosquitoes are infected by the endosymbiotic alphaproteobacteria Wolbachia wPip. CI in Culex, characterized as a binary "compatible/incompatible" phenomenon, revealed an unparalleled diversity of patterns linked to the amplification-diversification of cidA and cidB genes. Here, we accurately studied CI penetrance variations in the light of cid genes divergence by generating a C. pipiens compatibility matrix between 11 lines hosting different phylogenetic wPip groups and exhibiting distinct cid gene repertoires. We showed, as expected, that crosses involving wPip from the same group were mostly compatible. In contrast, only 22% of the crosses involving different wPip groups were compatible, while 54% were fully incompatible. For the remaining 24% of the crosses, "intermediate" compatibilities were reported, and a cytological observation of the first zygotic division confirmed the occurrence of "canonical" CI phenotypes in a fraction of the eggs. Backcross experiments demonstrated that intermediate compatibilities were not linked to host genetic background but to the Wolbachia strains involved. This previously unstudied intermediate penetrance CI was more severe and frequent in crosses involving wPip-IV strains exhibiting cid variants markedly divergent from other wPip groups. Our data demonstrate that CI is not always a binary compatible/incompatible phenomenon in C. pipiens but that intermediate compatibilities putatively resulting from partial mismatch due to Cid proteins divergence exist in this species complex.IMPORTANCECulex pipiens mosquitoes are infected with wPip. These endosymbionts induce a conditional sterility called CI resulting from embryonic deaths, which constitutes a cornerstone for Wolbachia antivectorial methods. Recent studies revealed that (i) two genes, cidA and cidB, are central in Wolbachia-CI mechanisms, and (ii) compatibility versus incompatibility between mosquito lines depends on the wPip phylogenetic groups at play. Here, we studied CI variations in relation to wPip groups and cid genes divergence. We showed, as expected, that the crosses involving wPip from the same group were compatible. In contrast, 78% of the crosses involving different wPip groups were partially or fully incompatible. In such crosses, we reported defects during the first zygotic division, a hallmark of CI. We showed that CI was more severe and frequent in crosses involving wPip-IV strains exhibiting cid variants, which markedly diverge from those of other wPip groups.}, } @article {pmid33562305, year = {2021}, author = {Moo-Llanes, DA and López-Ordóñez, T and Torres-Monzón, JA and Mosso-González, C and Casas-Martínez, M and Samy, AM}, title = {Assessing the Potential Distributions of the Invasive Mosquito Vector Aedes albopictus and Its Natural Wolbachia Infections in México.}, journal = {Insects}, volume = {12}, number = {2}, pages = {}, pmid = {33562305}, issn = {2075-4450}, abstract = {The Asian tiger mosquito Aedes albopictus is currently the most invasive vector species, with a widespread global distribution. Aedes albopictus is the potential vector of diverse arboviruses, including Zika and dengue. This study updated the ecological niche model of Ae. albopictus and inferred the potential distribution of natural Wolbachia infections in Ae. albopictus in México. The ecological niche models were constructed based on diverse model settings to better estimate the potential distributions and uncertainty indices of both Ae. albopictus and its natural Wolbachia infections in México. The distribution of Ae. albopictus covered the states across Northern México, the Gulf of México, the Pacific Coast of México, Central México, and the southeast of México. The ecological niche model of the natural Wolbachia infections in Ae. albopictus populations anticipated the occurrence of natural Wolbachia infections in the southeast of México, the Chiapas border with Guatemala, and Veracruz. These results can be used to prioritize vector surveillance and control programs in México for strategic and future decision-making; however, it is still necessary to establish active surveillance programs to assess model predictions based on the independent sampling of Ae. albopictus from different invasion zones in México. Finally, vector surveillance should also screen the natural Wolbachia infections in Ae. albopictus to validate Wolbachia predictions across México, particularly in the southeast of México.}, } @article {pmid33555369, year = {2021}, author = {Dou, W and Miao, Y and Xiao, J and Huang, D}, title = {Association of Wolbachia with Gene Expression in Drosophila Testes.}, journal = {Microbial ecology}, volume = {82}, number = {3}, pages = {805-817}, pmid = {33555369}, issn = {1432-184X}, support = {31830084//National Natural Science Foundation of China/ ; 31970440//National Natural Science Foundation of China/ ; 31672336//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Drosophila ; Drosophila melanogaster/genetics ; Male ; Testis ; Transcriptome ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a genus of intracellular symbiotic bacteria that are widely distributed in arthropods and nematodes. These maternally inherited bacteria regulate host reproductive systems in various ways to facilitate their vertical transmission. Since the identification of Wolbachia in many insects, the relationship between Wolbachia and the host has attracted great interest. Numerous studies have indicated that Wolbachia modifies a variety of biological processes in the host. Previous studies in Drosophila melanogaster (D. melanogaster) have demonstrated that Wolbachia can affect spermatid differentiation, chromosome deposition, and sperm activity in the early stages of spermatogenesis, leading to sperm dysfunction. Here, we explored the putative effect of Wolbachia in sperm maturation using transcriptomic approaches to compare gene expression in Wolbachia-infected and Wolbachia-free D. melanogaster adult testes. Our findings show that Wolbachia affects many biological processes in D. melanogaster adult testes, and most of the differentially expressed genes involved in carbohydrate metabolism, lysosomal degradation, proteolysis, lipid metabolism, and immune response were upregulated in the presence of Wolbachia. In contrast, some genes that are putatively associated with cutin and wax biosynthesis and peroxisome pathways were downregulated. We did not find any differentially expressed genes that are predicted to be related to spermatogenesis in the datasets. This work provides additional information for understanding the Wolbachia-host intracellular relationships.}, } @article {pmid33555087, year = {2021}, author = {}, title = {Corrigendum: Wolbachia influence on the fitness of Anagyrus vladimiri (Hymenoptera: Encyrtidae), a bio-control agent of mealybugs.}, journal = {Pest management science}, volume = {}, number = {}, pages = {}, doi = {10.1002/ps.6316}, pmid = {33555087}, issn = {1526-4998}, } @article {pmid33542272, year = {2021}, author = {Sucháčková Bartoňová, A and Konvička, M and Marešová, J and Wiemers, M and Ignatev, N and Wahlberg, N and Schmitt, T and Faltýnek Fric, Z}, title = {Wolbachia affects mitochondrial population structure in two systems of closely related Palaearctic blue butterflies.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {3019}, pmid = {33542272}, issn = {2045-2322}, mesh = {Animals ; Butterflies/*genetics/microbiology/ultrastructure ; DNA, Mitochondrial/genetics ; Mitochondria/genetics/microbiology/*ultrastructure ; *Phylogeny ; Wolbachia/*genetics/pathogenicity ; }, abstract = {The bacterium Wolbachia infects many insect species and spreads by diverse vertical and horizontal means. As co-inherited organisms, these bacteria often cause problems in mitochondrial phylogeny inference. The phylogenetic relationships of many closely related Palaearctic blue butterflies (Lepidoptera: Lycaenidae: Polyommatinae) are ambiguous. We considered the patterns of Wolbachia infection and mitochondrial diversity in two systems: Aricia agestis/Aricia artaxerxes and the Pseudophilotes baton species complex. We sampled butterflies across their distribution ranges and sequenced one butterfly mitochondrial gene and two Wolbachia genes. Both butterfly systems had uninfected and infected populations, and harboured several Wolbachia strains. Wolbachia was highly prevalent in A. artaxerxes and the host's mitochondrial structure was shallow, in contrast to A. agestis. Similar bacterial alleles infected both Aricia species from nearby sites, pointing to a possible horizontal transfer. Mitochondrial history of the P. baton species complex mirrored its Wolbachia infection and not the taxonomical division. Pseudophilotes baton and P. vicrama formed a hybrid zone in Europe. Wolbachia could obscure mitochondrial history, but knowledge on the infection helps us to understand the observed patterns. Testing for Wolbachia should be routine in mitochondrial DNA studies.}, } @article {pmid33536644, year = {2021}, author = {Li, Y and Baton, LA and Zhang, D and Bouyer, J and Parker, AG and Hoffmann, AA and Ng, LC and Tan, CH and Xi, Z}, title = {Reply to: Issues with combining incompatible and sterile insect techniques.}, journal = {Nature}, volume = {590}, number = {7844}, pages = {E3-E5}, pmid = {33536644}, issn = {1476-4687}, mesh = {*Aedes ; Animals ; *Wolbachia ; }, } @article {pmid33536643, year = {2021}, author = {Moretti, R and Calvitti, M}, title = {Issues with combining incompatible and sterile insect techniques.}, journal = {Nature}, volume = {590}, number = {7844}, pages = {E1-E2}, pmid = {33536643}, issn = {1476-4687}, mesh = {*Aedes ; Animals ; *Wolbachia ; }, } @article {pmid33531619, year = {2021}, author = {Sandoval-Mojica, AF and Hunter, WB and Aishwarya, V and Bonilla, S and Pelz-Stelinski, KS}, title = {Antibacterial FANA oligonucleotides as a novel approach for managing the Huanglongbing pathosystem.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {2760}, pmid = {33531619}, issn = {2045-2322}, mesh = {Animals ; Arabinonucleotides/administration & dosage/genetics ; Cell Line ; Citrus/microbiology ; Drosophila ; Gene Expression Regulation, Bacterial/*drug effects ; Gene Silencing ; Hemiptera/drug effects/*microbiology ; Insect Vectors/drug effects/microbiology ; Oligonucleotides, Antisense/*administration & dosage/genetics ; Plant Diseases/microbiology/*prevention & control ; Rhizobiaceae/*drug effects/genetics/pathogenicity ; Symbiosis/drug effects/genetics ; }, abstract = {Candidatus Liberibacter asiaticus (CLas), a bacterium transmitted by the Asian citrus psyllid, Diaphorina citri, is the causal agent of citrus greening disease, or Huanglongbng (HLB). Currently, vector population suppression with insecticides and tree removal are the most effective strategies for managing the HLB pathosystem. In this study, we assessed the bactericidal capabilities of 2'-deoxy-2'-fluoro-D-arabinonucleic acid antisense oligonucleotides (FANA ASO) both in vitro and in vivo by (1) confirming their capacity to penetrate insect cells, (2) silencing bacterial essential genes, and (3) quantifying reductions in bacterial titer and D. citri transmission. We confirmed that FANA ASO are able to penetrate insect cells without the use of a delivery agent. Expression of an essential gene in the D. citri endosymbiont, Wolbachia (wDi), significantly decreased by 30% following incubation with a wDi-specific FANA ASO. Viability of isolated wDi cells also decreased in response to the FANA ASO treatment. Delivery of a CLas-specific FANA ASO to infected adult D. citri in feeding assays resulted in significant silencing of a CLas essential gene. CLas relative density and transmission were significantly lower among D. citri fed FANA ASO in diet compared to untreated insects. Root infusions of a CLas-specific FANA ASO into infected Citrus trees significantly reduced CLas titer during a 30-day trial. Our results suggest that FANA ASO targeting insect-transmitted plant bacteria or insect endosymbionts may be useful tool for integrated management of agricultural pathogens.}, } @article {pmid33507381, year = {2021}, author = {Fallon, AM}, title = {DNA recombination and repair in Wolbachia: RecA and related proteins.}, journal = {Molecular genetics and genomics : MGG}, volume = {296}, number = {2}, pages = {437-456}, pmid = {33507381}, issn = {1617-4623}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Proteins/genetics/metabolism ; *DNA Repair ; Host Specificity ; Multigene Family ; Nematoda/*microbiology ; Rec A Recombinases/*genetics/metabolism ; Recombination, Genetic ; Serine Endopeptidases/genetics ; Species Specificity ; Wolbachia/classification/*genetics/metabolism ; }, abstract = {Wolbachia is an obligate intracellular bacterium that has undergone extensive genomic streamlining in its arthropod and nematode hosts. Because the gene encoding the bacterial DNA recombination/repair protein RecA is not essential in Escherichia coli, abundant expression of this protein in a mosquito cell line persistently infected with Wolbachia strain wStri was unexpected. However, RecA's role in the lytic cycle of bacteriophage lambda provides an explanation for retention of recA in strains known to encode lambda-like WO prophages. To examine DNA recombination/repair capacities in Wolbachia, a systematic examination of RecA and related proteins in complete or nearly complete Wolbachia genomes from supergroups A, B, C, D, E, F, J and S was undertaken. Genes encoding proteins including RecA, RecF, RecO, RecR, RecG and Holliday junction resolvases RuvA, RuvB and RuvC are uniformly absent from Wolbachia in supergroup C and have reduced representation in supergroups D and J, suggesting that recombination and repair activities are compromised in nematode-associated Wolbachia, relative to strains that infect arthropods. An exception is filarial Wolbachia strain wMhie, assigned to supergroup F, which occurs in a nematode host from a poikilothermic lizard. Genes encoding LexA and error-prone polymerases are absent from all Wolbachia genomes, suggesting that the SOS functions induced by RecA-mediated activation of LexA do not occur, despite retention of genes encoding a few proteins that respond to LexA induction in E. coli. Three independent E. coli accessions converge on a single Wolbachia UvrD helicase, which interacts with mismatch repair proteins MutS and MutL, encoded in nearly all Wolbachia genomes. With the exception of MutL, which has been mapped to a eukaryotic association module in Phage WO, proteins involved in recombination/repair are uniformly represented by single protein annotations. Putative phage-encoded MutL proteins are restricted to Wolbachia supergroups A and B and show higher amino acid identity than chromosomally encoded MutL orthologs. This analysis underscores differences between nematode and arthropod-associated Wolbachia and describes aspects of DNA metabolism that potentially impact development of procedures for transformation and genetic manipulation of Wolbachia.}, } @article {pmid33500597, year = {2021}, author = {Zimmermann, BL and Cardoso, GM and Bouchon, D and Pezzi, PH and Palaoro, AV and Araujo, PB}, title = {Supergroup F Wolbachia in terrestrial isopods: Horizontal transmission from termites?.}, journal = {Evolutionary ecology}, volume = {35}, number = {2}, pages = {165-182}, pmid = {33500597}, issn = {0269-7653}, abstract = {UNLABELLED: Horizontal transmission between distantly related species has been used to explain how Wolbachia infect multiple species at astonishing rates despite the selection for resistance. Recently, a terrestrial isopod species was found to be infected by an unusual strain of supergroup F Wolbachia. However, only Wolbachia of supergroup B is typically found in isopods. One possibility is that these isopods acquired the infection because of their recurrent contact with termites-a group with strong evidence of infection by supergroup F Wolbachia. Thus, our goals were: (1) check if the infection was an isolated case in isopods, or if it revealed a broader pattern; (2) search for Wolbachia infection in the termites within Brazil; and (3) look for evidence consistent with horizontal transmission between isopods and termites. We collected Neotroponiscus terrestrial isopods and termites along the Brazilian coastal Atlantic forest. We sequenced and identified the Wolbachia strains found in these groups using coxA, dnaA, and fpbA genes. We constructed phylogenies for both bacteria and host taxa and tested for coevolution. We found the supergroup F Wolbachia in other species and populations of Neotroponiscus, and also in Nasutitermes and Procornitermes termites. The phylogenies showed that, despite the phylogenetic distance between isopods and termites, the Wolbachia strains clustered together. Furthermore, cophylogenetic analyses showed significant jumps of Wolbachia between terrestrial isopods and termites. Thus, our study suggests that the horizontal transmission of supergroup F Wolbachia between termites and terrestrial isopods is likely. Our study also helps understanding the success and worldwide distribution of this symbiont.

SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s10682-021-10101-4) contains supplementary material, which is available to authorized users.}, } @article {pmid33486127, year = {2021}, author = {Lozano-Sardaneta, YN and Valderrama, A and Sánchez-Montes, S and Grostieta, E and Colunga-Salas, P and Sánchez-Cordero, V and Becker, I}, title = {Rickettsial agents detected in the genus Psathyromyia (Diptera:Phlebotominae) from a Biosphere Reserve of Veracruz, Mexico.}, journal = {Parasitology international}, volume = {82}, number = {}, pages = {102286}, doi = {10.1016/j.parint.2021.102286}, pmid = {33486127}, issn = {1873-0329}, mesh = {Animals ; Female ; Mexico ; Psychodidae/*microbiology ; Rickettsia/*isolation & purification ; Wolbachia/*isolation & purification ; }, abstract = {Phlebotomine sand flies are considered the main vectors of Leishmania, the causal agents of leishmaniasis, which is a serious emerging public health problem worldwide. The use of biological control alternatives, like endosymbiotic bacteria (Wolbachia and Rickettsia), have been proposed to decrease sand fly populations and reduce Leishmania transmissions, yet only few records on the detection of Wolbachia or Rickettsia in sand flies are available worldwide. The aim of this study was to perform the molecular detection of Rickettsial agents associated with sand flies from the last patch of a rainforest in south-eastern Mexico, where a high prevalence of Leishmania infantum has been reported. Sampling effort of sand flies covered 300 trap-nights between 2011 and 2013, and a total of 925 specimens from twelve species were morphologically identified. Using PCR techniques, we identified a new lineage of the endosymbionts Rickettsia in Psathyromyia aclydifera (prevalence of 19.54%), and Wolbachia in Psathyromyia shannoni and Lutzomyia sp. (prevalence of 25%). The detected Wolbachia lineage was similar to the wWhi strain found in Pa. shannoni from Colombia and Nyssomyia whitmani from Brazil; whereas the identified Rickettsia represents a new lineage worldwide. This is the first record of Rickettsial agents associated to sand flies from this region, yet it remains for analysed if these bacteria possibly play a role as vector control agents, capable of reducing the sand fly populations in Mexico.}, } @article {pmid33484388, year = {2021}, author = {Yang, K and Chen, H and Bing, XL and Xia, X and Zhu, YX and Hong, XY}, title = {Wolbachia and Spiroplasma could influence bacterial communities of the spider mite Tetranychus truncatus.}, journal = {Experimental & applied acarology}, volume = {83}, number = {2}, pages = {197-210}, pmid = {33484388}, issn = {1572-9702}, support = {32020103011//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; 31901888//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Bacteria/genetics ; RNA, Ribosomal, 16S/genetics ; *Spiroplasma/genetics ; Symbiosis ; *Tetranychidae/genetics ; *Wolbachia/genetics ; }, abstract = {The structures of arthropod bacterial communities are complex. These microbiotas usually provide many beneficial services to their hosts, whereas occasionally they may be parasitical. To date, little is known about the bacterial communities of Tetranychus truncatus and the factors contributing to the structure of its bacterial communities are unexplored yet. Here, we used four symbiont-infected T. truncatus strains-including one Wolbachia and Spiroplasma co-infected strain, two symbiont singly-infected strains and one symbiont uninfected strain-to investigate the influence of endosymbionts on the structure of the host mites' microbiota. Based on 16S rRNA genes sequencing analysis, we found Wolbachia and Spiroplasma were the two most abundant bacteria in T. truncatus and the presence of both symbionts could not change the diversity of bacterial communities (based on alpha-diversity indexes such as ACE, Chao1, Shannon and Simpson diversity index). Symbiont infection did alter the abundance of many other bacterial genera, such as Megamonas and Bacteroides. The structures of bacterial communities differed significantly among symbiont-infected strains. These results suggested a prominent effect of Wolbachia and Spiroplasma on bacterial communities of the host T. truncatus. These findings advance our understanding of T. truncatus microbiota and will be helpful for further study on bacterial communities of spider mites.}, } @article {pmid33468006, year = {2021}, author = {Yoshida, K and Sanada-Morimura, S and Huang, SH and Tokuda, M}, title = {Silence of the killers: discovery of male-killing suppression in a rearing strain of the small brown planthopper, Laodelphax striatellus.}, journal = {Proceedings. Biological sciences}, volume = {288}, number = {1943}, pages = {20202125}, pmid = {33468006}, issn = {1471-2954}, mesh = {Animals ; Female ; *Hemiptera/genetics ; Male ; Sex Ratio ; *Spiroplasma ; *Wolbachia ; }, abstract = {According to evolutionary theory, sex ratio distortions caused by reproductive parasites such as Wolbachia and Spiroplasma are predicted to be rapidly normalized by the emergence of host nuclear suppressors. However, such processes in the evolutionary arms race are difficult to observe because sex ratio biases will be promptly hidden and become superficially unrecognizable. The evolution of genetic suppressors has been reported in just two insect species so far. In the small brown planthopper, Laodelphax striatellus, female-biases caused by Spiroplasma, which is a 'late' male-killer, have been found in some populations. During the continuous rearing of L. striatellus, we noted that a rearing strain had a 1 : 1 sex ratio even though it harboured Spiroplasma. Through introgression crossing experiments with a strain lacking suppressors, we revealed that the L. striatellus strain had the zygotic male-killing suppressor acting as a dominant trait. The male-killing phenotype was hidden by the suppressor even though Spiroplasma retained its male-killing ability. This is the first study to demonstrate the existence of a late male-killing suppressor and its mode of inheritance. Our results, together with those of previous studies, suggest that the inheritance modes of male-killing suppressors are similar regardless of insect order or early or late male killing.}, } @article {pmid33466915, year = {2021}, author = {Dong, S and Dimopoulos, G}, title = {Antiviral Compounds for Blocking Arboviral Transmission in Mosquitoes.}, journal = {Viruses}, volume = {13}, number = {1}, pages = {}, pmid = {33466915}, issn = {1999-4915}, support = {R01 AI141532/AI/NIAID NIH HHS/United States ; R01AI141532/NH/NIH HHS/United States ; }, mesh = {Aedes/drug effects/*virology ; Animals ; Antiviral Agents/chemistry/*pharmacology/therapeutic use ; Arbovirus Infections/drug therapy/*transmission/*virology ; Arboviruses/classification/*drug effects ; Cells, Cultured ; Drug Discovery/methods ; Drug Evaluation, Preclinical ; Humans ; Mosquito Control/methods ; Mosquito Vectors/*virology ; Vector Borne Diseases/drug therapy/transmission/virology ; Virus Replication/drug effects ; }, abstract = {Mosquito-borne arthropod-borne viruses (arboviruses) such as the dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) are important human pathogens that are responsible for significant global morbidity and mortality. The recent emergence and re-emergence of mosquito-borne viral diseases (MBVDs) highlight the urgent need for safe and effective vaccines, therapeutics, and vector-control approaches to prevent MBVD outbreaks. In nature, arboviruses circulate between vertebrate hosts and arthropod vectors; therefore, disrupting the virus lifecycle in mosquitoes is a major approach for combating MBVDs. Several strategies were proposed to render mosquitoes that are refractory to arboviral infection, for example, those involving the generation of genetically modified mosquitoes or infection with the symbiotic bacterium Wolbachia. Due to the recent development of high-throughput screening methods, an increasing number of drugs with inhibitory effects on mosquito-borne arboviruses in mammalian cells were identified. These antivirals are useful resources that can impede the circulation of arboviruses between arthropods and humans by either rendering viruses more vulnerable in humans or suppressing viral infection by reducing the expression of host factors in mosquitoes. In this review, we summarize recent advances in small-molecule antiarboviral drugs in mammalian and mosquito cells, and discuss how to use these antivirals to block the transmission of MBVDs.}, } @article {pmid33454808, year = {2021}, author = {Takano, SI and Gotoh, Y and Hayashi, T}, title = {"Candidatus Mesenet longicola": Novel Endosymbionts of Brontispa longissima that Induce Cytoplasmic Incompatibility.}, journal = {Microbial ecology}, volume = {82}, number = {2}, pages = {512-522}, pmid = {33454808}, issn = {1432-184X}, support = {18H02207//Japan Society for the Promotion of Science/ ; 16H06279//Japan Society for the Promotion of Science/ ; }, mesh = {Animals ; *Coleoptera ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Intracellular bacteria that are mainly transmitted maternally affect their arthropod hosts' biology in various ways. One such effect is known as cytoplasmic incompatibility (CI), and three bacterial species are known to induce CI: Wolbachia, Cardinium hertigii, and a recently found alphaproteobacterial symbiont. To clarify the taxonomic status and provide the foundation for future studies to reveal CI mechanisms and other phenotypes, we investigated genetic and morphological properties of the third CI inducer that we have previously reported inducing CI in the coconut beetle Brontispa longissima. The draft genome of the bacteria was obtained from the oocytes of two isofemale lines of B. longissima infected with the bacteria: one from Japan (GL2) and the other from Vietnam (L5). Genome features of the symbionts (sGL2 and sL5) were highly similar, showing 1.3 Mb in size, 32.1% GC content, and 99.83% average nucleotide sequence. A phylogenetic study based on 43 universal and single-copy phylogenetic marker genes indicates that they formed a distinct clade in the family Anaplasmataceae. 16S rRNA gene sequences indicate that they are different from the closest known relatives, at least at the genus level. Therefore, we propose a new genus and species, "Candidatus Mesenet longicola", for the symbionts of B. longissima. Morphological analyses showed that Ca. M. longicola is an intracellular bacterium that is ellipsoidal to rod-shaped and 0.94 ± 0.26 μm (mean ± SD) in length, and accumulated in the anterior part of the oocyte. Candidates for the Ca. M. longicola genes responsible for CI induction are also described.}, } @article {pmid33452445, year = {2021}, author = {Santos, LMB and Mutsaers, M and Garcia, GA and David, MR and Pavan, MG and Petersen, MT and Corrêa-Antônio, J and Couto-Lima, D and Maes, L and Dowell, F and Lord, A and Sikulu-Lord, M and Maciel-de-Freitas, R}, title = {High throughput estimates of Wolbachia, Zika and chikungunya infection in Aedes aegypti by near-infrared spectroscopy to improve arbovirus surveillance.}, journal = {Communications biology}, volume = {4}, number = {1}, pages = {67}, pmid = {33452445}, issn = {2399-3642}, mesh = {Aedes/*microbiology/*virology ; Animals ; Bacterial Infections/diagnosis/veterinary ; Chikungunya Fever/diagnosis/veterinary ; Female ; High-Throughput Screening Assays/methods ; Spectroscopy, Near-Infrared/*methods ; Time Factors ; Wolbachia ; Zika Virus Infection/diagnosis/veterinary ; }, abstract = {Deployment of Wolbachia to mitigate dengue (DENV), Zika (ZIKV) and chikungunya (CHIKV) transmission is ongoing in 12 countries. One way to assess the efficacy of Wolbachia releases is to determine invasion rates within the wild population of Aedes aegypti following their release. Herein we evaluated the accuracy, sensitivity and specificity of the Near Infrared Spectroscopy (NIRS) in estimating the time post death, ZIKV-, CHIKV-, and Wolbachia-infection in trapped dead female Ae. aegypti mosquitoes over a period of 7 days. Regardless of the infection type, time post-death of mosquitoes was accurately predicted into four categories (fresh, 1 day old, 2-4 days old and 5-7 days old). Overall accuracies of 93.2, 97 and 90.3% were observed when NIRS was used to detect ZIKV, CHIKV and Wolbachia in dead Ae. aegypti female mosquitoes indicating NIRS could be potentially applied as a rapid and cost-effective arbovirus surveillance tool. However, field data is required to demonstrate the full capacity of NIRS for detecting these infections under field conditions.}, } @article {pmid33441735, year = {2021}, author = {Díaz-Nieto, LM and Gil, MF and Lazarte, JN and Perotti, MA and Berón, CM}, title = {Culex quinquefasciatus carrying Wolbachia is less susceptible to entomopathogenic bacteria.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {1094}, pmid = {33441735}, issn = {2045-2322}, mesh = {Animals ; Culex/*microbiology/physiology ; Female ; Host-Pathogen Interactions ; Male ; *Pest Control, Biological ; Wolbachia/*physiology ; }, abstract = {In an attempt to evaluate the susceptibility of the mosquito Culex quinquefasciatus to bacterial agents, a population naturally infected with a Wolbachia pipientis wPipSJ native strain was tested against the action of three bacterial mosquitocides, Bacillus thuringiensis subsp. israelensis, Bacillus wiedmannii biovar thuringiensis and Lysinibacillus sphaericus. Tests were carried out on mosquito larvae with and without Wolbachia (controls). Cx. quinquefasciatus naturally infected with the native wPipSJ strain proved to be more resistant to the pathogenic action of the three mosquitocidal bacterial strains. Additionally, wPipSJ was fully characterised using metagenome-assembled genomics, PCR-RFLP (PCR-Restriction Fragment Length Polymorphism) and MLST (MultiLocus Sequence Typing) analyses. This Wolbachia strain wPipSJ belongs to haplotype I, group wPip-III and supergroup B, clustering with other mosquito wPip strains, such as wPip PEL, wPip JHB, wPip Mol, and wAlbB; showing the southernmost distribution in America. The cytoplasmic incompatibility phenotype of this strain was revealed via crosses between wildtype (Wolbachia[+]) and antibiotic treated mosquito populations. The results of the tests with the bacterial agents suggest that Cx. quinquefasciatus naturally infected with wPipSJ is less susceptible to the pathogenic action of mosquitocidal bacterial strains when compared with the antibiotic-treated mosquito isoline, and is more susceptible to B. thuringiensis subsp. israelensis than to the other two mosquitocidal agents.}, } @article {pmid33436511, year = {2021}, author = {Chung, M and Adkins, RS and Mattick, JSA and Bradwell, KR and Shetty, AC and Sadzewicz, L and Tallon, LJ and Fraser, CM and Rasko, DA and Mahurkar, A and Dunning Hotopp, JC}, title = {FADU: a Quantification Tool for Prokaryotic Transcriptomic Analyses.}, journal = {mSystems}, volume = {6}, number = {1}, pages = {}, pmid = {33436511}, issn = {2379-5077}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {Quantification tools for RNA sequencing (RNA-Seq) analyses are often designed and tested using human transcriptomics data sets, in which full-length transcript sequences are well annotated. For prokaryotic transcriptomics experiments, full-length transcript sequences are seldom known, and coding sequences must instead be used for quantification steps in RNA-Seq analyses. However, operons confound accurate quantification of coding sequences since a single transcript does not necessarily equate to a single gene. Here, we introduce FADU (Feature Aggregate Depth Utility), a quantification tool designed specifically for prokaryotic RNA-Seq analyses. FADU assigns partial count values proportional to the length of the fragment overlapping the target feature. To assess the ability of FADU to quantify genes in prokaryotic transcriptomics analyses, we compared its performance to those of eXpress, featureCounts, HTSeq, kallisto, and Salmon across three paired-end read data sets of (i) Ehrlichia chaffeensis, (ii) Escherichia coli, and (iii) the Wolbachia endosymbiont wBm. Across each of the three data sets, we find that FADU can more accurately quantify operonic genes by deriving proportional counts for multigene fragments within operons. FADU is available at https://github.com/IGS/FADUIMPORTANCE Most currently available quantification tools for transcriptomics analyses have been designed for human data sets, in which full-length transcript sequences, including the untranslated regions, are well annotated. In most prokaryotic systems, full-length transcript sequences have yet to be characterized, leading to prokaryotic transcriptomics analyses being performed based on only the coding sequences. In contrast to eukaryotes, prokaryotes contain polycistronic transcripts, and when genes are quantified based on coding sequences instead of transcript sequences, this leads to an increased abundance of improperly assigned ambiguous multigene fragments, specifically those mapping to multiple genes in operons. Here, we describe FADU, a quantification tool for prokaryotic RNA-Seq analyses designed to assign proportional counts with the purpose of better quantifying operonic genes while minimizing the pitfalls associated with improperly assigning fragment counts from ambiguous transcripts.}, } @article {pmid33436431, year = {2021}, author = {Deehan, M and Lin, W and Blum, B and Emili, A and Frydman, H}, title = {Intracellular Density of Wolbachia Is Mediated by Host Autophagy and the Bacterial Cytoplasmic Incompatibility Gene cifB in a Cell Type-Dependent Manner in Drosophila melanogaster.}, journal = {mBio}, volume = {12}, number = {1}, pages = {}, pmid = {33436431}, issn = {2150-7511}, mesh = {Animals ; Autophagy/*genetics/physiology ; Autophagy-Related Protein-1 Homolog ; Bacterial Proteins/metabolism ; Cytoplasm/*microbiology ; Cytosol ; Drosophila Proteins ; Drosophila melanogaster/genetics/immunology/*microbiology ; Female ; Gene Knockdown Techniques ; Genes, Bacterial/*genetics ; Host-Pathogen Interactions/*genetics/immunology ; Immunity, Innate ; Male ; Reproduction ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Autophagy is an intracellular degradation pathway involved in innate immunity. Pathogenic bacteria have evolved several mechanisms to escape degradation or exploit autophagy to acquire host nutrients. In the case of endosymbionts, which often have commensal or mutualistic interactions with the host, autophagy is not well characterized. We utilized tissue-specific autophagy mutants to determine if Wolbachia, a vertically transmitted obligate endosymbiont of Drosophila melanogaster, is regulated by autophagy in somatic and germ line cell types. Our analysis revealed core autophagy proteins Atg1 and Atg8 and a selective autophagy-specific protein Ref(2)p negatively regulate Wolbachia in the hub, a male gonad somatic cell type. Furthermore, we determined that the Wolbachia effector protein, CifB, modulates autophagy-Wolbachia interactions, identifying a new host-related pathway which these bacterial proteins interact with. In the female germ line, the cell type necessary for inheritance of Wolbachia through vertical transmission, we discovered that bulk autophagy mediated by Atg1 and Atg8 positively regulates Wolbachia density, whereas Ref(2)p had no effect. Global metabolomics of fly ovaries deficient in germ line autophagy revealed reduced lipid and carbon metabolism, implicating metabolites from these pathways as positive regulators of Wolbachia Our work provides further understanding of how autophagy affects bacteria in a cell type-dependent manner.IMPORTANCE Autophagy is a eukaryotic intracellular degradation pathway which can act as an innate immune response to eliminate pathogens. Conversely, pathogens can evolve proteins which modulate the autophagy pathway to subvert degradation and establish an infection. Wolbachia, a vertically transmitted obligate endosymbiont which infects up to 40% of insect species, is negatively regulated by autophagy in whole animals, but the specific molecular mechanism and tissue which govern this interaction remain unknown. Our studies use cell type-specific autophagy mutants to reveal that Wolbachia is negatively regulated by selective autophagy in the soma, while nonselective autophagy positively regulates Wolbachia in the female germ line. These data provide evidence that cell type can drive different basal autophagy programs which modulate intracellular microbes differently. Additionally, we identified that the Wolbachia effector CifB acts in the selective autophagy pathway to aid in intracellular bacterial survival, providing a new function for CifB beyond its previously identified role in reproductive manipulation.}, } @article {pmid33435566, year = {2021}, author = {Ogunlade, ST and Meehan, MT and Adekunle, AI and Rojas, DP and Adegboye, OA and McBryde, ES}, title = {A Review: Aedes-Borne Arboviral Infections, Controls and Wolbachia-Based Strategies.}, journal = {Vaccines}, volume = {9}, number = {1}, pages = {}, pmid = {33435566}, issn = {2076-393X}, abstract = {Arthropod-borne viruses (Arboviruses) continue to generate significant health and economic burdens for people living in endemic regions. Of these viruses, some of the most important (e.g., dengue, Zika, chikungunya, and yellow fever virus), are transmitted mainly by Aedes mosquitoes. Over the years, viral infection control has targeted vector population reduction and inhibition of arboviral replication and transmission. This control includes the vector control methods which are classified into chemical, environmental, and biological methods. Some of these control methods may be largely experimental (both field and laboratory investigations) or widely practised. Perceptively, one of the biological methods of vector control, in particular, Wolbachia-based control, shows a promising control strategy for eradicating Aedes-borne arboviruses. This can either be through the artificial introduction of Wolbachia, a naturally present bacterium that impedes viral growth in mosquitoes into heterologous Aedes aegypti mosquito vectors (vectors that are not natural hosts of Wolbachia) thereby limiting arboviral transmission or via Aedes albopictus mosquitoes, which naturally harbour Wolbachia infection. These strategies are potentially undermined by the tendency of mosquitoes to lose Wolbachia infection in unfavourable weather conditions (e.g., high temperature) and the inhibitory competitive dynamics among co-circulating Wolbachia strains. The main objective of this review was to critically appraise published articles on vector control strategies and specifically highlight the use of Wolbachia-based control to suppress vector population growth or disrupt viral transmission. We retrieved studies on the control strategies for arboviral transmissions via arthropod vectors and discussed the use of Wolbachia control strategies for eradicating arboviral diseases to identify literature gaps that will be instrumental in developing models to estimate the impact of these control strategies and, in essence, the use of different Wolbachia strains and features.}, } @article {pmid33424811, year = {2020}, author = {Thongprem, P and Evison, SEF and Hurst, GDD and Otti, O}, title = {Transmission, Tropism, and Biological Impacts of Torix Rickettsia in the Common Bed Bug Cimex lectularius (Hemiptera: Cimicidae).}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {608763}, pmid = {33424811}, issn = {1664-302X}, abstract = {The torix group of Rickettsia have been recorded from a wide assemblage of invertebrates, but details of transmission and biological impacts on the host have rarely been established. The common bed bug (Cimex lectularius) is a hemipteran insect which lives as an obligatory hematophagous pest of humans and is host to a primary Wolbachia symbiont and two facultative symbionts, a BEV-like symbiont, and a torix group Rickettsia. In this study, we first note the presence of a single Rickettsia strain in multiple laboratory bed bug isolates derived from Europe and Africa. Importantly, we discovered that the Rickettsia has segregated in two laboratory strains, providing infected and uninfected isogenic lines for study. Crosses with these lines established transmission was purely maternal. Fluorescence in-situ hybridization analysis indicates Rickettsia infection in oocytes, bacteriomes, and other somatic tissues. We found no evidence that Rickettsia infection was associated with sex ratio distortion activity, but Rickettsia infected individuals developed from first instar to adult more slowly. The impact of Rickettsia on fecundity and fertility resulted in infected females producing fewer fertile eggs. However, we could not find any evidence for cytoplasmic incompatibility associated with Rickettsia presence. These data imply the existence of an unknown benefit to C. lectularius carrying Rickettsia that awaits further research.}, } @article {pmid33424808, year = {2020}, author = {Doremus, MR and Stouthamer, CM and Kelly, SE and Schmitz-Esser, S and Hunter, MS}, title = {Cardinium Localization During Its Parasitoid Wasp Host's Development Provides Insights Into Cytoplasmic Incompatibility.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {606399}, pmid = {33424808}, issn = {1664-302X}, abstract = {Arthropods harbor heritable intracellular symbionts that may manipulate host reproduction to favor symbiont transmission. In cytoplasmic incompatibility (CI), the symbiont sabotages the reproduction of infected males such that high levels of offspring mortality result when they mate with uninfected females. In crosses with infected males and infected females, however (the "rescue" cross), normal numbers of offspring are produced. A common CI-inducing symbiont, Cardinium hertigii, causes variable levels of CI mortality in the parasitoid wasp, Encarsia suzannae. Previous work correlated CI-induced mortality with male development time in this system, although the timing of Cardinium CI-induction and the relationship between development time and CI mortality was not well understood. Here, using a combination of crosses, manipulation of development time, and fluorescence microscopy, we identify the localization and the timing of the CI-induction step in the Cardinium-E. suzannae system. Antibiotic treatment of adult Cardinium-infected males did not reduce the mortality associated with the CI phenotype, suggesting that CI-alteration occurs prior to adulthood. Our results suggest that the alteration step occurs during the pupal period, and is limited by the duration of pupal development: 1) Encarsia produces most sperm prior to adulthood, 2) FISH localization of Cardinium in testes showed an association with sperm nuclei throughout spermatogenesis but not with mature sperm, and 3) two methods of prolonging the pupal period (cool temperatures and the juvenile hormone analog methoprene) both caused greater CI mortality, suggesting the degree of alteration is limited by the duration of the pupal stage. Based on these results, we compare two models for potential mechanisms of Cardinium sperm modification in the context of what is known about analogous mechanisms of Wolbachia, a more extensively studied CI-inducing symbiont.}, } @article {pmid33419044, year = {2021}, author = {Inácio da Silva, LM and Dezordi, FZ and Paiva, MHS and Wallau, GL}, title = {Systematic Review of Wolbachia Symbiont Detection in Mosquitoes: An Entangled Topic about Methodological Power and True Symbiosis.}, journal = {Pathogens (Basel, Switzerland)}, volume = {10}, number = {1}, pages = {}, pmid = {33419044}, issn = {2076-0817}, abstract = {Wolbachia is an endosymbiotic bacterium that naturally infects several arthropods and nematode species. Wolbachia gained particular attention due to its impact on their host fitness and the capacity of specific Wolbachia strains in reducing pathogen vector and agricultural pest populations and pathogens transmission. Despite the success of mosquito/pathogen control programs using Wolbachia-infected mosquito release, little is known about the abundance and distribution of Wolbachia in most mosquito species, a crucial knowledge for planning and deployment of mosquito control programs and that can further improve our basic biology understanding of Wolbachia and host relationships. In this systematic review, Wolbachia was detected in only 30% of the mosquito species investigated. Fourteen percent of the species were considered positive by some studies and negative by others in different geographical regions, suggesting a variable infection rate and/or limitations of the Wolbachia detection methods employed. Eighty-three percent of the studies screened Wolbachia with only one technique. Our findings highlight that the assessment of Wolbachia using a single approach limited the inference of true Wolbachia infection in most of the studied species and that researchers should carefully choose complementary methodologies and consider different Wolbachia-mosquito population dynamics that may be a source of bias to ascertain the correct infectious status of the host species.}, } @article {pmid33413518, year = {2021}, author = {Shults, P and Cohnstaedt, LW and Adelman, ZN and Brelsfoard, C}, title = {Next-generation tools to control biting midge populations and reduce pathogen transmission.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {31}, pmid = {33413518}, issn = {1756-3305}, mesh = {Animals ; Ceratopogonidae/genetics/microbiology/*parasitology ; Ecosystem ; Insect Control/*methods ; Insect Vectors/genetics/microbiology/*parasitology ; Larva/genetics/microbiology/parasitology ; Livestock/parasitology ; Wolbachia/pathogenicity ; }, abstract = {Biting midges of the genus Culicoides transmit disease-causing agents resulting in a significant economic impact on livestock industries in many parts of the world. Localized control efforts, such as removal of larval habitat or pesticide application, can be logistically difficult, expensive and ineffective if not instituted and maintained properly. With these limitations, a population-level approach to the management of Culicoides midges should be investigated as a means to replace or supplement existing control strategies. Next-generation control methods such as Wolbachia- and genetic-based population suppression and replacement are being investigated in several vector species. Here we assess the feasibility and applicability of these approaches for use against biting midges. We also discuss the technical and logistical hurdles needing to be addressed for each method to be successful, as well as emphasize the importance of addressing community engagement and involving stakeholders in the investigation and development of these approaches.}, } @article {pmid33407798, year = {2021}, author = {Farnesi, LC and Carvalho, FD and Lacerda, APC and Moreira, LA and Bruno, RV}, title = {The influence of different sources of blood meals on the physiology of Aedes aegypti harboring Wolbachia wMel: mouse blood as an alternative for mosquito rearing.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {21}, pmid = {33407798}, issn = {1756-3305}, mesh = {*Aedes/growth & development/microbiology/physiology ; Animal Feed ; Animals ; Arbovirus Infections/transmission ; *Blood ; Disease Vectors ; Fertility ; Guinea Pigs ; Humans ; Insect Control ; Longevity ; Mice ; Mosquito Vectors/microbiology/physiology ; Pest Control, Biological/methods ; Reproduction ; *Wolbachia ; }, abstract = {BACKGROUND: Aedes aegypti control programs have failed to restrain mosquito population expansion and, consequently, the spread of diseases such as dengue, Zika, and Chikungunya. Wolbachia infection of mosquitoes is a new and promising complementary tool for the control of arbovirus transmission. The use of Wolbachia-infected mosquitoes, mass reared using human blood, is currently being tested in several countries. However, the use of human blood for mass rearing mosquitoes, and thus expansion of this strategy, is problematic. With the aim of overcoming this problem, we tested the effect of different types of blood source on the fitness parameters of female Ae. aegypti and the Wolbachia titer over generations to be able to guarantee the suitability of an alternative source to human blood for mass rearing Wolbachia-infected mosquitoes.

METHODS: We investigated and compared essential parameters of the vector capacity of laboratory strains of Ae. aegypti with and without Wolbachia that fed on blood of different types of host (human, guinea pig, and mouse). The parameters analyzed were fecundity, fertility, pupation dynamics, and adult survival. Also, we tested whether it is possible to maintain mosquitoes with Wolbachia on mouse blood over generations without losing the bacterium titer.

RESULTS: The average number of eggs per female, egg viability and pupation dynamics in the Wolbachia-infected mosquito (wMelBr) strain were similar, regardless of the blood source. The F1 progenies of females that fed on mouse blood or human blood were analyzed. The longevity of males was lower than that of females. F1 female survival differed depending on the presence of Wolbachia in the mother. In subsequent generations analyzed up until F35, the relative Wolbachia density was even higher when mosquitoes fed on mouse blood in comparison to human blood.

CONCLUSIONS: Taken together, our results provide no evidence that the different types of blood influenced the fitness of the Wolbachia-infected mosquitoes. The presence of the bacterium in the colonies of Wolbachia-infected Ae. aegypti after 35 generations under the conditions evaluated indicates that they can be maintained on mouse blood. Based on these results, we show that it is possible to use mouse blood to feed female mosquitoes when using human blood for this purpose is problematic.}, } @article {pmid33406151, year = {2021}, author = {Chevignon, G and Foray, V and Pérez-Jiménez, MM and Libro, S and Chung, M and Foster, JM and Landmann, F}, title = {Dual RNAseq analyses at soma and germline levels reveal evolutionary innovations in the elephantiasis-agent Brugia malayi, and adaptation of its Wolbachia endosymbionts.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {1}, pages = {e0008935}, pmid = {33406151}, issn = {1935-2735}, mesh = {Animals ; *Biological Evolution ; Brugia malayi/*genetics ; Caenorhabditis elegans ; *Carisoprodol ; Elephantiasis/*genetics ; Elephantiasis, Filarial/genetics ; Female ; Gene Expression ; Genome ; *Germ Cells ; Humans ; Oogenesis ; Sequence Analysis, RNA ; Symbiosis ; Wolbachia/physiology ; }, abstract = {Brugia malayi is a human filarial nematode responsible for elephantiasis, a debilitating condition that is part of a broader spectrum of diseases called filariasis, including lymphatic filariasis and river blindness. Almost all filarial nematode species infecting humans live in mutualism with Wolbachia endosymbionts, present in somatic hypodermal tissues but also in the female germline which ensures their vertical transmission to the nematode progeny. These α-proteobacteria potentially provision their host with essential metabolites and protect the parasite against the vertebrate immune response. In the absence of Wolbachia wBm, B. malayi females become sterile, and the filarial nematode lifespan is greatly reduced. In order to better comprehend this symbiosis, we investigated the adaptation of wBm to the host nematode soma and germline, and we characterized these cellular environments to highlight their specificities. Dual RNAseq experiments were performed at the tissue-specific and ovarian developmental stage levels, reaching the resolution of the germline mitotic proliferation and meiotic differentiation stages. We found that most wBm genes, including putative effectors, are not differentially regulated between infected tissues. However, two wBm genes involved in stress responses are upregulated in the hypodermal chords compared to the germline, indicating that this somatic tissue represents a harsh environment to which wBm have adapted. A comparison of the B. malayi and C. elegans germline transcriptomes reveals a poor conservation of genes involved in the production of oocytes, with the filarial germline proliferative zone relying on a majority of genes absent from C. elegans. The first orthology map of the B. malayi genome presented here, together with tissue-specific expression enrichment analyses, indicate that the early steps of oogenesis are a developmental process involving genes specific to filarial nematodes, that likely result from evolutionary innovations supporting the filarial parasitic lifestyle.}, } @article {pmid33399931, year = {2021}, author = {Shih, CM and Ophine, L and Chao, LL}, title = {Molecular Detection and Genetic Identification of Wolbachia Endosymbiont in Wild-Caught Culex quinquefasciatus (Diptera: Culicidae) Mosquitoes from Sumatera Utara, Indonesia.}, journal = {Microbial ecology}, volume = {81}, number = {4}, pages = {1064-1074}, pmid = {33399931}, issn = {1432-184X}, support = {MOST 109-2314-B-037-077//Ministry of Science and Technology, Taiwan/ ; }, mesh = {Animals ; *Culex ; *Culicidae ; Female ; Indonesia ; Male ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {The genetic identity of Wolbachia endosymbiont in wild-caught Culex quinquefasciatus was determined for the first time in Indonesia. A total of 314 Cx. quinquefasciatus were examined for Wolbachia by PCR assay targeting the Wolbachia surface protein (wsp) gene. The prevalence of Wolbachia infection was detected in 29.94% of Cx. specimens (45.86% female and 8.27% male). The group-specific infection was detected with an infection rate of 0.32%, 28.98%, and 0.64% in groups A, B, and A&B, respectively. Phylogenetic analysis revealed all Wolbachia strains from Indonesia were genetically affiliated to the supergroup A and B with the high sequence similarity of 97.9-100% and 99.7-100%, respectively. Phylogenetic relationships can be easily distinguished by neighbor-joining analysis and were congruent by maximum likelihood method. The genetic distance (GD) values of intra- and inter-group analysis indicated a lower level (GD < 0.007 for group A and GD < 0.003 for group B) within the Indonesia strains and a higher level (GD > 1.125 for group A and GD > 1.129 for group B) as compared with other Wolbachia strains. Our results provide the first genetic identification of Wolbachia endosymbiont in Cx. quinquefasciatus collected from Indonesia, and the phylogenetic analysis revealed a new discovery of group A Wolbachia in wild-caught Cx. quinquefasciatus mosquitoes.}, } @article {pmid35526996, year = {2021}, author = {Fujita, R}, title = {[Osugoroshi virus, a male-killer virus].}, journal = {Uirusu}, volume = {71}, number = {1}, pages = {63-70}, doi = {10.2222/jsv.71.63}, pmid = {35526996}, issn = {0042-6857}, mesh = {Animals ; DNA Viruses ; Male ; *Moths/microbiology ; *RNA Viruses ; Sex Ratio ; *Spiroplasma ; Symbiosis ; *Viruses, Unclassified ; *Wolbachia ; }, abstract = {In insects, sex ratio bias is sometimes introduced by feminization, parthenogenesis, cytoplasmic incompatibility, or male-killing. Some intracellular bacteria such as Wolbachia or Spiroplasma has been known as male-killing agents. Here I introduce an example of non-bacterial male-killing agent, Osugoroshi virus found in oriental tea tortrix.}, } @article {pmid33362982, year = {2020}, author = {Driscoll, TP and Verhoeve, VI and Brockway, C and Shrewsberry, DL and Plumer, M and Sevdalis, SE and Beckmann, JF and Krueger, LM and Macaluso, KR and Azad, AF and Gillespie, JJ}, title = {Evolution of Wolbachia mutualism and reproductive parasitism: insight from two novel strains that co-infect cat fleas.}, journal = {PeerJ}, volume = {8}, number = {}, pages = {e10646}, pmid = {33362982}, issn = {2167-8359}, support = {R01 AI017828/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachiae are obligate intracellular bacteria that infect arthropods and certain nematodes. Usually maternally inherited, they may provision nutrients to (mutualism) or alter sexual biology of (reproductive parasitism) their invertebrate hosts. We report the assembly of closed genomes for two novel wolbachiae, wCfeT and wCfeJ, found co-infecting cat fleas (Ctenocephalides felis) of the Elward Laboratory colony (Soquel, CA, USA). wCfeT is basal to nearly all described Wolbachia supergroups, while wCfeJ is related to supergroups C, D and F. Both genomes contain laterally transferred genes that inform on the evolution of Wolbachia host associations. wCfeT carries the Biotin synthesis Operon of Obligate intracellular Microbes (BOOM); our analyses reveal five independent acquisitions of BOOM across the Wolbachia tree, indicating parallel evolution towards mutualism. Alternately, wCfeJ harbors a toxin-antidote operon analogous to the wPip cinAB operon recently characterized as an inducer of cytoplasmic incompatibility (CI) in flies. wCfeJ cinB and three adjacent genes are collectively similar to large modular toxins encoded in CI-like operons of certain Wolbachia strains and Rickettsia species, signifying that CI toxins streamline by fission of large modular toxins. Remarkably, the C. felis genome itself contains two CI-like antidote genes, divergent from wCfeJ cinA, revealing episodic reproductive parasitism in cat fleas and evidencing mobility of CI loci independent of WO-phage. Additional screening revealed predominant co-infection (wCfeT/wCfeJ) amongst C. felis colonies, though fleas in wild populations mostly harbor wCfeT alone. Collectively, genomes of wCfeT, wCfeJ, and their cat flea host supply instances of lateral gene transfers that could drive transitions between parasitism and mutualism.}, } @article {pmid33357053, year = {2021}, author = {Leftwich, PT and Spurgin, LG and Harvey-Samuel, T and Thomas, CJE and Paladino, LC and Edgington, MP and Alphey, L}, title = {Genetic pest management and the background genetics of release strains.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {376}, number = {1818}, pages = {20190805}, pmid = {33357053}, issn = {1471-2970}, support = {/WT_/Wellcome Trust/United Kingdom ; BBS/E/I/00007033/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 110117/Z/15/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Animals, Genetically Modified/genetics ; *Genetic Introgression ; *Insecta/genetics ; Pest Control, Biological/*instrumentation ; Reproduction ; }, abstract = {Genetic pest management (GPM) methods involve releasing modified versions of a pest species to mate with wild pests in the target area. Proposed for a wide range of applications in public health, agriculture and conservation, most progress has been made with pest insects. Offspring of the released modified insects and wild pests carry the modification-which might be transgenes, artificially introduced Wolbachia or genetic damage from radiation, for example-but they also carry a complete haploid genome from their laboratory-reared parent, as well as one from their wild parent. Unless these F1 hybrids are completely unable to reproduce, further mating will lead to introgression of DNA sequences from the release strain into the wild population. We discuss issues around strain selection and the potential consequences of such introgression. We conclude that such introgression is probably harmless in almost all circumstances, and could, in theory, provide specific additional benefits to the release programme. We outline population monitoring approaches that could be used, going forward, to determine how background genetics may affect GPM. This article is part of the theme issue 'Novel control strategies for mosquito-borne diseases'.}, } @article {pmid33357050, year = {2021}, author = {Ahmad, NA and Mancini, MV and Ant, TH and Martinez, J and Kamarul, GMR and Nazni, WA and Hoffmann, AA and Sinkins, SP}, title = {Wolbachia strain wAlbB maintains high density and dengue inhibition following introduction into a field population of Aedes aegypti.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {376}, number = {1818}, pages = {20190809}, pmid = {33357050}, issn = {1471-2970}, support = {202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; 202888/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Aedes ; Animals ; Dengue/*prevention & control ; Dengue Virus/physiology ; Malaysia ; *Mosquito Vectors ; *Pest Control, Biological ; Virus Replication ; Wolbachia/*genetics ; }, abstract = {Aedes aegypti mosquitoes carrying the wAlbB Wolbachia strain show a reduced capacity to transmit dengue virus. wAlbB has been introduced into wild Ae. aegypti populations in several field sites in Kuala Lumpur, Malaysia, where it has persisted at high frequency for more than 2 years and significantly reduced dengue incidence. Although these encouraging results indicate that wAlbB releases can be an effective dengue control strategy, the long-term success depends on wAlbB maintaining high population frequencies and virus transmission inhibition, and both could be compromised by Wolbachia-host coevolution in the field. Here, wAlbB-carrying Ae. aegypti collected from the field 20 months after the cessation of releases showed no reduction in Wolbachia density or tissue distribution changes compared to a wAlbB laboratory colony. The wAlbB strain continued to induce complete unidirectional cytoplasmic incompatibility, showed perfect maternal transmission under laboratory conditions, and retained its capacity to inhibit dengue. Additionally, a field-collected wAlbB line was challenged with Malaysian dengue patient blood, and showed significant blocking of virus dissemination to the salivary glands. These results indicate that wAlbB continues to inhibit currently circulating strains of dengue in field populations of Ae. aegypti, and provides additional support for the continued scale-up of Wolbachia wAlbB releases for dengue control. This article is part of the theme issue 'Novel control strategies for mosquito-borne diseases'.}, } @article {pmid33352133, year = {2020}, author = {Sullivan, W}, title = {Vector Control: Wolbachia Expands Its Protective Reach from Humans to Plants.}, journal = {Current biology : CB}, volume = {30}, number = {24}, pages = {R1489-R1491}, doi = {10.1016/j.cub.2020.11.005}, pmid = {33352133}, issn = {1879-0445}, mesh = {Animals ; *Hemiptera ; Humans ; Insect Vectors ; *Oryza ; Plant Diseases/prevention & control ; *Reoviridae ; *Wolbachia ; }, abstract = {RNA viral titers are often suppressed in insects co-infected with the bacterial endosymbiont Wolbachia. This property has been used to suppress transmission of the ragged rice stunt virus from its insect host, the brown planthopper, to the rice plant.}, } @article {pmid33325576, year = {2021}, author = {Bishop, C and Asgari, S}, title = {Altered gene expression profile of Wolbachia pipientis wAlbB strain following transinfection from its native host Aedes albopictus to Aedes aegypti cells.}, journal = {Molecular microbiology}, volume = {115}, number = {6}, pages = {1229-1243}, doi = {10.1111/mmi.14668}, pmid = {33325576}, issn = {1365-2958}, mesh = {Aedes/*microbiology ; Animals ; Antibiosis ; Bacterial Outer Membrane Proteins/biosynthesis ; Cell Line ; Down-Regulation/genetics ; Gene Expression/genetics ; Gene Expression Regulation, Bacterial/*genetics ; Mitogen-Activated Protein Kinase 3/biosynthesis ; Mosquito Vectors/microbiology/virology ; Polymorphism, Single Nucleotide/genetics ; Sodium-Hydrogen Exchangers/biosynthesis ; Symbiosis/*physiology ; Vector Borne Diseases/prevention & control/virology ; Virus Replication/physiology ; Wolbachia/*genetics/*metabolism ; Yellow Fever/transmission ; Yellow fever virus/growth & development ; }, abstract = {Wolbachia is an obligate intracellular bacterial symbiont prevalent among arthropods and nematodes. To survive and reproduce, Wolbachia interacts with and modifies host subcellular structures, while sensing and responding to changes within the cellular environment. In mutualistic associations, Wolbachia may provision the host with metabolites, or help to maintain the chemical homeostasis of the host cell. Some strains can rapidly invade insect populations by manipulating host reproductive biology, while also preventing viral replication, allowing their use in vector control of arthropod-borne viruses. The Aedes albopictus-derived strain wAlbB is promising in this regard. When transinfected into the Yellow fever mosquito, Aedes aegypti, wAlbB reaches high frequencies within wild populations, and strongly inhibits viral transmission. Despite its obvious potential, much is still unknown about the molecular interactions between Wolbachia and host that enable its use in vector control. Furthermore, most Wolbachia transinfection research to date has focused on host effects. In the current study, we used a cell line model to explore the effect of transinfection of wAlbB from Ae. albopictus to Ae. aegypti. Using RNA sequencing, we show that several genes associated with host-symbiont interactions were downregulated by transinfection, with the greatest downregulation exhibited by prophage-associated genes.}, } @article {pmid33315074, year = {2020}, author = {Cardoso, A and Gómez-Zurita, J}, title = {Food Resource Sharing of Alder Leaf Beetle Specialists (Coleoptera: Chrysomelidae) as Potential Insect-Plant Interface for Horizontal Transmission of Endosymbionts.}, journal = {Environmental entomology}, volume = {49}, number = {6}, pages = {1402-1414}, pmid = {33315074}, issn = {1938-2936}, mesh = {Animals ; *Coleoptera ; Insecta ; North America ; Phylogeny ; *RNA, Ribosomal, 16S/genetics ; Specialization ; Symbiosis ; }, abstract = {Recent studies suggest that endosymbionts of herbivore insects can be horizontally transferred to other herbivores feeding on the same host plants, whereby the plant acts as an intermediate stage in the chain of transmission. If this mechanism operates, it is also expected that insect communities sharing the same host plant will have higher chances to share their endosymbionts. In this study, we use a high-throughput 16S rRNA metabarcoding approach to investigate the presence, diversity, and potential sharing of endosymbionts in several species of leaf beetles (Coleoptera: Chrysomelidae) of a local community specialized on an alder diet in North America. Rickettsia and Wolbachia were predominant in the sample, with strong evidence for each species having their own dominant infection, of either or both types of bacteria. However, all species shared a much lower proportion of a particular Wolbachia type, compatible with the same strain dominant in one of the species of leaf beetles. Crucially, the same 16S rRNA haplotype of Wolbachia was found on alder leaf extracts. The combined evidence and the absence of this strain in a syntopic species of leaf beetle feeding on a different host plant support the hypothesis that at least the initial stages of the mechanism that would allow horizontal transmission of endosymbionts across species feeding on the same plant is possible. The accessibility and characteristics of endosymbiont associations of this system make it suitable for deeper analyses of their diversity and transmission in natural conditions.}, } @article {pmid33298138, year = {2020}, author = {Ding, H and Yeo, H and Puniamoorthy, N}, title = {Wolbachia infection in wild mosquitoes (Diptera: Culicidae): implications for transmission modes and host-endosymbiont associations in Singapore.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {612}, pmid = {33298138}, issn = {1756-3305}, support = {R154000A56133//National University of Singapore/ ; R154000A75114//Ministry of Education, Singapore/ ; }, mesh = {Aedes/genetics/microbiology ; Anaplasmataceae Infections/*microbiology/*transmission ; Animals ; Biological Evolution ; Culex/genetics/microbiology ; Culicidae/*microbiology ; DNA, Bacterial ; Female ; Host-Pathogen Interactions/*physiology ; Male ; Mosquito Vectors/*microbiology ; Polymerase Chain Reaction ; Sequence Analysis ; Singapore ; Vector Borne Diseases ; Wolbachia/classification/genetics ; }, abstract = {BACKGROUND: Wolbachia are intracellular bacterial endosymbionts found in most insect lineages. In mosquitoes, the influence of these endosymbionts on host reproduction and arboviral transmission has spurred numerous studies aimed at using Wolbachia infection as a vector control technique. However, there are several knowledge gaps in the literature and little is known about natural Wolbachia infection across species, their transmission modes, or associations between various Wolbachia lineages and their hosts. This study aims to address these gaps by exploring mosquito-Wolbachia associations and their evolutionary implications.

METHODS: We conducted tissue-specific polymerase chain reaction screening for Wolbachia infection in the leg, gut and reproductive tissues of wild mosquitoes from Singapore using the Wolbachia surface protein gene (wsp) molecular marker. Mosquito-Wolbachia associations were explored using three methods-tanglegram, distance-based, and event-based methods-and by inferred instances of vertical transmission and host shifts.

RESULTS: Adult mosquitoes (271 specimens) representing 14 genera and 40 species were screened for Wolbachia. Overall, 21 species (51.2%) were found positive for Wolbachia, including five in the genus Aedes and five in the genus Culex. To our knowledge, Wolbachia infections have not been previously reported in seven of these 21 species: Aedes nr. fumidus, Aedes annandalei, Uranotaenia obscura, Uranotaenia trilineata, Verrallina butleri, Verrallina sp. and Zeugnomyia gracilis. Wolbachia were predominantly detected in the reproductive tissues, which is an indication of vertical transmission. However, Wolbachia infection rates varied widely within a mosquito host species. There was no clear signal of cophylogeny between the mosquito hosts and the 12 putative Wolbachia strains observed in this study. Host shift events were also observed.

CONCLUSIONS: Our results suggest that the mosquito-Wolbachia relationship is complex and that combinations of transmission modes and multiple evolutionary events likely explain the observed distribution of Wolbachia diversity across mosquito hosts. These findings have implications for a better understanding of the diversity and ecology of Wolbachia and for their utility as biocontrol agents.}, } @article {pmid33295865, year = {2020}, author = {Lefoulon, E and Clark, T and Guerrero, R and Cañizales, I and Cardenas-Callirgos, JM and Junker, K and Vallarino-Lhermitte, N and Makepeace, BL and Darby, AC and Foster, JM and Martin, C and Slatko, BE}, title = {Diminutive, degraded but dissimilar: Wolbachia genomes from filarial nematodes do not conform to a single paradigm.}, journal = {Microbial genomics}, volume = {6}, number = {12}, pages = {}, pmid = {33295865}, issn = {2057-5858}, mesh = {Animals ; Databases, Genetic ; Evolution, Molecular ; Filarioidea/*microbiology ; Genome Size ; Genome, Bacterial ; Genomics ; Molecular Sequence Annotation ; Phylogeny ; Sequence Analysis, DNA/*methods ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {Wolbachia are alpha-proteobacteria symbionts infecting a large range of arthropod species and two different families of nematodes. Interestingly, these endosymbionts are able to induce diverse phenotypes in their hosts: they are reproductive parasites within many arthropods, nutritional mutualists within some insects and obligate mutualists within their filarial nematode hosts. Defining Wolbachia 'species' is controversial and so they are commonly classified into 17 different phylogenetic lineages, termed supergroups, named A-F, H-Q and S. However, available genomic data remain limited and not representative of the full Wolbachia diversity; indeed, of the 24 complete genomes and 55 draft genomes of Wolbachia available to date, 84 % belong to supergroups A and B, exclusively composed of Wolbachia from arthropods. For the current study, we took advantage of a recently developed DNA-enrichment method to produce four complete genomes and two draft genomes of Wolbachia from filarial nematodes. Two complete genomes, wCtub and wDcau, are the smallest Wolbachia genomes sequenced to date (863 988 bp and 863 427 bp, respectively), as well as the first genomes representing supergroup J. These genomes confirm the validity of this supergroup, a controversial clade due to weaknesses of the multilocus sequence typing approach. We also produced the first draft Wolbachia genome from a supergroup F filarial nematode representative (wMhie), two genomes from supergroup D (wLsig and wLbra) and the complete genome of wDimm from supergroup C. Our new data confirm the paradigm of smaller Wolbachia genomes from filarial nematodes containing low levels of transposable elements and the absence of intact bacteriophage sequences, unlike many Wolbachia from arthropods, where both are more abundant. However, we observe differences among the Wolbachia genomes from filarial nematodes: no global co-evolutionary pattern, strong synteny between supergroup C and supergroup J Wolbachia, and more transposable elements observed in supergroup D Wolbachia compared to the other supergroups. Metabolic pathway analysis indicates several highly conserved pathways (haem and nucleotide biosynthesis, for example) as opposed to more variable pathways, such as vitamin B biosynthesis, which might be specific to certain host-symbiont associations. Overall, there appears to be no single Wolbachia-filarial nematode pattern of co-evolution or symbiotic relationship.}, } @article {pmid33294020, year = {2020}, author = {Fu, Z and Meier, AR and Epstein, B and Bergland, AO and Castillo Carrillo, CI and Cooper, WR and Cruzado, RK and Horton, DR and Jensen, AS and Kelley, JL and Rashed, A and Reitz, SR and Rondon, SI and Thinakaran, J and Wenninger, EJ and Wohleb, CH and Crowder, DW and Snyder, WE}, title = {Host plants and Wolbachia shape the population genetics of sympatric herbivore populations.}, journal = {Evolutionary applications}, volume = {13}, number = {10}, pages = {2740-2753}, pmid = {33294020}, issn = {1752-4571}, support = {F32 GM097837/GM/NIGMS NIH HHS/United States ; }, abstract = {Changing climate and land-use practices have the potential to bring previously isolated populations of pest insects into new sympatry. This heightens the need to better understand how differing patterns of host-plant association, and unique endosymbionts, serve to promote genetic isolation or integration. We addressed these factors in populations of potato psyllid, Bactericera cockerelli (Šulc), a generalist herbivore that vectors a bacterial pathogen (Candidatus Liberibacter solanacearum, causal pathogen of zebra chip disease) of potato (Solanum tuberosum L.). Genome-wide SNP data revealed two major genetic clusters-psyllids collected from potato crops were genetically similar to psyllids found on a common weed, Lycium spp., but dissimilar from those found on another common non-crop host, Solanum dulcamara L. Most psyllids found on Lycium spp. and potato represented a single mitochondrial cytochrome oxidase I (COI) haplotype that has been suggested to not be native to the region, and whose arrival may have been concurrent with zebra chip disease first emerging. The putatively introduced COI haplotype usually co-occurred with endosymbiotic Wolbachia, while the putatively resident COI haplotype generally did not. Genetic intermediates between the two genetic populations of insects were rare, consistent with recent sympatry or reproductive isolation, although admixture patterns of apparent hybrids were consistent with introgression of genes from introduced into resident populations. Our results suggest that both host-plant associations and endosymbionts are shaping the population genetic structure of sympatric psyllid populations associated with different non-crop hosts. It is of future interest to explicitly examine vectorial capacity of the two populations and their potential hybrids, as population structure and hybridization might alter regional vector capacity and disease outbreaks.}, } @article {pmid33293886, year = {2020}, author = {Wöger, R and Wöger, R and Nuss, M}, title = {DNA barcodes for Aotearoa New Zealand Pyraloidea (Lepidoptera).}, journal = {Biodiversity data journal}, volume = {8}, number = {}, pages = {e58841}, pmid = {33293886}, issn = {1314-2828}, abstract = {Identification of pyraloid species is often hampered by highly similar external morphology requiring microscopic dissection of genitalia. This becomes especially obvious when mass samples from ecological studies or insect monitoring have to be analysed. DNA barcode sequences could accelerate identification, but are not available for most pyraloid species from New Zealand. Hence, we are presenting a first DNA-barcode library for this group, providing 440 COI barcodes (cytochrome C oxidase I sequences) for 73 morphologically-identified species, which is 29% of Pyraloidea known from New Zealand. Results are analysed using the Barcode Index Number system (BIN) of BOLD and the Automatic Barcode Gap Discovery method (ABGD). Using BIN, the 440 barcodes reveal 82 clusters. A perfect match between BIN assignment and morphological identification was found for 63 species (86.3%). Four species (5.5%) share BINs, each with two species in one BIN, of which Glaucocharis epiphaea and Glaucocharis harmonica even share the same barcode. In contrast, six species (8.2%) split into two or more BINs, with the highest number of five BINs for Orocrambus ramosellus. The interspecific variation of all collected specimens of New Zealand Pyraloidea averages 12.54%. There are deep intraspecific divergences (> 2%) in seven species, for instance Orocrambus vulgaris with up to 6.6% and Scoparia ustimacula with 5.5%. Using ABGD, the 440 barcodes reveal 71 or 88 operational taxonomic units (OTUs), depending on the preferred partition. A perfect match between OTU and morphological identification was found for 56 species (76.7%) or 62 species (84.9%). ABGD delivers four or seven species sharing OTUs and four or ten species split into more than one OTU. Morphological re-examination, as well as the analysis of a concatenated dataset of COI and the nuclear markers EF1α and GADPH for species split into more than one BIN or OTU, do not support a higher number of species. Likewise, there is no evidence for Wolbachia infection as a trigger for these sequence variations.}, } @article {pmid33284808, year = {2020}, author = {Schiefer, A and Hübner, MP and Krome, A and Lämmer, C and Ehrens, A and Aden, T and Koschel, M and Neufeld, H and Chaverra-Muñoz, L and Jansen, R and Kehraus, S and König, GM and Pogorevc, D and Müller, R and Stadler, M and Hüttel, S and Hesterkamp, T and Wagner, K and Pfarr, K and Hoerauf, A}, title = {Corallopyronin A for short-course anti-wolbachial, macrofilaricidal treatment of filarial infections.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {12}, pages = {e0008930}, pmid = {33284808}, issn = {1935-2735}, mesh = {Animals ; Female ; Filariasis/*drug therapy/parasitology ; Filaricides/*therapeutic use ; Filarioidea/*drug effects/microbiology ; Lactones/*therapeutic use ; Mice ; Mice, Inbred BALB C ; Symbiosis/drug effects ; Wolbachia/*drug effects ; }, abstract = {Current efforts to eliminate the neglected tropical diseases onchocerciasis and lymphatic filariasis, caused by the filarial nematodes Onchocerca volvulus and Wuchereria bancrofti or Brugia spp., respectively, are hampered by lack of a short-course macrofilaricidal-adult-worm killing-treatment. Anti-wolbachial antibiotics, e.g. doxycycline, target the essential Wolbachia endosymbionts of filariae and are a safe prototype adult-worm-sterilizing and macrofilaricidal regimen, in contrast to standard treatments with ivermectin or diethylcarbamazine, which mainly target the microfilariae. However, treatment regimens of 4-5 weeks necessary for doxycycline and contraindications limit its use. Therefore, we tested the preclinical anti-Wolbachia drug candidate Corallopyronin A (CorA) for in vivo efficacy during initial and chronic filarial infections in the Litomosoides sigmodontis rodent model. CorA treatment for 14 days beginning immediately after infection cleared >90% of Wolbachia endosymbionts from filariae and prevented development into adult worms. CorA treatment of patently infected microfilaremic gerbils for 14 days with 30 mg/kg twice a day (BID) achieved a sustained reduction of >99% of Wolbachia endosymbionts from adult filariae and microfilariae, followed by complete inhibition of filarial embryogenesis resulting in clearance of microfilariae. Combined treatment of CorA and albendazole, a drug currently co-administered during mass drug administrations and previously shown to enhance efficacy of anti-Wolbachia drugs, achieved microfilarial clearance after 7 days of treatment at a lower BID dose of 10 mg/kg CorA, a Human Equivalent Dose of 1.4 mg/kg. Importantly, this combination led to a significant reduction in the adult worm burden, which has not yet been published with other anti-Wolbachia candidates tested in this model. In summary, CorA is a preclinical candidate for filariasis, which significantly reduces treatment times required to achieve sustained Wolbachia depletion, clearance of microfilariae, and inhibition of embryogenesis. In combination with albendazole, CorA is robustly macrofilaricidal after 7 days of treatment and fulfills the Target Product Profile for a macrofilaricidal drug.}, } @article {pmid33283864, year = {2020}, author = {Wang, X and Xiong, X and Cao, W and Zhang, C and Werren, JH and Wang, X}, title = {Phylogenomic Analysis of Wolbachia Strains Reveals Patterns of Genome Evolution and Recombination.}, journal = {Genome biology and evolution}, volume = {12}, number = {12}, pages = {2508-2520}, pmid = {33283864}, issn = {1759-6653}, mesh = {*Genome, Bacterial ; *Phylogeny ; *Recombination, Genetic ; Wolbachia/*genetics ; }, abstract = {Wolbachia are widespread intracellular bacteria that mediate many important biological processes in arthropod species. In this study, we identified 210 conserved single-copy genes in 33 genome-sequenced Wolbachia strains in the A-F supergroups. Phylogenomic analyses with these core genes indicate that all 33 Wolbachia strains maintain the supergroup relationship, which was classified previously based on the multilocus sequence typing (MLST) genes. Using an interclade recombination screening method, 14 inter-supergroup recombination events were discovered in six genes (2.9%) among 210 single-copy orthologs. This finding suggests a relatively low frequency of intergroup recombination. Interestingly, they have occurred not only between A and B supergroups (nine events) but also between A and E supergroups (five events). Maintenance of such transfers suggests possible roles in Wolbachia infection-related functions. Comparisons of strain divergence using the five genes of the MLST system show a high correlation (Pearson correlation coefficient r = 0.98) between MLST and whole-genome divergences, indicating that MLST is a reliable method for identifying related strains when whole-genome data are not available. The phylogenomic analysis and the identified core gene set in our study will serve as a valuable foundation for strain identification and the investigation of recombination and genome evolution in Wolbachia.}, } @article {pmid33281793, year = {2020}, author = {Miao, YH and Xiao, JH and Huang, DW}, title = {Distribution and Evolution of the Bacteriophage WO and Its Antagonism With Wolbachia.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {595629}, pmid = {33281793}, issn = {1664-302X}, abstract = {The symbiosis system comprising eukaryotic hosts, intracellular bacterium Wolbachia, and temperate bacteriophages WO is widely spread through nearly half the number of arthropod species. The relationships between the three components of the system are extremely intricate. Even though the bacteriophage WO can have diverse influences on the ecology and evolution of Wolbachia, little is known about the distribution and evolution of the phages. To the best of our knowledge, this study is the first to report that in infected fig wasps (Ceratosolen solmsi, Kradibia gibbosae, and Wiebesia pumilae), the genomes of all the Wolbachia strains had only one cryptic WO prophage, which contained defects in the genomic structural modules. This phenomenon was contrary to the widely accepted understanding that Wolbachia with cryptic prophages usually possesses at least one intact WO prophage consisting of gene sequences of the head, baseplate, and tail modules, through which the prophage could form intact virions. In addition to the genetic structure features, the phylogenetic relationships of WO and Wolbachia also revealed that bacteriophage WO can horizontally spread among a certain genus or a group of insect hosts, nearly free from the restriction of the affiliation of Wolbachia. Combined with the vertical transmission along with Wolbachia, the wide spread of WO phages can be explained. Furthermore, the gender preference and functional module preference for transcriptional activity of the genes in cryptic WOs implied the antagonized coevolutionary pattern between WO prophages and their Wolbachia hosts.}, } @article {pmid33276723, year = {2020}, author = {Yang, C and Zheng, Y and Tan, S and Meng, G and Rao, W and Yang, C and Bourne, DG and O'Brien, PA and Xu, J and Liao, S and Chen, A and Chen, X and Jia, X and Zhang, AB and Liu, S}, title = {Efficient COI barcoding using high throughput single-end 400 bp sequencing.}, journal = {BMC genomics}, volume = {21}, number = {1}, pages = {862}, pmid = {33276723}, issn = {1471-2164}, support = {NO. JCYJ20170817150755701//Shenzhen Municipal Government of China/ ; No. KQTD20150330171505310//Shenzhen Peacock Plan/ ; 31425023//China National Funds for Distinguished Young Scientists/ ; 2019M660051//Chinese Postdoctoral Science Foundation/ ; }, mesh = {Animals ; DNA ; *DNA Barcoding, Taxonomic ; *Ecosystem ; High-Throughput Nucleotide Sequencing ; Insecta ; }, abstract = {BACKGROUND: Over the last decade, the rapid development of high-throughput sequencing platforms has accelerated species description and assisted morphological classification through DNA barcoding. However, the current high-throughput DNA barcoding methods cannot obtain full-length barcode sequences due to read length limitations (e.g. a maximum read length of 300 bp for the Illumina's MiSeq system), or are hindered by a relatively high cost or low sequencing output (e.g. a maximum number of eight million reads per cell for the PacBio's SEQUEL II system).

RESULTS: Pooled cytochrome c oxidase subunit I (COI) barcodes from individual specimens were sequenced on the MGISEQ-2000 platform using the single-end 400 bp (SE400) module. We present a bioinformatic pipeline, HIFI-SE, that takes reads generated from the 5' and 3' ends of the COI barcode region and assembles them into full-length barcodes. HIFI-SE is written in Python and includes four function modules of filter, assign, assembly and taxonomy. We applied the HIFI-SE to a set of 845 samples (30 marine invertebrates, 815 insects) and delivered a total of 747 fully assembled COI barcodes as well as 70 Wolbachia and fungi symbionts. Compared to their corresponding Sanger sequences (72 sequences available), nearly all samples (71/72) were correctly and accurately assembled, including 46 samples that had a similarity score of 100% and 25 of ca. 99%.

CONCLUSIONS: The HIFI-SE pipeline represents an efficient way to produce standard full-length barcodes, while the reasonable cost and high sensitivity of our method can contribute considerably more DNA barcodes under the same budget. Our method thereby advances DNA-based species identification from diverse ecosystems and increases the number of relevant applications.}, } @article {pmid33263885, year = {2020}, author = {Attardo, GM and Scolari, F and Malacrida, A}, title = {Bacterial Symbionts of Tsetse Flies: Relationships and Functional Interactions Between Tsetse Flies and Their Symbionts.}, journal = {Results and problems in cell differentiation}, volume = {69}, number = {}, pages = {497-536}, pmid = {33263885}, issn = {0080-1844}, mesh = {Animals ; Enterobacteriaceae/*physiology ; Female ; *Host Microbial Interactions ; Male ; Microbiota ; *Symbiosis ; Trypanosoma ; Tsetse Flies/*microbiology/parasitology ; Wolbachia/*physiology ; }, abstract = {Tsetse flies (Glossina spp.) act as the sole vectors of the African trypanosome species that cause Human African Trypanosomiasis (HAT or African Sleeping Sickness) and Nagana in animals. These flies have undergone a variety of specializations during their evolution including an exclusive diet consisting solely of vertebrate blood for both sexes as well as an obligate viviparous reproductive biology. Alongside these adaptations, Glossina species have developed intricate relationships with specific microbes ranging from mutualistic to parasitic. These relationships provide fundamental support required to sustain the specializations associated with tsetse's biology. This chapter provides an overview on the knowledge to date regarding the biology behind these relationships and focuses primarily on four bacterial species that are consistently associated with Glossina species. Here their interactions with the host are reviewed at the morphological, biochemical and genetic levels. This includes: the obligate symbiont Wigglesworthia, which is found in all tsetse species and is essential for nutritional supplementation to the blood-specific diet, immune system maturation and facilitation of viviparous reproduction; the commensal symbiont Sodalis, which is a frequently associated symbiont optimized for survival within the fly via nutritional adaptation, vertical transmission through mating and may alter vectorial capacity of Glossina for trypanosomes; the parasitic symbiont Wolbachia, which can manipulate Glossina via cytoplasmic incompatibility and shows unique interactions at the genetic level via horizontal transmission of its genetic material into the genome in two Glossina species; finally, knowledge on recently observed relations between Spiroplasma and Glossina is explored and potential interactions are discussed based on knowledge of interactions between this bacterial Genera and other insect species. These flies have a simple microbiome relative to that of other insects. However, these relationships are deep, well-studied and provide a window into the complexity and function of host/symbiont interactions in an important disease vector.}, } @article {pmid33263882, year = {2020}, author = {Lefoulon, E and Foster, JM and Truchon, A and Carlow, CKS and Slatko, BE}, title = {The Wolbachia Symbiont: Here, There and Everywhere.}, journal = {Results and problems in cell differentiation}, volume = {69}, number = {}, pages = {423-451}, pmid = {33263882}, issn = {0080-1844}, mesh = {Animals ; Arthropods/*microbiology ; Filarioidea/*microbiology ; Host Microbial Interactions ; *Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia symbionts, first observed in the 1920s, are now known to be present in about 30-70% of tested arthropod species, in about half of tested filarial nematodes (including the majority of human filarial nematodes), and some plant-parasitic nematodes. In arthropods, they are generally viewed as parasites while in nematodes they appear to be mutualists although this demarcation is not absolute. Their presence in arthropods generally leads to reproductive anomalies, while in nematodes, they are generally required for worm development and reproduction. In mosquitos, Wolbachia inhibit RNA viral infections, leading to populational reductions in human RNA virus pathogens, whereas in filarial nematodes, their requirement for worm fertility and survival has been channeled into their use as drug targets for filariasis control. While much more research on these ubiquitous symbionts is needed, they are viewed as playing significant roles in biological processes, ranging from arthropod speciation to human health.}, } @article {pmid33263871, year = {2020}, author = {Russell, SL and Castillo, JR}, title = {Trends in Symbiont-Induced Host Cellular Differentiation.}, journal = {Results and problems in cell differentiation}, volume = {69}, number = {}, pages = {137-176}, pmid = {33263871}, issn = {0080-1844}, support = {K99 GM135583/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology ; *Cell Differentiation ; Genome, Bacterial ; Nematoda/*microbiology ; Reproduction ; *Symbiosis ; *Wolbachia/genetics ; }, abstract = {Bacteria participate in a wide diversity of symbiotic associations with eukaryotic hosts that require precise interactions for bacterial recognition and persistence. Most commonly, host-associated bacteria interfere with host gene expression to modulate the immune response to the infection. However, many of these bacteria also interfere with host cellular differentiation pathways to create a hospitable niche, resulting in the formation of novel cell types, tissues, and organs. In both of these situations, bacterial symbionts must interact with eukaryotic regulatory pathways. Here, we detail what is known about how bacterial symbionts, from pathogens to mutualists, control host cellular differentiation across the central dogma, from epigenetic chromatin modifications, to transcription and mRNA processing, to translation and protein modifications. We identify four main trends from this survey. First, mechanisms for controlling host gene expression appear to evolve from symbionts co-opting cross-talk between host signaling pathways. Second, symbiont regulatory capacity is constrained by the processes that drive reductive genome evolution in host-associated bacteria. Third, the regulatory mechanisms symbionts exhibit correlate with the cost/benefit nature of the association. And, fourth, symbiont mechanisms for interacting with host genetic regulatory elements are not bound by native bacterial capabilities. Using this knowledge, we explore how the ubiquitous intracellular Wolbachia symbiont of arthropods and nematodes may modulate host cellular differentiation to manipulate host reproduction. Our survey of the literature on how infection alters gene expression in Wolbachia and its hosts revealed that, despite their intermediate-sized genomes, different strains appear capable of a wide diversity of regulatory manipulations. Given this and Wolbachia's diversity of phenotypes and eukaryotic-like proteins, we expect that many symbiont-induced host differentiation mechanisms will be discovered in this system.}, } @article {pmid33257667, year = {2020}, author = {Souto-Maior, C and King, JG and Sartori, LM and Maciel-de-Freitas, R and Gomes, MGM}, title = {Reply to: "Enhancement of Aedes aegypti susceptibility to dengue by Wolbachia is not supported".}, journal = {Nature communications}, volume = {11}, number = {1}, pages = {6113}, pmid = {33257667}, issn = {2041-1723}, mesh = {*Aedes ; Animals ; *Dengue ; *Wolbachia ; }, } @article {pmid33257651, year = {2020}, author = {Ant, TH and Mancini, MV and Martinez, J and Sinkins, SP}, title = {Enhancement of Aedes aegypti susceptibility to dengue by Wolbachia is not supported.}, journal = {Nature communications}, volume = {11}, number = {1}, pages = {6111}, pmid = {33257651}, issn = {2041-1723}, support = {202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Aedes ; Animals ; *Dengue ; *Wolbachia ; }, } @article {pmid33248417, year = {2020}, author = {Madhav, M and Baker, D and Morgan, JAT and Asgari, S and James, P}, title = {Wolbachia: A tool for livestock ectoparasite control.}, journal = {Veterinary parasitology}, volume = {288}, number = {}, pages = {109297}, doi = {10.1016/j.vetpar.2020.109297}, pmid = {33248417}, issn = {1873-2550}, mesh = {Animals ; Camelus/parasitology ; Cattle/parasitology ; Cimicidae/microbiology ; Diptera/microbiology ; Ectoparasitic Infestations/parasitology/prevention & control/*veterinary ; Goats/parasitology ; Horses/parasitology ; *Livestock ; Mites/microbiology ; Phthiraptera/microbiology ; Sheep, Domestic/parasitology ; Siphonaptera/microbiology ; Sus scrofa/parasitology ; Ticks/microbiology ; Wolbachia/*physiology ; }, abstract = {Ectoparasites and livestock-associated insects are a major concern throughout the world because of their economic and welfare impacts. Effective control is challenging and relies mainly on the use of chemical insecticides and acaricides. Wolbachia, an arthropod and nematode-infecting, maternally-transmitted endosymbiont is currently of widespread interest for use in novel strategies for the control of a range of arthropod-vectored human diseases and plant pests but to date has received only limited consideration for use in the control of diseases of veterinary concern. Here, we review the currently available information on Wolbachia in veterinary ectoparasites and disease vectors, consider the feasibility for use of Wolbachia in the control of livestock pests and diseases and highlight critical issues which need further investigation.}, } @article {pmid33240234, year = {2020}, author = {Momtaz, AZ and Ahumada Sabagh, AD and Gonzalez Amortegui, JG and Salazar, SA and Finessi, A and Hernandez, J and Christensen, S and Serbus, LR}, title = {A Role for Maternal Factors in Suppressing Cytoplasmic Incompatibility.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {576844}, pmid = {33240234}, issn = {1664-302X}, abstract = {Wolbachia are maternally transmitted bacterial endosymbionts, carried by approximately half of all insect species. Wolbachia prevalence in nature stems from manipulation of host reproduction to favor the success of infected females. The best known reproductive modification induced by Wolbachia is referred to as sperm-egg Cytoplasmic Incompatibility (CI). In CI, the sperm of Wolbachia-infected males cause embryonic lethality, attributed to paternal chromatin segregation defects during early mitotic divisions. Remarkably, the embryos of Wolbachia-infected females "rescue" CI lethality, yielding egg hatch rates equivalent to uninfected female crosses. Several models have been discussed as the basis for Rescue, and functional evidence indicates a major contribution by Wolbachia CI factors. A role for host contributions to Rescue remains largely untested. In this study, we used a chemical feeding approach to test for CI suppression capabilities by Drosophila simulans. We found that uninfected females exhibited significantly higher CI egg hatch rates in response to seven chemical treatments that affect DNA integrity, cell cycle control, and protein turnover. Three of these treatments suppressed CI induced by endogenous wRi Wolbachia, as well as an ectopic wMel Wolbachia infection. The results implicate DNA integrity as a focal aspect of CI suppression for different Wolbachia strains. The framework presented here, applied to diverse CI models, will further enrich our understanding of host reproductive manipulation by insect endosymbionts.}, } @article {pmid33237151, year = {2020}, author = {Pimentel, AC and Beraldo, CS and Cogni, R}, title = {Host-shift as the cause of emerging infectious diseases: Experimental approaches using Drosophila-virus interactions.}, journal = {Genetics and molecular biology}, volume = {44}, number = {1 Suppl 1}, pages = {e20200197}, pmid = {33237151}, issn = {1415-4757}, abstract = {Host shifts, when a cross-species transmission of a pathogen can lead to successful infections, are the main cause of emerging infectious diseases, such as COVID-19. A complex challenge faced by the scientific community is to address the factors that determine whether the cross-species transmissions will result in spillover or sustained onwards infections. Here we review recent literature and present a perspective on current approaches we are using to understand the mechanisms underlying host shifts. We highlight the usefulness of the interactions between Drosophila species and viruses as an ideal study model. Additionally, we discuss how cross-infection experiments - when pathogens from a natural reservoir are intentionally injected in novel host species- can test the effect cross-species transmissions may have on the fitness of virus and host, and how the host phylogeny may influence this response. We also discuss experiments evaluating how cooccurrence with other viruses or the presence of the endosymbiont bacteria Wolbachia may affect the performance of new viruses in a novel host. Finally, we discuss the need of surveys of virus diversity in natural populations using next-generation sequencing technologies. In the long term, these approaches can contribute to a better understanding of the basic biology of host shifts.}, } @article {pmid33222623, year = {2022}, author = {Amala, M and Richard, M and Saritha, P and Prabhu, D and Veerapandiyan, M and Surekha, K and Jeyakanthan, J}, title = {Molecular evolution, binding site interpretation and functional divergence of aspartate semialdehyde dehydrogenase.}, journal = {Journal of biomolecular structure & dynamics}, volume = {40}, number = {7}, pages = {3223-3241}, doi = {10.1080/07391102.2020.1846619}, pmid = {33222623}, issn = {1538-0254}, mesh = {Amino Acid Sequence ; *Aspartate-Semialdehyde Dehydrogenase/chemistry/genetics ; Binding Sites ; *Evolution, Molecular ; Phylogeny ; }, abstract = {Aspartate Semialdehyde Dehydrogenase (ASDH) is an important enzyme essential for the viability of pathogenic microorganisms. ASDH is mainly involved in amino acid and cell wall biosynthesis of microorganisms, hence it is considered to be a promising target for drug design. This enzyme depicts similar mechanistic function in all microorganisms; although, the kinetic efficiency of an enzyme differs according to their active site residual composition. Therefore, understanding the residual variation and kinetic efficiency of the enzyme would pave new insights in structure-based drug discovery and a novel drug molecule against ASDH. Here, ASDH from Wolbachia endosymbiont of Brugia malayi is used as a prime enzyme to execute evolutionary studies. The phylogenetic analysis was opted to classify 400 sequences of ASDH enzymes based on their structure and electrostatic surfaces. Analysis resulted in 37 monophyletic clades of diverse pathogenic and non-pathogenic organisms. The representative structures of 37 ASDHs from different clades were further deciphered to structural homologues. These enzymes exhibited presence of more positively charged surfaces than negatively charged surfaces in the active site pocket which restrains evolutionary significance. Docking studies of NADP[+] with 37 enzymes reveals that site-specific residual variation in the active site pocket modulates the binding affinity (ranges of -13 to -9 kcal/mol). Type-I and Type-II divergence studies show, no significant functional divergence among ASDH, but residual changes were found among the enzyme that modulates the biochemical characteristics and catalytic efficiency. The present study not only explores residual alteration and catalytic variability, it also aids in the design of species-specific inhibitors.Communicated by Ramaswamy H. Sarma.}, } @article {pmid33216144, year = {2021}, author = {Cormier, A and Chebbi, MA and Giraud, I and Wattier, R and Teixeira, M and Gilbert, C and Rigaud, T and Cordaux, R}, title = {Comparative Genomics of Strictly Vertically Transmitted, Feminizing Microsporidia Endosymbionts of Amphipod Crustaceans.}, journal = {Genome biology and evolution}, volume = {13}, number = {1}, pages = {}, pmid = {33216144}, issn = {1759-6653}, mesh = {Amphipoda/*genetics ; Animals ; Female ; Feminization/*genetics/parasitology ; *Genomics ; Host-Parasite Interactions ; Male ; Microsporidia/*genetics ; Nosema ; Phylogeny ; Wolbachia/genetics ; }, abstract = {Microsporidia are obligate intracellular eukaryotic parasites of vertebrates and invertebrates. Microsporidia are usually pathogenic and undergo horizontal transmission or a mix of horizontal and vertical transmission. However, cases of nonpathogenic microsporidia, strictly vertically transmitted from mother to offspring, have been reported in amphipod crustaceans. Some of them further evolved the ability to feminize their nontransmitting male hosts into transmitting females. However, our understanding of the evolution of feminization in microsporidia is hindered by a lack of genomic resources. We report the sequencing and analysis of three strictly vertically transmitted microsporidia species for which feminization induction has been demonstrated (Nosema granulosis) or is strongly suspected (Dictyocoela muelleri and Dictyocoela roeselum), along with a draft genome assembly of their host Gammarus roeselii. Contrary to horizontally transmitted microsporidia that form environmental spores that can be purified, feminizing microsporidia cannot be easily isolated from their host cells. Therefore, we cosequenced symbiont and host genomic DNA and devised a computational strategy to obtain genome assemblies for the different partners. Genomic comparison with feminizing Wolbachia bacterial endosymbionts of isopod crustaceans indicated independent evolution of feminization in microsporidia and Wolbachia at the molecular genetic level. Feminization thus represents a remarkable evolutionary convergence of eukaryotic and prokaryotic microorganisms. Furthermore, a comparative genomics analysis of microsporidia allowed us to identify several candidate genes for feminization, involving functions such as DNA binding and membrane fusion. The genomic resources we generated contribute to establish Gammarus roeselii and its microsporidia symbionts as a new model to study the evolution of symbiont-mediated feminization.}, } @article {pmid33198811, year = {2020}, author = {Tongkrajang, N and Ruenchit, P and Tananchai, C and Chareonviriyaphap, T and Kulkeaw, K}, title = {Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {579}, pmid = {33198811}, issn = {1756-3305}, support = {RSA6280102//Research Career Development Grant from Thailand Science Research and Innovation/ ; R016233004//Faculty of Medicine Siriraj Hospital, Mahidol University (TH)/ ; }, mesh = {Animals ; Anopheles/*microbiology ; Female ; Malaria/prevention & control ; Mosquito Vectors/*microbiology ; Myanmar/epidemiology ; Phylogeny ; Thailand/epidemiology ; *Wolbachia/classification/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia, obligate intracellular bacteria, infect the majority of arthropods, including many mosquito species of medical importance. Some Wolbachia strains interfere with the development of Plasmodium parasites in female Anopheles, a major vector of malaria. The use of Wolbachia as a means to block malaria transmission is an emerging vector control strategy in highly endemic areas. Hence, identification of native Wolbachia strains in areas where malaria transmission is low may uncover a particular Wolbachia strain capable of Plasmodium interference. This study aims to identify native Wolbachia strains in female Anopheles spp. that are predominant in a low-malaria transmission area in mainland Southeast Asia.

METHODS: Following a 2-year survey of malaria vectors in Umphang Valley of Tak Province, Thailand, DNA extracts of female An. minimus, An. peditaeniatus, and An. maculatus were subjected to amplification of the conserved region of the 16S rRNA-encoding gene. The DNA sequences of the amplicons were phylogenetically compared with those of known Wolbachia strains.

RESULTS: Among three Anopheles spp., amplification was detected in only the DNA samples from An. minimus. The DNA sequencing of amplicons revealed 100% similarity to Wolbachia pipientis, confirming the specificity of amplification. The Wolbachia-positive An. minimus samples were devoid of Plasmodium 18S rRNA amplification. The phylogenetic trees indicate a close relationship with Wolbachia strains in subgroup B.

CONCLUSION: To the best of our knowledge, the data presented herein provide the first molecular evidence of a Wolbachia strain in An. minimus, hereinafter named wAnmi, in a low-malaria transmission area in the Umphang Valley of western Thailand. Further biological characterization is required to examine its potential for malaria transmission control in the field.}, } @article {pmid33196908, year = {2021}, author = {Chao, LL and Castillo, CT and Shih, CM}, title = {Molecular detection and genetic identification of Wolbachia endosymbiont in Rhipicephalus sanguineus (Acari: Ixodidae) ticks of Taiwan.}, journal = {Experimental & applied acarology}, volume = {83}, number = {1}, pages = {115-130}, pmid = {33196908}, issn = {1572-9702}, support = {MOST 108-2314-B-037-063//Ministry of Science and Technology, Taiwan/ ; }, mesh = {Animals ; Female ; *Ixodidae ; Male ; Phylogeny ; *Rhipicephalus sanguineus ; Taiwan ; *Wolbachia/genetics ; }, abstract = {The genetic identity of Wolbachia endosymbiont in Rhipicephalus sanguineus ticks was determined for the first time in Taiwan. In total 1004 Rh. sanguineus ticks were examined for Wolbachia by polymerase chain reaction assay targeting the Wolbachia surface protein (wsp) gene. The prevalence of Wolbachia infection was detected in nymphs, females, and males with an infection rate of 55.8, 39.8, and 44%, respectively. The phylogenetic relationships were analyzed by comparing the sequences of wsp gene obtained from 60 strains of Wolbachia representing 11 strains of supergroup A and 10 strains of supergroup B. In general, seven major clades of supergroup A and six major clades of supergroup B can be easily distinguished by neighbour-joining analysis and were congruent by maximum likelihood method. All these Wolbachia strains of Taiwan were genetically affiliated to supergroups A and B with high sequence similarity of 98.3-100% and 98.6-100%, respectively. Intra- and inter-group analysis based on the genetic distance (GD) values indicated a lower level (GD < 0.017) within the group A strains of Taiwan compared with the group B (GD > 0.576) of other Wolbachia strains, as well as a lower level (GD < 0.062) within the group B strains of Taiwan compared with the group A (GD > 0.246) of other Wolbachia strains. Our results provide the first genetic identification of Wolbachia endosymbiont in Rh. sanguineus ticks collected from Taiwan, and detection of Wolbachia in male and nymphal ticks may imply the possible mechanism of transstadial transmission in Rh. sanguineus ticks.}, } @article {pmid33193166, year = {2020}, author = {Salgueiro, J and Pimper, LE and Segura, DF and Milla, FH and Russo, RM and Asimakis, E and Stathopoulou, P and Bourtzis, K and Cladera, JL and Tsiamis, G and Lanzavecchia, SB}, title = {Gut Bacteriome Analysis of Anastrepha fraterculus sp. 1 During the Early Steps of Laboratory Colonization.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {570960}, pmid = {33193166}, issn = {1664-302X}, abstract = {Microbial communities associated to insect species are involved in essential biological functions such as host nutrition, reproduction and survivability. Main factors have been described as modulators of gut bacterial community, such as diet, habit, developmental stage and taxonomy of the host. The present work focuses on the complex changes that gut microbial communities go through when wild insects are introduced to artificial rearing conditions. Specifically, we analyzed the effect of the laboratory colonization on the richness and diversity of the gut bacteriome hosted by the fruit fly pest Anastrepha fraterculus sp. 1. Bacterial profiles were studied by amplicon sequencing of the 16S rRNA V3-V4 hypervariable region in gut samples of males and females, in teneral (1-day-old, unfed) and post-teneral (15-day-old, fed) flies. A total of 3,147,665 sequence reads were obtained and 32 bacterial operational taxonomic units (OTUs) were identified. Proteobacteria was the most abundant phylum (93.3% of the total reads) and, Wolbachia and Enterobacter were the most represented taxa at the genus level (29.9% and 27.7%, respectively, of the total read counts). Wild and laboratory flies showed highly significant differences in the relative abundances of bacteria. The analysis of the core bacteriome showed the presence of five OTUs in all samples grouped by origin, while nine and five OTUs were exclusively detected in laboratory and wild flies, respectively. Irrespective of fly origin or sex, a dominant presence of Wolbachia was observed in teneral flies, whereas Enterobacter was highly abundant in post-teneral individuals. We evidenced significant differences in bacterial richness and diversity among generations under laboratory colonization (F0, F1, F3 and F6) and compared to laboratory and wild flies, displaying also differential patterns between teneral and post-teneral flies. Laboratory and wild A. fraterculus sp. 1 harbor different gut bacterial communities. Laboratory colonization has an important effect on the microbiota, most likely associated to the combined effects of insect physiology and environmental conditions (e.g., diet and colony management).}, } @article {pmid33185065, year = {2020}, author = {Wang, YF and He, K and Guo, W and Wang, HF and Zhang, DH and Gong, MQ and Ji, MJ and Chen, L}, title = {[Genes associated with Wolbachia-induced cytoplasmic incompatibility in natural populations of Culex pipiens pallens: a preliminary study].}, journal = {Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control}, volume = {32}, number = {5}, pages = {517-521}, doi = {10.16250/j.32.1374.2019197}, pmid = {33185065}, issn = {1005-6661}, mesh = {Animals ; China ; Culex/*genetics/microbiology ; Cytoplasm/genetics/*microbiology ; *Genes, Insect ; Polymerase Chain Reaction ; *Wolbachia ; }, abstract = {OBJECTIVE: To investigate the genes involved in Wolbachia-induced cytoplasmic incompatibility among three natural populations of Culex pipiens pallens in eastern China, so as to provide insights into the development of preventive and control measures for mosquito-borne diseases based on Wolbachia.

METHODS: The cytoplasmic incompatibility was tested among three natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province using reciprocal crosses. Wolbachia infection was detected in C. pipiens pallens using a PCR assay, and the expression of Wolbachia wsp and WD0513 genes was quantified using a fluorescent quantitative real-time PCR (qPCR) assay.

RESULTS: Bidirectional compatibility was found between the natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province (t = 0.57 and 0.15, both P values > 0.05), while bidirectional incompatibility was seen between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Wuxi of Jiangsu Province (t = 63.81 and 43.51, both P values < 0.01), and between the natural populations of C. pipiens pallens collected from Nanjing of Jiangsu Province and Tangkou of Shandong Province (t = 39.62 and 43.12, both P values < 0.01). Wolbachia wsp gene was amplified in all three natural populations of C. pipiens pallens, and qPCR assay detected no significant difference in the Wolbachia wsp gene expression among the three natural populations of C. pipiens pallens (F = 2.15, P > 0.05). In addition, there was no significant difference in the WD0513 gene expression between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Nanjing of Jiangsu Province (q = 8.42, P < 0.05) or between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Wuxi of Jiangsu Province (q = 7.84, P < 0.05); however, there was a significant difference detected in the WD0513 gene expression between the natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province (q = 0.40, P > 0.05).

CONCLUSIONS: Different Wolbachia numbers are detected in natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province, and WD0513 gene may be involved in the Wolbachia-induced cytoplasmic incompatibility among three natural populations of C. pipiens pallens.}, } @article {pmid33180772, year = {2020}, author = {Arias-Giraldo, LM and Muñoz, M and Hernández, C and Herrera, G and Velásquez-Ortiz, N and Cantillo-Barraza, O and Urbano, P and Ramírez, JD}, title = {Species-dependent variation of the gut bacterial communities across Trypanosoma cruzi insect vectors.}, journal = {PloS one}, volume = {15}, number = {11}, pages = {e0240916}, pmid = {33180772}, issn = {1932-6203}, mesh = {Animals ; Biodiversity ; Chagas Disease/transmission ; Colombia ; DNA, Bacterial/genetics ; DNA, Protozoan/genetics ; *Gastrointestinal Microbiome/genetics ; Genotype ; Host Specificity ; Humans ; Insect Vectors/classification/*microbiology/*parasitology ; RNA, Ribosomal, 16S/genetics ; Triatominae/classification/*microbiology/*parasitology ; Trypanosoma cruzi/classification/genetics/*pathogenicity ; }, abstract = {Triatomines (Hemiptera: Reduviidae) are the insect vectors of Trypanosoma cruzi, the causative agent of Chagas disease. The gut bacterial communities affect the development of T. cruzi inside the vector, making the characterization of its composition important in the understanding of infection development. We collected 54 triatomine bugs corresponding to four genera in different departments of Colombia. DNA extraction and PCR were performed to evaluate T. cruzi presence and to determine the discrete typing unit (DTU) of the parasite. PCR products of the bacterial 16S rRNA gene were pooled and sequenced. Resulting reads were denoised and QIIME 2 was used for the identification of amplicon sequence variants (ASVs). Diversity (alpha and beta diversity) and richness analyses, Circos plots, and principal component analysis (PCA) were also performed. The overall T. cruzi infection frequency was 75.9%, with TcI being the predominant DTU. Approximately 500,000 sequences were analyzed and 27 bacterial phyla were identified. The most abundant phyla were Proteobacteria (33.9%), Actinobacteria (32.4%), Firmicutes (19.6%), and Bacteroidetes (7.6%), which together accounted for over 90% of the gut communities identified in this study. Genera were identified for these main bacterial phyla, revealing the presence of important bacteria such as Rhodococcus, Serratia, and Wolbachia. The composition of bacterial phyla in the gut of the insects was significantly different between triatomine species, whereas no significant difference was seen between the state of T. cruzi infection. We suggest further investigation with the evaluation of additional variables and a larger sample size. To our knowledge, this study is the first characterization of the gut bacterial structure of the main triatomine genera in Colombia.}, } @article {pmid33170730, year = {2021}, author = {Onah, IE and Sumner, S}, title = {DNA barcodes and new primers for nature's pest controllers: the social wasps.}, journal = {Genome}, volume = {64}, number = {5}, pages = {581-590}, doi = {10.1139/gen-2019-0193}, pmid = {33170730}, issn = {1480-3321}, mesh = {Animals ; Biodiversity ; DNA/analysis ; DNA Barcoding, Taxonomic/*methods ; *DNA Primers ; Insecta/genetics ; *Pest Control ; Phylogeny ; Polymerase Chain Reaction ; Wasps/classification/*genetics ; }, abstract = {Globally, biodiversity is declining because of anthropogenic pressures, and this could lead to extinction of some species before they are discovered. The loss of insect taxa is of prime concern, given recent reports of significant declines in the populations of many taxa across the globe. Efforts to document biodiversity have met with several challenges, amongst which are the difficulties in using morphological features to discriminate species, especially in insects. DNA barcoding is a rapid and reliable method for species identification and discovery but choosing appropriate primers to amplify the barcode region without co-amplifying contaminants remains a key challenge. We developed and tested a set of primers for PCR amplification of the DNA barcode region of the COI gene in polistine wasps. We tested their efficacy in 36 species of vespid wasps, and the solitary wasp Zethus miniatus Saussure. Samples were obtained from Africa, Americas, Asia, and Europe. The polistine-specific primers successfully amplified the barcode region for all polistines tested, without amplifying any Wolbachia present; they also worked with many species from the other Vespidae wasp subfamilies. The new primers are valuable for the discovery and accurate documentation of polistine wasps in the four continents.}, } @article {pmid33168082, year = {2020}, author = {Torres, R and Hernandez, E and Flores, V and Ramirez, JL and Joyce, AL}, title = {Wolbachia in mosquitoes from the Central Valley of California, USA.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {558}, pmid = {33168082}, issn = {1756-3305}, mesh = {Aedes/microbiology ; Animals ; Anopheles/microbiology ; Bacterial Typing Techniques ; California ; Culex/microbiology ; Culicidae/*microbiology ; DNA, Bacterial/genetics ; Female ; Mosquito Vectors/microbiology ; Multilocus Sequence Typing ; *Phylogeny ; RNA, Ribosomal, 16S ; Wolbachia/*classification/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia bacteria are widely distributed throughout terrestrial arthropod species. These bacteria can manipulate reproduction and influence the vector competence of their hosts. Recently, Wolbachia have been integrated into vector control programmes for mosquito management. A number of supergroups and strains exist for Wolbachia, and they have yet to be characterized for many mosquito species. In this study, we examined Wolbachia prevalence and their phylogenetic relationship to other Wolbachia, using mosquitoes collected in Merced County in the Central Valley of California.

METHODS: Adult mosquitoes were collected from 85 sites in Merced County, California in 2017 and 2018. Traditional and quantitative PCR were used to investigate the presence or absence and the density of Wolbachia, using Wolbachia-specific 16S rRNA and Wolbachia-surface protein (wsp) genes. The supergroup of Wolbachia was determined, and Multilocus Sequence Typing (MLST) by sequencing five housekeeping genes (coxA, gatB, ftsZ, hcpA and fbpA) was also used to determine Wolbachia supergroup as well as strain.

RESULTS: Over 7100 mosquitoes of 12 species were collected: Aedes melanimon, Ae. nigromaculis, Ae. vexans, Ae. aegypti, Culex pipiens, Cx. stigmatosoma, Cx. tarsalis, Anopheles franciscanus, An. freeborni, An. punctipennis, Culiseta incidens and Cs. inornata. Eight showed evidence of Wolbachia. To our knowledge, this study is the first to report detection of Wolbachia in five of these species (Ae. melanimon, Cx. stigmatosoma, Cx. tarsalis, Cs. incidens and Cs. inornata). Culex pipiens and Cx. stigmatosoma had a high frequency and density of Wolbachia infection, which grouped into supergroup B; Cs. inornata clustered with supergroup A. MLST comparisons identified Cx. pipiens and Cx. stigmatosoma as wPip strain type 9 supergroup B. Six species had moderate to low (< 14%) frequencies of Wolbachia. Four species were negative, Ae. nigromaculis, An. franciscanus, An. freeborni and Ae. aegypti.

CONCLUSIONS: New records of Wolbachia detection were found in mosquitoes from Merced County, California. Culex stigmatosoma and Cs. inornata were new records for Wolbachia supergroup B and A, respectively. Other species with Wolbachia occurred with low frequency and low density. Detection of Wolbachia in mosquitoes can be used to inform potential vector control applications. Future study of Wolbachia within Cx. stigmatosoma and Cs. inornata in California and through the range of these species could further explore Wolbachia infection in these two species.}, } @article {pmid33160070, year = {2020}, author = {Varotto-Boccazzi, I and Epis, S and Arnoldi, I and Corbett, Y and Gabrieli, P and Paroni, M and Nodari, R and Basilico, N and Sacchi, L and Gramiccia, M and Gradoni, L and Tranquillo, V and Bandi, C}, title = {Boosting immunity to treat parasitic infections: Asaia bacteria expressing a protein from Wolbachia determine M1 macrophage activation and killing of Leishmania protozoans.}, journal = {Pharmacological research}, volume = {161}, number = {}, pages = {105288}, doi = {10.1016/j.phrs.2020.105288}, pmid = {33160070}, issn = {1096-1186}, mesh = {Acetobacteraceae/genetics/*immunology/metabolism ; Animals ; Bacterial Outer Membrane Proteins/genetics/*immunology/metabolism ; Cell Line ; Cytokines/metabolism ; Genetic Vectors ; Host-Parasite Interactions ; *Immunity, Innate ; Leishmania infantum/growth & development/*immunology/ultrastructure ; Leishmaniasis Vaccines/genetics/*immunology/metabolism ; *Macrophage Activation ; Macrophages/immunology/metabolism/*microbiology/*parasitology ; Mice ; Nitric Oxide/metabolism ; Phagocytosis ; Phenotype ; Reactive Oxygen Species/metabolism ; Vaccines, DNA/immunology ; }, abstract = {Leishmaniases are severe vector-borne diseases affecting humans and animals, caused by Leishmania protozoans. Over one billion people and millions of dogs live in endemic areas for leishmaniases and are at risk of infection. Immune polarization plays a major role in determining the outcome of Leishmania infections: hosts displaying M1-polarized macrophages are protected, while those biased on the M2 side acquire a chronic infection that could develop into a deadly disease. The identification of the factors involved in M1 polarization is essential for the design of therapeutic and prophylactic interventions, including vaccines. Infection by the filarial nematode Dirofilaria immitis could be one of the factors that interfere with leishmaniasis in dogs. Indeed, filarial nematodes induce a partial skew of the immune response towards M1, likely caused by their bacterial endosymbionts, Wolbachia. Here we have examined the potential of Asaia[WSP], a bacterium engineered for the expression of the Wolbachia surface protein (WSP), as an inductor of M1 macrophage activation and Leishmania killing. Macrophages stimulated with Asaia[WSP] displayed a strong leishmanicidal activity, comparable to that determined by the choice-drug amphotericin B. Additionally, Asaia[WSP] determined the expression of markers of classical macrophage activation, including M1 cytokines, ROS and NO, and an increase in phagocytosis activity. Asaia not expressing WSP also induced macrophage activation, although at a lower extent compared to Asaia[WSP]. In summary, the results of the present study confirm the immunostimulating properties of WSP highlighting a potential therapeutic efficacy against Leishmania parasites. Furthermore, Asaia was designed as a delivery system for WSP, thus developing a novel type of immunomodulating agent, worthy of being investigated for immuno-prophylaxis and -therapy of leishmaniases and other diseases that could be subverted by M1 macrophage activation.}, } @article {pmid33158442, year = {2020}, author = {Pagendam, DE and Trewin, BJ and Snoad, N and Ritchie, SA and Hoffmann, AA and Staunton, KM and Paton, C and Beebe, N}, title = {Modelling the Wolbachia incompatible insect technique: strategies for effective mosquito population elimination.}, journal = {BMC biology}, volume = {18}, number = {1}, pages = {161}, pmid = {33158442}, issn = {1741-7007}, support = {1082127//National Health and Medical Research Council/International ; }, mesh = {Aedes/*microbiology ; Animals ; Female ; Male ; Markov Chains ; Models, Biological ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology ; Population Dynamics ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The Wolbachia incompatible insect technique (IIT) shows promise as a method for eliminating populations of invasive mosquitoes such as Aedes aegypti (Linnaeus) (Diptera: Culicidae) and reducing the incidence of vector-borne diseases such as dengue, chikungunya and Zika. Successful implementation of this biological control strategy relies on high-fidelity separation of male from female insects in mass production systems for inundative release into landscapes. Processes for sex-separating mosquitoes are typically error-prone and laborious, and IIT programmes run the risk of releasing Wolbachia-infected females and replacing wild mosquito populations.

RESULTS: We introduce a simple Markov population process model for studying mosquito populations subjected to a Wolbachia-IIT programme which exhibit an unstable equilibrium threshold. The model is used to study, in silico, scenarios that are likely to yield a successful elimination result. Our results suggest that elimination is best achieved by releasing males at rates that adapt to the ever-decreasing wild population, thus reducing the risk of releasing Wolbachia-infected females while reducing costs.

CONCLUSIONS: While very high-fidelity sex separation is required to avoid establishment, release programmes tend to be robust to the release of a small number of Wolbachia-infected females. These findings will inform and enhance the next generation of Wolbachia-IIT population control strategies that are already showing great promise in field trials.}, } @article {pmid33146464, year = {2021}, author = {Mugerwa, H and Wang, HL and Sseruwagi, P and Seal, S and Colvin, J}, title = {Whole-genome single nucleotide polymorphism and mating compatibility studies reveal the presence of distinct species in sub-Saharan Africa Bemisia tabaci whiteflies.}, journal = {Insect science}, volume = {28}, number = {6}, pages = {1553-1566}, pmid = {33146464}, issn = {1744-7917}, support = {Project/OPP1058938//African Cassava Whitefly/ ; OPP1080766//Enabling Research Tools for Cassava Virologists and Breeders/ ; /GATES/Bill & Melinda Gates Foundation/United States ; }, mesh = {Africa South of the Sahara ; Animals ; Genome, Insect ; Genome, Mitochondrial ; *Hemiptera/classification/genetics ; Manihot ; Phylogeny ; Plant Diseases ; *Polymorphism, Single Nucleotide ; }, abstract = {In sub-Saharan Africa cassava growing areas, two members of the Bemisia tabaci species complex termed sub-Saharan Africa 1 (SSA1) and SSA2 have been reported as the prevalent whiteflies associated with the spread of viruses that cause cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) pandemics. At the peak of CMD pandemic in the late 1990s, SSA2 was the prevalent whitefly, although its numbers have diminished over the last two decades with the resurgence of SSA1 whiteflies. Three SSA1 subgroups (SG1 to SG3) are the predominant whiteflies in East Africa and vary in distribution and biological properties. Mating compatibility between SSA1 subgroups and SSA2 whiteflies was reported as the possible driver for the resurgence of SSA1 whiteflies. In this study, a combination of both phylogenomic methods and reciprocal crossing experiments were applied to determine species status of SSA1 subgroups and SSA2 whitefly populations. Phylogenomic analyses conducted with 26 548 205 bp whole genome single nucleotide polymorphisms (SNPs) and the full mitogenomes clustered SSA1 subgroups together and separate from SSA2 species. Mating incompatibility between SSA1 subgroups and SSA2 further demonstrated their distinctiveness from each other. Phylogenomic analyses conducted with SNPs and mitogenomes also revealed different genetic relationships among SSA1 subgroups. The former clustered SSA1-SG1 and SSA1-SG2 together but separate from SSA1-SG3, while the latter clustered SSA1-SG2 and SSA1-SG3 together but separate from SSA1-SG1. Mating compatibility was observed between SSA1-SG1 and SSA1-SG2, while incompatibility occurred between SSA1-SG1 and SSA1-SG3, and SSA1-SG2 and SSA1-SG3. Mating results among SSA1 subgroups were coherent with phylogenomics results based on SNPs but not the full mitogenomes. Furthermore, this study revealed that the secondary endosymbiont-Wolbachia-did not mediate reproductive success in the crossing assays carried out. Overall, using genome wide SNPs together with reciprocal crossings assays, this study established accurate genetic relationships among cassava-colonizing populations, illustrating that SSA1 and SSA2 are distinct species while at least two species occur within SSA1 species.}, } @article {pmid33146105, year = {2020}, author = {Diarra, AZ and Kone, AK and Doumbo Niare, S and Laroche, M and Diatta, G and Atteynine, SA and Coulibaly, M and Sangare, AK and Kouriba, B and Djimde, A and Dabo, A and Sagara, I and Davoust, B and Ranque, S and Thera, MA and Raoult, D and Doumbo, OK and Parola, P}, title = {Molecular Detection of Microorganisms Associated with Small Mammals and Their Ectoparasites in Mali.}, journal = {The American journal of tropical medicine and hygiene}, volume = {103}, number = {6}, pages = {2542-2551}, pmid = {33146105}, issn = {1476-1645}, mesh = {Animals ; Bacteria/isolation & purification ; Disease Reservoirs ; Ectoparasitic Infestations/epidemiology/parasitology/*veterinary ; Mali/epidemiology ; Mites/microbiology ; Phylogeny ; Rodent Diseases/epidemiology/*parasitology ; Rodentia ; Siphonaptera/microbiology ; Ticks/microbiology ; Zoonoses ; }, abstract = {Small mammals are the natural reservoirs for many zoonotic pathogens. Using molecular tools, we assessed the prevalence of bacteria and protozoans in small mammals and their ectoparasites in Faladjè, Bougouni, and Bamoko, Mali. A total of 130 small mammals belonging to 10 different species were captured, of which 74 (56.9%) were infested by ectoparasites, including Laelaps echidnina, Xenopsylla cheopis, Amblyomma variegatum, Rhipicephalus sanguineus sensu lato, and Haemaphysalis spp. nymphs. DNA of Bartonella was found in 14/75 (18.7%), 6/48 (12.5%), and 3/7 (42.8%) small mammals from Faladjè, Bougouni, and Bamako, respectively. In Faladjè, Bartonella DNA was detected in 31/68 (45.6%) of L. echidnina and 14/22 (63.6%) of X. cheopis. In Bougouni, it was found in 2/26 (7.7%) of L. echidnina and 10/42 (23.8%) of X. cheopis. The sequences of Bartonella obtained from small mammals were close to those of Bartonella mastomydis, Bartonella elizabethae, and uncultured Bartonella spp. In Faladjè, Coxiella burnetii DNA was detected in 64.4% (29/45) of Haemaphysalis spp. ticks, 4.5% (2/44) of Mastomys erythroleucus, 12.5% (1/8) of Praomys daltoni, and 1.5% (1/68) of L. echidnina. We found DNA of Wolbachia in X. cheopis from Faladjè and DNA of Rickettsia africae and Ehrlichia ruminantium in Am. variegatum from Bougouni. The results of our study show that several small mammal species harbor and may serve as potential reservoirs of Bartonella spp., likely to play a major role in the maintenance, circulation, and potential transmission of bacteria in Mali. The pathogenicity of these bacteria for humans or animals remains to be demonstrated.}, } @article {pmid33144950, year = {2020}, author = {DeVries, ZC and Santangelo, RG and Booth, W and Lawrence, CG and Balvín, O and Bartonička, T and Schal, C}, title = {Reproductive compatibility among populations and host-associated lineages of the common bed bug (Cimex lectularius L.).}, journal = {Ecology and evolution}, volume = {10}, number = {20}, pages = {11090-11099}, pmid = {33144950}, issn = {2045-7758}, support = {P30 ES025128/ES/NIEHS NIH HHS/United States ; }, abstract = {As populations differentiate across geographic or host-association barriers, interpopulation fertility is often a measure of the extent of incipient speciation. The bed bug, Cimex lectularius L., was recently found to form two host-associated lineages within Europe: one found with humans (human-associated, HA) and the other found with bats (bat-associated, BA). No unequivocal evidence of contemporary gene flow between these lineages has been found; however, it is unclear whether this is due to an inability to produce viable "hybrid" offspring. To address this question and determine the extent of compatibility between host-associated lineages, we set up mating crosses among populations of bed bugs based on both their host association (human-HA vs. bat-BA) and geographic origin (North America vs. Europe). Within-population fecundity was significantly higher for all HA populations (>1.7 eggs/day) than for BA populations (<1 egg/day). However, all within-population crosses, regardless of host association, had >92% egg hatch rates. Contrary to previous reports, in all interlineage crosses, successful matings occurred, fertile eggs were oviposited, and the F1 "hybrid" generation was found to be reproductively viable. In addition, we evaluated interpopulation genetic variation in Wolbachia among host-associated lineages. We did not find any clear patterns related to host association, nor did we observe a homogenization of Wolbachia lineages across populations that might explain a breakdown of reproductive incompatibility. These results indicate that while the HA and BA populations of C. lectularius represent genetically differentiated host-associated lineages, possibly undergoing sympatric speciation, this is in its incipient stage as they remain reproductively compatible. Other behavioral, physiological, and/or ecological factors likely maintain host-associated differentiation.}, } @article {pmid33138055, year = {2020}, author = {Laidoudi, Y and Levasseur, A and Medkour, H and Maaloum, M and Ben Khedher, M and Sambou, M and Bassene, H and Davoust, B and Fenollar, F and Raoult, D and Mediannikov, O}, title = {An Earliest Endosymbiont, Wolbachia massiliensis sp. nov., Strain PL13 from the Bed Bug (Cimex hemipterus), Type Strain of a New Supergroup T.}, journal = {International journal of molecular sciences}, volume = {21}, number = {21}, pages = {}, pmid = {33138055}, issn = {1422-0067}, mesh = {Animals ; Bacterial Proteins/*genetics ; Bedbugs/*microbiology ; *Genome, Bacterial ; Genomics ; Phylogeny ; Symbiosis/*genetics ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {The symbiotic Wolbachia are the most sophisticated mutualistic bacterium among all insect-associated microbiota. Wolbachia-insect relationship fluctuates from the simple facultative/parasitic to an obligate nutritional-mutualistic association as it was the case of the bedbug-Wolbachia from Cimexlectularius. Understanding this association may help in the control of associated arthropods. Genomic data have proven to be reliable tools in resolving some aspects of these symbiotic associations. Although, Wolbachia appear to be fastidious or uncultivated bacteria which strongly limited their study. Here we proposed Drosophila S2 cell line for the isolation and culture model to study Wolbachia strains. We therefore isolated and characterized a novel Wolbachia strain associated with the bedbug Cimexhemipterus, designated as wChem strain PL13, and proposed Wolbachiamassiliensis sp. nov. strain wChem-PL13 a type strain of this new species from new supergroup T. Phylogenetically, T-supergroup was close to F and S-supergroups from insects and D-supergroup from filarial nematodes. We determined the 1,291,339-bp genome of wChem-PL13, which was the smallest insect-associated Wolbachia genomes. Overall, the wChem genome shared 50% of protein coding genes with the other insect-associated facultative Wolbachia strains. These findings highlight the diversity of Wolbachia genotypes as well as the Wolbachia-host relationship among Cimicinae subfamily. The wChem provides folate and riboflavin vitamins on which the host depends, while the bacteria had a limited translation mechanism suggesting its strong dependence to its hosts. However, the clear-cut distinction between mutualism and parasitism of the wChem in C. hemipterus cannot be yet ruled out.}, } @article {pmid33134332, year = {2020}, author = {Savić, S and Stosic, MZ and Marcic, D and Hernández, I and Potkonjak, A and Otasevic, S and Ruzic, M and Morchón, R}, title = {Seroepidemiological Study of Canine and Human Dirofilariasis in the Endemic Region of Northern Serbia.}, journal = {Frontiers in veterinary science}, volume = {7}, number = {}, pages = {571}, pmid = {33134332}, issn = {2297-1769}, abstract = {Dirofilariasis is a vector-borne zoonotic disease caused mainly by Dirofilaria immitis and Dirofilaria repens that affect dogs and humans all over the world. Serbia is considered an endemic country to both forms of dirofilariasis, although most of the population is concentrated in the north of the country. The aims of this study were to show the prevalence of D. immitis and D. repens in dogs and the seroprevalence in humans compared to previous studies in Northern Serbia. In total, 346 dog sera samples and 265 human samples were analyzed. Dog blood samples were analyzed using the modified Knott's method to check whether there were Dirofilaria spp. microfilariae and serum samples were checked by a commercial D. immitis antigen test. Human serum samples were analyzed with a non-commercial ELISA for detection of specific anti-D. immitis, anti-D. repens, and anti-Wolbachia IgG antibodies, and confirmed by western blotting. The overall prevalence for Dirofilaria spp. in dogs was 29.19%. The overall prevalence for D. immitis was 26.30%. The percentages of D. immitis and D. repens microfilaremia in dogs were 25.72 and 1.45%, respectively, while D. immitis./D. repens microfilaremia co-infections were also 1.45%. The overall seroprevalence for Dirofilaria spp. in humans was 3.77%. The overall seroprevalence for D. immitis was 1.51, 1.13% for D. repens, and for D. immitis/D. repens co-infections was 1.13%. The results indicate that D. immitis and D. repens are present in dogs and humans in the province of Vojvodina, in the northern part of Serbia. It is most likely associated with the presence of many rivers, the climate, and presence of mosquitoes in the area, so there could be a real public health risk.}, } @article {pmid33133140, year = {2020}, author = {Dacey, DP and Chain, FJJ}, title = {The Challenges of Microbial Control of Mosquito-Borne Diseases Due to the Gut Microbiome.}, journal = {Frontiers in genetics}, volume = {11}, number = {}, pages = {504354}, pmid = {33133140}, issn = {1664-8021}, abstract = {Mosquitoes are one of the deadliest animals on earth because of their ability to transmit a wide range of human pathogens. Traditional mosquito control methods use chemical insecticides, but with dwindling long-term effectiveness and negative effects on the environment, microbial forms of control have become common alternatives. The insecticide Bacillus thuringiensis subspecies israelensis (Bti) is the most popular of these alternatives, although it can also have direct effects on lowering environmental biodiversity and indirect effects on food-web relationships in the ecosystems where it is deployed. In addition, microbial control agents that impede pathogen development or transmission from mosquito to human are under investigation, including Wolbachia and Asaia, but unexpected interactions with mosquito gut bacteria can hinder their effectiveness. Improved characterization of mosquito gut bacterial communities is needed to determine the taxa that interfere with microbial controls and their effectiveness in wild populations. This mini-review briefly discusses relationships between mosquito gut bacteria and microbial forms of control, and the challenges in ensuring their success.}, } @article {pmid33128345, year = {2021}, author = {Sanaei, E and Charlat, S and Engelstädter, J}, title = {Wolbachia host shifts: routes, mechanisms, constraints and evolutionary consequences.}, journal = {Biological reviews of the Cambridge Philosophical Society}, volume = {96}, number = {2}, pages = {433-453}, doi = {10.1111/brv.12663}, pmid = {33128345}, issn = {1469-185X}, mesh = {Animals ; *Arthropods ; Biological Evolution ; Mosquito Vectors ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia is one of the most abundant endosymbionts on earth, with a wide distribution especially in arthropods. Effective maternal transmission and the induction of various phenotypes in their hosts are two key features of this bacterium. Here, we review our current understanding of another central aspect of Wolbachia's success: their ability to switch from one host species to another. We build on the proposal that Wolbachia host shifts occur in four main steps: (i) physical transfer to a new species; (ii) proliferation within that host; (iii) successful maternal transmission; and (iv) spread within the host species. Host shift can fail at each of these steps, and the likelihood of ultimate success is influenced by many factors. Some stem from traits of Wolbachia (different strains have different abilities for host switching), others on host features such as genetic resemblance (e.g. host shifting is likely to be easier between closely related species), ecological connections (the donor and recipient host need to interact), or the resident microbiota. Host shifts have enabled Wolbachia to reach its enormous current incidence and global distribution among arthropods in an epidemiological process shaped by loss and acquisition events across host species. The ability of Wolbachia to transfer between species also forms the basis of ongoing endeavours to control pests and disease vectors, following artificial introduction into uninfected hosts such as mosquitoes. Throughout, we emphasise the many knowledge gaps in our understanding of Wolbachia host shifts, and question the effectiveness of current methodology to detect these events. We conclude by discussing an apparent paradox: how can Wolbachia maintain its ability to undergo host shifts given that its biology seems dominated by vertical transmission?}, } @article {pmid33123758, year = {2021}, author = {Tzuri, N and Caspi-Fluger, A and Betelman, K and Rohkin Shalom, S and Chiel, E}, title = {Horizontal Transmission of Microbial Symbionts Within a Guild of Fly Parasitoids.}, journal = {Microbial ecology}, volume = {81}, number = {3}, pages = {818-827}, pmid = {33123758}, issn = {1432-184X}, support = {435/2018//Israel science Foundation/ ; }, mesh = {Animals ; Host-Parasite Interactions ; Larva ; Pupa ; *Wasps ; *Wolbachia/genetics ; }, abstract = {Many insects harbor facultative microbial symbionts which affect the ecology of their hosts in diverse ways. Most symbionts are transmitted vertically with high fidelity, whereas horizontal transmission occurs rarely. Parasitoid larvae feed on a single host and are in close physical contact with it, providing an ecological opportunity for symbionts' horizontal transmission, but there is little empirical evidence documenting this. Here we studied horizontal transmission of three bacterial symbionts-Rickettsia, Sodalis, and Wolbachia-between three fly pupal ectoparasitoid species: Spalangia cameroni, S. endius, and Muscidifurax raptor. Muscidifurax raptor readily parasitized and successfully developed on the Spalangia spp., while the inverse did not happen. The two Spalangia spp. attacked each other and conspecifics in very low rates. Symbiont horizontal transmissions followed by stable vertical transmission in the recipient species were achieved, in low percentages, only between conspecifics: Wolbachia from infected to uninfected M. raptor, Rickettsia in S. endius, and Sodalis in S. cameroni. Low frequency of horizontal transmissions occurred in the interspecific combinations, but none of them persisted in the recipient species beyond F4, at most. Our study is one of few to demonstrate symbionts' horizontal transmission between hosts within the same trophic level and guild and highlights the rarity of such events.}, } @article {pmid33122700, year = {2020}, author = {Chakraborty, A and Ashraf, MZ and Modlinger, R and Synek, J and Schlyter, F and Roy, A}, title = {Unravelling the gut bacteriome of Ips (Coleoptera: Curculionidae: Scolytinae): identifying core bacterial assemblage and their ecological relevance.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {18572}, pmid = {33122700}, issn = {2045-2322}, mesh = {Animals ; Bacteria/*classification/genetics/metabolism ; Coleoptera/metabolism/*microbiology ; Ecology ; Forests ; *Gastrointestinal Microbiome ; Nitrogen Fixation ; Phylogeny ; Trees/parasitology ; }, abstract = {Bark beetles often serve as forest damaging agents, causing landscape-level mortality. Understanding the biology and ecology of beetles are important for both, gathering knowledge about important forest insects and forest protection. Knowledge about the bark beetle gut-associated bacteria is one of the crucial yet surprisingly neglected areas of research with European tree-killing bark beetles. Hence, in this study, we survey the gut bacteriome from five Ips and one non-Ips bark beetles from Scolytinae. Results reveal 69 core bacterial genera among five Ips beetles that may perform conserved functions within the bark beetle holobiont. The most abundant bacterial genera from different bark beetle gut include Erwinia, Sodalis, Serratia, Tyzzerella, Raoultella, Rahnella, Wolbachia, Spiroplasma, Vibrio, and Pseudoxanthomonas. Notable differences in gut-associated bacterial community richness and diversity among the beetle species are observed. Furthermore, the impact of sampling location on the overall bark beetle gut bacterial community assemblage is also documented, which warrants further investigations. Nevertheless, our data expanded the current knowledge about core gut bacterial communities in Ips bark beetles and their putative function such as cellulose degradation, nitrogen fixation, detoxification of defensive plant compounds, and inhibition of pathogens, which could serve as a basis for further metatranscriptomics and metaproteomics investigations.}, } @article {pmid33120915, year = {2020}, author = {Allman, MJ and Fraser, JE and Ritchie, SA and Joubert, DA and Simmons, CP and Flores, HA}, title = {Wolbachia's Deleterious Impact on Aedes aegypti Egg Development: The Potential Role of Nutritional Parasitism.}, journal = {Insects}, volume = {11}, number = {11}, pages = {}, pmid = {33120915}, issn = {2075-4450}, support = {Research Training Program (RTP)//Department of Education, Skills and Employment, Australian Government/ ; }, abstract = {The artificial introduction of the endosymbiotic bacterium, Wolbachia pipientis, into Aedes (Ae.) aegypti mosquitoes reduces the ability of mosquitoes to transmit human pathogenic viruses and is now being developed as a biocontrol tool. Successful introgression of Wolbachia-carrying Ae. aegypti into native mosquito populations at field sites in Australia, Indonesia and Malaysia has been associated with reduced disease prevalence in the treated community. In separate field programs, Wolbachia is also being used as a mosquito population suppression tool, where the release of male only Wolbachia-infected Ae. aegypti prevents the native mosquito population from producing viable eggs, subsequently suppressing the wild population. While these technologies show great promise, they require mass rearing of mosquitoes for implementation on a scale that has not previously been done. In addition, Wolbachia induces some negative fitness effects on Ae. aegypti. While these fitness effects differ depending on the Wolbachia strain present, one of the most consistent and significant impacts is the shortened longevity and viability of eggs. This review examines the body of evidence behind Wolbachia's negative effect on eggs, assesses nutritional parasitism as a key cause and considers how these impacts could be overcome to achieve efficient large-scale rearing of these mosquitoes.}, } @article {pmid33108676, year = {2021}, author = {Prigot-Maurice, C and de Cerqueira De Araujo, A and Beltran-Bech, S and Braquart-Varnier, C}, title = {Immune priming depends on age, sex and Wolbachia in the interaction between Armadillidium vulgare and Salmonella.}, journal = {Journal of evolutionary biology}, volume = {34}, number = {2}, pages = {256-269}, doi = {10.1111/jeb.13721}, pmid = {33108676}, issn = {1420-9101}, mesh = {Age Factors ; Animals ; Female ; Host-Pathogen Interactions/*immunology ; Isopoda/*immunology ; Male ; Salmonella enterica/*physiology ; Sex Factors ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The protection conferred by a first infection upon a second pathogenic exposure (i.e. immune priming) is an emergent research topic in the field of invertebrate immunity. Immune priming has been demonstrated in various species, but little is known about the intrinsic factors that may influence this immune process. In this study, we tested whether age, gender and the symbiotic bacterium Wolbachia affect the protection resulting from immune priming in A. vulgare against S. enterica. We firstly primed young and old, symbiotic and asymbiotic males and females, either with a non-lethal low dose of S. enterica, LB broth or without injection (control). Seven days post-injection, we performed a LD50 injection of S. enterica in all individuals and we monitored their survival rates. We demonstrated that survival capacities depend on these three factors: young and old asymbiotic individuals (males and females) expressed immune priming (S. enterica-primed individuals survived better than LB-primed and non-primed), with a general decline in the strength of protection in old females, but not in old males, compared to young. When Wolbachia is present, the immune priming protection was observed in old, but not in young symbiotic individuals, even if the Wolbachia load on entire individuals is equivalent regardless to age. Our overall results showed that the immune priming protection in A. vulgare depends on individuals' states, highlighting the need to consider these factors both in mechanistical and evolutionary studies focusing on invertebrate's immunity.}, } @article {pmid33099649, year = {2021}, author = {Sandri, TL and Kreidenweiss, A and Cavallo, S and Weber, D and Juhas, S and Rodi, M and Woldearegai, TG and Gmeiner, M and Veletzky, L and Ramharter, M and Tazemda-Kuitsouc, GB and Matsiegui, PB and Mordmüller, B and Held, J}, title = {Molecular Epidemiology of Mansonella Species in Gabon.}, journal = {The Journal of infectious diseases}, volume = {223}, number = {2}, pages = {287-296}, doi = {10.1093/infdis/jiaa670}, pmid = {33099649}, issn = {1537-6613}, mesh = {Animals ; Carrier State/parasitology ; Cross-Sectional Studies ; Gabon/epidemiology ; Humans ; Loa/genetics ; Male ; Mansonella/*classification/*genetics ; Mansonelliasis/*epidemiology/*parasitology ; Molecular Epidemiology ; Polymerase Chain Reaction ; Rural Population ; }, abstract = {Mansonella perstans, a filarial nematode, infects large populations in Africa and Latin America. Recently, a potential new species, Mansonella sp "DEUX," was reported. Carriage of endosymbiotic Wolbachia opens treatment options for Mansonella infections. Within a cross-sectional study, we assessed the prevalence of filarial infections in 834 Gabonese individuals and the presence of the endosymbiont Wolbachia. Almost half of the participants (400/834 [48%]) were infected with filarial nematodes, with Mansonella sp "DEUX" being the most frequent (295/400 [74%]), followed by Loa loa (273/400 [68%]) and Mansonella perstans (82/400 [21%]). Being adult/elderly, male, and living in rural areas was associated with a higher risk of infection. Wolbachia carriage was confirmed in M. perstans and Mansonella sp "DEUX." In silico analysis revealed that Mansonella sp "DEUX" is not detected with currently published M. perstans-specific assays. Mansonella infections are highly prevalent in Gabon and might have been underreported, likely also beyond Gabon.}, } @article {pmid33095791, year = {2020}, author = {Knerer, G and Currie, CSM and Brailsford, SC}, title = {The economic impact and cost-effectiveness of combined vector-control and dengue vaccination strategies in Thailand: results from a dynamic transmission model.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {10}, pages = {e0008805}, pmid = {33095791}, issn = {1935-2735}, mesh = {Aedes/microbiology/physiology/virology ; Animals ; Cost-Benefit Analysis ; Dengue/*economics/epidemiology/prevention & control/transmission ; Humans ; Mosquito Control/*economics/*legislation & jurisprudence ; Mosquito Vectors/microbiology/physiology/virology ; Thailand/epidemiology ; Vaccination/*economics ; Wolbachia/physiology ; }, abstract = {BACKGROUND AND AIMS: Dengue fever is a major public health problem in tropical/subtropical regions. Prior economic analyses have predominantly evaluated either vaccination or vector-control programmes in isolation and do not really consider the incremental benefits and cost-effectiveness of mixed strategies and combination control. We estimated the cost-effectiveness of single and combined approaches in Thailand.

METHODS: The impacts of different control interventions were analysed using a previously published mathematical model of dengue epidemiology and control incorporating seasonality, age structure, consecutive infection, cross protection, immune enhancement and combined vector-host transmission. An economic model was applied to simulation results to estimate the cost-effectiveness of 4 interventions and their various combinations (6 strategies): i) routine vaccination of 1-year olds; ii) chemical vector control strategies targeting adult and larval stages separately; iii) environmental management/ public health education and awareness [EM/ PHEA]). Payer and societal perspectives were considered. The health burden of dengue fever was assessed using disability-adjusted life-years (DALYs) lost. Costs and effects were assessed for 10 years. Costs were discounted at 3% annually and updated to 2013 United States Dollars. Incremental cost-effectiveness analysis was carried out after strategies were rank-ordered by cost, with results presented in a table of incremental analysis. Sensitivity and scenario analyses were undertaken; and the impact and cost-effectiveness of Wolbachia was evaluated in exploratory scenario analyses.

RESULTS: From the payer and societal perspectives, 2 combination strategies were considered optimal, as all other control strategies were dominated. Vaccination plus adulticide plus EM/ PHEA was deemed cost-effective according to multiple cost-effectiveness criteria. From the societal perspective, incremental differences vs. adulticide and EM/ PHEA resulted in costs of $157.6 million and DALYs lost of 12,599, giving an expected ICER of $12,508 per DALY averted. Exploratory scenario analyses showed Wolbachia to be highly cost-effective ($343 per DALY averted) vs. other single control measures.

CONCLUSIONS: Our model shows that individual interventions can be cost-effective, but that important epidemiological reductions and economic impacts are demonstrated when interventions are combined as part of an integrated approach to combating dengue fever. Exploratory scenario analyses demonstrated the potential epidemiological and cost-effective impact of Wolbachia when deployed at scale on a nationwide basis. Our findings were robust in the face of sensitivity analyses.}, } @article {pmid33083739, year = {2020}, author = {Huang, B and Yang, Q and Hoffmann, AA and Ritchie, SA and van den Hurk, AF and Warrilow, D}, title = {Wolbachia Genome Stability and mtDNA Variants in Aedes aegypti Field Populations Eight Years after Release.}, journal = {iScience}, volume = {23}, number = {10}, pages = {101572}, pmid = {33083739}, issn = {2589-0042}, abstract = {A dengue suppression strategy based on release of Aedes aegypti mosquitoes infected with the bacterium Wolbachia pipientis is being trialed in many countries. Wolbachia inhibits replication and transmission of dengue viruses. Questions remain regarding the long-term stability of virus-suppressive effects. We sequenced the Wolbachia genome and analyzed Ae. aegypti mitochondrial DNA markers isolated from mosquitoes sampled 2-8 years after releases in the greater Cairns region, Australia. Few changes were detected when Wolbachia genomes of field mosquitoes were compared with Wolbachia genomes of mosquitoes obtained soon after initial releases. Mitochondrial variants associated with the initial Wolbachia release stock are now the only variants found in release sites, highlighting maternal leakage as a possible explanation for rare Wolbachia-negative mosquitoes and not migration from non-release areas. There is no evidence of changes in the Wolbachia genome that indicate selection against its viral-suppressive effects or other phenotypes attributable to infection with the bacterium.}, } @article {pmid33081703, year = {2020}, author = {Duplouy, A and Pranter, R and Warren-Gash, H and Tropek, R and Wahlberg, N}, title = {Towards unravelling Wolbachia global exchange: a contribution from the Bicyclus and Mylothris butterflies in the Afrotropics.}, journal = {BMC microbiology}, volume = {20}, number = {1}, pages = {319}, pmid = {33081703}, issn = {1471-2180}, support = {790531//H2020 Marie Skłodowska-Curie Actions/ ; 1328944//Academy of Finland/ ; UNCE204069//Charles University/ ; PRIMUS/17/SCI/8//Charles University/ ; Waitt Grant W163-11//National Geographic Society/ ; 2015-04441//Swedish Research Council/ ; }, mesh = {Africa ; Animals ; Biodiversity ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Geography ; Lepidoptera/genetics/*physiology ; Phylogeny ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Phylogenetically closely related strains of maternally inherited endosymbiotic bacteria are often found in phylogenetically divergent, and geographically distant insect host species. The interspecies transfer of the symbiont Wolbachia has been thought to have occurred repeatedly, facilitating its observed global pandemic. Few ecological interactions have been proposed as potential routes for the horizontal transfer of Wolbachia within natural insect communities. These routes are however likely to act only at the local scale, but how they may support the global distribution of some Wolbachia strains remains unclear.

RESULTS: Here, we characterize the Wolbachia diversity in butterflies from the tropical forest regions of central Africa to discuss transfer at both local and global scales. We show that numerous species from both the Mylothris (family Pieridae) and Bicyclus (family Nymphalidae) butterfly genera are infected with similar Wolbachia strains, despite only minor interclade contacts across the life cycles of the species within their partially overlapping ecological niches. The phylogenetic distance and differences in resource use between these genera rule out the role of ancestry, hybridization, and shared host-plants in the interspecies transfer of the symbiont. Furthermore, we could not identify any shared ecological factors to explain the presence of the strains in other arthropod species from other habitats, or even ecoregions.

CONCLUSION: Only the systematic surveys of the Wolbachia strains from entire species communities may offer the material currently lacking for understanding how Wolbachia may transfer between highly different and unrelated hosts, as well as across environmental scales.}, } @article {pmid33075316, year = {2020}, author = {Wangkeeree, J and Suwanchaisri, K and Roddee, J and Hanboonsong, Y}, title = {Effect of Wolbachia infection states on the life history and reproductive traits of the leafhopper Yamatotettix flavovittatus Matsumura.}, journal = {Journal of invertebrate pathology}, volume = {177}, number = {}, pages = {107490}, doi = {10.1016/j.jip.2020.107490}, pmid = {33075316}, issn = {1096-0805}, mesh = {Animals ; Female ; Hemiptera/growth & development/microbiology/*physiology ; *Life History Traits ; Male ; Nymph/growth & development/microbiology/physiology ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a maternally inherited bacterium of insects that can affect host reproduction and fitness. We examined the effect of Wolbachia infection on the life history and reproductive traits of the leafhopper Yamatotettix flavovittatus, which is a vector of the phytoplasma that causes white leaf disease in sugarcane. This investigation was performed using Wolbachia-infected and uninfected leafhopper lineages. Results revealed that Wolbachia infection did not significantly affect the survival of nymphal stages, male longevity, and sex ratio. However, Wolbachia-infected lineages had prolonged immature development periods and female longevity. In intrapopulation crosses, Wolbachia infection had no significant effects on occupation success, number of eggs laid, and female offspring, but the effect on egg-hatching varied. In interpopulation crosses, Wolbachia infection had no significant effect on occupation success and female offspring, but it did affect the number of eggs laid and egg-hatching rates. Assortative pairings regarding infection status resulted in normal egg deposition and hatching, whereas disassortative pairings resulted in lower egg deposition and no hatching. Wolbachia was thus shown to be highly vertically transmitted (>98% of the tested individuals). Our findings provide additional data on the interactions between Wolbachia in insect hosts. This evidence of perfect maternal transmission and strong reproductive incompatibility highlights the importance of further studies on the use of Wolbachia as a biological control agent for the leafhopper vector.}, } @article {pmid33073851, year = {2020}, author = {Can-Vargas, X and Barboza, N and Fuchs, EJ and Hernández, EJ}, title = {Spatial Distribution of Whitefly Species (Hemiptera: Aleyrodidae) and Identification of Secondary Bacterial Endosymbionts in Tomato Fields in Costa Rica.}, journal = {Journal of economic entomology}, volume = {113}, number = {6}, pages = {2900-2910}, pmid = {33073851}, issn = {1938-291X}, mesh = {Animals ; Bacteria ; Costa Rica ; *Hemiptera/genetics ; *Solanum lycopersicum ; Symbiosis ; }, abstract = {In Costa Rica, tomato (Solanum lycopersicum Linnaeus) Linnaeus (Solanales: Solanaceae) is one of the crops most severely affected by the whiteflies (Hemiptera: Aleyrodidae) Trialeurodes vaporariorum (Westwood) and the Bemisia tabaci (Gennadius) species complex. The objective of this study was to monitor the spatial distribution and diversity of these species and to detect the presence of secondary bacterial endosymbionts in individuals collected in areas of intensive tomato production. In total, 628 whitefly individuals were identified to the species level using restriction analysis (PCR-RFLP) of a fragment of the mitochondrial cytochrome C oxidase I gene (mtCOI). Trialeurodes vaporariorum was the predominant species, followed by B. tabaci Mediterranean (MED). Bemisia tabaci New World (NW) and B. tabaci Middle East-Asia Minor 1 (MEAM1) were present in lower numbers. The mtCOI fragment was sequenced for 89 individuals and a single haplotype was found for each whitefly species. Using molecular markers, the 628 individuals were analyzed for the presence of four endosymbionts. Arsenophonus Gherna et al. (Enterobacterales: Morganellaceae) was most frequently associated with T. vaporariorum, whereas Wolbachia Hertig (Rickettsiales: Anaplasmataceae) and Rickettsia da Rocha-Lima (Rickettsiales: Rickettsiaceae) were associated with B. tabaci MED. This study confirmed that B. tabaci NW has not been completely displaced by the invasive species B. tabaci MED and B. tabaci MEAM1 present in the country. An association was found between whitefly species present in tomato and certain secondary endosymbionts, elevation was the most likely environmental factor to affect their frequency.}, } @article {pmid33070738, year = {2020}, author = {Verspoor, RL and Price, TAR and Wedell, N}, title = {Selfish genetic elements and male fertility.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {375}, number = {1813}, pages = {20200067}, pmid = {33070738}, issn = {1471-2970}, mesh = {Animals ; Fertility/*genetics ; Humans ; Male ; Repetitive Sequences, Nucleic Acid/*physiology ; Spermatozoa/*physiology ; }, abstract = {Selfish genetic elements (SGEs) are diverse and near ubiquitous in Eukaryotes and can be potent drivers of evolution. Here, we discuss SGEs that specifically act on sperm to gain a transmission advantage to the next generation. The diverse SGEs that affect sperm often impose costs on carrier males, including damaging ejaculates, skewing offspring sex ratios and in particular reducing sperm-competitive success of SGE-carrying males. How males and females tolerate and mitigate against these costs is a dynamic and expanding area of research. The intense intra-genomic conflict that these selfish elements generate could also have implications for male fertility and spermatogenesis more widely. This article is part of the theme issue 'Fifty years of sperm competition'.}, } @article {pmid33070212, year = {2021}, author = {Xu, S and Jiang, L and Qiao, G and Chen, J}, title = {Diversity of bacterial symbionts associated with Myzus persicae (Sulzer) (Hemiptera: Aphididae: Aphidinae) revealed by 16S rRNA Illumina sequencing.}, journal = {Microbial ecology}, volume = {81}, number = {3}, pages = {784-794}, pmid = {33070212}, issn = {1432-184X}, support = {2016YFE0203100//National Key R & D Program of China/ ; 31620103916//National Natural Science Foundation of China (CN)/ ; 31772492//National Natural Science Foundation of China/ ; XDA19050303//Strategic Priority Research Program of the Chinese Academy of Sciences/ ; 2020087//Youth Innovation Promotion Association of Chinese Academy of Sciences/ ; }, mesh = {Animals ; *Aphids ; Bacteria/genetics ; *Buchnera/genetics ; High-Throughput Nucleotide Sequencing ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; }, abstract = {Aphids are known to be associated with a variety of symbiotic bacteria. To improve our knowledge of the bacterial diversity of polyphagous aphids, in the present study, we investigated the microbiota of the cosmopolitan agricultural pest Myzus persicae (Sulzer). Ninety-two aphid samples collected from different host plants in various regions of China were examined using high-throughput amplicon sequencing. We comprehensively characterized the symbiont diversity of M. persicae and assessed the variations in aphid-associated symbiont communities. We detected a higher diversity of symbionts than has been previously observed. M. persicae hosted the primary endosymbiont Buchnera aphidicola and seven secondary symbionts, among which Wolbachia was the most prevalent and Rickettsia, Arsenophonus, and Spiroplasma were reported for the first time. Ordination analyses and statistical tests revealed that the symbiont flora associated with M. persicae did not change with respect to host plant or geography, which may be due to frequent migrations between different aphid populations. These findings will advance our knowledge of the microbiota of polyphagous insects and will enrich our understanding of assembly of host-microbiome systems.}, } @article {pmid33067437, year = {2020}, author = {Scholz, M and Albanese, D and Tuohy, K and Donati, C and Segata, N and Rota-Stabelli, O}, title = {Large scale genome reconstructions illuminate Wolbachia evolution.}, journal = {Nature communications}, volume = {11}, number = {1}, pages = {5235}, pmid = {33067437}, issn = {2041-1723}, mesh = {Evolution, Molecular ; Gene Transfer, Horizontal ; *Genome, Bacterial ; Host Specificity ; Phylogeny ; Wolbachia/classification/*genetics/physiology ; }, abstract = {Wolbachia is an iconic example of a successful intracellular bacterium. Despite its importance as a manipulator of invertebrate biology, its evolutionary dynamics have been poorly studied from a genomic viewpoint. To expand the number of Wolbachia genomes, we screen over 30,000 publicly available shotgun DNA sequencing samples from 500 hosts. By assembling over 1000 Wolbachia genomes, we provide a substantial increase in host representation. Our phylogenies based on both core-genome and gene content provide a robust reference for future studies, support new strains in model organisms, and reveal recent horizontal transfers amongst distantly related hosts. We find various instances of gene function gains and losses in different super-groups and in cytoplasmic incompatibility inducing strains. Our Wolbachia-host co-phylogenies indicate that horizontal transmission is widespread at the host intraspecific level and that there is no support for a general Wolbachia-mitochondrial synchronous divergence.}, } @article {pmid33066069, year = {2020}, author = {Pan, X and Wang, X and Zhang, F}, title = {New Insights into Cockroach Control: Using Functional Diversity of Blattella germanica Symbionts.}, journal = {Insects}, volume = {11}, number = {10}, pages = {}, pmid = {33066069}, issn = {2075-4450}, support = {81572027//National Natural Science Foundation of China/ ; 2017GSF221012//the Key Programs for Science and Technology Development of Shandong Province/ ; 2017ZX100303404//the National Critical Project for Science and Technology on Infectious Diseases of P. R. China/ ; }, abstract = {Insects have close symbiotic relationships with several microbes, which extends the limited metabolic networks of most insects. Using symbiotic microorganisms for the biological control of pests and insect-borne diseases has become a promising direction. Blattella germanica (L.) (Blattaria: Blattidae) is a public health pest worldwide, which is difficult to control because of its strong reproductive ability, adaptability, and resistance to insecticides. In this paper, the diverse biological functions (nutrition, reproductive regulation, insecticide resistance, defense, and behavior) of symbionts were reviewed, and new biological control strategies on the basis of insect-symbiont interaction were proposed. We highlight new directions in B. germanica control, such as suppressing cockroach population using Wolbachia or paratransgenes, and combining fungal insecticides with synergistic agents to enhance insecticidal efficacy.}, } @article {pmid33053771, year = {2020}, author = {Lee, CC and Lin, CY and Tseng, SP and Matsuura, K and Yang, CS}, title = {Ongoing Coevolution of Wolbachia and a Widespread Invasive Ant, Anoplolepis gracilipes.}, journal = {Microorganisms}, volume = {8}, number = {10}, pages = {}, pmid = {33053771}, issn = {2076-2607}, abstract = {While Wolbachia are commonly found among arthropods, intraspecific infection rates can vary substantially across the geographic populations. Here we report nearly 100% prevalence of Wolbachia in the global populations of the yellow crazy ant, Anoplolepis gracilipes. To understand coevolutionary history between Wolbachia and A. gracilipes, we identified single nucleotide polymorphisms (SNPs) in Wolbachia from the ant across 12 geographical regions and compared the phylogeny of SNP-based Wolbachia to patterns of the ant's mitochondrial DNA (mtDNA) variation. Our results revealed a strong concordance between phylogenies of Wolbachia and host mtDNA, providing immediate evidence of co-divergence. Among eight identified SNP loci separating the genetic clusters of Wolbachia, seven loci are located in potential protein-coding genes, three of which being non-synonymous SNPs that may influence gene functions. We found a Wolbachia hypothetical protein gene with signature of positive selection. These findings jointly allow us to characterize Wolbachia-ant coevolution and also raise a question about mechanism(s) underlying maintenance of high prevalence of Wolbachia during the colonization of this invasive ant.}, } @article {pmid33050570, year = {2020}, author = {Montoya-Alonso, JA and Morchón, R and Matos, JI and Falcón-Cordón, Y and Costa-Rodriguez, N and Carretón, E}, title = {Dirofilaria immitis Could Be a Risk Factor for the Development of Allergic Diseases in Humans.}, journal = {Animals : an open access journal from MDPI}, volume = {10}, number = {10}, pages = {}, pmid = {33050570}, issn = {2076-2615}, support = {ProID2017010111//Agencia Canaria de Investigación, Innovación y Sociedad de la Información/ ; }, abstract = {The Canary Islands are hyperendemic for canine heartworm and the risk of zoonotic infection is high. Additionally, there is evidence of development of allergies due to nematode infections in animals and humans. Since the population of the Canary Islands presents high prevalence of allergic comorbidities, and previous studies have shown a possible relationship between allergies and seropositivity to heartworm, the aim was to evaluate whether exposure to heartworm may contribute to the development of allergies in the human population. First, an epidemiological study was conducted on 248 owners of dogs with/without heartworm infection in the Canary Islands. Secondly, a serological analysis of the presence of total IgE and specific IgE against heartworm was conducted in 66 samples of inhabitants of the Canary Islands. The survey showed that allergic owners had heartworm-positive dogs more frequently (p < 0.01). Of the analyzed human samples, 43.9% were seropositive to heartworm and Wolbachia. Total IgE concentrations were increased in 34.5% seropositive samples and 8.1% seronegative samples. Specific IgE against heartworm was only found in human seropositive samples (17.2%). Contact with heartworm may be a risk factor for the development of allergic diseases, although further studies to elucidate the relationship between heartworm and allergies should be carried out.}, } @article {pmid33048997, year = {2020}, author = {Carneiro Dutra, HL and Deehan, MA and Frydman, H}, title = {Wolbachia and Sirtuin-4 interaction is associated with alterations in host glucose metabolism and bacterial titer.}, journal = {PLoS pathogens}, volume = {16}, number = {10}, pages = {e1008996}, pmid = {33048997}, issn = {1553-7374}, mesh = {Adenosine Triphosphate/genetics/metabolism ; Animals ; Carbohydrate Metabolism ; Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster ; Gene Expression Regulation ; Glucose/*metabolism ; Glutamate Dehydrogenase/genetics/metabolism ; Host-Pathogen Interactions ; Longevity ; Reproduction ; Signal Transduction ; Sirtuins/*metabolism ; Symbiosis ; Wolbachia/metabolism/*physiology ; }, abstract = {Wolbachia is an intracellular bacterial symbiont of arthropods notorious for inducing many reproductive manipulations that foster its dissemination. Wolbachia affects many aspects of host biology, including metabolism, longevity and physiology, being described as a nutrient provisioning or metabolic parasite, depending on the host-microbe association. Sirtuins (SIRTs) are a family of NAD+-dependent post-translational regulatory enzymes known to affect many of the same processes altered by Wolbachia, including aging and metabolism, among others. Despite a clear overlap in control of host-derived pathways and physiology, no work has demonstrated a link between these two regulators. We used genetically tractable Drosophila melanogaster to explore the role of sirtuins in shaping signaling pathways in the context of a host-symbiont model. By using transcriptional profiling and metabolic assays in the context of genetic knockouts/over-expressions, we examined the effect of several Wolbachia strains on host sirtuin expression across distinct tissues and timepoints. We also quantified the downstream effects of the sirtuin x Wolbachia interaction on host glucose metabolism, and in turn, how it impacted Wolbachia titer. Our results indicate that the presence of Wolbachia is associated with (1) reduced sirt-4 expression in a strain-specific manner, and (2) alterations in host glutamate dehydrogenase expression and ATP levels, key components of glucose metabolism. We detected high glucose levels in Wolbachia-infected flies, which further increased when sirt-4 was over-expressed. However, under sirt-4 knockout, flies displayed a hypoglycemic state not rescued to normal levels in the presence of Wolbachia. Finally, whole body sirt-4 over-expression resulted in reduced Wolbachia ovarian titer. Our results expand knowledge of Wolbachia-host associations in the context of a yet unexplored class of host post-translational regulatory enzymes with implications for conserved host signaling pathways and bacterial titer, factors known to impact host biology and the symbiont's ability to spread through populations.}, } @article {pmid33035486, year = {2020}, author = {Gong, JT and Li, Y and Li, TP and Liang, Y and Hu, L and Zhang, D and Zhou, CY and Yang, C and Zhang, X and Zha, SS and Duan, XZ and Baton, LA and Hong, XY and Hoffmann, AA and Xi, Z}, title = {Stable Introduction of Plant-Virus-Inhibiting Wolbachia into Planthoppers for Rice Protection.}, journal = {Current biology : CB}, volume = {30}, number = {24}, pages = {4837-4845.e5}, doi = {10.1016/j.cub.2020.09.033}, pmid = {33035486}, issn = {1879-0445}, mesh = {Animals ; Crop Protection/*methods ; Feasibility Studies ; Hemiptera/*microbiology/virology ; Insect Vectors/*microbiology ; Oryza/parasitology/*virology ; Plant Diseases/prevention & control/virology ; Reoviridae/pathogenicity ; Wolbachia/*pathogenicity ; }, abstract = {Progress has been made in developing the maternally inherited endosymbiotic bacterium Wolbachia as a tool for protecting humans from mosquito-borne diseases. In contrast, Wolbachia-based approaches have not yet been developed for the protection of plants from insect pests and their associated diseases, with a major challenge being the establishment of artificial Wolbachia infections expressing desired characteristics in the hemipterans that transmit the majority of plant viruses. Here, we report stable introduction of Wolbachia into the brown planthopper, Nilaparvata lugens, the most destructive rice pest that annually destroys millions of hectares of staple crops. The Wolbachia strain wStri from the small brown planthopper, Laodelphax striatellus, was transferred to this new host, where it showed high levels of cytoplasmic incompatibility, enabling rapid invasion of laboratory populations. Furthermore, wStri inhibited infection and transmission of Rice ragged stunt virus and mitigated virus-induced symptoms in rice plants, opening up the development of Wolbachia-based strategies against major agricultural pests and their transmitted pathogens. VIDEO ABSTRACT.}, } @article {pmid33034342, year = {2020}, author = {Puerta-Guardo, H and Contreras-Perera, Y and Perez-Carrillo, S and Che-Mendoza, A and Ayora-Talavera, G and Vazquez-Prokopec, G and Martin-Park, A and Zhang, D and Manrique-Saide, P and , }, title = {Wolbachia in Native Populations of Aedes albopictus (Diptera: Culicidae) From Yucatan Peninsula, Mexico.}, journal = {Journal of insect science (Online)}, volume = {20}, number = {5}, pages = {}, pmid = {33034342}, issn = {1536-2442}, mesh = {Aedes/*microbiology ; Animals ; Mexico ; Mosquito Vectors/microbiology ; Pathology, Molecular ; Wolbachia/*isolation & purification ; }, abstract = {This study reports the results of a molecular screening for Wolbachia (Wb) infection in Aedes albopictus (Skuse) populations recently established in the Yucatan Peninsula, Mexico. To do so, collections of free-flying adults with BG traps and emerged adults from eggs after ovitrap field collections were performed in three suburban localities of the city of Merida, Yucatan. Overall, local populations of Ae. albopictus present a natural Wb infection rate of ~40% (18 of 45). Wb infection was detected in both field-collected adults (76.5%, 13 of 17) and eggs reared (17.8%, 5 of 28) and in 37.9% (11/29) of females and 43.7% (7/16) of male Ae. albopictus mosquitoes. An initial screening for Wolbachia strain typing showed that native Ae. albopictus were naturally coinfected with both wAlbA and wAlbB strains. The knowledge of the prevalence and diversity of Wolbachia strains in local populations of Aedes mosquitoes is part of the baseline information required for current and future Wolbachia-based vector control approaches to be conducted in Mexico.}, } @article {pmid33033285, year = {2020}, author = {Ogunlade, ST and Adekunle, AI and Meehan, MT and Rojas, DP and McBryde, ES}, title = {Modeling the potential of wAu-Wolbachia strain invasion in mosquitoes to control Aedes-borne arboviral infections.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {16812}, pmid = {33033285}, issn = {2045-2322}, mesh = {Aedes/microbiology/physiology/*virology ; Animals ; Arbovirus Infections/*prevention & control ; Female ; Male ; Models, Biological ; Pest Control, Biological/*methods ; Reproduction ; *Wolbachia ; }, abstract = {Arboviral infections such as dengue, Zika and chikungunya are fast spreading diseases that pose significant health problems globally. In order to control these infections, an intracellular bacterium called Wolbachia has been introduced into wild-type mosquito populations in the hopes of replacing the vector transmitting agent, Aedes aegypti with one that is incapable of transmission. In this study, we developed a Wolbachia transmission model for the novel wAu strain which possesses several favourable traits (e.g., enhanced viral blockage and maintenance at higher temperature) but not cyctoplasmic incompatibility (CI)-when a Wolbachia-infected male mosquito mates with an uninfected female mosquito, producing no viable offspring. This model describes the competitive dynamics between wAu-Wolbachia-infected and uninfected mosquitoes and the role of imperfect maternal transmission. By analysing the system via computing the basic reproduction number(s) and stability properties, the potential of the wAu strain as a viable strategy to control arboviral infections is established. The results of this work show that enhanced maintenance of Wolbachia infection at higher temperatures can overcome the lack of CI induction to support wAu-Wolbachia infected mosquito invasion. This study will support future arboviral control programs, that rely on the introduction of new Wolbachia variants.}, } @article {pmid33032591, year = {2020}, author = {Nguyen, AHL and Tiawsirisup, S and Kaewthamasorn, M}, title = {Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand.}, journal = {BMC veterinary research}, volume = {16}, number = {1}, pages = {380}, pmid = {33032591}, issn = {1746-6148}, support = {STF6101331002-1//Chulalongkorn University/ ; Second Century Fund//Chulalongkorn University/ ; GRU 6203331007-1//Chulalongkorn University/ ; Mid-Career Research Grant 2020//National Research Council of Thailand/ ; }, mesh = {Anaplasma/genetics/isolation & purification ; Anaplasma marginale/genetics/isolation & purification ; Anaplasmataceae/classification/genetics/*isolation & purification ; Anaplasmosis/*epidemiology/microbiology ; Animals ; Buffaloes/*microbiology ; Female ; Male ; RNA, Ribosomal, 16S/genetics ; Sex Factors ; Thailand/epidemiology ; }, abstract = {BACKGROUND: Anaplasmosis, an animal disease caused by rickettsial bacteria in the genus Anaplasma, is of considerable economic importance in livestock animals in many countries worldwide. The objectives of this study were to determine the identity, prevalence, and geographic distribution of Ehrlichia and Anaplasma in naturally infected water buffalo in Thailand using PCR amplification and sequencing of the 16S ribosomal RNA and heat shock protein groEL genes. A total of 456 buffalo blood samples from Thailand were investigated. Species identification and genetic differentiation of intra-population and inter-population with the global isolates were conducted based on nucleotide sequences. Interplay between the infection and host factors was also assessed.

RESULTS: Overall, 41% of water buffalo were found to be infected with rickettsial organisms in the family Anaplasmataceae, but Ehrlichia spp., Neorickettsia spp., and Wolbachia spp. were not found in any of the sequenced samples in this study. Female buffalo were more frequently infected with bacteria in the family Anaplasmataceae than males [71 out of 176 females (40.3%) versus 11 out of 47 males (23.4%)]. The Odds Ratio value indicated that the risk of infection for female buffalo was 2.2-fold higher than that for males (p < 0.05). We detected three haplotypes of A. marginale 16S rRNA gene and they were placed in a clade that was closely related to the A. marginale in buffalo in China; and cattle in Thailand, Uganda, and China. Homology searching of groEL sequences against the GenBank™ database using the BLASTn algorithm revealed that the obtained sequences had a high percentage similarity (98.36-99.62%) to A. platys sequences. The groEL sequences of three A. platys-like isolates were clustered in the same clade as the A. platys from the tick Rhipicephalus microplus in China.

CONCLUSIONS: Our data showed that the apparently healthy buffalo were naturally infected by bacteria in the family Anaplasmataceae at a relatively high prevalence. We also report the finding of A. platys-like infections in water buffalo in Thailand for the first time. Water buffalo serving as the reservoir host of anaplasmosis is of concern for managing the disease control and prevention in ruminants.}, } @article {pmid33027888, year = {2020}, author = {Bakovic, V and Schebeck, M and Stauffer, C and Schuler, H}, title = {Wolbachia-Mitochondrial DNA Associations in Transitional Populations of Rhagoletis cerasi.}, journal = {Insects}, volume = {11}, number = {10}, pages = {}, pmid = {33027888}, issn = {2075-4450}, support = {J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 31441/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {The endosymbiont Wolbachia can manipulate arthropod host reproduction by inducing cytoplasmic incompatibility (CI), which results in embryonic mortality when infected males mate with uninfected females. A CI-driven invasion of Wolbachia can result in a selective sweep of associated mitochondrial haplotype. The co-inheritance of Wolbachia and host mitochondrial DNA can therefore provide significant information on the dynamics of an ongoing Wolbachia invasion. Therefore, transition zones (i.e., regions where a Wolbachia strain is currently spreading from infected to uninfected populations) represent an ideal area to investigate the relationship between Wolbachia and host mitochondrial haplotype. Here, we studied Wolbachia-mitochondrial haplotype associations in the European cherry fruit fly, Rhagoletis cerasi, in two transition zones in the Czech Republic and Hungary, where the CI-inducing strain wCer2 is currently spreading. The wCer2-infection status of 881 individuals was compared with the two known R. cerasi mitochondrial haplotypes, HT1 and HT2. In accordance with previous studies, wCer2-uninfected individuals were associated with HT1, and wCer2-infected individuals were mainly associated with HT2. We found misassociations only within the transition zones, where HT2 flies were wCer2-uninfected, suggesting the occurrence of imperfect maternal transmission. We did not find any HT1 flies that were wCer2-infected, suggesting that Wolbachia was not acquired horizontally. Our study provides new insights into the dynamics of the early phase of a Wolbachia invasion.}, } @article {pmid33024036, year = {2020}, author = {Hague, MTJ and Caldwell, CN and Cooper, BS}, title = {Pervasive Effects of Wolbachia on Host Temperature Preference.}, journal = {mBio}, volume = {11}, number = {5}, pages = {}, pmid = {33024036}, issn = {2150-7511}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Behavior, Animal ; *Body Temperature ; Body Temperature Regulation ; Cytoplasm/microbiology ; Drosophila melanogaster/microbiology/*physiology ; Female ; Genotype ; Host Microbial Interactions/*physiology ; Male ; Phenotype ; Phylogeny ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Heritable symbionts can modify a range of ecologically important host traits, including behavior. About half of all insect species are infected with maternally transmitted Wolbachia, a bacterial endosymbiont known to alter host reproduction, nutrient acquisition, and virus susceptibility. Here, we broadly test the hypothesis that Wolbachia modifies host behavior by assessing the effects of eight different Wolbachia strains on the temperature preference of six Drosophila melanogaster subgroup species. Four of the seven host genotypes infected with A-group Wolbachia strains (wRi in Drosophila simulans, wHa in D. simulans, wSh in Drosophila sechellia, and wTei in Drosophila teissieri) prefer significantly cooler temperatures relative to uninfected genotypes. Contrastingly, when infected with divergent B-group wMau, Drosophila mauritiana prefers a warmer temperature. For most strains, changes to host temperature preference do not alter Wolbachia titer. However, males infected with wSh and wTei tend to experience an increase in titer when shifted to a cooler temperature for 24 h, suggesting that Wolbachia-induced changes to host behavior may promote bacterial replication. Our results indicate that Wolbachia modifications to host temperature preference are likely widespread, which has important implications for insect thermoregulation and physiology. Understanding the fitness consequences of these Wolbachia effects is crucial for predicting evolutionary outcomes of host-symbiont interactions, including how Wolbachia spreads to become common.IMPORTANCE Microbes infect a diversity of species, influencing the performance and fitness of their hosts. Maternally transmitted Wolbachia bacteria infect most insects and other arthropods, making these bacteria some of the most common endosymbionts in nature. Despite their global prevalence, it remains mostly unknown how Wolbachia influence host physiology and behavior to proliferate. We demonstrate pervasive effects of Wolbachia on Drosophila temperature preference. Most hosts infected with A-group Wolbachia prefer cooler temperatures, whereas the one host species infected with divergent B-group Wolbachia prefers warmer temperatures, relative to uninfected genotypes. Changes to host temperature preference generally do not alter Wolbachia abundance in host tissues, but for some A-group strains, adult males have increased Wolbachia titer when shifted to a cooler temperature. This suggests that Wolbachia-induced changes to host behavior may promote bacterial replication. Our results help elucidate the impact of endosymbionts on their hosts amid the global Wolbachia pandemic.}, } @article {pmid33023928, year = {2020}, author = {Lemon, A and Bordenstein, SR and Bordenstein, SR}, title = {Discover the Microbes Within! The Wolbachia Project: Citizen Science and Student-Based Discoveries for 15 Years and Counting.}, journal = {Genetics}, volume = {216}, number = {2}, pages = {263-268}, pmid = {33023928}, issn = {1943-2631}, mesh = {Animals ; Awards and Prizes ; Citizen Science/*methods/organization & administration ; Genetic Techniques ; Genetics/*education/organization & administration ; Humans ; *Microbiota ; Wolbachia/*genetics/pathogenicity ; }, abstract = {The Elizabeth W. Jones Award for Excellence in Education recognizes an individual who has had a significant impact on genetics education at any education level. Seth R. Bordenstein, Ph.D., Centennial Professor of Biological Sciences at Vanderbilt University and Founding Director of the Vanderbilt Microbiome Initiative, is the 2020 recipient in recognition of his cofounding, developing, and expanding Discover the Microbes Within! The Wolbachia Project.}, } @article {pmid33022249, year = {2020}, author = {Hoffmann, A}, title = {Wolbachia.}, journal = {Current biology : CB}, volume = {30}, number = {19}, pages = {R1113-R1114}, doi = {10.1016/j.cub.2020.08.039}, pmid = {33022249}, issn = {1879-0445}, mesh = {Animals ; *Bacterial Load ; *Host-Pathogen Interactions ; Invertebrates/*parasitology ; *Reproduction ; Wolbachia/*physiology ; }, abstract = {Ary Hoffman introduces Wolbachia.}, } @article {pmid33016318, year = {2020}, author = {Moussa, A and Passera, A and Sanna, F and Faccincani, M and Casati, P and Bianco, PA and Mori, N and Quaglino, F}, title = {Bacterial microbiota associated with insect vectors of grapevine Bois noir disease in relation to phytoplasma infection.}, journal = {FEMS microbiology ecology}, volume = {96}, number = {11}, pages = {}, doi = {10.1093/femsec/fiaa203}, pmid = {33016318}, issn = {1574-6941}, mesh = {Insect Vectors ; *Microbiota ; Phylogeny ; Phytoplasma Disease ; Plant Diseases ; *Vitis ; }, abstract = {Bois noir is a grapevine disease causing severe yield loss in vineyards worldwide. It is associated with 'Candidatus Phytoplasma solani', a phloem-limited bacterium transmitted by polyphagous insects. Due to its complex epidemiology, it is difficult to organize effective containment measures. This study aimed to describe the bacterial microbiota associated with 'Candidatus Phytoplasma solani' infected and non-infected insect hosts and vectors to investigate if phytoplasma presence can shape the microbiota. Alpha-diversity analysis showed a low microbiota diversity in these insects, in which few genera were highly abundant. Beta-diversity analysis revealed that the xylem- and phloem-feeding behavior influences the microbiota structure. Moreover, it highlighted that phytoplasma infection is associated with a restructuring of microbiota exclusively in Deltocephalinae insect vectors. Obtained data showed that 'Candidatus Phytoplasma solani' may have adverse effects on the endosymbionts Sulcia and Wolbachia, suggesting a possible fitness modification in the insects. The phytoplasma-antagonistic Dyella was not found in any of the examined insect species. The results indicate an interesting perspective regarding the microbial signatures associated with xylem- and phloem-feeding insects, and determinants that could be relevant to establish whether an insect species can be a vector or not, opening up new avenues for developing microbial resource management-based approaches.}, } @article {pmid33004433, year = {2020}, author = {Mathers, TC and Mugford, ST and Hogenhout, SA and Tripathi, L}, title = {Genome Sequence of the Banana Aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae) and Its Symbionts.}, journal = {G3 (Bethesda, Md.)}, volume = {10}, number = {12}, pages = {4315-4321}, pmid = {33004433}, issn = {2160-1836}, support = {BB/R01227X/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P012574/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; *Aphids/genetics ; *Babuvirus ; *Buchnera ; *Musa ; *Wolbachia ; }, abstract = {The banana aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae), is a major pest of cultivated bananas (Musa spp., order Zingiberales), primarily due to its role as a vector of Banana bunchy top virus (BBTV), the most severe viral disease of banana worldwide. Here, we generated a highly complete genome assembly of P. nigronervosa using a single PCR-free Illumina sequencing library. Using the same sequence data, we also generated complete genome assemblies of the P. nigronervosa symbiotic bacteria Buchnera aphidicola and Wolbachia To improve our initial assembly of P. nigronervosa we developed a k-mer based deduplication pipeline to remove genomic scaffolds derived from the assembly of haplotigs (allelic variants assembled as separate scaffolds). To demonstrate the usefulness of this pipeline, we applied it to the recently generated assembly of the aphid Myzus cerasi, reducing the duplication of conserved BUSCO genes by 25%. Phylogenomic analysis of P. nigronervosa, our improved M. cerasi assembly, and seven previously published aphid genomes, spanning three aphid tribes and two subfamilies, reveals that P. nigronervosa falls within the tribe Macrosiphini, but is an outgroup to other Macrosiphini sequenced so far. As such, the genomic resources reported here will be useful for understanding both the evolution of Macrosphini and for the study of P. nigronervosa. Furthermore, our approach using low cost, high-quality, Illumina short-reads to generate complete genome assemblies of understudied aphid species will help to fill in genomic black spots in the diverse aphid tree of life.}, } @article {pmid33002324, year = {2021}, author = {Izraeli, Y and Lalzar, M and Netanel, N and Mozes-Daube, N and Steinberg, S and Chiel, E and Zchori-Fein, E}, title = {Wolbachia influence on the fitness of Anagyrus vladimiri (Hymenoptera: Encyrtidae), a bio-control agent of mealybugs.}, journal = {Pest management science}, volume = {77}, number = {2}, pages = {1023-1034}, doi = {10.1002/ps.6117}, pmid = {33002324}, issn = {1526-4998}, support = {20-02-0092//Chief Scientist of Agriculture, Israel/ ; }, mesh = {Animals ; Female ; *Hymenoptera ; Multilocus Sequence Typing ; Reproduction ; Symbiosis ; *Wasps ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Like numerous other animals, biocontrol agents (BCAs) of arthropod pests carry various microorganisms that may have diverse effects on the biology of their eukaryote hosts. We postulated that it is possible to improve the efficacy of BCAs by manipulating the composition of their associated microbiota. The parasitoid wasp Anagyrus vladimiri (Hymenoptera: Encyrtidae) from a mass-rearing facility was chosen for testing this hypothesis.

RESULTS: High-throughput sequencing analysis indicated that fungal abundance in A. vladimiri was low and variable, whereas the bacterial community was dominated by the endosymbiont Wolbachia. Wolbachia was fixed in the mass-rearing population, whereas in field-collected A. vladimiri Wolbachia's prevalence was only approximately 20%. Identification of Wolbachia strains from the two populations by Multi Locus Sequence Typing, revealed two closely related but unique strains. A series of bioassays with the mass-rearing Wolbachia-fixed (W[+]) and a derived antibiotic-treated Wolbachia-free (W[-]) lines revealed that: (i) Wolbachia does not induce reproductive manipulations; (ii) W[-] females have higher fecundity when reared individually, but not when reared with conspecifics; (iii) W[+] females outcompete W[-] when they share hosts for oviposition; (iv) longevity and developmental time were similar in both lines.

CONCLUSIONS: The findings suggest that W[+] A. vladimiri have no clear fitness benefit under mass-rearing conditions and may be disadvantageous under lab-controlled conditions. In a broader view, the results suggest that augmentative biological control can benefit from manipulation of the microbiome of natural enemies.}, } @article {pmid32975515, year = {2020}, author = {Shropshire, JD and Leigh, B and Bordenstein, SR}, title = {Symbiont-mediated cytoplasmic incompatibility: what have we learned in 50 years?.}, journal = {eLife}, volume = {9}, number = {}, pages = {}, pmid = {32975515}, issn = {2050-084X}, support = {R01 AI143725/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; F32 AI140694/AI/NIAID NIH HHS/United States ; IOS1456778//National Science Foundation/International ; DGE144519//National Science Foundation/International ; }, mesh = {Animals ; Arthropods/genetics/*physiology ; *Cytoplasm/genetics/metabolism/microbiology/physiology ; Female ; Male ; *Reproduction/genetics/physiology ; Species Specificity ; Symbiosis/*physiology ; Ubiquitins/genetics/metabolism ; }, abstract = {Cytoplasmic incompatibility (CI) is the most common symbiont-induced reproductive manipulation. Specifically, symbiont-induced sperm modifications cause catastrophic mitotic defects in the fertilized embryo and ensuing lethality in crosses between symbiotic males and either aposymbiotic females or females harboring a different symbiont strain. However, if the female carries the same symbiont strain, then embryos develop properly, thereby imparting a relative fitness benefit to symbiont-transmitting mothers. Thus, CI drives maternally-transmitted bacteria to high frequencies in arthropods worldwide. In the past two decades, CI experienced a boom in interest due to its (i) deployment in worldwide efforts to curb mosquito-borne diseases, (ii) causation by bacteriophage genes, cifA and cifB, that modify sexual reproduction, and (iii) important impacts on arthropod speciation. This review serves as a gateway to experimental, conceptual, and quantitative themes of CI and outlines significant gaps in understanding CI's mechanism that are ripe for investigation from diverse subdisciplines in the life sciences.}, } @article {pmid32969551, year = {2020}, author = {Price, TAR and Windbichler, N and Unckless, RL and Sutter, A and Runge, JN and Ross, PA and Pomiankowski, A and Nuckolls, NL and Montchamp-Moreau, C and Mideo, N and Martin, OY and Manser, A and Legros, M and Larracuente, AM and Holman, L and Godwin, J and Gemmell, N and Courret, C and Buchman, A and Barrett, LG and Lindholm, AK}, title = {Resistance to natural and synthetic gene drive systems.}, journal = {Journal of evolutionary biology}, volume = {33}, number = {10}, pages = {1345-1360}, pmid = {32969551}, issn = {1420-9101}, support = {NIH R35GM119515/NH/NIH HHS/United States ; P300PA_177906/SNSF_/Swiss National Science Foundation/Switzerland ; P300PA_177830/SNSF_/Swiss National Science Foundation/Switzerland ; ZSEZ0_180460/SNSF_/Swiss National Science Foundation/Switzerland ; 31003A_160328//Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung/ ; R35 GM119515/GM/NIGMS NIH HHS/United States ; }, mesh = {*Biological Evolution ; *Gene Drive Technology ; *Selection, Genetic ; }, abstract = {Scientists are rapidly developing synthetic gene drive elements intended for release into natural populations. These are intended to control or eradicate disease vectors and pests, or to spread useful traits through wild populations for disease control or conservation purposes. However, a crucial problem for gene drives is the evolution of resistance against them, preventing their spread. Understanding the mechanisms by which populations might evolve resistance is essential for engineering effective gene drive systems. This review summarizes our current knowledge of drive resistance in both natural and synthetic gene drives. We explore how insights from naturally occurring and synthetic drive systems can be integrated to improve the design of gene drives, better predict the outcome of releases and understand genomic conflict in general.}, } @article {pmid32966874, year = {2020}, author = {Guo, Y and Khan, J and Zheng, XY and Wu, Y}, title = {Wolbachia increase germ cell mitosis to enhance the fecundity of Laodelphax striatellus.}, journal = {Insect biochemistry and molecular biology}, volume = {127}, number = {}, pages = {103471}, doi = {10.1016/j.ibmb.2020.103471}, pmid = {32966874}, issn = {1879-0240}, mesh = {Animals ; Female ; Fertility ; Hemiptera/microbiology/*physiology ; *Mitosis ; Ovum/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular bacteria that infect a wide range of invertebrates and have evolved various strategies to alter host reproduction for their own survival and dissemination. In small brown planthopper Laodelphax striatellus, Wolbachia-infected females lay more eggs than uninfected females. Our previous study has shown that Wolbachia are abundant in ovarian cells of L. striatellus and change the number of apoptotic nurse cells in a caspase-dependent manner to provide nutrition for oogenesis. The cellular and molecular bases of the Wolbachia-mediated alterations in L. striatellus oogenesis remain largely unknown. Here, we investigated whether germ cell mitosis, which has been implicated in determination of egg production rates, influences the interaction between fecundity and Wolbachia in L.striatellus. We used an anti-phospho-histone 3 (pH3) antibody to label and visualize mitotic cells. Microscopic observations indicated that the Wolbachia strain wStri increased the number of ovarioles that contained mitotic germ cells. The increased fecundity of Wolbachia-infected females was a result of mitosis of germ cells; the frequency of germ cell mitosis was much higher in infected females than in uninfected females. In addition, mitosis inhibition by Cdc20, CDK1, and CycB messenger RNA interference in Wolbachia-infected L. striatellus markedly decreased egg numbers. Live Wolbachia recolonization enhanced the egg production of uninfected L. striatellus by directly affecting mitosis regulators. Together, these data suggest that wStri might increase germ cell mitosis to enhance the fecundity of L. striatellus in a mitosis-regulating manner. Our findings establish a link between Wolbachia-induced mitosis and Wolbachia-mediated egg production effects.}, } @article {pmid32948809, year = {2020}, author = {Koh, C and Islam, MN and Ye, YH and Chotiwan, N and Graham, B and Belisle, JT and Kouremenos, KA and Dayalan, S and Tull, DL and Klatt, S and Perera, R and McGraw, EA}, title = {Dengue virus dominates lipid metabolism modulations in Wolbachia-coinfected Aedes aegypti.}, journal = {Communications biology}, volume = {3}, number = {1}, pages = {518}, pmid = {32948809}, issn = {2399-3642}, support = {R01 AI151166/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*metabolism/microbiology/pathogenicity/virology ; Animals ; Dengue/genetics/metabolism/microbiology/virology ; Dengue Virus/*genetics/metabolism/pathogenicity ; Humans ; Insect Vectors/genetics/microbiology/virology ; Lipid Metabolism/*genetics ; Pest Control, Biological ; Virus Replication/genetics ; Wolbachia/*genetics/metabolism/pathogenicity ; }, abstract = {Competition between viruses and Wolbachia for host lipids is a proposed mechanism of Wolbachia-mediated virus blocking in insects. Yet, the metabolomic interaction between virus and symbiont within the mosquito has not been clearly defined. We compare the lipid profiles of Aedes aegypti mosquitoes bearing mono- or dual-infections of the Wolbachia wMel strain and dengue virus serotype 3 (DENV3). We found metabolic signatures of infection-induced intracellular events but little evidence to support direct competition between Wolbachia and virus for host lipids. Lipid profiles of dual-infected mosquitoes resemble those of DENV3 mono-infected mosquitoes, suggesting virus-driven modulation dominates over that of Wolbachia. Interestingly, knockdown of key metabolic enzymes suggests cardiolipins are host factors for DENV3 and Wolbachia replication. These findings define the Wolbachia-DENV3 metabolic interaction as indirectly antagonistic, rather than directly competitive, and reveal new research avenues with respect to mosquito × virus interactions at the molecular level.}, } @article {pmid32945029, year = {2021}, author = {Xia, X and Peng, CW and Cui, JR and Jin, PY and Yang, K and Hong, XY}, title = {Wolbachia affects reproduction in the spider mite Tetranychus truncatus (Acari: Tetranychidae) by regulating chorion protein S38-like and Rop.}, journal = {Insect molecular biology}, volume = {30}, number = {1}, pages = {18-29}, doi = {10.1111/imb.12669}, pmid = {32945029}, issn = {1365-2583}, mesh = {Animals ; Egg Proteins/metabolism ; Female ; Fertility ; Gene Expression ; *Host-Parasite Interactions ; Oviposition ; RNA Interference ; RNA-Seq ; Reproduction ; Symbiosis ; Tetranychidae/genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia-induced reproductive regulation in hosts has been used to control pest populations, but little is known about the molecular mechanism underlying Wolbachia regulation of host genes. Here, reproductive regulation by Wolbachia in the spider mite Tetranychus truncatus was studied at the molecular level. Infection with Wolbachia resulted in decreasing oviposition and cytoplasmic incompatibility in T. truncatus. Further RNA-seq revealed genes regulated by Wolbachia in T. truncatus. Real-time quantitative polymerase chain reaction (qPCR) showed that genes, including chorion protein S38-like and Rop were down-regulated by Wolbachia. RNA interference (RNAi) of chorion protein S38-like and Rop in Wolbachia-uninfected T. truncatus decreased oviposition, which was consistent with Wolbachia-induced oviposition decrease. Interestingly, suppressing Rop in Wolbachia-infected T. truncatus led to increased Wolbachia titres in eggs; however, this did not occur after RNAi of chorion protein S38-like. This is the first study to show that chorion protein S38-like and Rop facilitate Wolbachia-mediated changes in T. truncatus fertility. In addition, RNAi of Rop turned the body colour of Wolbachia-uninfected T. truncatus black, which indicates that the role of Rop is not limited to the reproductive regulation of T. truncatus.}, } @article {pmid32944607, year = {2020}, author = {Arham, AF and Amin, L and Mustapa, MAC and Mahadi, Z and Arham, AF and Yaacob, M and Ibrahim, M and Norizan, NS}, title = {Perceived benefits and risks: A survey data set towards Wolbachia-infected Aedes Mosquitoes in Klang Valley.}, journal = {Data in brief}, volume = {32}, number = {}, pages = {106262}, pmid = {32944607}, issn = {2352-3409}, abstract = {Perceived Benefits and Risks: A survey data set towards Wolbachia-infected Aedes Mosquitoes in the Klang Valley, Malaysia. Introduction: The paper presents data collected using measures of perceived benefits, perceived risks, trust in key players, attitude towards nature versus material, attitude towards technology, religiosity, and attitude towards the Wolbachia-infected Aedes mosquitoes (WiAM) technique. The validated questionnaires were used to randomly survey targeted stakeholders in the Klang Valley, Malaysia, who had been asked to voluntarily participate in face-to-face interviews. Completed questionnaires were received from 399 respondents (adults above 18 years old) and comprised two stakeholder groups: scientists (n = 202), and the public (n = 197). The detailed findings serve numerous opportunities to examine the social acceptance of Wolbachia-infected Aedes mosquitoes, to ensure the development of policy and action plans, and to encourage further study by other researchers interested in the measures and data presented.}, } @article {pmid32937243, year = {2020}, author = {Shin, PT and Baptista, RP and O'Neill, CM and Wallis, C and Reaves, BJ and Wolstenholme, AJ}, title = {Comparative sequences of the Wolbachia genomes of drug-sensitive and resistant isolates of Dirofilaria immitis.}, journal = {Veterinary parasitology}, volume = {286}, number = {}, pages = {109225}, pmid = {32937243}, issn = {1873-2550}, support = {R01 AI103140/AI/NIAID NIH HHS/United States ; R21 AI125899/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Dirofilaria immitis/*drug effects/microbiology ; *Drug Resistance ; *Genetic Variation ; *Genome, Bacterial ; Lactones/*pharmacology ; Macrocyclic Compounds/pharmacology ; Wolbachia/*genetics ; }, abstract = {The recent identification of isolates of D. immitis with confirmed resistance to the macrocyclic lactone preventatives presents an opportunity for comparative genomic studies using these isolates, and examining the genetic diversity within and between them. We studied the genomes of Wolbachia endosymbionts of five isolates of D. immitis maintained at the University of Georgia. Missouri and Georgia-2 are maintained as drug susceptible isolates, and JYD-27, Yazoo-2013 and Metairie-2014 are resistant to the macrocyclic lactone preventatives. We used whole genome amplification followed by Illumina-based sequencing from 8 to 12 individual microfilariae from each of the five isolates, obtaining a depth of coverage of approximately 40-75 fold for each. The Illumina sequences were used to create new genome assemblies for all the Wolbachia isolates studied. Comparisons of the Wolbachia sequences revealed more than 3000 sequence variations in each isolate. We identified 67 loci specific in resistant isolates but not in susceptible isolates, including 18 genes affected.Phylogenetic analysis suggested that the endosymbionts of the drug-susceptible isolates are more closely related to each other than to those from any of the resistant parasites. This level of variation in the Wolbachia endosymbionts of D. immitis isolates suggests a potential for selection for resistance against drugs targeting them.}, } @article {pmid32932887, year = {2020}, author = {Ge, C and Hu, J and Zhao, Z and Hoffmann, AA and Ma, S and Shen, L and Fang, J and Zhu, J and Yu, W and Jiang, W}, title = {Phylogeny and Density Dynamics of Wolbachia Infection of the Health Pest Paederus fuscipes Curtis (Coleoptera: Staphylinidae).}, journal = {Insects}, volume = {11}, number = {9}, pages = {}, pmid = {32932887}, issn = {2075-4450}, support = {31401997//National Natural Science Foundation of China/ ; 2019112//Shanghai Municipal Human Resources and Social Security Bureau/ ; }, abstract = {The maternally inherited obligate intracellular bacteria Wolbachia infects the reproductive tissues of a wide range of arthropods and affects host reproduction. Wolbachia is a credible biocontrol agent for reducing the impact of diseases associated with arthropod vectors. Paederus fuscipes is a small staphylinid beetle that causes dermatitis linearis and conjunctivitis in humans when they come into contact with skin. Wolbachia occur in this beetle, but their relatedness to other Wolbachia, their infection dynamics, and their potential host effects remain unknown. In this study, we report the phylogenetic position and density dynamics of Wolbachia in P. fuscipes. The phylogeny of Wolbachia based on an analysis of MLST genotyping showed that the bacteria from P. fuscipes belong to supergroup B. Quantitative PCR indicated that the infection density in adults was higher than in any other life stage (egg, larva or pupa), and that reproductive tissue in adults had the highest infection densities, with similar densities in the sexes. These findings provide a starting point for understanding the Wolbachia infection dynamics in P. fuscipes, and interactions with other components of the microbiota.}, } @article {pmid32927527, year = {2020}, author = {Kakumanu, ML and DeVries, ZC and Barbarin, AM and Santangelo, RG and Schal, C}, title = {Bed bugs shape the indoor microbial community composition of infested homes.}, journal = {The Science of the total environment}, volume = {743}, number = {}, pages = {140704}, doi = {10.1016/j.scitotenv.2020.140704}, pmid = {32927527}, issn = {1879-1026}, mesh = {Aged ; Animals ; *Bedbugs ; Hot Temperature ; Housing ; Humans ; *Microbiota ; Poverty ; }, abstract = {Indoor pests, and the allergens they produce, adversely affect human health. Surprisingly, however, their effects on indoor microbial communities have not been assessed. Bed bug (Cimex lectularius) infestations pose severe challenges in elderly and low-income housing. They void large amounts of liquid feces into the home environment, which might alter the indoor microbial community composition. In this study, using bed bug-infested and uninfested homes, we showed a strong impact of bed bug infestations on the indoor microbial diversity. Floor dust samples were collected from uninfested and bed bug-infested homes and their microbiomes were analyzed before and after heat interventions that eliminated bed bugs. The microbial communities of bed bug-infested homes were radically different from those of uninfested homes, and the bed bug endosymbiont Wolbachia was the major driver of this difference. After bed bugs were eliminated, the microbial community gradually shifted toward the community composition of uninfested homes, strongly implicating bed bugs in shaping the dust-associated environmental microbiome. Further studies are needed to understand the viability of these microbial communities and the potential risks that bed bug-associated microbes and their metabolites pose to human health.}, } @article {pmid32913052, year = {2020}, author = {Manokaran, G and Flores, HA and Dickson, CT and Narayana, VK and Kanojia, K and Dayalan, S and Tull, D and McConville, MJ and Mackenzie, JM and Simmons, CP}, title = {Modulation of acyl-carnitines, the broad mechanism behind Wolbachia-mediated inhibition of medically important flaviviruses in Aedes aegypti.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {117}, number = {39}, pages = {24475-24483}, pmid = {32913052}, issn = {1091-6490}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/chemistry/metabolism/*microbiology/*virology ; Animals ; Carnitine/chemistry/*metabolism ; Female ; Mosquito Vectors/chemistry/metabolism/microbiology/virology ; Wolbachia/*physiology ; Zika Virus/*physiology ; }, abstract = {Wolbachia-infected mosquitoes are refractory to flavivirus infections, but the role of lipids in Wolbachia-mediated virus blocking remains to be elucidated. Here, we use liquid chromatography mass spectrometry to provide a comprehensive picture of the lipidome of Aedes aegypti (Aag2) cells infected with Wolbachia only, either dengue or Zika virus only, and Wolbachia-infected Aag2 cells superinfected with either dengue or Zika virus. This approach identifies a class of lipids, acyl-carnitines, as being down-regulated during Wolbachia infection. Furthermore, treatment with an acyl-carnitine inhibitor assigns a crucial role for acyl-carnitines in the replication of dengue and Zika viruses. In contrast, depletion of acyl-carnitines increases Wolbachia density while addition of commercially available acyl-carnitines impairs Wolbachia production. Finally, we show an increase in flavivirus infection of Wolbachia-infected cells with the addition of acyl-carnitines. This study uncovers a previously unknown role for acyl-carnitines in this tripartite interaction that suggests an important and broad mechanism that underpins Wolbachia-mediated pathogen blocking.}, } @article {pmid32905752, year = {2020}, author = {Caragata, EP and Dong, S and Dong, Y and Simões, ML and Tikhe, CV and Dimopoulos, G}, title = {Prospects and Pitfalls: Next-Generation Tools to Control Mosquito-Transmitted Disease.}, journal = {Annual review of microbiology}, volume = {74}, number = {}, pages = {455-475}, doi = {10.1146/annurev-micro-011320-025557}, pmid = {32905752}, issn = {1545-3251}, mesh = {Animals ; *Biological Control Agents ; *Communicable Disease Control ; Communicable Diseases/*parasitology/transmission/*virology ; Culicidae/*genetics/parasitology/physiology/virology ; Humans ; Infertility ; Malaria ; Mosquito Control/*methods ; Wolbachia/genetics ; }, abstract = {Mosquito-transmitted diseases, including malaria and dengue, are a major threat to human health around the globe, affecting millions each year. A diverse array of next-generation tools has been designed to eliminate mosquito populations or to replace them with mosquitoes that are less capable of transmitting key pathogens. Many of these new approaches have been built on recent advances in CRISPR/Cas9-based genome editing. These initiatives have driven the development of pathogen-resistant lines, new genetics-based sexing methods, and new methods of driving desirable genetic traits into mosquito populations. Many other emerging tools involve microorganisms, including two strategies involving Wolbachia that are achieving great success in the field. At the same time, other mosquito-associated bacteria, fungi, and even viruses represent untapped sources of new mosquitocidal or antipathogen compounds. Although there are still hurdles to be overcome, the prospect that such approaches will reduce the impact of these diseases is highly encouraging.}, } @article {pmid32904796, year = {2020}, author = {Ciuca, L and Vismarra, A and Lebon, W and Beugnet, F and Morchon, R and Rinaldi, L and Cringoli, G and Kramer, L and Genchi, M}, title = {New insights into the biology, diagnosis and immune response to Dirofilaria repens in the canine host.}, journal = {Veterinary parasitology: X}, volume = {4}, number = {}, pages = {100029}, pmid = {32904796}, issn = {2590-1389}, abstract = {Dogs are the primary host for Dirofilaria repens, therefore it is mandatory to accurately diagnose the canine infection and to expand our current knowledge on parasite biology and the immune response of the infected host for a better prevention.Thus, the aim of the present study was to provide new insights from experimental infections of dogs with D. repens, focusing on the evaluation of: 1) the pre-patent period and 2) the antibody response against D. repens somatic antigens and against the Wolbachia endosymbiont. Briefly, on Day 0, twenty purpose-bred Beagle dogs were experimentally infected with 50 infective larvae (L3) of D. repens. Starting from Day 58 until the last day of the study (Day 281), blood samples were collected on a monthly basis for detection of antibodies against D. repens (Dr) and recombinant Wolbachia surface protein (rWSP) by non-commercial IgG-ELISAs. Additional samples were collected on Days 220, 245 and 281 for the detection of microfilariae (mff) using the modified Knott's test and biomolecular analysis, following two PCR protocols: Gioia et al. (2010; protocol A) and Rishniw et al. (2006- protocol B). The results were analysed by univariate statistical analyses using 2 × 2 contingency tables and K Cohen was calculated to assess the agreement among all the diagnostic techniques. Overall, the outcome of the study revealed that out of the 20 dogs experimentally infected with D. repens, 16 (80 %) were microfilaraemic, 17 (85 %) were positive at DNA detection in the blood, 18 (90 %) had D. repens antibodies and 16 (80 %) had Wolbachia antibodies on the last day of the study. The overall k agreement between Knott's and PCR protocol B was 0.442 (P = 0.0001) and increased throughout the study, reaching 0.828 (P = 0.0001) on Day 281. To the authors knowledge, this is only the second study reporting antibody response to D. repens somatic antigen in experimentally infected dogs. ELISA results showed that an antibody response develops before the onset of patency, and steadily increases with time. Results would suggest that the development of an immunological response to infection could lead to application in epidemiological studies, risk assessment and as an aid in the diagnostic approach in dogs, in particular for early infections without mff.}, } @article {pmid32888935, year = {2020}, author = {Lee, JM and Yek, SH and Wilson, RF and Rahman, S}, title = {Characterization of the Aedes albopictus (Diptera: Culicidae) holobiome: bacterial composition across land use type and mosquito sex in Malaysia.}, journal = {Acta tropica}, volume = {212}, number = {}, pages = {105683}, doi = {10.1016/j.actatropica.2020.105683}, pmid = {32888935}, issn = {1873-6254}, mesh = {Aedes/*microbiology ; Animals ; Bacteria/*isolation & purification ; Female ; Malaysia ; Male ; Microbial Interactions ; *Microbiota ; Wolbachia/isolation & purification ; }, abstract = {Understanding the diversity and dynamics of the microbiota within the mosquito holobiome is of great importance to apprehend how the microbiota modulates various complex processes and interactions. This study examined the bacterial composition of Aedes albopictus across land use type and mosquito sex in the state of Selangor, Malaysia using 16S rRNA sequencing. The bacterial community structure in mosquitoes was found to be influenced by land use type and mosquito sex, with the environment and mosquito diet respectively identified to be the most likely sources of microbes. We found that approximately 70% of the microbiota samples were dominated by Wolbachia and removing Wolbachia from analyses revealed the relatively even composition of the remaining bacterial microbiota. Furthermore, microbial interaction network analysis highlighted the prevalence of co-exclusionary patterns in all networks regardless of land use and mosquito sex, with Wolbachia exhibiting co-exclusionary interactions with other residential bacteria such as Xanthomonas, Xenophilus and Zymobacter.}, } @article {pmid32884660, year = {2020}, author = {Noh, P and Oh, SY and Park, S and Kwon, T and Kim, Y and Choe, JC and Jeong, G}, title = {Association between host wing morphology polymorphism and Wolbachia infection in Vollenhovia emeryi (Hymenoptera: Myrmicinae).}, journal = {Ecology and evolution}, volume = {10}, number = {16}, pages = {8827-8837}, pmid = {32884660}, issn = {2045-7758}, abstract = {Many eusocial insects, including ants, show complex colony structures, distributions, and reproductive strategies. In the ant Vollenhovia emeryi Wheeler (Hymenoptera: Myrmicinae), queens and males are produced clonally, while sterile workers arise sexually, unlike other ant species and Hymenopteran insects in general. Furthermore, there is a wing length polymorphism in the queen caste. Despite its evolutionary remarkable traits, little is known about the population structure of this ant species, which may provide insight into its unique reproductive mode and polymorphic traits. We performed in-depth analyses of ant populations from Korea, Japan, and North America using three mitochondrial genes (COI, COII, and Cytb). The long-winged (L) morph is predominant in Korean populations, and the short-winged (S) morph is very rare. Interestingly, all L morphs were infected with Wolbachia, while all Korean S morphs lacked Wolbachia, demonstrating a association between a symbiont and a phenotypic trait. A phylogenetic analysis revealed that the S morph is derived from the L morph. We propose that the S morph is associated with potential resistance to Wolbachia infection and that Wolbachia infection does not influence clonal reproduction (as is the case in other ant species).}, } @article {pmid32881871, year = {2020}, author = {Zhang, D and Xi, Z and Li, Y and Wang, X and Yamada, H and Qiu, J and Liang, Y and Zhang, M and Wu, Y and Zheng, X}, title = {Toward implementation of combined incompatible and sterile insect techniques for mosquito control: Optimized chilling conditions for handling Aedes albopictus male adults prior to release.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {9}, pages = {e0008561}, pmid = {32881871}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Cold Temperature ; Female ; Male ; Mosquito Control/*methods ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Combined incompatible and sterile insect technique (IIT-SIT) has been considered to be an effective and safe approach to control mosquito populations. Immobilization of male adults by chilling is a crucial process required for the packing, transportation and release of the mosquitoes during the implementation of IIT-SIT for mosquito control. In this study, effects of chilling on the Aedes albopictus males with triple Wolbachia infections (HC line), a powerful weapon to fight against the wild type Ae. albopictus population via IIT-SIT, were evaluated under both laboratory and field conditions. Irradiated HC (IHC) males were exposed to 1, 5 and 10°C for 1, 2, 3, 6 and 24 h. The survival rate of the post-chilled IHC males was then monitored. Longevity of post-chilled IHC males was compared to non-chilled males under laboratory and semi-field conditions. Mating competitiveness of IHC/HC males after exposure to 5 or 10°C for 0, 3 and 24 h was then evaluated. Effects of compaction and transportation under chilled conditions on the survival rate of IHC males were also monitored. The optimal chilling conditions for handling IHC males were temperatures between 5 and 10°C for a duration of less than 3 h with no negative impacts on survival rate, longevity and mating competitiveness when compared to non-chilled males. However, the overall quality of post-chilled IHC/HC males decreased when exposed to low temperatures for 24 h. Reduced survival was observed when IHC males were stored at 5°C under a compaction height of 8 cm. Transportation with chilling temperatures fluctuating from 8 to 12°C has no negative impact on the survival of IHC males. This study identified the optimal chilling temperature and duration for the handling and transportation of Ae. albopictus IHC male adults without any detrimental effect on their survival, longevity and mating competitiveness. Further studies are required to develop drone release systems specific for chilled mosquitoes to improve release efficiency, as well as to compare the population suppression efficiency between release of post-chilled and non-chilled males in the field.}, } @article {pmid32878094, year = {2020}, author = {Castillo, AM and Saltonstall, K and Arias, CF and Chavarria, KA and Ramírez-Camejo, LA and Mejía, LC and De León, LF}, title = {The Microbiome of Neotropical Water Striders and Its Potential Role in Codiversification.}, journal = {Insects}, volume = {11}, number = {9}, pages = {}, pmid = {32878094}, issn = {2075-4450}, support = {No. 270-2013-284//Secretaría Nacional de Ciencia, Tecnología e Innovación/ ; No. FID16-116//Secretaría Nacional de Ciencia, Tecnología e Innovación/ ; 152-2018//Sistema Nacional de Investigación, SNI/ ; }, abstract = {Insects host a highly diverse microbiome, which plays a crucial role in insect life. However, the composition and diversity of microbiomes associated with Neotropical freshwater insects is virtually unknown. In addition, the extent to which diversification of this microbiome is associated with host phylogenetic divergence remains to be determined. Here, we present the first comprehensive analysis of bacterial communities associated with six closely related species of Neotropical water striders in Panama. We used comparative phylogenetic analyses to assess associations between dominant bacterial linages and phylogenetic divergence among species of water striders. We found a total of 806 16S rRNA amplicon sequence variants (ASVs), with dominant bacterial taxa belonging to the phyla Proteobacteria (76.87%) and Tenericutes (19.51%). Members of the α- (e.g., Wolbachia) and γ- (e.g., Acinetobacter, Serratia) Proteobacteria, and Mollicutes (e.g., Spiroplasma) were predominantly shared across species, suggesting the presence of a core microbiome in water striders. However, some bacterial lineages (e.g., Fructobacillus, Fluviicola and Chryseobacterium) were uniquely associated with different water strider species, likely representing a distinctive feature of each species' microbiome. These findings indicate that both host identity and environmental context are important drivers of microbiome diversity in water striders. In addition, they suggest that diversification of the microbiome is associated with diversification in water striders. Although more research is needed to establish the evolutionary consequences of host-microbiome interaction in water striders, our findings support recent work highlighting the role of bacterial community host-microbiome codiversification.}, } @article {pmid32857984, year = {2021}, author = {Brattig, NW and Cheke, RA and Garms, R}, title = {Onchocerciasis (river blindness) - more than a century of research and control.}, journal = {Acta tropica}, volume = {218}, number = {}, pages = {105677}, doi = {10.1016/j.actatropica.2020.105677}, pmid = {32857984}, issn = {1873-6254}, mesh = {Animals ; Humans ; Insect Vectors/parasitology ; Onchocerciasis, Ocular/epidemiology/*prevention & control/transmission ; Prevalence ; }, abstract = {This review summarises more than a century of research on onchocerciasis, also known as river blindness, and its control. River blindness is an infection caused by the tissue filaria Onchocerca volvulus affecting the skin, subcutaneous tissue and eyes and leading to blindness in a minority of infected persons. The parasite is transmitted by its intermediate hosts Simulium spp. which breed in rivers. Featured are history and milestones in onchocerciasis research and control, state-of-the-art data on the parasite, its endobacteria Wolbachia, on the vectors, previous and current prevalence of the infection, its diagnostics, the interaction between the parasite and its host, immune responses and the pathology of onchocerciasis. Detailed information is documented on the time course of control programmes in the afflicted countries in Africa and the Americas, a long road from previous programmes to current successes in control of the transmission of this infectious disease. By development, adjustment and optimization of the control measures, transmission by the vector has been interrupted in foci of countries in the Americas, in Uganda, in Sudan and elsewhere, followed by onchocerciasis eliminations. The current state and future perspectives for control, elimination and eradication within the next 20-30 years are described and discussed. This review contributes to a deeper comprehension of this disease by a tissue-dwelling filaria and it will be helpful in efforts to control and eliminate other filarial infections.}, } @article {pmid32855244, year = {2020}, author = {Neupane, S and Bonilla, SI and Manalo, AM and Pelz-Stelinski, KS}, title = {Near-Complete Genome Sequences of a Wolbachia Strain Isolated from Diaphorina citri Kuwayama (Hemiptera: Liviidae).}, journal = {Microbiology resource announcements}, volume = {9}, number = {35}, pages = {}, pmid = {32855244}, issn = {2576-098X}, abstract = {Wolbachia strains are one of three endosymbionts associated with the insect vector of "Candidatus Liberibacter asiaticus," Diaphorina citri Kuwayama (Hemiptera: Liviidae). We report three near-complete genome sequences of samples of Wolbachia from D. citri (wDi), with sizes of 1,518,595, 1,542,468, and 1,538,523 bp.}, } @article {pmid32854447, year = {2020}, author = {Adegoke, A and Kumar, D and Bobo, C and Rashid, MI and Durrani, AZ and Sajid, MS and Karim, S}, title = {Tick-Borne Pathogens Shape the Native Microbiome Within Tick Vectors.}, journal = {Microorganisms}, volume = {8}, number = {9}, pages = {}, pmid = {32854447}, issn = {2076-2607}, support = {P20 GM103476/GM/NIGMS NIH HHS/United States ; }, abstract = {Ticks are blood-feeding arthropods and transmit a variety of medically important viral, bacterial, protozoan pathogens to animals and humans. Ticks also harbor a diverse community of microbes linked to their biological processes, such as hematophagy, and hence affect vector competence. The interactions between bacterial and/or protozoan pathogens and the tick microbiome is a black-box, and therefore we tested the hypothesis that the presence of a protozoan or bacterial pathogen will alter the microbial composition within a tick. Hence, this study was designed to define the microbial composition of two tick species, Hyalomma (H.) anatolicum and Rhipicephalus (R.) microplus. We used a combination of PCR based pathogen (Anaplasma marginale and Theileria species) and symbiont (Wolbachia species) identification followed by metagenomic sequencing and comparison of the microbial communities in PCR positive and negative ticks. A total of 1786 operational taxonomic units was identified representing 25 phyla, 50 classes, and 342 genera. The phylum Proteobacteria, Firmicutes, Actinobacteriota, and Bacteroidota were the most represented bacteria group. Alpha and beta diversity were not significantly affected in the presence or absence of Theileria sp. and A. marginale as see with H. anatolicum ticks. Interestingly, bacterial communities were significantly reduced in Theileria sp. infected R. microplus ticks, while also exhibiting a significant reduction in microbial richness and evenness. Putting these observations together, we referred to the effect the presence of Theileria sp. has on R. microplus a "pathogen-induced dysbiosis". We also identify the presence of Plasmodium falciparum, the causative agent of human malaria from the microbiome of both H. anatolicum and R. microplus ticks. These findings support the presence of a "pathogen-induced dysbiosis" within the tick and further validation experiments are required to investigate how they are important in the vector competence of ticks. Understanding the mechanism of "pathogen-induced dysbiosis" on tick microbial composition may aid the discovery of intervention strategies for the control of emerging tick-borne infections.}, } @article {pmid32854141, year = {2020}, author = {Altinli, M and Lequime, S and Atyame, C and Justy, F and Weill, M and Sicard, M}, title = {Wolbachia modulates prevalence and viral load of Culex pipiens densoviruses in natural populations.}, journal = {Molecular ecology}, volume = {29}, number = {20}, pages = {4000-4013}, doi = {10.1111/mec.15609}, pmid = {32854141}, issn = {1365-294X}, mesh = {Animals ; *Culex/genetics ; *Densovirus/genetics ; Phylogeny ; Prevalence ; Viral Load ; *Wolbachia/genetics ; }, abstract = {The inadequacy of standard mosquito control strategies calls for ecologically safe novel approaches, for example the use of biological agents such as the endosymbiotic α-proteobacteria Wolbachia or insect-specific viruses (ISVs). Understanding the ecological interactions between these "biocontrol endosymbionts" is thus a fundamental step. Wolbachia are transmitted vertically from mother to offspring and modify their hosts' phenotypes, including reproduction (e.g., cytoplasmic incompatibility) and survival (e.g., viral interference). In nature, Culex pipiens (sensu lato) mosquitoes are always found infected with genetically diverse Wolbachia called wPip that belong to five phylogenetic groups. In recent years, ISVs have also been discovered in these mosquito species, although their interactions with Wolbachia in nature are unknown. Here, we studied the interactions between a widely prevalent ISV, the Culex pipiens densovirus (CpDV, Densovirinae), and Wolbachia in northern Tunisian C. pipiens populations. We showed an influence of different Wolbachia groups on CpDV prevalence and a general positive correlation between Wolbachia and CpDV loads. By investigating the putative relationship between CpDV diversification and wPip groups in the different sites, we detected a signal linked to wPip groups in CpDV phylogeny in sites where all larvae were infected by the same wPip group. However, no such signal was detected where the wPip groups coexisted, suggesting CpDV horizontal transfer between hosts. Overall, our results provide good evidence for an ecological influence of Wolbachia on an ISV, CpDV, in natural populations and highlight the importance of integrating Wolbachia in our understanding of ISV ecology in nature.}, } @article {pmid32853197, year = {2020}, author = {Sim, S and Ng, LC and Lindsay, SW and Wilson, AL}, title = {A greener vision for vector control: The example of the Singapore dengue control programme.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {8}, pages = {e0008428}, pmid = {32853197}, issn = {1935-2735}, support = {/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; MR/P027873/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/virology ; Animals ; Community Participation ; Dengue/epidemiology/*prevention & control ; Humans ; Mosquito Control/*methods/*organization & administration ; Mosquito Vectors/virology ; Singapore/epidemiology ; }, abstract = {Vector-borne diseases are a major cause of morbidity and mortality worldwide. Aedes-borne diseases, in particular, including dengue, chikungunya, yellow fever, and Zika, are increasing at an alarming rate due to urbanisation, population movement, weak vector control programmes, and climate change. The World Health Organization calls for strengthening of vector control programmes in line with the Global Vector Control Response (GVCR) strategy, and many vector control programmes are transitioning to this new approach. The Singapore dengue control programme, situated within the country's larger vision of a clean, green, and sustainable environment for the health and well-being of its citizens, provides an excellent example of the GVCR approach in action. Since establishing vector control operations in the 1960s, the Singapore dengue control programme succeeded in reducing the dengue force of infection 10-fold by the 1990s and has maintained it at low levels ever since. Key to this success is consideration of dengue as an environmental disease, with a strong focus on source reduction and other environmental management methods as the dominant vector control strategy. The programme collaborates closely with other government ministries, as well as town councils, communities, the private sector, and academic and research institutions. Community engagement programmes encourage source reduction, and house-to-house inspections accompanied by a strong legislative framework with monetary penalties help to support compliance. Strong vector and epidemiological surveillance means that routine control activities can be heightened to specifically target dengue clusters. Despite its success, the programme continues to innovate to tackle challenges such as climate change, low herd immunity, and manpower constraints. Initiatives include development of novel vector controls such as Wolbachia-infected males and spatiotemporal models for dengue risk assessment. Lessons learnt from the Singapore programme can be applied to other settings, even those less well-resourced than Singapore, for more effective vector control.}, } @article {pmid32852571, year = {2021}, author = {Hubert, J and Nesvorna, M and Green, SJ and Klimov, PB}, title = {Microbial Communities of Stored Product Mites: Variation by Species and Population.}, journal = {Microbial ecology}, volume = {81}, number = {2}, pages = {506-522}, pmid = {32852571}, issn = {1432-184X}, support = {GA19-09998S//Grantová Agentura České Republiky/ ; 19-14-00004//Russian Science Foundation/ ; }, mesh = {Acaridae/classification/growth & development/*microbiology ; Animals ; Bacteria/classification/genetics/isolation & purification ; Diet ; Feces/microbiology ; Host Microbial Interactions ; *Microbiota ; Ovum/microbiology ; Phylogeny ; }, abstract = {Arthropod-associated microorganisms are important because they affect host fitness, protect hosts from pathogens, and influence the host's ability to vector pathogens. Stored product mites (Astigmata) often establish large populations in various types of food items, damaging the food by direct feeding and introducing contaminants, including their own bodies, allergen-containing feces, and associated microorganisms. Here we access the microbial structure and abundance in rearing diets, eggs, feces fraction, and mite bodies of 16 mite populations belonging to three species (Carpoglyphus lactis, Acarus siro, and Tyrophagus putrescentiae) using quantitative PCR and 16S ribosomal RNA (rRNA) gene amplicon sequencing. The mite microbiomes had a complex structure dominated by the following bacterial taxa (OTUs): (a) intracellular symbionts of the genera Cardinium and Wolbachia in the mite bodies and eggs; (b) putative gut symbionts of the genera Solitalea, Bartonella, and Sodalis abundant in mite bodies and also present in mite feces; (c) feces-associated or environmental bacteria of the genera Bacillus, Staphylococcus, and Kocuria in the diet, mite bodies, and feces. Interestingly and counterintuitively, the differences between microbial communities in various conspecific mite populations were higher than those between different mite species. To explain some of these differences, we hypothesize that the intracellular bacterial symbionts can affect microbiome composition in mite bodies, causing differences between microbial profiles. Microbial profiles differed between various sample types, such as mite eggs, bodies, and the environment (spent growth medium-SPGM). Low bacterial abundances in eggs may result in stochastic effects in parent-offspring microbial transmission, except for the intracellular symbionts. Bacteria in the rearing diet had little effect on the microbial community structure in SPGM and mite bodies. Mite fitness was positively correlated with bacterial abundance in SPGM and negatively correlated with bacterial abundances in mite bodies. Our study demonstrates critical host-microbe interactions, affecting all stages of mite growth and leading to alteration of the environmental microbiome. Correlational evidence based on absolute quantitation of bacterial 16S rRNA gene copies suggests that mite-associated microorganisms are critical for modulating important pest properties of mites by altering population growth.}, } @article {pmid32851048, year = {2020}, author = {Esteban-Mendoza, MV and Arcila-Quiceno, V and Albarracín-Navas, J and Hernández, I and Flechas-Alarcón, MC and Morchón, R}, title = {Current Situation of the Presence of Dirofilaria immitis in Dogs and Humans in Bucaramanga, Colombia.}, journal = {Frontiers in veterinary science}, volume = {7}, number = {}, pages = {488}, pmid = {32851048}, issn = {2297-1769}, abstract = {The cardiopulmonary dirofilariosis caused by Dirofilaria immitis, is a vector-borne infection, which can be transmitted to humans. The main hosts are both domestic and wild canids. This species mainly occurs in tropical and subtropical climates, and temperature and humidity are the main factors that favor the presence and proliferation of culicid mosquitoes as vectors of the disease. There are few reports of cardiopulmonary dirofilariosis in dogs and humans in Colombia, a region with favorable climatic conditions which favors the presence of mosquitoes that act as vectors of the disease. Therefore, this study aimed to examine its current prevalence in dogs and the risk of human exposure to the disease in Bucaramanga, one of the most populated areas in Colombia located at the center of the country. Furthermore, its demographic and environmental characteristics could be useful as a study model for other similar locations and neighboring countries. Serum samples from 351 dogs and 506 humans from the Bucaramanga Metropolitan area were analyzed. All dog samples were analyzed by Knott's technique and tested with a commercial immunochromatographic to detect the presence of circulating antigens of D. immitis. Human samples were analyzed using a non-commercial ELISA test kit to detect IgG against the somatic antigens of adult D. immitis and Wolbachia. Positive results were further confirmed using western blot analysis. Thirty-eight dogs tested positive with a overall prevalence of 10.82%. Of these dogs, 18 showed D. immitis microfilariae, being 5.12% of the total population. The overall seroprevalence in humans was 6.71%; seroprevalence was significantly higher in individuals aged 16-34 years-old and in women than in men. To our knowledge, this study describes seropositivity to D. immitis for the first time in a Colombian human population located in the same area as that of dogs infected with D. immitis, which represents a potential threat to public health. In humans, age and gender can be considered risk factors for exposure to D. immitis.}, } @article {pmid32849379, year = {2020}, author = {Lu, P and Sun, Q and Fu, P and Li, K and Liang, X and Xi, Z}, title = {Wolbachia Inhibits Binding of Dengue and Zika Viruses to Mosquito Cells.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {1750}, pmid = {32849379}, issn = {1664-302X}, abstract = {As traditional approaches to the control of dengue and Zika are insufficient, significant efforts have been made to develop utilization of the endosymbiotic bacterium Wolbachia to reduce the ability of mosquitoes to transmit pathogens. Although Wolbachia is known to inhibit flaviviruses in mosquitoes, including dengue virus (DENV) and Zika virus (ZIKV), it remains unclear how the endosymbiont interferes with viral replication cycle. In this study, we have carried out viral binding assays to investigate the impact of the Wolbachia strain wAlbB on the attachment of DENV serotype 2 (DENV-2) and ZIKV to Aedes aegypti Aag-2 cells. RNA interference (RNAi) was used to silence a variety of putative mosquito receptors of DENV that were differentially regulated by wAlbB in Aag-2 cells, in order to identify host factors involved in the inhibition of viral binding. Our results showed that, in addition to suppression of viral replication, Wolbachia strongly inhibited binding of both DENV-2 and ZIKV to Aag-2 cells. Moreover, the expression of two putative mosquito DENV receptors - dystroglycan and tubulin - was downregulated by wAlbB, and their knock-down resulted in the inhibition of DENV-2 binding to Aag-2 cells. These results will aid in understanding the Wolbachia-DENV interactions in mosquito and the development of novel control strategies for mosquito-borne diseases.}, } @article {pmid32831122, year = {2020}, author = {Wuliandari, JR and Hoffmann, AA and Tantowijoyo, W and Endersby-Harshman, NM}, title = {Frequency of kdr mutations in the voltage-sensitive sodium channel (VSSC) gene in Aedes aegypti from Yogyakarta and implications for Wolbachia-infected mosquito trials.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {429}, pmid = {32831122}, issn = {1756-3305}, support = {1037003//National Health and Medical Research Council/ ; 1132412//National Health and Medical Research Council/ ; 1118640//NHMRC Fellowship/ ; }, mesh = {*Aedes/drug effects/genetics/microbiology ; Animals ; Biological Control Agents ; Dengue/transmission ; Genotype ; Genotyping Techniques ; Indonesia ; Insect Proteins/genetics ; Insecticide Resistance ; Insecticides/pharmacology ; *Mosquito Control ; Mosquito Vectors/drug effects/genetics/microbiology ; Mutation ; Mutation Rate ; Pest Control, Biological ; Pyrethrins/pharmacology ; Voltage-Gated Sodium Channels/*genetics ; Wolbachia/pathogenicity ; }, abstract = {BACKGROUND: In the inner city of Yogyakarta, Indonesia, insecticide resistance is expected in the main dengue vector, Aedes aegypti, because of the intensive local application of pyrethroid insecticides. However, detailed information about the nature of resistance in this species is required to assist the release of Wolbachia mosquitoes in a dengue control program, so that we can ensure that insecticide resistance in the strain of Ae. aegypti being released matches that of the background population.

METHODS: High-resolution melt genotyping was used to screen for kdr mutations associated with pyrethroid resistance in the voltage-sensitive sodium channel (VSSC) gene in Ae. aegypti of some areas in the inner city of Yogyakarta.

RESULTS: The results show that the V1016G mutation predominated, with individuals homozygous for the 1016G allele at a frequency of 82.1% and the mutant allele G at a frequency of 92%. Two patterns of co-occurrence of mutations were detected in this study, homozygous individuals V1016G/S989P; and heterozygous individuals V1016G/F1534C/S989P. We found the simultaneous occurrence of kdr mutations V1016G and F1534C at all collection sites, but not within individual mosquitoes. Homozygous mutants at locus 1016 were homozygous wild-type at locus 1534 and vice versa, and heterozygous V1016G were also heterozygous for F1534C. The most common tri-locus genotype co-occurrences were homozygous mutant 1016GG and homozygous wild-type FF1534, combined with homozygous mutant 989PP (GG/FF/PP) at a frequency of 38.28%.

CONCLUSIONS: Given the relatively small differences in frequency of resistance alleles across the city area, locality variations in resistance should have minor implications for the success of Wolbachia mosquito trials being undertaken in the Yogyakarta area.}, } @article {pmid32824858, year = {2020}, author = {Bykov, R and Kerchev, I and Demenkova, M and Ryabinin, A and Ilinsky, Y}, title = {Sex-Specific Wolbachia Infection Patterns in Populations of Polygraphus proximus Blandford (Coleoptera; Curculionidae: Scolytinae).}, journal = {Insects}, volume = {11}, number = {8}, pages = {}, pmid = {32824858}, issn = {2075-4450}, support = {18-316-00099, 18-34-20060, 19-04-00983//Russian Foundation for Basic Research/ ; 0324-2019-0041//State Budgeted Project/ ; }, abstract = {Wolbachia symbionts are maternally inherited bacteria that are widely distributed among Arthropoda hosts. Wolbachia influence their host biology in diverse ways. They may induce reproductive abnormalities, protect hosts against pathogens and parasites, or benefit hosts through metabolic provisioning. The progeny of an infected female are ordinarily infected with Wolbachia; however, Wolbachia have no future in male host progeny because they cannot transmit the symbiont to the next generation. Here, we analyze native and invasive populations of the four-eyed fir bark beetle (Polygraphus proximus) for Wolbachia prevalence and symbiont genetic diversity. This species is a dangerous pest of Siberian fir (Abies sibirica) forests. The native range of P. proximus includes the territories of the Russian Far East, Japan, Korea, and Northeast China, whereas its invasive range includes West Siberia, with further expansion westward. Surprisingly, we revealed a difference in the patterns of Wolbachia prevalence for males and females. Infection rate and Wolbachia titers were higher in females than in males. ST-533, the only haplotype of Wolbachia supergroup B, was associated with a minimum of three out of the five described mitochondrial haplotypes.}, } @article {pmid32823761, year = {2020}, author = {Dally, M and Lalzar, M and Belausov, E and Gottlieb, Y and Coll, M and Zchori-Fein, E}, title = {Cellular Localization of Two Rickettsia Symbionts in the Digestive System and within the Ovaries of the Mirid Bug, Macrolophous pygmaeus.}, journal = {Insects}, volume = {11}, number = {8}, pages = {}, pmid = {32823761}, issn = {2075-4450}, abstract = {Bacterial symbionts in arthropods are common, vary in their effects, and can dramatically influence the outcome of biological control efforts. Macrolophus pygmaeus (Heteroptera: Miridae), a key component of biological control programs, is mainly predaceous but may also display phytophagy. M. pygmaeus hosts symbiotic Wolbachia, which induce cytoplasmic incompatibility, and two Rickettsia species, R. bellii and R. limoniae, which are found in all individuals tested. To test possible involvement of the two Rickettsia species in the feeding habits of M. pygmaeus, we first showed that the microbiome of the insect is dominated by these three symbionts, and later described the distribution pattern of the two Rickettsia species in its digestive system. Although both Rickettsia species were located in certain gut bacteriocyes, in caeca and in Malpighian tubules of both sexes, each species has a unique cellular occupancy pattern and specific distribution along digestive system compartments. Infrequently, both species were found in a cell. In females, both Rickettsia species were detected in the germarium, the apical end of the ovarioles within the ovaries, but not in oocytes. Although the cause for these Rickettsia distribution patterns is yet unknown, it is likely linked to host nutrition while feeding on prey or plants.}, } @article {pmid32823726, year = {2020}, author = {Ahmad, NA and Endersby-Harshman, NM and Mohd Mazni, NR and Mohd Zabari, NZA and Amran, SNS and Ridhuan Ghazali, MK and Abdul Karim, MA and Cheong, YL and Sinkins, SP and Ahmad, NW and Hoffmann, AA}, title = {Characterization of Sodium Channel Mutations in the Dengue Vector Mosquitoes Aedes aegypti and Aedes albopictus within the Context of Ongoing Wolbachia Releases in Kuala Lumpur, Malaysia.}, journal = {Insects}, volume = {11}, number = {8}, pages = {}, pmid = {32823726}, issn = {2075-4450}, support = {1037003, 1132412//NHMRC/ ; 108508/WT_/Wellcome Trust/United Kingdom ; Funding//Ministry of Health, Malaysia/ ; }, abstract = {Specific sodium channel gene mutations confer target site resistance to pyrethroid insecticides in mosquitoes and other insects. In Aedes mosquito species, multiple mutations that contribute to resistance vary in their importance around the world. Here, we characterize voltage sensitive sodium channel (Vssc) mutations in populations of Aedesaegypti from Kuala Lumpur, Malaysia, and look at their persistence in populations affected by ongoing Wolbachia releases (a dengue control measure). We also describe a Vssc mutation in Aedesalbopictus (F1534L) found for the first time in Malaysia. We show that there are three predominant Vssc haplotypes in Aedesaegypti in this region, which all persist with regular backcrossing, thereby maintaining the original genetic composition of the populations. We identify changes in genotype frequency in closed populations of Ae. aegypti maintained for multiple generations in laboratory culture, suggesting different fitness costs associated with the genotypes, some of which may be associated with the sex of the mosquito. Following population replacement of Ae. aegypti by Wolbachia in the target area, however, we find that the Vssc mutations have persisted at pre-release levels. Mosquitoes in two genotype classes demonstrate a type I pyrethroid resistance advantage over wildtype mosquitoes when exposed to 0.25% permethrin. This resistance advantage is even more pronounced with a type II pyrethroid, deltamethrin (0.03%). The results point to the importance of these mutations in pyrethroid resistance in mosquito populations and the need for regular backcrossing with male mosquitoes from the field to maintain similarity of genetic background and population integrity during Wolbachia releases.}, } @article {pmid32819378, year = {2020}, author = {Filipović, I and Hapuarachchi, HC and Tien, WP and Razak, MABA and Lee, C and Tan, CH and Devine, GJ and Rašić, G}, title = {Using spatial genetics to quantify mosquito dispersal for control programs.}, journal = {BMC biology}, volume = {18}, number = {1}, pages = {104}, pmid = {32819378}, issn = {1741-7007}, support = {Climate Resilience Studies Fund//Ministry of Finance/International ; Nectar Research Cloud//National Collaborative Research Infrastructure Strategy/International ; }, mesh = {Aedes/genetics/*physiology ; *Animal Distribution ; Animals ; Genetic Variation ; *Mosquito Control ; Mosquito Vectors/genetics/*physiology ; Singapore ; Spatial Analysis ; Time Factors ; }, abstract = {BACKGROUND: Hundreds of millions of people get a mosquito-borne disease every year and nearly one million die. Transmission of these infections is primarily tackled through the control of mosquito vectors. The accurate quantification of mosquito dispersal is critical for the design and optimization of vector control programs, yet the measurement of dispersal using traditional mark-release-recapture (MRR) methods is logistically challenging and often unrepresentative of an insect's true behavior. Using Aedes aegypti (a major arboviral vector) as a model and two study sites in Singapore, we show how mosquito dispersal can be characterized by the spatial analyses of genetic relatedness among individuals sampled over a short time span without interruption of their natural behaviors.

RESULTS: Using simple oviposition traps, we captured adult female Ae. aegypti across high-rise apartment blocks and genotyped them using genome-wide SNP markers. We developed a methodology that produces a dispersal kernel for distance which results from one generation of successful breeding (effective dispersal), using the distance separating full siblings and 2nd- and 3rd-degree relatives (close kin). The estimated dispersal distance kernel was exponential (Laplacian), with a mean dispersal distance (and dispersal kernel spread σ) of 45.2 m (95% CI 39.7-51.3 m), and 10% probability of a dispersal > 100 m (95% CI 92-117 m). Our genetically derived estimates matched the parametrized dispersal kernels from previous MRR experiments. If few close kin are captured, a conventional genetic isolation-by-distance analysis can be used, as it can produce σ estimates congruent with the close-kin method if effective population density is accurately estimated. Genetic patch size, estimated by spatial autocorrelation analysis, reflects the spatial extent of the dispersal kernel "tail" that influences, for example, the critical radii of release zones and the speed of Wolbachia spread in mosquito replacement programs.

CONCLUSIONS: We demonstrate that spatial genetics can provide a robust characterization of mosquito dispersal. With the decreasing cost of next-generation sequencing, the production of spatial genetic data is increasingly accessible. Given the challenges of conventional MRR methods, and the importance of quantified dispersal in operational vector control decisions, we recommend genetic-based dispersal characterization as the more desirable means of parameterization.}, } @article {pmid32813725, year = {2020}, author = {Shropshire, JD and Kalra, M and Bordenstein, SR}, title = {Evolution-guided mutagenesis of the cytoplasmic incompatibility proteins: Identifying CifA's complex functional repertoire and new essential regions in CifB.}, journal = {PLoS pathogens}, volume = {16}, number = {8}, pages = {e1008794}, pmid = {32813725}, issn = {1553-7374}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified/microbiology/physiology ; Bacterial Proteins/genetics/*metabolism ; *Biological Evolution ; Cytoplasm/*metabolism/microbiology ; Drosophila melanogaster/*microbiology/physiology ; Female ; Gram-Negative Bacterial Infections/*metabolism/microbiology/pathology ; Male ; *Mutation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are the world's most common, maternally-inherited, arthropod endosymbionts. Their worldwide distribution is due, in part, to a selfish drive system termed cytoplasmic incompatibility (CI) that confers a relative fitness advantage to females that transmit Wolbachia to their offspring. CI results in embryonic death when infected males mate with uninfected females but not infected females. Under the Two-by-One genetic model of CI, males expressing the two phage WO proteins CifA and CifB cause CI, and females expressing CifA rescue CI. While each protein is predicted to harbor three functional domains, there is no knowledge on how sites across these Cif domains, rather than in any one particular domain, contribute to CI and rescue. Here, we use evolution-guided, substitution mutagenesis of conserved amino acids across the Cif proteins, coupled with transgenic expression in uninfected Drosophila melanogaster, to determine the functional impacts of conserved residues evolving mostly under purifying selection. We report that amino acids in CifA's N-terminal unannotated region and annotated catalase-related domain are important for both complete CI and rescue, whereas C-terminal residues in CifA's putative domain of unknown function are solely important for CI. Moreover, conserved CifB amino acids in the predicted nucleases, peptidase, and unannotated regions are essential for CI. Taken together, these findings indicate that (i) all CifA amino acids determined to be crucial in rescue are correspondingly crucial in CI, (ii) an additional set of CifA amino acids are uniquely important in CI, and (iii) CifB amino acids across the protein, rather than in one particular domain, are all crucial for CI. We discuss how these findings advance an expanded view of Cif protein evolution and function, inform the mechanistic and biochemical bases of Cif-induced CI/rescue, and continue to substantiate the Two-by-One genetic model of CI.}, } @article {pmid32813707, year = {2020}, author = {Seabourn, P and Spafford, H and Yoneishi, N and Medeiros, M}, title = {The Aedes albopictus (Diptera: Culicidae) microbiome varies spatially and with Ascogregarine infection.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {8}, pages = {e0008615}, pmid = {32813707}, issn = {1935-2735}, support = {P20 GM125508/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*parasitology ; Animal Diseases ; Animals ; Apicomplexa ; Bacteria/*classification/genetics ; Host-Parasite Interactions ; Larva/microbiology/parasitology ; Microbiota/*physiology ; Protozoan Infections, Animal/*microbiology ; Symbiosis ; }, abstract = {The mosquito microbiome alters the physiological traits of medically important mosquitoes, which can scale to impact how mosquito populations sustain disease transmission. The mosquito microbiome varies significantly within individual mosquitoes and among populations, however the ecological and environmental factors that contribute to this variation are poorly understood. To further understand the factors that influence variation and diversity of the mosquito microbiome, we conducted a survey of the bacterial microbiome in the medically important mosquito, Aedes albopictus, on the high Pacific island of Maui, Hawai'i. We detected three bacterial Phyla and twelve bacterial families: Proteobacteria, Acitinobacteria, and Firmicutes; and Anaplasmataceae, Acetobacteraceae, Enterobacteriaceae, Burkholderiaceae, Xanthobacteraceae, Pseudomonadaceae, Streptomycetaceae, Staphylococcaceae, Xanthomonadaceae, Beijerinckiaceae, Rhizobiaceae, and Sphingomonadaceae. The Ae. albopictus bacterial microbiota varied among geographic locations, but temperature and rainfall were uncorrelated with this spatial variation. Infection status with an ampicomplexan pathosymbiont Ascogregarina taiwanensis was significantly associated with the composition of the Ae. albopictus bacteriome. The bacteriomes of mosquitoes with an A. taiwanensis infection were more likely to include several bacterial symbionts, including the most abundant lineage of Wolbachia sp. Other symbionts like Asaia sp. and several Enterobacteriaceae lineages were less prevalent in A. taiwanensis-infected mosquitoes. This highlights the possibility that inter- and intra-domain interactions may structure the Ae. albopictus microbiome.}, } @article {pmid32803130, year = {2020}, author = {Indriani, C and Tantowijoyo, W and Rancès, E and Andari, B and Prabowo, E and Yusdi, D and Ansari, MR and Wardana, DS and Supriyati, E and Nurhayati, I and Ernesia, I and Setyawan, S and Fitriana, I and Arguni, E and Amelia, Y and Ahmad, RA and Jewell, NP and Dufault, SM and Ryan, PA and Green, BR and McAdam, TF and O'Neill, SL and Tanamas, SK and Simmons, CP and Anders, KL and Utarini, A}, title = {Reduced dengue incidence following deployments of Wolbachia-infected Aedes aegypti in Yogyakarta, Indonesia: a quasi-experimental trial using controlled interrupted time series analysis.}, journal = {Gates open research}, volume = {4}, number = {}, pages = {50}, pmid = {32803130}, issn = {2572-4754}, support = {R56 AI134724/AI/NIAID NIH HHS/United States ; }, abstract = {Background: Ae. aegypti mosquitoes stably transfected with the intracellular bacterium Wolbachia pipientis (wMel strain) have been deployed for biocontrol of dengue and related arboviral diseases in multiple countries. Field releases in northern Australia have previously demonstrated near elimination of local dengue transmission from Wolbachia-treated communities, and pilot studies in Indonesia have demonstrated the feasibility and acceptability of the method. We conducted a quasi-experimental trial to evaluate the impact of scaled Wolbachia releases on dengue incidence in an endemic setting in Indonesia. Methods: In Yogyakarta City, Indonesia, following extensive community engagement, wMel Wolbachia-carrying mosquitoes were released every two weeks for 13-15 rounds over seven months in 2016-17, in a contiguous 5 km [2] area (population 65,000). A 3 km [2] area (population 34,000) on the opposite side of the city was selected a priori as an untreated control area. Passive surveillance data on notified hospitalised dengue patients was used to evaluate the epidemiological impact of Wolbachia deployments, using controlled interrupted time-series analysis. Results: Rapid and sustained introgression of wMel Wolbachia into local Ae. aegypti populations was achieved. Thirty-four dengue cases were notified from the intervention area and 53 from the control area (incidence 26 vs 79 per 100,000 person-years) during 24 months following Wolbachia deployment. This corresponded in the regression model to a 73% reduction in dengue incidence (95% confidence interval 49%,86%) associated with the Wolbachia intervention. Exploratory analysis including 6 months additional post-intervention observations showed a small strengthening of this effect (30 vs 115 per 100,000 person-years; 76% reduction in incidence, 95%CI 60%,86%). Conclusions: We demonstrate a significant reduction in dengue incidence following successful introgression of Wolbachia into local Ae. aegypti populations in an endemic setting in Indonesia. These findings are consistent with previous field trials in northern Australia, and support the effectiveness of this novel approach for dengue control.}, } @article {pmid32797213, year = {2021}, author = {Martinez, J and Klasson, L and Welch, JJ and Jiggins, FM}, title = {Life and Death of Selfish Genes: Comparative Genomics Reveals the Dynamic Evolution of Cytoplasmic Incompatibility.}, journal = {Molecular biology and evolution}, volume = {38}, number = {1}, pages = {2-15}, pmid = {32797213}, issn = {1537-1719}, support = {/WT_/Wellcome Trust/United Kingdom ; 281668/ERC_/European Research Council/International ; WT202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; WT094664MA/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila/microbiology/physiology ; *Evolution, Molecular ; Female ; *Genome, Bacterial ; Host-Pathogen Interactions/*genetics ; Male ; *Repetitive Sequences, Nucleic Acid ; Spermatozoa/physiology ; Wolbachia/*genetics ; }, abstract = {Cytoplasmic incompatibility is a selfish reproductive manipulation induced by the endosymbiont Wolbachia in arthropods. In males Wolbachia modifies sperm, leading to embryonic mortality in crosses with Wolbachia-free females. In females, Wolbachia rescues the cross and allows development to proceed normally. This provides a reproductive advantage to infected females, allowing the maternally transmitted symbiont to spread rapidly through host populations. We identified homologs of the genes underlying this phenotype, cifA and cifB, in 52 of 71 new and published Wolbachia genome sequences. They are strongly associated with cytoplasmic incompatibility. There are up to seven copies of the genes in each genome, and phylogenetic analysis shows that Wolbachia frequently acquires new copies due to pervasive horizontal transfer between strains. In many cases, the genes have subsequently acquired loss-of-function mutations to become pseudogenes. As predicted by theory, this tends to occur first in cifB, whose sole function is to modify sperm, and then in cifA, which is required to rescue the cross in females. Although cif genes recombine, recombination is largely restricted to closely related homologs. This is predicted under a model of coevolution between sperm modification and embryonic rescue, where recombination between distantly related pairs of genes would create a self-incompatible strain. Together, these patterns of gene gain, loss, and recombination support evolutionary models of cytoplasmic incompatibility.}, } @article {pmid32796890, year = {2020}, author = {Straub, TJ and Shaw, WR and Marcenac, P and Sawadogo, SP and Dabiré, RK and Diabaté, A and Catteruccia, F and Neafsey, DE}, title = {The Anopheles coluzzii microbiome and its interaction with the intracellular parasite Wolbachia.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {13847}, pmid = {32796890}, issn = {2045-2322}, support = {U19 AI110818/AI/NIAID NIH HHS/United States ; U19AI110818//National Institute of Allergy and Infectious Diseases/International ; R21AI117313//National Institute of Allergy and Infectious Diseases/International ; }, mesh = {Animals ; Anopheles/*microbiology/*physiology ; Burkina Faso ; Disease Vectors ; Female ; Host Microbial Interactions/*physiology ; *Insect Vectors ; Malaria/transmission ; Mosquito Control/methods ; Oviposition ; Vector Borne Diseases/transmission ; *Wolbachia ; }, abstract = {Wolbachia, an endosymbiotic alpha-proteobacterium commonly found in insects, can inhibit the transmission of human pathogens by mosquitoes. Biocontrol programs are underway using Aedes aegypti mosquitoes trans-infected with a non-natural Wolbachia strain to reduce dengue virus transmission. Less is known about the impact of Wolbachia on the biology and vectorial capacity of Anopheles mosquitoes, the vectors of malaria parasites. A naturally occurring strain of Wolbachia, wAnga, infects populations of the major malaria vectors Anopheles gambiae and Anopheles coluzzii in Burkina Faso. Previous studies found wAnga infection was negatively correlated with Plasmodium infection in the mosquito and wAnga influenced mosquito egg-laying behavior. Here, we investigate wAnga in natural populations of An. coluzzii and its interactions with other resident microbiota using targeted 16S sequencing. Though we find no major differences in microbiota composition associated with wAnga infection, we do find several taxa that correlate with the presence or absence of wAnga in female mosquitoes following oviposition, with the caveat that we could not rule out batch effects due to the unanticipated impact of wAnga on oviposition timing. These data suggest wAnga may influence or interact with the Anopheles microbiota, which may contribute to the impact of wAnga on Anopheles biology and vectorial capacity.}, } @article {pmid32792343, year = {2020}, author = {Ferguson, KB and Kursch-Metz, T and Verhulst, EC and Pannebakker, BA}, title = {Hybrid Genome Assembly and Evidence-Based Annotation of the Egg Parasitoid and Biological Control Agent Trichogramma brassicae.}, journal = {G3 (Bethesda, Md.)}, volume = {10}, number = {10}, pages = {3533-3540}, pmid = {32792343}, issn = {2160-1836}, mesh = {Animals ; Biological Control Agents ; Germany ; *Hymenoptera/genetics ; *Moths ; Random Amplified Polymorphic DNA Technique ; *Wasps/genetics ; }, abstract = {Trichogramma brassicae (Bezdenko) are egg parasitoids that are used throughout the world as biological control agents and in laboratories as model species. Despite this ubiquity, few genetic resources exist beyond COI, ITS2, and RAPD markers. Aided by a Wolbachia infection, a wild-caught strain from Germany was reared for low heterozygosity and sequenced in a hybrid de novo strategy, after which several assembling strategies were evaluated. The best assembly, derived from a DBG2OLC-based pipeline, yielded a genome of 235 Mbp made up of 1,572 contigs with an N50 of 556,663 bp. Following a rigorous ab initio-, homology-, and evidence-based annotation, 16,905 genes were annotated and functionally described. As an example of the utility of the genome, a simple ortholog cluster analysis was performed with sister species T. pretiosum, revealing over 6000 shared clusters and under 400 clusters unique to each species. The genome and transcriptome presented here provides an essential resource for comparative genomics of the commercially relevant genus Trichogramma, but also for research into molecular evolution, ecology, and breeding of T. brassicae.}, } @article {pmid32788982, year = {2020}, author = {N Miyata, M and Nomura, M and Kageyama, D}, title = {Wolbachia have made it twice: Hybrid introgression between two sister species of Eurema butterflies.}, journal = {Ecology and evolution}, volume = {10}, number = {15}, pages = {8323-8330}, pmid = {32788982}, issn = {2045-7758}, abstract = {Wolbachia, cytoplasmically inherited endosymbionts of arthropods, are known to hijack their host reproduction in various ways to increase their own vertical transmission. This may lead to the selective sweep of associated mitochondria, which can have a large impact on the evolution of mitochondrial lineages. In Japan, two different Wolbacahia strains (wCI and wFem) are found in two sister species of pierid butterflies, Eurema mandarina and Eurema hecabe. In both species, females infected with wCI (C females) produce offspring with a nearly 1:1 sex ratio, while females infected with both wCI and wFem (CF females) produce all-female offspring. Previous studies have suggested the historical occurrence of hybrid introgression in C individuals between the two species. Furthermore, hybrid introgression in CF individuals is suggested by the distinct mitochondrial lineages between C females and CF females of E. mandarina. In this study, we performed phylogenetic analyses based on nuclear DNA and mitochondrial DNA markers of E. hecabe with previously published data on E. mandarina. We found that the nuclear DNA of this species significantly diverged from that of E. mandarina. By contrast, mitochondrial DNA haplotypes comprised two clades, mostly reflecting Wolbachia infection status rather than the individual species. Collectively, our results support the previously suggested occurrence of two independent historical events wherein the cytoplasms of CF females and C females moved between E. hecabe and E. mandarina through hybrid introgression.}, } @article {pmid32787974, year = {2020}, author = {Wong, ML and Liew, JWK and Wong, WK and Pramasivan, S and Mohamed Hassan, N and Wan Sulaiman, WY and Jeyaprakasam, NK and Leong, CS and Low, VL and Vythilingam, I}, title = {Natural Wolbachia infection in field-collected Anopheles and other mosquito species from Malaysia.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {414}, pmid = {32787974}, issn = {1756-3305}, support = {LR002C-2018//Ministry of Education Malaysia/ ; }, mesh = {Aedes/microbiology ; Animals ; Anopheles/microbiology ; Bacterial Outer Membrane Proteins/genetics ; Culex/microbiology ; Culicidae/*microbiology ; Genes, Bacterial ; Insect Control ; Malaysia/epidemiology ; Malvaceae/microbiology ; Mosquito Vectors/microbiology ; Pathology, Molecular ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Vector Borne Diseases/prevention & control ; *Wolbachia/genetics/isolation & purification ; }, abstract = {BACKGROUND: The endosymbiont bacterium Wolbachia is maternally inherited and naturally infects some filarial nematodes and a diverse range of arthropods, including mosquito vectors responsible for disease transmission in humans. Previously, it has been found infecting most mosquito species but absent in Anopheles and Aedes aegypti. However, recently these two mosquito species were found to be naturally infected with Wolbachia. We report here the extent of Wolbachia infections in field-collected mosquitoes from Malaysia based on PCR amplification of the Wolbachia wsp and 16S rRNA genes.

METHODS: The prevalence of Wolbachia in Culicinae mosquitoes was assessed via PCR with wsp primers. For some of the mosquitoes, in which the wsp primers failed to amplify a product, Wolbachia screening was performed using nested PCR targeting the 16S rRNA gene. Wolbachia sequences were aligned using Geneious 9.1.6 software, analyzed with BLAST, and the most similar sequences were downloaded. Phylogenetic analyses were carried out with MEGA 7.0 software. Graphs were drawn with GraphPad Prism 8.0 software.

RESULTS: A total of 217 adult mosquitoes representing 26 mosquito species were screened. Of these, infections with Wolbachia were detected in 4 and 15 mosquito species using wsp and 16S rRNA primers, respectively. To our knowledge, this is the first time Wolbachia was detected using 16S rRNA gene amplification, in some Anopheles species (some infected with Plasmodium), Culex sinensis, Culex vishnui, Culex pseudovishnui, Mansonia bonneae and Mansonia annulifera. Phylogenetic analysis based on wsp revealed Wolbachia from most of the mosquitoes belonged to Wolbachia Supergroup B. Based on 16S rRNA phylogenetic analysis, the Wolbachia strain from Anopheles mosquitoes were more closely related to Wolbachia infecting Anopheles from Africa than from Myanmar.

CONCLUSIONS: Wolbachia was found infecting Anopheles and other important disease vectors such as Mansonia. Since Wolbachia can affect its host by reducing the life span and provide resistance to pathogen infection, several studies have suggested it as a potential innovative tool for vector/vector-borne disease control. Therefore, it is important to carry out further studies on natural Wolbachia infection in vector mosquitoes' populations as well as their long-term effects in new hosts and pathogen suppression.}, } @article {pmid32784948, year = {2020}, author = {Sucupira, PHF and Ferreira, ÁGA and Leite, THJF and de Mendonça, SF and Ferreira, FV and Rezende, FO and Marques, JT and Moreira, LA}, title = {The RNAi Pathway Is Important to Control Mayaro Virus Infection in Aedes aegypti but not for Wolbachia-Mediated Protection.}, journal = {Viruses}, volume = {12}, number = {8}, pages = {}, pmid = {32784948}, issn = {1999-4915}, mesh = {Aedes/immunology/*microbiology/*virology ; Alphavirus/*genetics ; Alphavirus Infections/immunology/virology ; Animals ; Female ; Humans ; Immunity, Innate ; Mosquito Vectors/immunology/microbiology/virology ; *RNA Interference ; RNA, Small Interfering/*genetics/*metabolism ; Wolbachia/immunology/*physiology ; }, abstract = {Mayaro virus (MAYV), a sylvatic arbovirus belonging to the Togaviridae family and Alphavirus genus, is responsible for an increasing number of outbreaks in several countries of Central and South America. Despite Haemagogus janthinomys being identified as the main vector of MAYV, laboratory studies have already demonstrated the competence of Aedes aegypti to transmit MAYV. It has also been demonstrated that the WolbachiawMel strain is able to impair the replication and transmission of MAYV in Ae. aegypti. In Ae. aegypti, the small interfering RNA (siRNA) pathway is an important antiviral mechanism; however, it remains unclear whether siRNA pathway acts against MAYV infection in Ae. aegypti. The main objective of this study was to determine the contribution of the siRNA pathway in the control of MAYV infection. Thus, we silenced the expression of AGO2, an essential component of the siRNA pathway, by injecting dsRNA-targeting AGO2 (dsAGO2). Our results showed that AGO2 is required to control MAYV replication upon oral infection in Wolbachia-free Ae. aegypti. On the other hand, we found that Wolbachia-induced resistance to MAYV in Ae. aegypti is independent of the siRNA pathway. Our study brought new information regarding the mechanism of viral protection, as well as on Wolbachia mediated interference.}, } @article {pmid32783512, year = {2020}, author = {Shi, Y and Yu, J}, title = {Wolbachia infection enhancing and decaying domains in mosquito population based on discrete models.}, journal = {Journal of biological dynamics}, volume = {14}, number = {1}, pages = {679-695}, doi = {10.1080/17513758.2020.1805035}, pmid = {32783512}, issn = {1751-3766}, mesh = {Animals ; Bacterial Infections/epidemiology/*microbiology ; Culicidae/*microbiology ; Female ; *Models, Biological ; Numerical Analysis, Computer-Assisted ; Wolbachia/*physiology ; }, abstract = {In this article, we formulate and study a discrete equation model depicting the pattern of Wolbachia infection in a mosquito population. A domain in [Formula: see text] is called a Wolbachia infection enhancing (or decaying) domain if in which the Wolbachia infection frequency of the next generation is always bigger (or smaller) than that of the current generation. We first give a complete analysis of the equivalent Wolbachia infection frequency curves. And then we clearly characterize the Wolbachia infection enhancing domain and decaying domain for all of the parameters, respectively. Finally, some numerical examples are also provided to illustrate our theoretical results.}, } @article {pmid32779567, year = {2020}, author = {Curran, DM and Grote, A and Nursimulu, N and Geber, A and Voronin, D and Jones, DR and Ghedin, E and Parkinson, J}, title = {Modeling the metabolic interplay between a parasitic worm and its bacterial endosymbiont allows the identification of novel drug targets.}, journal = {eLife}, volume = {9}, number = {}, pages = {}, pmid = {32779567}, issn = {2050-084X}, support = {R21 AI126466/AI/NIAID NIH HHS/United States ; RGPIN-2014-06664//Natural Sciences and Engineering Research Council of Canada/International ; T32 AI007180/AI/NIAID NIH HHS/United States ; F31 AI131527/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*drug effects/microbiology ; *Drug Evaluation, Preclinical ; Filariasis/*drug therapy ; Filaricides/*pharmacology ; Metabolic Networks and Pathways/drug effects ; Models, Biological ; *Symbiosis/drug effects ; Wolbachia/*drug effects ; }, abstract = {The filarial nematode Brugia malayi represents a leading cause of disability in the developing world, causing lymphatic filariasis in nearly 40 million people. Currently available drugs are not well-suited to mass drug administration efforts, so new treatments are urgently required. One potential vulnerability is the endosymbiotic bacteria Wolbachia-present in many filariae-which is vital to the worm. Genome scale metabolic networks have been used to study prokaryotes and protists and have proven valuable in identifying therapeutic targets, but have only been applied to multicellular eukaryotic organisms more recently. Here, we present iDC625, the first compartmentalized metabolic model of a parasitic worm. We used this model to show how metabolic pathway usage allows the worm to adapt to different environments, and predict a set of 102 reactions essential to the survival of B. malayi. We validated three of those reactions with drug tests and demonstrated novel antifilarial properties for all three compounds.}, } @article {pmid32774328, year = {2020}, author = {Duan, R and Xu, H and Gao, S and Gao, Z and Wang, N}, title = {Effects of Different Hosts on Bacterial Communities of Parasitic Wasp Nasonia vitripennis.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {1435}, pmid = {32774328}, issn = {1664-302X}, abstract = {Parasitism is a special interspecific relationship in insects. Unlike most other ectoparasites, Nasonia vitripennis spend most of its life cycle (egg, larvae, pupae, and early adult stage) inside the pupae of flies, which is covered with hard puparium. Microbes play important roles in host development and help insect hosts to adapt to various environments. How the microbes of parasitic wasp respond to different fly hosts living in such close relationships motivated this investigation. In this study, we used N. vitripennis and three different fly pupa hosts (Lucilia sericata, Sarcophaga marshalli, and Musca domestica) to address this question, as well as to illustrate the potential transfer of bacteria through the trophic food chains. We found that N. vitripennis from different fly pupa hosts showed distinct microbiota, which means that the different fly hosts could affect the bacterial communities of their parasitic wasps. Some bacteria showed potential horizontal transfer through the trophic food chains, from the food through the fly to the parasitic wasp. We also found that the heritable endosymbiont Wolbachia could transferred from the fly host to the parasite and correlated with the bacterial communities of the corresponding parasitic wasps. Our findings provide new insight to the microbial interactions between parasite and host.}, } @article {pmid32770272, year = {2021}, author = {Thongprem, P and Davison, HR and Thompson, DJ and Lorenzo-Carballa, MO and Hurst, GDD}, title = {Incidence and Diversity of Torix Rickettsia-Odonata Symbioses.}, journal = {Microbial ecology}, volume = {81}, number = {1}, pages = {203-212}, pmid = {32770272}, issn = {1432-184X}, support = {CGL2008-02799//Ministerio de Ciencia e Innovación/ ; }, mesh = {Animals ; DNA Barcoding, Taxonomic ; Female ; Infectious Disease Transmission, Vertical ; Odonata/*microbiology ; Ovary/microbiology ; Rickettsia/classification/genetics/*physiology ; Rickettsia Infections/*transmission ; Symbiosis/*physiology ; }, abstract = {Heritable microbes are an important component of invertebrate biology, acting both as beneficial symbionts and reproductive parasites. Whilst most previous research has focussed on the 'Wolbachia pandemic', recent work has emphasised the importance of other microbial symbionts. In this study, we present a survey of odonates (dragonflies and damselflies) for torix group Rickettsia, following previous research indicating that this clade can be common in other aquatic insect groups. PCR assays were used to screen a broad range of odonates from two continents and revealed 8 of 76 species tested were infected with Rickettsia. We then conducted further deeper screening of UK representatives of the Coenagrionidae damselfly family, revealing 6 of 8 UK coenagrionid species to be positive for torix Rickettsia. Analysis of Rickettsia gene sequences supported multiple establishments of symbiosis in the group. Some strains were shared between UK coenagrionid species that shared mtDNA barcodes, indicating a likely route for mitochondrial introgression between sister species. There was also evidence of coinfecting Rickettsia strains in two species. FISH analysis indicated Rickettsia were observed in the ovarioles, consistent with heritable symbiosis. We conclude that torix Rickettsia represent an important associate of odonates, being found in a broad range of species from both Europe and South America. There is evidence that coinfection can occur, vertical transmission is likely, and that symbiont movement following hybridisation may underpin the lack of 'barcoding gap' between well-established species pairs in the genus. Future work should establish the biological significance of the symbioses observed.}, } @article {pmid32765466, year = {2020}, author = {Liang, X and Liu, J and Bian, G and Xi, Z}, title = {Wolbachia Inter-Strain Competition and Inhibition of Expression of Cytoplasmic Incompatibility in Mosquito.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {1638}, pmid = {32765466}, issn = {1664-302X}, abstract = {Successful field trials have been reported as part of the effort to develop the maternally transmitted endosymbiontic bacteria Wolbachia as an intervention agent for controlling mosquito vectors and their transmitted diseases. In order to further improve this novel intervention, artificially transinfected mosquitoes must be optimized to display maximum pathogen blocking, the desired cytoplasmic incompatibility (CI) pattern, and the lowest possible fitness cost. Achieving such optimization, however, requires a better understanding of the interactions between the host and various Wolbabachia strains and their combinations. Here, we transferred the Wolbachia wMel strain by embryonic microinjection into Aedes albopictus, resulting in the successful establishment of a transinfected line, HM (wAlbAwAlbBwMel), with a triple-strain infection comprising wMel, wAlbA, and wAlbB. Surprisingly, no CI was induced when the triply infected males were crossed with the wild-type GUA females or with another triply infected HC females carrying wPip, wAlbA, and wAlbB, but specific removal of wAlbA from the HM (wAlbAwAlbBwMel) line resulted in the expression of CI after crosses with lines infected by either one, two, or three strains of Wolbachia. The transinfected line showed perfect maternal transmission of the triple infection, with fluctuating egg hatch rates that improved to normal levels after repeated outcrosses with GUA line. Strain-specific qPCR assays showed that wMel and wAlbB were present at the highest densities in the ovaries and midguts, respectively, of the HM (wAlbAwAlbBwMel) mosquitoes. These finding suggest that introducing a novel strain of Wolbachia into a Wolbachia-infected host may result in complicated interactions between Wolbachia and the host and between the various Wolbachia strains, with competition likely to occur between strains in the same supergroup.}, } @article {pmid32759653, year = {2020}, author = {Dew, RM and McFrederick, QS and Rehan, SM}, title = {Diverse Diets with Consistent Core Microbiome in Wild Bee Pollen Provisions.}, journal = {Insects}, volume = {11}, number = {8}, pages = {}, pmid = {32759653}, issn = {2075-4450}, support = {1004515//U.S. Department of Agriculture/ ; CA-R-ENT-5109-H//U.S. Department of Agriculture/ ; 549038//Foundation for Food and Agriculture Research/ ; }, abstract = {Bees collect pollen from flowers for their offspring, and by doing so contribute critical pollination services for our crops and ecosystems. Unlike many managed bee species, wild bees are thought to obtain much of their microbiome from the environment. However, we know surprisingly little about what plant species bees visit and the microbes associated with the collected pollen. Here, we addressed the hypothesis that the pollen and microbial components of bee diets would change across the range of the bee, by amplicon sequencing pollen provisions of a widespread small carpenter bee, Ceratina calcarata, across three populations. Ceratina calcarata was found to use a diversity of floral resources across its range, but the bacterial genera associated with pollen provisions were very consistent. Acinetobacter, Erwinia, Lactobacillus, Sodalis, Sphingomonas and Wolbachia were among the top ten bacterial genera across all sites. Ceratina calcarata uses both raspberry (Rubus) and sumac (Rhus) stems as nesting substrates, however nests within these plants showed no preference for host plant pollen. Significant correlations in plant and bacterial co-occurrence differed between sites, indicating that many of the most common bacterial genera have either regional or transitory floral associations. This range-wide study suggests microbes present in brood provisions are conserved within a bee species, rather than mediated by climate or pollen composition. Moving forward, this has important implications for how these core bacteria affect larval health and whether these functions vary across space and diet. These data increase our understanding of how pollinators interact with and adjust to their changing environment.}, } @article {pmid32752052, year = {2020}, author = {Laidoudi, Y and Medkour, H and Levasseur, A and Davoust, B and Mediannikov, O}, title = {New Molecular Data on Filaria and its Wolbachia from Red Howler Monkeys (Alouatta macconnelli) in French Guiana-A Preliminary Study.}, journal = {Pathogens (Basel, Switzerland)}, volume = {9}, number = {8}, pages = {}, pmid = {32752052}, issn = {2076-0817}, abstract = {Previous studies have reported filarial parasites of the genus Dipetalonema and Mansonella from French Guiana monkeys, based on morphological taxonomy. In this study, we screened blood samples from nine howler monkeys (Alouatta macconnelli) for the presence of filaria and Wolbachia DNA. The infection rates were 88.9% for filaria and 55.6% for wolbachiae. The molecular characterization, based on the 18S gene of filariids, revealed that A. macconnelli are infected with at least three species (Mansonella sp., Brugia sp. and an unidentified Onchocercidae species.). Since the 18S and cox1 generic primers are not very effective at resolving co-infections, we developed ITS genus-specific PCRs for Mansonella and Brugia genus. The results revealed coinfections in 75% of positives. The presence of Mansonella sp. and Brugia sp. was also confirmed by the 16S phylogenetic analysis of their associated Wolbachia. Mansonella sp., which close to the species from the subgenus Tetrapetalonema encountered in New World Monkeys, while Brugia sp. was identical to the strain circulating in French Guiana dogs. We propose a novel ITS1Brugia genus-specific qPCR. We applied it to screen for Brugia infection in howler monkeys and 66.7% were found to be positive. Our finding highlights the need for further studies to clarify the species diversity of neotropics monkeys by combining molecular and morphological features. The novel Brugia genus-specific qPCR assays could be an effective tool for the surveillance and characterization of this potential zoonosis.}, } @article {pmid32748704, year = {2020}, author = {Huang, M and Hu, L}, title = {Modeling the suppression dynamics of Aedes mosquitoes with mating inhomogeneity.}, journal = {Journal of biological dynamics}, volume = {14}, number = {1}, pages = {656-678}, doi = {10.1080/17513758.2020.1799083}, pmid = {32748704}, issn = {1751-3766}, mesh = {Aedes/*physiology ; Animals ; Dengue/epidemiology/parasitology ; Incidence ; *Models, Biological ; Population Dynamics ; Seasons ; Sexual Behavior, Animal/*physiology ; Temperature ; }, abstract = {A novel strategy for controlling mosquito-borne diseases, such as dengue, malaria and Zika, involves releases of Wolbachia-infected mosquitoes as Wolbachia cause early embryo death when an infected male mates with an uninfected female. In this work, we introduce a delay differential equation model with mating inhomogeneity to discuss mosquito population suppression based on Wolbachia. Our analyses show that the wild mosquitoes could be eliminated if either the adult mortality rate exceeds the threshold [Formula: see text] or the release amount exceeds the threshold [Formula: see text] uniformly. We also present the nonlinear dependence of [Formula: see text] and [Formula: see text] on the parameters, respectively, as well as the effect of pesticide spraying on wild mosquitoes. Our simulations suggest that the releasing should be started at least 5 weeks before the peak dengue season, taking into account both the release amount and the suppression speed.}, } @article {pmid32748037, year = {2020}, author = {Ashour, DS and Othman, AA}, title = {Parasite-bacteria interrelationship.}, journal = {Parasitology research}, volume = {119}, number = {10}, pages = {3145-3164}, pmid = {32748037}, issn = {1432-1955}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Arthropods/microbiology ; Bacterial Infections/*complications ; Filarioidea/*microbiology ; Humans ; Parasites/microbiology ; Parasitic Diseases/*complications ; Probiotics/therapeutic use ; Schistosoma haematobium/*microbiology ; Symbiosis ; Urinary Bladder Neoplasms/microbiology/parasitology/pathology ; Wolbachia/*growth & development ; }, abstract = {Parasites and bacteria have co-evolved with humankind, and they interact all the time in a myriad of ways. For example, some bacterial infections result from parasite-dwelling bacteria as in the case of Salmonella infection during schistosomiasis. Other bacteria synergize with parasites in the evolution of human disease as in the case of the interplay between Wolbachia endosymbiont bacteria and filarial nematodes as well as the interaction between Gram-negative bacteria and Schistosoma haematobium in the pathogenesis of urinary bladder cancer. Moreover, secondary bacterial infections may complicate several parasitic diseases such as visceral leishmaniasis and malaria, due to immunosuppression of the host during parasitic infections. Also, bacteria may colonize the parasitic lesions; for example, hydatid cysts and skin lesions of ectoparasites. Remarkably, some parasitic helminths and arthropods exhibit antibacterial activity usually by the release of specific antimicrobial products. Lastly, some parasite-bacteria interactions are induced as when using probiotic bacteria to modulate the outcome of a variety of parasitic infections. In sum, parasite-bacteria interactions involve intricate processes that never cease to intrigue the researchers. However, understanding and exploiting these interactions could have prophylactic and curative potential for infections by both types of pathogens.}, } @article {pmid32741378, year = {2020}, author = {Wang, Z and Li, H and Zhou, X and Tang, M and Sun, L and Zhan, S and Xiao, Q}, title = {Comparative characterization of microbiota between the sibling species of tea geometrid moth Ectropis obliqua Prout and E. grisescens Warren.}, journal = {Bulletin of entomological research}, volume = {110}, number = {6}, pages = {684-693}, doi = {10.1017/S0007485320000164}, pmid = {32741378}, issn = {1475-2670}, mesh = {Animals ; Bacteria/classification/isolation & purification ; Female ; Male ; *Microbiota ; Moths/*microbiology ; Reproductive Isolation ; Symbiosis ; }, abstract = {For a wide range of insect species, the microbiota has potential roles in determining host developmental programme, immunity and reproductive biology. The tea geometrid moths Ectropis obliqua and E. grisescens are two closely related species that mainly feed on tea leaves. Although they can mate, infertile hybrids are produced. Therefore, these species provide a pair of model species for studying the molecular mechanisms of microbiotal involvement in host reproductive biology. In this study, we first identified and compared the compositions of microbiota between these sibling species, revealing higher microbiotal diversity for E. grisescens. The microbiota of E. obliqua mainly comprised the phyla Firmicutes, Proteobacteria and Cyanobacteria, whereas that of E. grisescens was dominated by Proteobacteria, Actinobacteria and Firmicutes. At the genus level, the dominant microbiota of E. grisescens included Wolbachia, Enterobacter and Pseudomonas and that of E. obliqua included Melissococcus, Staphylococcus and Enterobacter. Furthermore, we verified the rate of Wolbachia to infect 80 samples from eight different geographical populations, and the results supported that only E. grisescens harboured Wolbachia. Taken together, our findings indicate significantly different microbiotal compositions for E. obliqua and E. grisescens, with Wolbachia possibly being a curial factor influencing the reproductive isolation of these species. This study provides new insight into the mechanisms by which endosymbiotic bacteria, particularly Wolbachia, interact with sibling species.}, } @article {pmid32741005, year = {2020}, author = {Zheng, XL}, title = {Unveiling mosquito cryptic species and their reproductive isolation.}, journal = {Insect molecular biology}, volume = {29}, number = {6}, pages = {499-510}, pmid = {32741005}, issn = {1365-2583}, mesh = {Animals ; Culicidae/microbiology/*physiology ; *Genetic Speciation ; Mosquito Vectors/microbiology/*physiology ; *Reproductive Isolation ; *Sympatry ; Wolbachia/physiology ; }, abstract = {Mosquitoes are major vectors of many infectious pathogens or parasites. Understanding cryptic species and the speciation of disease vectors has important implications for vector management, evolution and host-pathogen and/or host-parasite interactions. Currently, mosquito cryptic species have been reported in many studies, most of which focus on the reproductive isolation of cryptic species and mainly on Anopheles gambiae sensu lato complex. Emerging species within the primary malaria vector Anopheles gambiae show different ecological preferences and significant prezygotic reproductive isolation, while Aedes mariae and Aedes zammitii show postmating reproductive isolation. However, data reporting the reproductive isolation in Culex and Aedes albopictus mosquito cryptic species is absent. The lack of systematic studies leaves many questions open, such as whether cryptic species are more common in particular habitats, latitudes or taxonomic groups; what mosquito cryptic species evolutionary processes bring about reproductive isolation in the absence of morphological differentiation? How does Wolbachia infection affect in mosquitoes' reproductive isolation? In this review, we provide a summary of recent advances in the discovery and identification of sibling or cryptic species within mosquito genera.}, } @article {pmid32737539, year = {2021}, author = {Takamatsu, T and Arai, H and Abe, N and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Coexistence of Two Male-Killers and Their Impact on the Development of Oriental Tea Tortrix Homona magnanima.}, journal = {Microbial ecology}, volume = {81}, number = {1}, pages = {193-202}, pmid = {32737539}, issn = {1432-184X}, support = {24580076//Japan Society for the Promotion of Science/ ; 19J13123//Japan Society for the Promotion of Science/ ; }, mesh = {Animals ; Female ; Male ; Moths/*microbiology ; RNA Virus Infections/*mortality ; RNA Viruses/*pathogenicity ; Reproduction/*physiology ; Spiroplasma/*physiology ; Wolbachia/metabolism ; }, abstract = {Male-killing, the death of male offspring induced by maternally transmitted microbes, is classified as early, or late, male-killing. The primary advantage afforded by early male-killing, which typically occurs during embryogenesis, is the reallocation of resources to females, that would have otherwise been consumed by males. Meanwhile, the key advantage of late male-killing, which typically occurs during late larval development, is the maximized potential for horizontal transmission. To date, no studies have reported on the associated developmental and physiological effects of host coinfection with early and late male-killers, which may have a significant impact on the population dynamics of the male-killers. Here we used a lepidopteran tea pest Homona magnanima as a model, which is a unique system wherein an early male-killer (a Spiroplasma bacterium) and a late male-killer (an RNA virus) can coexist in nature. An artificially established matriline, coinfected with both Spiroplasma and RNA virus, exhibited embryonic death (early male-killing) as seen in the host line singly infected with Spiroplasma. Moreover, the coinfected line also exhibited developmental retardation and low pupal weight similar to the host line singly infected with the RNA virus. A series of field surveys revealed that Spiroplasma-RNA virus coinfection occurs in nature at a low frequency. Hence, although the two male-killers are capable of coexisting within the H. magnanima population independently, high associated fitness cost appears to limit the prevalence of male-killer coinfection in the field host population.}, } @article {pmid32733407, year = {2020}, author = {Ford, SA and Albert, I and Allen, SL and Chenoweth, SF and Jones, M and Koh, C and Sebastian, A and Sigle, LT and McGraw, EA}, title = {Artificial Selection Finds New Hypotheses for the Mechanism of Wolbachia-Mediated Dengue Blocking in Mosquitoes.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {1456}, pmid = {32733407}, issn = {1664-302X}, support = {R01 AI143758/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia is an intracellular bacterium that blocks virus replication in insects and has been introduced into the mosquito, Aedes aegypti for the biocontrol of arboviruses including dengue, Zika, and chikungunya. Despite ongoing research, the mechanism of Wolbachia-mediated virus blocking remains unclear. We recently used experimental evolution to reveal that Wolbachia-mediated dengue blocking could be selected upon in the A. aegypti host and showed evidence that strong levels of blocking could be maintained by natural selection. In this study, we investigate the genetic variation associated with blocking and use these analyses to generate testable hypotheses surrounding the mechanism of Wolbachia-mediated dengue blocking. From our results, we hypothesize that Wolbachia may block virus replication by increasing the regeneration rate of mosquito cells via the Notch signaling pathway. We also propose that Wolbachia modulates the host's transcriptional pausing pathway either to prime the host's anti-viral response or to directly inhibit viral replication.}, } @article {pmid32733140, year = {2020}, author = {Wöger, R and Wöger, R and Nuss, M}, title = {Spatial and temporal sex ratio bias and Wolbachia-infection in New Zealand Crambidae (Lepidoptera: Pyraloidea).}, journal = {Biodiversity data journal}, volume = {8}, number = {}, pages = {e52621}, pmid = {32733140}, issn = {1314-2828}, abstract = {The New Zealand fauna of snout moths (Pyraloidea) predominantly consists of endemic species. During 2017 and 2018, 56 species of Pyraloidea in 1,749 individuals were collected at 14 localities. All species were screened for Wolbachia-infection, with specimens of eight species (14%) being positive, of which six species belong to Scopariinae. This is the first record of Wolbachia-infection amongst New Zealand Lepidoptera. The most common pyraloid species, Eudonia submarginalis and Orocrambus flexuosellus, were analysed for a larger set of individuals looking for sex ratio and Wolbachia-infection. There is a sex ratio bias towards females in both species, but it varies in space and time. Wolbachia is found in all populations of E. submarginalis with 10-80% of the tested individuals being positive, depending on locality. No Wolbachia-infection has been found in O. flexuosellus. Thus, sex ratio bias might be linked to Wolbachia-infection in E. submarginalis, but not in O. flexuosellus.}, } @article {pmid32726353, year = {2020}, author = {Fraser, JE and O'Donnell, TB and Duyvestyn, JM and O'Neill, SL and Simmons, CP and Flores, HA}, title = {Novel phenotype of Wolbachia strain wPip in Aedes aegypti challenges assumptions on mechanisms of Wolbachia-mediated dengue virus inhibition.}, journal = {PLoS pathogens}, volume = {16}, number = {7}, pages = {e1008410}, pmid = {32726353}, issn = {1553-7374}, support = {102591/Z/13/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; Dengue/prevention & control/transmission ; *Dengue Virus ; Gram-Negative Bacterial Infections ; Microbial Interactions/*physiology ; Mosquito Vectors/*microbiology ; Pest Control, Biological/methods ; Phenotype ; *Wolbachia ; }, abstract = {The bacterial endosymbiont Wolbachia is a biocontrol tool that inhibits the ability of the Aedes aegypti mosquito to transmit positive-sense RNA viruses such as dengue and Zika. Growing evidence indicates that when Wolbachia strains wMel or wAlbB are introduced into local mosquito populations, human dengue incidence is reduced. Despite the success of this novel intervention, we still do not fully understand how Wolbachia protects mosquitoes from viral infection. Here, we demonstrate that the Wolbachia strain wPip does not inhibit virus infection in Ae. aegypti. We have leveraged this novel finding, and a panel of Ae. aegypti lines carrying virus-inhibitory (wMel and wAlbB) and non-inhibitory (wPip) strains in a common genetic background, to rigorously test a number of hypotheses about the mechanism of Wolbachia-mediated virus inhibition. We demonstrate that, contrary to previous suggestions, there is no association between a strain's ability to inhibit dengue infection in the mosquito and either its typical density in the midgut or salivary glands, or the degree to which it elevates innate immune response pathways in the mosquito. These findings, and the experimental platform provided by this panel of genetically comparable mosquito lines, clear the way for future investigations to define how Wolbachia prevents Ae. aegypti from transmitting viruses.}, } @article {pmid32722516, year = {2020}, author = {Chen, H and Zhang, M and Hochstrasser, M}, title = {The Biochemistry of Cytoplasmic Incompatibility Caused by Endosymbiotic Bacteria.}, journal = {Genes}, volume = {11}, number = {8}, pages = {}, pmid = {32722516}, issn = {2073-4425}, support = {R01 GM053756/GM/NIGMS NIH HHS/United States ; R35 GM136325/GM/NIGMS NIH HHS/United States ; GM046904 and GM053756/NH/NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/*microbiology/*pathology ; Drosophila melanogaster/*microbiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Many species of arthropods carry maternally inherited bacterial endosymbionts that can influence host sexual reproduction to benefit the bacterium. The most well-known of such reproductive parasites is Wolbachia pipientis. Wolbachia are obligate intracellular α-proteobacteria found in nearly half of all arthropod species. This success has been attributed in part to their ability to manipulate host reproduction to favor infected females. Cytoplasmic incompatibility (CI), a phenomenon wherein Wolbachia infection renders males sterile when they mate with uninfected females, but not infected females (the rescue mating), appears to be the most common. CI provides a reproductive advantage to infected females in the presence of a threshold level of infected males. The molecular mechanisms of CI and other reproductive manipulations, such as male killing, parthenogenesis, and feminization, have remained mysterious for many decades. It had been proposed by Werren more than two decades ago that CI is caused by a Wolbachia-mediated sperm modification and that rescue is achieved by a Wolbachia-encoded rescue factor in the infected egg. In the past few years, new research has highlighted a set of syntenic Wolbachia gene pairs encoding CI-inducing factors (Cifs) as the key players for the induction of CI and its rescue. Within each Cif pair, the protein encoded by the upstream gene is denoted A and the downstream gene B. To date, two types of Cifs have been characterized based on the enzymatic activity identified in the B protein of each protein pair; one type encodes a deubiquitylase (thus named CI-inducing deubiquitylase or cid), and a second type encodes a nuclease (named CI-inducing nuclease or cin). The CidA and CinA proteins bind tightly and specifically to their respective CidB and CinB partners. In transgenic Drosophila melanogaster, the expression of either the Cid or Cin protein pair in the male germline induces CI and the expression of the cognate A protein in females is sufficient for rescue. With the identity of the Wolbachia CI induction and rescue factors now known, research in the field has turned to directed studies on the molecular mechanisms of CI, which we review here.}, } @article {pmid32718933, year = {2020}, author = {Chung, M and Basting, PJ and Patkus, RS and Grote, A and Luck, AN and Ghedin, E and Slatko, BE and Michalski, M and Foster, JM and Bergman, CM and Hotopp, JCD}, title = {A Meta-Analysis of Wolbachia Transcriptomics Reveals a Stage-Specific Wolbachia Transcriptional Response Shared Across Different Hosts.}, journal = {G3 (Bethesda, Md.)}, volume = {10}, number = {9}, pages = {3243-3260}, pmid = {32718933}, issn = {2160-1836}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Filarioidea ; *Nematoda ; Symbiosis ; Transcriptome ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a genus containing obligate, intracellular endosymbionts with arthropod and nematode hosts. Numerous studies have identified differentially expressed transcripts in Wolbachia endosymbionts that potentially inform the biological interplay between these endosymbionts and their hosts, albeit with discordant results. Here, we re-analyze previously published Wolbachia RNA-Seq transcriptomics data sets using a single workflow consisting of the most up-to-date algorithms and techniques, with the aim of identifying trends or patterns in the pan-Wolbachia transcriptional response. We find that data from one of the early studies in filarial nematodes did not allow for robust conclusions about Wolbachia differential expression with these methods, suggesting the original interpretations should be reconsidered. Across datasets analyzed with this unified workflow, there is a general lack of global gene regulation with the exception of a weak transcriptional response resulting in the upregulation of ribosomal proteins in early larval stages. This weak response is observed across diverse Wolbachia strains from both nematode and insect hosts suggesting a potential pan-Wolbachia transcriptional response during host development that diverged more than 700 million years ago.}, } @article {pmid32712316, year = {2020}, author = {Taghikhani, R and Sharomi, O and Gumel, AB}, title = {Dynamics of a two-sex model for the population ecology of dengue mosquitoes in the presence of Wolbachia.}, journal = {Mathematical biosciences}, volume = {328}, number = {}, pages = {108426}, doi = {10.1016/j.mbs.2020.108426}, pmid = {32712316}, issn = {1879-3134}, mesh = {Aedes/*microbiology/*virology ; Animals ; Basic Reproduction Number/statistics & numerical data ; Computer Simulation ; Dengue/prevention & control/*transmission/virology ; Dengue Virus/pathogenicity ; Ecosystem ; Female ; Humans ; Male ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/*microbiology/*virology ; Pest Control, Biological/*methods/statistics & numerical data ; Population Dynamics ; Wolbachia/*physiology ; }, abstract = {The release of Wolbachia-infected mosquitoes into the population of wild mosquitoes is one of the promising biological control method for combating the population abundance of mosquitoes that cause deadly diseases, such as dengue. In this study, a new two-sex mathematical model for the population ecology of dengue mosquitoes and disease is designed and used to assess the population-level impact of the periodic release of Wolbachia-infected mosquitoes. Rigorous analysis of the model, which incorporates many of the lifecycle features of dengue disease and the cytoplasmic incompatibility property of Wolbachia bacterium in mosquitoes, reveal that the disease-free equilibrium of the model is locally-asymptotically stable whenever a certain epidemiological threshold, known as the reproduction number of the model (denoted by R0W), is less than unity. The model is shown, using centre manifold theory, to undergo the phenomenon of backward bifurcation at R0W=1. The consequence of this bifurcation is that Wolbachia may not persist, or dengue disease may not be effectively-controlled, when R0W is less than unity. Such persistence and elimination will depend on the initial sizes of the sub-populations of the model. Two mechanisms were identified for which the backward bifurcation phenomenon can be removed. When backward bifurcation does not occur, the associated non-trivial disease-free equilibrium is shown to be globally-asymptotically stable when the reproduction number of the model is less than unity. Numerical simulations, using data relevant to dengue transmission dynamics in northern Queensland, Australia, shows that releasing Wolbachia-infected mosquitoes every three weeks, for a one-year duration, can lead to the effective control of the population abundance of the local wild mosquitoes, and that such effective control increases with increasing number of Wolbachia-infected mosquitoes released (resulting in the reduction of over 90% of the wild mosquito population from their baseline values). Furthermore, simulations show that releasing only adult male Wolbachia-infected mosquitoes provide more beneficial population-level impact (in terms of reducing the population abundance of the wild mosquitoes), in comparison to releasing adult female Wolbachia-infected mosquitoes. Increasing the frequency of Wolbachia release (e.g., from the default release frequency of every three weeks to weekly) does not significantly affect the effectiveness of the Wolbachia-based control program in curtailing the local abundance of the wild mosquitoes. Finally, it was shown that the cytoplasmic incompatibility property of Wolbachia bacterium does not significantly affect the effectiveness of the Wolbachia-based mosquito control strategy implemented in the community.}, } @article {pmid32710084, year = {2020}, author = {Crawford, JE and Clarke, DW and Criswell, V and Desnoyer, M and Cornel, D and Deegan, B and Gong, K and Hopkins, KC and Howell, P and Hyde, JS and Livni, J and Behling, C and Benza, R and Chen, W and Dobson, KL and Eldershaw, C and Greeley, D and Han, Y and Hughes, B and Kakani, E and Karbowski, J and Kitchell, A and Lee, E and Lin, T and Liu, J and Lozano, M and MacDonald, W and Mains, JW and Metlitz, M and Mitchell, SN and Moore, D and Ohm, JR and Parkes, K and Porshnikoff, A and Robuck, C and Sheridan, M and Sobecki, R and Smith, P and Stevenson, J and Sullivan, J and Wasson, B and Weakley, AM and Wilhelm, M and Won, J and Yasunaga, A and Chan, WC and Holeman, J and Snoad, N and Upson, L and Zha, T and Dobson, SL and Mulligan, FS and Massaro, P and White, BJ}, title = {Author Correction: Efficient production of male Wolbachia-infected Aedes aegypti mosquitoes enables large-scale suppression of wild populations.}, journal = {Nature biotechnology}, volume = {38}, number = {8}, pages = {1000}, doi = {10.1038/s41587-020-0649-2}, pmid = {32710084}, issn = {1546-1696}, abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.}, } @article {pmid32708808, year = {2020}, author = {Lindsey, ARI}, title = {Sensing, Signaling, and Secretion: A Review and Analysis of Systems for Regulating Host Interaction in Wolbachia.}, journal = {Genes}, volume = {11}, number = {7}, pages = {}, pmid = {32708808}, issn = {2073-4425}, support = {R01AI144430//National Institute of Allergy and Infectious Diseases/International ; }, mesh = {Animals ; Arthropods/microbiology ; Female ; Gene Expression Regulation, Bacterial ; Genome, Bacterial/genetics ; Host-Pathogen Interactions/genetics ; Nematoda/microbiology ; Quorum Sensing/*genetics ; Secretory Pathway/*genetics ; Signal Transduction/genetics ; Symbiosis/*genetics ; Wolbachia/genetics/metabolism/*physiology ; }, abstract = {Wolbachia (Anaplasmataceae) is an endosymbiont of arthropods and nematodes that resides within host cells and is well known for manipulating host biology to facilitate transmission via the female germline. The effects Wolbachia has on host physiology, combined with reproductive manipulations, make this bacterium a promising candidate for use in biological- and vector-control. While it is becoming increasingly clear that Wolbachia's effects on host biology are numerous and vary according to the host and the environment, we know very little about the molecular mechanisms behind Wolbachia's interactions with its host. Here, I analyze 29 Wolbachia genomes for the presence of systems that are likely central to the ability of Wolbachia to respond to and interface with its host, including proteins for sensing, signaling, gene regulation, and secretion. Second, I review conditions under which Wolbachia alters gene expression in response to changes in its environment and discuss other instances where we might hypothesize Wolbachia to regulate gene expression. Findings will direct mechanistic investigations into gene regulation and host-interaction that will deepen our understanding of intracellular infections and enhance applied management efforts that leverage Wolbachia.}, } @article {pmid32708688, year = {2020}, author = {Krafsur, AM and Ghosh, A and Brelsfoard, CL}, title = {Phenotypic Response of Wolbachia pipientis in a Cell-Free Medium.}, journal = {Microorganisms}, volume = {8}, number = {7}, pages = {}, pmid = {32708688}, issn = {2076-2607}, abstract = {Wolbachia, an obligate intracellular bacterium estimated to infect millions of arthropod species worldwide, is currently being utilized in novel control strategies to limit the transmission of Dengue and Zika viruses. A limitation for Wolbachia-based control approaches is the difficulty of transferring Wolbachia to novel hosts and the lack of tools for the genetic transformation of Wolbachia due to the inability to culture Wolbachia outside the insect host cell in an axenic media. Here, we applied extracellular Wolbachia to phenotypic microarrays to measure the metabolic response of Wolbachia in media formulations with different pH levels and supplementation with Casamino acids. Results suggested a pH of 6.5-6.8 and showed that the supplementation of 1 mg/mL casamino acids increased the survival and longevity of Wolbachia in an axenic medium. In addition, phenotypic microarrays are a useful tool to measure the phenotypic response of Wolbachia under different media conditions, as well as determine specific components that may be required for an axenic medium. This study is an initial step toward the development of a potential Wolbachia axenic culture system.}, } @article {pmid32704304, year = {2020}, author = {Calamoneri, T and Gastaldello, M and Mary, A and Sagot, MF and Sinaimeri, B}, title = {Algorithms for the quantitative Lock/Key model of cytoplasmic incompatibility.}, journal = {Algorithms for molecular biology : AMB}, volume = {15}, number = {}, pages = {14}, pmid = {32704304}, issn = {1748-7188}, abstract = {Cytoplasmic incompatibility (CI) relates to the manipulation by the parasite Wolbachia of its host reproduction. Despite its widespread occurrence, the molecular basis of CI remains unclear and theoretical models have been proposed to understand the phenomenon. We consider in this paper the quantitative Lock-Key model which currently represents a good hypothesis that is consistent with the data available. CI is in this case modelled as the problem of covering the edges of a bipartite graph with the minimum number of chain subgraphs. This problem is already known to be NP-hard, and we provide an exponential algorithm with a non trivial complexity. It is frequent that depending on the dataset, there may be many optimal solutions which can be biologically quite different among them. To rely on a single optimal solution may therefore be problematic. To this purpose, we address the problem of enumerating (listing) all minimal chain subgraph covers of a bipartite graph and show that it can be solved in quasi-polynomial time. Interestingly, in order to solve the above problems, we considered also the problem of enumerating all the maximal chain subgraphs of a bipartite graph and improved on the current results in the literature for the latter. Finally, to demonstrate the usefulness of our methods we show an application on a real dataset.}, } @article {pmid32694524, year = {2020}, author = {Ren, W and Wei, H and Yang, Y and Shao, S and Wu, H and Chen, X and Yang, Z}, title = {Molecular detection and phylogenetic analyses of Wolbachia in natural populations of nine galling Aphid species.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {12025}, pmid = {32694524}, issn = {2045-2322}, mesh = {Animals ; Aphids/*microbiology ; Genes, Bacterial ; Genome, Bacterial ; *Phylogeny ; RNA, Bacterial ; RNA, Ribosomal, 16S/genetics ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia is one of the most abundant facultative intracellular symbionts in arthropods. It alters host biology in diverse ways, including the induction of reproductive manipulation, association of nutrient supplier and protection against pathogens. Aphids are a group of insects which exhibit interesting biological characteristics such as complex life cycles, alteration of sexual and asexual reproduction and shifts between two different hosts. Wolbachia is widely present in many orders of insects, but so far limited studies on Wolbachia in aphids have been carried out. Galling aphids are a group of aphids that induce galls on their primary host plants at specific life stage. In this study, 15 natural populations representing nine galling aphid species were analyzed for the presence of Wolbachia using species-specific primer pairs. Wolbachia presence in galling aphids was quite low and varied significantly among aphid populations. Only three of the 15 populations we analyzed had detectable Wolbachia and the overall infection rate was 20%. Two Wolbachia strains, O and B, were identified from the galling aphids Kaburagia rhusicola and Schlechtendalia chinensis. Strain O was for the first time to be found in aphids, and it is likely involved with the life stages of galling aphids living in closed microenvironments with specific survival strategies that are different from free-living aphids.}, } @article {pmid32690078, year = {2020}, author = {Huang, EYY and Wong, AYP and Lee, IHT and Qu, Z and Yip, HY and Leung, CW and Yin, SM and Hui, JHL}, title = {Infection patterns of dengue, Zika and endosymbiont Wolbachia in the mosquito Aedes albopictus in Hong Kong.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {361}, pmid = {32690078}, issn = {1756-3305}, mesh = {*Aedes/microbiology/virology ; Animals ; Dengue/transmission ; Dengue Virus/*isolation & purification ; Hong Kong/epidemiology ; Mosquito Vectors/virology ; Pathology, Molecular ; Polymerase Chain Reaction ; *Prevalence ; Symbiosis ; Wolbachia/*isolation & purification ; Zika Virus/*isolation & purification ; Zika Virus Infection/transmission ; }, abstract = {BACKGROUND: The mosquito Aedes albopictus is a vector of dengue and Zika viruses. Insecticide-resistant mosquito populations have evolved in recent decades, suggesting that new control strategies are needed. Hong Kong has a monsoon-influenced humid subtropical climate, which favours the spread of mosquitoes. However, baseline information on the composition and dynamics of the occurrence of endosymbiont Wolbachia in local Ae. albopictus is lacking, hindering the development of scientifically-informed control measures. This study identifies the presence and absence of dengue and Zika viruses, and Wolbachia infection in Aedes albopictus in Hong Kong.

METHODS: Oviposition traps were set at 57 areas in Hong Kong, and both immature and adult mosquitoes were collected on a monthly basis between April 2018 and April 2019 as the study sample. Each individual mosquito in this sample was processed and screened for the presence of the dengue and Zika viruses and the endosymbionts Wolbachia wAlbA and wAlbB with PCR.

RESULTS: Totals of 967 and 984 mosquitoes were tested respectively for the presence of dengue and Zika viruses, and no trace of either infection was found in these samples. The presence of wAlbA and wAlbB was also tested in 1582 individuals. Over 80% of these individuals were found to be stably infected with Wolbachia throughout the thirteen-month collection period (~ 47% singly-infected; ~ 36.8% doubly infected with both wAlbA and wAlbB).

CONCLUSIONS: The high degree of Wolbachia wAlbA and wAlbB infection in Ae. albopictus mosquitoes in Hong Kong, coupled with the absence of any signs of infection by dengue and Zika viruses, contrasts significantly with the pattern of mosquito infection in other parts of Asia. Further studies of the infection pattern in local mosquitoes are warranted before mosquito control strategies used in other regions are implemented in Hong Kong.}, } @article {pmid32689950, year = {2020}, author = {Morrow, JL and Om, N and Beattie, GAC and Chambers, GA and Donovan, NJ and Liefting, LW and Riegler, M and Holford, P}, title = {Characterization of the bacterial communities of psyllids associated with Rutaceae in Bhutan by high throughput sequencing.}, journal = {BMC microbiology}, volume = {20}, number = {1}, pages = {215}, pmid = {32689950}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; Bhutan ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Hemiptera/*microbiology ; High-Throughput Nucleotide Sequencing ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Rutaceae/microbiology/*parasitology ; Sequence Analysis, DNA/*methods ; }, abstract = {BACKGROUND: Several plant-pathogenic bacteria are transmitted by insect vector species that often also act as hosts. In this interface, these bacteria encounter plant endophytic, insect endosymbiotic and other microbes. Here, we used high throughput sequencing to examine the bacterial communities of five different psyllids associated with citrus and related plants of Rutaceae in Bhutan: Diaphorina citri, Diaphorina communis, Cornopsylla rotundiconis, Cacopsylla heterogena and an unidentified Cacopsylla sp.

RESULTS: The microbiomes of the psyllids largely comprised their obligate P-endosymbiont 'Candidatus Carsonella ruddii', and one or two S-endosymbionts that are fixed and specific to each lineage. In addition, all contained Wolbachia strains; the Bhutanese accessions of D. citri were dominated by a Wolbachia strain first found in American isolates of D. citri, while D. communis accessions were dominated by the Wolbachia strain, wDi, first detected in D. citri from China. The S-endosymbionts from the five psyllids grouped with those from other psyllid taxa; all D. citri and D. communis individuals contained sequences matching 'Candidatus Profftella armatura' that has previously only been reported from other Diaphorina species, and the remaining psyllid species contained OTUs related to unclassified Enterobacteriaceae. The plant pathogenic 'Candidatus Liberibacter asiaticus' was found in D. citri but not in D. communis. Furthermore, an unidentified 'Candidatus Liberibacter sp.' occurred at low abundance in both Co. rotundiconis and the unidentified Cacopsylla sp. sampled from Zanthoxylum sp.; the status of this new liberibacter as a plant pathogen and its potential plant hosts are currently unknown. The bacterial communities of Co. rotundiconis also contained a range of OTUs with similarities to bacteria previously found in samples taken from various environmental sources.

CONCLUSIONS: The bacterial microbiota detected in these Bhutanese psyllids support the trends that have been seen in previous studies: psyllids have microbiomes largely comprising their obligate P-endosymbiont and one or two S-endosymbionts. In addition, the association with plant pathogens has been demonstrated, with the detection of liberibacters in a known host, D. citri, and identification of a putative new species of liberibacter in Co. rotundiconis and Cacopsylla sp.}, } @article {pmid32649049, year = {2020}, author = {Nguyen, HM and Theppannga, W and Vongphayloth, K and Douangngeun, B and Blacksell, SD and Robinson, MT}, title = {Screening of ectoparasites from domesticated dogs for bacterial pathogens in Vientiane, Lao PDR.}, journal = {Zoonoses and public health}, volume = {67}, number = {8}, pages = {862-868}, pmid = {32649049}, issn = {1863-2378}, support = {106698/Z/14/Z//Wellcome/ ; }, mesh = {Animals ; Bacteria/*isolation & purification ; Bacterial Infections/epidemiology/microbiology/*veterinary ; Ctenocephalides/*microbiology ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Female ; Flea Infestations/epidemiology/veterinary ; Laos/epidemiology ; Lice Infestations/epidemiology/veterinary ; Male ; Phthiraptera/classification/*microbiology ; Rhipicephalus sanguineus/*microbiology ; Tick Infestations/epidemiology/veterinary ; }, abstract = {Arthropod-borne diseases are widespread worldwide and are a complex interaction between animals, humans and ectoparasites. The understanding of the diversity and epidemiology of organisms transmitted by arthropod vectors, and the role of hosts and vectors in transmission of infections remain limited in Lao PDR. What knowledge does exist is primarily focused on more rural regions of the country. This study screened ectoparasites from domestic dogs in Vientiane city for the presence of bacterial pathogens of zoonotic importance. A total of 3,511 arthropod vectors were collected from 112 dogs. Vectors collected were Rhipicephalus sanguineus ticks, Ctenocephalides felis felis and Ctenocephalides felis orientis fleas and Heterodoxus spiniger lice. A sub-sample of vectors from each dog was analysed by PCR to identify the potential bacteria. From 129 vector pools, Rickettsia spp. was detected in 6.7% (7/105) pools of ticks, 86.4% (19/22) pools of fleas and both pools of lice. Sequencing analysis confirmed Rickettsia felis in 13 flea pools and one louse pool and Rickettsia asembonensis in six flea pools. Anaplasmataceae was identified in 14.3% (15/105) tick pools and 100% (22/22) flea pools. Sequencing revealed the presence of Anaplasma platys in ticks and Wolbachia pipientis in fleas. Leptospira spp. was detected in one tick and one louse pool, and Brucella spp. was detected in 12.4% (13/105) tick pools. All samples were negative for Bartonella spp., Coxiella burnetii and Borrelia burgdorferi. This is the first study providing evidence of R. asembonensis in fleas in Laos. Results from this study show arthropods are potential vectors to transmit zoonotic infection in Vientiane city, suggesting humans are at risk of zoonotic infections in the city.}, } @article {pmid32647904, year = {2020}, author = {Félix, ML and Armúa-Fernández, MT and Parodi, P and Bazzano, V and Mangold, AJ and Venzal, JM}, title = {Detection of a putative novel genotype of Anaplasma in gray-brocket deer (Mazama gouazoubira) from Uruguay.}, journal = {Experimental & applied acarology}, volume = {81}, number = {4}, pages = {575-583}, doi = {10.1007/s10493-020-00523-0}, pmid = {32647904}, issn = {1572-9702}, support = {Programa Iniciación a la Investigación 2017, Project ID 160//Comisión Sectorial de Investigación Científica/ ; }, mesh = {*Anaplasma/genetics ; Animals ; Deer/*microbiology ; Genotype ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Uruguay ; }, abstract = {Anaplasmataceae includes the genera Anaplasma, Ehrlichia, Neorickettsia and Wolbachia, comprising a group of obligate intracellular bacteria. The genus Anaplasma has pathogenic species transmitted by ticks of veterinary and human health importance. Wild ungulates such as deer represent important reservoirs and amplifiers of Anaplasmataceae. The interaction between deer and domestic ruminants represents a serious problem due to the transmission of these pathogens through their ectoparasites. In the present study, we investigated the presence of Anaplasmataceae organisms in blood, tissues and tick samples of a gray-brocket deer (Mazama gouazoubira). The specimen was found dead in a farm in northeast Uruguay. PCRs targeting partial regions of 16S rRNA and groESL genes were carried out for Anaplasmataceae DNA detection. Moreover, several ectoparasites were identified: the chewing louse Tricholipeurus albimarginatus, the Neotropical deer louse fly Lipoptena mazamae, and the ticks Haemaphysalis juxtakochi and Rhipicephalus microplus. A consensus sequence of 1274 bp of 16S rRNA was generated for Anaplasma sp. from the M. gouazoubira blood sample. All ticks analysed by PCR assays were negative. No band was detected in any of the samples after PCR targeting groESL gene. Phylogenetic analysis using 16S rRNA partial gene sequences, clustered the putative novel genotype sequence obtained in this study, named Anaplasma sp. genotype Mazama-Uruguay, along with Anaplasma sp. detected in Mazama sp., Mazama americana and Mazama bororo, all deer species from Brazil. Furthermore, this cluster showed to be closely related to Anaplasma bovis sequences obtained from various ruminants and other mammals from several parts of the world. The pathogenicity as well as its infecting potential to other cervids or domestic ruminants is currently unknown. Further studies should be performed in order to characterize this novel species, especially targeting other genes.}, } @article {pmid32641039, year = {2020}, author = {Brady, OJ and Kharisma, DD and Wilastonegoro, NN and O'Reilly, KM and Hendrickx, E and Bastos, LS and Yakob, L and Shepard, DS}, title = {The cost-effectiveness of controlling dengue in Indonesia using wMel Wolbachia released at scale: a modelling study.}, journal = {BMC medicine}, volume = {18}, number = {1}, pages = {186}, pmid = {32641039}, issn = {1741-7015}, support = {206471/Z/17/Z/WT_/Wellcome Trust/United Kingdom ; OPP1187889//Bill and Melinda Gates Foundation/International ; 206471/Z/17/Z//Wellcome Trust (GB)/International ; OP1183567//Bill and Melinda Gates Foundation/International ; }, mesh = {Animals ; Cost-Benefit Analysis/*methods ; Dengue/*economics/epidemiology/*therapy ; Humans ; Indonesia/epidemiology ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: Release of virus-blocking Wolbachia-infected mosquitoes is an emerging disease control strategy that aims to control dengue and other arboviral infections. Early entomological data and modelling analyses have suggested promising outcomes, and wMel Wolbachia releases are now ongoing or planned in 12 countries. To help inform government, donor, or philanthropist decisions on scale-up beyond single city releases, we assessed this technology's cost-effectiveness under alternative programmatic options.

METHODS: Using costing data from existing Wolbachia releases, previous dynamic model-based estimates of Wolbachia effectiveness, and a spatially explicit model of release and surveillance requirements, we predicted the costs and effectiveness of the ongoing programme in Yogyakarta City and three new hypothetical programmes in Yogyakarta Special Autonomous Region, Jakarta, and Bali.

RESULTS: We predicted Wolbachia to be a highly cost-effective intervention when deployed in high-density urban areas with gross cost-effectiveness below $1500 per DALY averted. When offsets from the health system and societal perspective were included, such programmes even became cost saving over 10-year time horizons with favourable benefit-cost ratios of 1.35 to 3.40. Sequencing Wolbachia releases over 10 years could reduce programme costs by approximately 38% compared to simultaneous releases everywhere, but also delays the benefits. Even if unexpected challenges occurred during deployment, such as emergence of resistance in the medium-term or low effective coverage, Wolbachia would remain a cost-saving intervention.

CONCLUSIONS: Wolbachia releases in high-density urban areas are expected to be highly cost-effective and could potentially be the first cost-saving intervention for dengue. Sites with strong public health infrastructure, fiscal capacity, and community support should be prioritised.}, } @article {pmid32640111, year = {2022}, author = {Liu, G and Tan, FH and Lau, SA and Jaafar, MH and Chung, FY and Azzam, G and Liong, MT and Li, Y}, title = {Lactic acid bacteria feeding reversed the malformed eye structures and ameliorated gut microbiota profiles of Drosophila melanogaster Alzheimer's disease model.}, journal = {Journal of applied microbiology}, volume = {132}, number = {4}, pages = {3155-3167}, doi = {10.1111/jam.14773}, pmid = {32640111}, issn = {1365-2672}, support = {1001/PTEKIND/870030//USM-RIKEN International Centre for Aging Science (URICAS)/ ; //Centre of Excellence for Biotechnology, Chinese Academy of Sciences-The Third World Academy of Sciences/ ; }, mesh = {*Acetobacter ; *Alzheimer Disease/genetics/pathology ; Animals ; Drosophila melanogaster/genetics/microbiology ; *Gastrointestinal Microbiome ; *Lactobacillales ; }, abstract = {AIMS: To utilize transgenic GMR-Aβ42 Drosophila melanogaster as a model to evaluate potential Alzheimer's disease (AD)-reversal effects via the administration of lactic acid bacteria (LAB) strains, and associations of LAB with changes in gut microbiota profiles.

METHODS AND RESULTS: Wild-type flies (Oregon-R) were crossed with glass multimer reporter-GAL4 (GMR-GAL4) to produce GMR-OreR (Control), while UAS-Aβ42 (#33769) were crossed with GMR-GAL4 to produce transgenic Drosophila line that expressed Aβ42 (GMR-Aβ42). Feed containing seven different LAB strains (Lactobacillus paracasei 0291, Lactobacillus helveticus 1515, Lactobacillus reuteri 30242, L. reuteri 8513d, Lactobacillus fermentum 8312, Lactobacillus casei Y, Lactobacillus sakei Probio65) were given to GMR-Aβ42 respectively, while feed without LAB strains were given to control and transgenic GMR-Aβ42.nf Drosophila lines. The morphology of the eyes was viewed with scanning electron microscopy (SEM). The changes in gut microbiota profiles associated with LAB were analysed using 16s high throughput sequencing. Malformation of eye structures in transgenic GMR-Aβ42 Drosophila were reversed upon the administration of LAB strains, with more prevalent effects from L. sakei Probio65 and L. paracasei 0291. The GMR-Aβ42.nf group showed dominance of Wolbachia in the gut, a genus that was almost absent in the normal control group (P < 0·05). The administration of L. sakei Probio65 and L. paracasei 0291 reduced the abundance of Wolbachia accompanied by increased abundance of Stenotrophomonas and Acetobacter (P < 0·05), resembling the microbial profile of the control group.

CONCLUSIONS: Lactobacillus sakei Probio65 and Lactobacillus paracasei 0291 have more prominent effects in reversing malformed eye of transgenic GMR-Aβ42 Drosophila, and reducing the abundance of Wolbachia accompanied by an increased abundance of Stenotrophomonas and Acetobacter.

Potentials of LAB to prevent and/or alleviate the onset and pathogenesis of neurodegenerative diseases such as AD, supporting brain health strategies along the gut-brain axis.}, } @article {pmid32639986, year = {2020}, author = {Gunderson, EL and Vogel, I and Chappell, L and Bulman, CA and Lim, KC and Luo, M and Whitman, JD and Franklin, C and Choi, YJ and Lefoulon, E and Clark, T and Beerntsen, B and Slatko, B and Mitreva, M and Sullivan, W and Sakanari, JA}, title = {The endosymbiont Wolbachia rebounds following antibiotic treatment.}, journal = {PLoS pathogens}, volume = {16}, number = {7}, pages = {e1008623}, pmid = {32639986}, issn = {1553-7374}, mesh = {Animals ; Brugia pahangi/*microbiology ; Female ; Filariasis/*microbiology ; Filaricides/*pharmacology ; Gerbillinae ; Rifampin/*pharmacology ; Wolbachia/*drug effects ; }, abstract = {Antibiotic treatment has emerged as a promising strategy to sterilize and kill filarial nematodes due to their dependence on their endosymbiotic bacteria, Wolbachia. Several studies have shown that novel and FDA-approved antibiotics are efficacious at depleting the filarial nematodes of their endosymbiont, thus reducing female fecundity. However, it remains unclear if antibiotics can permanently deplete Wolbachia and cause sterility for the lifespan of the adult worms. Concerns about resistance arising from mass drug administration necessitate a careful exploration of potential Wolbachia recrudescence. In the present study, we investigated the long-term effects of the FDA-approved antibiotic, rifampicin, in the Brugia pahangi jird model of infection. Initially, rifampicin treatment depleted Wolbachia in adult worms and simultaneously impaired female worm fecundity. However, during an 8-month washout period, Wolbachia titers rebounded and embryogenesis returned to normal. Genome sequence analyses of Wolbachia revealed that despite the population bottleneck and recovery, no genetic changes occurred that could account for the rebound. Clusters of densely packed Wolbachia within the worm's ovarian tissues were observed by confocal microscopy and remained in worms treated with rifampicin, suggesting that they may serve as privileged sites that allow Wolbachia to persist in worms while treated with antibiotic. To our knowledge, these clusters have not been previously described and may be the source of the Wolbachia rebound.}, } @article {pmid32635864, year = {2020}, author = {Rosenwald, LC and Sitvarin, MI and White, JA}, title = {Endosymbiotic Rickettsiella causes cytoplasmic incompatibility in a spider host.}, journal = {Proceedings. Biological sciences}, volume = {287}, number = {1930}, pages = {20201107}, pmid = {32635864}, issn = {1471-2954}, mesh = {Animals ; Coxiellaceae/*physiology ; Cytoplasm/*microbiology ; Female ; Host-Pathogen Interactions ; Male ; Spiders/*microbiology ; Symbiosis ; }, abstract = {Many arthropod hosts are infected with bacterial endosymbionts that manipulate host reproduction, but few bacterial taxa have been shown to cause such manipulations. Here, we show that a bacterial strain in the genus Rickettsiella causes cytoplasmic incompatibility (CI) between infected and uninfected hosts. We first surveyed the bacterial community of the agricultural spider Mermessus fradeorum (Linyphiidae) using high throughput sequencing and found that individual spiders can be infected with up to five different strains of maternally inherited symbiont from the genera Wolbachia, Rickettsia, and Rickettsiella. The Rickettsiella strain was pervasive, found in all 23 tested spider matrilines. We used antibiotic curing to generate uninfected matrilines that we reciprocally crossed with individuals infected only with Rickettsiella. We found that only 13% of eggs hatched when uninfected females were mated with Rickettsiella-infected males; in contrast, at least 83% of eggs hatched in the other cross types. This is the first documentation of Rickettsiella, or any Gammaproteobacteria, causing CI. We speculate that induction of CI may be much more widespread among maternally inherited bacteria than previously appreciated. Further, our results reinforce the importance of thoroughly characterizing and assessing the inherited microbiome before attributing observed host phenotypes to well-characterized symbionts such as Wolbachia.}, } @article {pmid32631404, year = {2020}, author = {Faria Waziry, PA and Raja, A and Salmon, C and Aldana, N and Damodar, S and Fukushima, AR and Mayi, BS}, title = {Impact of pyriproxyfen on virus behavior: implications for pesticide-induced virulence and mechanism of transmission.}, journal = {Virology journal}, volume = {17}, number = {1}, pages = {93}, pmid = {32631404}, issn = {1743-422X}, mesh = {Aedes/virology ; Animals ; Brazil ; Cell Survival/drug effects ; Dengue Virus/drug effects ; Extracellular Vesicles/drug effects/virology ; Flavivirus Infections/*transmission ; Green Fluorescent Proteins/genetics ; HeLa Cells ; Humans ; Insecticides/*pharmacology ; Jurkat Cells ; Pyridines/*pharmacology ; Trophoblasts/drug effects/virology ; Vesiculovirus/*drug effects ; Virulence ; Virus Replication/*drug effects ; Zika Virus/drug effects ; }, abstract = {BACKGROUND: More than 3 years since the last Zika virus (ZIKV) outbreak in Brazil, researchers are still deciphering the molecular mechanisms of neurovirulence and vertical transmission, as well as the best way to control spread of ZIKV, a flavivirus. The use of pesticides was the main strategy of mosquito control during the last ZIKV outbreak.

METHODS: We used vesicular stomatitis virus (VSV) tagged with green fluorescent protein (GFP) as our prototypical virus to study the impact of insecticide pyriproxyfen (PPF). VZV-GFP infected and uninfected Jurkat, HeLa and trophoblast cells were treated with PPF and compared to untreated cells (control). Cell viability was determined by the MTT assay. Cell morphology, presence of extracellular vesicles (EVs), virus infection/GFP expression as well as active mitochondrial levels/localization were examined by confocal microscopy.

RESULTS: PPF, which was used to control mosquito populations in Brazil prior to the ZIKV outbreak, enhances VSV replication and has cell membrane-altering properties in the presence of virus. PPF causes enhanced viral replication and formation of large EVs, loaded with virus as well as mitochondria. Treatment of trophoblasts or HeLa cells with increasing concentrations of PPF does not alter cell viability, however, it proportionately increases Jurkat cell viability. Increasing concentrations of PPF followed by VSV infection does not interfere with HeLa cell viability. Both Jurkats and trophoblasts show proportionately increased cell death with increased concentrations of PPF in the presence of virus.

CONCLUSIONS: We hypothesize that PPF disrupts the lipid microenvironment of mammalian cells, thereby interfering with pathways of viral replication. PPF lowers viability of trophoblasts and Jurkats in the presence of VSV, implying that the combination renders immune system impairment in infected individuals as well as enhanced vulnerability of fetuses towards viral vertical transmission. We hypothesize that similar viruses such as ZIKV may be vertically transmitted via EV-to-cell contact when exposed to PPF, thereby bypassing immune detection. The impact of pesticides on viral replication must be fully investigated before large scale use in future outbreaks of mosquito borne viruses.}, } @article {pmid32630209, year = {2020}, author = {Khoo, JJ and Kurtti, TJ and Husin, NA and Beliavskaia, A and Lim, FS and Zulkifli, MMS and Al-Khafaji, AM and Hartley, C and Darby, AC and Hughes, GL and AbuBakar, S and Makepeace, BL and Bell-Sakyi, L}, title = {Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines.}, journal = {Microorganisms}, volume = {8}, number = {7}, pages = {}, pmid = {32630209}, issn = {2076-2607}, support = {MO002-2019//Ministry of Education, Malaysia/ ; 332192305//Malaysian Industry - Government Group for High Technology/ ; R21 AI138074/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; BB/P024378/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 332192305//Department for Business, Energy and Industrial Strategy, UK Government/ ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/T001240/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, abstract = {Wolbachia are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although Wolbachia DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not Wolbachia can infect and grow in tick cells, cell lines from three tick species, Ixodes scapularis, Ixodes ricinus and Rhipicephalus microplus, were inoculated with Wolbachia strains wStri and wAlbB isolated from mosquito cell lines. Homogenates prepared from fleas collected from cats in Malaysia were inoculated into an I. scapularis cell line. Bacterial growth and identity were monitored by microscopy and PCR amplification and sequencing of fragments of Wolbachia genes. The wStri strain infected Ixodes spp. cells and was maintained through 29 passages. The wAlbB strain successfully infected Ixodes spp. and R. microplus cells and was maintained through 2-5 passages. A novel strain of Wolbachia belonging to the supergroup F, designated wCfeF, was isolated in I. scapularis cells from a pool of Ctenocephalides sp. cat fleas and maintained in vitro through two passages over nine months. This is the first confirmed isolation of a Wolbachia strain from a flea and the first isolation of any Wolbachia strain outside the "pandemic" A and B supergroups. The study demonstrates that tick cells can host multiple Wolbachia strains, and can be added to panels of insect cell lines to improve success rates in isolation of field strains of Wolbachia.}, } @article {pmid32616636, year = {2020}, author = {Lebov, JF and Mattick, J and Libro, S and Sparklin, BC and Chung, M and Bromley, RE and Nadendla, S and Zhao, X and Ott, S and Sadzewicz, L and Tallon, LJ and Michalski, ML and Foster, JM and Dunning Hotopp, JC}, title = {Complete Genome Sequence of wBp, the Wolbachia Endosymbiont of Brugia pahangi FR3.}, journal = {Microbiology resource announcements}, volume = {9}, number = {27}, pages = {}, pmid = {32616636}, issn = {2576-098X}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {Lymphatic filariasis is a devastating disease caused by filarial nematode roundworms, which contain obligate Wolbachia endosymbionts. Here, we assembled the genome of wBp, the Wolbachia endosymbiont of the filarial nematode Brugia pahangi, from Illumina, Pacific Biosciences, and Oxford Nanopore data. The complete, circular genome is 1,072,967 bp.}, } @article {pmid32616041, year = {2020}, author = {Duan, XZ and Sun, JT and Wang, LT and Shu, XH and Guo, Y and Keiichiro, M and Zhu, YX and Bing, XL and Hoffmann, AA and Hong, XY}, title = {Recent infection by Wolbachia alters microbial communities in wild Laodelphax striatellus populations.}, journal = {Microbiome}, volume = {8}, number = {1}, pages = {104}, pmid = {32616041}, issn = {2049-2618}, support = {31672035 and 31871976//National Natural Science Foundation of China/International ; }, mesh = {Animals ; China ; Hemiptera/*microbiology ; Japan ; *Microbiota/genetics ; RNA, Ribosomal, 16S/genetics ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Host-associated microbial communities play an important role in the fitness of insect hosts. However, the factors shaping microbial communities in wild populations, including genetic background, ecological factors, and interactions among microbial species, remain largely unknown.

RESULTS: Here, we surveyed microbial communities of the small brown planthopper (SBPH, Laodelphax striatellus) across 17 geographical populations in China and Japan by using 16S rRNA amplicon sequencing. Using structural equation models (SEM) and Mantel analyses, we show that variation in microbial community structure is likely associated with longitude, annual mean precipitation (Bio12), and mitochondrial DNA variation. However, a Wolbachia infection, which is spreading to northern populations of SBPH, seems to have a relatively greater role than abiotic factors in shaping microbial community structure, leading to sharp decreases in bacterial taxon diversity and abundance in host-associated microbial communities. Comparative RNA-Seq analyses between Wolbachia-infected and -uninfected strains indicate that the Wolbachia do not seem to alter the immune reaction of SBPH, although Wolbachia affected expression of metabolism genes.

CONCLUSION: Together, our results identify potential factors and interactions among different microbial species in the microbial communities of SBPH, which can have effects on insect physiology, ecology, and evolution. Video Abstract.}, } @article {pmid32605600, year = {2020}, author = {Lefoulon, E and Clark, T and Borveto, F and Perriat-Sanguinet, M and Moulia, C and Slatko, BE and Gavotte, L}, title = {Pseudoscorpion Wolbachia symbionts: diversity and evidence for a new supergroup S.}, journal = {BMC microbiology}, volume = {20}, number = {1}, pages = {188}, pmid = {32605600}, issn = {1471-2180}, support = {-//New England Biolabs/International ; }, mesh = {Animals ; Arachnida/*microbiology ; Biotin/*genetics ; Gene Transfer, Horizontal ; Genome Size ; Genome, Bacterial ; Molecular Sequence Annotation ; Multilocus Sequence Typing ; Operon ; Phylogeny ; Symbiosis ; Whole Genome Sequencing/*methods ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia are the most widely spread endosymbiotic bacteria, present in a wide variety of insects and two families of nematodes. As of now, however, relatively little genomic data has been available. The Wolbachia symbiont can be parasitic, as described for many arthropod systems, an obligate mutualist, as in filarial nematodes or a combination of both in some organisms. They are currently classified into 16 monophyletic lineage groups ("supergroups"). Although the nature of these symbioses remains largely unknown, expanded Wolbachia genomic data will contribute to understanding their diverse symbiotic mechanisms and evolution.

RESULTS: This report focuses on Wolbachia infections in three pseudoscorpion species infected by two distinct groups of Wolbachia strains, based upon multi-locus phylogenies. Geogarypus minor harbours wGmin and Chthonius ischnocheles harbours wCisc, both closely related to supergroup H, while Atemnus politus harbours wApol, a member of a novel supergroup S along with Wolbachia from the pseudoscorpion Cordylochernes scorpioides (wCsco). Wolbachia supergroup S is most closely related to Wolbachia supergroups C and F. Using target enrichment by hybridization with Wolbachia-specific biotinylated probes to capture large fragments of Wolbachia DNA, we produced two draft genomes of wApol. Annotation of wApol highlights presence of a biotin operon, which is incomplete in many sequenced Wolbachia genomes.

CONCLUSIONS: The present study highlights at least two symbiont acquisition events among pseudoscorpion species. Phylogenomic analysis indicates that the Wolbachia from Atemnus politus (wApol), forms a separate supergroup ("S") with the Wolbachia from Cordylochernes scorpioides (wCsco). Interestingly, the biotin operon, present in wApol, appears to have been horizontally transferred multiple times along Wolbachia evolutionary history.}, } @article {pmid32604846, year = {2020}, author = {Kirik, H and Tummeleht, L and Lilja, T and Kurina, O}, title = {Novel Mitochondrial DNA Lineage Found among Ochlerotatus communis (De Geer, 1776) of the Nordic-Baltic Region.}, journal = {Insects}, volume = {11}, number = {6}, pages = {}, pmid = {32604846}, issn = {2075-4450}, support = {IUT21-1//Eesti Teadusagentuur/ ; 8P160014VLVP//Eesti Maaülikool/ ; }, abstract = {The Ochlerotatus (Oc.) communis complex consist of three Northern American species as well as a common Holarctic mosquito (Diptera: Culicidae) Oc. communis (De Geer, 1776). These sister species exhibit important ecological differences and are capable of transmitting various pathogens, but cannot always be differentiated by morphological traits. To investigate the Oc. communis complex in Europe, we compared three molecular markers (COI, ND5 and ITS2) from 54 Estonian mosquitoes as well as two COI marker sequences from Sweden. These sequences were subjected to phylogenetic analysis and screened for Wolbachia Hertig and Wolbach symbionts. Within and between groups, distances were calculated for each marker to better understand the relationships among individuals. Results demonstrate that a group of samples, extracted from adult female mosquitoes matching the morphology of Oc. communis, show a marked difference from the main species when comparing the mitochondrial markers COI and ND5. However, there is no variance between the same specimens when considering the nuclear ITS2. We conclude that Oc. communis encompasses two distinct mitochondrial DNA lineages in the Nordic-Baltic region. Further research is needed to investigate the origin and extent of these genetic differences.}, } @article {pmid32601334, year = {2020}, author = {Herran, B and Geniez, S and Delaunay, C and Raimond, M and Lesobre, J and Bertaux, J and Slatko, B and Grève, P}, title = {The shutting down of the insulin pathway: a developmental window for Wolbachia load and feminization.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {10551}, pmid = {32601334}, issn = {2045-2322}, mesh = {Animals ; Feminization/*metabolism ; Insulin/*metabolism ; Insulin Resistance/*physiology ; Isopoda/*metabolism ; Male ; Signal Transduction/physiology ; Wolbachia ; }, abstract = {Using the isopod Armadillidium vulgare as a case study, we review the significance of the "bacterial dosage model", which connects the expression of the extended phenotype to the rise of the Wolbachia load. In isopods, the Insulin-like Androgenic Gland hormone (IAG) induces male differentiation: Wolbachia feminizes males through insulin resistance, presumably through defunct insulin receptors. This should prevent an autocrine development of the androgenic glands so that females differentiate instead: feminization should translate as IAG silencing and increased Wolbachia load in the same developmental window. In line with the autocrine model, uninfected males expressed IAG from the first larval stage on, long before the androgenic gland primordia begin to differentiate, and exponentially throughout development. In contrast in infected males, expression fully stopped at stage 4 (juvenile), when male differentiation begins. This co-occurred with the only significant rise in the Wolbachia load throughout the life-stages. Concurrently, the raw expression of the bacterial Secretion Systems co-increased, but they were not over-expressed relative to the number of bacteria. The isopod model leads to formulate the "bacterial dosage model" throughout extended phenotypes as the conjunction between bacterial load as the mode of action, timing of multiplication (pre/post-zygotic), and site of action (soma vs. germen).}, } @article {pmid32599512, year = {2020}, author = {Ferreira, AG and Fairlie, S and Moreira, LA}, title = {Insect vectors endosymbionts as solutions against diseases.}, journal = {Current opinion in insect science}, volume = {40}, number = {}, pages = {56-61}, doi = {10.1016/j.cois.2020.05.014}, pmid = {32599512}, issn = {2214-5753}, mesh = {Animals ; Communicable Disease Control/*methods ; Disease Transmission, Infectious/*prevention & control ; Humans ; Insect Vectors/*physiology ; Insecta/*physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Viral diseases transmitted by mosquitoes, known as arboviruses, pose a significant threat to human life and are a major burden on many health systems around the world. Currently, arbovirus control strategies rely on insecticides or vector source reduction and, in the absence of effective, accessible and affordable vaccines, mainly on symptomatic based, non-specific treatments. However, insecticides have the potential to interfere with non-target organisms, cause environmental toxicity and insecticide resistance reduces their effectiveness as a sustainable control method. Complementary and sustainable strategies are urgently needed. Wolbachia, an invertebrate endosymbiont, has been used as an alternative strategy for arboviral control, through suppression or modification of mosquito populations. Here we discuss the burden that arboviruses impose on human populations and how Wolbachia can be used as a sustainable strategy for control, in alignment with the United Nations- 2030 Agenda for Sustainable Development.}, } @article {pmid32597701, year = {2020}, author = {Liu, Y and Guo, Z and El Smaily, M and Wang, L}, title = {A Wolbachia infection model with free boundary.}, journal = {Journal of biological dynamics}, volume = {14}, number = {1}, pages = {515-542}, doi = {10.1080/17513758.2020.1784474}, pmid = {32597701}, issn = {1751-3766}, mesh = {Animals ; Bacterial Infections/*microbiology ; Humans ; *Models, Biological ; Wolbachia/*physiology ; }, abstract = {Scientists have been seeking ways to use Wolbachia to eliminate the mosquitoes that spread human diseases. Could Wolbachia be the determining factor in controlling the mosquito-borne infectious diseases? To answer this question mathematically, we develop a reaction-diffusion model with free boundary in a one-dimensional environment. We divide the female mosquito population into two groups: one is the uninfected mosquito population that grows in the whole region while the other is the mosquito population infected with Wolbachia that occupies a finite small region. The mosquito population infected with Wolbachia invades the environment with a spreading front governed by a free boundary satisfying the well-known one-phase Stefan condition. For the resulting free boundary problem, we establish criteria under which spreading and vanishing occur. Our results provide useful insights on designing a feasible mosquito releasing strategy that infects the whole mosquito population with Wolbachia and eradicates the mosquito-borne diseases eventually.}, } @article {pmid32589643, year = {2020}, author = {Ajene, IJ and Khamis, FM and van Asch, B and Pietersen, G and Rasowo, BA and Ombura, FL and Wairimu, AW and Akutse, KS and Sétamou, M and Mohamed, S and Ekesi, S}, title = {Microbiome diversity in Diaphorina citri populations from Kenya and Tanzania shows links to China.}, journal = {PloS one}, volume = {15}, number = {6}, pages = {e0235348}, pmid = {32589643}, issn = {1932-6203}, mesh = {Animals ; *Biodiversity ; China ; Drug Resistance, Microbial/genetics ; Hemiptera/*microbiology ; Insect Vectors/microbiology ; Kenya ; *Microbiota/drug effects/genetics ; *Phylogeny ; Plant Diseases/microbiology ; Sequence Analysis ; Symbiosis ; Tanzania ; }, abstract = {The Asian citrus psyllid (Diaphorina citri) is a key pest of Citrus spp. worldwide, as it acts as a vector for "Candidatus Liberibacter asiaticus (Las)", the bacterial pathogen associated with the destructive Huanglongbing (HLB) disease. Recent detection of D. citri in Africa and reports of Las-associated HLB in Ethiopia suggest that the citrus industry on the continent is under imminent threat. Endosymbionts and gut bacteria play key roles in the biology of arthropods, especially with regards to vector-pathogen interactions and resistance to antibiotics. Thus, we aim to profile the bacterial genera and to identify antibiotic resistance genes within the microbiome of different populations worldwide of D. citri. The metagenome of D. citri was sequenced using the Oxford Nanopore full-length 16S metagenomics protocol, and the "What's in my pot" (WIMP) analysis pipeline. Microbial diversity within and between D. citri populations was assessed, and antibiotic resistance genes were identified using the WIMP-ARMA workflow. The most abundant genera were key endosymbionts of D. citri ("Candidatus Carsonella", "Candidatus Profftella", and Wolbachia). The Shannon diversity index showed that D. citri from Tanzania had the highest diversity of bacterial genera (1.92), and D. citri from China had the lowest (1.34). The Bray-Curtis dissimilarity showed that China and Kenya represented the most diverged populations, while the populations from Kenya and Tanzania were the least diverged. The WIMP-ARMA analyses generated 48 CARD genes from 13 bacterial species in each of the populations. Spectinomycin resistance genes were the most frequently found, with an average of 65.98% in all the populations. These findings add to the knowledge on the diversity of the African D. citri populations and the probable introduction source of the psyllid in these African countries.}, } @article {pmid32588179, year = {2020}, author = {Aydın, MF and Altay, K and Aytmirzakizi, A and Dumanlı, N}, title = {First Molecular Detection of Dirofilaria immitis and D. repens in Dogs from Kyrgyzstan.}, journal = {Acta parasitologica}, volume = {65}, number = {4}, pages = {949-953}, doi = {10.1007/s11686-020-00245-8}, pmid = {32588179}, issn = {1896-1851}, mesh = {Animals ; Cats ; *Dirofilaria immitis/genetics ; *Dirofilaria repens/genetics ; *Dirofilariasis/diagnosis/epidemiology ; *Dog Diseases/epidemiology ; Dogs ; Kyrgyzstan/epidemiology ; }, abstract = {BACKGROUND: Dirofilaria immitis and Dirofilaria repens are the causative agents of cardiopulmonary and subcutaneous dirofilariosis, respectively. This neglected disease mainly seen in dogs, cats and wild carnivores is re-emerging recent years. No study was conducted on dirofilariosis in dogs in Kyrgyzstan.

PURPOSE: The goal of this study was to investigate Dirofilaria species using PCR and sequencing in dogs from Kyrgyzstan.

METHOD: Dirofilaria spp. infection in dogs was screened via convential PCR and sequencing in 337 dogs from Kyrgyzstan.

RESULT: The overall prevalence of Dirofilaria spp. was 0.59% (2/337): DNA of D. immitis was detected in one sample and DNA of D. repens in second positive sample. In second sample, parallel co-infection of D. repens with Wolbachia was also found. While D. immitis sequence showed 98.70-100% similarity with previously reported sequences of D. immitis from dog blood, D. repens shared 100% identity with other sequences of D. repens.

CONCLUSION: These results provided first evidence for Dirofilaria spp. in Kyrgyzstan and emphasized the veterinary and medical importance.}, } @article {pmid32582067, year = {2020}, author = {Mateos, M and Martinez Montoya, H and Lanzavecchia, SB and Conte, C and Guillén, K and Morán-Aceves, BM and Toledo, J and Liedo, P and Asimakis, ED and Doudoumis, V and Kyritsis, GA and Papadopoulos, NT and Augustinos, AA and Segura, DF and Tsiamis, G}, title = {Wolbachia pipientis Associated With Tephritid Fruit Fly Pests: From Basic Research to Applications.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {1080}, pmid = {32582067}, issn = {1664-302X}, abstract = {Members of the true fruit flies (family Tephritidae) are among the most serious agricultural pests worldwide, whose control and management demands large and costly international efforts. The need for cost-effective and environmentally friendly integrated pest management (IPM) has led to the development and implementation of autocidal control strategies. These approaches include the widely used sterile insect technique and the incompatible insect technique (IIT). IIT relies on maternally transmitted bacteria (namely Wolbachia) to cause a conditional sterility in crosses between released mass-reared Wolbachia-infected males and wild females, which are either uninfected or infected with a different Wolbachia strain (i.e., cytoplasmic incompatibility; CI). Herein, we review the current state of knowledge on Wolbachia-tephritid interactions including infection prevalence in wild populations, phenotypic consequences, and their impact on life history traits. Numerous pest tephritid species are reported to harbor Wolbachia infections, with a subset exhibiting high prevalence. The phenotypic effects of Wolbachia have been assessed in very few tephritid species, due in part to the difficulty of manipulating Wolbachia infection (removal or transinfection). Based on recent methodological advances (high-throughput DNA sequencing) and breakthroughs concerning the mechanistic basis of CI, we suggest research avenues that could accelerate generation of necessary knowledge for the potential use of Wolbachia-based IIT in area-wide integrated pest management (AW-IPM) strategies for the population control of tephritid pests.}, } @article {pmid32581639, year = {2020}, author = {Khadka, S and Proshad, R and Thapa, A and Acharya, KP and Kormoker, T}, title = {Wolbachia: a possible weapon for controlling dengue in Nepal.}, journal = {Tropical medicine and health}, volume = {48}, number = {}, pages = {50}, pmid = {32581639}, issn = {1348-8945}, abstract = {Dengue, a mosquito-borne viral infectious disease, causes a high morbidity and mortality in tropical and subtropical areas of the world. In Nepal, the first case of dengue was reported in 2004 followed by frequent outbreaks in subsequent years, with the largest being in 2019 taking the death toll of six. It is reported that the number of dengue fever cases are soaring in Nepal spreading from the plains to more hilly regions. This might have serious public health implications in the future when combined with other factors, such as: global warming, lack of early detection and treatment of dengue, lack of diagnostic facilities, poor healthcare systems and mosquito control strategies. Nepal, thus, needs a cost-effective mosquito control strategy for the prevention and control of dengue. The Wolbachia-mediated biological method of the dengue control strategy is novel, economic, and environment-friendly. It has been successfully trialed in several areas of dengue-prone countries of the world, including Australia, Malaysia, Vietnam etc. resulting in significant reductions in dengue incidence. Given the lack of effective vector control strategy and weak economic condition of the country along with the persistence of climate and environment conditions that favors the host (Aedes mosquito) for Wolbachia, this approach can be a promising option to control dengue in Nepal.}, } @article {pmid32573833, year = {2020}, author = {Lucek, K and Butlin, RK and Patsiou, T}, title = {Secondary contact zones of closely-related Erebia butterflies overlap with narrow phenotypic and parasitic clines.}, journal = {Journal of evolutionary biology}, volume = {33}, number = {9}, pages = {1152-1163}, doi = {10.1111/jeb.13669}, pmid = {32573833}, issn = {1420-9101}, mesh = {Animals ; Butterflies/anatomy & histology/*genetics/microbiology ; Ecosystem ; *Gene Flow ; Hybridization, Genetic ; Phenotype ; *Reproductive Isolation ; Switzerland ; Wings, Animal/anatomy & histology ; Wolbachia/genetics ; }, abstract = {Zones of secondary contact between closely related taxa are a common legacy of the Quaternary ice ages. Despite their abundance, the factors that keep species apart and prevent hybridization are often unknown. Here, we study a very narrow contact zone between three closely related butterfly species of the Erebia tyndarus species complex. Using genomic data, we first determined whether gene flow occurs and then assessed whether it might be hampered by differences in chromosome number between some species. We found interspecific gene flow between sibling species that differ in karyotype by one chromosome. Conversely, only F1 hybrids occurred between two species that have the same karyotype, forming a steep genomic cline. In a second step, we fitted clines to phenotypic, ecological and parasitic data to identify the factors associated with the genetic cline. We found clines for phenotypic data and the prevalence of the endosymbiont parasite Wolbachia to overlap with the genetic cline, suggesting that they might be drivers for separating the two species. Overall, our results highlight that some gene flow is possible between closely related species despite different chromosome numbers, but that other barriers restrict such gene flow.}, } @article {pmid32571804, year = {2020}, author = {Wang, X and Kelkar, YD and Xiong, X and Martinson, EO and Lynch, J and Zhang, C and Werren, JH and Wang, X}, title = {Genome Report: Whole Genome Sequence and Annotation of the Parasitoid Jewel Wasp Nasonia giraulti Laboratory Strain RV2X[u].}, journal = {G3 (Bethesda, Md.)}, volume = {10}, number = {8}, pages = {2565-2572}, pmid = {32571804}, issn = {2160-1836}, mesh = {Animals ; Genome ; Genomics ; Laboratories ; *Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {Jewel wasps in the genus of Nasonia are parasitoids with haplodiploidy sex determination, rapid development and are easy to culture in the laboratory. They are excellent models for insect genetics, genomics, epigenetics, development, and evolution. Nasonia vitripennis (Nv) and N. giraulti (Ng) are closely-related species that can be intercrossed, particularly after removal of the intracellular bacterium Wolbachia, which serve as a powerful tool to map and positionally clone morphological, behavioral, expression and methylation phenotypes. The Nv reference genome was assembled using Sanger, PacBio and Nanopore approaches and annotated with extensive RNA-seq data. In contrast, Ng genome is only available through low coverage resequencing. Therefore, de novo Ng assembly is in urgent need to advance this system. In this study, we report a high-quality Ng assembly using 10X Genomics linked-reads with 670X sequencing depth. The current assembly has a genome size of 259,040,977 bp in 3,160 scaffolds with 38.05% G-C and a 98.6% BUSCO completeness score. 97% of the RNA reads are perfectly aligned to the genome, indicating high quality in contiguity and completeness. A total of 14,777 genes are annotated in the Ng genome, and 72% of the annotated genes have a one-to-one ortholog in the Nv genome. We reported 5 million Ng-Nv SNPs which will facility mapping and population genomic studies in Nasonia In addition, 42 Ng-specific genes were identified by comparing with Nv genome and annotation. This is the first de novo assembly for this important species in the Nasonia model system, providing a useful new genomic toolkit.}, } @article {pmid32571427, year = {2020}, author = {Laidoudi, Y and Davoust, B and Varloud, M and Niang, EHA and Fenollar, F and Mediannikov, O}, title = {Development of a multiplex qPCR-based approach for the diagnosis of Dirofilaria immitis, D. repens and Acanthocheilonema reconditum.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {319}, pmid = {32571427}, issn = {1756-3305}, mesh = {Angiostrongylus/genetics/isolation & purification/microbiology ; Animals ; Antigens, Helminth/blood ; Bacterial Proteins/genetics ; Diagnosis, Differential ; Dog Diseases/*diagnosis/parasitology ; Dogs ; Filariasis/diagnosis/parasitology/*veterinary ; Filarioidea/*classification/genetics/*isolation & purification/microbiology ; Genotype ; Helminth Proteins/genetics ; Molecular Diagnostic Techniques ; Multilocus Sequence Typing ; Real-Time Polymerase Chain Reaction ; Sensitivity and Specificity ; Wolbachia/genetics/isolation & purification ; }, abstract = {BACKGROUND: Dirofilaria immitis, D. repens and Acanthocheilonema reconditum are the main causative agents of zoonotic canine filariosis.

METHODS: We developed a combined multiplex approach for filaria and Wolbachia detection using the 28S-based pan-filarial and 16S-based pan-Wolbachia qPCRs, respectively, involving a fast typing method of positive samples using triplex qPCR targeting A. reconditum, D. immitis and D. repens, and a duplex qPCR targeting Wolbachia of D. immitis and D. repens. The approach was complemented by a duplex qPCR for the differential diagnosis of heartworms (D. immitis and Angiostrongylus vasorum) and pan-filarial cox1 and pan-Wolbachia ftsZ PCRs to identify other filarial parasites and their Wolbachia, respectively. A total of 168 canine blood and sera samples were used to validate the approach. Spearman's correlation was used to assess the association between filarial species and the strain of Wolbachia. Positive samples for both the heartworm antigen-test after heating sera and at least one DNA-positive for D. immitis and its Wolbachia were considered true positive for heartworm infection. Indeed, the presence of D. repens DNA or that of its Wolbachia as well as A. reconditum DNA indicates true positive infections.

RESULTS: The detection limit for Wolbachia and filariae qPCRs ranged from 5 × 10[-1] to 1.5 × 10[-4] mf/ml of blood. When tested on clinical samples, 29.2% (49/168) tested positive for filariae or Wolbachia DNA. Filarial species and Wolbachia genotypes were identified by the combined multiplex approach from all positive samples. Each species of Dirofilaria was significantly associated with a specific genotype of Wolbachia. Compared to the true positives, the approach showed excellent agreement (k = 0.98-1). Unlike D. immitis DNA, no A. vasorum DNA was detected by the duplex qPCR. The immunochromatographic test for heartworm antigen showed a substantial (k = 0.6) and a weak (k = 0.15) agreements before and after thermal pre-treatment of sera, respectively.

CONCLUSIONS: The proposed approach is a reliable tool for the exploration and diagnosis of occult and non-occult canine filariosis. The current diagnosis of heartworm disease based on antigen detection should always be confirmed by qPCR essays. Sera heat pre-treatment is not effective and strongly discouraged.}, } @article {pmid32560686, year = {2020}, author = {Driscoll, TP and Verhoeve, VI and Gillespie, JJ and Johnston, JS and Guillotte, ML and Rennoll-Bankert, KE and Rahman, MS and Hagen, D and Elsik, CG and Macaluso, KR and Azad, AF}, title = {A chromosome-level assembly of the cat flea genome uncovers rampant gene duplication and genome size plasticity.}, journal = {BMC biology}, volume = {18}, number = {1}, pages = {70}, pmid = {32560686}, issn = {1741-7007}, support = {R01 AI126853/AI/NIAID NIH HHS/United States ; R01 AI017828/AI/NIAID NIH HHS/United States ; R21 AI146773/AI/NIAID NIH HHS/United States ; R01 AI122672/AI/NIAID NIH HHS/United States ; R21AI26108/NH/NIH HHS/United States ; T32 AI095190/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Chromosomes ; Ctenocephalides/*genetics ; *DNA Copy Number Variations ; Female ; *Gene Duplication ; *Genome Size ; Male ; }, abstract = {BACKGROUND: Fleas (Insecta: Siphonaptera) are small flightless parasites of birds and mammals; their blood-feeding can transmit many serious pathogens (i.e., the etiological agents of bubonic plague, endemic and murine typhus). The lack of flea genome assemblies has hindered research, especially comparisons to other disease vectors. Accordingly, we sequenced the genome of the cat flea, Ctenocephalides felis, an insect with substantial human health and veterinary importance across the globe.

RESULTS: By combining Illumina and PacBio sequencing of DNA derived from multiple inbred female fleas with Hi-C scaffolding techniques, we generated a chromosome-level genome assembly for C. felis. Unexpectedly, our assembly revealed extensive gene duplication across the entire genome, exemplified by ~ 38% of protein-coding genes with two or more copies and over 4000 tRNA genes. A broad range of genome size determinations (433-551 Mb) for individual fleas sampled across different populations supports the widespread presence of fluctuating copy number variation (CNV) in C. felis. Similarly, broad genome sizes were also calculated for individuals of Xenopsylla cheopis (Oriental rat flea), indicating that this remarkable "genome-in-flux" phenomenon could be a siphonapteran-wide trait. Finally, from the C. felis sequence reads, we also generated closed genomes for two novel strains of Wolbachia, one parasitic and one symbiotic, found to co-infect individual fleas.

CONCLUSION: Rampant CNV in C. felis has dire implications for gene-targeting pest control measures and stands to complicate standard normalization procedures utilized in comparative transcriptomics analysis. Coupled with co-infection by novel Wolbachia endosymbionts-potential tools for blocking pathogen transmission-these oddities highlight a unique and underappreciated disease vector.}, } @article {pmid32559297, year = {2020}, author = {Wangkeeree, J and Tewaruxsa, P and Roddee, J and Hanboonsong, Y}, title = {Wolbachia (Rickettsiales: Alphaproteobacteria) Infection in the Leafhopper Vector of Sugarcane White Leaf Disease.}, journal = {Journal of insect science (Online)}, volume = {20}, number = {3}, pages = {}, pmid = {32559297}, issn = {1536-2442}, mesh = {Animals ; Female ; Hemiptera/growth & development/*microbiology ; Insect Vectors/growth & development/microbiology ; Male ; Nymph/growth & development/microbiology ; Ovum/growth & development/microbiology ; Plant Diseases/microbiology ; Saccharum/microbiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a maternally inherited bacterium ubiquitous in insects that has attracted interest as a prospective insect pest-control agent. Here, we detected and characterized Wolbachia in the leafhoppers Matsumuratettix hiroglyphicus (Matsumura) (Cicadellidae: Hemiptera) and Yamatotettix flavovittatus Matsumura (Cicadellidae: Hemiptera), insect vectors of the phytoplasma that cause white leaf disease in sugarcane. The 16S rRNA and wsp gene markers revealed that Wolbachia was not present in the M. hiroglyphicus but naturally occurs in Y. flavovittatus. Additionally, the infection rates in adult leafhoppers ranged from 0 to 100% depending on geographic location. Moreover, Wolbachia was detected in the eggs and first- to fifth-instar nymphs of Y. flavovittatus. A phylogenic tree of Wolbachia indicated that it resided in the monophyletic supergroup B clade and clustered in the Ori subgroup. Furthermore, fluorescence in situ hybridization revealed that Wolbachia localized to the egg apices, randomly distributed in the egg cytoplasm, and was concentrated in the nymph and adult bacteriomes, as well as occasional detection in the thorax and abdomen. To the best of our knowledge, the present study is the first to demonstrate the prevalence of Wolbachia in the leafhopper Y. flavovittatus. The obtained results would provide useful information for the future development of Wolbachia as a biological control agent for the leafhopper vectors.}, } @article {pmid34392954, year = {2020}, author = {Ciuca, L and Vismarra, A and Lebon, W and Beugnet, F and Morchon, R and Rinaldi, L and Cringoli, G and Kramer, L and Genchi, M}, title = {New insights into the biology, diagnosis and immune response to Dirofilaria repens in the canine host.}, journal = {Veterinary parasitology}, volume = {277S}, number = {}, pages = {100029}, doi = {10.1016/j.vpoa.2020.100029}, pmid = {34392954}, issn = {1873-2550}, abstract = {Dogs are the primary host for Dirofilaria repens, therefore it is mandatory to accurately diagnose the canine infection and to expand our current knowledge on parasite biology and the immune response of the infected host for a better prevention.Thus, the aim of the present study was to provide new insights from experimental infections of dogs with D. repens, focusing on the evaluation of: 1) the pre-patent period and 2) the antibody response against D. repens somatic antigens and against the Wolbachia endosymbiont. Briefly, on Day 0, twenty purpose-bred Beagle dogs were experimentally infected with 50 infective larvae (L3) of D. repens. Starting from Day 58 until the last day of the study (Day 281), blood samples were collected on a monthly basis for detection of antibodies against D. repens (Dr) and recombinant Wolbachia surface protein (rWSP) by non-commercial IgG-ELISAs. Additional samples were collected on Days 220, 245 and 281 for the detection of microfilariae (mff) using the modified Knott's test and biomolecular analysis, following two PCR protocols: Gioia et al. (2010; protocol A) and Rishniw et al. (2006- protocol B). The results were analysed by univariate statistical analyses using 2×2 contingency tables and K Cohen was calculated to assess the agreement among all the diagnostic techniques. Overall, the outcome of the study revealed that out of the 20 dogs experimentally infected with D. repens, 16 (80 %) were microfilaraemic, 17 (85 %) were positive at DNA detection in the blood, 18 (90 %) had D. repens antibodies and 16 (80 %) had Wolbachia antibodies on the last day of the study. The overall k agreement between Knott's and PCR protocol B was 0.442 (P=0.0001) and increased throughout the study, reaching 0.828 (P=0.0001) on Day 281. To the authors knowledge, this is only the second study reporting antibody response to D. repens somatic antigen in experimentally infected dogs. ELISA results showed that an antibody response develops before the onset of patency, and steadily increases with time. Results would suggest that the development of an immunological response to infection could lead to application in epidemiological studies, risk assessment and as an aid in the diagnostic approach in dogs, in particular for early infections without mff.}, } @article {pmid34900237, year = {2019}, author = {Velez, ID and Santacruz, E and Kutcher, SC and Duque, SL and Uribe, A and Barajas, J and Gonzalez, S and Patino, AC and Zuluaga, L and Martínez, L and Muñoz, E and Mejia, MC and Arbelaez, MP and Pulido, H and Jewell, NP and Dufault, SM and O'Neill, SL and Simmons, CP and Anders, KL and Tanamas, SK}, title = {The impact of city-wide deployment of Wolbachia-carrying mosquitoes on arboviral disease incidence in Medellín and Bello, Colombia: study protocol for an interrupted time-series analysis and a test-negative design study.}, journal = {F1000Research}, volume = {8}, number = {}, pages = {1327}, doi = {10.12688/f1000research.19858.2}, pmid = {34900237}, issn = {2046-1402}, abstract = {Background: Dengue, chikungunya and Zika are viral infections transmitted by Aedes aegypti mosquitoes, and present major public health challenges in tropical regions. Traditional vector control methods have been ineffective at halting disease transmission. The World Mosquito Program has developed a novel approach to arbovirus control using Ae. aegypti stably transfected with the Wolbachia bacterium, which have significantly reduced ability to transmit dengue, Zika and chikungunya in laboratory experiments. Field releases in eight countries have demonstrated Wolbachia establishment in local Ae. aegypti populations. Methods: We describe a pragmatic approach to measuring the epidemiological impact of city-wide Wolbachia deployments in Bello and Medellín, Colombia. First, an interrupted time-series analysis will compare the incidence of dengue, chikungunya and Zika case notifications before and after Wolbachia releases, across the two municipalities. Second, a prospective case-control study using a test-negative design will be conducted in one quadrant of Medellín. Three of the six contiguous release zones in the case-control area were allocated to receive the first Wolbachia deployments in the city and three to be treated last, approximating a parallel two-arm trial for the >12-month period during which Wolbachia exposure remains discordant. Allocation, although non-random, aimed to maximise balance between arms in historical dengue incidence and demographics. Arboviral disease cases and arbovirus-negative controls will be enrolled concurrently from febrile patients presenting to primary care, with case/control status classified retrospectively following laboratory diagnostic testing. Intervention effect is estimated from an aggregate odds ratio comparing Wolbachia-exposure odds among test-positive cases versus test-negative controls. Discussion: The study findings will add to an accumulating body of evidence from global field sites on the efficacy of the Wolbachia method in reducing arboviral disease incidence, and can inform decisions on wider public health implementation of this intervention in the Americas and beyond. Trial registration: ClinicalTrials.gov: NCT03631719. Registered on 15 August 2018.}, } @article {pmid33447371, year = {2019}, author = {Durovni, B and Saraceni, V and Eppinghaus, A and Riback, TIS and Moreira, LA and Jewell, NP and Dufault, SM and O'Neill, SL and Simmons, CP and Tanamas, SK and Anders, KL}, title = {The impact of large-scale deployment of Wolbachia mosquitoes on dengue and other Aedes-borne diseases in Rio de Janeiro and Niterói, Brazil: study protocol for a controlled interrupted time series analysis using routine disease surveillance data.}, journal = {F1000Research}, volume = {8}, number = {}, pages = {1328}, pmid = {33447371}, issn = {2046-1402}, mesh = {Aedes/*virology ; Animals ; Biological Control Agents ; Brazil/epidemiology ; *Chikungunya Fever/epidemiology/prevention & control ; Cities ; *Dengue/epidemiology/prevention & control ; Dengue Virus ; Humans ; Incidence ; Interrupted Time Series Analysis ; *Mosquito Control ; Mosquito Vectors/virology ; *Wolbachia ; Zika Virus ; *Zika Virus Infection/epidemiology/prevention & control ; }, abstract = {Background: Rio de Janeiro and Niterói are neighbouring cities in southeastern Brazil which experience large dengue epidemics every 2 to 5 years, with >100,000 cases notified in epidemic years. Costs of vector control and direct and indirect costs due to the Aedes-borne diseases dengue, chikungunya and Zika were estimated to total $650 million USD in 2016, but traditional vector control strategies have not been effective in preventing mosquito-borne disease outbreaks. The Wolbachia method is a novel and self-sustaining approach for the biological control of Aedes-borne diseases, in which the transmission potential of Aedes aegypti mosquitoes is reduced by stably transfecting them with the Wolbachia bacterium (wMel strain). This paper describes a study protocol for evaluating the effect of large-scale non-randomised releases of Wolbachia--infected mosquitoes on the incidence of dengue, Zika and chikungunya in the two cities of Niterói and Rio de Janeiro. This follows a lead-in period since 2014 involving intensive community engagement, regulatory and public approval, entomological surveys, and small-scale pilot releases. Method: The Wolbachia releases during 2017-2019 covered a combined area of 170 km [2] with a resident population of 1.2 million, across Niterói and Rio de Janeiro. Untreated areas with comparable historical dengue profiles and demographic characteristics have been identified a priori as comparative control areas in each city. The proposed pragmatic epidemiological approach combines a controlled interrupted time series analysis of routinely notified suspected and laboratory-confirmed dengue and chikungunya cases, together with monitoring of Aedes-borne disease activity utilising outbreak signals routinely used in public health disease surveillance. Discussion: If the current project is successful, this model for control of mosquito-borne disease through Wolbachia releases can be expanded nationally and regionally.}, } @article {pmid33601807, year = {2018}, author = {Noor-Shazleen-Husnie, MM and Emelia, O and Ahmad-Firdaus, MS and Zainol-Ariffin, P and Aishah-Hani, A}, title = {Detection of Wolbachia in wild mosquito populations from selected areas in Peninsular Malaysia by loop-mediated isothermal amplification (LAMP) technique.}, journal = {Tropical biomedicine}, volume = {35}, number = {2}, pages = {330-346}, pmid = {33601807}, issn = {2521-9855}, abstract = {Wolbachia, a naturally endosymbiotic bacteria, has shown its potential as one of biological control agents for vector borne diseases. Due to large number of mosquitoes involved in Wolbachia screening, Loop-mediated isothermal amplification (LAMP) is recommended as a convenient and time-saving technique. This study aimed to evaluate a newly developed LAMP assay for detection of Wolbachia by targeting 16S rDNA gene in samples of wild mosquito populations. The LAMP products were confirmed by colorimetric detection using hydroxy naphthol blue (HNB), digestion with RsaI restriction enzyme and gel electrophoresis. The restriction enzyme digestion of PCR products was performed to differentiate between Wolbachia supergroups A and B. Out of 765 mosquito samples tested, 349 (45.6%) and 237 (31%) of the samples were positive for LAMP and PCR techniques respectively. The prevalence of Wolbachia detected in mosquitoes was significantly higher using LAMP as compared to PCR. There is significant association between numbers of mosquitoes positive with Wolbachia detected using LAMP and PCR (χ[2]=61.31; df=1; p < 0.05) with a kappa (κ) value of 0.27 and Phi value, 0.283. This study highlighted the potential of LAMP as a sensitive, specific and rapid tool for screening of Wolbachia in mosquitoes, thus it presents as an alternative to PCR-based assays.}, } @article {pmid33592944, year = {2017}, author = {Vythilingam, I and Wan-Yusoff, WS}, title = {Dengue vector control in Malaysia: Are we moving in the right direction?.}, journal = {Tropical biomedicine}, volume = {34}, number = {4}, pages = {746-758}, pmid = {33592944}, issn = {2521-9855}, abstract = {Dengue is a major public health problem across more than 123 countries. Vector control has been the hallmark of the dengue control programme in many countries in Southeast Asia since there are no anti-dengue drugs available, and the most recent dengue vaccine is partly efficacious. House-to-house larval surveys, source reduction, larviciding, fogging, ULV which have been carried out since the inception of the dengue control programme in the 1970s are no longer practicable and need to be augmented by more targeted but less ambitious outbreak responses that focus on a few tools that might justify expense of deployment. However, according to recent reports these tools have not really been evaluated for their effectiveness in dengue control. Novel techniques such the release of genetically modified mosquitoes (RIDL) and the use of the bacterium Wolbachia to control the populations of the Ae. aegypti are still under trial. In this review proactive methods to detect epidemics have been suggested. Tools based on adult mosquitoes is an important strategy for dengue vector surveillance and control. The outbreak response may be more efficient when timely vector control measures are implemented after the immediate detection of an infected mosquito.}, } @article {pmid34541196, year = {2017}, author = {Fink, C and Von Frieling, J and Knop, M and Roeder, T}, title = {Drosophila Fecal Sampling.}, journal = {Bio-protocol}, volume = {7}, number = {18}, pages = {e2547}, pmid = {34541196}, issn = {2331-8325}, abstract = {Fecal sampling is a non-invasive method which raises the possibility to study the development and the changes in the microbial community throughout different time points of a fly population or throughout different treatments. This method allows precise manipulation to trigger the fly's physiology by nutritional interventions, bacterial infections or other stressors. As in most other animals, the intestinal microbiota is essential for a healthy fly-life. Because Drosophila only harbors a relative simple bacterial community with a small variety of round about 8 to 10 different species, it is rather easy to build up the microbial community and to investigate microbial changes after treatment. Another positive effect using the fly's feces is that bacteria that are not part of the intestinal microbiome, for example Wolbachia, can be excluded directly from the analysis because they are not excreted. Using this method, the generated datasets may reflect a good paradigm to study microbiome associated diseases in a simple fly model or furthermore, to test drugs in a high-throughput approach.}, } @article {pmid33592927, year = {2017}, author = {Teo, CHJ and Lim, PKC and Voon, K and Mak, JW}, title = {Detection of dengue viruses and Wolbachia in Aedes aegypti and Aedes albopictus larvae from four urban localities in Kuala Lumpur, Malaysia.}, journal = {Tropical biomedicine}, volume = {34}, number = {3}, pages = {583-597}, pmid = {33592927}, issn = {2521-9855}, abstract = {Dengue fever (DF) is currently one of the most important mosquito-borne diseases that affects humans. Dengue fever (DF) and dengue hemorrhagic fever (DHF) are caused by four serotypes of dengue viruses (DENV-1 to DENV-4). The main vector transmitting dengue is Aedes aegypti while Aedes albopictus acts as a secondary vector. As treatment is unavailable and the first dengue vaccine approved in Mexico, Dengvaxia® has yet to be accepted worldwide, prevention of the disease relies heavily on surveillance and control of mosquito vectors. A transgene driver, Wolbachia was found to limit the transmission of dengue virus in Aedes mosquitoes. Wolbachia alone was able to inhibit viral replication, dissemination and transmission in A. aeygpti mosquitoes in experimental studies. In A. albopictus, Wolbachia did not affect the replication of dengue virus but was able to reduce the viral infection of mosquito salivary glands and limit transmission. Studies on Wolbachia have all been carried out in adult Aedes mosquitoes, hence this study was conducted to determine the presence of dengue virus serotypes and Wolbachia in A. aegypti and A. albopictus larvae collected from ovitraps in four localities in Kuala Lumpur viz. Happy Gardens, IMU Bukit Jalil, Ampang and Taman Yarl. Another objective of this study was to determine the association between dengue virus serotypes and the presence of Wolbachia in A. aegypti and A. albopictus larvae. A total of 300 mosquito larvae was collected; 99 (Happy Gardens), 85 (Bukit Jalil), 73 (Ampang) and 43 (Taman Yarl). Out of 300 larvae collected, 284 were identified as A. albopictus and 16 others were identified as A. aegypti. Of the 284 A. albopictus larvae collected, 211 (74.3%) and 73 (25.7%) were found to be negative and positive for dengue virus respectively. The dengue serotypes detected were 2 DENV-2 (2.7%), 58 DENV-3 (79.5%) and 13 DENV-4 (17.8%). DENV-1 was not detected in any of the A. albopictus larvae. For A. aegypti, out of 16 A. aegypti larvae collected, 12 (75%) were found to be negative and 4 (25%) were positive for DENV-2. For the detection of Wolbachia in A. albopictus, 71 out of 284 (25%) and 213 (75%) larvae were found to be positive and negative for Wolbachia respectively. For A. aegypti, 4 (25%) and 12 (75%) out of 16 larvae were positive and negative for Wolbachia respectively. This is the first report of Wolbachia in A. albopictus and A. aegypti larvae in Malaysia. A chisquare test analysis to determine the association between dengue virus and Wolbachia in A. albopictus and A. aegypti larvae collected from the four localities in Kuala Lumpur showed that there was no association (χ[2] = 3.080; df = 1; P > 0.05).}, } @article {pmid32846693, year = {2015}, author = {Johnson, KN}, title = {Bacteria and antiviral immunity in insects.}, journal = {Current opinion in insect science}, volume = {8}, number = {}, pages = {97-103}, doi = {10.1016/j.cois.2015.01.008}, pmid = {32846693}, issn = {2214-5753}, abstract = {Tripartite interactions between viruses, bacteria and hosts can have significant implications for the outcome of infections. In insects recent studies have shown that these interactions can greatly influence the transmission of vector borne disease, including transmission of dengue virus. Mosquito gut bacteria modulate arbovirus infection of the mosquito host and specific bacterial isolates have been identified that negatively impact the outcome of infection. In addition, an endosymbiotic bacterium Wolbachia, which is very common in insects, protects flies and mosquitoes from virus infection. This antiviral protection has implications for both naturally infected insects and for use in disease control. This review summarises the current state of knowledge for each of these interactions with an emphasis on natural interactions.}, } @article {pmid32669893, year = {2014}, author = {Higazi, TB and Geary, TG and Mackenzie, CD}, title = {Chemotherapy in the treatment, control, and elimination of human onchocerciasis.}, journal = {Research and reports in tropical medicine}, volume = {5}, number = {}, pages = {77-93}, pmid = {32669893}, issn = {1179-7282}, abstract = {Onchocerciasis treatment is one of the most positive stories in tropical medicine although major challenges remain to reaching the ultimate goal of disease elimination. Such challenges are to be expected when the therapeutic goal is to kill and safely remove a large multistage, efficient, metazoan infectious agent such as Onchocerca volvulus that has an exceptionally complicated relationship with its host. Successful control of onchocerciasis has often been hampered by host reactions following chemotherapy, that can sometimes cause significant tissue pathology. Presence of other filariae, particularly Loa loa, in endemic onchocerciasis-treatment areas also poses severe problems due to adverse reactions caused by drug-induced death of the coincident microfilariae of this usually clinically benign species. Although ivermectin has been very successful, there is a need to enhance the progress toward elimination of onchocerciasis; new drugs and their efficient use are keys to this. The permanent absence of Onchocerca microfilaridermia, defined as the lack of resurgence of skin microfilarial loads after treatment, is the ultimate characteristic of a useful new chemotherapeutic agent. Several drugs are under investigation to achieve this, including the reassessment of currently available and previously tested agents, such as the antibiotic, doxycycline, which targets the adult parasites through its anti-Wolbachia endosymbiont activity. Flubendazole, a benzimidazole derivative approved for treatment of human gastrointestinal nematodes, is also being considered for repurposing as a macrofilaricide to aid in the achievement of eradication. The managerial challenges existing at the population level also need to be addressed; these include drug-distribution fatigue, the need to include noncompliant people, civil unrest in endemic areas, political cross-border issues, restrictions of age and pregnancy, and complications due to integration with other treatment programs. It is likely that a panel of chemotherapeutic options, new and old, supported by strong and effective distribution systems will be the best way to address challenges of treatment and elimination of this infection. Future research should also address management of treatment and control, and consider how new treatment paradigms can be incorporated to meet time lines set for global elimination by 2025.}, } @article {pmid32555677, year = {2020}, author = {Bhattacharya, T and Newton, ILG and Hardy, RW}, title = {Viral RNA is a target for Wolbachia-mediated pathogen blocking.}, journal = {PLoS pathogens}, volume = {16}, number = {6}, pages = {e1008513}, pmid = {32555677}, issn = {1553-7374}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; R21 AI121849/AI/NIAID NIH HHS/United States ; R21 AI153785/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes ; Animals ; Cell Line ; Chlorocebus aethiops ; Cricetinae ; Drosophila melanogaster ; Flavivirus/genetics/*metabolism ; RNA, Viral/genetics/*metabolism ; Togaviridae/genetics/*metabolism ; Vero Cells ; Wolbachia/genetics/*metabolism ; }, abstract = {The ability of the endosymbiont Wolbachia pipientis to restrict RNA viruses is presently being leveraged to curb global transmission of arbovirus-induced diseases. Past studies have shown that virus replication is limited early in arthropod cells colonized by the bacterium, although it is unclear if this phenomenon is replicated in mosquito cells that first encounter viruses obtained through a vertebrate blood meal. Furthermore, these cellular events neither explain how Wolbachia limits dissemination of viruses between mosquito tissues, nor how it prevents transmission of infectious viruses from mosquitoes to vertebrate host. In this study, we try to address these issues using an array of mosquito cell culture models, with an additional goal being to identify a common viral target for pathogen blocking. Our results establish the viral RNA as a cellular target for Wolbachia-mediated inhibition, with the incoming viral RNA experiencing rapid turnover following internalization in cells. This early block in replication in mosquito cells initially infected by the virus thus consequently reduces the production of progeny viruses from these same cells. However, this is not the only contributor to pathogen blocking. We show that the presence of Wolbachia reduces the per-particle infectivity of progeny viruses on naïve mosquito and vertebrate cells, consequently limiting virus dissemination and transmission, respectively. Importantly, we demonstrate that this aspect of pathogen blocking is independent of any particular Wolbachia-host association and affects viruses belonging to Togaviridae and Flaviviridae families of RNA viruses. Finally, consistent with the idea of the viral RNA as a target, we find that the encapsidated virion RNA is less infectious for viruses produced from Wolbachia-colonized cells. Collectively, our findings present a common mechanism of pathogen blocking in mosquitoes that establish a link between virus inhibition in the cell to virus dissemination and transmission.}, } @article {pmid32546497, year = {2020}, author = {Hague, MTJ and Mavengere, H and Matute, DR and Cooper, BS}, title = {Environmental and Genetic Contributions to Imperfect wMel-Like Wolbachia Transmission and Frequency Variation.}, journal = {Genetics}, volume = {215}, number = {4}, pages = {1117-1132}, pmid = {32546497}, issn = {1943-2631}, support = {R01 GM121750/GM/NIGMS NIH HHS/United States ; R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/classification/genetics/growth & development/*microbiology ; Environmental Exposure/*analysis ; Female ; *Gene-Environment Interaction ; Gram-Negative Bacterial Infections/microbiology/*transmission ; *Host-Pathogen Interactions ; Male ; Wolbachia/*pathogenicity ; }, abstract = {Maternally transmitted Wolbachia bacteria infect about half of all insect species. They usually show imperfect maternal transmission and often produce cytoplasmic incompatibility (CI). Irrespective of CI, Wolbachia frequencies tend to increase when rare only if they benefit host fitness. Several Wolbachia, including wMel that infects Drosophila melanogaster, cause weak or no CI and persist at intermediate frequencies. On the island of São Tomé off West Africa, the frequencies of wMel-like Wolbachia infecting Drosophila yakuba (wYak) and Drosophila santomea (wSan) fluctuate, and the contributions of imperfect maternal transmission, fitness effects, and CI to these fluctuations are unknown. We demonstrate spatial variation in wYak frequency and transmission on São Tomé. Concurrent field estimates of imperfect maternal transmission do not predict spatial variation in wYak frequencies, which are highest at high altitudes where maternal transmission is the most imperfect. Genomic and genetic analyses provide little support for D. yakuba effects on wYak transmission. Instead, rearing at cool temperatures reduces wYak titer and increases imperfect transmission to levels observed on São Tomé. Using mathematical models of Wolbachia frequency dynamics and equilibria, we infer that temporally variable imperfect transmission or spatially variable effects on host fitness and reproduction are required to explain wYak frequencies. In contrast, spatially stable wSan frequencies are plausibly explained by imperfect transmission, modest fitness effects, and weak CI. Our results provide insight into causes of wMel-like frequency variation in divergent hosts. Understanding this variation is crucial to explain Wolbachia spread and to improve wMel biocontrol of human disease in transinfected mosquito systems.}, } @article {pmid32545688, year = {2020}, author = {Suwantika, AA and Kautsar, AP and Supadmi, W and Zakiyah, N and Abdulah, R and Ali, M and Postma, MJ}, title = {Cost-Effectiveness of Dengue Vaccination in Indonesia: Considering Integrated Programs with Wolbachia-Infected Mosquitos and Health Education.}, journal = {International journal of environmental research and public health}, volume = {17}, number = {12}, pages = {}, pmid = {32545688}, issn = {1660-4601}, mesh = {Animals ; Child ; Cost-Benefit Analysis ; *Culicidae ; *Dengue ; Health Education ; Humans ; Indonesia ; Vaccination ; *Wolbachia ; }, abstract = {Despite the fact that morbidity and mortality rates due to dengue infection in Indonesia are relatively high, a dengue vaccination has not yet been introduced. Next to vaccination, Wolbachia-infected mosquitoes and health education have been considered to be potential interventions to prevent dengue infection in Indonesia. This study was aimed to analyse the cost-effectiveness of dengue vaccination in Indonesia whilst taking Wolbachia and health education programs into account. An age-structured decision tree model was developed to assess the cost-effectiveness. Approximately 4,701,100 children were followed-up in a 10-year time horizon within a 1-year analytical cycle. We compared three vaccination strategies: one focussing on vaccination only, another combining vaccination and a Wolbachia program, and a third scenario combining vaccination and health education. All scenarios were compared with a no-intervention strategy. The result showed that only vaccination would reduce dengue fever (DF), dengue haemorrhagic fever (DHF), and dengue shock syndrome (DSS) by 123,203; 97,140 and 283 cases, respectively. It would save treatment cost at $10.3 million and $6.2 million from the healthcare and payer perspectives, respectively. The combination of vaccination and a Wolbachia program would reduce DF, DHF and DSS by 292,488; 230,541; and 672 cases, respectively. It would also save treatment cost at $24.3 million and $14.6 million from the healthcare and payer perspectives, respectively. The combination of vaccination and health education would reduce DF, DHF, and DSS by 187,986; 148,220; and 432 cases, respectively. It would save treatment cost at $15.6 million and $9.4 million from the healthcare and payer perspectives, respectively. The incremental cost-effectiveness ratios (ICERs) from the healthcare perspective were estimated to be $9,995, $4,460, and $6,399 per quality-adjusted life year (QALY) gained for the respective scenarios. ICERs from the payer perspective were slightly higher. It can be concluded that vaccination combined with a Wolbachia program was confirmed to be the most cost-effective intervention. Dengue infection rate, vaccine efficacy, cost of Wolbachia program, underreporting factor for hospitalization, vaccine price and mortality rate were considered to be the most influential parameters affecting the ICERs.}, } @article {pmid32530921, year = {2020}, author = {Staunton, KM and Crawford, JE and Cornel, D and Yeeles, P and Desnoyer, M and Livni, J and Holeman, J and Mulligan, FS and Snoad, N and Ritchie, SA}, title = {Environmental influences on Aedes aegypti catches in Biogents Sentinel traps during a Californian "rear and release" program: Implications for designing surveillance programs.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {6}, pages = {e0008367}, pmid = {32530921}, issn = {1935-2735}, mesh = {Aedes/*physiology ; Animals ; California ; Ecosystem ; *Environmental Monitoring ; Female ; Housing ; Male ; Mosquito Control ; Mosquito Vectors/*physiology ; Wolbachia ; }, abstract = {As Aedes aegypti continues to expand its global distribution, the diseases it vectors (dengue, Zika, chikungunya and yellow fever) are of increasing concern. Modern efforts to control this species include "rear and release" strategies where lab-reared mosquitoes are distributed throughout the landscape to replace or suppress invasive populations. These programs require intensive surveillance efforts to monitor their success, and the Biogents Sentinel (BGS) trap is one of the most effective tools for sampling adult Ae. aegypti. BGS trap catches can be highly variable throughout landscapes, so we investigated the potential impacts of environmental factors on adult Ae. aegypti capture rates during a "rear and release" program in California to better understand the relative contributions of true variability in population density across a landscape and trap context. We recorded male and female Ae. aegypti catches from BGS traps, with and without CO2, throughout control sites where no mosquitoes were released and in treatment sites where males infected with Wolbachia were released. BGS trap catches were positively influenced by higher proportions of shade or bushes in the front yard of the premises as well as the presence of potential larval habitats such as subterranean vaults. In contrast, an increase in residential habitat within a 100 m radius of trap locations negatively influenced BGS trap catches. For male Ae. aegypti, increased visual complexity of the trap location positively influenced capture rates, and the presence of yard drains negatively affected catch rates in control sites. Lastly, for BGS traps using CO2, higher catch rates were noted from traps placed greater than one meter from walls or fences for both male and female mosquitoes. These results have important implications for surveillance programs of Ae. aegypti throughout the Californian urban environment including adult monitoring during "rear and release" programs.}, } @article {pmid32529355, year = {2020}, author = {Pina, T and Sabater-Muñoz, B and Cabedo-López, M and Cruz-Miralles, J and Jaques, JA and Hurtado-Ruiz, MA}, title = {Molecular characterization of Cardinium, Rickettsia, Spiroplasma and Wolbachia in mite species from citrus orchards.}, journal = {Experimental & applied acarology}, volume = {81}, number = {3}, pages = {335-355}, doi = {10.1007/s10493-020-00508-z}, pmid = {32529355}, issn = {1572-9702}, support = {AGL2014-55616-C3-3-R//Ministerio de Economía y Competitividad/ ; PICD//Universitat Jaume I/ ; FPI BES-2015-074570//Ministerio de Economía y Competitividad/ ; UJI-B2017-24//Universitat Jaume I/ ; }, mesh = {Animals ; *Citrus ; *Mites/microbiology ; Phylogeny ; *Rickettsia/genetics ; *Spiroplasma ; *Tetranychidae ; *Wolbachia/genetics ; }, abstract = {Tetranychidae spider mites are considered key citrus pests in some production areas, especially Tetranychus urticae Koch. Over the past decades, pesticide overuse seems to have promoted T. urticae population selection in citrus orchards. However, the microbiota has also been pointed out as a plausible explanation for population structure or plant host specialisation observed in several arthropod species. In this work, we have determined the incidence of Cardinium, Rickettsia, Spiroplasma and Wolbachia as representatives of major distorter bacteria genera in Aplonobia histricina (Berlese), Eutetranychus banksi (McGregor), Eutetranychus orientalis (Klein), Panonychus citri (McGregor), Tetranychus evansi Baker and Pritchard, Tetranychus turkestani Ugarov and Nikolskii, and T. urticae populations from Spanish citrus orchards. Only Wolbachia was detected by PCR. The multilocus alignment approach and phylogenetic inference indicated that all detected Wolbachia belong to supergroup B. The deep analysis of each 16S rDNA, ftsZ and wsp gene sequences allowed identifying several phylogenetically different Wolbachia sequences. It probably indicates the presence of several different races or strains, all of them belonging to supergroup B. The wsp sequence typing analysis unveiled the presence of the two already identified alleles (61 and 370) and allowed to contribute with five new alleles, supporting the presence of different but related B-races in the studied mite populations. The results are discussed and related to T. urticae population structure, previously observed in Spanish citrus orchards.}, } @article {pmid32522243, year = {2020}, author = {Madhav, M and Brown, G and Morgan, JAT and Asgari, S and McGraw, EA and James, P}, title = {Transinfection of buffalo flies (Haematobia irritans exigua) with Wolbachia and effect on host biology.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {296}, pmid = {32522243}, issn = {1756-3305}, mesh = {Animals ; Female ; Fertility ; Host Microbial Interactions ; Life Cycle Stages ; Longevity ; Male ; Microinjections ; Muscidae/*microbiology ; Pest Control, Biological/*methods ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Buffalo flies (Haematobia irritans exigua) (BF) and closely related horn flies (Haematobia irritans irritans) (HF) are invasive haematophagous parasites with significant economic and welfare impacts on cattle production. Wolbachia are intracellular bacteria found widely in insects and currently of much interest for use in novel strategies for the area wide control of insect pests and insect-vectored diseases. In this paper, we report the transinfection of BF towards the development of area-wide controls.

METHODS: Three stages of BF; embryos, pupae and adult female flies, were injected with different Wolbachia strains (wAlbB, wMel and wMelPop). The success of transinfection and infection dynamics was compared by real-time PCR and FISH and fitness effects were assessed in transinfected flies.

RESULTS: BF eggs were not easily injected because of their tough outer chorion and embryos were frequently damaged with less than 1% hatch rate of microinjected eggs. No Wolbachia infection was recorded in flies successfully reared from injected eggs. Adult and pupal injection resulted in higher survival rates and somatic and germinal tissue infections, with transmission to the succeeding generations on some occasions. Investigations of infection dynamics in flies from injected pupae confirmed that Wolbachia were actively multiplying in somatic tissues. Ovarian infections were confirmed with wMel and wMelPop in a number of instances, though not with wAlbB. Measurement of fitness traits indicated reduced longevity, decreased and delayed adult emergence, and reduced fecundity in Wolbachia-infected flies compared to mock-injected flies. Effects varied with the Wolbachia strain injected with most marked changes seen in the wMelPop-injected flies and least severe effects seen with wAlbB.

CONCLUSIONS: Adult and pupal injection were the most suitable methods for transinfecting BF and all three strains of Wolbachia successfully replicated in somatic tissues. The Wolbachia-induced fitness effects seen in transinfected BF suggest potential for use of the wMel or wMelPop strains in Wolbachia-based biocontrol programmes for BF.}, } @article {pmid32519810, year = {2021}, author = {Cai, T and Zhang, Y and Liu, Y and Deng, X and He, S and Li, J and Wan, H}, title = {Wolbachia enhances expression of NlCYP4CE1 in Nilaparvata lugens in response to imidacloprid stress.}, journal = {Insect science}, volume = {28}, number = {2}, pages = {355-362}, doi = {10.1111/1744-7917.12834}, pmid = {32519810}, issn = {1744-7917}, support = {31871991//National Natural Science Foundation of China/ ; 2019CFB471//Natural Science Foundation of Hubei Province/ ; 2662018JC049//Fundamental Research Funds for the Central Universities/ ; }, mesh = {Animals ; Cytochrome P450 Family 4/*genetics/metabolism ; *Gene Expression ; Hemiptera/*drug effects/genetics/growth & development/microbiology ; Insect Proteins/*genetics/metabolism ; Insecticide Resistance ; Insecticides/*pharmacology ; Neonicotinoids/*pharmacology ; Nitro Compounds/*pharmacology ; Nymph/drug effects/genetics/growth & development/microbiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The brown planthopper, Nilaparvata lugens, is one of the main insect pests of rice. The N. lugens gene NlCYP4CE1 encodes cytochrome P450 monooxygenase (P450), which is a key enzyme in the metabolism of the insecticide imidacloprid. Previous research has suggested that the expression of NlCYP4CE1 is induced by imidacloprid stress, but the effect of bacterial symbionts on its expression has not been determined. The results of this study show that exposure to subtoxic imidacloprid changed the structure of the bacterial symbiont community in N. lugens. Specifically, the total bacterial content increased, but the bacterial species diversity significantly decreased. Wolbachia accounted for the largest proportion of bacteria in N. lugens; its abundance significantly increased after subtoxic imidacloprid exposure. The transcript level of NlCYP4CE1 was significantly increased by imidacloprid, but this effect was significantly weakened after Wolbachia was cleared with tetracycline. This result suggests that Wolbachia enhances the expression of NlCYP4CE1 to promote the detoxification metabolic response to imidacloprid stress. Understanding the effect of bacterial symbionts on gene expression in the host provides a new perspective on interactions between insecticides and their target insect pests, and highlights that subtoxic imidacloprid exposure may raise the risk of insecticide resistance by altering the structure of bacterial symbiont communities.}, } @article {pmid32519634, year = {2020}, author = {Zueva, T and Morchón, R and Carretón, E and Ollauri-Ibáñez, C and Pericacho, M and Rodríguez-Barbero, A and Simón, F}, title = {Angiogenesis in cardiopulmonary dirofilariosis: does the Wolbachia surface protein have a pro- or anti-angiogenic effect?.}, journal = {Journal of helminthology}, volume = {94}, number = {}, pages = {e162}, doi = {10.1017/S0022149X20000450}, pmid = {32519634}, issn = {1475-2697}, mesh = {Angiogenesis Inducing Agents/*chemistry ; Animals ; Bacterial Proteins/*chemistry ; Cells, Cultured ; Dirofilaria immitis/microbiology ; Dirofilariasis/*complications/microbiology ; Dogs ; Endothelial Cells/microbiology ; Heart/parasitology ; Humans ; Inflammation ; Lung/cytology/parasitology ; Membrane Proteins/*chemistry ; Symbiosis ; Wolbachia/*chemistry ; }, abstract = {Cardiopulmonary dirofilariosis caused by Dirofilaria immitis produces inflammation, blood vessel obstruction and hypoxia, which are required conditions for the beginning of the process of neovascularization. Since D. immitis harbours intracellular symbiotic Wolbachia bacterium, the global understanding of the angiogenic process requires the analysis of the effect of the parasite molecules, but also that of Wolbachia. Canine primary lung microvascular endothelial cells were treated with the recombinant Wolbachia surface protein (rWSP) and the expression of angiogenic factors like Vascular Endothelial Growth Factor-A (VEGF-A), sFlt, membrane Endoglin (mEndoglin) and soluble Endoglin (sEndoglin), as well as the in vitro formation of pseudocapillaries, were measured. The analyses showed a significant increase in the expression of pro-angiogenic VEGF-A and anti-angiogenic sEndoglin, together with a significant decrease in both pro-angiogenic mEndoglin and pseudocapillary formation, compared to untreated controls. Due to the complexity of the angiogenic process and its relationship with other physiological processes like inflammation and fibrinolysis, these results might suggest that rWSP participate in various mechanisms related to each other and its effects might depend either on the balance between them or on the moment of their occurrence.}, } @article {pmid32516872, year = {2020}, author = {Torres-Monzón, JA and Casas-Martínez, M and López-Ordóñez, T}, title = {Infection of Aedes mosquitoes by native Wolbachia in urban cemeteries of Southern Mexico.}, journal = {Salud publica de Mexico}, volume = {62}, number = {4}, pages = {447-449}, doi = {10.21149/10163}, pmid = {32516872}, issn = {1606-7916}, mesh = {Aedes/*microbiology ; Animals ; *Cemeteries ; Female ; Mexico ; Mosquito Vectors/*microbiology ; Rain ; Species Specificity ; Wolbachia/*isolation & purification ; }, abstract = {OBJECTIVE: To evaluate the prevalence of Wolbachia infections in Aedes spp. field populations from cemeteries of Southern Mexico.

MATERIALS AND METHODS: Six cemeteries were selected to be sampled in the central part of the Soconusco region, Chiapas. Aedes albopictus and Ae. aegypti mosquitoes were collected during the rainy season of 2015. Females were analyzed individually by PCR to determine the presence of Wolbachia.

RESULTS: field overall prevalence of 38% was found; only Ae. albopictus mosquitoes were positive.

CONCLUSIONS: Local strains of Wolbachia were detected and have the potential to be applied as a biological method for vector control.}, } @article {pmid32506239, year = {2020}, author = {Liu, XC and Li, YR and Dong, B and Li, ZX}, title = {The Intruding Wolbachia Strain from the Moth Fails to Establish Itself in the Fruit Fly Due to Immune and Exclusion Reactions.}, journal = {Current microbiology}, volume = {77}, number = {9}, pages = {2441-2448}, doi = {10.1007/s00284-020-02067-3}, pmid = {32506239}, issn = {1432-0991}, support = {31772169//National Natural Science Foundation of China/ ; 31371940//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Drosophila melanogaster ; *Moths ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia is capable of regulating host reproduction, and thus of great significance in preventing the spread of insect-borne diseases and controlling pest insects. The fruit fly Drosophila melanogaster is an excellent model insect for understanding Wolbachia-host interactions. Here we artificially transferred the wCcep strain from the rice moth Corcyra cephalonica into D. melanogaster by microinjection. Crossing experiments indicated that wCcep could induce a high level of CI in the phylogenetically distant host D. melanogaster and imposed no negative fitness costs on host development and fecundity. Based on quantitative analysis, the titres of wCcep and the native wMel strain were negatively correlated, and wCcep could only be transmitted in the novel host for several generations (G0 to G4) after transinfection. Transcriptome sequencing indicated that the invading wCcep strain induced a significant immune- and stress-related response from the host. An association analysis between the expression of immune genes attacin-D/edin and the titre of Wolbachia by linear regression displayed a negative correlation between them. Our study suggest that the intrusion of wCcep elicited a robust immune response from the host and incurred a competitive exclusion from the native Wolbachia strain, which resulted in the failure of its establishment in D. melanogaster.}, } @article {pmid32502919, year = {2020}, author = {Carretón, E and Morchón, R and Falcón-Cordón, Y and Falcón-Cordón, S and Matos, JI and Montoya-Alonso, JA}, title = {Evaluation of different dosages of doxycycline during the adulticide treatment of heartworm (Dirofilaria immitis) in dogs.}, journal = {Veterinary parasitology}, volume = {283}, number = {}, pages = {109141}, doi = {10.1016/j.vetpar.2020.109141}, pmid = {32502919}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Antibodies, Bacterial/*blood ; Antinematodal Agents/administration & dosage ; Dirofilariasis/drug therapy/parasitology ; Dog Diseases/drug therapy/parasitology ; Dogs ; Dose-Response Relationship, Drug ; Doxycycline/*pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {The endosymbiont bacteria Wolbachia plays an important role in the pathogenesis and inflammatory immune response to heartworm (Dirofilaria immitis) infection in dogs. Doxycycline is used to reduce Wolbachia from all life stages of heartworm to avoid large releases of the bacteria during the death of the worms. However, the dose and duration currently recommended have been extrapolated from the treatment of other rickettsial infections. Therefore, the aim was to study the dynamics of Wolbachia IgG antibodies in heartworm-infected dogs under adulticide treatment using different dosages of doxycycline. Forty-nine heartworm-infected dogs were recruited. On day 0 (diagnosis), monthly ivermectin (6 μg/kg) was prescribed, as well as daily doxycycline for 30 days, at 10 mg/kg/12 h (n = 13), 5 mg/kg/12 h (n = 19), and 10 mg/kg/24 h (n = 17). Dogs underwent adulticide treatment and blood samples were collected on days 0, 30, 90, and 120. All dogs had antibodies against recombinant Wolbachia surface protein (rWSP), confirming the important role of the bacteria in heartworm. No significant differences were found in anti-rWSP response by presence/absence of microfilariae, or by parasite burden on day 0. In all treated groups, the anti-rWSP antibody response was not significantly different between days 0 and 30 but was significantly lower between days 0 and 120 (p < 0.05). The results of the present study suggest that the administration of a lower dose than currently recommended is sufficient to achieve a significant reduction of Wolbachia in dogs infected by D. immitis.}, } @article {pmid32498270, year = {2020}, author = {Bel Mokhtar, N and Maurady, A and Britel, MR and El Bouhssini, M and Batargias, C and Stathopoulou, P and Asimakis, E and Tsiamis, G}, title = {Detection of Wolbachia Infections in Natural and Laboratory Populations of the Moroccan Hessian Fly, Mayetiola destructor (Say).}, journal = {Insects}, volume = {11}, number = {6}, pages = {}, pmid = {32498270}, issn = {2075-4450}, support = {Erasmus+//European Union/ ; }, abstract = {Mayetiola destructor (Hessian fly) is a destructive pest of wheat in several parts of the world. Here, we investigated the presence of reproductive symbionts and the effect of the geographical location on the bacterial community associated to adult Hessian flies derived from four major wheat producing areas in Morocco. Using specific 16S rDNA PCR assay, Wolbachia infection was observed in 3% of the natural populations and 10% of the laboratory population. High throughput sequencing of V3-V4 region of the bacterial 16S rRNA gene revealed that the microbiota of adult Hessian flies was significantly influenced by their native regions. A total of 6 phyla, 10 classes and 79 genera were obtained from all the samples. Confirming the screening results, Wolbachia was identified as well in the natural Hessian flies. Phylogenetic analysis using the sequences obtained in this study indicated that there is one Wolbachia strain belonging to supergroup A. To our knowledge, this is the first report of Wolbachia in Hessian fly populations. The observed low abundance of Wolbachia most likely does not indicate induction of reproductive incompatibility. Yet, this infection may give a new insight into the use of Wolbachia for the fight against Hessian fly populations.}, } @article {pmid32491177, year = {2020}, author = {Adams, M and McBroome, J and Maurer, N and Pepper-Tunick, E and Saremi, NF and Green, RE and Vollmers, C and Corbett-Detig, RB}, title = {One fly-one genome: chromosome-scale genome assembly of a single outbred Drosophila melanogaster.}, journal = {Nucleic acids research}, volume = {48}, number = {13}, pages = {e75}, pmid = {32491177}, issn = {1362-4962}, support = {R35 GM128932/GM/NIGMS NIH HHS/United States ; R35 GM133569/GM/NIGMS NIH HHS/United States ; T32 HG008345/HG/NHGRI NIH HHS/United States ; }, mesh = {Animals ; Chromosomes, Bacterial/*genetics ; Chromosomes, Insect/*genetics ; Drosophila melanogaster/*genetics ; Genome, Bacterial/*genetics ; Genome, Insect/*genetics ; Genomics/methods ; Wolbachia/*genetics ; }, abstract = {A high quality genome assembly is a vital first step for the study of an organism. Recent advances in technology have made the creation of high quality chromosome scale assemblies feasible and low cost. However, the amount of input DNA needed for an assembly project can be a limiting factor for small organisms or precious samples. Here we demonstrate the feasibility of creating a chromosome scale assembly using a hybrid method for a low input sample, a single outbred Drosophila melanogaster. Our approach combines an Illumina shotgun library, Oxford nanopore long reads, and chromosome conformation capture for long range scaffolding. This single fly genome assembly has a N50 of 26 Mb, a length that encompasses entire chromosome arms, contains 95% of expected single copy orthologs, and a nearly complete assembly of this individual's Wolbachia endosymbiont. The methods described here enable the accurate and complete assembly of genomes from small, field collected organisms as well as precious clinical samples.}, } @article {pmid32489617, year = {2020}, author = {Zélé, F and Altıntaş, M and Santos, I and Cakmak, I and Magalhães, S}, title = {Population-specific effect of Wolbachia on the cost of fungal infection in spider mites.}, journal = {Ecology and evolution}, volume = {10}, number = {9}, pages = {3868-3880}, pmid = {32489617}, issn = {2045-7758}, abstract = {Many studies have revealed the ability of the endosymbiotic bacterium Wolbachia to protect its arthropod hosts against diverse pathogens. However, as Wolbachia may also increase the susceptibility of its host to infection, predicting the outcome of a particular Wolbachia-host-pathogen interaction remains elusive. Yet, understanding such interactions and their eco-evolutionary consequences is crucial for disease and pest control strategies. Moreover, how natural Wolbachia infections affect artificially introduced pathogens for biocontrol has never been studied. Tetranychus urticae spider mites are herbivorous crop pests, causing severe damage on numerous economically important crops. Due to the rapid evolution of pesticide resistance, biological control strategies using entomopathogenic fungi are being developed. However, although spider mites are infected with various Wolbachia strains worldwide, whether this endosymbiont protects them from fungi is as yet unknown. Here, we compared the survival of two populations, treated with antibiotics or naturally harboring different Wolbachia strains, after exposure to the fungal biocontrol agents Metarhizium brunneum and Beauveria bassiana. To control for potential effects of the bacterial community of spider mites, we also compared the susceptibility of two populations naturally uninfected by Wolbachia, treated with antibiotics or not. In one population, Wolbachia-infected mites had a better survival than uninfected ones in absence of fungi but not in their presence, whereas in the other population Wolbachia increased the mortality induced by B. bassiana. In one naturally Wolbachia-uninfected population, the antibiotic treatment increased the susceptibility of spider mites to M. brunneum, but it had no effect in the other treatments. These results suggest that natural Wolbachia infections may not hamper and may even improve the success of biological control using entomopathogenic fungi. However, they also draw caution on the generalization of such effects, given the complexity of within-host-pathogens interaction and the potential eco-evolutionary consequences of the use of biocontrol agents for Wolbachia-host associations.}, } @article {pmid32489610, year = {2020}, author = {Detcharoen, M and Arthofer, W and Jiggins, FM and Steiner, FM and Schlick-Steiner, BC}, title = {Wolbachia affect behavior and possibly reproductive compatibility but not thermoresistance, fecundity, and morphology in a novel transinfected host, Drosophila nigrosparsa.}, journal = {Ecology and evolution}, volume = {10}, number = {10}, pages = {4457-4470}, pmid = {32489610}, issn = {2045-7758}, abstract = {Wolbachia, intracellular endosymbionts, are estimated to infect about half of all arthropod species. These bacteria manipulate their hosts in various ways for their maximum benefits. The rising global temperature may accelerate species migration, and thus, horizontal transfer of Wolbachia may occur across species previously not in contact. We transinfected and then cured the alpine fly Drosophila nigrosparsa with Wolbachia strain wMel to study its effects on this species. We found low Wolbachia titer, possibly cytoplasmic incompatibility, and an increase in locomotion of both infected larvae and adults compared with cured ones. However, no change in fecundity, no impact on heat and cold tolerance, and no change in wing morphology were observed. Although Wolbachia increased locomotor activities in this species, we conclude that D. nigrosparsa may not benefit from the infection. Still, D. nigrosparsa can serve as a host for Wolbachia because vertical transmission is possible but may not be as high as in the native host of wMel, Drosophila melanogaster.}, } @article {pmid32489376, year = {2020}, author = {Khanmohammadi, M and Akhlaghi, L and Razmjou, E and Falak, R and Zolfaghari Emameh, R and Mokhtarian, K and Arshadi, M and Tasbihi, M and Meamar, AR}, title = {Morphological Description, Phylogenetic and Molecular Analysis of Dirofilaria immitis Isolated from Dogs in the Northwest of Iran.}, journal = {Iranian journal of parasitology}, volume = {15}, number = {1}, pages = {57-66}, pmid = {32489376}, issn = {1735-7020}, abstract = {BACKGROUND: Dirofilariasis is a globally distributed arthropod-borne parasitic disease of mainly canids and felids. We evaluated to extend the knowledge of morpho-molecular characteristics and outer ultrastructure of Dirofilaria immitis isolated from Northwest of Iran.

METHODS: Overall, 67 filarial worms including 41 females and 26 males parasites were collected from the cardiovascular system of the 43 stray dogs in Meshkinshar, Ardebil Province, Northwest of Iran in 2017, and subjected to light and scanning electron microscopy (SEM) as well as carmine alum staining for morpho-molecular and identification. Molecular methods were used for confirmation of morphological findings by sequencing of Cyto-chrome c oxidase subunit I (cox1) gene.

RESULTS: The partial DNA sequencing of cox1 gene of adult parasites showed considerable homology and close proximity to the previously isolated from Kerman and Meshkinshahr, Iran. The lowest genetic variation and the highest intra-species variability was found in D. immitis and Dirofilaria repens, respectively. No similarity was identified between D. immitis nucleotide sequence and Wolbachia species as its endosymbiont bacteria.

CONCLUSION: The SEM technique is an excellent tool for differential recognition of the parasite surface morphology and molecular techniques could differentiate and identify Dirofilaria spp.}, } @article {pmid32488484, year = {2020}, author = {Semiatizki, A and Weiss, B and Bagim, S and Rohkin-Shalom, S and Kaltenpoth, M and Chiel, E}, title = {Effects, interactions, and localization of Rickettsia and Wolbachia in the house fly parasitoid, Spalangia endius.}, journal = {Microbial ecology}, volume = {80}, number = {3}, pages = {718-728}, doi = {10.1007/s00248-020-01520-x}, pmid = {32488484}, issn = {1432-184X}, support = {435/18//Planning and Budgeting Committee of the Council for Higher Education of Israel/ ; CoG 819585/ERC_/European Research Council/International ; }, mesh = {Animals ; Female ; *Host-Parasite Interactions ; Houseflies/parasitology ; Male ; Reproduction ; Rickettsia/*physiology ; *Symbiosis ; Wasps/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Many insect species harbor facultative microbial symbionts that affect their biology in diverse ways. Here, we studied the effects, interactions, and localization of two bacterial symbionts-Wolbachia and Rickettsia-in the parasitoid Spalangia endius. We crossed between four S. endius colonies-Wolbachia only (W), Rickettsia only (R), both (WR), and none (aposymbiotic, APS) (16 possible crosses) and found that Wolbachia induces incomplete cytoplasmic incompatibility (CI), both when the males are W or WR. Rickettsia did not cause reproductive manipulations and did not rescue the Wolbachia-induced CI. However, when R females were crossed with W or WR males, significantly less offspring were produced compared with that of control crosses. In non-CI crosses, the presence of Wolbachia in males caused a significant reduction in offspring numbers. Females' developmental time was significantly prolonged in the R colony, with adults starting to emerge one day later than the other colonies. Other fitness parameters did not differ significantly between the colonies. Using fluorescence in situ hybridization microscopy in females, we found that Wolbachia is localized alongside Rickettsia inside oocytes, follicle cells, and nurse cells in the ovaries. However, Rickettsia is distributed also in muscle cells all over the body, in ganglia, and even in the brain.}, } @article {pmid32485988, year = {2020}, author = {Papadopoulos, C and Karas, PA and Vasileiadis, S and Ligda, P and Saratsis, A and Sotiraki, S and Karpouzas, DG}, title = {Host Species Determines the Composition of the Prokaryotic Microbiota in Phlebotomus Sandflies.}, journal = {Pathogens (Basel, Switzerland)}, volume = {9}, number = {6}, pages = {}, pmid = {32485988}, issn = {2076-0817}, abstract = {Phlebotomine sandflies are vectors of the humans' and mammals' parasite Leishmania spp. Although the role of gut microbiome in the biological cycle of insects is acknowledged, we still know little about the factors modulating the composition of the gut microbiota of sandflies. We tested whether host species impose a strong structural effect on the gut microbiota of Phlebotomus spp. Sandflies were collected from the island of Leros, Greece, and classified to P. papatasi, P. neglectus, P. tobbi, and P. similis, all being negative to Leishmania spp. The prokaryotic gut microbiota was determined via 16S rRNA gene amplicon sequencing. Phlebotomus species supported distinct microbial communities (p < 0.001). P. papatasi microbiota was the most distinct over-dominated by three Spiroplasma, Wolbachia and Paenibacillus operational taxonomic units (OTUs), while another Wolbachia OTU prevailed in P. neglectus. Conversely, the microbiota of P. tobbi and P. similis was composed of several less dominant OTUs. Archaea showed low presence with the dominant OTUs belonging to methanogenic Euryarcheota, ammonia-oxidizing Thaumarcheota, and Nanoarchaeota. We provide first insights into the composition of the bacterial and archaeal community of Phlebotomus sandflies and showed that, in the absence of Leishmania, host genotype is the major modulator of Phlebotomus sandfly gut microbiota.}, } @article {pmid32477411, year = {2020}, author = {Mazzucco, R and Nolte, V and Vijayan, T and Schlötterer, C}, title = {Long-Term Dynamics Among Wolbachia Strains During Thermal Adaptation of Their Drosophila melanogaster Hosts.}, journal = {Frontiers in genetics}, volume = {11}, number = {}, pages = {482}, pmid = {32477411}, issn = {1664-8021}, support = {P 27630/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Climate change is a major evolutionary force triggering thermal adaptation in a broad range of species. While the consequences of global warming are being studied for an increasing number of species, limited attention has been given to the evolutionary dynamics of endosymbionts in response to climate change. Here, we address this question by studying the dynamics of Wolbachia, a well-studied endosymbiont of Drosophila melanogaster. D. melanogaster populations infected with 13 different Wolbachia strains were exposed to novel hot and cold laboratory environments for up to 180 generations. The short-term dynamics suggested a temperature-related fitness difference resulting in the increase of clade V strains in the cold environment only. Our long-term analysis now uncovers that clade V dominates in all replicates after generation 60 irrespective of temperature treatment. We propose that adaptation of the Drosophila host to either temperature or Drosophila C virus (DCV) infection are the cause of the replicated, temporally non-concordant Wolbachia dynamics. Our study provides an interesting case demonstrating that even simple, well-controlled experiments can result in complex, but repeatable evolutionary dynamics, thus providing a cautionary note on too simple interpretations on the impact of climate change.}, } @article {pmid32477320, year = {2020}, author = {Sharma, P and Rani, J and Chauhan, C and Kumari, S and Tevatiya, S and Das De, T and Savargaonkar, D and Pandey, KC and Dixit, R}, title = {Altered Gut Microbiota and Immunity Defines Plasmodium vivax Survival in Anopheles stephensi.}, journal = {Frontiers in immunology}, volume = {11}, number = {}, pages = {609}, pmid = {32477320}, issn = {1664-3224}, mesh = {Animals ; Anopheles/immunology/microbiology/*parasitology ; Gastrointestinal Microbiome/*physiology ; Plasmodium vivax/*growth & development ; RNA-Seq ; Symbiosis ; }, abstract = {Blood-feeding enriched gut-microbiota boosts mosquitoes' anti-Plasmodium immunity. Here, we ask how Plasmodium vivax alters gut-microbiota, anti-Plasmodial immunity, and impacts tripartite Plasmodium-mosquito-microbiota interactions in the gut lumen. We used a metagenomics and RNAseq strategy to address these questions. In naïve mosquitoes, Elizabethkingia meningitis and Pseudomonas spp. are the dominant bacteria and blood-feeding leads to a heightened detection of Elizabethkingia, Pseudomonas and Serratia 16S rRNA. A parallel RNAseq analysis of blood-fed midguts also shows the presence of Elizabethkingia-related transcripts. After, P. vivax infected blood-meal, however, we do not detect bacterial 16S rRNA until circa 36 h. Intriguingly, the transcriptional expression of a selected array of antimicrobial arsenal cecropins 1-2, defensin-1, and gambicin remained low during the first 36 h-a time frame when ookinetes/early oocysts invaded the gut. We conclude during the preinvasive phase, P. vivax outcompetes midgut-microbiota. This microbial suppression likely negates the impact of mosquito immunity which in turn may enhance the survival of P. vivax. Detection of sequences matching to mosquito-associated Wolbachia opens a new inquiry for its exploration as an agent for "paratransgenesis-based" mosquito control.}, } @article {pmid32473140, year = {2020}, author = {Burki, T}, title = {Wolbachia, a bacterium fighting on our side.}, journal = {The Lancet. Infectious diseases}, volume = {20}, number = {6}, pages = {662-663}, doi = {10.1016/S1473-3099(20)30384-4}, pmid = {32473140}, issn = {1474-4457}, mesh = {Aedes/*microbiology ; Animals ; Host-Pathogen Interactions ; Humans ; Malaria/*prevention & control/*transmission ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, } @article {pmid32471038, year = {2020}, author = {Tseng, SP and Hsu, PW and Lee, CC and Wetterer, JK and Hugel, S and Wu, LH and Lee, CY and Yoshimura, T and Yang, CS}, title = {Evidence for Common Horizontal Transmission of Wolbachia among Ants and Ant Crickets: Kleptoparasitism Added to the List.}, journal = {Microorganisms}, volume = {8}, number = {6}, pages = {}, pmid = {32471038}, issn = {2076-2607}, support = {N/A//Kyoto University/ ; }, abstract = {While Wolbachia, an intracellular bacterial symbiont, is primarily transmitted maternally in arthropods, horizontal transmission between species has been commonly documented. We examined kleptoparasitism as a potential mechanism for Wolbachia horizontal transmission, using ant crickets and their host ants as the model system. We compared prevalence and diversity of Wolbachia across multiple ant cricket species with different degrees of host specificity/integration level. Our analyses revealed at least three cases of inter-ordinal Wolbachia transfer among ant and ant crickets, and also showed that ant cricket species with high host-integration and host-specificity tend to harbor a higher Wolbachia prevalence and diversity than other types of ant crickets. This study provides empirical evidence that distribution of Wolbachia across ant crickets is largely attributable to horizontal transmission, but also elucidates the role of intimate ecological association in successful Wolbachia horizontal transmission.}, } @article {pmid32458175, year = {2020}, author = {Zhang, X and Liu, Q and Zhu, H}, title = {Modeling and dynamics of Wolbachia-infected male releases and mating competition on mosquito control.}, journal = {Journal of mathematical biology}, volume = {81}, number = {1}, pages = {243-276}, doi = {10.1007/s00285-020-01509-7}, pmid = {32458175}, issn = {1432-1416}, mesh = {*Aedes/microbiology ; Animals ; Humans ; Male ; *Models, Biological ; *Mosquito Control/methods/statistics & numerical data ; Mosquito Vectors/microbiology ; *Vector Borne Diseases/prevention & control ; *Wolbachia/physiology ; }, abstract = {Despite centuries of continuous efforts, mosquito-borne diseases (MBDs) remain enormous health threat of human life worldwide. Lately, the USA government has approved an innovative technology of releasing Wolbachia-infected male mosquitoes to suppress the wild mosquito population. In this paper we first introduce a stage-structured model for natural mosquitos, then we establish a new model considering the releasing of Wolbachia-infected male mosquitoes and the mating competition between the natural male mosquitoes and infected males on the suppression of natural mosquitoes. Dynamical analysis of the two models, including the existence and local stability of the equilibria and bifurcation analysis, reveals the existence of a forward bifurcation or a backward bifurcation with multiple attractors. Moreover, globally dynamical properties are further explored by using Lyapunov function and theory of monotone operators, respectively. Our findings suggest that infected male augmentation itself cannot always guarantee the success of population eradication, but leads to three possible levels of population suppression, so we define the corresponding suppression rate and estimate the minimum release ratio for population eradication. Furthermore, we study how the release ratio of infected males and natural ones, mating competition, the rate of cytoplasmic incompatibility and the basic offspring number affect the suppression rate of natural mosquitoes. Our results show that the successful eradication relies on assessing the reproductive capacity of natural mosquitoes, a selection of suitable Wolbachia strains and an appropriate release amount of infected males. This study will be helpful for public health authorities in designing proper strategies to control vector mosquitoes and prevent the epidemics of MBDs.}, } @article {pmid32456036, year = {2020}, author = {Yen, PS and Failloux, AB}, title = {A Review: Wolbachia-Based Population Replacement for Mosquito Control Shares Common Points with Genetically Modified Control Approaches.}, journal = {Pathogens (Basel, Switzerland)}, volume = {9}, number = {5}, pages = {}, pmid = {32456036}, issn = {2076-0817}, support = {n_ANR-10-LABX-62-IBEID//French Government's Investissement d'Avenir program, Laboratoire d'Excellence "Integrative Biology of Emerging Infectious Diseases, IBEID"/ ; NHRI-PP-108-0324-01-17-07//National Health Research Institutes/ ; }, abstract = {The growing expansion of mosquito vectors has made mosquito-borne arboviral diseases a global threat to public health, and the lack of licensed vaccines and treatments highlight the urgent need for efficient mosquito vector control. Compared to genetically modified control strategies, the intracellular bacterium Wolbachia, endowing a pathogen-blocking phenotype, is considered an environmentally friendly strategy to replace the target population for controlling arboviral diseases. However, the incomplete knowledge regarding the pathogen-blocking mechanism weakens the reliability of a Wolbachia-based population replacement strategy. Wolbachia infections are also vulnerable to environmental factors, temperature, and host diet, affecting their densities in mosquitoes and thus the virus-blocking phenotype. Here, we review the properties of the Wolbachia strategy as an approach to control mosquito populations in comparison with genetically modified control methods. Both strategies tend to limit arbovirus infections but increase the risk of selecting arbovirus escape mutants, rendering these strategies less reliable.}, } @article {pmid32455576, year = {2020}, author = {Laidoudi, Y and Marie, JL and Tahir, D and Watier-Grillot, S and Mediannikov, O and Davoust, B}, title = {Detection of Canine Vector-Borne Filariasis and Their Wolbachia Endosymbionts in French Guiana.}, journal = {Microorganisms}, volume = {8}, number = {5}, pages = {}, pmid = {32455576}, issn = {2076-2607}, abstract = {In French Guiana, canine heartworm disease is well known, but the diversity of filarial parasites of dogs remains largely unknown. A total of 98 canine blood samples from Cayenne and Kourou were assessed by a blood wet mount preparation, heartworm antigen test and molecular exploration of filarioid and Wolbachia DNAs, followed by a multiplex species-specific qPCR's identification and a subsequent sequencing analysis. Thereafter, a phylogeny based on maximum likelihood was carried out to facilitate specific identification. Five dogs were microfilaremic. Heartworm antigens were detected in 15 (15.3%) dogs. Of these, six (6.1%) were considered as occult infections as neither microfilariae nor Dirofilaria immitis DNA were detected. The 11 (11.2%) D. immitis isolates corresponded to a low virulent strain. Six of the D. immitis isolates were positive for Wolbachia endosymbionts of D. immitis belonging to the clade C DNA. Acanthocheilonema reconditum DNA was detected in 3 (3.1%) samples. Of these latter, one was found co-infected with the Brugia sp. genotype and the DNA of the clade D of the Wolbachia endosymbiont of Brugia species. This latter was also detected in two filarioid DNA-free samples. Finally, two samples were positive for Cercopithifilaria bainae genotype, which is distinct from those identified in Europe. The present study highlights the urgent need to implement chemoprophylaxis associated with anti-Wolbachia drugs to control these potential zoonoses.}, } @article {pmid32450914, year = {2020}, author = {Anders, KL and Indriani, C and Ahmad, RA and Tantowijoyo, W and Arguni, E and Andari, B and Jewell, NP and Dufault, SM and Ryan, PA and Tanamas, SK and Rancès, E and O'Neill, SL and Simmons, CP and Utarini, A}, title = {Update to the AWED (Applying Wolbachia to Eliminate Dengue) trial study protocol: a cluster randomised controlled trial in Yogyakarta, Indonesia.}, journal = {Trials}, volume = {21}, number = {1}, pages = {429}, pmid = {32450914}, issn = {1745-6215}, support = {R56 AI070043/AI/NIAID NIH HHS/United States ; R56 AI134724/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Dengue/epidemiology/*prevention & control/transmission/virology ; Dengue Virus/*pathogenicity ; Humans ; Incidence ; Indonesia/epidemiology ; Mosquito Vectors/*microbiology ; Pest Control, Biological/methods ; Randomized Controlled Trials as Topic ; Time Factors ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The AWED (Applying Wolbachia to Eliminate Dengue) trial is a parallel, two-arm, non-blinded cluster randomised controlled trial that is under way in Yogyakarta, Indonesia, with the aim of measuring the efficacy of Wolbachia-infected Aedes aegypti deployments in reducing dengue incidence in an endemic setting. Enrolment began in January 2018 and is ongoing. The original study protocol was published in April 2018. Here, we describe amendments that have been made to the study protocol since commencement of the trial.

METHODS: The key protocol amendments are (1) a revised study duration with planned end of participant enrolment in August 2020, (2) the addition of new secondary objectives (i) to estimate serotype-specific efficacy of the Wolbachia intervention and (ii) to compare Ae. aegypti abundance in intervention versus untreated clusters, (3) an additional exposure classification for the per-protocol analysis where the Wolbachia exposure index is calculated using only the cluster-level Wolbachia prevalence in the participant's cluster of residence, (4) power re-estimation using a multinomial sampling method that better accounts for randomness in sampling, and (5) the addition of two trial stopping rules to address the potential for persistently low rates of virologically confirmed dengue case enrolment and Wolbachia contamination into untreated clusters. Additional minor changes to the protocol are also described.

DISCUSSION: The findings from this study will provide the first experimental evidence for the efficacy of Wolbachia in reducing dengue incidence. Enrolment in the trial will conclude this year (2020) and results will be reported shortly thereafter.

TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT03055585. Registered on 14 February 2017. Last updated 22 March 2020.}, } @article {pmid32443976, year = {2020}, author = {Manzano-Marín, A}, title = {No evidence for Wolbachia as a nutritional co-obligate endosymbiont in the aphid Pentalonia nigronervosa.}, journal = {Microbiome}, volume = {8}, number = {1}, pages = {72}, pmid = {32443976}, issn = {2049-2618}, mesh = {Animals ; *Aphids ; *Buchnera/genetics ; Hemolymph ; Symbiosis ; *Wolbachia ; }, abstract = {Obligate symbiotic associations are present in a wide variety of animals with a nutrient-restricted diet. Aphids (hemiptera: Aphididae) almost-universally host Buchnera aphidicola bacteria in specialised organs (called bacteriomes). These bacteria supply the aphid with essential nutrients lacking from their diet (i.e. essential amino acids and some B vitamins). Some aphid lineages, such as species from the Lacninae subfamily, have evolved co-obligate associations with secondary endosymbionts, deriving from a loss of biotin- and riboflavin-biosynthetic genes. In this study, I re-analyse previously published sequencing data from the banana aphid Pentalonia nigronervosa. I show that the metabolic inference results from De Clerck et al. (Microbiome 3:63, 2015) are incorrect and possibly arise from the use of inadequate methods. Additionally, I discuss how the seemingly biased interpretation of their antibiotic treatment analyses together with an incorrect genome-based metabolic inference resulted in the erroneous suggestion "that a co-obligatory symbiosis between B. aphidicola and Wolbachia occurs in the banana aphid".}, } @article {pmid32442441, year = {2020}, author = {Górzyńska, K}, title = {Effects of the Clonostachys epichloë fungal hyperparasite on the symbiotic interaction between Botanophila flies and Epichloë fungus.}, journal = {Journal of invertebrate pathology}, volume = {174}, number = {}, pages = {107396}, doi = {10.1016/j.jip.2020.107396}, pmid = {32442441}, issn = {1096-0805}, mesh = {Animals ; Diptera/growth & development/*microbiology ; Epichloe/*physiology ; Holcus/growth & development ; *Host-Parasite Interactions ; Hypocreales/*physiology ; Larva/growth & development/microbiology ; Longevity ; Reproduction ; *Symbiosis ; }, abstract = {Botanophila flies are associated with Epichloë fungi in a symbiotic relationship in which the flies benefit from stromata as a food source for both adults and larvae, and the fungus benefits from the transfer of conidia by the flies, resulting in fertilization. Derogations from this pattern indicate that the Epichloë-Botanophila interaction cannot be clearly defined. The situation may be complicated by reports of new elements of the interaction, e.g., Wolbachia bacteria present in Botanophila larvae. The present study investigates the impact of Clonostachys epichloë (Speg.) Schroers, the fungal hyperparasite of Epichloë stromata, on the Botanophila-Epichloë interaction. The interaction between C. epichloë and Botanophila flies associated with Epichloë typhina subsp. clarkii (J.F. White) Leuchtm. & Schardl stromata was studied in the Holcus lanatus L. grass population. C. epichloë was present on 76.5% of stromata, covering on average 44.8 ± 32.1% of its surface and influencing the final perithecial coverage to the same extent as larval feeding. C. epichloë began to appear on stromata much later than the fly eggs and did not affect the preference for Botanophila egg laying. On the other hand, C. epichloë reduced larval hatching success and increased the mortality of the larvae. Clonostachys was responsible for 76.0% of all deaths, overgrowing brood chambers, and its mycelium was present both on and within larvae in all cases. Overall, as a result of the presence of C. epichloë, the number of Botanophila fly offspring decreased by 52.7%. Of the 26 surviving larvae, 10 (38.5%) were affected by C. epichloë, and their weight was significantly lower than that of unaffected larvae. Results show that C. epichloë, a new element of the interaction between E. typhina fungus and Botanophila flies, negatively affects both fungal reproduction and the offspring success of flies. This is the first report on the entomopathogenic activity of C. epichloë against Epichloë-associated Botanophila flies.}, } @article {pmid32413992, year = {2020}, author = {Ndii, MZ}, title = {Modelling the Use of Vaccine and Wolbachia on Dengue Transmission Dynamics.}, journal = {Tropical medicine and infectious disease}, volume = {5}, number = {2}, pages = {}, pmid = {32413992}, issn = {2414-6366}, support = {48/UN.17.19/PL/2019//Kementerian Pendidikan dan Kebudayaan/ ; }, abstract = {The use of vaccine and Wolbachia has been proposed as strategies against dengue. Research showed that the Wolbachia intervention is highly effective in areas with low to moderate transmission levels. On the other hand, the use of vaccine is strongly effective when it is implemented on seropositive individuals and areas with high transmission levels. The question that arises is could the combination of both strategies result in higher reduction in the number of dengue cases? This paper seeks to answer the aforementioned question by the use of a mathematical model. A deterministic model in the presence of vaccine and Wolbachia has been developed and analysed. Numerical simulations were presented and public health implications were discussed. The results showed that the performance of Wolbachia in reducing the number of dengue cases is better than that of vaccination if the vaccine efficacy is low, otherwise, the use of vaccine is sufficient to reduce dengue incidence and hence the combination of Wolbachia and vaccine is not necessary.}, } @article {pmid32406757, year = {2020}, author = {Nugent, CM and Elliott, TA and Ratnasingham, S and Adamowicz, SJ}, title = {coil: an R package for cytochrome c oxidase I (COI) DNA barcode data cleaning, translation, and error evaluation.}, journal = {Genome}, volume = {63}, number = {6}, pages = {291-305}, doi = {10.1139/gen-2019-0206}, pmid = {32406757}, issn = {1480-3321}, mesh = {Animals ; DNA Barcoding, Taxonomic/*methods ; DNA, Mitochondrial/genetics ; Electron Transport Complex IV/*genetics ; Frameshift Mutation/genetics ; Humans ; *Phylogeny ; Pseudogenes/*genetics ; }, abstract = {Biological conclusions based on DNA barcoding and metabarcoding analyses can be strongly influenced by the methods utilized for data generation and curation, leading to varying levels of success in the separation of biological variation from experimental error. The 5' region of cytochrome c oxidase subunit I (COI-5P) is the most common barcode gene for animals, with conserved structure and function that allows for biologically informed error identification. Here, we present coil (https://CRAN.R-project.org/package=coil), an R package for the pre-processing and frameshift error assessment of COI-5P animal barcode and metabarcode sequence data. The package contains functions for placement of barcodes into a common reading frame, accurate translation of sequences to amino acids, and highlighting insertion and deletion errors. The analysis of 10 000 barcode sequences of varying quality demonstrated how coil can place barcode sequences in reading frame and distinguish sequences containing indel errors from error-free sequences with greater than 97.5% accuracy. Package limitations were tested through the analysis of COI-5P sequences from the plant and fungal kingdoms as well as the analysis of potential contaminants: nuclear mitochondrial pseudogenes and Wolbachia COI-5P sequences. Results demonstrated that coil is a strong technical error identification method but is not reliable for detecting all biological contaminants.}, } @article {pmid32402821, year = {2020}, author = {Zumaquero, L and Simón, F and Carretón, E and Hernández, I and Sandoval, C and Morchón, R}, title = {Prevalence of canine and human dirofilariosis in Puebla, Mexico.}, journal = {Veterinary parasitology}, volume = {282}, number = {}, pages = {109098}, doi = {10.1016/j.vetpar.2020.109098}, pmid = {32402821}, issn = {1873-2550}, mesh = {Adolescent ; Adult ; Aged ; Animals ; Child ; Child, Preschool ; Dirofilaria immitis/*isolation & purification ; Dirofilariasis/*epidemiology/parasitology ; Dog Diseases/*epidemiology/parasitology ; Dogs ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mexico/epidemiology ; Microfilariae/isolation & purification ; Middle Aged ; Prevalence ; Risk Factors ; Seroepidemiologic Studies ; Young Adult ; }, abstract = {The objective of this work was to determine the risk of exposure to Dirofilaria immitis in the stray dog population and people living in the metropolitan area of Puebla City, Mexico, using serologic methods. A total of 283 blood salmples were collected from dogs and 254 salmples from people. The canine samples were analyzed using a commercial kit to detect D. immitis antigens and the Knott test to detect microfilariae. Human samples were analyzed by ELISAs to detect antibodies against D. immitis and Wolbachia anti-WSP, as well as Western blot to identify specific bands of the parasite in the adult antigenic extract of D. immitis. Positive results were represented on a GIS map that included layers with geoenvironmental information relevant to parasite transmission in the study area. The prevalence of D. immitis in the canine stray population was 2.12 %, with positive cases found in individuals aged 1-10 years. Human seroprevalence was established at 5.11 %, with women and men similarly affected. By age, a significantly higher seroprevalence was observed in younger individuals (<18 years) than in the rest of the population. The geolocation of the results showed that positive cases were found in the environment (within 2 km) of potentially favorable areas for mosquito breeding. The importance of human influence in the creation and maintenance of favorable conditions for the transmission of canine and human dirofilariosis within an urban area is highlighted, both by the lack of environmental sanitation and by the absence of measures to limit or eliminate the canine reservoir population.}, } @article {pmid32391935, year = {2020}, author = {Kaur, R and Martinez, J and Rota-Stabelli, O and Jiggins, FM and Miller, WJ}, title = {Age, tissue, genotype and virus infection regulate Wolbachia levels in Drosophila.}, journal = {Molecular ecology}, volume = {29}, number = {11}, pages = {2063-2079}, doi = {10.1111/mec.15462}, pmid = {32391935}, issn = {1365-294X}, support = {P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {*Age Factors ; Animals ; *Drosophila/genetics/microbiology/virology ; Genotype ; *Symbiosis/genetics ; *Virus Diseases ; *Wolbachia ; }, abstract = {The bacterial symbiont Wolbachia can protect insects against viral pathogens, and the varying levels of antiviral protection are correlated with the endosymbiont load within the insects. To understand why Wolbachia strains differ in their antiviral effects, we investigated the factors controlling Wolbachia density in five closely related strains in their natural Drosophila hosts. We found that Wolbachia density varied greatly across different tissues and between flies of different ages, and these effects depended on the host-symbiont association. Some endosymbionts maintained largely stable densities as flies aged while others increased, and these effects in turn depended on the tissue being examined. Measuring Wolbachia rRNA levels in response to viral infection, we found that viral infection itself also altered Wolbachia levels, with Flock House virus causing substantial reductions in symbiont loads late in the infection. This effect, however, was virus-specific as Drosophila C virus had little impact on Wolbachia in all of the five host systems. Because viruses have strong tissue tropisms and antiviral protection is thought to be cell-autonomous, these effects are likely to affect the virus-blocking phenomenon. However, we were unable to find any evidence of a correlation between Wolbachia and viral titres within the same tissues. We conclude that Wolbachia levels within flies are regulated in a complex host-symbiont-virus-dependent manner and this trinity is likely to influence the antiviral effects of Wolbachia.}, } @article {pmid32388920, year = {2020}, author = {Zhu, YX and Song, ZR and Song, YL and Hong, XY}, title = {Double infection of Wolbachia and Spiroplasma alters induced plant defense and spider mite fecundity.}, journal = {Pest management science}, volume = {76}, number = {9}, pages = {3273-3281}, doi = {10.1002/ps.5886}, pmid = {32388920}, issn = {1526-4998}, support = {2019M651864//China Postdoctoral Science Foundation/ ; 31672035//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; 1901888//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Fertility ; *Mites ; *Spiroplasma ; *Tetranychidae ; *Wolbachia ; }, abstract = {BACKGROUND: Herbivore-associated bacterial symbionts can change plant physiology and influence herbivore fitness. The spider mite Tetranychus truncatus is a notorious pest harboring various bacterial symbionts; however, the effect of bacterial symbionts on host plant physiology remains unclear. Here, we investigated whether infection with the endosymbionts Wolbachia and Spiroplasma altered spider mite performance on tomato plants and affected plant-induced defenses.

RESULTS: Wolbachia and Spiroplasma were mainly located in the gnathosoma and ovaries of their spider mite hosts. Wolbachia and Spiroplasma significantly improved spider mite reproductive performance in cultivated and wild-type tomato. However, in plants deficient in jasmonic acid (JA) and salicylic acid (SA), there were no significant differences in reproduction between spider mites infected with Wolbachia and Spiroplasma and uninfected mites. The results indicated that the reproduction benefits conferred by endosymbionts may relate to plant defenses. Both spider mites infected with Wolbachia and Spiroplasma and uninfected mites induced similar levels of JA and SA accumulation in tomato, whereas tomato plants damaged by spider mites infected with both Wolbachia and Spiroplasma showed lower expression levels of JA- and SA-responsive genes than those damaged by uninfected spider mites. In addition, mites infected with Wolbachia and Spiroplasma mites consumed more tomato amino acids compared to uninfected spider mites, which may have contributed to host fecundity.

CONCLUSIONS: Our results suggest that the reproduction benefits conferred by endosymbionts may be associated with changes in plant defense parameters and the concentrations of plant amino acids. The results highlight the importance of endosymbionts in interactions between spider mites and their host plants. © 2020 Society of Chemical Industry.}, } @article {pmid32384776, year = {2020}, author = {Bryant, KN and Newton, ILG}, title = {The Intracellular Symbiont Wolbachia pipientis Enhances Recombination in a Dose-Dependent Manner.}, journal = {Insects}, volume = {11}, number = {5}, pages = {}, pmid = {32384776}, issn = {2075-4450}, support = {1456545//National Science Foundation/ ; }, abstract = {Wolbachia pipientis is an intracellular alphaproteobacterium that infects 40%-60% of insect species and is well known for host reproductive manipulations. Although Wolbachia are primarily maternally transmitted, evidence of horizontal transmission can be found in incongruent host-symbiont phylogenies and recent acquisitions of the same Wolbachia strain by distantly related species. Parasitoids and predator-prey interactions may indeed facilitate the transfer of Wolbachia between insect lineages, but it is likely that Wolbachia are acquired via introgression in many cases. Many hypotheses exist to explain Wolbachia prevalence and penetrance, such as nutritional supplementation, protection from parasites, protection from viruses, or direct reproductive parasitism. Using classical genetics, we show that Wolbachia increase recombination in infected lineages across two genomic intervals. This increase in recombination is titer-dependent as the wMelPop variant, which infects at higher load in Drosophila melanogaster, increases recombination 5% more than the wMel variant. In addition, we also show that Spiroplasma poulsonii, another bacterial intracellular symbiont of D. melanogaster, does not induce an increase in recombination. Our results suggest that Wolbachia infection specifically alters its host's recombination landscape in a dose-dependent manner.}, } @article {pmid32358830, year = {2020}, author = {Sandoval-Mojica, AF and Altman, S and Hunter, WB and Pelz-Stelinski, KS}, title = {Peptide conjugated morpholinos for management of the huanglongbing pathosystem.}, journal = {Pest management science}, volume = {76}, number = {9}, pages = {3217-3224}, doi = {10.1002/ps.5877}, pmid = {32358830}, issn = {1526-4998}, support = {2016-70016-24782//National Institute of Food and Agriculture/ ; }, mesh = {Animals ; *Citrus ; *Hemiptera ; Morpholinos ; Peptides ; Plant Diseases ; *Rhizobiaceae/genetics ; }, abstract = {BACKGROUND: 'Candidatus Liberibacter asiaticus' (CLas) is the causal agent of the devastating citrus disease Huanglongbing (HLB) and is transmitted by the insect vector Diaphorina citri (Hemiptera: Liviidae). A potential approach for treating CLas infection is the use of synthetic nucleic acid-like oligomers to silence bacterial gene expression. Peptide conjugated morpholinos (PPMOs) targeting essential genes in CLas and the psyllid vector's endosymbiotic bacteria, Wolbachia (-Diaphorina, wDi), were evaluated using in vitro and in vivo assays.

RESULTS: Expression of the wDi gyrA gene was significantly reduced following incubation of wDi cells with PPMOs. In addition, the viability of isolated wDi cells was greatly reduced when treated with PPMOs as compared to untreated cells. Feeding D. citri adults with a complementary PPMO (CLgyrA-14) showed significantly reduced (70% lower) expression of the CLas gyrA gene. CLas relative density was significantly lower in the psyllids fed with CLgyrA-14, when compared to untreated insects. Psyllids that were treated with CLgyrA-14 were less successful in transmitting the pathogen into uninfected plants, compared to untreated insects.

CONCLUSION: The expression of essential genes in the D. citri symbiont, wDi and the HLB pathogen were suppressed in response to PPMO treatments. This study demonstrates the potential of PPMOs as a novel strategy for management of bacterial pathogens of fruit trees, such as HLB. © 2020 Society of Chemical Industry.}, } @article {pmid32340230, year = {2020}, author = {Dahmana, H and Mediannikov, O}, title = {Mosquito-Borne Diseases Emergence/Resurgence and How to Effectively Control It Biologically.}, journal = {Pathogens (Basel, Switzerland)}, volume = {9}, number = {4}, pages = {}, pmid = {32340230}, issn = {2076-0817}, abstract = {Deadly pathogens and parasites are transmitted by vectors and the mosquito is considered the most threatening vector in public health, transmitting these pathogens to humans and animals. We are currently witnessing the emergence/resurgence in new regions/populations of the most important mosquito-borne diseases, such as arboviruses and malaria. This resurgence may be the consequence of numerous complex parameters, but the major cause remains the mismanagement of insecticide use and the emergence of resistance. Biological control programmes have rendered promising results but several highly effective techniques, such as genetic manipulation, remain insufficiently considered as a control mechanism. Currently, new strategies based on attractive toxic sugar baits and new agents, such as Wolbachia and Asaia, are being intensively studied for potential use as alternatives to chemicals. Research into new insecticides, Insect Growth Regulators, and repellent compounds is pressing, and the improvement of biological strategies may provide key solutions to prevent outbreaks, decrease the danger to at-risk populations, and mitigate resistance.}, } @article {pmid32339445, year = {2021}, author = {Lv, N and Peng, J and Chen, XY and Guo, CF and Sang, W and Wang, XM and Ahmed, MZ and Xu, YY and Qiu, BL}, title = {Antagonistic interaction between male-killing and cytoplasmic incompatibility induced by Cardinium and Wolbachia in the whitefly, Bemisia tabaci.}, journal = {Insect science}, volume = {28}, number = {2}, pages = {330-346}, doi = {10.1111/1744-7917.12793}, pmid = {32339445}, issn = {1744-7917}, support = {2018B020205003//Key-Area Research and Development Program of Guangdong Province/ ; 31672028//National Natural Science Foundation of China/ ; U1701231//NSFC-Guangdong Joint Research Fund/ ; }, mesh = {Animals ; Bacteroidetes/*physiology ; Female ; Fertility ; Hemiptera/*microbiology/physiology ; Longevity ; Male ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Cardinium and Wolbachia are maternally inherited bacterial symbionts of arthropods that can manipulate host reproduction by increasing the fitness of infected females. Here, we report that Cardinium and Wolbachia coinfection induced male-killing and cytoplasmic incompatibility (CI) when they coexisted in a cryptic species of whitefly, Bemisia tabaci Asia II7. Cardinium and Wolbachia symbionts were either singly or simultaneously localized in the bacteriocytes placed in the abdomen of B. tabaci nymphs and adults. Cardinium-Wolbachia coinfection induced male-killing and resulted in a higher female sex ratio in the intraspecific amphigenetic progeny of Asia II7 ICWH and ICWL lines; interestingly, male-killing induction was enhanced with increased Cardinium titer. Moreover, single infection of Wolbachia induced partial CI in the Asia II7 IW line and resulted in reduced fecundity, higher embryonic mortality, and lower female sex ratio. The uninfected Asia II7 IU line had significantly higher fecundity, lower embryonic and nymphal mortalities, and a lower level of CI than both the Wolbachia-infected Asia II7 IW line and the Cardinium-Wolbachia-coinfected Asia II7 ICWH line. Our findings indicate that Cardinium-Wolbachia coinfection induced male-killing, which may have had antagonistic effects on Wolbachia-induced CI in the Asia II7 whiteflies. For the first time, our study revealed that B. tabaci Asia II7 reproduction is co-manipulated by Cardinium and Wolbachia endosymbionts.}, } @article {pmid32314037, year = {2020}, author = {Yang, J and Wei, S and Su, D and Zhang, Z and Chen, S and Luo, Z and Shen, X and Lai, Y and Jamil, A and Tong, J and Cui, X}, title = {Comparison of the Rhizosphere Soil Microbial Community Structure and Diversity Between Powdery Mildew-Infected and Noninfected Strawberry Plants in a Greenhouse by High-Throughput Sequencing Technology.}, journal = {Current microbiology}, volume = {77}, number = {8}, pages = {1724-1736}, pmid = {32314037}, issn = {1432-0991}, support = {31660089//The National Natural Science Foundation of China/ ; 31960220//The National Natural Science Foundation of China/ ; }, mesh = {Archaea/classification ; Bacteria/classification ; Biodiversity ; Fragaria/*microbiology ; High-Throughput Nucleotide Sequencing ; *Microbiota ; Plant Diseases/*microbiology ; Plant Roots/microbiology ; RNA, Ribosomal, 16S/genetics ; *Rhizosphere ; *Soil Microbiology ; }, abstract = {The aim of this study was to compare the microbial community structure and diversity in powdery mildew-infected and noninfected strawberry plant rhizosphere soils in the greenhouse based on variations in the 16S rRNA gene V3-V4 and fungal ITS2 regions by Illumina amplicon sequencing. Powdery mildew infection reduced the number of operational taxonomic units (OTUs) and prokaryotic and fungal community richness/diversity indexes in the rhizosphere soils compared with those in healthy plant soils. Furthermore, 3543 prokaryotic and 581 fungal OTUs were obtained at the 97% similarity level. Proteobacteria, Actinobacteria, Bacteroidetes, Acidobacteria, and Chloroflexi were the dominant bacterial phyla; Woesearchaeota_DHVEG-6, Bathyarchaeota, and Thaumarchaeota were the dominant archaea; and Ascomycota, Basidiomycota, unclassified_fungi, and Zygomycota were the dominant fungal phyla. Their proportions differed significantly among samples. Wolbachia, Devosia, Pseudolabrys, Streptomyces, and Rhizomicrobium were the most abundant bacterial genera; their proportions differed significantly among samples. Most Pseudomonas, Streptomyces, and 'norank' group members might be potential antagonistic microorganisms of powdery mildew pathogens, and Wolbachia and Rickettsia might be pathogen-transmitting vectors. Microascus, Clitopilus, and Ciliophora were the dominant fungi, and their community structures and abundances significantly differed among samples. Microascus, Talaromyces, Zopfiella, and Cryptococcus were relatively more abundant in the powdery mildew-infected strawberry plant rhizosphere soils. Fusarium, Trichoderma, Clitopilus, and 'unclassified' group members may be potential antagonistic populations. The results suggested that powdery mildew-infected strawberry fruits and plants cannot be consumed. This report is the first study to illustrate differences in the rhizosphere soil prokaryotic and fungal communities between powdery mildew-infected and noninfected strawberry plants in a greenhouse.}, } @article {pmid32314003, year = {2020}, author = {Parker, ES and Newton, ILG and Moczek, AP}, title = {(My Microbiome) Would Walk 10,000 miles: Maintenance and Turnover of Microbial Communities in Introduced Dung Beetles.}, journal = {Microbial ecology}, volume = {80}, number = {2}, pages = {435-446}, doi = {10.1007/s00248-020-01514-9}, pmid = {32314003}, issn = {1432-184X}, support = {1901680//Division of Integrative Organismal Systems/ ; 1256689//Division of Integrative Organismal Systems/ ; 61369//John Templeton Foundation/ ; }, mesh = {Animals ; Australian Capital Territory ; Bacteria/*isolation & purification ; Coleoptera/*microbiology ; Introduced Species ; Italy ; *Microbiota ; *Symbiosis ; West Virginia ; }, abstract = {Host-associated microbes facilitate diverse biotic and abiotic interactions between hosts and their environments. Experimental alterations of host-associated microbial communities frequently decrease host fitness, yet much less is known about if and how host-microbiome interactions are altered by natural perturbations, such as introduction events. Here, we begin to assess this question in Onthophagus dung beetles, a species-rich and geographically widely distributed genus whose members rely on vertically transmitted microbiota to support normal development. Specifically, we investigated to what extent microbiome community membership shifts during host introduction events and the relative significance of ancestral associations and novel environmental conditions in the structuring of microbial communities of introduced host species. Our results demonstrate that both evolutionary history and local environmental forces structure the microbial communities of these animals, but that their relative importance is shaped by the specific circumstances that characterize individual introduction events. Furthermore, we identify microbial taxa such as Dysgonomonas that may constitute members of the core Onthophagus microbiome regardless of host population or species, but also Wolbachia which associates with Onthophagus beetles in a species or even population-specific manner. We discuss the implications of our results for our understanding of the evolutionary ecology of symbiosis in dung beetles and beyond.}, } @article {pmid31667465, year = {2019}, author = {Ryan, PA and Turley, AP and Wilson, G and Hurst, TP and Retzki, K and Brown-Kenyon, J and Hodgson, L and Kenny, N and Cook, H and Montgomery, BL and Paton, CJ and Ritchie, SA and Hoffmann, AA and Jewell, NP and Tanamas, SK and Anders, KL and Simmons, CP and O'Neill, SL}, title = {Establishment of wMel Wolbachia in Aedes aegypti mosquitoes and reduction of local dengue transmission in Cairns and surrounding locations in northern Queensland, Australia.}, journal = {Gates open research}, volume = {3}, number = {}, pages = {1547}, pmid = {31667465}, issn = {2572-4754}, support = {/WT_/Wellcome Trust/United Kingdom ; R56 AI134724/AI/NIAID NIH HHS/United States ; }, abstract = {Background: The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and subsequently shown in laboratory studies to reduce transmission of a range of viruses including dengue, Zika, chikungunya, yellow fever, and Mayaro viruses that cause human disease. Here we report the entomological and epidemiological outcomes of staged deployment of Wolbachia across nearly all significant dengue transmission risk areas in Australia. Methods: The wMel strain of Wolbachia was backcrossed into the local Aedes aegypti genotype (Cairns and Townsville backgrounds) and mosquitoes were released in the field by staff or via community assisted methods. Mosquito monitoring was undertaken and mosquitoes were screened for the presence of Wolbachia. Dengue case notifications were used to track dengue incidence in each location before and after releases. Results: Empirical analyses of the Wolbachia mosquito releases, including data on the density, frequency and duration of Wolbachia mosquito releases, indicate that Wolbachia can be readily established in local mosquito populations, using a variety of deployment options and over short release durations (mean release period 11 weeks, range 2-22 weeks). Importantly, Wolbachia frequencies have remained stable in mosquito populations since releases for up to 8 years. Analysis of dengue case notifications data demonstrates near-elimination of local dengue transmission for the past five years in locations where Wolbachia has been established. The regression model estimate of Wolbachia intervention effect from interrupted time series analyses of case notifications data prior to and after releases, indicated a 96% reduction in dengue incidence in Wolbachia treated populations (95% confidence interval: 84 - 99%). Conclusion: Deployment of the wMel strain of Wolbachia into local Ae. aegypti populations across the Australian regional cities of Cairns and most smaller regional communities with a past history of dengue has resulted in the reduction of local dengue transmission across all deployment areas.}, } @article {pmid32307514, year = {2020}, author = {Lau, MJ and Ross, PA and Endersby-Harshman, NM and Hoffmann, AA}, title = {Impacts of Low Temperatures on Wolbachia (Rickettsiales: Rickettsiaceae)-Infected Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {57}, number = {5}, pages = {1567-1574}, pmid = {32307514}, issn = {1938-2928}, support = {108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/physiology ; Animals ; *Cold Temperature ; Female ; Fertility ; Wolbachia/*virology ; }, abstract = {In recent decades, the occurrence and distribution of arboviral diseases transmitted by Aedes aegypti mosquitoes has increased. In a new control strategy, populations of mosquitoes infected with Wolbachia are being released to replace existing populations and suppress arboviral disease transmission. The success of this strategy can be affected by high temperature exposure, but the impact of low temperatures on Wolbachia-infected Ae. aegypti is unclear, even though low temperatures restrict the abundance and distribution of this species. In this study, we considered low temperature cycles relevant to the spring season that are close to the distribution limits of Ae. aegypti, and tested the effects of these temperature cycles on Ae. aegypti, Wolbachia strains wMel and wAlbB, and Wolbachia phage WO. Low temperatures influenced Ae. aegypti life-history traits, including pupation, adult eclosion, and fertility. The Wolbachia-infected mosquitoes, especially wAlbB, performed better than uninfected mosquitoes. Temperature shift experiments revealed that low temperature effects on life history and Wolbachia density depended on the life stage of exposure. Wolbachia density was suppressed at low temperatures but densities recovered with adult age. In wMel Wolbachia there were no low temperature effects specific to Wolbachia phage WO. The findings suggest that Wolbachia-infected Ae. aegypti are not adversely affected by low temperatures, indicating that the Wolbachia replacement strategy is suitable for areas experiencing cool temperatures seasonally.}, } @article {pmid32302295, year = {2020}, author = {Tantowijoyo, W and Andari, B and Arguni, E and Budiwati, N and Nurhayati, I and Fitriana, I and Ernesia, I and Daniwijaya, EW and Supriyati, E and Yusdiana, DH and Victorius, M and Wardana, DS and Ardiansyah, H and Ahmad, RA and Ryan, PA and Simmons, CP and Hoffmann, AA and Rancès, E and Turley, AP and Johnson, P and Utarini, A and O'Neill, SL}, title = {Stable establishment of wMel Wolbachia in Aedes aegypti populations in Yogyakarta, Indonesia.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {4}, pages = {e0008157}, pmid = {32302295}, issn = {1935-2735}, mesh = {Aedes/*microbiology/virology ; Animals ; Arboviruses ; Australia ; Biological Control Agents ; Disease Transmission, Infectious/prevention & control ; Female ; Indonesia ; Male ; Mosquito Vectors/*microbiology ; Wolbachia/*physiology ; }, abstract = {The successful establishment of the wMel strain of Wolbachia for the control of arbovirus transmission by Aedes aegypti has been proposed and is being implemented in a number of countries. Here we describe the successful establishment of the wMel strain of Wolbachia in four sites in Yogyakarta, Indonesia. We demonstrate that Wolbachia can be successfully introgressed after transient releases of wMel-infected eggs or adult mosquitoes. We demonstrate that the approach is acceptable to communities and that Wolbachia maintains itself in the mosquito population once deployed. Finally, our data show that spreading rates of Wolbachia in the Indonesian setting are slow which may reflect more limited dispersal of Aedes aegypti than seen in other sites such as Cairns, Australia.}, } @article {pmid32299043, year = {2020}, author = {Mendiola, SY and Civitello, DJ and Gerardo, NM}, title = {An integrative approach to symbiont-mediated vector control for agricultural pathogens.}, journal = {Current opinion in insect science}, volume = {39}, number = {}, pages = {57-62}, doi = {10.1016/j.cois.2020.02.007}, pmid = {32299043}, issn = {2214-5753}, mesh = {Animals ; Crops, Agricultural/*microbiology ; Gene Silencing ; Insect Control/*methods ; Insect Vectors/*microbiology ; Plant Diseases/microbiology/prevention & control ; RNA Interference ; Vector Borne Diseases/*microbiology/prevention & control ; Wolbachia ; }, abstract = {Vector-borne pathogens pose significant threats to agricultural productivity. Methods that exploit associations between insects and their symbiotic microbes, dubbed symbiont-mediated vector control, are emerging as viable alternatives to insecticides for the control of vector-borne agricultural plant pathogens. The development of methods for effective microbial manipulation, such as RNA interference and paratransgenesis, may facilitate symbiont-mediated vector control tactics aimed at either suppressing insect populations or at manipulating vector competence, an insect vector's ability to acquire, harbor, and transmit pathogens. As suppression strategies transition from the laboratory to the field, the need for methods to evaluate their viability and predict their outcomes is apparent. Mathematical models of symbiont impact on agricultural disease can inform the development of symbiont-mediated vector control. We propose an integrative approach, combining theoretical and empirical experiments to identify the best practices for achieving meaningful improvements to crop health and productivity.}, } @article {pmid32298447, year = {2020}, author = {Seifert, B}, title = {The Gene and Gene Expression (GAGE) Species Concept: An Universal Approach for All Eukaryotic Organisms.}, journal = {Systematic biology}, volume = {69}, number = {5}, pages = {1033-1038}, doi = {10.1093/sysbio/syaa032}, pmid = {32298447}, issn = {1076-836X}, mesh = {Classification/*methods ; Eukaryota/*classification/*genetics ; *Gene Expression ; Genes/*genetics ; *Genetic Speciation ; }, abstract = {The Gene and Gene Expression (GAGE) species concept, a new version of the Pragmatic Species Concept of Seifert (2014), is proposed as a concept applicable to any described recent or fossil eukaryotic organism independent from its mode of reproduction or evolutionary history. In addition to presenting the concept as such, the article also provides practical recommendations for taxonomists when delimiting species and describing taxa. The wording of the new concept contains a heading core sentence plus five attached sentences addressing essential conditions for its translation into a sound taxonomic practice: "Species are separable clusters that have passed a threshold of evolutionary divergence and are exclusively defined by nuclear DNA sequences and/or their expression products. Nuclear DNA sequences and their expression products are different character systems but have a highly correlated indicative function. Character systems with the least risk of epigenetic or ontogenetic modification have superior indicative value when conflicts between character systems of integrative studies arise. All character systems have to be described by an adequate numerics allowing cluster formation and determination of thresholds. Thresholds for each character system should be fixed by consensus among the experts under the principle of avoiding oversplitting or lumping. Clusters must not be the expression of intraspecific polymorphism." Recognizing the distortions and conflicts caused to taxonomy through barcoding or through assessment on the basis of association with other organisms, the GAGE species concept strongly downgrades the use of cytoplasmic DNA of endosymbiotic origin (mtDNA, cpDNA) or DNA of closely associated microbes (e.g., Wolbachia bacteria) for final taxonomic decision-making. Recognizing the distortion of phylogenies by the high frequency of reticulate evolution, it is argued that delimiting and naming species has to be separated from constructing bifurcating phylogenetic trees. [Cytoplasmic DNA; lumping; nuclear DNA; numeric taxonomy; oversplitting; reticulate evolution.].}, } @article {pmid32282862, year = {2020}, author = {Flores, HA and Taneja de Bruyne, J and O'Donnell, TB and Tuyet Nhu, V and Thi Giang, N and Thi Xuan Trang, H and Thi Thuy Van, H and Thi Long, V and Thi Dui, L and Le Anh Huy, H and Thi Le Duyen, H and Thi Van Thuy, N and Thanh Phong, N and Van Vinh Chau, N and Thi Hue Kien, D and Thuy Vi, T and Wills, B and O'Neill, SL and Simmons, CP and Carrington, LB}, title = {Multiple Wolbachia strains provide comparative levels of protection against dengue virus infection in Aedes aegypti.}, journal = {PLoS pathogens}, volume = {16}, number = {4}, pages = {e1008433}, pmid = {32282862}, issn = {1553-7374}, support = {102591/Z/13/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/genetics/metabolism/*microbiology/*virology ; Animals ; Dengue Virus/*physiology ; Female ; Male ; Mosquito Vectors/genetics/metabolism/*microbiology/*virology ; Pest Control, Biological ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {The insect bacterium Wolbachia pipientis is being introgressed into Aedes aegypti populations as an intervention against the transmission of medically important arboviruses. Here we compare Ae. aegypti mosquitoes infected with wMelCS or wAlbB to the widely used wMel Wolbachia strain on an Australian nuclear genetic background for their susceptibility to infection by dengue virus (DENV) genotypes spanning all four serotypes. All Wolbachia-infected mosquitoes were more resistant to intrathoracic DENV challenge than their wildtype counterparts. Blocking of DENV replication was greatest by wMelCS. Conversely, wAlbB-infected mosquitoes were more susceptible to whole body infection than wMel and wMelCS. We extended these findings via mosquito oral feeding experiments, using viremic blood from 36 acute, hospitalised dengue cases in Vietnam, additionally including wMel and wildtype mosquitoes on a Vietnamese nuclear genetic background. As above, wAlbB was less effective at blocking DENV replication in the abdomen compared to wMel and wMelCS. The transmission potential of all Wolbachia-infected mosquito lines (measured by the presence/absence of infectious DENV in mosquito saliva) after 14 days, was significantly reduced compared to their wildtype counterparts, and lowest for wMelCS and wAlbB. These data support the use of wAlbB and wMelCS strains for introgression field trials and the biocontrol of DENV transmission. Furthermore, despite observing significant differences in transmission potential between wildtype mosquitoes from Australia and Vietnam, no difference was observed between wMel-infected mosquitoes from each background suggesting that Wolbachia may override any underlying variation in DENV transmission potential.}, } @article {pmid32278312, year = {2020}, author = {Patterson, EI and Villinger, J and Muthoni, JN and Dobel-Ober, L and Hughes, GL}, title = {Exploiting insect-specific viruses as a novel strategy to control vector-borne disease.}, journal = {Current opinion in insect science}, volume = {39}, number = {}, pages = {50-56}, pmid = {32278312}, issn = {2214-5753}, support = {R21 AI129507/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; BB/T001240/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Aedes/virology ; Animals ; Arboviruses/growth & development ; Biological Control Agents/isolation & purification ; Flavivirus/genetics/isolation & purification/physiology ; Flavivirus Infections/prevention & control/transmission ; Insect Control/*methods ; *Insect Viruses/genetics/isolation & purification/physiology ; Insecta/virology ; Mosquito Vectors/*virology ; Phylogeny ; Vector Borne Diseases/*prevention & control/transmission ; }, abstract = {Novel insect-specific viruses (ISVs) are being discovered in many important vectors due to advances in sequencing technology and a growing awareness of the virome. Several in vitro and in vivo studies indicate that ISVs are capable of modulating pathogenic arboviruses. In addition, there is growing evidence that both vertical and horizonal transmission strategies maintain ISVs in vector populations. As such there is potential to exploit ISVs for stand-alone vector control strategies and deploying them in synergy with other symbiont control approaches such as Wolbachia-mediated control. However, before the applied potential can be realized, a greater understanding of their basic biology is required, including their species range, ability to be maintained and transmitted in native and non-native vector hosts, and the effect of infection on a range of pathogens.}, } @article {pmid32265562, year = {2020}, author = {Crawford, JE and Clarke, DW and Criswell, V and Desnoyer, M and Cornel, D and Deegan, B and Gong, K and Hopkins, KC and Howell, P and Hyde, JS and Livni, J and Behling, C and Benza, R and Chen, W and Dobson, KL and Eldershaw, C and Greeley, D and Han, Y and Hughes, B and Kakani, E and Karbowski, J and Kitchell, A and Lee, E and Lin, T and Liu, J and Lozano, M and MacDonald, W and Mains, JW and Metlitz, M and Mitchell, SN and Moore, D and Ohm, JR and Parkes, K and Porshnikoff, A and Robuck, C and Sheridan, M and Sobecki, R and Smith, P and Stevenson, J and Sullivan, J and Wasson, B and Weakley, AM and Wilhelm, M and Won, J and Yasunaga, A and Chan, WC and Holeman, J and Snoad, N and Upson, L and Zha, T and Dobson, SL and Mulligan, FS and Massaro, P and White, BJ}, title = {Efficient production of male Wolbachia-infected Aedes aegypti mosquitoes enables large-scale suppression of wild populations.}, journal = {Nature biotechnology}, volume = {38}, number = {4}, pages = {482-492}, pmid = {32265562}, issn = {1546-1696}, mesh = {Aedes/growth & development/*microbiology/*physiology ; Animal Migration ; Animals ; California ; Female ; Larva/growth & development/microbiology/physiology ; Male ; Mosquito Control/*methods/statistics & numerical data ; Mosquito Vectors/growth & development/*microbiology/*physiology ; Population Dynamics ; Sex Characteristics ; Wolbachia/*physiology ; }, abstract = {The range of the mosquito Aedes aegypti continues to expand, putting more than two billion people at risk of arboviral infection. The sterile insect technique (SIT) has been used to successfully combat agricultural pests at large scale, but not mosquitoes, mainly because of challenges with consistent production and distribution of high-quality male mosquitoes. We describe automated processes to rear and release millions of competitive, sterile male Wolbachia-infected mosquitoes, and use of these males in a large-scale suppression trial in Fresno County, California. In 2018, we released 14.4 million males across three replicate neighborhoods encompassing 293 hectares. At peak mosquito season, the number of female mosquitoes was 95.5% lower (95% CI, 93.6-96.9) in release areas compared to non-release areas, with the most geographically isolated neighborhood reaching a 99% reduction. This work demonstrates the high efficacy of mosquito SIT in an area ninefold larger than in previous similar trials, supporting the potential of this approach in public health and nuisance-mosquito eradication programs.}, } @article {pmid32253286, year = {2020}, author = {Singh, R and Linksvayer, TA}, title = {Wolbachia-infected ant colonies have increased reproductive investment and an accelerated life cycle.}, journal = {The Journal of experimental biology}, volume = {223}, number = {Pt 9}, pages = {}, doi = {10.1242/jeb.220079}, pmid = {32253286}, issn = {1477-9145}, mesh = {Animals ; *Ants ; Life Cycle Stages ; Reproduction ; Sex Ratio ; *Wolbachia ; }, abstract = {Wolbachia is a widespread genus of maternally transmitted endosymbiotic bacteria that often manipulates the reproductive strategy and life history of its hosts to favor its own transmission. Wolbachia-mediated phenotypic effects are well characterized in solitary hosts, but effects in social hosts are unclear. The invasive pharaoh ant, Monomorium pharaonis, shows natural variation in Wolbachia infection between colonies and can be readily bred under laboratory conditions. We previously showed that Wolbachia-infected pharaoh ant colonies had more queen-biased sex ratios than uninfected colonies, which is expected to favor the spread of maternally transmitted Wolbachia Here, we further characterize the effects of Wolbachia on the short- and longer-term reproductive and life history traits of pharaoh ant colonies. First, we characterized the reproductive differences between naturally infected and uninfected colonies at three discrete time points and found that infected colonies had higher reproductive investment (i.e. infected colonies produced more new queens), particularly when existing colony queens were 3 months old. Next, we compared the long-term growth and reproduction dynamics of infected and uninfected colonies across their whole life cycle. Infected colonies had increased colony-level growth and early colony reproduction, resulting in a shorter colony life cycle, when compared with uninfected colonies.}, } @article {pmid32243448, year = {2020}, author = {Ross, PA and Axford, JK and Callahan, AG and Richardson, KM and Hoffmann, AA}, title = {Persistent deleterious effects of a deleterious Wolbachia infection.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {4}, pages = {e0008204}, pmid = {32243448}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Dengue/prevention & control ; Drosophila melanogaster ; *Evolution, Molecular ; Female ; Host Microbial Interactions ; Linear Models ; Male ; Mosquito Control ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods ; Population Dynamics ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Wolbachia are being used to reduce dengue transmission by Aedes aegypti mosquitoes around the world. To date releases have mostly involved Wolbachia strains with limited fitness effects but strains with larger fitness costs could be used to suppress mosquito populations. However, such infections are expected to evolve towards decreased deleterious effects. Here we investigate potential evolutionary changes in the wMelPop infection transferred from Drosophila melanogaster to Aedes aegypti more than ten years (~120 generations) ago. We show that most deleterious effects of this infection have persisted despite strong selection to ameliorate them. The wMelPop-PGYP infection is difficult to maintain in laboratory colonies, likely due to the persistent deleterious effects coupled with occasional maternal transmission leakage. Furthermore, female mosquitoes can be scored incorrectly as infected due to transmission of Wolbachia through mating. Infection loss in colonies was not associated with evolutionary changes in the nuclear background. These findings suggest that Wolbachia transinfections with deleterious effects may have stable phenotypes which could ensure their long-term effectiveness if released in natural populations to reduce population size.}, } @article {pmid32211166, year = {2020}, author = {Paredes-Montero, JR and Zia-Ur-Rehman, M and Hameed, U and Haider, MS and Herrmann, HW and Brown, JK}, title = {Genetic variability, community structure, and horizontal transfer of endosymbionts among three Asia II-Bemisia tabaci mitotypes in Pakistan.}, journal = {Ecology and evolution}, volume = {10}, number = {6}, pages = {2928-2943}, pmid = {32211166}, issn = {2045-7758}, abstract = {Endosymbionts associated with the whitefly Bemisia tabaci cryptic species are known to contribute to host fitness and environmental adaptation. The genetic diversity and population complexity were investigated for endosymbiont communities of B. tabaci occupying different micro-environments in Pakistan. Mitotypes of B. tabaci were identified by comparative sequence analysis of the mitochondria cytochrome oxidase I (mtCOI) gene sequence. Whitefly mitotypes belonged to the Asia II-1, -5, and -7 mitotypes of the Asia II major clade. The whitefly-endosymbiont communities were characterized based on 16S ribosomal RNA operational taxonomic unit (OTU) assignments, resulting in 43 OTUs. Most of the OTUs occurred in the Asia II-1 and II-7 mitotypes (r [2] = .9, p < .005), while the Asia II-5 microbiome was less complex. The microbiome OTU groups were mitotype-specific, clustering with a basis in phylogeographical distribution and the corresponding ecological niche of their whitefly host, suggesting mitotype-microbiome co-adaptation. The primary endosymbiont Portiera was represented by a single, highly homologous OTU (0%-0.67% divergence). Two of six Arsenophonus OTUs were uniquely associated with Asia II-5 and -7, and one occurred exclusively in Asia II-1, two only in Asia II-5, and one in both Asia II-1 and -7. Four other secondary endosymbionts, Cardinium, Hemipteriphilus, Rickettsia, and Wolbachia OTUs, were found at ≤29% frequencies. The most prevalent Arsenophonus OTU was found in all three Asia II mitotypes (55% frequency), whereas the same strain of Cardinium and Wolbachia was found in both Asia II-1 and -5, and a single Hemipteriphilus OTU occurred in Asia II-1 and -7. This pattern is indicative of horizontal transfer, suggestive of a proximity between mitotypes sufficient for gene flow at overlapping mitotype ecological niches.}, } @article {pmid32207826, year = {2020}, author = {Guz, N and Arshad, M and Cagatay, NS and Dageri, A and Ullah, MI}, title = {Detection of Wolbachia (Rickettsiales: Anaplasmataceae) and Candidatus Liberibacter asiaticus (Rhizobiales: Rhizobiaceae) Associated With Diaphorina citri (Hemiptera: Liviidae) Collected From Citrus reticulata (Sapindales: Rutaceae) and Alternate Host, Cordia myxa (Boraginales: Boraginaceae).}, journal = {Journal of economic entomology}, volume = {113}, number = {3}, pages = {1486-1492}, doi = {10.1093/jee/toaa043}, pmid = {32207826}, issn = {1938-291X}, mesh = {*Anaplasmataceae ; Animals ; *Boraginaceae ; *Citrus ; *Cordia ; *Hemiptera ; Plant Diseases ; *Rhizobiaceae ; Rickettsiales ; *Rutaceae ; *Wolbachia ; }, abstract = {The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an important insect pest of the citrus crop worldwide. It vectors the pathogen 'Candidatus Liberibacter asiaticus' (CLas) that causes a serious disease known as citrus greening. Here, we tested the infection frequency of Wolbachia and CLas from 100 D. citri individuals collected from two host plants belonging to families Rutaceae (Citrus reticulata Blanco) and Boraginaceae (Cordia myxa L.) using molecular methods. The following trend of endosymbionts infection in adult D. citri was found; 85.4% (35/41) by Wolbachia, and 19.5% (8/41) by CLas collected from C. reticulata plants and 65.4% (17/26) by Wolbachia, and 15.4% (4/26) by CLas in case of C. myxa plant. However, 61.5% (8/13) nymphs collected from C. reticulata and 20.0% (4/20) collected from C. myxa plants were infected by Wolbachia, while no nymph was infected by CLas collected from either host plants. Findings from this work represent the first report of CLas presence in D. citri feeding on C. myxa plants. By studying the presence of CLas with other endosymbiotic bacteria, future basic and applied research to develop control strategies can be prioritized.}, } @article {pmid32198842, year = {2020}, author = {Bing, XL and Zhao, DS and Peng, CW and Huang, HJ and Hong, XY}, title = {Similarities and spatial variations of bacterial and fungal communities in field rice planthopper (Hemiptera: Delphacidae) populations.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {947-963}, doi = {10.1111/1744-7917.12782}, pmid = {32198842}, issn = {1744-7917}, support = {804015//Nanjing Agricultural University/ ; 80900224//Nanjing Agricultural University/ ; 31672035//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Bacteria/classification/*isolation & purification ; China ; Environment ; Female ; Fungi/classification/*isolation & purification ; Hemiptera/*microbiology ; Male ; *Microbiota ; Mycobiome ; Phylogeny ; RNA, Bacterial/analysis ; RNA, Fungal/analysis ; RNA, Ribosomal, 16S/analysis ; Sex Factors ; Species Specificity ; Tissue Distribution ; }, abstract = {Rice planthoppers are notorious plant sap-feeding pests which cause serious damage. While several microbes in rice planthoppers have been broadly characterized, the abundance and diversity of bacteria and fungi in field planthoppers are largely unknown. This study investigated the bacterial and fungal community compositions of Chinese wild rice planthoppers Laodelphax striatellus and Sogatella furcifera using parallel 16S rRNA gene amplicon and internal transcribed space region sequencing. The bacteria varied significantly between the species and were partitioned significantly by sex, tissues and host environments in each species. The majority of bacteria were affiliated with the genera Wolbachia, Cardinium, Rickettsia and Pantoea. The abundance of Wolbachia was negatively correlated with that of Cardinium in both planthopper species. Compared with bacteria, the abundance and diversity of fungi did not differ between sexes but both were enriched in the gut. The bacterial community as a whole showed no significant correlation with the fungal community. The majority of fungi were related to Sarocladium, Alternaria, Malassezia, Aspergillus and Curvularia. A phylogenetic analysis revealed that these fungi were closely related to botanic symbionts or pathogens. Our results provide novel insights into the bacteria and fungi of rice planthoppers.}, } @article {pmid32191724, year = {2020}, author = {Dionysopoulou, NK and Papanastasiou, SA and Kyritsis, GA and Papadopoulos, NT}, title = {Effect of host fruit, temperature and Wolbachia infection on survival and development of Ceratitis capitata immature stages.}, journal = {PloS one}, volume = {15}, number = {3}, pages = {e0229727}, pmid = {32191724}, issn = {1932-6203}, mesh = {Animals ; Ceratitis capitata/*growth & development/*microbiology ; Fruit/*parasitology ; Larva/growth & development ; *Life Cycle Stages ; Proportional Hazards Models ; Pupa/growth & development ; Survival Analysis ; *Temperature ; Wolbachia/*physiology ; }, abstract = {The Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), holds an impressive record of successful invasions promoted by the growth and development of international fruit trade. Hence, survival of immatures within infested fruit that are subjected to various conditions during transportation seems to be a crucial feature that promotes invasion success. Wolbachia pipientis is a common endosymbiont of insects and other arthropods generating several biological effects on its hosts. Existing information report the influence of Wolbachia on the fitness traits of insect host species, including the Mediterranean fruit fly. However, little is known regarding effects of Wolbachia infection on immature development in different host fruits and temperatures. This study was conducted to determine the development and survival of immature stages of four different Mediterranean fruit fly populations, either infected or uninfected with Wolbachia, in two hosts (apples, bitter oranges) under three constant temperatures (15, 25 and 30°C), constant relative humidity (45-55 ± 5%), and a photoperiod of 14L:10D. Our findings demonstrate both differential response of two fruit fly lines to Wolbachia infection and differential effects of the two Wolbachia strains on the same Mediterranean fruit fly line. Larva-to-pupa and larva-to-adult survival followed similar patterns and varied a lot among the four medfly populations, the two host fruits and the different temperatures. Pupation rates and larval developmental time were higher for larvae implanted in apples compared to bitter oranges. The survival rates of wildish medflies were higher than those of the laboratory adapted ones, particularly in bitter oranges. The Wolbachia infected medflies, expressed lower survival rates and higher developmental times, especially the wCer4 infected line. High temperatures constrained immature development and were lethal for the Wolbachia infected wCer4 medfly line. Lower temperatures inferred longer developmental times to immature stages of all medfly populations tested, in both host fruits. Implications on the ecology and survival of the fly in nature are discussed.}, } @article {pmid32189066, year = {2020}, author = {Fallon, AM}, title = {Computational evidence for antitoxins associated with RelE/ParE, RatA, Fic, and AbiEii-family toxins in Wolbachia genomes.}, journal = {Molecular genetics and genomics : MGG}, volume = {295}, number = {4}, pages = {891-909}, doi = {10.1007/s00438-020-01662-0}, pmid = {32189066}, issn = {1617-4623}, mesh = {Animals ; Chromosomes, Bacterial/genetics ; Culicidae/microbiology ; DNA Topoisomerase IV/genetics ; Genome, Bacterial/genetics ; Humans ; Male ; Parthenogenesis/*genetics ; Pest Control ; Reproduction/*genetics ; Symbiosis/genetics ; Toxin-Antitoxin Systems/*genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia is an obligate intracellular Gram-negative alpha-proteobacterium that has diverse effects on reproduction of arthropod hosts, including cytoplasmic incompatibility, male killing, feminization, and parthenogenesis. Some of these effects have important potential for control of insect pests, including mosquitoes that vector pathogens of humans. In mosquitoes, and in most other arthropods, elimination of Wolbachia by antibiotic treatment has no effect on host survival and reverses the Wolbachia-associated phenotype. Elimination of Wolbachia strain wFol, which enables parthenogenetic reproduction of the Collembolan, Folsomia candida, would result in population extinction. However, F. candida adults remain viable and resume reproduction when antibiotics are removed, suggesting that wFol survives antibiotic treatment in a quiescent persister state similar to that induced by chromosomally encoded toxin-antitoxin (TA) modules in free-living bacteria. Computational approaches were used to document the presence of antitoxin genes upstream of Wolbachia RelE/ParE, Fic, and AbiEii toxin genes. Moreover, this analysis revealed that Wolbachia RatA toxin is encoded by a single copy gene associated with an ssrS noncoding RNA gene. Documentation of potentially functional TA modules expands our understanding of the metabolic capabilities of Wolbachia, and provides an explanation for variable and sometimes contradictory results of antibiotic treatments. The presence of chromosomal TA modules in Wolbachia genomes suggests that wFol, and potentially other strains of Wolbachia, can enter a quiescent persister state.}, } @article {pmid32187233, year = {2020}, author = {Wu, X and Lindsey, ARI and Chatterjee, P and Werren, JH and Stouthamer, R and Yi, SV}, title = {Distinct epigenomic and transcriptomic modifications associated with Wolbachia-mediated asexuality.}, journal = {PLoS pathogens}, volume = {16}, number = {3}, pages = {e1008397}, pmid = {32187233}, issn = {1553-7374}, mesh = {Animals ; *DNA Methylation ; *DNA, Protozoan/genetics/metabolism ; Epigenome/*physiology ; *Gene Expression Regulation ; Genome-Wide Association Study ; Transcriptome/*physiology ; Wasps/parasitology ; *Wolbachia/genetics/metabolism ; }, abstract = {Wolbachia are maternally transmitted intracellular bacteria that induce a range of pathogenic and fitness-altering effects on insect and nematode hosts. In parasitoid wasps of the genus Trichogramma, Wolbachia infection induces asexual production of females, thus increasing transmission of Wolbachia. It has been hypothesized that Wolbachia infection accompanies a modification of the host epigenome. However, to date, data on genome-wide epigenomic changes associated with Wolbachia are limited, and are often confounded by background genetic differences. Here, we took sexually reproducing Trichogramma free of Wolbachia and introgressed their genome into a Wolbachia-infected cytoplasm, converting them to Wolbachia-mediated asexuality. Wolbachia was then cured from replicates of these introgressed lines, allowing us to examine the genome-wide effects of wasps newly converted to asexual reproduction while controlling for genetic background. We thus identified gene expression and DNA methylation changes associated with Wolbachia-infection. We found no overlaps between differentially expressed genes and differentially methylated genes, indicating that Wolbachia-infection associated DNA methylation change does not directly modulate levels of gene expression. Furthermore, genes affected by these mechanisms exhibit distinct evolutionary histories. Genes differentially methylated due to the infection tended to be evolutionarily conserved. In contrast, differentially expressed genes were significantly more likely to be unique to the Trichogramma lineage, suggesting host-specific transcriptomic responses to infection. Nevertheless, we identified several novel aspects of Wolbachia-associated DNA methylation changes. Differentially methylated genes included those involved in oocyte development and chromosome segregation. Interestingly, Wolbachia-infection was associated with higher levels of DNA methylation. Additionally, Wolbachia infection reduced overall variability in gene expression, even after accounting for the effect of DNA methylation. We also identified specific cases where alternative exon usage was associated with DNA methylation changes due to Wolbachia infection. These results begin to reveal distinct genes and molecular pathways subject to Wolbachia induced epigenetic modification and/or host responses to Wolbachia-infection.}, } @article {pmid32174906, year = {2020}, author = {van den Bosch, TJM and Welte, CU}, title = {The Microbial Diversity of Cabbage Pest Delia radicum Across Multiple Life Stages.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {315}, pmid = {32174906}, issn = {1664-302X}, abstract = {The cabbage root fly Delia radicum is a worldwide pest that causes yield losses of many common cabbage crops. The bacteria associated with D. radicum are suggested to influence the pest status of their host. In this study, we characterized insect-associated bacteria of D. radicum across multiple life stages and of their diet plant (turnip, Brassica rapa subsp. rapa) by sequencing the V3-V4 region of 16S rRNA genes using the Illumina MiSeq platform. In total, over 1.2M paired-end reads were obtained, identifying 1006 bacterial amplicon sequence variants (ASVs) in samples obtained from the eggs, larvae, pupae and adults of D. radicum, as well as turnips that were either fresh or infested with D. radicum larvae. The microbial community in D. radicum was dominated by Wolbachia, a common endosymbiont of arthropods which we found in all of the investigated insect samples, with the pupal stage having the highest relative abundance. Moderate amounts of Firmicutes were found only in adult D. radicum flies, but not in previous life stages. Actinobacteria were mostly found on the eggs and on the skin of fresh plants on which the eggs were deposited. These plants also harbored a large amount of Pseudomonas. The bacterial diversity of the healthy turnip was low, whereas the microbial community of decaying turnips that were heavily infested by D. radicum larvae and showing symptoms of advanced soft rot was characterized by a high bacterial diversity. Taken together, this work provides insights into the bacterial communities associated with the cabbage pest D. radicum and its associated disease symptoms.}, } @article {pmid32168905, year = {2020}, author = {Rossitto De Marchi, B and Smith, HA}, title = {Bacterial Endosymbiont Diversity Among Bemisia tabaci (Hemiptera: Aleyrodidae) Populations in Florida.}, journal = {Insects}, volume = {11}, number = {3}, pages = {}, pmid = {32168905}, issn = {2075-4450}, abstract = {The sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), is a pest of many economically important agricultural crops and a vector of plant viruses. Bemisia tabaci harbors facultative endosymbiont species that have been implicated in pest status, including tolerance to insecticides, virus transmission efficiency and tolerance to high-temperatures. The facultative endosymbionts reported in B. tabaci include Arsenophonus, Hamiltonella, Wolbachia, Cardinium, Fritschea and Rickettsia. We collected whitefly populations from weed and crop hosts in south Florida and identified the whitefly species as well as the facultative endosymbionts present in these populations by molecular analysis. In addition, a phylogenetic analysis of whiteflies and their endosymbionts was performed. The only facultative endosymbionts found among the B. tabaci populations collected in Florida were Hamiltonella and Rickettsia. The phylogenetic analysis revealed the low genetic diversity of whiteflies and their endosymbionts. Additionally, the phylogenetic tree clustered Rickettsia from Florida in the R1 genetic group. The results will aid to understand the role of the bacterial endosymbionts in the whitefly host.}, } @article {pmid32164779, year = {2020}, author = {Hu, Y and Xi, Z and Liu, X and Wang, J and Guo, Y and Ren, D and Wu, H and Wang, X and Chen, B and Liu, Q}, title = {Correction to: Identification and molecular characterization of Wolbachia strains in natural populations of Aedes albopictus in China.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {124}, pmid = {32164779}, issn = {1756-3305}, abstract = {Following publication of the original article [1], the corresponding author flagged that the article had published with two errors.}, } @article {pmid32163151, year = {2020}, author = {Morrow, JL and Schneider, DI and Klasson, L and Janitz, C and Miller, WJ and Riegler, M}, title = {Parallel Sequencing of Wolbachia wCer2 from Donor and Novel Hosts Reveals Multiple Incompatibility Factors and Genome Stability after Host Transfers.}, journal = {Genome biology and evolution}, volume = {12}, number = {5}, pages = {720-735}, pmid = {32163151}, issn = {1759-6653}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Bacterial Proteins/*genetics ; Cytoplasm/*genetics ; Drosophila/*microbiology ; Evolution, Molecular ; *Genomic Instability ; *Host Specificity ; Host-Pathogen Interactions ; Phenotype ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {The application of Wolbachia in insect pest and vector control requires the establishment of genotypically stable host associations. The cytoplasmic incompatibility (CI) inducing Wolbachia strain wCer2 naturally occurs in the cherry fruit fly Rhagoletis cerasi as co-infection with other strains and was transferred to other fruit fly species by embryonic microinjections. We obtained wCer2 genome data from its native and three novel hosts, Drosophila simulans, Drosophila melanogaster, and Ceratitis capitata and assessed its genome stability, characteristics, and CI factor (cif) genes. De novo assembly was successful from Wolbachia cell-enriched singly infected D. simulans embryos, with minimal host and other bacterial genome traces. The low yield of Wolbachia sequence reads from total genomic extracts of one multiply infected R. cerasi pupa and one singly infected C. capitata adult limited de novo assemblies but was sufficient for comparative analyses. Across hosts wCer2 was stable in genome synteny and content. Polymorphic nucleotide sites were found in wCer2 of each host; however, only one nucleotide was different between R. cerasi and C. capitata, and none between replicated D. simulans lines. The wCer2 genome is highly similar to wAu (D. simulans), wMel (D. melanogaster), and wRec (Drosophila recens). In contrast to wMel and wRec (each with one cif gene pair) and wAu (without any cif genes), wCer2 has three pairs of Type I cif genes, and one Type V cifB gene without a cifA complement. This may explain previously reported CI patterns of wCer2, including incomplete rescue of its own CI modification in three novel host species.}, } @article {pmid32155143, year = {2020}, author = {Mancini, MV and Herd, CS and Ant, TH and Murdochy, SM and Sinkins, SP}, title = {Wolbachia strain wAu efficiently blocks arbovirus transmission in Aedes albopictus.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {3}, pages = {e0007926}, pmid = {32155143}, issn = {1935-2735}, support = {MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 108508/Z/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; *Antibiosis ; Arbovirus Infections/prevention & control ; Arboviruses/*growth & development ; Disease Transmission, Infectious/prevention & control ; Drosophila/microbiology ; Female ; Male ; Mosquito Vectors/microbiology/virology ; Wolbachia/*growth & development/isolation & purification ; }, abstract = {The global incidence of arboviral diseases transmitted by Aedes mosquitoes, including dengue, chikungunya, yellow fever, and Zika, has increased dramatically in recent decades. The release of Aedes aegypti carrying the maternally inherited symbiont Wolbachia as an intervention to control arboviruses is being trialled in several countries. However, these efforts are compromised in many endemic regions due to the co-localization of the secondary vector Aedes albopictus, the Asian tiger mosquito. Ae. albopictus has an expanding global distribution following incursions into a number of new territories. To date, only the wMel and wPip strains of Wolbachia have been reported to be transferred into and characterized in this vector. A Wolbachia strain naturally infecting Drosophila simulans, wAu, was selected for transfer into a Malaysian Ae. albopictus line to create a novel triple-strain infection. The newly generated line showed self-compatibility, moderate fitness cost and complete resistance to Zika and dengue infections.}, } @article {pmid32144639, year = {2020}, author = {Duan, DY and Liu, GH and Cheng, TY}, title = {Microbiome analysis of the saliva and midgut from partially or fully engorged female adult Dermacentor silvarum ticks in China.}, journal = {Experimental & applied acarology}, volume = {80}, number = {4}, pages = {543-558}, pmid = {32144639}, issn = {1572-9702}, support = {No. 2018JJ2167//the Natural Science Foundation of Hunan Province, China/ ; No. SYL201802016//the Double First-Class Construction Project of Hunan Agricultural University/ ; No. 18QN24//the Youth Science Foundation of Hunan Agricultural University/ ; No. 31902294//the National Science Foundation for Young Scientists of China/ ; }, mesh = {Animals ; China ; Dermacentor/*microbiology ; Female ; Gastrointestinal Tract/*microbiology ; *Microbiota ; RNA, Ribosomal, 16S ; Saliva/*microbiology ; }, abstract = {Dermacentor silvarum is widely distributed in northern China and transmits several pathogens that cause diseases in humans and domestic animals. We analysed the comprehensive bacterial community of the saliva and midgut from partially and fully engorged female adult D. silvarum. Dermacentor silvarum samples were collected from Guyuan, China. Bacterial DNA was extracted from the saliva and midgut contents of partially or fully engorged female adult D. silvarum. Sequencing of the V3-V4 hypervariable regions of the 16S rRNA genes was performed using the IonS5[TM]XL platform. The bacterial diversity in saliva was higher than in the midgut. The bacterial diversity of saliva from fully engorged ticks was greater than in partially engorged tick saliva. The bacterial diversity in midguts from partially engorged ticks was greater than in fully engorged tick midguts. Proteobacteria was the most dominant bacterial phylum in all of the samples. Twenty-nine bacterial genera were detected in all of the samples. Rickettsia, Anaplasma, and Stenotrophomonas were the main genera. The symbionts Coxiella, Arsenophonus, and Wolbachia were also detected in all of the samples. Eight bacterial species were identified in all of the experimental samples. Anaplasma marginale was reported for the first time in D. silvarum.}, } @article {pmid32144396, year = {2020}, author = {Epis, S and Varotto-Boccazzi, I and Crotti, E and Damiani, C and Giovati, L and Mandrioli, M and Biggiogera, M and Gabrieli, P and Genchi, M and Polonelli, L and Daffonchio, D and Favia, G and Bandi, C}, title = {Chimeric symbionts expressing a Wolbachia protein stimulate mosquito immunity and inhibit filarial parasite development.}, journal = {Communications biology}, volume = {3}, number = {1}, pages = {105}, pmid = {32144396}, issn = {2399-3642}, mesh = {Acetobacteraceae/genetics/*metabolism ; Aedes/immunology/*microbiology ; Animals ; Anopheles/*microbiology/parasitology ; Bacterial Outer Membrane Proteins/genetics/*metabolism ; Dirofilaria immitis/growth & development/*microbiology ; Host-Parasite Interactions ; Membrane Proteins/genetics/*metabolism ; Phagocytosis ; Symbiosis ; Wolbachia/genetics/*metabolism ; }, abstract = {Wolbachia can reduce the capability of mosquitoes to transmit infectious diseases to humans and is currently exploited in campaigns for the control of arboviruses, like dengue and Zika. Under the assumption that Wolbachia-mediated activation of insect immunity plays a role in the reduction of mosquito vectorial capacity, we focused our attention on the Wolbachia surface protein (WSP), a potential inductor of innate immunity. We hypothesized that the heterologous expression of this protein in gut- and tissue-associated symbionts may reduce parasite transmission. We thus engineered the mosquito bacterial symbiont Asaia to express WSP (Asaia[WSP]). Asaia[WSP] induced activation of the host immune response in Aedes aegypti and Anopheles stephensi mosquitoes, and inhibited the development of the heartworm parasite Dirofilaria immitis in Ae. aegypti. These results consolidate previous evidence on the immune-stimulating property of WSP and make Asaia[WSP] worth of further investigations as a potential tool for the control of mosquito-borne diseases.}, } @article {pmid32142516, year = {2020}, author = {O'Driscoll, M and Imai, N and Ferguson, NM and Hadinegoro, SR and Satari, HI and Tam, CC and Dorigatti, I}, title = {Spatiotemporal variability in dengue transmission intensity in Jakarta, Indonesia.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {3}, pages = {e0008102}, pmid = {32142516}, issn = {1935-2735}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; U01 GM110721/GM/NIGMS NIH HHS/United States ; }, mesh = {Adolescent ; Adult ; Age Distribution ; Aged ; Child ; Child, Preschool ; Cities/epidemiology ; Dengue/*epidemiology/*transmission ; Disease Notification/statistics & numerical data ; *Disease Transmission, Infectious ; Female ; Humans ; Indonesia/epidemiology ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Models, Theoretical ; Spatio-Temporal Analysis ; Young Adult ; }, abstract = {BACKGROUND: Approximately 70% of the global burden of dengue disease occurs on the Asian continent, where many large urban centres provide optimal environments for sustained endemic transmission and periodic epidemic cycles. Jakarta, the capital of Indonesia, is a densely populated megacity with hyperendemic dengue transmission. Characterization of the spatiotemporal distribution of dengue transmission intensity is of key importance for optimal implementation of novel control and prevention programmes, including vaccination. In this paper we use mathematical models to provide the first detailed description of spatial and temporal variability in dengue transmission intensity in Jakarta.

We applied catalytic models in a Bayesian framework to age-stratified dengue case notification data to estimate dengue force of infection and reporting probabilities in 42 subdistricts of Jakarta. The model was fitted to yearly and average annual data covering a 10-year period between 2008 and 2017. We estimated a long-term average annual transmission intensity of 0.130 (95%CrI: 0.129-0.131) per year in Jakarta province, ranging from 0.090 (95%CrI: 0.077-0.103) to 0.164 (95%CrI: 0.153-0.174) across subdistricts. Annual average transmission intensity in Jakarta province during the 10-year period ranged from 0.012 (95%CrI: 0.011-0.013) in 2017 to 0.124 (95%CrI: 0.121-0.128) in 2016.

CONCLUSIONS/SIGNIFICANCE: While the absolute number of dengue case notifications cannot be relied upon as a measure of endemicity, the age-distribution of reported dengue cases provides valuable insights into the underlying nature of transmission. Our estimates from yearly and average annual case notification data represent the first detailed estimates of dengue transmission intensity in Jakarta's subdistricts. These will be important to consider when assessing the population-level impact and cost-effectiveness of potential control and prevention programmes in Jakarta province, such as the controlled release of Wolbachia-carrying mosquitoes and vaccination.}, } @article {pmid32141700, year = {2020}, author = {Glowska, E and Filutowska, ZK and Dabert, M and Gerth, M}, title = {Microbial composition of enigmatic bird parasites: Wolbachia and Spiroplasma are the most important bacterial associates of quill mites (Acariformes: Syringophilidae).}, journal = {MicrobiologyOpen}, volume = {9}, number = {5}, pages = {e964}, pmid = {32141700}, issn = {2045-8827}, mesh = {Animals ; Biodiversity ; Birds/*parasitology ; DNA, Bacterial/genetics ; Feathers/*parasitology ; Microbiota ; Mites/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Spiroplasma/*classification/isolation & purification ; Symbiosis ; Wolbachia/*classification/isolation & purification ; }, abstract = {BACKGROUND: The microbiome is an integral component of many animal species, potentially affecting behavior, physiology, and other biological properties. Despite this importance, bacterial communities remain vastly understudied in many groups of invertebrates, including mites. Quill mites (Acariformes: Syringophilidae) are a poorly known group of permanent bird ectoparasites that occupy quills of feathers and feed on bird subcutaneous tissue and fluids. Most of the known species have strongly female-biased sex ratio, and it was hypothesized that this is caused by endosymbiotic bacteria. Previously, Anaplasma phagocytophilum (Foggie) and a high diversity of Wolbachia strains were detected in quill mites via targeted PCR screens. Here, we use an unbiased 16S rRNA gene amplicon sequencing approach to determine other bacteria that potentially impact quill mite biology.

RESULTS: We performed 16S rRNA gene amplicon sequencing of 126 quill mite individuals from eleven species parasitizing twelve species (four families) of passeriform birds. In addition to Wolbachia, we found Spiroplasma as potential symbiont of quill mites. Consistently, high Spiroplasma titers were only found in individuals of two mite species associated with finches of the genus Carduelis, suggesting a history of horizontal transfers of Spiroplasma via the bird host. Furthermore, there was evidence for Spiroplasma negatively affecting Wolbachia titers. We found no evidence for the previously reported Anaplasma in quill mites, but detected sequences of high similarity to the potential pathogens Brucella and Bartonella at low abundances. Other amplicon sequence variants (ASVs) could be assigned to a diverse number of bacterial taxa, including several that were previously isolated from bird skin. Further, many frequently found ASVs were assigned to taxa that show a very broad distribution with no strong prior evidence for symbiotic association with animals. We interpret these findings as evidence for a scarcity of resident microbial associates (other than inherited symbionts) in quill mites.}, } @article {pmid32138145, year = {2020}, author = {Bubici, G and Prigigallo, MI and Garganese, F and Nugnes, F and Jansen, M and Porcelli, F}, title = {First Report of Aleurocanthus spiniferus on Ailanthus altissima: Profiling of the Insect Microbiome and MicroRNAs.}, journal = {Insects}, volume = {11}, number = {3}, pages = {}, pmid = {32138145}, issn = {2075-4450}, support = {CoBIAs/AGG2016//Apulia region, Italy/ ; }, abstract = {We report the first occurrence of the orange spiny whitefly (Aleurocanthus spiniferus; OSW) on the tree of heaven (Ailanthus altissima) in Bari, Apulia region, Italy. After our first observation in 2016, the infestation recurred regularly during the following years and expanded to the neighboring trees. Since then, we have also found the insect on numerous patches of the tree of heaven and other plant species in the Bari province. Nevertheless, the tree of heaven was not particularly threatened by the insect, so that a possible contribution by OSW for the control of such an invasive plant cannot be hypothesized hitherto. This work was also aimed at profiling the microbiome of OSW feeding on A. altissima. For this purpose, we used the denaturing gradient gel electrophoresis (DGGE) and the deep sequencing of small RNAs (sRNAs). Both techniques unveiled the presence of "Candidatus Portiera" (primary endosymbiont), Wolbachia sp. and Rickettsia sp., endosymbionts already reported for other Aleyrodidae. Deep sequencing data were analyzed by four computational pipelines in order to understand the reliability of the detection of fungi, bacteria, and viruses: Kraken, Kaiju, Velvet, and VelvetOptimiser. Some contigs assembled by Velvet or VelvetOptimiser were associated with insects, but not necessarily in the Aleurocanthus genus or Aleyrodidae family, suggesting the non-specificity of sRNAs or possible traces of parasitoids in the sample (e.g., Eretmocerus sp.). Finally, deep sequencing data were used to describe the microtranscriptome of OSW: 56 canonical and at least four high-confidence novel microRNAs (miRNAs) were identified. The overall miRNA abundance in OSW was in agreement with previous works on Bemisia tabaci, and bantam-3p, miR-276a-3p, miR-317-3p, miR-750-3p, and mir-8-3p were the most represented miRNAs.}, } @article {pmid32132880, year = {2020}, author = {Nikolouli, K and Sassù, F and Mouton, L and Stauffer, C and Bourtzis, K}, title = {Combining sterile and incompatible insect techniques for the population suppression of Drosophila suzukii.}, journal = {Journal of pest science}, volume = {93}, number = {2}, pages = {647-661}, pmid = {32132880}, issn = {1612-4758}, abstract = {The spotted wing Drosophila, Drosophila suzukii, has recently invaded Europe and the Americas, and it is a major threat for a wide variety of commercial soft fruits both in open field and greenhouse production systems. D. suzukii infests a wide range of ripening fruits, leading to substantial yield and revenue losses. As the application of insecticides close to the harvest period poses great concerns, the development of an efficient environment-friendly control approach to fight D. suzukii is necessary. In this study, we exploited the sterile insect technique (SIT) in combination with Wolbachia symbiosis as a population suppression approach that can constitute a potential component of an area-wide integrated pest management program. We aimed to establish a combined SIT/incompatible insect technique (IIT) protocol that would require lower irradiation doses as a complementary tool for D. suzukii management. Two D. suzukii lines trans-infected with the Wolbachia wHa and wTei strains were irradiated at doses four times less than usual (e.g., 45 Gy), and the egg hatching and adult emergence were determined. Our results indicated that wHa and wTei females as well as wHa males were sterile at this low dose. The longevity, adult emergence and flight ability of adults were evaluated, and no major effect caused by irradiation was detected. Our data indicate that a SIT/IIT protocol can be a competent approach for D. suzukii management.}, } @article {pmid32130504, year = {2020}, author = {Gallo-Franco, JJ and Toro-Perea, N}, title = {Variations in the Bacterial Communities in Anastrepha obliqua (Diptera: Tephritidae) According to the Insect Life Stage and Host Plant.}, journal = {Current microbiology}, volume = {77}, number = {7}, pages = {1283-1291}, doi = {10.1007/s00284-020-01939-y}, pmid = {32130504}, issn = {1432-0991}, mesh = {Animals ; Bacteria/classification/genetics ; Fruit/microbiology ; Larva/*microbiology ; Magnoliopsida/*microbiology ; Metagenome/*genetics ; Microbiota/*genetics ; Tephritidae/*microbiology ; }, abstract = {Insects have established close relationships with a wide variety of microorganisms, which play a key role in insect ecology and evolution. Fruit flies in the Tephritidae family have economic importance at the global level, including species such as Anastrepha obliqua, which is an important pest in the neotropical region. Although several studies have been performed on the microbiota associated with fruit flies, there are still large gaps in our knowledge about the bacterial communities on the genus Anastrepha. During this study, we used high-throughput sequencing to characterize the bacterial communities of the polyphagous fly A. obliqua, and we evaluated the effect of the life stage (larvae and adults) and host plant (three plant species) on the structure of these communities. Our results show that the bacterial communities in A. obliqua appears to be structured according to the insect life stage and the host plant. The predominant genera belonging to the phylum Proteobacteria were Wolbachia and Enterobacter in both larvae and adults, and they displayed differences in abundance between them, with Wolbachia sp. being more abundant in larvae and Enterobacter sp. being more abundant in adults. Differences in the structures of the bacterial communities were also observed according to the host plant with higher abundance of Enterobacter and Acetobacter bacteria in mango and plum fruits. Based on our results, it can be hypothesized that the bacterial communities on A. obliqua reorganize according to the needs of these insects during their different life stages and could also play an important role in the establishment of this fly species on different host plants. This study represents the first approach to understanding microorganism-insect interactions in fruit flies in Colombia.}, } @article {pmid32128956, year = {2020}, author = {Li, Y and Liu, X and Wang, N and Zhang, Y and Hoffmann, AA and Guo, H}, title = {Background-dependent Wolbachia-mediated insecticide resistance in Laodelphax striatellus.}, journal = {Environmental microbiology}, volume = {22}, number = {7}, pages = {2653-2663}, doi = {10.1111/1462-2920.14974}, pmid = {32128956}, issn = {1462-2920}, support = {31672027//the National Natural Science Foundation of China/International ; 31972265//the National Natural Science Foundation of China/International ; //a Fellowship from the National Health and Medical Research Council/International ; cx(16)1001//the Independent Innovation Fund of Agricultural Science and Technology in Jiangsu province, China/International ; }, mesh = {Animals ; China ; Hemiptera/*drug effects/microbiology ; Insecticide Resistance/*physiology ; Insecticides/*pharmacology ; Thiadiazines/*pharmacology ; Wolbachia/*metabolism ; }, abstract = {Although facultative endosymbionts are now known to protect insect hosts against pathogens and parasitoids, the effects of endosymbionts on insecticide resistance are still unclear. Here we show that Wolbachia are associated with increased resistance to the commonly used insecticide, buprofezin, in the small brown planthopper (Laodelphax striatellus) in some genetic backgrounds while having no effect in other backgrounds. In three Wolbachia-infected lines from experimental buprofezin-resistant strains and one line from a buprofezin-susceptible line established from Chuxiong, Yunnan province, China, susceptibility to buprofezin increased after removal of Wolbachia. An increase in susceptibility was also evident in a Wolbachia-infected line established from a field population in Rugao, Jiangsu province. However, no increase was evident in two field populations from Nanjing and Fengxian, Jiangsu province, China. When Wolbachia was introgressed into different genetic backgrounds, followed by Wolbachia removal, the data pointed to Wolbachia effects that depend on the nuclear background as well as on the Wolbachia strain. However, there was no relationship between Wolbachia density and the component of buprofezin resistance associated with the symbiont. The results suggest that Wolbachia effects associated with chemical resistance are complex and unpredictable, but also that they can be substantial.}, } @article {pmid32126342, year = {2020}, author = {Turner, JD and Marriott, AE and Hong, D and O' Neill, P and Ward, SA and Taylor, MJ}, title = {Novel anti-Wolbachia drugs, a new approach in the treatment and prevention of veterinary filariasis?.}, journal = {Veterinary parasitology}, volume = {279}, number = {}, pages = {109057}, doi = {10.1016/j.vetpar.2020.109057}, pmid = {32126342}, issn = {1873-2550}, support = {MC_PC_17167/MRC_/Medical Research Council/United Kingdom ; MC_PC_18055/MRC_/Medical Research Council/United Kingdom ; NC/S001131/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; }, mesh = {Animals ; *Anti-Bacterial Agents ; Dirofilaria immitis/*drug effects ; Dirofilaria repens/*drug effects ; Dirofilariasis/*prevention & control ; Doxycycline/*pharmacology ; Filaricides/*pharmacology ; Wolbachia ; }, abstract = {Filarial nematodes are tissue-dwelling parasitic worms that can cause a range of disfiguring pathologies in humans and potentially lethal infections of companion animals. The bacterial endosymbiont, Wolbachia, is present within most human and veterinary filarial pathogens, including the causative agent of heartworm disease, Dirofilaria immitis. Doxycycline-mediated drug targeting of Wolbachia leads to sterility, clearance of microfilariae and gradual death of adult filariae. This mode of action is attractive in the treatment of filariasis because it avoids severe host inflammatory adverse reactions invoked by rapid-killing anthelmintic agents. However, doxycycline needs to be taken for four weeks to exert curative activity. In this review, we discuss the evidence that Wolbachia drug targeting is efficacious in blocking filarial larval development as well as in the treatment of chronic filarial disease. We present the current portfolio of next-generation anti-Wolbachia candidates discovered through phenotypic screening of chemical libraries and validated in a range of in vitro and in vivo filarial infection models. Several novel chemotypes have been identified with selected narrow-spectrum anti-Wolbachia specificity and superior time-to-kill kinetics compared with doxycycline. We discuss the opportunities of developing these novel anti-Wolbachia agents as either cures, adjunct therapies or new preventatives for the treatment of veterinary filariasis.}, } @article {pmid32123899, year = {2020}, author = {Ourry, M and Lopez, V and Hervé, M and Lebreton, L and Mougel, C and Outreman, Y and Poinsot, D and Cortesero, AM}, title = {Long-lasting effects of antibiotics on bacterial communities of adult flies.}, journal = {FEMS microbiology ecology}, volume = {96}, number = {4}, pages = {}, doi = {10.1093/femsec/fiaa028}, pmid = {32123899}, issn = {1574-6941}, mesh = {Animals ; *Anti-Bacterial Agents/pharmacology ; Bacteria/genetics ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Tetracycline/pharmacology ; }, abstract = {Insect symbionts benefit their host and their study requires large spectrum antibiotic use like tetracycline to weaken or suppress symbiotic communities. While antibiotics have a negative impact on insect fitness, little is known about antibiotic effects on insect microbial communities and how long they last. We characterized the bacterial communities of adult cabbage root fly Delia radicum in a Wolbachia-free population and evaluated the effect of tetracycline treatment on these communities over several generations. Three D. radicum generations were used: the first- and second-generation flies either ingested tetracycline or not, while the third-generation flies were untreated but differed with their parents and/or grandparents that had or had not been treated. Fly bacterial communities were sequenced using a 16S rRNA gene. Tetracycline decreased fly bacterial diversity and induced modifications in both bacterial abundance and relative frequencies, still visible on untreated offspring whose parents and/or grandparents had been treated, therefore demonstrating long-lasting transgenerational effects on animal microbiomes after antibiotic treatment. Flies with an antibiotic history shared bacterial genera, potentially tetracycline resistant and heritable. Next, the transmission should be investigated by comparing several insect development stages and plant compartments to assess vertical and horizontal transmissions of D. radicum bacterial communities.}, } @article {pmid32122896, year = {2020}, author = {Pietri, JE and Liang, D}, title = {Insecticidal Activity of Doxycycline against the Common Bedbug.}, journal = {Antimicrobial agents and chemotherapy}, volume = {64}, number = {5}, pages = {}, pmid = {32122896}, issn = {1098-6596}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Bedbugs/*drug effects/*microbiology ; Doxycycline/*pharmacology ; Insecticides/pharmacology ; Penicillins/pharmacology ; Rifampin/pharmacology ; Symbiosis/drug effects ; Wolbachia/*drug effects/metabolism ; }, abstract = {There is an ongoing need for safe and effective anti-bedbug compounds. Here, we tested the toxicity of three antimicrobial agents against bedbugs when administered orally. We reveal that doxycycline has direct insecticidal activity at 250 μg/ml (0.025%) that is particularly strong against immature bedbugs and appears to be independent of antimicrobial activity. Future studies to determine the mechanisms behind this property could be useful for the development of orally active insecticides or anti-bedbug therapeutics.}, } @article {pmid32112519, year = {2020}, author = {Zhou, JC and Liu, QQ and Wang, QR and Ning, SF and Che, WN and Dong, H}, title = {Optimal clutch size for quality control of bisexual and Wolbachia-infected thelytokous lines of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) mass reared on eggs of a substitutive host, Antheraea pernyi Guérin-Méneville (Lepidoptera: Saturniidae).}, journal = {Pest management science}, volume = {76}, number = {8}, pages = {2635-2644}, doi = {10.1002/ps.5805}, pmid = {32112519}, issn = {1526-4998}, support = {2017YFD0201805//National Key Research and Development Program of China/ ; 2016YFD0300704//National Key Research and Development Program of China/ ; 2017YFD0201000//National Key Research and Development Program of China/ ; }, mesh = {Animals ; China ; Clutch Size ; Female ; *Hymenoptera ; Male ; *Moths ; Ovum ; Quality Control ; *Wolbachia ; }, abstract = {BACKGROUND: Trichogramma dendrolimi has been widely used in augmentative biocontrol of lepidopteran pests in China. In mass production of T. dendrolimi using Antheraea pernyi eggs as substitutive hosts, which are large in size, as clutch size is a parameter of importance to produce high quality parasitoids. Here, we aimed to determine the optimal clutch size for the bisexual Wolbachia-uninfected line (TdB) and Wolbachia-infected thelytokous line (TdT) of T. dendrolimi reared on A. pernyi eggs.

RESULTS: A medium clutch size of 42.75 to 62.27 for TdB and 52.93 to 57.14 for TdT was optimal to maximize fitness-correlated traits of parasitoid individual. The optimal clutch sizes with maximized parameters included adult emergence rate, adult body size, adult longevity, fecundity, and sum of fecundity of all females per brood were 58.31 (86.00%), 42.75 (231.11 μm), 50.92 (2.69 days), 62.27 (150.89 eggs), and 83.25 (7926.33 eggs) for TdB and 57.14 (94.54%), 52.93 (236.97 μm), 53.64 (2.62 days), 56.80 (161.01 eggs), and 70.10 (8579.71 eggs) for TdT. The TdT had a shorter adult longevity, longer development time, and higher adult emergence rate than did its non-infected bisexual counterpart.

CONCLUSION: A medium brood size in a A. pernyi egg host was optimal to produce offspring parasitoids with higher fitness parameters for both bisexual Wolbachia-uninfected and thelytokous Wolbachia-infected lines of T. dendrolimi. The determination of optimal clutch size for T. dendrolimi will provide the reference for the quality control of T. dendrolimi production and improvement of the field performance of the wasps. © 2020 Society of Chemical Industry.}, } @article {pmid32092972, year = {2020}, author = {Kaczmarczyk-Ziemba, A and Zagaja, M and Wagner, GK and Pietrykowska-Tudruj, E and Staniec, B}, title = {First Insight into Microbiome Profiles of Myrmecophilous Beetles and Their Host, Red Wood Ant Formica polyctena (Hymenoptera: Formicidae)-A Case Study.}, journal = {Insects}, volume = {11}, number = {2}, pages = {}, pmid = {32092972}, issn = {2075-4450}, support = {EZ.0290.1.28.2017//State Forests National Forest Holding/ ; NB 520-3/2017//Polesie National Park/ ; }, abstract = {Formica polyctena belongs to the red wood ant species group. Its nests provide a stable, food rich, and temperature and humidity controlled environment, utilized by a wide range of species, called myrmecophiles. Here, we used the high-throughput sequencing of the 16S rRNA gene on the Illumina platform for identification of the microbiome profiles of six selected myrmecophilous beetles (Dendrophilus pygmaeus, Leptacinus formicetorum, Monotoma angusticollis, Myrmechixenus subterraneus, Ptenidium formicetorum and Thiasophila angulata) and their host F. polyctena. Analyzed bacterial communities consisted of a total of 23 phyla, among which Proteobacteria, Actinobacteria, and Firmicutes were the most abundant. Two known endosymbionts-Wolbachia and Rickettsia-were found in the analyzed microbiome profiles and Wolbachia was dominant in bacterial communities associated with F. polyctena, M. subterraneus, L. formicetorum and P. formicetorum (>90% of reads). In turn, M. angusticollis was co-infected with both Wolbachia and Rickettsia, while in the microbiome of T. angulata, the dominance of Rickettsia has been observed. The relationships among the microbiome profiles were complex, and no relative abundance pattern common to all myrmecophilous beetles tested was observed. However, some subtle, species-specific patterns have been observed for bacterial communities associated with D. pygmaeus, M. angusticollis, and T. angulata.}, } @article {pmid32091199, year = {2020}, author = {Hawryluk, NA}, title = {Macrofilaricides: An Unmet Medical Need for Filarial Diseases.}, journal = {ACS infectious diseases}, volume = {6}, number = {4}, pages = {662-671}, doi = {10.1021/acsinfecdis.9b00469}, pmid = {32091199}, issn = {2373-8227}, mesh = {Africa South of the Sahara ; Animals ; Anti-Bacterial Agents/administration & dosage ; Filaricides/*chemistry/*pharmacology ; Helminthiasis/classification/*drug therapy/parasitology ; Humans ; India ; Life Cycle Stages/drug effects ; Neglected Diseases/drug therapy/parasitology ; Wolbachia/drug effects/physiology ; }, abstract = {Neglected parasitic helminth diseases such as onchocerciasis and lymphatic filariasis affect an estimated 145 million people worldwide, creating a serious health burden in endemic areas such as sub-Saharan Africa and India. Although these diseases are not usually lethal, these filarial nematodes, transmitted by blood-feeding insect vectors, cause severe debilitation and cause chronic disability to infected individuals. The adult worms can reproduce from 5 to up to 14 years, releasing millions of microfilariae, juvenile worms, over an infected individual's lifetime. The current treatments for controlling human filarial infections is focused on killing microfilariae, the earliest larval stage. Currently, there is an unmet medical need for treatments consisting of a macrofilaricidal regimen, one that targets the adult stage of the parasite, to increase the rate of elimination, allow for safe use in coendemic regions of Onchocerca volvulus and Loa loa, and to provide a rapid method to resolve reinfections. Herein, recent approaches for targeting human filarial diseases are discussed, including direct acting agents to target parasitic nematodes and antibacterial approaches to target the endosymbiotic bacteria, Wolbachia.}, } @article {pmid32078985, year = {2020}, author = {Liberti, J and Engel, P}, title = {The gut microbiota - brain axis of insects.}, journal = {Current opinion in insect science}, volume = {39}, number = {}, pages = {6-13}, doi = {10.1016/j.cois.2020.01.004}, pmid = {32078985}, issn = {2214-5753}, mesh = {Animals ; Bees/*microbiology ; Behavior ; *Brain/microbiology/physiology ; Disease Models, Animal ; Drosophila/microbiology ; *Gastrointestinal Microbiome ; Host Microbial Interactions ; Hypocreales/pathogenicity ; Insecta/*microbiology ; Mental Disorders/microbiology ; Neurophysiology ; Symbiosis ; Wolbachia/pathogenicity ; }, abstract = {Research on the connections between gut microbes and the neurophysiology and behavior of their animal hosts has grown exponentially in just a few years. Most studies have focused on mammalian models as their relevance to human health is widely established. However, evidence is accumulating that insect behavior may be governed by molecular mechanisms that are partly homologous to those of mammals, and therefore relevant for the understanding of their behavioral dysfunctions. Social insects in particular may provide experimentally amenable models to disentangle the contributions of individual bacterial symbionts to the gut microbiota - brain axis. In this review, we summarize findings from recent research on the neurological and behavioral effects of the gut microbiota of insects and propose an integrated approach to unravel the extended behavioral phenotypes of gut microbes in the honey bee.}, } @article {pmid32078642, year = {2020}, author = {Zhang, D and Wang, Y and He, K and Yang, Q and Gong, M and Ji, M and Chen, L}, title = {Wolbachia limits pathogen infections through induction of host innate immune responses.}, journal = {PloS one}, volume = {15}, number = {2}, pages = {e0226736}, pmid = {32078642}, issn = {1932-6203}, mesh = {Animals ; *Culex/immunology/microbiology ; Immunity, Innate ; *Mosquito Vectors/immunology/microbiology ; Symbiosis ; Wolbachia/*immunology ; }, abstract = {BACKGROUND: Wolbachia has been reported to suppress a variety of pathogen infections in mosquitoes, but the mechanism is undefined. Two possibilities have been proposed. One is that Wolbachia activates host immune responses, and the other one is that Wolbachia competes with pathogens for limited nutrients.

In this study, we compared host immune responses and the densities of two different strains of Wolbachia in naturally occurring parental and artificially created hybrid host genetic backgrounds. No significant difference in Wolbachia density was found between these hosts. We found that Wolbachia could activate host innate immune responses when the host genetic profile was different from that of its natural host. When these hosts were challenged with pathogenic bacteria, mosquitoes in new host-Wolbachia symbioses had a higher survival rate than in old host-Wolbachia symbioses.

CONCLUSIONS/SIGNIFICANCE: The presence of Wolbachia per se does not necessarily affect pathogen infections, suggesting that a competition for limited nutrients is not the main reason for Wolbachia-mediated pathogen suppression. Instead, host immune responses are responsible for it. The elucidation of an immunity nature of PI is important to guide future practice: Wolbachia may be genetically engineered to be more immunogenic, it is desired to search and isolate more strains of Wolbachia, and test more host-Wolbachia symbioses for future applications. Our results also suggest Wolbachia-based PI may be applied to naturally Wolbachia-infected mosquito populations, and extend to the control of a broader range of mosquito-borne diseases.}, } @article {pmid32076535, year = {2020}, author = {Ross, PA and Callahan, AG and Yang, Q and Jasper, M and Arif, MAK and Afizah, AN and Nazni, WA and Hoffmann, AA}, title = {An elusive endosymbiont: Does Wolbachia occur naturally in Aedes aegypti?.}, journal = {Ecology and evolution}, volume = {10}, number = {3}, pages = {1581-1591}, pmid = {32076535}, issn = {2045-7758}, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria found within many insect species. Aedes mosquitoes experimentally infected with Wolbachia are being released into the field for Aedes-borne disease control. These Wolbachia infections induce cytoplasmic incompatibility which is used to suppress populations through incompatible matings or replace populations through the reproductive advantage provided by this mechanism. However, the presence of naturally occurring Wolbachia in target populations could interfere with both population replacement and suppression programs depending on the compatibility patterns between strains. Aedes aegypti were thought to not harbor Wolbachia naturally but several recent studies have detected Wolbachia in natural populations of this mosquito. We therefore review the evidence for natural Wolbachia infections in A. aegypti to date and discuss limitations of these studies. We draw on research from other mosquito species to outline the potential implications of natural Wolbachia infections in A. aegypti for disease control. To validate previous reports, we obtained a laboratory population of A. aegypti from New Mexico, USA, that harbors a natural Wolbachia infection, and we conducted field surveys in Kuala Lumpur, Malaysia, where a natural Wolbachia infection has also been reported. However, we were unable to detect Wolbachia in both the laboratory and field populations. Because the presence of naturally occurring Wolbachia in A. aegypti could have profound implications for Wolbachia-based disease control programs, it is important to continue to accurately assess the Wolbachia status of target Aedes populations.}, } @article {pmid32066253, year = {2020}, author = {Tan, FHP and Liu, G and Lau, SA and Jaafar, MH and Park, YH and Azzam, G and Li, Y and Liong, MT}, title = {Lactobacillus probiotics improved the gut microbiota profile of a Drosophila melanogaster Alzheimer's disease model and alleviated neurodegeneration in the eye.}, journal = {Beneficial microbes}, volume = {11}, number = {1}, pages = {79-89}, doi = {10.3920/BM2019.0086}, pmid = {32066253}, issn = {1876-2891}, mesh = {*Alzheimer Disease/drug therapy/microbiology ; Animals ; Biomarkers ; Disease Models, Animal ; Drosophila melanogaster ; Gastrointestinal Microbiome/*drug effects ; *Lactobacillus plantarum ; Neurodegenerative Diseases/drug therapy/*microbiology ; Probiotics/*administration & dosage/pharmacology ; Wolbachia/drug effects ; }, abstract = {Alzheimer's disease (AD) is a progressive disease and one of the most common forms of neurodegenerative disorders. Emerging evidence is supporting the use of various strategies that modulate gut microbiota to exert neurological and psychological changes. This includes the utilisation of probiotics as a natural and dietary intervention for brain health. Here, we showed the potential AD-reversal effects of Lactobacillus probiotics through feeding to our Drosophila melanogaster AD model. The administration of Lactobacillus strains was able to rescue the rough eye phenotype (REP) seen in AD-induced Drosophila, with a more prominent effect observed upon the administration of Lactobacillus plantarum DR7 (DR7). Furthermore, we analysed the gut microbiota of the AD-induced Drosophila and found elevated levels of Wolbachia. The administration of DR7 restored the gut microbiota diversity of AD-induced Drosophila with a significant reduction in Wolbachia's relative abundance, accompanied by an increase of Stenotrophomonas and Acetobacter. Through functional predictive analyses, Wolbachia was predicted to be positively correlated with neurodegenerative disorders, such as Parkinson's, Huntington's and Alzheimer's diseases, while Stenotrophomonas was negatively correlated with these neurodegenerative disorders. Altogether, our data exhibited DR7's ability to ameliorate the AD effects in our AD-induced Drosophila. Thus, we propose that Wolbachia be used as a potential biomarker for AD.}, } @article {pmid32058670, year = {2020}, author = {Madhav, M and Brown, G and Morgan, JA and Asgari, S and McGraw, EA and Munderloh, UG and Kurtti, TJ and James, P}, title = {Wolbachia successfully replicate in a newly established horn fly, Haematobia irritans irritans (L.) (Diptera: Muscidae) cell line.}, journal = {Pest management science}, volume = {76}, number = {7}, pages = {2441-2452}, doi = {10.1002/ps.5785}, pmid = {32058670}, issn = {1526-4998}, support = {//Meat and Livestock Australia/ ; }, mesh = {Animals ; Cell Line ; Drosophila melanogaster ; Insecticides ; *Muscidae ; *Wolbachia ; }, abstract = {BACKGROUND: Haematobia spp., horn flies (HF) and buffalo flies (BF), are economically important ectoparasites of dairy and beef cattle. Control of these flies relies mainly on treating cattle with chemical insecticides. However, the development of resistance to commonly used compounds is compromising the effectiveness of these treatments and alternative methods of control are required. Wolbachia are maternally transmitted endosymbiotic bacteria of arthropods that cause various reproductive distortions and fitness effects, making them a potential candidate for use in the biological control of pests. The first step towards this is the establishment and adaptation of xenobiotic infections of Wolbachia in target host cell lines.

RESULTS: Here, we report the successful establishment of a continuous HF cell line (HIE-18) from embryonic cells and its stable transinfection with Wolbachia strains wAlbB native to mosquitoes, and wMel and wMelPop native to Drosophila melanogaster. HIE-18 cells were typically round and diploid with ten chromosomes (2n = 10) or tetraploid with 20 chromosomes (4n = 20), with a doubling time of 67.2 h. Wolbachia density decreased significantly in HIE-18 cells in the first 48 h of infection, possibly due to overexpression of antimicrobial peptides through the Imd immune signalling pathway. However, density recovered after this time and HIE-18 cell lines stably infected with the three strains of Wolbachia have now each been subcultured more than 50 times as persistently infected lines.

CONCLUSION: The amenability of HF cells to infection with different strains of Wolbachia and the establishment of stable sustaining infections suggest the potential for use of Wolbachia in novel approaches for the control of Haematobia spp. Further, the availability of the HIE-18 cell line will provide an important resource for the study of genetics, host-parasite interactions and chemical resistance in Haematobia populations. © 2020 Society of Chemical Industry.}, } @article {pmid32053259, year = {2020}, author = {Vaught, RC and Voigt, S and Dobler, R and Clancy, DJ and Reinhardt, K and Dowling, DK}, title = {Interactions between cytoplasmic and nuclear genomes confer sex-specific effects on lifespan in Drosophila melanogaster.}, journal = {Journal of evolutionary biology}, volume = {33}, number = {5}, pages = {694-713}, doi = {10.1111/jeb.13605}, pmid = {32053259}, issn = {1420-9101}, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology ; Female ; Genetic Variation ; *Genome, Insect ; *Genome, Mitochondrial ; Longevity/*genetics ; Male ; Wolbachia ; }, abstract = {Genetic variation outside of the cell nucleus can affect the phenotype. The cytoplasm is home to the mitochondria, and in arthropods often hosts intracellular bacteria such as Wolbachia. Although numerous studies have implicated epistatic interactions between cytoplasmic and nuclear genetic variation as mediators of phenotypic expression, two questions remain. Firstly, it remains unclear whether outcomes of cyto-nuclear interactions will manifest differently across the sexes, as might be predicted given that cytoplasmic genomes are screened by natural selection only through females as a consequence of their maternal inheritance. Secondly, the relative contribution of mitochondrial genetic variation to other cytoplasmic sources of variation, such as Wolbachia infection, in shaping phenotypic outcomes of cyto-nuclear interactions remains unknown. Here, we address these questions, creating a fully crossed set of replicated cyto-nuclear populations derived from three geographically distinct populations of Drosophila melanogaster, measuring the lifespan of males and females from each population. We observed that cyto-nuclear interactions shape lifespan and that the outcomes of these interactions differ across the sexes. Yet, we found no evidence that placing the cytoplasms from one population alongside the nuclear background of others (generating putative cyto-nuclear mismatches) leads to decreased lifespan in either sex. Although it was difficult to partition mitochondrial from Wolbachia effects, our results suggest at least some of the cytoplasmic genotypic contribution to lifespan was directly mediated by an effect of sequence variation in the mtDNA. Future work should explore the degree to which cyto-nuclear interactions result in sex differences in the expression of other components of organismal life history.}, } @article {pmid32052099, year = {2020}, author = {Nakabachi, A and Malenovský, I and Gjonov, I and Hirose, Y}, title = {16S rRNA Sequencing Detected Profftella, Liberibacter, Wolbachia, and Diplorickettsia from Relatives of the Asian Citrus Psyllid.}, journal = {Microbial ecology}, volume = {80}, number = {2}, pages = {410-422}, doi = {10.1007/s00248-020-01491-z}, pmid = {32052099}, issn = {1432-184X}, support = {KAKENHI grant number 26292174//the Japan Society for the Promotion of Science/ ; NA//Tatematsu Foundation/ ; NA//Nagase Science and Technology Foundation/ ; }, mesh = {Animals ; Bacteria/classification/*isolation & purification ; Female ; France ; Hemiptera/*microbiology ; Male ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {The Asian citrus psyllid Diaphorina citri (Hemiptera: Psylloidea) is a serious pest of citrus species worldwide because it transmits Candidatus Liberibacter spp. (Alphaproteobacteria: Rhizobiales), the causative agents of the incurable citrus disease, huanglongbing or greening disease. Diaphorina citri possesses a specialized organ called a bacteriome, which harbors vertically transmitted intracellular mutualists, Ca. Carsonella ruddii (Gammaproteobacteria: Oceanospirillales) and Ca. Profftella armatura (Gammaproteobacteria: Betaproteobacteriales). Whereas Carsonella is a typical nutritional symbiont, Profftella is an unprecedented type of toxin-producing defensive symbiont, unusually sharing organelle-like features with nutritional symbionts. Additionally, many D. citri strains are infected with Wolbachia, which manipulate reproduction in various arthropod hosts. In the present study, in an effort to obtain insights into the evolution of symbioses between Diaphorina and bacteria, microbiomes of psyllids closely related to D. citri were investigated. Bacterial populations of Diaphorina cf. continua and Diaphorina lycii were analyzed using Illumina sequencing of 16S rRNA gene amplicons and compared with data obtained from D. citri. The analysis revealed that all three Diaphorina spp. harbor Profftella as well as Carsonella lineages, implying that Profftella is widespread within the genus Diaphorina. Moreover, the analysis identified Ca. Liberibacter europaeus and Diplorickettsia sp. (Gammaproteobacteria: Diplorickettsiales) in D. cf. continua, and a total of four Wolbachia (Alphaproteobacteria: Rickettsiales) lineages in the three psyllid species. These results provide deeper insights into the interactions among insects, bacteria, and plants, which would eventually help to better manage horticulture.}, } @article {pmid32047292, year = {2020}, author = {Zélé, F and Santos, I and Matos, M and Weill, M and Vavre, F and Magalhães, S}, title = {Endosymbiont diversity in natural populations of Tetranychus mites is rapidly lost under laboratory conditions.}, journal = {Heredity}, volume = {124}, number = {4}, pages = {603-617}, pmid = {32047292}, issn = {1365-2540}, mesh = {Animals ; *Bacteroidetes/genetics ; Female ; Laboratories ; *Rickettsia/genetics ; *Symbiosis ; *Tetranychidae/microbiology ; *Wolbachia/genetics ; }, abstract = {Although the diversity of bacterial endosymbionts in arthropods is well documented, whether and how such diversity is maintained remains an open question. We investigated the temporal changes occurring in the prevalence and composition of endosymbionts after transferring natural populations of Tetranychus spider mites from the field to the laboratory. These populations, belonging to three different Tetranychus species (T. urticae, T. ludeni and T. evansi) carried variable infection frequencies of Wolbachia, Cardinium, and Rickettsia. We report a rapid change of the infection status of these populations after only 6 months of laboratory rearing, with an apparent loss of Rickettsia and Cardinium, while Wolbachia apparently either reached fixation or was lost. We show that Wolbachia had variable effects on host longevity and fecundity, and induced variable levels of cytoplasmic incompatibility (CI) in each fully infected population, despite no sequence divergence in the markers used and full CI rescue between all populations. This suggests that such effects are largely dependent upon the host genotype. Subsequently, we used these data to parameterize a theoretical model for the invasion of CI-inducing symbionts in haplodiploids, which shows that symbiont effects are sufficient to explain their dynamics in the laboratory. This further suggests that symbiont diversity and prevalence in the field are likely maintained by environmental heterogeneity, which is reduced in the laboratory. Overall, this study highlights the lability of endosymbiont infections and draws attention to the limitations of laboratory studies to understand host-symbiont interactions in natural populations.}, } @article {pmid32043447, year = {2020}, author = {Lechner, AM and Gastager, H and Kern, JM and Wagner, B and Tappe, D}, title = {Case Report: Successful Treatment of a Patient with Microfilaremic Dirofilariasis Using Doxycycline.}, journal = {The American journal of tropical medicine and hygiene}, volume = {102}, number = {4}, pages = {844-846}, pmid = {32043447}, issn = {1476-1645}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; *Dirofilaria repens ; Dirofilariasis/*diagnostic imaging/*therapy ; Doxycycline/*therapeutic use ; Female ; Humans ; Middle Aged ; }, abstract = {We report the case of a 56-year-old woman with microfilaremic dirofilariasis due to Dirofilaria repens, which is a very rare condition in humans. Of note, just one of six large-volume blood samples of this patient was positive for microfilariae. Polymerase chain reaction (PCR) and sequencing of the parasite gene determined the geographic origin of the causative helminth. The patient was treated successfully with doxycycline. This drug was chosen because of the patient's reluctance to the use of ivermectin and to provide an anthelmintic effect by targeting the bacterial endosymbiont Wolbachia present in most filarial species.}, } @article {pmid32041638, year = {2020}, author = {Ekwudu, O and Devine, GJ and Aaskov, JG and Frentiu, FD}, title = {Wolbachia strain wAlbB blocks replication of flaviviruses and alphaviruses in mosquito cell culture.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {54}, pmid = {32041638}, issn = {1756-3305}, support = {APP1020817//National Health and Medical Research Council (NHMRC) of Australia/ ; Internal grant//Queensland University of Technology/ ; }, mesh = {Aedes/microbiology/virology ; Alphavirus/*growth & development ; Alphavirus Infections/prevention & control ; Animals ; Cell Line/microbiology/virology ; Dengue/prevention & control ; Flavivirus/*growth & development ; Humans ; Insect Vectors/microbiology/virology ; *Microbial Interactions ; Pest Control, Biological ; Virus Diseases/prevention & control/transmission ; *Virus Replication ; West Nile Fever/prevention & control ; *Wolbachia ; Zika Virus Infection/prevention & control ; }, abstract = {BACKGROUND: Wolbachia pipientis are bacterial endosymbionts of arthropods currently being implemented as biocontrol agents to reduce the global burden of arboviral diseases. Some strains of Wolbachia, when introduced into Aedes aegypti mosquitoes, reduce or block the replication of RNA viruses pathogenic to humans. The wAlbB strain of Wolbachia was originally isolated from Aedes albopictus, and when transinfected into Ae. aegypti, persists in mosquitoes under high temperature conditions longer than other strains. The utility of wAlbB to block a broad spectrum of RNA viruses has received limited attention. Here we test the ability of wAlbB to reduce or block the replication of a range of Flavivirus and Alphavirus species in cell culture.

METHODS: The C6/36 mosquito cell line was stably infected with the wAlbB strain using the shell-vial technique. The replication of dengue, West Nile and three strains of Zika (genus Flavivirus), and Ross River, Barmah Forest and Sindbis (genus Alphavirus) viruses was compared in wAlbB-infected cells with Wolbachia-free controls. Infectious virus titres were determined using either immunofocus or plaque assays. A general linear model was used to test for significant differences in replication between flaviviruses and alphaviruses.

RESULTS: Titres of all viruses were significantly reduced in cell cultures infected with wAlbB versus Wolbachia-free controls. The magnitude of reduction in virus yields varied among virus species and, within species, also among the strains utilized.

CONCLUSION: Our results suggest that wAlbB infection of arthropods could be used to reduce transmission of a wide range of pathogenic RNA viruses.}, } @article {pmid32041484, year = {2020}, author = {Nazareth, T and Craveiro, I and Moutinho, A and Seixas, G and Gonçalves, C and Gonçalves, L and Teodósio, R and Sousa, CA}, title = {What happens when we modify mosquitoes for disease prevention? A systematic review.}, journal = {Emerging microbes & infections}, volume = {9}, number = {1}, pages = {348-365}, pmid = {32041484}, issn = {2222-1751}, mesh = {Animals ; *Animals, Genetically Modified ; Gene Transfer, Horizontal ; Mosquito Control/*methods ; Mosquito Vectors/*genetics/microbiology ; Wolbachia ; }, abstract = {The release of modified mosquitoes to suppress/replace vectors constitutes a promising tool for vector control and disease prevention. Evidence regarding these innovative modification techniques is scarce and disperse. This work conducted a systematic review, gathering and analysing research articles from PubMed and Biblioteca Virtual em Saúde databases whose results report efficacy and non-target effects of using modified insects for disease prevention, until 2016. More than 1500 publications were screened and 349 were analysed. Only 12/3.4% articles reported field-based evidence and 41/11.7% covered modification strategies' post-release efficacy. Variability in the effective results (90/25.7%) questioned its reproducibility in different settings. We also found publications reporting reversal outcomes 38/10.9%, (e.g. post-release increase of vector population). Ecological effects were also reported, such as horizontal transfer events (54/15.5%), and worsening pathogenesis induced by natural wolbachia (10/2.9%). Present work revealed promising outcomes of modifying strategies. However, it also revealed a need for field-based evidence mainly regarding epidemiologic and long-term impact. It pointed out some eventual irreversible and important effects that must not be ignored when considering open-field releases, and that may constitute constraints to generate the missing field evidence. Present work constitutes a baseline of knowledge, offering also a methodological approach that may facilitate future updates.}, } @article {pmid32040182, year = {2020}, author = {Shang, F and Niu, J and Ding, BY and Wang, JJ}, title = {Comparative Insight into the Bacterial Communities in Alate and Apterous Morphs of Brown Citrus Aphid (Hemiptera: Aphididae).}, journal = {Journal of economic entomology}, volume = {113}, number = {3}, pages = {1436-1444}, doi = {10.1093/jee/toaa016}, pmid = {32040182}, issn = {1938-291X}, mesh = {Animals ; *Aphids/genetics ; Bacteria ; *Buchnera ; *Citrus ; RNA, Ribosomal, 16S/genetics ; }, abstract = {Wing polyphenism (alate and apterous morphs) in aphids is a trade-off between dispersal and reproduction. How bacterial communities are associated with wing polyphenism in aphids is still not clearly understood. This study used 16S rRNA sequencing to examine the differences in diversity of the bacterial community between alate and apterous morphs in Aphis citricidus, the main vector of the Citrus tristeza virus. Eighty-one operational taxonomic units (OTUs) belonging to 37 orders, 34 classes, and 13 phyla were identified from all samples. Among these OTUs, Wolbachia (79.17%), Buchnera (17.64%), and Pseudomonas (2.99%) were the dominant bacterial genera. The diversity of symbionts varied between the two morphs; apterous morphs had more bacterial diversity (69 OTUs belonging to 45 families, 21 classes, and 12 phyla) than alate morphs (45 OTUs belonging to 36 families, 15 classes, and 10 phyla). In addition, the abundance of five OTUs was significantly different between two morphs. Among these OTUs, two Pseudomonas species (Pseudomonas_brenneri [OTU21] and unclassified_Pseudomonas [OTU13]) represented a high proportion (3.93% and 2.06%) in alate morphs but were present in low abundance (0.006% and 0.002%) in apterous morphs. RT-qPCR showed consistent results with high-throughput DNA sequencing. The preliminary survey showed the difference in composition and frequency of bacteria between alate and apterous morphs. Thus, the results contribute to anew insight of microorganisms that may be involved in wing dimorphism and helpful for controlling the dispersal of this pest through artificial elimination or reinfection of bacterial symbionts or targeting symbiosis-related host genes by RNA interference in future.}, } @article {pmid32030839, year = {2020}, author = {Hundertmark, A and Goodacre, SL and Brookfield, JFY}, title = {Alternative evolutionary outcomes following endosymbiont-mediated selection on male mating preference alleles.}, journal = {Journal of evolutionary biology}, volume = {33}, number = {5}, pages = {653-667}, pmid = {32030839}, issn = {1420-9101}, mesh = {Animals ; Arthropods/*microbiology ; Avoidance Learning ; *Biological Evolution ; Female ; Male ; *Mating Preference, Animal ; *Models, Biological ; Selection, Genetic ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {In many arthropods, intracellular bacteria, such as those of the genus Wolbachia, may spread through host populations as a result of cytoplasmic incompatibility (CI). Here, there is sterility or reduced fertility in crosses between infected males and uninfected females. As the bacterium is maternally inherited, the reduced fertility of uninfected females increases the frequency of the infection. If the transmission fidelity of the bacterium is less than 100%, the bacterium cannot invade from a low frequency, but if its frequency exceeds a threshold, it increases to a high, stable, equilibrium frequency. We explore the expected evolutionary dynamics of mutant alleles that cause their male bearers to avoid mating with uninfected females. For alleles which create this avoidance behaviour conditional upon the male being infected, there is a wide zone of parameter space that allows the preference allele to drive Wolbachia from the population when it would otherwise stably persist. There is also a wide zone of parameter space that allows a joint stable equilibrium for the Wolbachia and a polymorphism for the preference allele. When the male's avoidance of uninfected females is unconditional, the preference allele's effect on Wolbachia frequency is reduced, but there is a narrow range of values for the transmission rate and CI fertility that allow an unconditional preference allele to drive Wolbachia from the population, in a process driven by positive linkage disequilibrium between Wolbachia and the preference allele. The possibility of the evolution of preference could hamper attempts to manipulate wild populations through Wolbachia introductions.}, } @article {pmid32028994, year = {2020}, author = {Uni, S and Mat Udin, AS and Agatsuma, T and Junker, K and Saijuntha, W and Bunchom, N and Fukuda, M and Martin, C and Lefoulon, E and Labat, A and Khan, FAA and Low, VL and Cheah, PL and Lim, YA and Ramli, R and Belabut, DM and Zainuri, NA and Matsubayashi, M and Omar, H and Bhassu, S and Uga, S and Hashim, R and Takaoka, H and Azirun, MS}, title = {Description, molecular characteristics and Wolbachia endosymbionts of Onchocerca borneensis Uni, Mat Udin & Takaoka n. sp. (Nematoda: Filarioidea) from the Bornean bearded pig Sus barbatus Müller (Cetartiodactyla: Suidae) of Sarawak, Malaysia.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {50}, pmid = {32028994}, issn = {1756-3305}, support = {FRGS EP020-2012//The Ministry of Higher Education, Malaysia/ ; }, mesh = {Animals ; Biological Coevolution ; Classification ; Genes, Bacterial ; Genes, Helminth ; Humans ; *Onchocerca/anatomy & histology/classification/microbiology ; Onchocerciasis/transmission/*veterinary ; Onchocerciasis, Ocular/parasitology/transmission ; Phylogeny ; Swine/*parasitology ; Swine Diseases ; Symbiosis ; *Wolbachia/classification/isolation & purification ; Zoonoses/transmission ; }, abstract = {BACKGROUND: The genus Onchocerca Diesing, 1841 includes species of medical importance, such as O. volvulus (Leuckart, 1893), which causes river blindness in the tropics. Recently, zoonotic onchocercosis has been reported in humans worldwide. In Japan, O. dewittei japonica Uni, Bain & Takaoka, 2001 from wild boars is a causative agent for this zoonosis. Many filarioid nematodes are infected with Wolbachia endosymbionts which exhibit various evolutionary relationships with their hosts. While investigating the filarial fauna of Borneo, we discovered an undescribed Onchocerca species in the bearded pig Sus barbatus Müller (Cetartiodactyla: Suidae).

METHODS: We isolated Onchocerca specimens from bearded pigs and examined their morphology. For comparative material, we collected fresh specimens of O. d. dewittei Bain, Ramachandran, Petter & Mak, 1977 from banded pigs (S. scrofa vittatus Boie) in Peninsular Malaysia. Partial sequences of three different genes (two mitochondrial genes, cox1 and 12S rRNA, and one nuclear ITS region) of these filarioids were analysed. By multi-locus sequence analyses based on six genes (16S rDNA, ftsZ, dnaA, coxA, fbpA and gatB) of Wolbachia, we determined the supergroups in the specimens from bearded pigs and those of O. d. dewittei.

RESULTS: Onchocerca borneensis Uni, Mat Udin & Takaoka n. sp. is described on the basis of morphological characteristics and its genetic divergence from congeners. Molecular characteristics of the new species revealed its close evolutionary relationship with O. d. dewittei. Calculated p-distance for the cox1 gene sequences between O. borneensis n. sp. and O. d. dewittei was 5.9%, while that between O. d. dewittei and O. d. japonica was 7.6%. No intraspecific genetic variation was found for the new species. Wolbachia strains identified in the new species and O. d. dewittei belonged to supergroup C and are closely related.

CONCLUSIONS: Our molecular analyses of filarioids from Asian suids indicate that the new species is sister to O. d. dewittei. On the basis of its morphological and molecular characteristics, we propose to elevate O. d. japonica to species level as O. japonica Uni, Bain & Takaoka, 2001. Coevolutionary relationships exist between the Wolbachia strains and their filarial hosts in Borneo and Peninsular Malaysia.}, } @article {pmid32024068, year = {2020}, author = {de Jesus, CP and Dias, FBS and Villela, DMA and Maciel-de-Freitas, R}, title = {Ovitraps Provide a Reliable Estimate of Wolbachia Frequency during wMelBr Strain Deployment in a Geographically Isolated Aedes aegypti Population.}, journal = {Insects}, volume = {11}, number = {2}, pages = {}, pmid = {32024068}, issn = {2075-4450}, support = {001//Coordenação de Aperfeiçoamento de Pessoal de Nível Superior/ ; }, abstract = {Deployment of Aedes aegypti mosquitoes carrying the endosymbiont bacterium Wolbachia has been identified as a promising strategy to reduce dengue, chikungunya, and Zika transmission. We investigated whether sampling larvae from ovitraps can provide reliable estimates on Wolbachia frequency during releases, as compared to the expensive adult-based BG-Sentinel. We conducted pilot releases in a semi-field system (SFS) divided into six cages of 21 m[2], each with five ovitraps. Five treatments were chosen to represent different points of a hypothetical invasion curve: 10%, 25%, 50%, 75%, and 90% of Wolbachia frequency. Collected eggs were counted and hatched, and the individuals from a net sample of 27% of larvae per treatment were screened for Wolbachia presence by RT-qPCR. Ovitrap positioning had no effect on egg hatching rate. Treatment strongly affected the number of eggs collected and also the hatching rate, especially when Wolbachia was at a 10% frequency. A second observation was done during the release of Wolbachia in Rio under a population replacement approach when bacterium frequency was estimated using 30 BG-Sentinel traps and 45 ovitraps simultaneously. By individually screening 35% (N = 3904) of larvae collected by RT-qPCR, we were able to produce a similar invasion curve to the one observed when all adults were individually screened. If sampling is reduced to 20%, monitoring Wolbachia frequency with 45 ovitraps would be roughly half the cost of screening all adult mosquitoes captured by 30 BG-Sentinels. Our findings support the scale-up of Wolbachia releases, especially in areas with limited resources to afford massive trapping with BG-Sentinel traps.}, } @article {pmid32014020, year = {2020}, author = {Woodhams, DC and Bletz, MC and Becker, CG and Bender, HA and Buitrago-Rosas, D and Diebboll, H and Huynh, R and Kearns, PJ and Kueneman, J and Kurosawa, E and LaBumbard, BC and Lyons, C and McNally, K and Schliep, K and Shankar, N and Tokash-Peters, AG and Vences, M and Whetstone, R}, title = {Host-associated microbiomes are predicted by immune system complexity and climate.}, journal = {Genome biology}, volume = {21}, number = {1}, pages = {23}, pmid = {32014020}, issn = {1474-760X}, mesh = {Adaptation, Physiological ; Animals ; *Climate ; Host-Pathogen Interactions/*immunology ; Humans ; *Microbiota ; }, abstract = {BACKGROUND: Host-associated microbiomes, the microorganisms occurring inside and on host surfaces, influence evolutionary, immunological, and ecological processes. Interactions between host and microbiome affect metabolism and contribute to host adaptation to changing environments. Meta-analyses of host-associated bacterial communities have the potential to elucidate global-scale patterns of microbial community structure and function. It is possible that host surface-associated (external) microbiomes respond more strongly to variations in environmental factors, whereas internal microbiomes are more tightly linked to host factors.

RESULTS: Here, we use the dataset from the Earth Microbiome Project and accumulate data from 50 additional studies totaling 654 host species and over 15,000 samples to examine global-scale patterns of bacterial diversity and function. We analyze microbiomes from non-captive hosts sampled from natural habitats and find patterns with bioclimate and geophysical factors, as well as land use, host phylogeny, and trophic level/diet. Specifically, external microbiomes are best explained by variations in mean daily temperature range and precipitation seasonality. In contrast, internal microbiomes are best explained by host factors such as phylogeny/immune complexity and trophic level/diet, plus climate.

CONCLUSIONS: Internal microbiomes are predominantly associated with top-down effects, while climatic factors are stronger determinants of microbiomes on host external surfaces. Host immunity may act on microbiome diversity through top-down regulation analogous to predators in non-microbial ecosystems. Noting gaps in geographic and host sampling, this combined dataset represents a global baseline available for interrogation by future microbial ecology studies.}, } @article {pmid31996463, year = {2020}, author = {Cattarino, L and Rodriguez-Barraquer, I and Imai, N and Cummings, DAT and Ferguson, NM}, title = {Mapping global variation in dengue transmission intensity.}, journal = {Science translational medicine}, volume = {12}, number = {528}, pages = {}, doi = {10.1126/scitranslmed.aax4144}, pmid = {31996463}, issn = {1946-6242}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; U01 CK000510/CK/NCEZID CDC HHS/United States ; U01 GM110721/GM/NIGMS NIH HHS/United States ; }, mesh = {Basic Reproduction Number ; Cost of Illness ; Dengue/*epidemiology/immunology/*transmission/virology ; Dengue Vaccines/immunology ; Geography ; Humans ; *Internationality ; Risk Factors ; }, abstract = {Intervention planning for dengue requires reliable estimates of dengue transmission intensity. However, current maps of dengue risk provide estimates of disease burden or the boundaries of endemicity rather than transmission intensity. We therefore developed a global high-resolution map of dengue transmission intensity by fitting environmentally driven geospatial models to geolocated force of infection estimates derived from cross-sectional serological surveys and routine case surveillance data. We assessed the impact of interventions on dengue transmission and disease using Wolbachia-infected mosquitoes and the Sanofi-Pasteur vaccine as specific examples. We predicted high transmission intensity in all continents straddling the tropics, with hot spots in South America (Colombia, Venezuela, and Brazil), Africa (western and central African countries), and Southeast Asia (Thailand, Indonesia, and the Philippines). We estimated that 105 [95% confidence interval (CI), 95 to 114] million dengue infections occur each year with 51 (95% CI, 32 to 66) million febrile disease cases. Our analysis suggests that transmission-blocking interventions such as Wolbachia, even at intermediate efficacy (50% transmission reduction), might reduce global annual disease incidence by up to 90%. The Sanofi-Pasteur vaccine, targeting only seropositive recipients, might reduce global annual disease incidence by 20 to 30%, with the greatest impact in high-transmission settings. The transmission intensity map presented here, and made available for download, may help further assessment of the impact of dengue control interventions and prioritization of global public health efforts.}, } @article {pmid31986143, year = {2020}, author = {Ehrens, A and Lunde, CS and Jacobs, RT and Struever, D and Koschel, M and Frohberger, SJ and Lenz, F and Fendler, M and Turner, JD and Ward, SA and Taylor, MJ and Freund, YR and Stefanakis, R and Easom, E and Li, X and Plattner, JJ and Hoerauf, A and Hübner, MP}, title = {In vivo efficacy of the boron-pleuromutilin AN11251 against Wolbachia of the rodent filarial nematode Litomosoides sigmodontis.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {1}, pages = {e0007957}, pmid = {31986143}, issn = {1935-2735}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Boron ; Diterpenes/*pharmacology ; Doxycycline/pharmacology ; Female ; Filariasis/*drug therapy/microbiology ; Filarioidea/*drug effects/microbiology ; Mice, Inbred BALB C ; Polycyclic Compounds/*pharmacology ; Rifampin/pharmacology ; Symbiosis ; Wolbachia/*drug effects ; Pleuromutilins ; }, abstract = {The elimination of filarial diseases such as onchocerciasis and lymphatic filariasis is hampered by the lack of a macrofilaricidal-adult worm killing-drug. In the present study, we tested the in vivo efficacy of AN11251, a boron-pleuromutilin that targets endosymbiotic Wolbachia bacteria from filarial nematodes and compared its efficacy to doxycycline and rifampicin. Doxycycline and rifampicin were previously shown to deplete Wolbachia endosymbionts leading to a permanent sterilization of the female adult filariae and adult worm death in human clinical studies. Twice-daily oral treatment of Litomosoides sigmodontis-infected mice with 200 mg/kg AN11251 for 10 days achieved a Wolbachia depletion > 99.9% in the adult worms, exceeding the Wolbachia reduction by 10-day treatments with bioequivalent human doses of doxycycline and a similar reduction as high-dose rifampicin (35 mg/kg). Wolbachia reductions of > 99% were also accomplished by 14 days of oral AN11251 at a lower twice-daily dose (50 mg/kg) or once-per-day 200 mg/kg AN11251 treatments. The combinations tested of AN11251 with doxycycline had no clear beneficial impact on Wolbachia depletion, achieving a > 97% Wolbachia reduction with 7 days of treatment. These results indicate that AN11251 is superior to doxycycline and comparable to high-dose rifampicin in the L. sigmodontis mouse model, allowing treatment regimens as short as 10-14 days. Therefore, AN11251 represents a promising pre-clinical candidate that was identified in the L. sigmodontis model, and could be further evaluated and developed as potential clinical candidate for human lymphatic filariasis and onchocerciasis.}, } @article {pmid31971938, year = {2020}, author = {Ross, PA and Axford, JK and Yang, Q and Staunton, KM and Ritchie, SA and Richardson, KM and Hoffmann, AA}, title = {Heatwaves cause fluctuations in wMel Wolbachia densities and frequencies in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {1}, pages = {e0007958}, pmid = {31971938}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Australia ; Female ; Host-Pathogen Interactions ; *Hot Temperature ; Wolbachia/classification/*physiology ; }, abstract = {Aedes aegypti mosquitoes infected with the wMel strain of Wolbachia are being released into natural mosquito populations in the tropics as a way of reducing dengue transmission. High temperatures adversely affect wMel, reducing Wolbachia density and cytoplasmic incompatibility in some larval habitats that experience large temperature fluctuations. We monitored the impact of a 43.6°C heatwave on the wMel infection in a natural population in Cairns, Australia, where wMel was first released in 2011 and has persisted at a high frequency. Wolbachia infection frequencies in the month following the heatwave were reduced to 83% in larvae sampled directly from field habitats and 88% in eggs collected from ovitraps, but recovered to be near 100% four months later. Effects of the heatwave on wMel appeared to be stage-specific and delayed, with reduced frequencies and densities in field-collected larvae and adults reared from ovitraps but higher frequencies in field-collected adults. Laboratory experiments showed that the effects of heatwaves on cytoplasmic incompatibility and density are life stage-specific, with first instar larvae being the most vulnerable to temperature effects. Our results indicate that heatwaves in wMel-infected populations will have only temporary effects on Wolbachia frequencies and density once the infection has established in the population. Our results are relevant to ongoing releases of wMel-infected Ae. aegypti in several tropical countries.}, } @article {pmid31961929, year = {2020}, author = {Covey, H and Hall, RH and Krafsur, A and Matthews, ML and Shults, PT and Brelsfoard, CL}, title = {Cryptic Wolbachia (Rickettsiales: Rickettsiaceae) Detection and Prevalence in Culicoides (Diptera: Ceratopogonidae) Midge Populations in the United States.}, journal = {Journal of medical entomology}, volume = {57}, number = {4}, pages = {1262-1269}, doi = {10.1093/jme/tjaa003}, pmid = {31961929}, issn = {1938-2928}, mesh = {Animals ; Ceratopogonidae/*microbiology ; Female ; In Situ Hybridization, Fluorescence ; Male ; Real-Time Polymerase Chain Reaction ; United States ; Wolbachia/classification/*isolation & purification ; }, abstract = {Culicoides midges vector numerous veterinary and human pathogens. Many of these diseases lack effective therapeutic treatments or vaccines to limit transmission. The only effective approach to limit disease transmission is vector control. However, current vector control for Culicoides midges is complicated by the biology of many Culicoides species and is not always effective at reducing midge populations and impacting disease transmission. The endosymbiont Wolbachia pipientis Hertig may offer an alternative control approach to limit disease transmission and affect Culicoides populations. Here the detection of Wolbachia infections in nine species of Culicoides midges is reported. Infections were detected at low densities using qPCR. Wolbachia infections were confirmed with the sequencing of a partial region of the 16S gene. Fluorescence in situ hybridization of Culicoides sonorensis Wirth and Jones adults and dissected ovaries confirm the presence of Wolbachia infections in an important vector of Bluetongue and Epizootic hemorrhagic disease viruses. The presence of Wolbachia in Culicoides populations in the United States suggests the need for further investigation of Wolbachia as a strategy to limit transmission of diseases vectored by Culicoides midges.}, } @article {pmid31958110, year = {2020}, author = {Bing, XL and Zhao, DS and Sun, JT and Zhang, KJ and Hong, XY}, title = {Genomic Analysis of Wolbachia from Laodelphax striatellus (Delphacidae, Hemiptera) Reveals Insights into Its "Jekyll and Hyde" Mode of Infection Pattern.}, journal = {Genome biology and evolution}, volume = {12}, number = {2}, pages = {3818-3831}, pmid = {31958110}, issn = {1759-6653}, mesh = {Animals ; Bacterial Proteins/genetics ; Biotin/metabolism ; Genome, Bacterial/genetics ; Genomics/methods ; Hemiptera/*microbiology ; Phenotype ; Phylogeny ; Riboflavin/metabolism ; Wolbachia/*genetics ; }, abstract = {Wolbachia is a widely distributed intracellular bacterial endosymbiont among invertebrates. The wStriCN, the Wolbachia strain that naturally infects an agricultural pest Laodelphax striatellus, has a "Jekyll and Hyde" mode of infection pattern with positive and negative effects: It not only kills many offspring by inducing cytoplasmic incompatibility (CI) but also significantly increases host fecundity. In this study, we assembled the draft genome of wStriCN and compared it with other Wolbachia genomes to look for clues to its Jekyll and Hyde characteristics. The assembled wStriCN draft genome is 1.79 Mb in size, which is the largest Wolbachia genome in supergroup B. Phylogenomic analysis showed that wStriCN is closest to Wolbachia from Asian citrus psyllid Diaphorina citri. These strains formed a monophylogentic clade within supergroup B. Compared with other Wolbachia genomes, wStriCN contains the most diverse insertion sequence families, the largest amount of prophage sequences, and the most ankyrin domain protein coding genes. The wStriCN genome encodes components of multiple secretion systems, including Types I, II, IV, VI, Sec, and Tac. We detected three pairs of homologs for CI factors CifA and CifB. These proteins harbor the catalytic domains responsible for CI phenotypes but are phylogenetically and structurally distinct from all known Cif proteins. The genome retains pathways for synthesizing biotin and riboflavin, which may explain the beneficial roles of wStriCN in its host planthoppers, which feed on nutrient-poor plant sap. Altogether, the genomic sequencing of wStriCN provides insight into understanding the phylogeny and biology of Wolbachia.}, } @article {pmid31956742, year = {2020}, author = {Zhang, H and Lui, R}, title = {Releasing Wolbachia-infected Aedes aegypti to prevent the spread of dengue virus: A mathematical study.}, journal = {Infectious Disease Modelling}, volume = {5}, number = {}, pages = {142-160}, pmid = {31956742}, issn = {2468-0427}, abstract = {Wolbachia is a bacterium that is present in 60% of insects but it is not generally found in Aedes aegypti, the primary vector responsible for the transmission of dengue virus, Zika virus, and other human diseases caused by RNA viruses. Wolbachia has been shown to stop the growth of a variety of RNA viruses in Drosophila and in mosquitoes. Wolbachia-infected Ae. aegypti have both reproductive advantages and disadvantages over wild types. If Wolbachia-infected females are fertilized by either normal or infected males, the offspring are healthy and Wolbachia-positive. On the other hand, if Wolbachia-negative females are fertilized by Wolbachia-positive males, the offspring do not hatch. This phenomenon is called cytoplasmic incompatibility. Thus, Wolbachia-positive females have a reproductive advantage, and the Wolbachia is expanded in the population. On the other hand, Wolbachia-infected mosquitoes lay fewer eggs and generally have a shorter lifespan. In recent years, scientists have successfully released these Wolbachia-adapted mosquitoes into the wild in several countries and have achieved a high level of replacement with Wolbachia-positive mosquitoes. Here, we propose a minimal mathematical model to investigate the feasibility of such a release method. The model has five steady-states two of which are locally asymptotically stable. One of these stable steady-states has no Wolbachia-infected mosquitoes while for the other steady-state, all mosquitoes are infected with Wolbachia. We apply optimal control theory to find a release method that will drive the mosquito population close to the steady-state with only Wolbachia-infected mosquitoes in a two-year time period. Because some of the model parameters cannot be accurately measured or predicted, we also perform uncertainty and sensitivity analysis to quantify how variations in our model parameters affect our results.}, } @article {pmid31942975, year = {2020}, author = {Zhu, YX and Song, ZR and Huo, SM and Yang, K and Hong, XY}, title = {Variation in the microbiome of the spider mite Tetranychus truncatus with sex, instar and endosymbiont infection.}, journal = {FEMS microbiology ecology}, volume = {96}, number = {2}, pages = {}, doi = {10.1093/femsec/fiaa004}, pmid = {31942975}, issn = {1574-6941}, mesh = {Animals ; Bacteria/genetics ; Female ; Fertility ; Male ; *Microbiota ; RNA, Ribosomal, 16S ; Spiroplasma/genetics ; *Symbiosis ; Tetranychidae/*microbiology ; Wolbachia/genetics ; }, abstract = {Most arthropod-associated bacterial communities play a crucial role in host functional traits, whose structure could be dominated by endosymbionts. The spider mite Tetranychus truncatus is a notorious agricultural pest harboring various endosymbionts, yet the effects of endosymbionts on spider mite microbiota remain largely unknown. Here, using deep sequencing of the 16S rRNA gene, we characterized the microbiota of male and female T. truncatus with different endosymbionts (Wolbachia and Spiroplasma) across different developmental stages. Although the spider mite microbiota composition varied across the different developmental stages, Proteobacteria were the most dominant bacteria harbored in all samples. Positive relationships among related operational taxonomic units dominated the significant coassociation networks among bacteria. Moreover, the spider mites coinfected with Wolbachia and Spiroplasma had a significantly higher daily fecundity and juvenile survival rate than the singly infected or uninfected spider mites. The possible function of spider-mite associated bacteria was discussed. Our results highlight the dynamics of spider mite microbiotas across different life stages, and the potential role of endosymbionts in shaping the microbiota of spider mites and improving host fitness.}, } @article {pmid31937677, year = {2020}, author = {Perlmutter, JI and Meyers, JE and Bordenstein, SR}, title = {Transgenic Testing Does Not Support a Role for Additional Candidate Genes in Wolbachia Male Killing or Cytoplasmic Incompatibility.}, journal = {mSystems}, volume = {5}, number = {1}, pages = {}, pmid = {31937677}, issn = {2379-5077}, support = {F31 AI143152/AI/NIAID NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; R21 AI133522/AI/NIAID NIH HHS/United States ; }, abstract = {Endosymbiotic bacteria in the genus Wolbachia remarkably infect nearly half of all arthropod species. They spread in part because of manipulations of host sexual reproduction that enhance the maternal transmission of the bacteria, including male killing (death of infected males) and unidirectional cytoplasmic incompatibility (CI; death of offspring from infected fathers and uninfected mothers). Recent discoveries identified several genes in prophage WO of Wolbachia (wmk, cifA, and cifB) that fully or partially recapitulate male killing or CI when transgenically expressed in Drosophila melanogaster However, it is not yet fully resolved if other gene candidates contribute to these phenotypes. Here, we transgenically tested 10 additional gene candidates for their involvement in male killing and/or CI. The results show that despite sequence and protein architecture similarities or comparative associations with reproductive parasitism, transgenic expression of the candidates does not recapitulate male killing or CI. Sequence analysis across Wmk and its closest relatives reveals amino acids that may be important to its function. In addition, evidence is presented to propose new hypotheses regarding the relationship between wmk transcript length and its ability to kill a given host, as well as copy number of wmk homologs within a bacterial strain, which may be predictive of host resistance. Together, these analyses continue to build the evidence for identification of wmk, cifA, and cifB as the major genes that have thus far been shown to cause reproductive parasitism in Wolbachia, and the transgenic resources provide a basis for further functional study of phage WO genes.IMPORTANCE Wolbachia are widespread bacterial endosymbionts that manipulate the reproduction of diverse arthropods to spread through a population and can substantially shape host evolution. Recently, reports identified three prophage WO genes (wmk, cifA, and cifB) that transgenically recapitulate many aspects of reproductive manipulation in Drosophila melanogaster Here, we transgenically tested 10 additional gene candidates for CI and/or male killing in flies. The results yield no evidence for the involvement of these gene candidates in reproductive parasitism, bolstering the evidence for identification of the cif and wmk genes as the major factors involved in their phenotypes. In addition, evidence supports new hypotheses for prediction of male-killing phenotypes or lack thereof based on wmk transcript length and copy number. These experiments inform efforts to understand the full basis of reproductive parasitism for basic and applied purposes and lay the foundation for future work on the function of an interesting group of Wolbachia and phage WO genes.}, } @article {pmid31937373, year = {2020}, author = {Hu, Y and Xi, Z and Liu, X and Wang, J and Guo, Y and Ren, D and Wu, H and Wang, X and Chen, B and Liu, Q}, title = {Identification and molecular characterization of Wolbachia strains in natural populations of Aedes albopictus in China.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {28}, pmid = {31937373}, issn = {1756-3305}, support = {Grant no. 81273139//National Natural Science Foundation of China (NSFC)/ ; Grant no. 2016YFC1200802//National Major Research and Development Program/ ; }, mesh = {Aedes/*microbiology ; Analysis of Variance ; Animals ; Bacterial Outer Membrane Proteins/genetics ; China/epidemiology ; Climate ; Cluster Analysis ; DNA, Bacterial/isolation & purification ; Dengue/epidemiology/prevention & control/*transmission ; Female ; Likelihood Functions ; Male ; Mosquito Vectors/*microbiology ; Multilocus Sequence Typing ; Phylogeny ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Aedes albopictus is naturally infected with Wolbachia spp., maternally transmitted bacteria that influence the reproduction of hosts. However, little is known regarding the prevalence of infection, multiple infection status, and the relationship between Wolbachia density and dengue outbreaks in different regions. Here, we assessed Wolbachia infection in natural populations of Ae. albopictus in China and compared Wolbachia density between regions with similar climates, without dengue and with either imported or local dengue.

RESULTS: To explore the prevalence of Wolbachia infection, Wolbachia DNA was detected in mosquito samples via PCR amplification of the 16S rRNA gene and the surface protein gene wsp. We found that 93.36% of Ae. albopictus in China were positive for Wolbachia. After sequencing gatB, coxA, hcpA, ftsZ, fbpA and wsp genes of Wolbachia strains, we identified a new sequence type (ST) of wAlbB (464/465). Phylogenetic analysis indicated that wAlbA and wAlbB strains formed a cluster with strains from other mosquitoes in a wsp-based maximum likelihood (ML) tree. However, in a ML tree based on multilocus sequence typing (MLST), wAlbB STs (464/465) did not form a cluster with Wolbachia strains from other mosquitoes. To better understand the association between Wolbachia spp. and dengue infection, the prevalence of Wolbachia in Ae. albopictus from different regions (containing local dengue cases, imported dengue cases and no dengue cases) was determined. We found that the prevalence of Wolbachia was lower in regions with only imported dengue cases.

CONCLUSIONS: The natural prevalence of Wolbachia infections in China was much lower than in other countries or regions. The phylogenetic relationships among Wolbachia spp. isolated from field-collected Ae. albopictus reflected the presence of dominant and stable strains. However, wAlbB (464/465) and Wolbachia strains did not form a clade with Wolbachia strains from other mosquitoes. Moreover, lower densities of Wolbachia in regions with only imported dengue cases suggest a relationship between fluctuations in Wolbachia density in field-collected Ae. albopictus and the potential for dengue invasion into these regions.}, } @article {pmid31927646, year = {2020}, author = {Bezerra-Santos, MA and Nogueira, BCF and Yamatogi, RS and Ramos, RAN and Galhardo, JA and Campos, AK}, title = {Ticks, fleas and endosymbionts in the ectoparasite fauna of the black-eared opossum Dipelphis aurita in Brazil.}, journal = {Experimental & applied acarology}, volume = {80}, number = {3}, pages = {329-338}, pmid = {31927646}, issn = {1572-9702}, mesh = {Anaplasmataceae/isolation & purification ; Animals ; Brazil ; Didelphis/*parasitology ; Flea Infestations/*veterinary ; Ixodidae/*microbiology ; Siphonaptera/*microbiology ; Tick Infestations/*veterinary ; Wolbachia/isolation & purification ; }, abstract = {Ticks and fleas are essential vectors of pathogens that affect humans and animals, and among their hosts, synanthropic animals such as the black-eared opossum, Didelphis aurita, play a role in public health due to their ability to move between urban centers and forested areas in Brazil. This study aimed to assess the ectoparasite fauna of D. aurita, as well as the presence of pathogens and endosymbionts in ticks and fleas. Opossums (n = 58) captured in Tomahawk livetraps were examined for ectoparasites, and their blood sampled for further analysis. Additionally, spleen samples were collected in individuals found dead. Samples were PCR screened for Rickettsia spp., Borrelia spp., Anaplasmataceae, and Babesia spp. Two tick species were morphologically identified as Ixodes loricatus 24/58 (41.4%) and Amblyomma sculptum 1/58 (1.7%). For fleas, Ctenocephalides felis was detected in 60.3% (35/58) of the animals, and Xenopsylla cheopis in 5.2% (3/58). PCR analysis detected Anaplasmataceae DNA in 34% (16/47) of pooled samples of C. felis, and in 66.7% (2/3) pooled samples of X. cheopis. Sequence analysis revealed Wolbachia pipientis symbiont in all positive samples. Tick, blood and spleen samples were all negative for the microorganisms assessed. These findings suggest that these arthropods circulate among wildlife and urban environments, which may implicate in their participation in the cycle of zoonotic pathogens among opossums, humans and companion animals.}, } @article {pmid31926932, year = {2020}, author = {Li, Y and Liu, X}, title = {Modeling and control of mosquito-borne diseases with Wolbachia and insecticides.}, journal = {Theoretical population biology}, volume = {132}, number = {}, pages = {82-91}, doi = {10.1016/j.tpb.2019.12.007}, pmid = {31926932}, issn = {1096-0325}, mesh = {*Aedes ; Animals ; Female ; Humans ; *Insecticides ; Male ; Mosquito Control ; *Vector Borne Diseases ; *Wolbachia ; }, abstract = {Mosquitoes cause more human suffering than any other organism. It is estimated that over one million people worldwide die from mosquito-borne diseases every year. With the continuous efforts of many researchers, Wolbachia gets more and more attention due to its characteristics of maternal transmission in mosquito population and it may cause cytoplasmic incompatibility (CI) which makes healthy females cannot fertilize normally after mating with infected males. In this paper, mathematical models are established to study Wolbachia transmission in mosquito population, and integrated mosquito control strategies are explored. Firstly, a classical ordinary differential system with general birth and death rate functions is established to describe the maternal transmission and CI effect. It is shown that the replacement strategy that the Wolbachia-uninfected mosquitoes are replaced by the infected ones is determined by the initial infection frequency. And Wolbachia spreads more easily for greater maternal transmission and CI rate. Moreover, all the wild mosquitoes will eventually be infected with Wolbachia if the maternal transmission is complete. Secondly, an impulsive state feedback control model is constructed to describe the integrated mosquito control. Besides Wolbachia, insecticides are sprayed when the quantity of mosquitoes reaches some Economic Threshold. The existence and stability of Wolbachia replacement periodic solution are discussed. Finally, some discussions are done and the future research directions are prospected.}, } @article {pmid31921019, year = {2019}, author = {Alfano, N and Tagliapietra, V and Rosso, F and Manica, M and Arnoldi, D and Pindo, M and Rizzoli, A}, title = {Changes in Microbiota Across Developmental Stages of Aedes koreicus, an Invasive Mosquito Vector in Europe: Indications for Microbiota-Based Control Strategies.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {2832}, pmid = {31921019}, issn = {1664-302X}, abstract = {Since it has been understood that gut microbiota of vector mosquitoes can influence their vector competence, efforts have been undertaken to develop new control strategies based on host microbiota manipulation, and aimed at suppressing the vector population or replacing it with a less competent one. For the proper design of such control strategies it is necessary to know the microbiota composition of the target vector species, how it is acquired, and how it changes throughout the host's life cycle. In this study, 16S rRNA amplicon high-throughput sequencing was used to characterize the changes in microbiota from the aquatic environment (larval breeding sites) to the different developmental stages of field-collected Aedes koreicus in Italy, an emerging invasive mosquito species in Europe and a potential vector of several pathogens. The bacterial communities of the aquatic breeding sites, larvae, pupae and adults showed distinctive structures to one another. Indeed, 84% of community members were unique to a given sample type. Nevertheless, almost 40% of the sequences generated were assigned to bacteria detected in all sample types, suggesting the importance of bacteria transstadially transmitted from water to the adult stage in constituting mosquito microbiota. Among these, genus C39 largely constituted water microbiota, family Burkholderiaceae was the most abundant in larvae and pupae, and genus Asaia dominated adult communities. In addition, Asaia constituted a core microbiota across all sample types. Our results suggest that the microbiota of Ae. koreicus mosquitoes is composed by a community which derives from the aquatic bacteria of the larval breeding sites, is then filtered by the larval gut, where only certain members are able to persist, rearranged by metamorphosis and finally modified by the change in diet at the adult stage. Understanding how the microbiota of Ae. koreicus changes through the mosquito life cycle represents a first step in selecting bacterial candidates for use in microbiota-based intervention measures for this species. The properties which Asaia exhibits in this species, such as dominance, high prevalence and transstadial transmission, prevent the use of Wolbachia but make Asaia an ideal candidate for paratransgenesis.}, } @article {pmid31919396, year = {2020}, author = {Garcia, GA and Hoffmann, AA and Maciel-de-Freitas, R and Villela, DAM}, title = {Aedes aegypti insecticide resistance underlies the success (and failure) of Wolbachia population replacement.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {63}, pmid = {31919396}, issn = {2045-2322}, mesh = {Aedes/*microbiology ; Animals ; *Insecticide Resistance/drug effects ; Insecticides/toxicity ; Models, Biological ; Population Dynamics ; Pyrethrins/toxicity ; Symbiosis ; Wolbachia/drug effects/isolation & purification/*physiology ; }, abstract = {Mosquitoes that carry Wolbachia endosymbionts may help control the spread of arboviral diseases, such as dengue, Zika and chikungunya. Wolbachia frequencies systematically increase only when the frequency-dependent advantage due to cytoplasmic incompatibility exceeds frequency-independent costs, which may be intrinsic to the Wolbachia and/or can be associated with the genetic background into which Wolbachia are introduced. Costs depend on field conditions such as the environmental pesticide load. Introduced mosquitoes need adequate protection against insecticides to ensure survival after release. We model how insecticide resistance of transinfected mosquitoes determines the success of local Wolbachia introductions and link our theoretical results to field data. Two Ae. aegypti laboratory strains carrying Wolbachia were released in an isolated district of Rio de Janeiro, Brazil: wMelBr (susceptible to pyrethroids) and wMelRio (resistant to pyrethroids). Our models elucidate why releases of the susceptible strain failed to result in Wolbachia establishment, while releases of the resistant strain led to Wolbachia transforming the native Ae. aegypti population. The results highlight the importance of matching insecticide resistance levels in release stocks to those in the target natural populations during Wolbachia deployment.}, } @article {pmid31912447, year = {2020}, author = {Reeves, DD and Price, SL and Ramalho, MO and Moreau, CS}, title = {The Diversity and Distribution of Wolbachia, Rhizobiales, and Ophiocordyceps Within the Widespread Neotropical Turtle Ant, Cephalotes atratus (Hymenoptera: Formicidae).}, journal = {Neotropical entomology}, volume = {49}, number = {1}, pages = {52-60}, pmid = {31912447}, issn = {1678-8052}, support = {NSF DBI 1559779//National Science Foundation Research Experience/ ; NSF DEB 1900357//National Science Foundation/ ; }, mesh = {Alphaproteobacteria/*genetics ; Animals ; Ants/*microbiology ; Genetic Variation ; Genotype ; Hypocreales/*genetics ; South America ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Ants are an ecologically and evolutionarily diverse group, and they harbor a wide range of symbiotic microbial communities that often greatly affect their biology. Turtle ants (genus Cephalotes) engage in mutualistic relationships with gut bacteria and are exploited by microbial parasites. Studies have shown that associations among these microbial lineages and the turtle ant hosts vary geographically. However, these studies have been limited, and thorough within-species analyses of the variation and structure of these microbial communities have yet to be conducted. The giant turtle ant, Cephalotes atratus (Linnaeus 1758), is a geographically widespread, genetically diverse Neotropical species that has been sampled extensively across its geographic range, making it ideal for analysis of microbial associations. In this study, we verified the presence, genetic variation, and geographic patterns at the individual, colony, and population level of three microbial groups associated with the giant turtle ant: Wolbachia, a genus of facultative bacteria which are often parasitic, affecting host reproduction; Rhizobiales, a mutualistic order of bacteria hypothesized to be an obligate nutritional symbiont in turtle ants; and Ophiocordyceps, a genus of endoparasitic fungi infecting many arthropod species by manipulating their behavior for fungal reproduction. In this study, we found varying degrees of prevalence for two distantly related genotypes (haplogroups) of Wolbachia and high degree of prevalence of Rhizobiales across colonies with little genetic variation. In addition, we found low occurrence of Ophiocordyceps. This study highlights a key first step in understanding the diversity, distribution, and prevalence of the microbial community of C. atratus.}, } @article {pmid31910374, year = {2020}, author = {Chrostek, E and Hurst, GDD and McGraw, EA}, title = {Infectious Diseases: Antiviral Wolbachia Limits Dengue in Malaysia.}, journal = {Current biology : CB}, volume = {30}, number = {1}, pages = {R30-R32}, doi = {10.1016/j.cub.2019.11.046}, pmid = {31910374}, issn = {1879-0445}, mesh = {*Aedes ; Animals ; *Antiviral Agents ; *Dengue/transmission ; *Dengue Virus ; Humans ; Malaysia ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Vector-borne viral diseases pose an urgent public health challenge, particularly in the tropics. Field releases of mosquitoes carrying bacterial symbionts that reduce vector competence are ongoing in Kuala Lumpur, Malaysia. Early results show that wAlbB Wolbachia can persist in mosquitoes in urban settings and decrease dengue incidence in humans.}, } @article {pmid31900308, year = {2020}, author = {Madhav, M and Parry, R and Morgan, JAT and James, P and Asgari, S}, title = {Wolbachia Endosymbiont of the Horn Fly (Haematobia irritans irritans): a Supergroup A Strain with Multiple Horizontally Acquired Cytoplasmic Incompatibility Genes.}, journal = {Applied and environmental microbiology}, volume = {86}, number = {6}, pages = {}, pmid = {31900308}, issn = {1098-5336}, mesh = {Animals ; Gene Transfer, Horizontal ; *Genes, Bacterial ; Muscidae/*microbiology ; *Symbiosis/genetics ; Wolbachia/genetics/*physiology ; }, abstract = {The horn fly, Haematobia irritansirritans, is a hematophagous parasite of livestock distributed throughout Europe, Africa, Asia, and the Americas. Welfare losses on livestock due to horn fly infestation are estimated to cost between $1 billion and $2.5 billion (U.S. dollars) annually in North America and Brazil. The endosymbiotic bacterium Wolbachia pipientis is a maternally inherited manipulator of reproductive biology in arthropods and naturally infects laboratory colonies of horn flies from Kerrville, TX, and Alberta, Canada, but it has also been identified in wild-caught samples from Canada, the United States, Mexico, and Hungary. Reassembly of PacBio long-read and Illumina genomic DNA libraries from the Kerrville H. i. irritans genome project allowed for a complete and circularized 1.3-Mb Wolbachia genome (wIrr). Annotation of wIrr yielded 1,249 coding genes, 34 tRNAs, 3 rRNAs, and 5 prophage regions. Comparative genomics and whole-genome Bayesian evolutionary analysis of wIrr compared to published Wolbachia genomes suggested that wIrr is most closely related to and diverged from Wolbachia supergroup A strains known to infect Drosophila spp. Whole-genome synteny analyses between wIrr and closely related genomes indicated that wIrr has undergone significant genome rearrangements while maintaining high nucleotide identity. Comparative analysis of the cytoplasmic incompatibility (CI) genes of wIrr suggested two phylogenetically distinct CI loci and acquisition of another cifB homolog from phylogenetically distant supergroup A Wolbachia strains, suggesting horizontal acquisition of these loci. The wIrr genome provides a resource for future examination of the impact Wolbachia may have in both biocontrol and potential insecticide resistance of horn flies.IMPORTANCE Horn flies, Haematobia irritans irritans, are obligate hematophagous parasites of cattle having significant effects on production and animal welfare. Control of horn flies mainly relies on the use of insecticides, but issues with resistance have increased interest in development of alternative means of control. Wolbachia pipientis is an endosymbiont bacterium known to have a range of effects on host reproduction, such as induction of cytoplasmic incompatibility, feminization, male killing, and also impacts vector transmission. These characteristics of Wolbachia have been exploited in biological control approaches for a range of insect pests. Here we report the assembly and annotation of the circular genome of the Wolbachia strain of the Kerrville, TX, horn fly (wIrr). Annotation of wIrr suggests its unique features, including the horizontal acquisition of additional transcriptionally active cytoplasmic incompatibility loci. This study provides the foundation for future studies of Wolbachia-induced biological effects for control of horn flies.}, } @article {pmid31874958, year = {2019}, author = {Lefoulon, E and Vaisman, N and Frydman, HM and Sun, L and Voland, L and Foster, JM and Slatko, BE}, title = {Author Correction: Large Enriched Fragment Targeted Sequencing (LEFT-SEQ) Applied to Capture of Wolbachia Genomes.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {20184}, doi = {10.1038/s41598-019-55305-5}, pmid = {31874958}, issn = {2045-2322}, abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.}, } @article {pmid31870298, year = {2019}, author = {Asimakis, ED and Doudoumis, V and Hadapad, AB and Hire, RS and Batargias, C and Niu, C and Khan, M and Bourtzis, K and Tsiamis, G}, title = {Detection and characterization of bacterial endosymbionts in Southeast Asian tephritid fruit fly populations.}, journal = {BMC microbiology}, volume = {19}, number = {Suppl 1}, pages = {290}, pmid = {31870298}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Gene Transfer, Horizontal ; Pest Control, Biological ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Symbiosis ; Tephritidae/*microbiology ; }, abstract = {BACKGROUND: Various endosymbiotic bacteria, including Wolbachia of the Alphaproteobacteria, infect a wide range of insects and are capable of inducing reproductive abnormalities to their hosts such as cytoplasmic incompatibility (CI), parthenogenesis, feminization and male-killing. These extended phenotypes can be potentially exploited in enhancing environmentally friendly methods, such as the sterile insect technique (SIT), for controlling natural populations of agricultural pests. The goal of the present study is to investigate the presence of Wolbachia, Spiroplasma, Arsenophonus and Cardinium among Bactrocera, Dacus and Zeugodacus flies of Southeast Asian populations, and to genotype any detected Wolbachia strains.

RESULTS: A specific 16S rRNA PCR assay was used to investigate the presence of reproductive parasites in natural populations of nine different tephritid species originating from three Asian countries, Bangladesh, China and India. Wolbachia infections were identified in Bactrocera dorsalis, B. correcta, B. scutellaris and B. zonata, with 12.2-42.9% occurrence, Entomoplasmatales in B. dorsalis, B. correcta, B. scutellaris, B. zonata, Zeugodacus cucurbitae and Z. tau (0.8-14.3%) and Cardinium in B. dorsalis and Z. tau (0.9-5.8%), while none of the species tested, harbored infections with Arsenophonus. Infected populations showed a medium (between 10 and 90%) or low (< 10%) prevalence, ranging from 3 to 80% for Wolbachia, 2 to 33% for Entomoplasmatales and 5 to 45% for Cardinium. Wolbachia and Entomoplasmatales infections were found both in tropical and subtropical populations, the former mostly in India and the latter in various regions of India and Bangladesh. Cardinium infections were identified in both countries but only in subtropical populations. Phylogenetic analysis revealed the presence of Wolbachia with some strains belonging either to supergroup B or supergroup A. Sequence analysis revealed deletions of variable length and nucleotide variation in three Wolbachia genes. Spiroplasma strains were characterized as citri-chrysopicola-mirum and ixodetis strains while the remaining Entomoplasmatales to the Mycoides-Entomoplasmataceae clade. Cardinium strains were characterized as group A, similar to strains infecting Encarsia pergandiella.

CONCLUSIONS: Our results indicated that in the Southeast natural populations examined, supergroup A Wolbachia strain infections were the most common, followed by Entomoplasmatales and Cardinium. In terms of diversity, most strains of each bacterial genus detected clustered in a common group. Interestingly, the deletions detected in three Wolbachia genes were either new or similar to those of previously identified pseudogenes that were integrated in the host genome indicating putative horizontal gene transfer events in B. dorsalis, B. correcta and B. zonata.}, } @article {pmid31870290, year = {2019}, author = {Conte, CA and Segura, DF and Milla, FH and Augustinos, A and Cladera, JL and Bourtzis, K and Lanzavecchia, SB}, title = {Wolbachia infection in Argentinean populations of Anastrepha fraterculus sp1: preliminary evidence of sex ratio distortion by one of two strains.}, journal = {BMC microbiology}, volume = {19}, number = {Suppl 1}, pages = {289}, pmid = {31870290}, issn = {1471-2180}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Female ; Male ; Multilocus Sequence Typing ; Phylogeny ; Sex Ratio ; Sexual Behavior, Animal ; Tephritidae/*microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Wolbachia, one of the most abundant taxa of intracellular Alphaproteobacteria, is widespread among arthropods and filarial nematodes. The presence of these maternally inherited bacteria is associated with modifications of host fitness, including a variety of reproductive abnormalities, such as cytoplasmic incompatibility, thelytokous parthenogenesis, host feminization and male-killing. Wolbachia has attracted much interest for its role in biological, ecological and evolutionary processes as well as for its potential use in novel and environmentally-friendly strategies for the control of insect pests and disease vectors including a major agricultural pest, the South American fruit fly, Anastrepha fraterculus Wiedemann (Diptera: Tephritidae).

RESULTS: We used wsp, 16S rRNA and a multilocus sequence typing (MLST) scheme including gatB, coxA, hcpA, fbpA, and ftsZ genes to detect and characterize the Wolbachia infection in laboratory strains and wild populations of A. fraterculus from Argentina. Wolbachia was found in all A. fraterculus individuals studied. Nucleotide sequences analysis of wsp gene allowed the identification of two Wolbachia nucleotide variants (named wAfraCast1_A and wAfraCast2_A). After the analysis of 76 individuals, a high prevalence of the wAfraCast2_A variant was found both, in laboratory (82%) and wild populations (95%). MLST analysis identified both Wolbachia genetic variants as sequence type 13. Phylogenetic analysis of concatenated MLST datasets clustered wAfraCast1/2_A in the supergroup A. Paired-crossing experiments among single infected laboratory strains showed a phenotype specifically associated to wAfraCast1_A that includes slight detrimental effects on larval survival, a female-biased sex ratio; suggesting the induction of male-killing phenomena, and a decreased proportion of females producing descendants that appears attributable to the lack of sperm in their spermathecae.

CONCLUSIONS: We detected and characterized at the molecular level two wsp gene sequence variants of Wolbachia both in laboratory and wild populations of A. fraterculus sp.1 from Argentina. Crossing experiments on singly-infected A. fraterculus strains showed evidence of a male killing-like mechanism potentially associated to the wAfraCast1_A - A. fraterculus interactions. Further mating experiments including antibiotic treatments and the analysis of early and late immature stages of descendants will contribute to our understanding of the phenotypes elicited by the Wolbachia variant wAfraCast1_A in A. fraterculus sp.1.}, } @article {pmid31869759, year = {2020}, author = {Hübner, MP and Gunderson, E and Vogel, I and Bulman, CA and Lim, KC and Koschel, M and Ehrens, A and Frohberger, SJ and Fendler, M and Tricoche, N and Voronin, D and Steven, A and Chi, V and Bakowski, MA and Woods, AK and Petrassi, HM and McNamara, CW and Beerntsen, B and Chappell, L and Sullivan, W and Taylor, MJ and Turner, JD and Hoerauf, A and Lustigman, S and Sakanari, JA}, title = {Short-course quinazoline drug treatments are effective in the Litomosoides sigmodontis and Brugia pahangi jird models.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {12}, number = {}, pages = {18-27}, pmid = {31869759}, issn = {2211-3207}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/*therapeutic use ; Brugia pahangi/drug effects ; Female ; Filariasis/*drug therapy/microbiology ; Filarioidea/drug effects ; Gerbillinae/microbiology/parasitology ; Microfilariae/drug effects ; Onchocerciasis/*drug therapy ; Quinazolines/administration & dosage/*therapeutic use ; Symbiosis/drug effects ; Wolbachia/*drug effects ; }, abstract = {The quinazolines CBR417 and CBR490 were previously shown to be potent anti-wolbachials that deplete Wolbachia endosymbionts of filarial nematodes and present promising pre-clinical candidates for human filarial diseases such as onchocerciasis. In the present study we tested both candidates in two models of chronic filarial infection, namely the Litomosoides sigmodontis and Brugia pahangi jird model and assessed their long-term effect on Wolbachia depletion, microfilariae counts and filarial embryogenesis 16-18 weeks after treatment initiation (wpt). Once per day (QD) oral treatment with CBR417 (50 mg/kg) for 4 days or twice per day (BID) with CBR490 (25 mg/kg) for 7 days during patent L. sigmodontis infection reduced the Wolbachia load by >99% and completely cleared peripheral microfilaremia from 10-14 wpt. Similarly, 7 days of QD treatments (40 mg/kg) with CBR417 or CBR490 cleared >99% of Wolbachia from B. pahangi and reduced peritoneal microfilariae counts by 93% in the case of CBR417 treatment. Transmission electron microscopy analysis indicated intensive damage to the B. pahangi ovaries following CBR417 treatment and in accordance filarial embryogenesis was inhibited in both models after CBR417 or CBR490 treatment. Suboptimal treatment regimens of CBR417 or CBR490 did not lead to a maintained reduction of the microfilariae and Wolbachia load. In conclusion, CBR417 or CBR490 are pre-clinical candidates for filarial diseases, which achieve long-term clearance of Wolbachia endosymbionts of filarial nematodes, inhibit filarial embryogenesis and clear microfilaremia with treatments as short as 7 days.}, } @article {pmid31863009, year = {2019}, author = {Fauver, JR and Martin, J and Weil, GJ and Mitreva, M and Fischer, PU}, title = {De novo Assembly of the Brugia malayi Genome Using Long Reads from a Single MinION Flowcell.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {19521}, pmid = {31863009}, issn = {2045-2322}, support = {R01 AI081803/AI/NIAID NIH HHS/United States ; R01 GM097435/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*genetics ; Female ; Genome, Mitochondrial/genetics ; Genome, Protozoan/genetics ; Sequence Analysis, DNA ; }, abstract = {Filarial nematode infections cause a substantial global disease burden. Genomic studies of filarial worms can improve our understanding of their biology and epidemiology. However, genomic information from field isolates is limited and available reference genomes are often discontinuous. Single molecule sequencing technologies can reduce the cost of genome sequencing and long reads produced from these devices can improve the contiguity and completeness of genome assemblies. In addition, these new technologies can make generation and analysis of large numbers of field isolates feasible. In this study, we assessed the performance of the Oxford Nanopore Technologies MinION for sequencing and assembling the genome of Brugia malayi, a human parasite widely used in filariasis research. Using data from a single MinION flowcell, a 90.3 Mb nuclear genome was assembled into 202 contigs with an N50 of 2.4 Mb. This assembly covered 96.9% of the well-defined B. malayi reference genome with 99.2% identity. The complete mitochondrial genome was obtained with individual reads and the nearly complete genome of the endosymbiotic bacteria Wolbachia was assembled alongside the nuclear genome. Long-read data from the MinION produced an assembly that approached the quality of a well-established reference genome using comparably fewer resources.}, } @article {pmid31847836, year = {2019}, author = {Kyritsis, GA and Augustinos, AA and Livadaras, I and Cáceres, C and Bourtzis, K and Papadopoulos, NT}, title = {Medfly-Wolbachia symbiosis: genotype x genotype interactions determine host's life history traits under mass rearing conditions.}, journal = {BMC biotechnology}, volume = {19}, number = {Suppl 2}, pages = {96}, pmid = {31847836}, issn = {1472-6750}, mesh = {Anaplasmataceae Infections/*veterinary ; Animals ; Ceratitis capitata/classification/genetics/microbiology/*physiology ; Female ; Fertility ; Genotype ; Male ; Sexual Behavior, Animal ; Symbiosis ; Wolbachia/classification/genetics/*pathogenicity ; }, abstract = {BACKGROUND: Wolbachia pipientis is a widespread, obligatory intracellular and maternally inherited bacterium, that induces a wide range of reproductive alterations to its hosts. Cytoplasmic Incompatibility (CI) is causing embryonic lethality, the most common of them. Despite that Wolbachia-borne sterility has been proposed as an environmental friendly pest control method (Incompatible Insect Technique, IIT) since 1970s, the fact that Wolbachia modifies important fitness components of its hosts sets severe barriers to IIT implementation. Mass rearing of Mediterranean fruit fly, Ceratitis capitata (medfly), is highly optimized given that this pest is a model species regarding the implementation of another sterility based pest control method, the Sterile Insect Technique (SIT). We used the medfly-Wolbachia symbiotic association, as a model system, to study the effect of two different Wolbachia strains, on the life history traits of 2 C. capitata lines with different genomic background.

RESULTS: Wolbachia effects are regulated by both C. capitata genetic background and the Wolbachia strain. Wolbachia infection reduces fertility rates in both C. capitata genetic backgrounds and shortens the pre-pupa developmental duration in the GSS strain. On the other hand, regardless of the strain of Wolbachia (wCer2, wCer4) infection does not affect either the sex ratio or the longevity of adults. wCer4 infection imposed a reduction in females' fecundity but wCer2 did not. Male mating competitiveness, adults flight ability and longevity under water and food deprivation were affected by both the genetic background of medfly and the strain of Wolbachia (genotype by genotype interaction).

CONCLUSION: Wolbachia infection could alter important life history traits of mass-reared C. capitata lines and therefore the response of each genotype on the Wolbachia infection should be considered toward ensuring the productivity of the Wolbachia-infected insects under mass-rearing conditions.}, } @article {pmid31847764, year = {2019}, author = {Wedell, N and Price, TAR and Lindholm, AK}, title = {Gene drive: progress and prospects.}, journal = {Proceedings. Biological sciences}, volume = {286}, number = {1917}, pages = {20192709}, pmid = {31847764}, issn = {1471-2954}, mesh = {Aedes ; Animals ; Biological Evolution ; Culicidae ; Dengue ; *Disease Vectors ; *Gene Drive Technology ; Mosquito Vectors ; Reproduction ; Wolbachia ; Zika Virus ; Zika Virus Infection ; }, abstract = {Gene drive is a naturally occurring phenomenon in which selfish genetic elements manipulate gametogenesis and reproduction to increase their own transmission to the next generation. Currently, there is great excitement about the potential of harnessing such systems to control major pest and vector populations. If synthetic gene drive systems can be constructed and applied to key species, they may be able to rapidly spread either modifying or eliminating the targeted populations. This approach has been lauded as a revolutionary and efficient mechanism to control insect-borne diseases and crop pests. Driving endosymbionts have already been deployed to combat the transmission of dengue and Zika virus in mosquitoes. However, there are a variety of barriers to successfully implementing gene drive techniques in wild populations. There is a risk that targeted organisms will rapidly evolve an ability to suppress the synthetic drive system, rendering it ineffective. There are also potential risks of synthetic gene drivers invading non-target species or populations. This Special Feature covers the current state of affairs regarding both natural and synthetic gene drive systems with the aim to identify knowledge gaps. By understanding how natural drive systems spread through populations, we may be able to better predict the outcomes of synthetic drive release.}, } @article {pmid31846415, year = {2020}, author = {Bishop, C and Parry, R and Asgari, S}, title = {Effect of Wolbachia wAlbB on a positive-sense RNA negev-like virus: a novel virus persistently infecting Aedes albopictus mosquitoes and cells.}, journal = {The Journal of general virology}, volume = {101}, number = {2}, pages = {216-225}, doi = {10.1099/jgv.0.001361}, pmid = {31846415}, issn = {1465-2099}, mesh = {Aedes/*virology ; Animals ; Cell Line ; *Coinfection/microbiology/virology ; Genome, Viral ; *Insect Viruses/classification/genetics/growth & development/isolation & purification ; Microbial Interactions ; Phylogeny ; *RNA Viruses/classification/genetics/growth & development/isolation & purification ; *Wolbachia ; }, abstract = {The Aedes aegypti mosquito is the primary vector of several medically important arboviruses. The endosymbiotic bacterium, Wolbachia pipientis, has emerged as a means of blocking transmission of arboviruses such as dengue and Zika viruses. One Wolbachia strain that has shown potential in field trials is wAlbB, a naturally occurring Wolbachia strain of the Asian tiger mosquito Aedes albopictus. When transinfected into Ae. aegypti, wAlbB exhibits strong virus inhibition. In addition to modulating arboviruses, Wolbachia also modulates some insect-specific viruses. Here, we explored the effect of Wolbachia on the virome of the Ae. albopictus cell line Aa23 naturally infected with wAlbB and also a stably transinfected recipient Ae. aegypti cell line (Aag2.wAlbB). RNA sequencing and bioinformatic analysis on both cell lines revealed an 11 kb genome of a single-stranded positive-sense RNA negev-like virus related to the recently proposed negevirus taxon. We denoted this novel virus as Aedes albopictus negev-like virus (AalNLV). Tetracycline clearance of Wolbachia from Aa23 cells did not significantly affect AalNLV levels, while in Aag2.wAlbB cells, a significant increase in virus genome RNA copies was observed. We further investigated the inhibitory effect of wAlbB on AalNLV and another positive-sense RNA virus, cell fusing agent virus, which is present in Aag2 cells and known to be suppressed by Wolbachia. wAlbB suppressed both viruses, with the effect on AalNLV being more striking. The findings from this study further supplement our understanding of the complex interaction between Wolbachia, host and virome.}, } @article {pmid31843539, year = {2020}, author = {Li, C and He, M and Yun, Y and Peng, Y}, title = {Co-infection with Wolbachia and Cardinium may promote the synthesis of fat and free amino acids in a small spider, Hylyphantes graminicola.}, journal = {Journal of invertebrate pathology}, volume = {169}, number = {}, pages = {107307}, doi = {10.1016/j.jip.2019.107307}, pmid = {31843539}, issn = {1096-0805}, mesh = {Amino Acids/*metabolism ; Animals ; Bacteroidetes/*physiology ; Fats/*metabolism ; Spiders/*metabolism/microbiology ; Symbiosis ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {Associations between endosymbiotic bacteria and their hosts are widespread in nature and have been demonstrated extensively; however, only a few studies have examined how facultative symbionts affect host nutrition and metabolism. To gain insight into the associations between facultative symbionts and host nutrition and metabolic activity, we detected endosymbiotic infection in a small spider species, Hylyphantes graminicola, and established two infectious strains, i.e., W[-]C[+] (Wolbachia negative, Cardinium positive) and W[+]C[+] (Wolbachia positive, Cardinium positive). We then determined the content of fat and free amino acids in W[-]C[+] and W[+]C[+] spiders, respectively. We also detected the transcriptome of H. graminicola and the expression of genes involved in fat and amino acid metabolism at different host ages. Results showed that fat content in W[+]C[+] spiders was higher than that in W[-]C[+] spiders, and free amino acid content was higher in W[+]C[+] males than W[-]C[+] males, with no difference observed in females. Transcriptome analysis identified 144 (W[-]C[+] vs W[+]C[+]) differentially expressed genes (DEGs). Moreover, the expression of five genes involved in fat and amino acid metabolism were significantly up-regulated in the third, fourth, and fifth instar stages in W[+]C[+] spiders. This study indicated that Wolbachia and Cardinium co-infection had a pivotal effect on fat and amino acid synthesis in hosts. Moreover, our results provide strong evidence explaining the long-term coexistence of hosts and endosymbionts.}, } @article {pmid31840281, year = {2020}, author = {Duan, DY and Zhou, HM and Cheng, TY}, title = {Comparative analysis of microbial community in the whole body and midgut from fully engorged and unfed female adult Melophagus ovinus.}, journal = {Medical and veterinary entomology}, volume = {34}, number = {2}, pages = {215-224}, doi = {10.1111/mve.12424}, pmid = {31840281}, issn = {1365-2915}, mesh = {Animals ; Bacteria/*isolation & purification ; China ; DNA, Bacterial/analysis ; Diptera/*microbiology ; Feeding Behavior ; Female ; Gastrointestinal Microbiome ; *Microbiota ; RNA, Ribosomal, 16S/analysis ; }, abstract = {Melophagus ovinus is a type of ectoparasite infesting sheep. Data regarding the comprehensive bacterial community associated with the whole body and midgut of M. ovinus under different engorged statuses are required. Melophagus ovinus were collected from the city of Jiuquan, China. Bacterial DNA was extracted from the whole body and midgut of fully engorged female adults, or newly hatched and unfed adult female M. ovinus. The 16S rRNA gene V3-V4 hypervariable regions were sequenced using the IonS5™XL platform (Thermo Fisher Scientific, Waltham, MA, U.S.A.). The whole body bacterial diversity of the newly hatched, unfed adult females was greater compared with that of the other three samples. Proteobacteria was the dominant bacterial phylum in all of the samples. Of the 42 total bacterial genera present in all of the experimental samples, Arsenophonus, Bartonella and Wolbachia were the dominant genera. The relative abundance of Arsenophonus in midgut was greater than that in the whole body. The relative abundance of Bartonella in fully engorged adults was far greater than those in newly hatched, unfed adults. The relative abundance of Wolbachia was highest in the whole body of newly hatched, unfed adults. Seventeen bacterial species were identified in all experimental samples. Bartonella chomelii, Streptococcus hyointestinalis and Escherichia coli were the first species reported in M. ovinus.}, } @article {pmid31828225, year = {2019}, author = {Sawasdichai, S and Chaumeau, V and Dah, T and Kulabkeeree, T and Kajeechiwa, L and Phanaphadungtham, M and Trakoolchengkaew, M and Kittiphanakun, P and Akararungrot, Y and Oo, K and Delmas, G and White, NJ and Nosten, FH}, title = {Detection of diverse Wolbachia 16S rRNA sequences at low titers from malaria vectors in Kayin state, Myanmar.}, journal = {Wellcome open research}, volume = {4}, number = {}, pages = {11}, pmid = {31828225}, issn = {2398-502X}, support = {/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Background : Natural Wolbachia infections in malaria mosquitoes were recently reported in Africa, and negatively correlated with the development of Plasmodium falciparum in the vectors. The occurrence and effects of Wolbachia infections outside Africa have not been described and may have been underestimated. Methods : Mosquitoes were collected by human-landing catch during May and June 2017 in ten villages in Kayin state, Myanmar. Closely related species of malaria vectors were identified with molecular assays. 16S rRNA Wolbachia DNA sequences were detected with quantitative real-time PCR. Results: Low titer of Wolbachia DNA was detected in 13/370 samples in six malaria vector species. Sequences were diverse and different from those described in the African malaria mosquitoes. Conclusion: The detection of Wolbachia DNA in malaria mosquitoes from Kayin state warrants further investigations to understand better the ecology and biology of Anopheles- Wolbachia interactions in Southeast Asia.}, } @article {pmid31822714, year = {2019}, author = {Li, K and Stanojević, M and Stamenković, G and Ilić, B and Paunović, M and Lu, M and Pešić, B and Đurić Maslovara, I and Siljic, M and Cirkovic, V and Zhang, Y}, title = {Insight into diversity of bacteria belonging to the order Rickettsiales in 9 arthropods species collected in Serbia.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {18680}, pmid = {31822714}, issn = {2045-2322}, mesh = {Animals ; *Biodiversity ; Biological Evolution ; DNA, Bacterial/genetics ; *Genetic Variation ; Ixodes/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsiales/*classification/isolation & purification ; Seasons ; Serbia ; }, abstract = {Rickettsiales bacteria in arthropods play a significant role in both public health and arthropod ecology. However, the extensive genetic diversity of Rickettsiales endosymbionts of arthropods is still to be discovered. In 2016, 515 arthropods belonging to 9 species of four classes (Insecta, Chilopoda, Diplopoda and Arachnida) were collected in Serbia. The presence and genetic diversity of Rickettsiales bacteria were evaluated by characterizing the 16S rRNA (rrs), citrate synthase (gltA) and heat shock protein (groEL) genes. The presence of various Rickettsiales bacteria was identified in the majority of tested arthropod species. The results revealed co-circulation of five recognized Rickettsiales species including Rickettsia, Ehrlichia and Wolbachia, as well as four tentative novel species, including one tentative novel genus named Neowolbachia. These results suggest the remarkable genetic diversity of Rickettsiales bacteria in certain arthropod species in this region. Furthermore, the high prevalence of spotted fever group Rickettsia in Ixodes ricinus ticks highlights the potential public health risk of human Rickettsia infection.}, } @article {pmid31820073, year = {2020}, author = {Arai, H and Lin, SR and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Closely Related Male-Killing and Nonmale-Killing Wolbachia Strains in the Oriental Tea Tortrix Homona magnanima.}, journal = {Microbial ecology}, volume = {79}, number = {4}, pages = {1011-1020}, doi = {10.1007/s00248-019-01469-6}, pmid = {31820073}, issn = {1432-184X}, mesh = {Animals ; Insect Proteins/analysis ; Larva/genetics/growth & development/microbiology ; Moths/genetics/growth & development/*microbiology ; Phenotype ; Sex Factors ; Sex Ratio ; Symbiosis ; Taiwan ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are inherited intracellular bacteria that cause male-specific death in some arthropods, called male-killing. To date, three Wolbachia strains have been identified in the oriental tea tortrix Homona magnanima (Tortricidae, Lepidoptera); however, none of these caused male-killing in the Japanese population. Here, we describe a male-killing Wolbachia strain in Taiwanese H. magnanima. From field-collected H. magnanima, two female-biased host lines were established, and antibiotic treatments revealed Wolbachia (wHm-t) as the causative agent of male-killing. The wsp and MLST genes in wHm-t are identical to corresponding genes in the nonmale-killing strain wHm-c from the Japanese population, implying a close relationship of the two strains. Crossing the Japanese and Taiwanese H. magnanima revealed that Wolbachia genotype rather than the host genetic background was responsible for the presence of the male-killing phenotype. Quantitative PCR analyses revealed that the density of wHm-t was higher than that of other Wolbachia strains in H. magnanima, including wHm-c. The densities of wHm-t were also heterogeneous between host lines. Notably, wHm-t in the low-density and high-density lines carried identical wsp and MLST genes but had distinct lethal patterns. Furthermore, over 90% of field-collected lines of H. magnanima in Taiwan were infected with wHm-t, although not all host lines harboring wHm-t showed male-killing. The host lines that showed male-killing harbored a high density of Wolbachia compared to the host lines that did not show male-killing. Thus, the differences in the phenotypes appear to be dependent on biological and genetic characteristics of closely related Wolbachia strains.}, } @article {pmid31811033, year = {2020}, author = {Li, TP and Zhou, CY and Zha, SS and Gong, JT and Xi, Z and Hoffmann, AA and Hong, XY}, title = {Stable Establishment of Cardinium spp. in the Brown Planthopper Nilaparvata lugens despite Decreased Host Fitness.}, journal = {Applied and environmental microbiology}, volume = {86}, number = {4}, pages = {}, pmid = {31811033}, issn = {1098-5336}, mesh = {Animals ; Cytophagaceae/*physiology ; Female ; Genetic Fitness ; Hemiptera/growth & development/*microbiology ; Male ; Nymph/growth & development/microbiology ; *Symbiosis ; }, abstract = {The brown planthopper Nilaparvata lugens (Hemiptera) is a major pest of rice crops in Asia. Artificial transinfections of Wolbachia have recently been used for reducing host impacts, but transinfections have not yet been undertaken with another important endosymbiont, Cardinium This endosymbiont can manipulate the reproduction of hosts through phenotypes such as cytoplasmic incompatibility (CI), which is strong in the related white-backed planthopper, Sogatella furcifera (Hemiptera). Here, we stably infected N. lugens with Cardinium from S. furcifera and showed that it exhibits perfect maternal transmission in N. lugens The density of Cardinium varied across developmental stages and tissues of the transinfected host. Cardinium did not induce strong CI in N. lugens, likely due to its low density in testicles. The infection did decrease fecundity and hatching rate in the transinfected host, but a decrease in fecundity was not apparent when transinfected females mated with Wolbachia-infected males. The experiments show the feasibility of transferring Cardinium endosymbionts across hosts, but the deleterious effects of Cardinium on N. lugens limit its potential to spread in wild populations of N. lugens in the absence of strong CI.IMPORTANCE In this study we established a Cardinium-infected N. lugens line that possessed complete maternal transmission. Cardinium had a widespread distribution in tissues of N. lugens, and this infection decreased the fecundity and hatching rate of the host. Our findings emphasize the feasibility of transinfection of Cardinium in insects, which expands the range of endosymbionts that could be manipulated for pest control.}, } @article {pmid31805300, year = {2020}, author = {Liu, Y and Fan, ZY and An, X and Shi, PQ and Ahmed, MZ and Qiu, BL}, title = {A single-pair method to screen Rickettsia-infected and uninfected whitefly Bemisia tabaci populations.}, journal = {Journal of microbiological methods}, volume = {168}, number = {}, pages = {105797}, doi = {10.1016/j.mimet.2019.105797}, pmid = {31805300}, issn = {1872-8359}, mesh = {Animals ; Female ; Hemiptera/*microbiology/physiology ; Male ; Microbiological Techniques/*methods ; *Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*genetics/physiology ; Symbiosis/*genetics ; }, abstract = {Bacterial endosymbionts such as Rickettsia and Wolbachia play prominent roles in the development and behaviour of their insect hosts, such as whiteflies, aphids, psyllids and mealybugs. Accumulating studies have emphasized the importance of establishing experimental insect populations that are either lacking or bearing certain species of endosymbionts, because they are the basis in which to reveal the biological role of individual symbionts. In this study, using Rickettsia as an example, we explored a "single-pair screening" method to establish Rickettsia infected and uninfected populations of whitefly Bemisia tabaci MEAM1 for further experimental use. The original host population had a relatively low infection rate of Rickettsia (< 35%). When B. tabaci adults newly emerged, unmated males and females were randomly selected, and released into a leaf cage that covered a healthy plant leaf in order to oviposit F1 generation eggs. Following 6 days of oviposition, the parents were recaptured and used for PCR detection. The F1 progeny, for which parents were either Rickettsia positive or negative, were used to produce the F2 generation, and similarly in turn for the F3, F4 and F5 generations respectively; if the infection status of Rickettsia was consistent in the F1 to F5 generations, then the populations can be used as Rickettsia positive or negative lines for further experiments. In addition, our phylogenetic analyses revealed that Rickettsia has high fidelity during the maternal transmission in different generations.}, } @article {pmid31801631, year = {2019}, author = {Wanji, S and Nji, TM and Hamill, L and Dean, L and Ozano, K and Njouendou, AJ and Abong, RA and Obie, ED and Amuam, A and Ekanya, R and Ndongmo, WPC and Ndzeshang, BL and Fung, EG and Nnamdi, DB and Nkimbeng, DA and Teghen, S and Kah, E and Piotrowski, H and Forrer, A and Khan, JAM and Woode, ME and Niessen, L and Watson, V and Njoumemi, Z and Murdoch, ME and Thomson, R and Theobald, S and Enyong, P and Turner, JD and Taylor, MJ}, title = {Implementation of test-and-treat with doxycycline and temephos ground larviciding as alternative strategies for accelerating onchocerciasis elimination in an area of loiasis co-endemicity: the COUNTDOWN consortium multi-disciplinary study protocol.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {574}, pmid = {31801631}, issn = {1756-3305}, mesh = {Animals ; Anthelmintics/*therapeutic use ; Cameroon ; Disease Eradication/*methods/organization & administration ; Doxycycline/*therapeutic use ; Feasibility Studies ; Health Plan Implementation ; Humans ; *Insecticides ; Ivermectin/therapeutic use ; Loiasis/epidemiology ; Onchocerca/drug effects ; Onchocerciasis/diagnosis/*drug therapy/prevention & control ; Patient Acceptance of Health Care ; Prevalence ; Public Health/methods ; Simuliidae/*parasitology ; *Temefos ; World Health Organization ; }, abstract = {BACKGROUND: Onchocerciasis is a priority neglected tropical disease targeted for elimination by 2025. The standard strategy to combat onchocerciasis is annual Community-Directed Treatment with ivermectin (CDTi). Yet, high prevalence rates and transmission persist following > 12 rounds in South-West Cameroon. Challenges include programme coverage, adherence to, and acceptability of ivermectin in an area of Loa loa co-endemicity. Loiasis patients harbouring heavy infections are at risk of potentially fatal serious adverse events following CDTi. Alternative strategies are therefore needed to achieve onchocerciasis elimination where CDTi effectiveness is suboptimal.

METHODS/DESIGN: We designed an implementation study to evaluate integrating World Health Organisation-endorsed alternative strategies for the elimination of onchocerciasis, namely test-and-treat with the macrofilaricide, doxycycline (TTd), and ground larviciding for suppression of blackfly vectors with the organophosphate temephos. A community-based controlled before-after intervention study will be conducted among > 2000 participants in 20 intervention (Meme River Basin) and 10 control (Indian River Basin) communities. The primary outcome measure is O. volvulus prevalence at follow-up 18-months post-treatment. The study involves four inter-disciplinary components: parasitology, entomology, applied social sciences and health economics. Onchocerciasis skin infection will be diagnosed by skin biopsy and Loa loa infection will be diagnosed by parasitological examination of finger-prick blood samples. A simultaneous clinical skin disease assessment will be made. Eligible skin-snip-positive individuals will be offered directly-observed treatment for 5 weeks with 100 mg/day doxycycline. Transmission assessments of onchocerciasis in the communities will be collected post-human landing catch of the local biting blackfly vector prior to ground larviciding with temephos every week (0.3 l/m[3]) until biting rate falls below 5/person/day. Qualitative research, including in-depth interviews and focus-group discussions will be used to assess acceptability and feasibility of the implemented alternative strategies among intervention recipients and providers. Health economics will assess the cost-effectiveness of the implemented interventions.

CONCLUSIONS: Using a multidisciplinary approach, we aim to assess the effectiveness of TTd, alone or in combination with ground larviciding, following a single intervention round and scrutinise the acceptability and feasibility of implementing at scale in similar hotspots of onchocerciasis infection, to accelerate onchocerciasis elimination.}, } @article {pmid31799614, year = {2020}, author = {Staunton, KM and Rohde, BB and Townsend, M and Liu, J and Desnoyer, M and Howell, P and Amos, B and Crawford, J and Snoad, N and Ritchie, SA}, title = {Investigating Male Aedes aegypti (Diptera: Culicidae) Attraction to Different Oviposition Containers Using Various Configurations of the Sound Gravid Aedes Trap.}, journal = {Journal of medical entomology}, volume = {57}, number = {3}, pages = {957-961}, doi = {10.1093/jme/tjz229}, pmid = {31799614}, issn = {1938-2928}, mesh = {Aedes/*physiology ; Animals ; *Chemotaxis ; Male ; *Mosquito Control ; *Odorants ; Queensland ; }, abstract = {Aedes aegypti (Linnaeus), the primary vectors of the arboviruses dengue virus and Zika virus, continue to expand their global distributions. In efforts to better control such species, several mosquito control programs are investigating the efficacy of rearing and releasing millions of altered male Aedes throughout landscapes to reduce populations and disease transmission risk. Unfortunately, little is known about Ae. aegypti, especially male, dispersal behaviors within urban habitats. We deployed Sound-producing Gravid Aedes Traps (SGATs) in Cairns, northern Australia, to investigate male Ae. aegypti attraction to various oviposition container configurations. The traps were arranged to include: 1) water only, 2) organically infused water, 3) infused water and L3 larvae, 4) infused water and a human-scented lure, and lastly 5) no water or olfactory attractant (dry). Our data suggest that males were more attracted to SGATs representing active larval sites than potential larval sites, but were equally attracted to dry SGATs relative to those containing water and/or infusion. Additionally, we found that female Ae. aegypti were equally attracted to wet SGATs, with or without infusion, but not dry ones. These results suggest that male Ae. aegypti within northern Australia are more attracted to active larval sites and equally attracted to dry containers as wet or infused ones. Additionally, female Ae. aegypti are unlikely to enter dry containers. Such findings contribute to our understanding of potentially attractive features for local and released Ae. aegypti throughout the northern Australian urban landscape.}, } @article {pmid31797888, year = {2019}, author = {Devescovi, F and Conte, CA and Augustinos, A and Martinez, EIC and Segura, DF and Caceres, C and Lanzavecchia, SB and Bourtzis, K}, title = {Symbionts do not affect the mating incompatibility between the Brazilian-1 and Peruvian morphotypes of the Anastrepha fraterculus cryptic species complex.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {18319}, pmid = {31797888}, issn = {2045-2322}, mesh = {Animals ; Embryonic Development ; Female ; Genetic Speciation ; Male ; Reproduction ; *Reproductive Isolation ; *Symbiosis ; *Tephritidae/microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {The South American fruit fly, Anastrepha fraterculus, is clearly undergoing a speciation process. Among others, two of their morphotypes, the Brazilian-1 and Peruvian, have accumulated differences in pre- and post-zygotic mechanisms resulting in a degree of reproductive isolation. Both harbor a different strain of Wolbachia, which is a widespread endosymbiotic bacterium among many invertebrates producing a range of reproductive effects. In this paper, we studied the role of this bacterium as one of the factors involved in such isolation process. Infected and cured laboratory colonies were used to test pre- and post-zygotic effects, with special emphasis in uni- and bi-directional cytoplasmic incompatibility (CI). We showed that Wolbachia is the only known reproductive symbiont present in these morphotypes. Wolbachia reduced the ability for embryonic development in crosses involving cured females and infected males within each morphotype (uni-directional CI). This inhibition showed to be more effective in the Peruvian morphotype. Bi-directional CI was not evidenced, suggesting the presence of compatible Wolbachia strains. We conclude that Wolbachia is not directly involved in the speciation process of these morphotypes. Other mechanisms rather than CI should be explored in order to explain the reduced mating compatibility between the Brazilian-1 and Peruvian morphotypes.}, } @article {pmid31796568, year = {2019}, author = {Chung, M and Teigen, LE and Libro, S and Bromley, RE and Olley, D and Kumar, N and Sadzewicz, L and Tallon, LJ and Mahurkar, A and Foster, JM and Michalski, ML and Dunning Hotopp, JC}, title = {Drug Repurposing of Bromodomain Inhibitors as Potential Novel Therapeutic Leads for Lymphatic Filariasis Guided by Multispecies Transcriptomics.}, journal = {mSystems}, volume = {4}, number = {6}, pages = {}, pmid = {31796568}, issn = {2379-5077}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {To better understand the transcriptomic interplay of organisms associated with lymphatic filariasis, we conducted multispecies transcriptome sequencing (RNA-Seq) on the filarial nematode Brugia malayi, its Wolbachia endosymbiont wBm, and its laboratory vector Aedes aegypti across the entire B. malayi life cycle. In wBm, transcription of the noncoding 6S RNA suggests that it may be a regulator of bacterial cell growth, as its transcript levels correlate with bacterial replication rates. For A. aegypti, the transcriptional response reflects the stress that B. malayi infection exerts on the mosquito with indicators of increased energy demand. In B. malayi, expression modules associated with adult female samples consistently contained an overrepresentation of genes involved in chromatin remodeling, such as the bromodomain-containing proteins. All bromodomain-containing proteins encoded by B. malayi were observed to be upregulated in the adult female, embryo, and microfilaria life stages, including 2 members of the bromodomain and extraterminal (BET) protein family. The BET inhibitor JQ1(+), originally developed as a cancer therapeutic, caused lethality of adult worms in vitro, suggesting that it may be a potential therapeutic that can be repurposed for treating lymphatic filariasis.IMPORTANCE The current treatment regimen for lymphatic filariasis is mostly microfilaricidal. In an effort to identify new drug candidates for lymphatic filariasis, we conducted a three-way transcriptomics/systems biology study of one of the causative agents of lymphatic filariasis, Brugia malayi, its Wolbachia endosymbiont wBm, and its vector host Aedes aegypti at 16 distinct B. malayi life stages. B. malayi upregulates the expression of bromodomain-containing proteins in the adult female, embryo, and microfilaria stages. In vitro, we find that the existing cancer therapeutic JQ1(+), which is a bromodomain and extraterminal protein inhibitor, has adulticidal activity in B. malayi.}, } @article {pmid31793429, year = {2019}, author = {Fontes-Sousa, AP and Silvestre-Ferreira, AC and Carretón, E and Esteves-Guimarães, J and Maia-Rocha, C and Oliveira, P and Lobo, L and Morchón, R and Araújo, F and Simón, F and Montoya-Alonso, JA}, title = {Exposure of humans to the zoonotic nematode Dirofilaria immitis in Northern Portugal.}, journal = {Epidemiology and infection}, volume = {147}, number = {}, pages = {e282}, pmid = {31793429}, issn = {1469-4409}, mesh = {Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antibodies, Bacterial ; Antibodies, Helminth/*blood ; Child ; Child, Preschool ; Dirofilaria immitis/*immunology ; Dirofilariasis/*epidemiology ; Female ; Humans ; Male ; Middle Aged ; Portugal/epidemiology ; Seroepidemiologic Studies ; Wolbachia/immunology ; Young Adult ; Zoonoses/*epidemiology ; }, abstract = {Dirofilariosis caused by Dirofilaria immitis (heartworm) is a zoonosis, considered an endemic disease of dogs and cats in several countries of Western Europe, including Portugal. This study assesses the levels of D. immitis exposure in humans from Northern Portugal, to which end, 668 inhabitants of several districts belonging to two different climate areas (Csa: Bragança, Vila Real and Csb: Aveiro, Braga, Porto, Viseu) were tested for anti-D. immitis and anti-Wolbachia surface proteins (WSP) antibodies. The overall prevalence of seropositivity to both anti-D. immitis and WSP antibodies was 6.1%, which demonstrated the risk of infection with D. immitis in humans living in Northern Portugal. This study, carried out in a Western European country, contributes to the characterisation of the risk of infection with D. immitis among human population in this region of the continent. From a One Health point of view, the results of the current work also support the close relationship between dogs and people as a risk factor for human infection.}, } @article {pmid31792331, year = {2019}, author = {Zhou, JC and Li, YY and Liu, QQ and Ning, SF and Che, WN and Cong, B and Dong, H}, title = {Effects of temperature and superparasitism on quality and characteristics of thelytokous Wolbachia-infected Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) during mass rearing.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {18114}, pmid = {31792331}, issn = {2045-2322}, mesh = {Animals ; Female ; Fertility ; Genetic Fitness ; Host-Parasite Interactions/*physiology ; Hymenoptera/*growth & development/*microbiology ; Lepidoptera/parasitology ; Male ; Ovum/parasitology ; Parthenogenesis ; Pest Control, Biological/methods ; Temperature ; Wolbachia/*physiology ; }, abstract = {Thelytokous Wolbachia-infected Trichogramma spp. are widely used egg parasitoids against lepidopteran pests in biological control programs. Wolbachia may manipulate host wasps for superparasitism and is sensitive to temperature. To explore effects of temperature and superparasitism, we compared fitness parameters and Wolbachia-mediated phenotype of thelytokous Wolbachia-infected Trichogramma dendrolimi between those emerging from superparasitised or single-parasitised hosts at 17, 21, 25, or 29 °C. Infected mothers of T. dendrolimi showed reduced superparasitism and parasitism increased with temperature. Wolbachia titre decreased with temperature when females emerged from singly-parasitised hosts, but there was no correlation in superparasitised hosts. Females showed higher Wolbachia titres at 21, 25, or 29 °C when developing from superparasitised hosts. The daily male ratio of offspring increased with temperature, and the day-age threshold for 5%, 50%, or 95% daily male ratio decreased with temperature in both parasitism forms. Females that emerged from superparasitised hosts had a shorter life span and reduced fecundity. These results indicate that Wolbachia may affect host behaviour by increasing superparasitism to enhance its spread, but this has negative effects on thelytokous Wolbachia-infected T. dendrolimi.}, } @article {pmid31786246, year = {2020}, author = {Hotterbeekx, A and Raimon, S and Abd-Elfarag, G and Carter, JY and Sebit, W and Suliman, A and Siewe Fodjo, JN and De Witte, P and Logora, MY and Colebunders, R and Kumar-Singh, S}, title = {Onchocerca volvulus is not detected in the cerebrospinal fluid of persons with onchocerciasis-associated epilepsy.}, journal = {International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases}, volume = {91}, number = {}, pages = {119-123}, pmid = {31786246}, issn = {1878-3511}, mesh = {Adult ; Animals ; DNA, Helminth/cerebrospinal fluid ; Epilepsy/*parasitology ; Female ; Humans ; Male ; Microfilariae/isolation & purification ; Onchocerca volvulus/genetics/growth & development/*isolation & purification ; Onchocerciasis/cerebrospinal fluid/*complications/parasitology ; Real-Time Polymerase Chain Reaction ; Skin/parasitology ; Zebrafish ; }, abstract = {OBJECTIVES: Epidemiological evidence links onchocerciasis with the development of epilepsy. The aim of this study was to detect Onchocerca volvulus microfilariae or its bacterial endosymbiont, Wolbachia, in the cerebrospinal fluid (CSF) of persons with onchocerciasis-associated epilepsy (OAE).

METHODS: Thirteen persons with OAE and O. volvulus skin snip densities of >80 microfilariae were recruited in Maridi County (South Sudan) and their CSF obtained. Cytospin centrifuged preparations of CSF were examined by light microscopy for the presence of O. volvulus microfilariae. DNA was extracted from CSF to detect O. volvulus (O-150 repeat) by quantitative real-time PCR, and Wolbachia (FtsZ gene) by standard PCR. To further investigate whether CSF from onchocerciasis-infected participants could induce seizures, 3- and 7-day old zebrafish larvae were injected with the CSF intracardially and intraperitoneally, respectively. For other zebrafish larvae, CSF was added directly to the larval medium.

RESULTS: No microfilariae, parasite DNA, or Wolbachia DNA were detected in any of the CSF samples by light microscopy or PCR. All zebrafish survived the procedures and none developed seizures.

CONCLUSIONS: The absence of O. volvulus in the CSF suggests that OAE is likely not caused by direct parasite invasion into the central nervous system, but by another phenomenon triggered by O. volvulus infection.}, } @article {pmid31785205, year = {2020}, author = {Ullah, MS and Kamimura, T and Gotoh, T}, title = {Effects of Temperature on Demographic Parameters of Bryobia praetiosa (Acari: Tetranychidae).}, journal = {Journal of economic entomology}, volume = {113}, number = {1}, pages = {211-221}, doi = {10.1093/jee/toz294}, pmid = {31785205}, issn = {1938-291X}, mesh = {Animals ; Female ; Fertility ; Oviposition ; Reproduction ; Temperature ; *Tetranychidae ; }, abstract = {The clover mite, Bryobia praetiosa Koch (Acari: Tetranychidae), is an agricultural pest, as well as a frequent invader of hospitals and homes. However, its adaptability to different temperatures is not well understood. We used age- and stage-specific life tables to investigate the effects of temperature on demographic parameters of B. praetiosa from 15 to 35°C under a long-day photoperiod (16:8 [L:D] h). The clover mite is a thelytokous species (consisting of only females) due to its infection with the symbiotic bacterium Wolbachia. The egg-to-adult development time of female B. praetiosa decreased as the temperature increased from 15 to 32.5°C. At 35°C, females laid eggs, but no eggs hatched. The lower thermal threshold (t0) and the thermal constant (K) for egg-to-adult females were 8.7°C and 274.1 degree-days, respectively. The intrinsic optimum temperature (TØ) was 22.4°C. The oviposition period decreased with increasing temperature. Fecundity was highest at 20°C and extremely low at 30°C. The net reproductive rate (R0) decreased as the temperature increased from 15 to 30°C, but no significant difference was observed between 15 and 20°C. The intrinsic rate of natural increase (r) varied from 0.0721/d at 15°C to 0.1679/d at 25°C, and then decreased to 0.1203/d at 30°C. These results should be useful in developing management strategies for B. praetiosa.}, } @article {pmid31780680, year = {2019}, author = {Vivero, RJ and Villegas-Plazas, M and Cadavid-Restrepo, GE and Herrera, CXM and Uribe, SI and Junca, H}, title = {Wild specimens of sand fly phlebotomine Lutzomyia evansi, vector of leishmaniasis, show high abundance of Methylobacterium and natural carriage of Wolbachia and Cardinium types in the midgut microbiome.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {17746}, pmid = {31780680}, issn = {2045-2322}, mesh = {Animals ; Bacteroidetes/genetics/*isolation & purification ; Female ; Gastrointestinal Microbiome ; Humans ; Insect Vectors/*microbiology ; Leishmaniasis/*transmission ; Male ; Methylobacterium/genetics/*isolation & purification ; Psychodidae/*microbiology ; RNA, Ribosomal, 16S/genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Phlebotomine sand flies are remarkable vectors of several etiologic agents (virus, bacterial, trypanosomatid Leishmania), posing a heavy health burden for human populations mainly located at developing countries. Their intestinal microbiota is involved in a wide range of biological and physiological processes, and could exclude or facilitate such transmission of pathogens. In this study, we investigated the Eubacterial microbiome from digestive tracts of Lu. evansi adults structure using 16S rRNA gene sequence amplicon high throughput sequencing (Illumina MiSeq) obtained from digestive tracts of Lu. evansi adults. The samples were collected at two locations with high incidence of the disease in humans: peri-urban and forest ecosystems from the department of Sucre, Colombia. 289,068 quality-filtered reads of V4 region of 16S rRNA gene were obtained and clustered into 1,762 operational taxonomic units (OTUs) with 97% similarity. Regarding eubacterial diversity, 14 bacterial phyla and 2 new candidate phyla were found to be consistently associated with the gut microbiome content. Proteobacteria, Firmicutes, and Bacteroidetes were the most abundant phyla in all the samples and the core microbiome was particularly dominated by Methylobacterium genus. Methylobacterium species, are known to have mutualistic relationships with some plants and are involved in shaping the microbial community in the phyllosphere. As a remarkable feature, OTUs classified as Wolbachia spp. were found abundant on peri-urban ecosystem samples, in adult male (OTUs n = 776) and unfed female (OTUs n = 324). Furthermore, our results provide evidence of OTUs classified as Cardinium endosymbiont in relative abundance, notably higher with respect to Wolbachia. The variation in insect gut microbiota may be determined by the environment as also for the type of feeding. Our findings increase the richness of the microbiota associated with Lu. evansi. In this study, OTUs of Methylobacterium found in Lu. evansi was higher in engorged females, suggesting that there are interactions between microbes from plant sources, blood nutrients and the parasites they transmit during the blood intake.}, } @article {pmid31780533, year = {2019}, author = {Servick, K}, title = {Bacteria-armed mosquitoes make dent in dengue.}, journal = {Science (New York, N.Y.)}, volume = {366}, number = {6469}, pages = {1056}, doi = {10.1126/science.366.6469.1056}, pmid = {31780533}, issn = {1095-9203}, mesh = {Aedes/*microbiology ; Animals ; Dengue/*prevention & control ; *Dengue Virus ; *Wolbachia ; }, } @article {pmid31775631, year = {2019}, author = {Stouthamer, CM and Kelly, SE and Mann, E and Schmitz-Esser, S and Hunter, MS}, title = {Development of a multi-locus sequence typing system helps reveal the evolution of Cardinium hertigii, a reproductive manipulator symbiont of insects.}, journal = {BMC microbiology}, volume = {19}, number = {1}, pages = {266}, pmid = {31775631}, issn = {1471-2180}, mesh = {Animals ; Bacterial Typing Techniques/methods ; Bacteroidetes/*classification ; DNA, Ribosomal/genetics ; *Evolution, Molecular ; Insecta/*microbiology/physiology ; Multilocus Sequence Typing/*methods ; Phenotype ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Reproduction ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {BACKGROUND: Cardinium is an intracellular bacterial symbiont in the phylum Bacteroidetes that is found in many different species of arthropods and some nematodes. This symbiont is known to be able to induce three reproductive manipulation phenotypes, including cytoplasmic incompatibility. Placing individual strains of Cardinium within a larger evolutionary context has been challenging because only two, relatively slowly evolving genes, 16S rRNA gene and Gyrase B, have been used to generate phylogenetic trees, and consequently, the relationship of different strains has been elucidated in only its roughest form.

RESULTS: We developed a Multi Locus Sequence Typing (MLST) system that provides researchers with three new genes in addition to Gyrase B for inferring phylogenies and delineating Cardinium strains. From our Cardinium phylogeny, we confirmed the presence of a new group D, a Cardinium clade that resides in the arachnid order harvestmen (Opiliones). Many Cardinium clades appear to display a high degree of host affinity, while some show evidence of host shifts to phylogenetically distant hosts, likely associated with ecological opportunity. Like the unrelated reproductive manipulator Wolbachia, the Cardinium phylogeny also shows no clear phylogenetic signal associated with particular reproductive manipulations.

CONCLUSIONS: The Cardinium phylogeny shows evidence of diversification within particular host lineages, and also of host shifts among trophic levels within parasitoid-host communities. Like Wolbachia, the relatedness of Cardinium strains does not necessarily predict their reproductive phenotypes. Lastly, the genetic tools proposed in this study may help future authors to characterize new strains and add to our understanding of Cardinium evolution.}, } @article {pmid31774393, year = {2019}, author = {Beckmann, JF and Sharma, GD and Mendez, L and Chen, H and Hochstrasser, M}, title = {The Wolbachia cytoplasmic incompatibility enzyme CidB targets nuclear import and protamine-histone exchange factors.}, journal = {eLife}, volume = {8}, number = {}, pages = {}, pmid = {31774393}, issn = {2050-084X}, support = {R01 GM053756/GM/NIGMS NIH HHS/United States ; GM046904/GF/NIH HHS/United States ; 1015922//USDA/International ; R01 GM046904/GM/NIGMS NIH HHS/United States ; SEED Grant//Alabama Agricultural Experiment Station/International ; GM053756/GF/NIH HHS/United States ; }, mesh = {Active Transport, Cell Nucleus ; Animals ; Animals, Genetically Modified ; Centromere Protein A/genetics/*metabolism ; Crosses, Genetic ; Cytoplasm/*metabolism ; Drosophila Proteins/genetics/*metabolism ; Drosophila melanogaster/genetics/metabolism/microbiology ; Escherichia coli/metabolism ; Female ; Gene Expression Regulation ; Histones/*metabolism ; Karyopherins/genetics/metabolism ; Male ; Protamines/*metabolism ; Recombinant Proteins ; Reproduction ; Saccharomyces cerevisiae/metabolism ; Saccharomyces cerevisiae Proteins/genetics/metabolism ; Wolbachia/*metabolism ; alpha Karyopherins/isolation & purification/metabolism ; }, abstract = {Intracellular Wolbachia bacteria manipulate arthropod reproduction to promote their own inheritance. The most prevalent mechanism, cytoplasmic incompatibility (CI), traces to a Wolbachia deubiquitylase, CidB, and CidA. CidB has properties of a toxin, while CidA binds CidB and rescues embryonic viability. CidB is also toxic to yeast where we identified both host effects and high-copy suppressors of toxicity. The strongest suppressor was karyopherin-α, a nuclear-import receptor; this required nuclear localization-signal binding. A protein-interaction screen of Drosophila extracts using a substrate-trapping catalytic mutant, CidB*, also identified karyopherin-α; the P32 protamine-histone exchange factor bound as well. When CidB* bound CidA, these host protein interactions disappeared. These associations would place CidB at the zygotic male pronucleus where CI defects first manifest. Overexpression of karyopherin-α, P32, or CidA in female flies suppressed CI. We propose that CidB targets nuclear-protein import and protamine-histone exchange and that CidA rescues embryos by restricting CidB access to its targets.}, } @article {pmid31769394, year = {2020}, author = {Lau, MJ and Endersby-Harshman, NM and Axford, JK and Ritchie, SA and Hoffmann, AA and Ross, PA}, title = {Measuring the Host-Seeking Ability of Aedes aegypti Destined for Field Release.}, journal = {The American journal of tropical medicine and hygiene}, volume = {102}, number = {1}, pages = {223-231}, pmid = {31769394}, issn = {1476-1645}, support = {108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*physiology ; Animals ; Feeding Behavior/*physiology ; Female ; Mosquito Control/methods ; Pest Control, Biological/methods ; Wolbachia ; }, abstract = {Host seeking is an essential process in mosquito reproduction. Field releases of modified mosquitoes for population replacement rely on successful host seeking by female mosquitoes, but host-seeking ability is rarely tested in a realistic context. We tested the host-seeking ability of female Aedes aegypti mosquitoes using a semi-field system. Females with different Wolbachia infection types (wMel-, wAlbB-infected, and uninfected) or from different origins (laboratory and field) were released at one end of a semi-field cage and recaptured as they landed on human experimenters 15 m away. Mosquitoes from each population were then identified with molecular tools or through minimal dusting with fluorescent powder. Wolbachia-infected and uninfected populations had similar average durations to landing and overall recapture proportions, as did laboratory and field-sourced Ae. aegypti. These results indicate that the host-seeking ability of mosquitoes is not negatively affected by Wolbachia infection or long-term laboratory maintenance. This method provides an approach to study the host-seeking ability of mosquitoes in a realistic setting, which will be useful when evaluating strains of mosquitoes that are planned for releases into the field to suppress arbovirus transmission.}, } @article {pmid31767943, year = {2020}, author = {Ju, JF and Bing, XL and Zhao, DS and Guo, Y and Xi, Z and Hoffmann, AA and Zhang, KJ and Huang, HJ and Gong, JT and Zhang, X and Hong, XY}, title = {Wolbachia supplement biotin and riboflavin to enhance reproduction in planthoppers.}, journal = {The ISME journal}, volume = {14}, number = {3}, pages = {676-687}, pmid = {31767943}, issn = {1751-7370}, mesh = {Animals ; Biotin/*metabolism ; Fertility ; Genomics ; Hemiptera/*microbiology/*physiology ; Reproduction ; Riboflavin/*metabolism ; Symbiosis ; Vitamin B Complex/metabolism ; Wolbachia/genetics/*metabolism ; }, abstract = {Symbiont-mediated nutritional mutualisms can contribute to the host fitness of insects, especially for those that feed exclusively on nutritionally unbalanced diets. Here, we elucidate the importance of B group vitamins in the association of endosymbiotic bacteria Wolbachia with two plant-sap feeding insects, the small brown planthopper, Laodelphax striatellus (Fallén), and the brown planthopper, Nilaparvata lugens (Stål). Infected planthoppers of both species laid more eggs than uninfected planthoppers, while the experimental transfer of Wolbachia into uninfected lines of one planthopper species rescued this fecundity deficit. The genomic analysis showed that Wolbachia strains from the two planthopper species encoded complete biosynthesis operons for biotin and riboflavin, while a metabolic analysis revealed that Wolbachia-infected planthoppers of both species had higher titers of biotin and riboflavin. Furthermore, experimental supplementation of food with a mixture of biotin and riboflavin recovered the fecundity deficit of Wolbachia-uninfected planthoppers. In addition, comparative genomic analysis suggested that the riboflavin synthesis genes are conserved among Wolbachia supergroups. Biotin operons are rare in Wolbachia, and those described share a recent ancestor that may have been horizontally transferred from Cardinium bacteria. Our research demonstrates a type of mutualism that involves a facultative interaction between Wolbachia and plant-sap feeding insects involving vitamin Bs.}, } @article {pmid31767914, year = {2019}, author = {Liu, QQ and Zhou, JC and Zhang, C and Ning, SF and Duan, LJ and Dong, H}, title = {Co-occurrence of thelytokous and bisexual Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) in a natural population.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {17480}, pmid = {31767914}, issn = {2045-2322}, mesh = {Animals ; Behavior, Animal ; Bisexuality ; DNA, Bacterial/genetics ; DNA, Intergenic/genetics ; Electron Transport Complex IV/genetics ; Female ; Genetic Introgression ; Insect Proteins/genetics ; Male ; Phylogeny ; Wasps/*classification/microbiology/*physiology ; Wolbachia/*genetics ; }, abstract = {Trichogramma dendrolimi is one of the most successful biocontrol agents in China. However, an inundative condition is necessary to obtain acceptable parasitism effect. A good solution to this is the application of its thelytokous counterparts which unfortunately are scarce in field. We here report the first case of a natural T. dendrolimi population in China comprising both bisexual wasps and an extremely low proportion of thelytokous wasps. These two forms of T. dendrolimi are phylogenetically related based on the reconstructions of ITS-2 and COI genes. Also, the phylogenetic results suggested a potentially Wolbachia-drived ITS-2 variation. The expression of thelytoky was hardly affected by temperature, which might help control Asian corn borer and Dendrolimus punctatus. Wolbachia are responsible for current thelytoky according to phylogenetic analyses, antibiotic treatment and introgression experiment. We also present the third case of paternal sex ratio chromosome that restrains the expansion of Wolbachia. Moreover, the low frequency of thelytoky may be common in natural populations. Consequently if for biological control it is determined that a thelytokous strain is to be preferred, then large number of field collected females should be set up as isofemale lines, to detect the rare thelytoky.}, } @article {pmid31763192, year = {2019}, author = {Lucchetti, C and Genchi, M and Venco, L and Bazzocchi, C and Kramer, LH and Vismarra, A}, title = {Optimized protocol for DNA/RNA co-extraction from adults of Dirofilaria immitis.}, journal = {MethodsX}, volume = {6}, number = {}, pages = {2601-2605}, pmid = {31763192}, issn = {2215-0161}, abstract = {Dirofilaria immitis, the etiologic agent of canine heartworm disease, like several other filarial nematodes, harbors the bacterial endosymbiont Wolbachia. To investigate metabolic and functional pathways of D. immitis and Wolbachia individually, along with their interactions, the use of both transcriptomic and genome analysis has becoming increasingly popular. Although several commercial kits are available for the single extraction of either DNA or RNA, no specific protocol has been described for simultaneous extraction of DNA and RNA from such a large organism like an adult D. immitis, where female worms generally reach ∼25 cm in length. More importantly, adult worms of D. immitis can only be obtained either through necropsy of experimentally infected dogs or by minimally-invasive surgical heartworm removal of naturally infected dogs. This makes each individual worm sample extremely important. Thus, in the context of a project aimed at the evaluation of both gene expression analysis and Wolbachia population assessment following different treatments, an optimized protocol for co-extraction of DNA and RNA from a single sample of adult D. immitis has been developed. •An optimized method for DNA/RNA co-extraction from large size nematodes using TRIzol® reagent.•Allows maximum exploitation of unique samples as adults of D. immitis.}, } @article {pmid31761702, year = {2019}, author = {Nazni, WA and Hoffmann, AA and NoorAfizah, A and Cheong, YL and Mancini, MV and Golding, N and Kamarul, GMR and Arif, MAK and Thohir, H and NurSyamimi, H and ZatilAqmar, MZ and NurRuqqayah, M and NorSyazwani, A and Faiz, A and Irfan, FMN and Rubaaini, S and Nuradila, N and Nizam, NMN and Irwan, SM and Endersby-Harshman, NM and White, VL and Ant, TH and Herd, CS and Hasnor, AH and AbuBakar, R and Hapsah, DM and Khadijah, K and Kamilan, D and Lee, SC and Paid, YM and Fadzilah, K and Topek, O and Gill, BS and Lee, HL and Sinkins, SP}, title = {Establishment of Wolbachia Strain wAlbB in Malaysian Populations of Aedes aegypti for Dengue Control.}, journal = {Current biology : CB}, volume = {29}, number = {24}, pages = {4241-4248.e5}, pmid = {31761702}, issn = {1879-0445}, support = {202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 202888/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/genetics/metabolism/*microbiology ; Animals ; Dengue/*prevention & control ; Dengue Virus/metabolism/pathogenicity ; Female ; Humans ; Insect Vectors ; Malaysia ; Male ; Mosquito Vectors ; Pest Control, Biological/*methods ; Wolbachia/genetics/*metabolism ; }, abstract = {Dengue has enormous health impacts globally. A novel approach to decrease dengue incidence involves the introduction of Wolbachia endosymbionts that block dengue virus transmission into populations of the primary vector mosquito, Aedes aegypti. The wMel Wolbachia strain has previously been trialed in open releases of Ae. aegypti; however, the wAlbB strain has been shown to maintain higher density than wMel at high larval rearing temperatures. Releases of Ae. aegypti mosquitoes carrying wAlbB were carried out in 6 diverse sites in greater Kuala Lumpur, Malaysia, with high endemic dengue transmission. The strain was successfully established and maintained at very high population frequency at some sites or persisted with additional releases following fluctuations at other sites. Based on passive case monitoring, reduced human dengue incidence was observed in the release sites when compared to control sites. The wAlbB strain of Wolbachia provides a promising option as a tool for dengue control, particularly in very hot climates.}, } @article {pmid31750894, year = {2019}, author = {López-Madrigal, S and Duarte, EH}, title = {Titer regulation in arthropod-Wolbachia symbioses.}, journal = {FEMS microbiology letters}, volume = {366}, number = {23}, pages = {}, doi = {10.1093/femsle/fnz232}, pmid = {31750894}, issn = {1574-6968}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Load ; Environment ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {Symbiosis between intracellular bacteria (endosymbionts) and animals are widespread. The alphaproteobacterium Wolbachia pipientis is known to maintain a variety of symbiotic associations, ranging from mutualism to parasitism, with a wide range of invertebrates. Wolbachia infection might deeply affect host fitness (e.g. reproductive manipulation and antiviral protection), which is thought to explain its high prevalence in nature. Bacterial loads significantly influence both the infection dynamics and the extent of bacteria-induced host phenotypes. Hence, fine regulation of bacterial titers is considered as a milestone in host-endosymbiont interplay. Here, we review both environmental and biological factors modulating Wolbachia titers in arthropods.}, } @article {pmid31747432, year = {2019}, author = {Jasper, ME and Yang, Q and Ross, PA and Endersby-Harshman, N and Bell, N and Hoffmann, AA}, title = {A LAMP assay for the rapid and robust assessment of Wolbachia infection in Aedes aegypti under field and laboratory conditions.}, journal = {PloS one}, volume = {14}, number = {11}, pages = {e0225321}, pmid = {31747432}, issn = {1932-6203}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; DNA, Bacterial/chemistry/genetics ; Nucleic Acid Amplification Techniques/*methods/standards ; Sensitivity and Specificity ; Wolbachia/*genetics/pathogenicity ; }, abstract = {With Wolbachia-based arbovirus control programs being scaled and operationalised around the world, cost effective and reliable detection of Wolbachia in field samples and laboratory stocks is essential for quality control. Here we validate a modified loop-mediated isothermal amplification (LAMP) assay for routine scoring of Wolbachia in mosquitoes from laboratory cultures and the field, applicable to any setting. We show that this assay is a rapid and robust method for highly sensitive and specific detection of wAlbB Wolbachia infection within Aedes aegypti under a variety of conditions. We test the quantitative nature of the assay by evaluating pooled mixtures of Wolbachia-infected and uninfected mosquitoes and show that it is capable of estimating infection frequencies, potentially circumventing the need to perform large-scale individual analysis for wAlbB infection status in the course of field monitoring. These results indicate that LAMP assays are useful for routine screening particularly under field conditions away from laboratory facilities.}, } @article {pmid31736900, year = {2019}, author = {Zhao, C and Zhao, H and Zhang, S and Luo, J and Zhu, X and Wang, L and Zhao, P and Hua, H and Cui, J}, title = {The Developmental Stage Symbionts of the Pea Aphid-Feeding Chrysoperla sinica (Tjeder).}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {2454}, pmid = {31736900}, issn = {1664-302X}, abstract = {Chrysoperla sinica (Tjeder) is widely recognized as an important holometabolous natural enemy of various insect pests in different cropping systems and as a non-target surrogate in environmental risk assessment of Bt rice (i.e., genetically modified rice to express a toxin gene from Bacillus thuringiensis). Like other complex organisms, abundant microbes live inside C. sinica; however, to date, microbiome composition and diversity of the whole life cycle in C. sinica has not yet been well characterized. In the current study, we analyze the composition and biodiversity of microbiota across the whole life cycle of C. sinica by using high-throughput Illumina sequencing of the 16S ribosomal RNA gene. Collectively, Proteobacteria and Firmicutes dominated the microenvironment at all stages, but their relative abundances fluctuated by host developmental stage. Interestingly, eggs, neonates, and adults shared similar microbes, including an abundance of Rickettsia and Wolbachia. After larva feeding, Staphylococcus, Enterobacteriaceae, and Serratia were enriched in larvae and pupa, suggesting that food may serve as a major factor contributing to altered microbial community divergence at different developmental stages. Our findings demonstrated that C. sinica harbor a variety of bacteria, and that dynamic changes in community composition and relative abundances of members of its microbiome occur during different life cycle stages. Evaluating the role of these bacterial symbionts in this natural enemy may assist in developing environmental risk assessments and novel biological control strategies.}, } @article {pmid31726321, year = {2020}, author = {Ote, M and Yamamoto, D}, title = {Impact of Wolbachia infection on Drosophila female germline stem cells.}, journal = {Current opinion in insect science}, volume = {37}, number = {}, pages = {8-15}, doi = {10.1016/j.cois.2019.10.001}, pmid = {31726321}, issn = {2214-5753}, mesh = {Animals ; Bacterial Proteins/metabolism ; Drosophila Proteins/metabolism ; Drosophila melanogaster/genetics/*microbiology ; Female ; Oogonial Stem Cells/metabolism/*microbiology ; RNA/metabolism ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis, one of the most dominant insect-symbiotic bacteria, highjacks the female germline of insects for its own propagation across host generations. Such strict dependence on female gametes in trans-generational propagation has driven Wolbachia to devise ingenious strategies to enhance female fertility. In Drosophila melanogaster females with female-sterile mutant alleles of the master sex-determining gene Sex-lethal (Sxl), Wolbachia colonizing female germline stem cells (GSCs) support the maintenance of GSCs, thereby rescuing the defective ovarian development. In the germ cell cytoplasm, Wolbachia are often found in proximity to ribonucleoprotein-complex processing bodies (P bodies), where the Wolbachia-derived protein TomO interacts with RNAs encoding Nanos and Orb proteins, which support the GSC maintenance and oocyte polarization, respectively. Thus, manipulation of host RNA is the key to successful vertical transmission of Wolbachia.}, } @article {pmid31723729, year = {2019}, author = {, }, title = {The World Health Organization 2030 goals for onchocerciasis: Insights and perspectives from mathematical modelling: NTD Modelling Consortium Onchocerciasis Group.}, journal = {Gates open research}, volume = {3}, number = {}, pages = {1545}, pmid = {31723729}, issn = {2572-4754}, abstract = {The World Health Organization (WHO) has embarked on a consultation process to refine the 2030 goals for priority neglected tropical diseases (NTDs), onchocerciasis among them. Current goals include elimination of transmission (EOT) by 2020 in Latin America, Yemen and selected African countries. The new goals propose that, by 2030, EOT be verified in 10 countries; mass drug administration (MDA) with ivermectin be stopped in at least one focus in 34 countries; and that the proportion of the population no longer in need of MDA be equal or greater than 25%, 50%, 75% and 100% in at least 16, 14, 12, and 10 countries, respectively. The NTD Modelling Consortium onchocerciasis teams have used EPIONCHO and ONCHOSIM to provide modelling insights into these goals. EOT appears feasible in low-moderate endemic areas with long-term MDA at high coverage (≥75%), but uncertain in areas of higher endemicity, poor coverage and adherence, and where MDA has not yet, or only recently, started. Countries will have different proportions of their endemic areas classified according to these categories, and this distribution of pre-intervention prevalence and MDA duration and programmatic success will determine the feasibility of achieving the proposed MDA cessation goals. Highly endemic areas would benefit from switching to biannual or quarterly MDA and implementing vector control where possible (determining optimal frequency and duration of anti-vectorial interventions requires more research). Areas without loiasis that have not yet initiated MDA should implement biannual (preferably with moxidectin) or quarterly MDA from the start. Areas with loiasis not previously treated would benefit from implementing test-and(not)-treat-based interventions, vector control, and anti- Wolbachia therapies, but their success will depend on the levels of screening and coverage achieved and sustained. The diagnostic performance of IgG4 Ov16 serology for assessing EOT is currently uncertain. Verification of EOT requires novel diagnostics at the individual- and population-levels.}, } @article {pmid31715700, year = {2019}, author = {Yin, X and Zhao, S and Yan, B and Tian, Y and Ba, T and Zhang, J and Wang, Y}, title = {Bartonella rochalimae, B. grahamii, B. elizabethae, and Wolbachia spp. in Fleas from Wild Rodents near the China-Kazakhstan Border.}, journal = {The Korean journal of parasitology}, volume = {57}, number = {5}, pages = {553-559}, pmid = {31715700}, issn = {1738-0006}, support = {2016IK264//General Administration of Quality Supervision, Inspection and Quarantine/ ; 81560338//National Natural Science Foundation of China/ ; 2018ZX10101002-007//National Key Research and Development Program of China/ ; }, mesh = {Animals ; Animals, Wild/microbiology ; Bartonella/classification/genetics/*isolation & purification ; Bartonella Infections/microbiology/*veterinary ; China ; Genotype ; Kazakhstan ; Phylogeny ; Rodent Diseases/*microbiology ; Rodentia/classification/*microbiology ; Siphonaptera/classification/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {The Alataw Pass, near the Ebinur Lake Wetland (northwest of China) and Taldykorgan (east of Kazakhstan), is a natural habitat for wild rodents. To date, little has been done on the surveillance of Bartonella spp. and Wolbachia spp. from fleas in the region. Here we molecularly detected Bartonella spp. and Wolbachia spp. in wild rodent fleas during January and October of 2016 along the Alataw Pass-Kazakhstan border. A total of 1,706 fleas belonging to 10 species were collected from 6 rodent species. Among the 10 flea species, 4 were found to be positive for Wolbachia, and 5 flea species were positive for Bartonella. Molecular analysis indicated that i) B. rochalimae was firstly identified in Xenopsylla gerbilli minax and X. conforms conforms, ii) B. grahamii was firstly identified in X. gerbilli minax, and iii) B. elizabethae was firstly detected in Coptopsylla lamellifer ardua, Paradoxopsyllus repandus, and Nosopsyllus laeviceps laeviceps. Additionally, 3 Wolbachia endosymbionts were firstly found in X. gerbilli minax, X. conforms conforms, P. repandus, and N. laeviceps laeviceps. BLASTn analysis indicated 3 Bartonella species showed genotypic variation. Phylogenetic analysis revealed 3 Wolbachia endosymbionts were clustered into the non-Siphonaptera Wolbachia group. These findings extend our knowledge of the geographical distribution and carriers of B. rochalimae, B. grahamii, B. elizabethae, and Wolbachia spp. In the future, there is a need for China-Kazakhstan cooperation to strengthen the surveillance of flea-borne pathogens in wildlife.}, } @article {pmid31709323, year = {2019}, author = {Adekunle, AI and Meehan, MT and McBryde, ES}, title = {Mathematical analysis of a Wolbachia invasive model with imperfect maternal transmission and loss of Wolbachia infection.}, journal = {Infectious Disease Modelling}, volume = {4}, number = {}, pages = {265-285}, pmid = {31709323}, issn = {2468-0427}, abstract = {Arboviral infections, especially dengue, continue to cause significant health burden in their endemic regions. One of the strategies to tackle these infections is to replace the main vector agent, Ae. aegypti, with the ones incapable of transmitting the virus. Wolbachia, an intracellular bacterium, has shown promise in achieving this goal. However, key factors such as imperfect maternal transmission, loss of Wolbachia infection, reduced reproductive capacity and shortened life-span affect the dynamics of Wolbachia in different forms in the Ae. aegypti population. In this study, we developed a Wolbachia transmission dynamic model adjusting for imperfect maternal transmission and loss of Wolbachia infection. The invasive reproductive number that determines the likelihood of replacement of the Wolbachia-uninfected (WU) population is derived and with it, we established the local and global stability of the equilibrium points. This analysis clearly shows that cytoplasmic incompatibility (CI) does not guarantee establishment of the Wolbachia-infected (WI) mosquitoes as imperfect maternal transmission and loss of Wolbachia infection could outweigh the gains from CI. Optimal release programs depending on the level of imperfect maternal transmission and loss of Wolbachia infection are shown. Hence, it is left to decision makers to either aim for replacement or co-existence of both populations.}, } @article {pmid31699757, year = {2019}, author = {Basting, PJ and Bergman, CM}, title = {Complete Genome Assemblies for Three Variants of the Wolbachia Endosymbiont of Drosophila melanogaster.}, journal = {Microbiology resource announcements}, volume = {8}, number = {45}, pages = {}, pmid = {31699757}, issn = {2576-098X}, abstract = {Here, we report genome assemblies for three strains of Wolbachia pipientis, assembled from unenriched, unfiltered long-read shotgun sequencing data of geographically distinct strains of Drosophila melanogaster Our simple methodology can be applied to long-read data sets of other Wolbachia-infected species with limited Wolbachia-host lateral gene transfers to produce complete assemblies for this important model symbiont.}, } @article {pmid31698562, year = {2019}, author = {Almeida, L and Duprez, MG and Privat, Y and Vauchelet, N}, title = {Mosquito population control strategies for fighting against arboviruses.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {16}, number = {6}, pages = {6274-6297}, doi = {10.3934/mbe.2019313}, pmid = {31698562}, issn = {1551-0018}, mesh = {Aedes ; Algorithms ; Animals ; Arbovirus Infections/*prevention & control/therapy ; Arboviruses ; Computer Simulation ; Culicidae/microbiology/virology ; Female ; Life Cycle Stages ; Male ; Models, Statistical ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/*virology ; Pest Control, Biological/*methods ; Population Dynamics ; Probability ; Wolbachia ; }, abstract = {In the fight against vector-borne arboviruses, an important strategy of control of epidemic consists in controlling the population of the vector, Aedes mosquitoes in this case. Among possible actions, two techniques consist either in releasing sterile mosquitoes to reduce the size of the population (Sterile Insect Technique) or in replacing the wild population by one carrying a bacteria, called Wolbachia, blocking the transmission of viruses from insects to humans. This article addresses the issue of optimizing the dissemination protocol for each of these strategies, in order to get as close as possible to these objectives. Starting from a mathematical model describing population dynamics, we study the control problem and introduce the cost function standing for population replacement and sterile insect technique. Then, we establish some properties of the optimal control and illustrate them with numerical simulations.}, } @article {pmid31698114, year = {2020}, author = {Karut, K and Castle, SJ and Karut, ŞT and Karaca, MM}, title = {Secondary endosymbiont diversity of Bemisia tabaci and its parasitoids.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {78}, number = {}, pages = {104104}, doi = {10.1016/j.meegid.2019.104104}, pmid = {31698114}, issn = {1567-7257}, mesh = {Animals ; Arizona ; Bacteria/*classification/genetics/isolation & purification ; Gammaproteobacteria/genetics/isolation & purification ; Genotyping Techniques/*methods ; Hemiptera/*classification/genetics/*microbiology ; Mediterranean Region ; Phylogeography ; Rickettsia/genetics/isolation & purification ; Species Specificity ; Symbiosis ; Turkey ; Wolbachia/genetics/isolation & purification ; }, abstract = {Cotton whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is one of the most important insect pests worldwide. It is known as a species complex consisting of at least 40 cryptic species. Although there are substantial data regarding species composition, parasitoids and endosymbionts of B. tabaci, data on relationship between the pest, parasitoids and endosymbionts are very restricted. Therefore, in this study, secondary endosymbionts in populations of B. tabaci and their parasitoids collected from Turkey and the USA were determined by PCR-based DNA analysis. Whitefly populations in Turkey represented both Mediterranean (MED) and Middle East-Asia Minor1 (MEAM1) genotypes from single or mixed populations of both genotypes. Arsenophonus, Rickettsia and Wolbachia were found in MED, while Hamiltonella and Rickettsia in MEAM1. Whitefly populations collected from Arizona were all MEAM1 and dually infected with Hamiltonella and Rickettsia. The aphelinid parasitoids Encarsia lutea and Eretmocerus mundus predominated in all Turkish populations. While almost all En. lutea populations were infected with Wolbachia, no endosymbionts were detected in any Er. mundus. Parasitoid species and the pattern of secondary endosymbiont infection in Arizona populations were different with Rickettsia detected only from Encarsia sophia while both Rickettsia and Wolbachia were found in Eretmocerus species. As a result, four secondary endosymbionts, namely, Rickettsia, Hamiltonella, Arsenophonus and Wolbachia, were detected from B.tabaci and its parasitoids. Among them only Wolbachia and Rickettsia were found in both the pest and parasitoids. It is conclude that further studies should be pursued to determine effect of these endosymbionts on biology of the parasitoids and success in biological control of B. tabaci.}, } @article {pmid31690673, year = {2019}, author = {Layton, EM and On, J and Perlmutter, JI and Bordenstein, SR and Shropshire, JD}, title = {Paternal Grandmother Age Affects the Strength of Wolbachia-Induced Cytoplasmic Incompatibility in Drosophila melanogaster.}, journal = {mBio}, volume = {10}, number = {6}, pages = {}, pmid = {31690673}, issn = {2150-7511}, support = {R21 HD086833/HD/NICHD NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; R21 AI133522/AI/NIAID NIH HHS/United States ; F31 AI143152/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/*genetics/*microbiology ; Drosophila melanogaster/*genetics/*microbiology ; Female ; Grandparents ; Male ; Reproduction/genetics ; Wolbachia/*genetics ; }, abstract = {Wolbachia are obligate intracellular bacteria that are globally distributed in half of all arthropod species. As the most abundant maternally inherited microbe in animals, Wolbachia manipulate host reproduction via reproductive parasitism strategies, including cytoplasmic incompatibility (CI). CI manifests as embryonic death when Wolbachia-modified sperm fertilize uninfected eggs but not maternally infected eggs. Thus, CI can provide a relative fitness advantage to Wolbachia-infected females and drive the infection through a population. In the genetic model Drosophila melanogaster, the Wolbachia strain wMel induces variable CI, making mechanistic studies in D. melanogaster cumbersome. Here, we demonstrate that sons of older paternal D. melanogaster grandmothers induce stronger CI than sons of younger paternal grandmothers, and we term this relationship the "paternal grandmother age effect" (PGAE). Moreover, the embryos and adult sons of older D. melanogaster grandmothers have higher Wolbachia densities, correlating with their ability to induce stronger CI. In addition, we report that Wolbachia density positively correlates with female age and decreases after mating, suggesting that females transmit Wolbachia loads that are proportional to their own titers. These findings reveal a transgenerational impact of age on wMel-induced CI, elucidate Wolbachia density dynamics in D. melanogaster, and provide a methodological advance to studies aimed at understanding wMel-induced CI in the D. melanogaster model.IMPORTANCE Unidirectional cytoplasmic incompatibility (CI) results in a postfertilization incompatibility between Wolbachia-infected males and uninfected females. CI contributes to reproductive isolation between closely related species and is used in worldwide vector control programs to drastically lower arboviral vector population sizes or to replace populations that transmit arboviruses with those resistant to transmission. Despite decades of research on the factors that influence CI, penetrance is often variable under controlled laboratory conditions in various arthropods, suggesting that additional variables influence CI strength. Here, we demonstrate that paternal D. melanogaster grandmother age influences the strength of CI induced by their sons. Older D. melanogaster females have higher Wolbachia densities and produce offspring with higher Wolbachia densities that associate with stronger CI. This work reveals a multigenerational impact of age on CI and expands our understanding of host-Wolbachia interactions and the biology of CI induced by the Wolbachia strain infecting the most widely used arthropod model, D. melanogaster.}, } @article {pmid31683719, year = {2019}, author = {Liang, Y and Hong, Y and Mai, Z and Zhu, Q and Guo, L}, title = {Internal and External Microbial Community of the Thitarodes Moth, the Host of Ophiocordyceps sinensis.}, journal = {Microorganisms}, volume = {7}, number = {11}, pages = {}, pmid = {31683719}, issn = {2076-2607}, support = {No. 2018A030313094//Natural Science Foundation of Guangdong Province/ ; No. 20185071521641//Social Science and Technology Development Project of Dongguan/ ; No. 81303155//National Natural Science Foundation of China/ ; No. 2018GkQNCX050//Project for Young Innovative Talents in Ordinary Higher University of Guangdong Province/ ; }, abstract = {Ophiocordyceps sinensis is a widely known medicinal entomogenous fungus, which parasitizes the soil-borne larva of Thitarodes (Hepialidae, Lepidoptera) distributed in the Qinghai-Tibetan Plateau and its adjacent areas. Previous research has involved artificial cultivation of Chinese cordyceps (the fungus-caterpillar complex), but it is difficult to achieve large-scale cultivation because the coupling relation between the crucial microbes and their hosts is not quite clear. To clarify the influence of the internal microbial community on the occurrence of Chinese cordyceps, in this study, the unfertilized eggs of Thitarodes of different sampling sites were chosen to analyze the bacterial and fungal communities via 16S rRNA and ITS sequencing for the first time. The results showed that for bacteria, 348 genera (dominant genera include Wolbachia, Spiroplasma, Carnobacterium, Sphingobium, and Acinetobacter) belonging to 26 phyla (dominant phyla include Proteobacteria, Firmicutes, Tenericutes, Actinobacteria, Acidobacteria, and Bacteroidetes), 58 classes, 84 orders, and 120 families were identified from 1294 operational taxonomic units (OTUs). The dominant bacterial genus (Spiroplasma) may be an important bacterial factor promoting the occurrence of Chinese cordyceps. For fungi, 289 genera, mainly including Aureobasidium, Candida, and Cryptococcus, were identified, and they belonged to 5 phyla (Ascomycota, Basidiomycota, Chytridiomycota, Glomeromycota, and Zygomycota), 26 classes, 82 orders, and 165 families. Eight bacterial OTUs and 12 fungal OTUs were shared among all of the detected samples and were considered as core species. Among them, Wolbachia, Spiroplasma, Carnobacterium, Aureobasidium, and Phoma may play important roles in helping the host larva to digest foods, adapt to extreme environments, or resist pathogens. On the other hand, the external (soil) microbial community was synchronously and comparatively analyzed. Comparative analysis revealed that external microbial factors might play a more significant role in the occurrence of Chinese cordyceps, owing to the significant differences revealed by α-diversity and β-diversity analyses among different groups. In summary, the results of this study may contribute to the large-scale cultivation of Chinese cordyceps.}, } @article {pmid31665056, year = {2019}, author = {Kieran, TJ and Arnold, KMH and Thomas, JC and Varian, CP and Saldaña, A and Calzada, JE and Glenn, TC and Gottdenker, NL}, title = {Regional biogeography of microbiota composition in the Chagas disease vector Rhodnius pallescens.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {504}, pmid = {31665056}, issn = {1756-3305}, support = {DGE-1545433//National Science Foundation/ ; COL11-043//Secretaría Nacional de Ciencia, Tecnología e Innovación/ ; }, mesh = {Actinobacteria/classification/genetics ; Analysis of Variance ; Animals ; Bacteria/*classification/genetics ; Bacteroidetes/classification/genetics ; Biodiversity ; Chagas Disease/*transmission ; DNA Barcoding, Taxonomic ; Ecosystem ; Firmicutes/classification/genetics ; Gene Library ; Humans ; Insect Vectors/*microbiology/physiology ; *Microbiota ; Panama ; Phylogeography ; Polymerase Chain Reaction ; Proteobacteria/classification/genetics ; RNA, Ribosomal, 16S/chemistry ; Rhodnius/*microbiology/physiology ; }, abstract = {BACKGROUND: Triatomine bugs are vectors of the protozoan parasite Trypanosoma cruzi, which causes Chagas disease. Rhodnius pallescens is a major vector of Chagas disease in Panama. Understanding the microbial ecology of disease vectors is important in the development of vector management strategies that target vector survival and fitness. In this study we examined the whole-body microbial composition of R. pallescens from three locations in Panama.

METHODS: We collected 89 R. pallescens specimens using Noireau traps in Attalea butyracea palms. We then extracted total DNA from whole-bodies of specimens and amplified bacterial microbiota using 16S rRNA metabarcoding PCR. The 16S libraries were sequenced on an Illumina MiSeq and analyzed using QIIME2 software.

RESULTS: We found Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes to be the most abundant bacterial phyla across all samples. Geographical location showed the largest difference in microbial composition with northern Veraguas Province having the most diversity and Panama Oeste Province localities being most similar to each other. Wolbachia was detected in high abundance (48-72%) at Panama Oeste area localities with a complete absence of detection in Veraguas Province. No significant differences in microbial composition were detected between triatomine age class, primary blood meal source, or T. cruzi infection status.

CONCLUSIONS: We found biogeographical regions differ in microbial composition among R. pallescens populations in Panama. While overall the microbiota has bacterial taxa consistent with previous studies in triatomine microbial ecology, locality differences are an important observation for future studies. Geographical heterogeneity in microbiomes of vectors is an important consideration for future developments that leverage microbiomes for disease control.}, } @article {pmid31662085, year = {2019}, author = {Martinez, J and Bruner-Montero, G and Arunkumar, R and Smith, SCL and Day, JP and Longdon, B and Jiggins, FM}, title = {Virus evolution in Wolbachia-infected Drosophila.}, journal = {Proceedings. Biological sciences}, volume = {286}, number = {1914}, pages = {20192117}, pmid = {31662085}, issn = {1471-2954}, support = {/WT_/Wellcome Trust/United Kingdom ; 094664/WT_/Wellcome Trust/United Kingdom ; 281668/ERC_/European Research Council/International ; }, mesh = {Aedes ; Animals ; Culicidae ; Drosophila/*microbiology/physiology ; *Symbiosis ; Virus Diseases ; *Viruses ; Wolbachia/*physiology ; }, abstract = {Wolbachia, a common vertically transmitted symbiont, can protect insects against viral infection and prevent mosquitoes from transmitting viral pathogens. For this reason, Wolbachia-infected mosquitoes are being released to prevent the transmission of dengue and other arboviruses. An important question for the long-term success of these programmes is whether viruses can evolve to escape the antiviral effects of Wolbachia. We have found that Wolbachia altered the outcome of competition between strains of the DCV virus in Drosophila. However, Wolbachia still effectively blocked the virus genotypes that were favoured in the presence of the symbiont. We conclude that Wolbachia did cause an evolutionary response in viruses, but this has little or no impact on the effectiveness of virus blocking.}, } @article {pmid31659008, year = {2020}, author = {Newton, ILG and Rice, DW}, title = {The Jekyll and Hyde Symbiont: Could Wolbachia Be a Nutritional Mutualist?.}, journal = {Journal of bacteriology}, volume = {202}, number = {4}, pages = {}, pmid = {31659008}, issn = {1098-5530}, mesh = {Adenosine Triphosphate/metabolism ; Biological Transport ; Genome, Bacterial ; Heme/metabolism ; Nutritional Physiological Phenomena ; Symbiosis/*physiology ; Wolbachia/genetics/*physiology ; }, abstract = {The most common intracellular symbiont on the planet-Wolbachia pipientis-is infamous largely for the reproductive manipulations induced in its host. However, more recent evidence suggests that this bacterium may also serve as a nutritional mutualist in certain host backgrounds and for certain metabolites. We performed a large-scale analysis of conserved gene content across all sequenced Wolbachia genomes to infer potential nutrients made by these symbionts. We review and critically evaluate the prior research supporting a beneficial role for Wolbachia and suggest future experiments to test hypotheses of metabolic provisioning.}, } @article {pmid31658265, year = {2019}, author = {Kittayapong, P and Ninphanomchai, S and Limohpasmanee, W and Chansang, C and Chansang, U and Mongkalangoon, P}, title = {Combined sterile insect technique and incompatible insect technique: The first proof-of-concept to suppress Aedes aegypti vector populations in semi-rural settings in Thailand.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {10}, pages = {e0007771}, pmid = {31658265}, issn = {1935-2735}, mesh = {Aedes/microbiology/*physiology/radiation effects ; Animals ; Entomology/*methods ; Female ; Humans ; Infertility, Male ; Male ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/*physiology/radiation effects ; Population Dynamics ; *Rural Population ; Sex Characteristics ; Thailand ; Wolbachia/genetics/physiology ; }, abstract = {BACKGROUND: Important arboviral diseases, such as dengue, chikungunya, and Zika virus infections, are transmitted mainly by the Aedes aegypti vector. So far, controlling this vector species with current tools and strategies has not demonstrated sustainable and significant impacts. Our main objective was to evaluate whether open field release of sterile males, produced from combining the sterile insect technique using radiation with the insect incompatible technique through Wolbachia-induced incompatibility (SIT/IIT), could suppress natural populations of Ae. aegypti in semi-rural village settings in Thailand.

Irradiated Wolbachia-infected Aedes aegypti males produced by the SIT/IIT approach were completely sterile and were able to compete with the wild fertile ones. Open field release of these sterile males was conducted in an ecologically isolated village in Chachoengsao Province, eastern Thailand. House-to-house visit and media reports resulted in community acceptance and public awareness of the technology. During intervention, approximately 100-200 sterile males were released weekly in each household. After 6 months of sterile male release, a significant reduction (p<0.05) of the mean egg hatch rate (84%) and the mean number of females per household (97.30%) was achieved in the treatment areas when compared to the control ones.

CONCLUSIONS/SIGNIFICANCE: Our study represents the first open field release of sterile Ae. aegypti males developed from a combined SIT/IIT approach. Entomological assessment using ovitraps, adult sticky traps, and portable vacuum aspirators confirmed the success in reducing natural populations of Ae. aegypti females in treated areas. Public awareness through media resulted in positive support for practical use of this strategy in wider areas. Further study using a systematic randomized trial is needed to determine whether this approach could have a significant impact on the diseases transmitted by Ae. aegypti vector.}, } @article {pmid31649084, year = {2019}, author = {Gasser, MT and Chung, M and Bromley, RE and Nadendla, S and Dunning Hotopp, JC}, title = {Complete Genome Sequence of wAna, the Wolbachia Endosymbiont of Drosophila ananassae.}, journal = {Microbiology resource announcements}, volume = {8}, number = {43}, pages = {}, pmid = {31649084}, issn = {2576-098X}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {Here, we present the complete genome sequence of the Wolbachia endosymbiont wAna, isolated from Drosophila ananassae and derived from Oxford Nanopore and Illumina sequencing. We anticipate that this will aid in Wolbachia comparative genomics and the assembly of D. ananassae specifically in regions containing extensive lateral gene transfer events.}, } @article {pmid31647706, year = {2019}, author = {Staunton, KM and Usher, L and Prachar, T and Ritchie, SA and Snoad, N and Johnson, BJ}, title = {A Novel Methodology For Recording Wing Beat Frequencies of Untethered Male and Female Aedes aegypti.}, journal = {Journal of the American Mosquito Control Association}, volume = {35}, number = {3}, pages = {169-177}, doi = {10.2987/18-6799.1}, pmid = {31647706}, issn = {1943-6270}, mesh = {Aedes/*physiology ; Animals ; Entomology/methods ; Female ; Male ; Mosquito Control/*methods ; Mosquito Vectors/*physiology ; Wings, Animal/*physiology ; }, abstract = {Aedes aegypti is a vector of many significant arboviruses worldwide, including dengue, Zika, chikungunya, and yellow fever viruses. With vector control methodology pivoting toward rearing and releasing large numbers of insects for either population suppression or virus-blocking, economical remote (sentinel) surveillance methods for release tracking become increasingly necessary. Recent steps in this direction include advances in optical sensors that identify and classify insects based on their wing beat frequency (WBF). As these traps are being developed, there is a strong need to better understand the environmental and biological factors influencing mosquito WBFs. Here, we developed new untethered-subject methodology to detect changes in WBFs of male and female Ae. aegypti. This new methodology involves directing an ultrasonic transducer at a free-flying subject and measuring the Doppler shift of the reflected ultrasonic continuous wave signal. This system's utility was assessed by determining its ability to confirm previous reports on the effect of temperature, body size, and age on the WBFs generated from acoustic or optical-based experiments. The presented ultrasonic method successfully detected expected trends for each factor for both male and female Ae. aegypti without the need for subject manipulation and potential impediment of natural flight dynamics due to tethering. As a result, this ultrasonic methodology provides a new method for understanding the environmental and physiological determinants of male and female WBFs that can inform the design of remote mosquito surveillance systems.}, } @article {pmid31644794, year = {2019}, author = {Chen, R and Su, X and Chen, J and Jiang, L and Qiao, GX}, title = {Wolbachia Infection in Two Species: Novel Views on the Colonization Ability of Wolbachia in Aphids.}, journal = {Environmental entomology}, volume = {48}, number = {6}, pages = {1388-1393}, doi = {10.1093/ee/nvz122}, pmid = {31644794}, issn = {1938-2936}, mesh = {Animals ; *Aphids ; China ; Geography ; *Infections ; *Wolbachia ; }, abstract = {Wolbachia pipientis (Rickettsiales: Anaplasmataceae) is an intracellular symbiont residing in arthropods and filarial nematodes. Sixteen supergroups have been described from different host taxa. Four supergroups A, B, M, and N were found in aphids according to prior studies. The cotton aphid, Aphis gossypii, and the green peach aphid, Myzus persicae, are typical polyphagous species with global distributions. We conducted an extensive and systematic survey of Wolbachia infections in these aphids from China. High incidences of Wolbachia infection were detected. The total infection incidence was 60% in A. gossypii and 88% in M. persicae. Both aphid species were infected with supergroups A, B and M. Different incidences of infection were observed among the seven geographical regions in China, which suggested a positive relationship between Wolbachia infections and the geographical distribution of aphid species. Furthermore, multiple infection patterns (M, B, A&M, B&M, and A&B&M) were observed. Infection patterns M and B&M were detected in almost all populations. Patterns A&B&M and B showed geographical restriction in North China. Three factors can possibly influence the Wolbachia infection incidences and patterns: the geographical distribution, aphid species, and different supergroup types.}, } @article {pmid31641465, year = {2019}, author = {Gómez-Zurita, J}, title = {Assessment of the role of Wolbachia in mtDNA paraphyly and the evolution of unisexuality in Calligrapha (Coleoptera: Chrysomelidae).}, journal = {Ecology and evolution}, volume = {9}, number = {19}, pages = {11198-11214}, pmid = {31641465}, issn = {2045-7758}, abstract = {Calligrapha is a New World leaf beetle genus that includes several unisexual species in northeastern North America. Each unisexual species had an independent hybrid origin involving different combinations of bisexual species. However, surprisingly, they all cluster in a single mtDNA clade and with some individuals of their parental species, which are in turn deeply polyphyletic for mtDNA. This pattern is suggestive of a selective sweep which, together with mtDNA taxonomic incongruence and occurrence of unisexuality in Calligrapha, led to hypothesize that Wolbachia might be responsible. I tested this hypothesis studying the correlation between diversity of Wolbachia and well-established mtDNA lineages in >500 specimens of two bisexual species of Calligrapha and their derived unisexual species. Wolbachia appears highly prevalent (83.4%), and fifteen new supergroup-A strains of the bacteria are characterized, belonging to three main classes: wCallA, occupying the whole species ranges, and wCallB and wCallC, narrowly parapatric, infecting beetles with highly divergent mtDNAs where they coexist. Most beetles (71.6%) carried double infections of wCallA with another sequence class. Bayesian inference of ancestral character states and association tests between bacterial diversity and the mtDNA genealogy show that each mtDNA lineage of Calligrapha has specific types of infection. Moreover, shifts can be explained by horizontal or vertical transfer from local populations to an expanding lineage and cytoplasmic incompatibility between wCallB and wCallC types, suggesting that the symbionts hitchhike with the host and are not responsible for selective mtDNA sweeps. Lack of evidence for sweeps and the fact that individuals in the unisexual clade are uninfected or infected by the widespread wCallA type indicate that Wolbachia does not induce unisexuality in Calligrapha, although they may manipulate host reproduction through cytoplasmic incompatibility.}, } @article {pmid31632650, year = {2019}, author = {König, K and Zundel, P and Krimmer, E and König, C and Pollmann, M and Gottlieb, Y and Steidle, JLM}, title = {Reproductive isolation due to prezygotic isolation and postzygotic cytoplasmic incompatibility in parasitoid wasps.}, journal = {Ecology and evolution}, volume = {9}, number = {18}, pages = {10694-10706}, pmid = {31632650}, issn = {2045-7758}, abstract = {The reproductive barriers that prevent gene flow between closely related species are a major topic in evolutionary research. Insect clades with parasitoid lifestyle are among the most species-rich insects and new species are constantly described, indicating that speciation occurs frequently in this group. However, there are only very few studies on speciation in parasitoids. We studied reproductive barriers in two lineages of Lariophagus distinguendus (Chalcidoidea: Hymenoptera), a parasitoid wasp of pest beetle larvae that occur in human environments. One of the two lineages occurs in households preferably attacking larvae of the drugstore beetle Stegobium paniceum ("DB-lineage"), the other in grain stores with larvae of the granary weevil Sitophilus granarius as main host ("GW-lineage"). Between two populations of the DB-lineage, we identified slight sexual isolation as intraspecific barrier. Between populations from both lineages, we found almost complete sexual isolation caused by female mate choice, and postzygotic isolation, which is partially caused by cytoplasmic incompatibility induced by so far undescribed endosymbionts which are not Wolbachia or Cardinium. Because separation between the two lineages is almost complete, they should be considered as separate species according to the biological species concept. This demonstrates that cryptic species within parasitoid Hymenoptera also occur in Central Europe in close contact to humans.}, } @article {pmid31631529, year = {2020}, author = {Bi, J and Wang, YF}, title = {The effect of the endosymbiont Wolbachia on the behavior of insect hosts.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {846-858}, pmid = {31631529}, issn = {1744-7917}, support = {31672352//National Natural Science Foundation of China/ ; 2017AHB050//International Cooperation Projects of Science and Technology of Hubei Province, China/ ; 2017AHB050//the International Cooperation Projects of Science and Technology of Hubei Province, China/ ; }, mesh = {Aggression ; Animals ; *Behavior, Animal ; Feeding Behavior ; Insecta/*microbiology/*physiology ; Learning ; Memory ; Sexual Behavior, Animal ; Sleep ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {As one of the most successful intracellular symbiotic bacteria, Wolbachia can infect many arthropods and nematodes. Wolbachia infection usually affects the reproduction of their hosts to promote their own proliferation and transmission. Currently, most of the studies focus on the mechanisms of Wolbachia interactions with host reproduction. However, in addition to distribution in the reproductive tissues, Wolbachia also infect various somatic tissues of their hosts, including the brain. This raises the potential that Wolbachia may influence some somatic processes, such as behaviors in their hosts. So far, information about the effects of Wolbachia infection on host behavior is still very limited. The present review presents the current literature on different aspects of the influence of Wolbachia on various behaviors, including sleep, learning and memory, mating, feeding and aggression in their insect hosts. We then highlight ongoing scientific efforts in the field that need addressing to advance this field, which can have significant implications for further developing Wolbachia as environmentally friendly biocontrol agents to control insect-borne diseases and agricultural pests.}, } @article {pmid31622546, year = {2020}, author = {Galis, F and van Alphen, JJM}, title = {Parthenogenesis and developmental constraints.}, journal = {Evolution & development}, volume = {22}, number = {1-2}, pages = {205-217}, doi = {10.1111/ede.12324}, pmid = {31622546}, issn = {1525-142X}, mesh = {Animals ; *Biological Evolution ; Invertebrates/*growth & development ; *Parthenogenesis ; Vertebrates/*growth & development ; }, abstract = {The absence of a paternal contribution in an unfertilized ovum presents two developmental constraints against the evolution of parthenogenesis. We discuss the constraint caused by the absence of a centrosome and the one caused by the missing set of chromosomes and how they have been broken in specific taxa. They are examples of only a few well-underpinned examples of developmental constraints acting at macro-evolutionary scales in animals. Breaking of the constraint of the missing chromosomes is the best understood and generally involves rare occasions of drastic changes of meiosis. These drastic changes can be best explained by having been induced, or at least facilitated, by sudden cytological events (e.g., repeated rounds of hybridization, endosymbiont infections, and contagious infections). Once the genetic and developmental machinery is in place for regular or obligate parthenogenesis, shifts to other types of parthenogenesis can apparently rather easily evolve, for example, from facultative to obligate parthenogenesis, or from pseudoarrhenotoky to haplodiploidy. We argue that the combination of the two developmental constraints forms a near-absolute barrier against the gradual evolution from sporadic to obligate or regular facultative parthenogenesis, which can probably explain why the occurrence of the highly advantageous mode of regular facultative parthenogenesis is so rare and entirely absent in vertebrates.}, } @article {pmid31620116, year = {2019}, author = {Li, F and Hua, H and Ali, A and Hou, M}, title = {Characterization of a Bacterial Symbiont Asaia sp. in the White-Backed Planthopper, Sogatella furcifera, and Its Effects on Host Fitness.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {2179}, pmid = {31620116}, issn = {1664-302X}, abstract = {The white-backed planthopper (WBPH), Sogatella furcifera Horváth (Hemiptera: Delphacidae), is an economically significant rice insect pest that harbors a primary fungal yeast-like symbiont (YLS), and some secondary bacterial symbionts like Wolbachia and Cardinium. In the present study, an additional bacterial symbiont in WBPH was characterized. Phylogenetic analysis employing the 16S rRNA gene showed a bacterium closely related to Asaia of Nilaparvata lugens and Nysius expressus, and Asaia krungthepensis. TEM observation of the bacterium showed the typical morphology of Asaia sp. with signature filamentous structures in the nucleoid region. These results indicate that the bacterium belongs to Asaia. The Asaia bacterium was detected in all the tested individual adults and tissues of the laboratory WBPH population but showed varying infection rates (ca 45%) in the field collected WBPH populations. Quantitative PCR analysis revealed that Asaia sp. were significantly more abundant in WBPH females than males, and mainly distributed in the guts, fatty bodies, and salivary glands. Asaia-infected WBPH were of shorter nymphal duration and heavier adult weight than Asaia-free WBPH, while Asaia-free WBPH comparatively fed more, indicating that Asaia plays a role in improving WBPH fitness through involvement in host's nutrient supply.}, } @article {pmid31617302, year = {2020}, author = {Biwot, JC and Zhang, HB and Liu, C and Qiao, JX and Yu, XQ and Wang, YF}, title = {Wolbachia-induced expression of kenny gene in testes affects male fertility in Drosophila melanogaster.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {869-882}, doi = {10.1111/1744-7917.12730}, pmid = {31617302}, issn = {1744-7917}, support = {31672352//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Drosophila Proteins/*genetics/metabolism ; Drosophila melanogaster/genetics/microbiology/*physiology ; Fertility/*genetics ; Gene Expression ; I-kappa B Kinase/*genetics/metabolism ; Male ; Testis/metabolism ; *Up-Regulation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are Gram-negative endosymbionts that are known to cause embryonic lethality when infected male insects mate with uninfected females or with females carrying a different strain of Wolbachia, a situation characterized as cytoplasmic incompatibility (CI). However, the mechanism of CI is not yet fully understood, although recent studies on Drosophila melanogaster have achieved great progress. Here, we found that Wolbachia infection caused changes in the expressions of several immunity-related genes, including significant upregulation of kenny (key), in the testes of D. melanogaster. Overexpression of key in fly testes led to a significant decrease in egg hatch rates when these flies mate with wild-type females. Wolbachia-infected females could rescue this embryonic lethality. Furthermore, in key overexpressing testes terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling signal was significantly stronger than in the control testes, and the level of reactive oxygen species was significantly increased. Overexpression of key also resulted in alterations of some other immunity-related gene expressions, including the downregulation of Zn72D. Knockdown of Zn72D in fly testes also led to a significant decrease in egg hatch rates. These results suggest that Wolbachia might induce the defect in male host fertility by immunity-related pathways and thus cause an oxidative damage and cell death in male testes.}, } @article {pmid31615889, year = {2019}, author = {Chen, H and Ronau, JA and Beckmann, JF and Hochstrasser, M}, title = {A Wolbachia nuclease and its binding partner provide a distinct mechanism for cytoplasmic incompatibility.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {116}, number = {44}, pages = {22314-22321}, pmid = {31615889}, issn = {1091-6490}, support = {R01 GM046904/GM/NIGMS NIH HHS/United States ; R01 GM053756/GM/NIGMS NIH HHS/United States ; T32 GM007223/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics/*metabolism ; Deoxyribonucleases/genetics/*metabolism ; Drosophila melanogaster/*microbiology/physiology ; *Host-Pathogen Interactions ; Infertility, Male/*microbiology ; Male ; Operon ; Pest Control, Biological ; Protein Binding ; Wolbachia/enzymology/genetics/*pathogenicity ; }, abstract = {Wolbachia are endosymbiotic bacteria that infect nearly half of all arthropod species. This pandemic is due in part to their ability to increase their transmission through the female germline, most commonly by a mechanism called cytoplasmic incompatibility (CI). The Wolbachia cid operon, encoding 2 proteins, CidA and CidB, the latter a deubiquitylating enzyme (DUB), recapitulates CI in transgenic Drosophila melanogaster However, some CI-inducing Wolbachia strains lack a DUB-encoding cid operon; it was therefore proposed that the related cin operon codes for an alternative CI system. Here we show that the Wolbachia cin operon encodes a nuclease, CinB, and a second protein, CinA, that tightly binds CinB. Recombinant CinB has nuclease activity against both single-stranded and double-stranded DNA but not RNA under the conditions tested. Expression of the cin operon in transgenic male flies induces male sterility and embryonic defects typical of CI. Importantly, transgenic CinA can rescue defects in egg-hatch rates when expressed in females. Expression of CinA also rescues CinB-induced growth defects in yeast. CinB has 2 PD-(D/E)xK nuclease domains, and both are required for nuclease activity and for toxicity in yeast and flies. Our data suggest a distinct mechanism for CI involving a nuclease toxin and highlight the central role of toxin-antidote operons in Wolbachia-induced cytoplasmic incompatibility.}, } @article {pmid31615544, year = {2019}, author = {Ghosh, A and Jasperson, D and Cohnstaedt, LW and Brelsfoard, CL}, title = {Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {483}, pmid = {31615544}, issn = {1756-3305}, support = {Start up funds//Texas Tech University/ ; }, mesh = {Aedes/cytology ; Animals ; Biological Control Agents ; Cell Line/microbiology ; Ceratopogonidae/immunology/*microbiology ; Immunity/genetics ; In Situ Hybridization, Fluorescence ; Insect Vectors/immunology/*microbiology ; Pest Control, Biological/methods ; Phenotype ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Reproduction ; Transfection/*methods ; Wolbachia/genetics/immunology/*pathogenicity ; }, abstract = {BACKGROUND: Biting midges of the genus Culicoides vector multiple veterinary pathogens and are difficult to control. Endosymbionts particularly Wolbachia pipientis may offer an alternative to control populations of Culicoides and/or impact disease transmission in the form of population suppression or replacement strategies.

METHODS: Culicoides sonorensis cell lines were transfected with a Wolbachia infection using a modified shell vial technique. Infections were confirmed using PCR and cell localization using fluorescent in situ hybridization (FISH). The stability of Wolbachia infections and density was determined by qPCR. qPCR was also used to examine immune genes in the IMD, Toll and JACK/STAT pathways to determine if Wolbachia were associated with an immune response in infected cells.

RESULTS: Here we have transfected two Culicoides sonorensis cell lines (W3 and W8) with a Wolbachia infection (walbB) from donor Aedes albopictus Aa23 cells. PCR and FISH showed the presence of Wolbachia infections in both C. sonorensis cell lines. Infection densities were higher in the W8 cell lines when compared to W3. In stably infected cells, genes in the immune Toll, IMD and JAK/STAT pathways were upregulated, along with Attacin and an Attacin-like anti-microbial peptides.

CONCLUSIONS: The successful introduction of Wolbachia infections in C. sonorensis cell lines and the upregulation of immune genes, suggest the utility of using Wolbachia for a population replacement and/or population suppression approach to limit the transmission of C. sonorensis vectored diseases. Results support the further investigation of Wolbachia induced pathogen inhibitory effects in Wolbachia-infected C. sonorensis cell lines and the introduction of Wolbachia into C. sonorensis adults via embryonic microinjection to examine for reproductive phenotypes and host fitness effects of a novel Wolbachia infection.}, } @article {pmid31608104, year = {2019}, author = {Tseng, SP and Wetterer, JK and Suarez, AV and Lee, CY and Yoshimura, T and Shoemaker, D and Yang, CS}, title = {Genetic Diversity and Wolbachia Infection Patterns in a Globally Distributed Invasive Ant.}, journal = {Frontiers in genetics}, volume = {10}, number = {}, pages = {838}, pmid = {31608104}, issn = {1664-8021}, abstract = {Understanding the phylogeographic history of an invasive species may facilitate reconstructing the history and routes of its invasion. The longhorn crazy ant, Paratrechina longicornis, is a ubiquitous agricultural and household pest throughout much of the tropics and subtropics, but little is known about the history of its spread. Here, we examine worldwide genetic variation in P. longicornis and its associated Wolbachia bacterial symbionts. Analyses of mtDNA sequences of 248 P. longicornis workers (one per colony) from 13 geographic regions reveal two highly diverged mtDNA clades that co-occur in most of the geographic regions. These two mtDNA clades are associated with different Wolbachia infection patterns, but are not congruent with patterns of nDNA (microsatellite) variation. Multilocus sequence typing reveals two distinct Wolbachia strains in P. longicornis, namely, wLonA and wLonF. The evolutionary histories of these two strains differ; wLonA appears to be primarily transmitted maternally, and patterns of mtDNA and nDNA variation and wLonA infection status are consistent with a relatively recent Wolbachia-induced selective sweep. In contrast, the observed patterns of mtDNA variation and wLonF infections suggest frequent horizontal transfer and losses of wLonF infections. The lack of nDNA structure among sampled geographic regions coupled with the finding that numerous mtDNA haplotypes are shared among regions implies that inadvertent long-distance movement through human commerce is common in P. longicornis and has shaped the genetic structure of this invasive ant worldwide.}, } @article {pmid31603613, year = {2020}, author = {Caputo, B and Moretti, R and Manica, M and Serini, P and Lampazzi, E and Bonanni, M and Fabbri, G and Pichler, V and Della Torre, A and Calvitti, M}, title = {A bacterium against the tiger: preliminary evidence of fertility reduction after release of Aedes albopictus males with manipulated Wolbachia infection in an Italian urban area.}, journal = {Pest management science}, volume = {76}, number = {4}, pages = {1324-1332}, doi = {10.1002/ps.5643}, pmid = {31603613}, issn = {1526-4998}, mesh = {*Aedes ; Animals ; Female ; Fertility ; Italy ; Male ; Mosquito Control ; *Wolbachia ; }, abstract = {BACKGROUND: Novel tools are needed to reduce the nuisance and risk of exotic arbovirus transmission associated with the colonization of temperate regions by Aedes albopictus. The incompatible insect technique (IIT) is a population suppression approach based on cytoplasmic incompatibility between males with manipulated endosymbionts and wild females. Here, we present the results of the first field experiment in Europe to assess the capacity of an Ae. albopictus line (ARwP) deprived of its natural endosymbiont Wolbachia and transinfected with a Wolbachia strain from the mosquito Culex pipiens, to sterilize wild females.

RESULTS: We released ∼ 4500 ARwP males weekly for 6 weeks in a green area within urban Rome (Italy) and carried out egg (N = 13 442), female (N = 128) and male (N = 352) collections. Egg (N = 13 783) and female (N = 48) collections were also carried out at two untreated control sites. The percentage of viable eggs during release was, on average, significantly lower in treated sites than in control sites, with the greatest difference (16%) seen after the fourth release. The ARwP to wild male ratio in the release spots between day 3 after the first ARwP male release and day 7 after the last release was, on average, 7:10. Released males survived up to 2 weeks. Approximately 30% of females collected in the release spots showed 100% sterility and 20% showed strongly reduced fertility compared with control sites.

CONCLUSIONS: Results support the potential of IIT as a tool contributing to Ae. albopictus control in the urban context, and stress the need for larger field trials to evaluate the cost-efficacy of the approach in suppressing wild populations. © 2019 Society of Chemical Industry.}, } @article {pmid31600190, year = {2019}, author = {Becking, T and Chebbi, MA and Giraud, I and Moumen, B and Laverré, T and Caubet, Y and Peccoud, J and Gilbert, C and Cordaux, R}, title = {Sex chromosomes control vertical transmission of feminizing Wolbachia symbionts in an isopod.}, journal = {PLoS biology}, volume = {17}, number = {10}, pages = {e3000438}, pmid = {31600190}, issn = {1545-7885}, mesh = {Alleles ; Animals ; Female ; Genotype ; Homozygote ; Isopoda/*genetics/microbiology ; Male ; Models, Genetic ; Quantitative Trait, Heritable ; *Sex Chromosomes ; *Sex Determination Processes ; Sex Ratio ; Symbiosis/*genetics ; Wolbachia/*physiology ; }, abstract = {Microbial endosymbiosis is widespread in animals, with major ecological and evolutionary implications. Successful symbiosis relies on efficient vertical transmission through host generations. However, when symbionts negatively affect host fitness, hosts are expected to evolve suppression of symbiont effects or transmission. Here, we show that sex chromosomes control vertical transmission of feminizing Wolbachia endosymbionts in the isopod Armadillidium nasatum. Theory predicts that the invasion of an XY/XX species by cytoplasmic sex ratio distorters is unlikely because it leads to fixation of the unusual (and often lethal or infertile) YY genotype. We demonstrate that A. nasatum X and Y sex chromosomes are genetically highly similar and that YY individuals are viable and fertile, thereby enabling Wolbachia spread in this XY-XX species. Nevertheless, we show that Wolbachia cannot drive fixation of YY individuals, because infected YY females do not transmit Wolbachia to their offspring, unlike XX and XY females. The genetic basis fits the model of a Y-linked recessive allele (associated with an X-linked dominant allele), in which the homozygous state suppresses Wolbachia transmission. Moreover, production of all-male progenies by infected YY females restores a balanced sex ratio at the host population level. This suggests that blocking of Wolbachia transmission by YY females may have evolved to suppress feminization, thereby offering a whole new perspective on the evolutionary interplay between microbial symbionts and host sex chromosomes.}, } @article {pmid31599711, year = {2019}, author = {Nainu, F and Trenerry, A and Johnson, KN}, title = {Wolbachia-mediated antiviral protection is cell-autonomous.}, journal = {The Journal of general virology}, volume = {100}, number = {11}, pages = {1587-1592}, doi = {10.1099/jgv.0.001342}, pmid = {31599711}, issn = {1465-2099}, mesh = {Animals ; Cell Line ; Drosophila melanogaster/*microbiology/*virology ; *Host Microbial Interactions ; *Microbial Interactions ; Viruses/*growth & development ; Wolbachia/*growth & development ; }, abstract = {Vector-borne viral diseases pose significant risks to human health. To control the transmission of these viruses, a number of approaches are required. The ability of the intracellular bacteria Wolbachia to limit viral accumulation and transmission in some arthropod hosts, highlights its potential as a biocontrol agent. Whilst Wolbachia can reduce the transmission of several epidemiologically important viruses, protection is not consistent amongst all insects, viruses and strains of Wolbachia, which confounds elucidation of the mechanisms that underly this protection. Evidence of different mechanisms has emerged, but is not always consistent, suggesting the tripartite interaction may be complex. Here we provide evidence that Wolbachia-mediated antiviral protection is dependent on the presence of Wolbachia in individual cells, and cannot be conferred to surrounding cells. Our results suggest that protection is cell-autonomous, and this has several mechanistic implications, which can direct future research.}, } @article {pmid31596462, year = {2019}, author = {Wang, X and Xiong, X and Cao, W and Zhang, C and Werren, JH and Wang, X}, title = {Genome Assembly of the A-Group Wolbachia in Nasonia oneida Using Linked-Reads Technology.}, journal = {Genome biology and evolution}, volume = {11}, number = {10}, pages = {3008-3013}, pmid = {31596462}, issn = {1759-6653}, mesh = {Animals ; *Genome, Bacterial ; Genomics/*methods ; Multilocus Sequence Typing ; Wasps/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia are obligate intracellular bacteria which commonly infect various nematode and arthropod species. Genome sequences have been generated from arthropod samples following enrichment for the intracellular bacteria, and genomes have also been assembled from arthropod whole-genome sequencing projects. However, these methods remain challenging for infections that occur at low titers in hosts. Here we report the first Wolbachia genome assembled from host sequences using 10× Genomics linked-reads technology. The high read depth attainable by this method allows for recovery of intracellular bacteria that are at low concentrations. Based on the depth differences (714× for the insect and 59× for the bacterium), we assembled the genome of a Wolbachia in the parasitoid jewel wasp species Nasonia oneida. The final draft assembly consists of 1,293, 06 bp in 47 scaffolds with 1,114 coding genes and 97.01% genome completeness assessed by checkM. Comparisons of the five Multi Locus Sequence Typing genes revealed that the sequenced Wolbachia genome is the A1 strain (henceforth wOneA1) previously reported in N. oneida. Pyrosequencing confirms that the wasp strain lacks A2 and B types previously detected in this insect, which were likely lost during laboratory culturing. Assembling bacterial genomes from host genome projects can provide an effective method for sequencing bacterial genomes, even when the infections occur at low density in sampled tissues.}, } @article {pmid31596027, year = {2020}, author = {Bing, XL and Lu, YJ and Xia, CB and Xia, X and Hong, XY}, title = {Transcriptome of Tetranychus urticae embryos reveals insights into Wolbachia-induced cytoplasmic incompatibility.}, journal = {Insect molecular biology}, volume = {29}, number = {2}, pages = {193-204}, doi = {10.1111/imb.12620}, pmid = {31596027}, issn = {1365-2583}, mesh = {Animals ; Cytoplasm ; Embryo, Nonmammalian/microbiology/physiology ; Female ; Male ; Reproduction ; Tetranychidae/embryology/growth & development/microbiology/*physiology ; *Transcriptome ; Wolbachia/*physiology ; }, abstract = {The endosymbiont Wolbachia is known for manipulating host reproduction in selfish ways. However, the molecular mechanisms have not yet been investigated in embryos. Here, we found that Wolbachia had no effect on the number of deposited eggs in Tetranychus urticae Koch (Acari: Tetranychidae) but caused two types of reproductive manipulation: killing uninfected female embryos via cytoplasmic incompatibility (CI) and increasing the hatching ratio of infected female embryos. RNA sequencing analyses showed that 145 genes were differentially expressed between Wolbachia-infected (WI) and Wolbachia-uninfected (WU) embryos. Wolbachia infection down-regulated messenger RNA (mRNA) expression of glutathione S-transferase that could buffer oxidative stress. In addition, 1613 and 294 genes were identified as CI-specific up-/down-regulated genes. Compared to WU and WI embryos, embryos of CI cross strongly expressed genes involved in transcription, translation, tissue morphogenesis, DNA damage and mRNA surveillance. In contrast, most of the genes associated with energy production and metabolism were down-regulated in the CI embryos compared to the WU and WI embryos, which provides some clues as to the cause of death of CI embryos. These results identify several genes that could be candidates for explaining Wolbachia-induced CI. Our data form a basis to help elucidate the molecular consequences of CI in embryos.}, } @article {pmid31595328, year = {2020}, author = {Li, F and Li, P and Hua, H and Hou, M and Wang, F}, title = {Diversity, Tissue Localization, and Infection Pattern of Bacterial Symbionts of the White-Backed Planthopper, Sogatella furcifera (Hemiptera: Delphacidae).}, journal = {Microbial ecology}, volume = {79}, number = {3}, pages = {720-730}, pmid = {31595328}, issn = {1432-184X}, support = {2018YFD0200300//National Key R&D Project of China/ ; 31371951//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Bacteria/classification ; DNA, Bacterial/analysis ; Female ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Microbiota/*physiology ; RNA, Ribosomal, 16S/analysis ; Symbiosis ; }, abstract = {The white-backed planthopper (WBPH), Sogatella furcifera (Horváth), is a destructive pest of rice. Bacterial symbionts play an important role in insect hosts, especially hemipteran hosts. This study was designed to examine the bacterial symbionts of the WBPH using 16S rDNA high-throughput sequencing. A total of 63 and 177 operational taxonomic units (OTUs) were identified in females and males of three WBPH populations, respectively. These OTUs included bacteria of 75 genera from 11 phyla, where Wolbachia, Cardinium, and Asaia were the dominant genera, accounting for over 97.99% of all the symbiotic bacteria. Fluorescence in situ hybridization detected Wolbachia, Cardinium, and Asaia in the salivary glands, guts, testes, and eggs of the WBPH, indicating the potential for both horizontal and vertical transmission. Moreover, the infection pattern of the three dominant bacterial symbionts was detected in six WBPH populations. The frequencies of Wolbachia infection of females and Cardinium infection of both sexes were over 96.7%. Wolbachia infection of males ranged between 46.7 and 63.3%, which was significantly lower than that observed for females. Asaia infection of both sexes varied substantially among the populations. These results indicate that the complex host-symbiotic bacteria interaction is influenced by host sex and geographical origin and potentially by the transmission modes of the symbionts.}, } @article {pmid31592190, year = {2019}, author = {Reynolds, LA and Hornett, EA and Jiggins, CD and Hurst, GDD}, title = {Suppression of Wolbachia-mediated male-killing in the butterfly Hypolimnas bolina involves a single genomic region.}, journal = {PeerJ}, volume = {7}, number = {}, pages = {e7677}, pmid = {31592190}, issn = {2167-8359}, abstract = {BACKGROUND: Sex ratio distorting agents (maternally inherited symbionts and meiotically-driving sex chromosomes) are common in insects. When these agents rise to high frequencies they create strong population sex ratio bias and selection then favours mutations that act to restore the rare sex. Despite this strong selection pressure, the evolution of mutations that suppress sex ratio distorting elements appears to be constrained in many cases, where sex-biased populations persist for many generations. This scenario has been observed in the butterfly Hypolimnas bolina, where Wolbachia-mediated male killing endured for 800-1,000 generations across multiple populations before the evolution of suppression. Here we test the hypothesis that this evolutionary lag is the result of suppression being a multilocus trait requiring multiple mutations.

METHODS: We developed genetic markers, based on conservation of synteny, for each H. bolina chromosome and verified coverage using recombinational mapping. We then used a Wolbachia-infected mapping family to assess each chromosome for the presence of loci required for male survival, as determined by the presence of markers in all surviving sons.

RESULTS: Informative markers were obtained for each of the 31 chromosomes in H. bolina. The only marker that cosegregated with suppression was located on chromosome 25. A genomic region necessary for suppression has previously been located on this chromosome. We therefore conclude that a single genomic region of the H. bolina genome is necessary for male-killing suppression.

DISCUSSION: The evolutionary lag observed in our system is not caused by a need for changes at multiple genomic locations. The findings favour hypotheses in which either multiple mutations are required within a single genomic region, or the suppressor mutation is a singularly rare event.}, } @article {pmid31568486, year = {2019}, author = {Voronin, D and Schnall, E and Grote, A and Jawahar, S and Ali, W and Unnasch, TR and Ghedin, E and Lustigman, S}, title = {Pyruvate produced by Brugia spp. via glycolysis is essential for maintaining the mutualistic association between the parasite and its endosymbiont, Wolbachia.}, journal = {PLoS pathogens}, volume = {15}, number = {9}, pages = {e1008085}, pmid = {31568486}, issn = {1553-7374}, support = {T32 AI007180/AI/NIAID NIH HHS/United States ; F31 AI131527/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia/genetics/*metabolism/*microbiology ; Brugia malayi/genetics/metabolism/microbiology ; Brugia pahangi/genetics/metabolism/microbiology ; Female ; Filariasis/metabolism/microbiology/parasitology ; Genes, Helminth ; Glycolysis ; Host Microbial Interactions ; Host-Parasite Interactions ; Humans ; Male ; Pyruvic Acid/*metabolism ; Symbiosis ; Wolbachia/genetics/*metabolism ; }, abstract = {Human parasitic nematodes are the causative agents of lymphatic filariasis (elephantiasis) and onchocerciasis (river blindness), diseases that are endemic to more than 80 countries and that consistently rank in the top ten for the highest number of years lived with disability. These filarial nematodes have evolved an obligate mutualistic association with an intracellular bacterium, Wolbachia, a symbiont that is essential for the successful development, reproduction, and survival of adult filarial worms. Elimination of the bacteria causes adult worms to die, making Wolbachia a primary target for developing new interventional tools to combat filariases. To further explore Wolbachia as a promising indirect macrofilaricidal drug target, the essential cellular processes that define the symbiotic Wolbachia-host interactions need to be identified. Genomic analyses revealed that while filarial nematodes encode all the enzymes necessary for glycolysis, Wolbachia does not encode the genes for three glycolytic enzymes: hexokinase, 6-phosphofructokinase, and pyruvate kinase. These enzymes are necessary for converting glucose into pyruvate. Wolbachia, however, has the full complement of genes required for gluconeogenesis starting with pyruvate, and for energy metabolism via the tricarboxylic acid cycle. Therefore, we hypothesized that Wolbachia might depend on host glycolysis to maintain a mutualistic association with their parasitic host. We did conditional experiments in vitro that confirmed that glycolysis and its end-product, pyruvate, sustain this symbiotic relationship. Analysis of alternative sources of pyruvate within the worm indicated that the filarial lactate dehydrogenase could also regulate the local intracellular concentration of pyruvate in proximity to Wolbachia and thus help control bacterial growth via molecular interactions with the bacteria. Lastly, we have shown that the parasite's pyruvate kinase, the enzyme that performs the last step in glycolysis, could be a potential novel anti-filarial drug target. Establishing that glycolysis is an essential component of symbiosis in filarial worms could have a broader impact on research focused on other intracellular bacteria-host interactions where the role of glycolysis in supporting intracellular survival of bacteria has been reported.}, } @article {pmid31568480, year = {2019}, author = {Gawande, SJ and Anandhan, S and Ingle, A and Roylawar, P and Khandagale, K and Gawai, T and Jacobson, A and Asokan, R and Singh, M}, title = {Microbiome profiling of the onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae).}, journal = {PloS one}, volume = {14}, number = {9}, pages = {e0223281}, pmid = {31568480}, issn = {1932-6203}, mesh = {Actinobacteria/classification/*genetics/isolation & purification ; Animals ; Bacterial Typing Techniques ; Bacteroidetes/classification/*genetics/isolation & purification ; Cyanobacteria/classification/*genetics/isolation & purification ; Firmicutes/classification/*genetics/isolation & purification ; Gastrointestinal Microbiome/*genetics ; Genetic Variation ; India ; Phylogeny ; Proteobacteria/classification/*genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Symbiosis/genetics ; Thysanoptera/*microbiology ; Nicotiana/parasitology ; Wolbachia/classification/genetics/isolation & purification ; }, abstract = {The gut microbial community structure of adult Thrips tabaci collected from 10 different agro-climatically diverse locations of India was characterized by using the Illumina MiSeq platform to amplify the V3 region of the 16S rRNA gene of bacteria present in the sampled insects. Analyses were performed to study the bacterial communities associated with Thrips tabaci in India. The complete bacterial metagenome of T. tabaci was comprised of 1662 OTUs of which 62.25% belong to known and 37.7% of unidentified/unknown bacteria. These OTUs constituted 21 bacterial phyla of 276 identified genera. Phylum Proteobacteria was predominant, followed by Actinobacteria, Firmicutes, Bacteroidetes and Cyanobacteria. Additionally, the occurrence of the reproductive endosymbiont, Wolbachia was detected at two locations (0.56%) of the total known OTUs. There is high variation in diversity and species richness among the different locations. Alpha-diversity metrics indicated the higher gut bacterial diversity at Bangalore and lowest at Rahuri whereas higher bacterial species richness at T. tabaci samples from Imphal and lowest at Jhalawar. Beta diversity analyses comparing bacterial communities between the samples showed distinct differences in bacterial community composition of T. tabaci samples from different locations. This paper also constitutes the first record of detailed bacterial communities associated with T. tabaci. The location-wise variation in microbial metagenome profile of T. tabaci suggests that bacterial diversity might be governed by its population genetic structure, environment and habitat.}, } @article {pmid31566662, year = {2019}, author = {Detcharoen, M and Arthofer, W and Schlick-Steiner, BC and Steiner, FM}, title = {Wolbachia megadiversity: 99% of these microorganismic manipulators unknown.}, journal = {FEMS microbiology ecology}, volume = {95}, number = {11}, pages = {}, doi = {10.1093/femsec/fiz151}, pmid = {31566662}, issn = {1574-6941}, mesh = {Animals ; Arthropods/microbiology ; *Biodiversity ; Ecosystem ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia (Alphaproteobacteria) are the most widespread endosymbionts of arthropods, manipulating their hosts by various means to maximize the number of host individuals infected. Based on quantitative analyzes of the published literature from Web of Science® and of DNA sequences of arthropod-hosted Wolbachia from GenBank, we made plausible that less than 1% of the expected 100 000 strains of Wolbachia in arthropods is known. Our findings suggest that more and globally better coordinated efforts in screening arthropods are needed to explore the true Wolbachia diversity and to help us understand the ecology and evolution of these host-endosymbiont interactions.}, } @article {pmid31565105, year = {2019}, author = {Huang, K and Kelly, PJ and Zhang, J and Yang, Y and Liu, W and Kalalah, A and Wang, C}, title = {Molecular Detection of Bartonella spp. in China and St. Kitts.}, journal = {The Canadian journal of infectious diseases & medical microbiology = Journal canadien des maladies infectieuses et de la microbiologie medicale}, volume = {2019}, number = {}, pages = {3209013}, pmid = {31565105}, issn = {1712-9532}, abstract = {Bartonella are vector-borne hemotropic bacteria that infect a wide variety of hosts, including people. While there are PCR assays that can identify individual or groups of Bartonella, there is no reliable molecular method to simultaneously detect all species while maintaining genus specificity and sensitivity. By comparing highly conserved 16S rRNA sequences of the better-recognized Bartonella spp. on GenBank, we selected primers and probes for a genus-specific pan-Bartonella FRET-qPCR. Then, a gltA-based Bartonella PCR was established by selecting primers for a highly variable region of gltA, of which the sequenced amplicons could identify individual Bartonella spp. The pan-Bartonella FRET-qPCR did not detect negative controls (Brucella spp., Anaplasma spp., Rickettsia spp., Coxiella burnetii, and Wolbachia) but reliably detected as few as two copies of the positive control (Bartonella henselae) per reaction. There was complete agreement between the pan-Bartonella FRET-qPCR and the gltA-based Bartonella PCR in detecting Bartonella in convenience test samples from China and St. Kitts: cats (26%; 81/310), Ctenocephalides felis (20%; 12/60), cattle (24%; 23/98), and donkeys (4%; 1/20). Sequencing of the gltA-based Bartonella PCR products revealed B. henselae (70%; 57/81) and B. clarridgeiae (30%; 24/81) in cats and C. felis (67%; 8/12, and 33%; 4/12, respectively) and B. bovis in cattle (23.5%; 23/98) and donkeys (4.0%; 1/24). The pan-Bartonella FRET-qPCR and gltA-based Bartonella PCR we developed are highly sensitive and specific in detecting recognized Bartonella spp. in a single reaction. The pan-Bartonella FRET-qPCR is convenient requiring no gel electrophoresis and providing copy numbers, while the gltA-based Bartonella PCR reliably differentiates individual Bartonella species. The use of these PCRs should greatly facilitate large-scale surveillance studies and the diagnosis of infections in clinical samples.}, } @article {pmid31557154, year = {2019}, author = {Andersen, BJ and Rosa, BA and Kupritz, J and Meite, A and Serge, T and Hertz, MI and Curtis, K and King, CL and Mitreva, M and Fischer, PU and Weil, GJ}, title = {Systems analysis-based assessment of post-treatment adverse events in lymphatic filariasis.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {9}, pages = {e0007697}, pmid = {31557154}, issn = {1935-2735}, mesh = {Adolescent ; Adult ; Aged ; Albendazole/administration & dosage/adverse effects ; Antigens, Helminth/blood ; Cytokines/blood/immunology ; Diethylcarbamazine/adverse effects/therapeutic use ; Elephantiasis, Filarial/*drug therapy/genetics/immunology ; Female ; Filaricides/adverse effects/*therapeutic use ; Humans ; Ivermectin/administration & dosage/adverse effects ; Male ; Middle Aged ; Young Adult ; }, abstract = {BACKGROUND: Lymphatic filariasis (LF) is a neglected tropical disease, and the Global Program to Eliminate LF delivers mass drug administration (MDA) to 500 million people every year. Adverse events (AEs) are common after LF treatment.

To better understand the pathogenesis of AEs, we studied LF-patients from a treatment trial. Plasma levels of many filarial antigens increased post-treatment in individuals with AEs, and this is consistent with parasite death. Circulating immune complexes were not elevated in these participants, and the classical complement cascade was not activated. Multiple cytokines increased after treatment in persons with AEs. A transcriptomic analysis was performed for nine individuals with moderate systemic AEs and nine matched controls. Differential gene expression analysis identified a significant transcriptional signature associated with post-treatment AEs; 744 genes were upregulated. The transcriptional signature was enriched for TLR and NF-κB signaling. Increased expression of seven out of the top eight genes upregulated in persons with AEs were validated by qRT-PCR, including TLR2.

CONCLUSIONS/SIGNIFICANCE: This is the first global study of changes in gene expression associated with AEs after treatment of lymphatic filariasis. Changes in cytokines were consistent with prior studies and with the RNAseq data. These results suggest that Wolbachia lipoprotein is involved in AE development, because it activates TLR2-TLR6 and downstream NF-κB. Additionally, LPS Binding Protein (LBP, which shuttles lipoproteins to TLR2) increased post-treatment in individuals with AEs. Improved understanding of the pathogenesis of AEs may lead to improved management, increased MDA compliance, and accelerated LF elimination.}, } @article {pmid31550397, year = {2019}, author = {Bonneau, M and Caputo, B and Ligier, A and Caparros, R and Unal, S and Perriat-Sanguinet, M and Arnoldi, D and Sicard, M and Weill, M}, title = {Variation in Wolbachia cidB gene, but not cidA, is associated with cytoplasmic incompatibility mod phenotype diversity in Culex pipiens.}, journal = {Molecular ecology}, volume = {28}, number = {21}, pages = {4725-4736}, pmid = {31550397}, issn = {1365-294X}, mesh = {Animals ; Animals, Genetically Modified/genetics ; Crosses, Genetic ; Culex/*genetics/*microbiology ; Cytoplasm/*genetics/*microbiology ; Drosophila melanogaster/genetics/microbiology ; Female ; Genes, Bacterial/*genetics ; Italy ; Male ; Phenotype ; Reproduction/genetics ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Endosymbiotic Wolbachia bacteria are, to date, considered the most widespread symbionts in arthropods and are the cornerstone of major biological control strategies. Such a high prevalence is based on the ability of Wolbachia to manipulate their hosts' reproduction. One manipulation called cytoplasmic incompatibility (CI) is based on the death of the embryos generated by crosses between infected males and uninfected females or between individuals infected with incompatible Wolbachia strains. CI can be seen as a modification-rescue system (or mod-resc) in which paternal Wolbachia produce mod factors, inducing embryonic defects, unless the maternal Wolbachia produce compatible resc factors. Transgenic experiments in Drosophila melanogaster and Saccharomyces cerevisiae converged towards a model where the cidB Wolbachia gene is involved in the mod function while cidA is involved in the resc function. However, as cidA expression in Drosophila males was required to observe CI, it has been proposed that cidA could be involved in both resc and mod functions. A recent correlative study in natural Culex pipiens mosquito populations has revealed an association between specific cidA and cidB variations and changes in mod phenotype, also suggesting a role for both these genes in mod diversity. Here, by studying cidA and cidB genomic repertoires of individuals from newly sampled natural C. pipiens populations harbouring wPipIV strains from North Italy, we reinforce the link between cidB variation and mod phenotype variation fostering the involvement of cidB in the mod phenotype diversity. However, no association between any cidA variants or combination of cidA variants and mod phenotype variation was observed. Taken together our results in natural C. pipiens populations do not support the involvement of cidA in mod phenotype variation.}, } @article {pmid31550003, year = {2019}, author = {Avtzis, DN and Schebeck, M and Petsopoulos, D and Memtsas, GI and Stauffer, C and Kavallieratos, NG and Athanassiou, CG and Boukouvala, MC}, title = {New Data on the Range Expansion of the Thaumetopoea pityocampa (Lepidoptera: Notodontidae) 'ENA clade' in Greece: The Role of Bacterial Endosymbionts.}, journal = {Journal of economic entomology}, volume = {112}, number = {6}, pages = {2761-2766}, doi = {10.1093/jee/toz216}, pmid = {31550003}, issn = {1938-291X}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Greece ; Humans ; *Moths ; *Wolbachia ; }, abstract = {The pine processionary moth, Thaumetopoea pityocampa (Denis and Schiffermüller), is an important insect in the Mediterranean region, as it defoliates pines and its urticating hairs can cause allergic reactions in humans and animals. Moreover, this species exhibits an interesting genetic structure as recently a distinct East-North African mtDNA lineage ('ENA clade') has been described. This clade has been recently detected in Greek populations where it has currently expanded its range by replacing the 'endemic' T. pityocampa lineages. Here, we report new data on the rapid spread of 'ENA clade' in the Greek island Evoia in only a few years. As the underlying mechanisms of the 'ENA clade' range expansion has not been studied so far, we screened T. pityocampa for an infection with the heritable bacterial endosymbionts Wolbachia (Bacteria: Anaplasmataceae), Cardinium (Bacteria: Bacteroidaceae), Rickettsia (Bacteria: Rickettsiaceae) and Spiroplasma (Bacteria: Spiroplasmataceae). These bacteria can manipulate the reproduction of infected hosts, something that could potentially explain the rapid spread of 'ENA clade' lineage. Therefore, we screened 28 individuals that exhibited T. pityocampa 'ENA clade' and 'endemic' T. pityocampa haplotypes from nine populations scattered all over Greece. None of them was infected with any of the four endosymbionts, suggesting that these bacteria do not cause reproductive manipulations in T. pityocampa lineages and, thus, other factors should be explored in future research efforts.}, } @article {pmid31549736, year = {2020}, author = {Ogawa, M and Takahashi, M and Matsutani, M and Takada, N and Noda, S and Saijo, M}, title = {Obligate intracellular bacteria diversity in unfed Leptotrombidium scutellare larvae highlights novel bacterial endosymbionts of mites.}, journal = {Microbiology and immunology}, volume = {64}, number = {1}, pages = {1-9}, doi = {10.1111/1348-0421.12745}, pmid = {31549736}, issn = {1348-0421}, support = {//"OSIMO" Foundation (Japan)/ ; }, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; *Bacterial Physiological Phenomena ; *Biodiversity ; High-Throughput Nucleotide Sequencing ; Humans ; Larva/*microbiology ; Life Cycle Stages ; Mites/*microbiology ; Orientia tsutsugamushi ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsia/classification/genetics ; Scrub Typhus/microbiology ; Sequence Analysis ; *Symbiosis ; Trombiculidae/*microbiology ; Wolbachia/classification ; }, abstract = {It is well known that the mite Leptotrombidium scutellare carries the pathogen of scrub typhus, Orientia tsutsugamushi. However, our understanding of other bacterial endosymbionts of mites is limited. This study investigated the diversity of the obligate intracellular bacteria carried by L. scutellare using 16S rRNA gene amplicon analysis with next-generation sequencing. The results showed that the detected bacteria were classified into the genera Rickettsia, Wolbachia, and Rickettsiella and an unknown genus of the order Rickettsiales. For further classification of the detected bacteria, a representative read that was most closely related to the assigned taxonomic classification was subjected to homology search and phylogenic analysis. The results showed that some bacteria of the genus Rickettsia were identical or very close to the human pathogens Rickettsia akari, Rickettsia aeschlimannii, Rickettsia felis, and Rickettsia australis. The genetic distance between the genus Wolbachia bacteria in the present study and in previous reports is highly indicative that the bacteria in the present study can be classified as a new taxon of Wolbachia. This study detected obligate intracellular bacteria from unfed mites; thus, the mites did not acquire bacteria from infected animals or any other infectious sources. Finally, the present study demonstrated that various and novel bacterial endosymbionts of mites, in addition to O. tsutsugamushi, might uniquely evolve with the host mites throughout overlapping generations of the mite life cycle. The roles of the bacteria in mites and their pathogenicity should be further examined in studies based on bacterial isolation.}, } @article {pmid30596205, year = {2018}, author = {O'Neill, SL and Ryan, PA and Turley, AP and Wilson, G and Retzki, K and Iturbe-Ormaetxe, I and Dong, Y and Kenny, N and Paton, CJ and Ritchie, SA and Brown-Kenyon, J and Stanford, D and Wittmeier, N and Jewell, NP and Tanamas, SK and Anders, KL and Simmons, CP}, title = {Scaled deployment of Wolbachia to protect the community from dengue and other Aedes transmitted arboviruses.}, journal = {Gates open research}, volume = {2}, number = {}, pages = {36}, pmid = {30596205}, issn = {2572-4754}, support = {/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Background: A number of new technologies are under development for the control of mosquito transmitted viruses, such as dengue, chikungunya and Zika that all require the release of modified mosquitoes into the environment. None of these technologies has been able to demonstrate evidence that they can be implemented at a scale beyond small pilots. Here we report the first successful citywide scaled deployment of Wolbachia in the northern Australian city of Townsville. Methods: The wMel strain of Wolbachia was backcrossed into a local Aedes aegypti genotype and mass reared mosquitoes were deployed as eggs using mosquito release containers (MRCs). In initial stages these releases were undertaken by program staff but in later stages this was replaced by direct community release including the development of a school program that saw children undertake releases. Mosquito monitoring was undertaken with Biogents Sentinel (BGS) traps and individual mosquitoes were screened for the presence of Wolbachia with a Taqman qPCR or LAMP diagnostic assay. Dengue case notifications from Queensland Health Communicable Disease Branch were used to track dengue cases in the city before and after release. Results: Wolbachia was successfully established into local Ae. aegypti mosquitoes across 66 km [2] in four stages over 28 months with full community support. A feature of the program was the development of a scaled approach to community engagement. Wolbachia frequencies have remained stable since deployment and to date no local dengue transmission has been confirmed in any area of Townsville after Wolbachia has established, despite local transmission events every year for the prior 13 years and an epidemiological context of increasing imported cases. Conclusion: Deployment of Wolbachia into Ae. aegypti populations can be readily scaled to areas of ~60km [2] quickly and cost effectively and appears in this context to be effective at stopping local dengue transmission.}, } @article {pmid31534674, year = {2019}, author = {Sanaei, E and Husemann, M and Seiedy, M and Rethwisch, M and Tuda, M and Toshova, TB and Kim, MJ and Atanasova, D and Kim, I}, title = {Global genetic diversity, lineage distribution, and Wolbachia infection of the alfalfa weevil Hypera postica (Coleoptera: Curculionidae).}, journal = {Ecology and evolution}, volume = {9}, number = {17}, pages = {9546-9563}, pmid = {31534674}, issn = {2045-7758}, abstract = {The alfalfa weevil (Hypera postica) is a well-known example of a worldwide-distributed pest with high genetic variation. Based on the mitochondrial genes, the alfalfa weevil clusters into two main mitochondrial lineages. However, there is no clear picture of the global diversity and distribution of these lineages; neither the drivers of its diversification are known. However, it appears likely that historic demographic events including founder effects played a role. In addition, Wolbachia, a widespread intracellular parasite/symbiont, likely played an important role in the evolution of the species. Wolbachia infection so far was only detected in the Western lineage of H. postica with no information on the infecting strain, its frequency, and its consequences on the genetic diversity of the host. We here used a combination of mitochondrial and nuclear sequences of the host and sequence information on Wolbachia to document the distribution of strains and the degree of infection. The Eastern lineage has a higher genetic diversity and is found in the Mediterranean, the Middle East, Eastern Europe, and eastern America, whereas the less diverse Western lineage is found in Central Europe and the western America. Both lineages are infected with the same common strain of Wolbachia belonging to Supergroup B. Based on neutrality tests, selection tests, and the current distribution and diversification of Wolbachia in H. postica, we suggested the Wolbachia infection did not shape genetic diversity of the host. The introduced populations in the United States are generally genetically less diverse, which is in line with founder effects.}, } @article {pmid31530141, year = {2019}, author = {Dincă, V and Lee, KM and Vila, R and Mutanen, M}, title = {The conundrum of species delimitation: a genomic perspective on a mitogenetically super-variable butterfly.}, journal = {Proceedings. Biological sciences}, volume = {286}, number = {1911}, pages = {20191311}, pmid = {31530141}, issn = {1471-2954}, mesh = {Africa, Northern ; Animals ; Biological Evolution ; Butterflies/*genetics ; DNA, Mitochondrial/*analysis ; Europe ; Gene Flow ; *Genetic Speciation ; *Genome, Insect ; Kazakhstan ; *Phylogeny ; *Polymorphism, Single Nucleotide ; Restriction Mapping ; }, abstract = {The Palaearctic butterfly Melitaea didyma stands out as one of the most striking cases of intraspecific genetic differentiation detected in Lepidoptera: 11 partially sympatric mitochondrial lineages have been reported, displaying levels of divergence of up to 7.4%. To better understand the evolutionary processes underlying the diversity observed in mtDNA, we compared mtDNA and genome-wide SNP data using double-digest restriction site-associated DNA sequencing (ddRADseq) results from 93 specimens of M. didyma ranging from Morocco to eastern Kazakhstan. We found that, between ddRADseq and mtDNA results, there is a match only in populations that probably remained allopatric for long periods of time. Other mtDNA lineages may have resulted from introgression events and were probably affected by Wolbachia infection. The five main ddRADseq clades supported by STRUCTURE were parapatric or allopatric and showed high pairwise FST values, but some were also estimated to display various levels of gene flow. Melitaea didyma represents one of the first cases of deep mtDNA splits among European butterflies assessed by a genome-wide DNA analysis and reveals that the interpretation of patterns remains challenging even when a high amount of genomic data is available. These findings actualize the ongoing debate of species delimitation in allopatry, an issue probably of relevance to a significant proportion of global biodiversity.}, } @article {pmid31515862, year = {2019}, author = {Després, L}, title = {One, two or more species? Mitonuclear discordance and species delimitation.}, journal = {Molecular ecology}, volume = {28}, number = {17}, pages = {3845-3847}, doi = {10.1111/mec.15211}, pmid = {31515862}, issn = {1365-294X}, mesh = {Animals ; Butterflies/genetics ; Cell Nucleus/*genetics ; Climate ; DNA, Mitochondrial/genetics ; Gene Flow ; Geography ; High-Throughput Nucleotide Sequencing ; Mitochondria/*genetics ; *Phylogeny ; Polymorphism, Single Nucleotide/genetics ; Species Specificity ; }, abstract = {Delimiting species boundaries is central to understand ecological and evolutionary processes, and to monitor biodiversity patterns over time and space. Yet, most of our current knowledge on animal diversity and phylogeny relies on morphological and mitochondrial (mt) DNA variation, a popular molecular marker also used as a barcode to assign samples to species. For morphologically undistinguishable sympatric species (cryptic species), the congruence of several independent markers is necessary to define separate species. Nuclear markers are becoming more accessible, and have confirmed that cryptic species are widespread in all animal phyla (Fišer, Robinson, & Malard, 2018). However, striking differences between the mitochondrial and nuclear variation patterns are also commonly found within single species. Mitonuclear discordance can result from incomplete lineage sorting, sex-biased dispersal, asymmetrical introgression, natural selection or Wolbachia-mediated genetic sweeps. But more generally, the distinct mode of transmission of these two types of markers (maternal vs. biparental) is sufficient to explain their distinct sensitivity to purely demographic events such as spatial range and population size fluctuations over time. In a From the Cover manuscript in this issue of Molecular Ecology, Hijonosa et al. (2019) show that highly divergent mtDNA lineages coexist in a widespread European butterfly (Figure 1). None of the hundreds of nuclear markers analyzed was associated with mt lineages, nor was Wolbachia variation. These findings rule out the presence of cryptic species but shed light on complex demographic history of lineage divergence/fusion during the Pleistocene climatic fluctuations, and pave the way to a better integration of both mt and nuclear information in demographic models.}, } @article {pmid31513297, year = {2020}, author = {Schwartz, RA and Al-Qubati, Y and Zieleniewski, Ł and Shah, R and Kapila, R}, title = {Onchocerciasis (river blindness): larva-induced eczema (onchodermatitis) from an important oculocutaneous tropical disease spilling over into North America and Europe.}, journal = {International journal of dermatology}, volume = {59}, number = {9}, pages = {1065-1070}, doi = {10.1111/ijd.14614}, pmid = {31513297}, issn = {1365-4632}, mesh = {Animals ; *Eczema ; Europe ; Ivermectin/therapeutic use ; Larva ; North America ; *Onchocerciasis/complications/diagnosis/drug therapy ; *Onchocerciasis, Ocular/diagnosis/drug therapy/epidemiology ; }, abstract = {Onchocerciasis is a leading cause of blindness in the world. It may be seen in temperate climates of the United States and Europe in immigrants and travelers from endemic regions, often linked to poverty and war. One should be aware of an incubation period that can be up to 15 months. In its early stage and throughout its course, onchocerciasis has noteworthy skin findings, facilitating diagnosis, as onchodermatitis resembles common eczema with variable degrees of papular, lichenoid, atrophic, and pigmentary alterations, features not suggestive if one is unaware of an individual's immigration and travel history. The same concept applies for the encysted worms (onchocercomas), as they tend to appear as common skin cysts and benign neoplasms. New methods can be employed to increase diagnostic sensitivity and specificity. Ivermectin is the gold standard of therapy, the use of which has almost miraculously eliminated this disease from large areas of the earth. However, its effect remains isolated to microfilariae and can be devastating in those coinfected with Loa loa. Recently, the symbiotic relationship between adult worms and Wolbachia bacteria has been discovered and, with it, the possibility of adding doxycycline as a treatment option. We also discuss coinfection with HIV and other diseases.}, } @article {pmid31507561, year = {2019}, author = {Meng, L and Li, X and Cheng, X and Zhang, H}, title = {16S rRNA Gene Sequencing Reveals a Shift in the Microbiota of Diaphorina citri During the Psyllid Life Cycle.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {1948}, pmid = {31507561}, issn = {1664-302X}, abstract = {The Asian citrus psyllid (Diaphorina citri) is a major pest of citrus trees as it transmits Candidatus Liberibacter asiaticus (CLas). The composition of a host's microbiota can affect the evolution and ecological distribution of the host. This study monitored the compositional shifts in the citrus psyllid microbiota through all the life stages (egg, nymph 1-5 stages, and adult) by next-generation sequencing (NGS) and quantitative real-time PCR. There were clear differences in both α- and β-diversity of microbiota through the psyllid life stages. Microbiota diversity was markedly higher in the nymph 2-5 stages than in the adult, egg, and nymph 1 stages. Proteobacteria were dominant in all the life stages of D. citri, representing >97.5% of the total bacterial community, and Candidatus Profftella armature was the dominant genus in all the life stages. Data from the qPCR analysis showed an exponential increase in the populations of three D. citri endosymbionts: Candidatus Profftella armature, Candidatus Carsonella ruddii, and Wolbachia. The gut bacterium Pantoea was present in all the life stages, but it was markedly higher in the nymph 2-5 stages. The microbiota composition substantially differed among the egg-nymph 1, nymphs 2-5, and adult stages. Therefore, we successfully characterized the microbiota dynamics and thus identified a microbiota shift during the life cycle of D. citri by 16S rRNA gene sequencing and quantitative PCR. Moreover, 16S rRNA gene sequencing suggested that D. citri acquired the ability to bear CLas in the nymph 1 stage. This study enhances our understanding of microbial establishment in the developing D. citri and provides a reference resource for the identification of potential biocontrol approaches against this pest.}, } @article {pmid31505135, year = {2019}, author = {Ross, PA and Turelli, M and Hoffmann, AA}, title = {Evolutionary Ecology of Wolbachia Releases for Disease Control.}, journal = {Annual review of genetics}, volume = {53}, number = {}, pages = {93-116}, pmid = {31505135}, issn = {1545-2948}, support = {/WT_/Wellcome Trust/United Kingdom ; R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Communicable Disease Control/*methods ; Cytoplasm ; Environment ; Genetic Fitness ; Host-Pathogen Interactions/*physiology ; Insect Vectors/microbiology/*virology ; Insecta/microbiology/virology ; Mosquito Vectors/microbiology/virology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is an endosymbiotic Alphaproteobacteria that can suppress insect-borne diseases through decreasing host virus transmission (population replacement) or through decreasing host population density (population suppression). We contrast natural Wolbachia infections in insect populations with Wolbachia transinfections in mosquitoes to gain insights into factors potentially affecting the long-term success of Wolbachia releases. Natural Wolbachia infections can spread rapidly, whereas the slow spread of transinfections is governed by deleterious effects on host fitness and demographic factors. Cytoplasmic incompatibility (CI) generated by Wolbachia is central to both population replacement and suppression programs, but CI in nature can be variable and evolve, as can Wolbachia fitness effects and virus blocking. Wolbachia spread is also influenced by environmental factors that decrease Wolbachia titer and reduce maternal Wolbachia transmission frequency. More information is needed on the interactions between Wolbachia and host nuclear/mitochondrial genomes, the interaction between invasion success and local ecological factors, and the long-term stability of Wolbachia-mediated virus blocking.}, } @article {pmid31504075, year = {2019}, author = {Perlmutter, JI and Bordenstein, SR and Unckless, RL and LePage, DP and Metcalf, JA and Hill, T and Martinez, J and Jiggins, FM and Bordenstein, SR}, title = {The phage gene wmk is a candidate for male killing by a bacterial endosymbiont.}, journal = {PLoS pathogens}, volume = {15}, number = {9}, pages = {e1007936}, pmid = {31504075}, issn = {1553-7374}, support = {WT094664MA/WT_/Wellcome Trust/United Kingdom ; R21 HD086833/HD/NICHD NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; S10 OD021630/OD/NIH HHS/United States ; R21 AI133522/AI/NIAID NIH HHS/United States ; P30 EY008126/EY/NEI NIH HHS/United States ; P30 CA068485/CA/NCI NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; U24 DK059637/DK/NIDDK NIH HHS/United States ; R01 AI139154/AI/NIAID NIH HHS/United States ; 281668/ERC_/European Research Council/International ; P30 DK020593/DK/NIDDK NIH HHS/United States ; R00 GM114714/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified ; DNA-Binding Proteins/genetics/physiology ; Drosophila/embryology/microbiology/virology ; Drosophila melanogaster/embryology/microbiology/virology ; Female ; Genes, Lethal ; Genes, Viral ; Host Microbial Interactions/genetics/physiology ; Male ; Prophages/*genetics/*pathogenicity/physiology ; Sex Ratio ; Symbiosis/genetics/physiology ; Viral Proteins/genetics/physiology ; Wolbachia/*pathogenicity/*virology ; }, abstract = {Wolbachia are the most widespread maternally-transmitted bacteria in the animal kingdom. Their global spread in arthropods and varied impacts on animal physiology, evolution, and vector control are in part due to parasitic drive systems that enhance the fitness of infected females, the transmitting sex of Wolbachia. Male killing is one common drive mechanism wherein the sons of infected females are selectively killed. Despite decades of research, the gene(s) underlying Wolbachia-induced male killing remain unknown. Here using comparative genomic, transgenic, and cytological approaches in fruit flies, we identify a candidate gene in the eukaryotic association module of Wolbachia prophage WO, termed WO-mediated killing (wmk), which transgenically causes male-specific lethality during early embryogenesis and cytological defects typical of the pathology of male killing. The discovery of wmk establishes new hypotheses for the potential role of phage genes in sex-specific lethality, including the control of arthropod pests and vectors.}, } @article {pmid31500667, year = {2019}, author = {Fisher, ML and Levine, JF and Guy, JS and Mochizuki, H and Breen, M and Schal, C and Watson, DW}, title = {Lack of influence by endosymbiont Wolbachia on virus titer in the common bed bug, Cimex lectularius.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {436}, pmid = {31500667}, issn = {1756-3305}, support = {2016scholarship//W. M. Keck Foundation/ ; }, mesh = {Animals ; Bedbugs/*microbiology/*virology ; Calicivirus, Feline/*growth & development/*isolation & purification ; *Microbial Interactions ; *Viral Load ; Wolbachia/*growth & development ; }, abstract = {BACKGROUND: The common bed bug, Cimex lectularius, is an obligatory blood-feeding ectoparasite that requires a blood meal to molt and produce eggs. Their frequent biting to obtain blood meals and intimate association with humans increase the potential for disease transmission. However, despite more than 100 years of inquiry into bed bugs as potential disease vectors, they still have not been conclusively linked to any pathogen or disease. This ecological niche is extraordinarily rare, given that nearly every other blood-feeding arthropod is associated with some type of human or zoonotic disease. Bed bugs rely on the bacteria Wolbachia as an obligate endosymbiont to biosynthesize B vitamins, since they acquire a nutritionally deficient diet, but it is unknown if Wolbachia confers additional benefits to its bed bug host. In some insects, Wolbachia induces resistance to viruses such as Dengue, Chikungunya, West Nile, Drosophila C and Zika, and primes the insect immune system in other blood-feeding insects. Wolbachia might have evolved a similar role in its mutualistic association with the bed bug. In this study, we evaluated the influence of Wolbachia on virus replication within C. lectularius.

METHODS: We used feline calicivirus as a model pathogen. We fed 40 bed bugs from an established line of Wolbachia-cured and a line of Wolbachia-positive C. lectularius a virus-laden blood meal, and quantified the amount of virus over five time intervals post-feeding. The antibiotic rifampicin was used to cure bed bugs of Wolbachia.

RESULTS: There was a significant effect of time post-feeding, as the amount of virus declined by ~90% over 10 days in both groups, but no significant difference in virus titer was observed between the Wolbachia-positive and Wolbachia-cured groups.

CONCLUSIONS: These findings suggest that other mechanisms are involved in virus suppression within bed bugs, independent of the influence of Wolbachia, and our conclusions underscore the need for future research.}, } @article {pmid31499724, year = {2019}, author = {Zheng, B and Chen, LH and Sun, QW}, title = {Analyzing the control of dengue by releasing Wolbachia-infected male mosquitoes through a delay differential equation model.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {16}, number = {5}, pages = {5531-5550}, doi = {10.3934/mbe.2019275}, pmid = {31499724}, issn = {1551-0018}, mesh = {Aedes/*microbiology/*virology ; Algorithms ; Animals ; Bacterial Infections ; Communicable Disease Control ; Cytoplasm/metabolism ; Dengue/*epidemiology/*prevention & control ; Dengue Virus ; Female ; Humans ; Male ; Models, Theoretical ; Mosquito Vectors/microbiology/virology ; Pest Control, Biological ; *Wolbachia ; }, abstract = {To date, an innovative strategy to control dengue is to release Wolbachia-infected male mosquitoes into wild areas to sterilize wild female mosquito vectors by cytoplasmic incompatibility (CI). To investigate the efficacy of Wolbachia in blocking dengue virus transmission, we develop a deterministic mathematical model of human and mosquito populations in which one dengue serotype circulates. The delay differential equation model captures the respective extrinsic and intrinsic incu-bation periods (EIP and IIP) in the mosquito and human, as well as the maturation delay between mating and emergence of adult mosquitoes, which have received relatively little attention. We analyze the existence and stability of disease-free equilibria, and obtain a sufficient and necessary condition on the existence of the disease-endemic equilibrium. We also determine two threshold values of the release ratio $\theta$, denoted by $\theta_1^*$ and $\theta_2^*$ with $\theta_1^*>\theta_2^*$. When $\theta>\theta_1^*$, the mosquito population will be eradicated eventually. When $\theta_2^*<\theta < \theta_1^*$, a complete mosquito eradication becomes impossible, but virus eradication is ensured at the meantime. When $\theta<\theta_2^*$, the disease-endemic equilibrium emerges that allows dengue virus to circulate between humans and mosquitoes. We carry out sensitivity analysis of the threshold values in terms of the model parameters, and simulate several possible control strate-gies with different release ratios, which confirm the public awareness that reducing mosquito bites and killing adult mosquitoes are the most effective strategy to control the epidemic. Our model provides new insights on the effectiveness of Wolbachia in reducing dengue at a population level.}, } @article {pmid31499687, year = {2019}, author = {Huang, MG and Tang, MX and Yu, JS and Zheng, B}, title = {The impact of mating competitiveness and incomplete cytoplasmic incompatibility on Wolbachia-driven mosquito population suppressio.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {16}, number = {5}, pages = {4741-4757}, doi = {10.3934/mbe.2019238}, pmid = {31499687}, issn = {1551-0018}, mesh = {Aedes/microbiology/physiology/virology ; Animals ; Bioengineering ; China/epidemiology ; Cytoplasm/microbiology ; Dengue/epidemiology/prevention & control/transmission ; Female ; Humans ; Male ; Mathematical Concepts ; Models, Biological ; Mosquito Control/*methods/statistics & numerical data ; Mosquito Vectors/*microbiology/*pathogenicity/virology ; Population Dynamics/statistics & numerical data ; Reproduction ; Wolbachia/*physiology ; }, abstract = {To control mosquito-borne diseases such as dengue, malaria, and Zika, {\it Wolbachia} -infected male mosquitoes have been released in open areas to suppress wild mosquito population driven by cytoplasmic incompatibility (CI). In this work, we initiate a preliminary assessment on how the CI intensity $\xi$, and the mating competitiveness $\mu$ of released males relative to wild males, impact the suppression efficacy by a delay differential equation model. Our analysis identifies a threshold CI intensity $\xi_0\in (0, 1)$ as an increasing function of the natural reproduction rate of the wild mosquitoes, and a threshold value $r^*$ for the ratio $r(t)$ between the numbers of released males and wild males. The population suppression fails when $\xi\le \xi_0$, and succeeds when $\xi>\xi_0$ and $r(t)\ge r^*$. Our analyses indicate that $\xi$ plays a more important role than $\mu$ in the population suppression. For instance, a slight decrease of $\xi$ from 1 to 0.92 is more devastating than halving $\mu$ from 1 to 0.5. In our estimation of the optimal starting date for infected male release to target a more than $95\%$ wild population reduction during the peak season of dengue in Guangzhou, we find that the optimal date is almost independent of $\mu$ but is sensitive to $\xi$. If CI is complete, then starting about two months ahead can be an optimal option for less financial and labor costs. A slight reduction in the CI intensity requires a considerably earlier starting date.}, } @article {pmid31499668, year = {2019}, author = {Liu, YF and Sun, GW and Wang, L and Guo, ZM}, title = {Establishing Wolbachia in the wild mosquito population: The effects of wind and critical patch size.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {16}, number = {5}, pages = {4399-4414}, doi = {10.3934/mbe.2019219}, pmid = {31499668}, issn = {1551-0018}, mesh = {Aedes/*microbiology ; Animals ; Computer Simulation ; Female ; Humans ; Male ; Mathematical Concepts ; Models, Biological ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods/statistics & numerical data ; Population Dynamics/statistics & numerical data ; Vector Borne Diseases/prevention & control/transmission ; Wind ; Wolbachia/*physiology ; }, abstract = {Releasing mosquitoes with Wolbachia into the wild mosquito population is becoming the very promising strategy to control mosquito-borne infections. To investigate the effects of wind and critical patch size on the Wolbachia establishment in the wild mosquito population, in this paper, we propose a diffusion-reaction-advection system in a heterogeneous environment. By studying the related eigenvalue problems, we derive various conditions under which Wolbachia can fully establish in the entire wild mosquito population. Our findings may provide some useful insights on designing practical releasing strategies to control the mosquito population.}, } @article {pmid31499610, year = {2019}, author = {Wang, LX and Shi, PJ and Zhang, HG}, title = {Bifurcation analysis of a wild and sterile mosquito model.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {16}, number = {5}, pages = {3215-3234}, doi = {10.3934/mbe.2019160}, pmid = {31499610}, issn = {1551-0018}, mesh = {Algorithms ; Animals ; California ; Computer Simulation ; Culicidae/microbiology/*physiology ; Female ; *Infertility ; Male ; Models, Biological ; Mosquito Vectors ; Pest Control, Biological/*methods/statistics & numerical data ; Population Dynamics ; Wolbachia ; }, abstract = {The bifurcation of an ordinary differential equation model describing interaction of the wild and the released sterile mosquitoes is analyzed. It is shown that the model undergoes a sequence of bifurcations including saddle-node bifurcation, supercritical Hopf bifurcation, subcritical Hopf bifurcation, homoclinic bifurcation and Bogdanov-Takens bifurcation. We also find that the model displays monostable, bistable or tristable dynamics. This analysis suggests that the densities of the initial wild mosquitoes and the released sterile ones determine the asymptotic states of both populations. This study may give an insight into the estimation number of the released sterile mosquitoes.}, } @article {pmid31497407, year = {2019}, author = {Ning, SF and Zhou, JC and Liu, QQ and Zhao, Q and Dong, H}, title = {Gradual, temperature-induced change of secondary sexual characteristics in Trichogramma pretiosum infected with parthenogenesis-inducing Wolbachia.}, journal = {PeerJ}, volume = {7}, number = {}, pages = {e7567}, pmid = {31497407}, issn = {2167-8359}, abstract = {Intersex is an intermediate stage of sexual differentiation in insects. Determining intersex morphology and the cause of its production will aid in the understanding of the mechanism of sexual differentiation in insects. In this paper, Wolbachia-infected Trichogramma pretiosum (T. preW[+]) that shows thelytokous parthenogenesis were used as subjects. In order to determine the causes of the T. preW[+] intersex and the influence of parental generation temperature on gradual changes in secondary masculinization in intersex offspring, we examined the occurrence of intersex offspring (F1 and F2 generation) after the parental generations were treated with high temperature (27, 29, 31, and 33 °C) and described the external morphology of the intersexes. The results showed that the T. preW[+] parental generation temperature is positively correlated with the probability of intersex offspring. The probability of F1 intersex is significantly higher than that of F2 intersex in different high temperature. The degree of secondary masculinization in T. preW[+] intersexes increases as parental generation temperature increases. In addition, our study first identified 11 intersex types in T. preW[+] and found that the primary and secondary sexual characteristics showed a regular distribution. We also found that the D type and H type of intersex have the highest frequency of appearance. The external genitalia of most intersexes were female, and only three intersex types have male external genitalia. Conclusions were ultimately obtained: Wolbachia is a direct factor that causes the occurrence of intersexes, while high temperature is an indirect factor that determines the external morphology of intersexes. The effects of high temperature on T. preW[+] intersexes is passed through the parental generation to offspring, and this maternal effect weakens as the number of generations increases. In T. preW[+] intersex individuals, most exhibit female primary sexual characteristics, and secondary sexual characteristics exhibit signs of masculinization.}, } @article {pmid31496523, year = {2019}, author = {Wan Sulaiman, WA and Kamtchum-Tatuene, J and Mohamed, MH and Ramachandran, V and Ching, SM and Sazlly Lim, SM and Hashim, HZ and Inche Mat, LN and Hoo, FK and Basri, H}, title = {Anti-Wolbachia therapy for onchocerciasis & lymphatic filariasis: Current perspectives.}, journal = {The Indian journal of medical research}, volume = {149}, number = {6}, pages = {706-714}, pmid = {31496523}, issn = {0971-5916}, mesh = {Adult ; Albendazole/therapeutic use ; Animals ; Anti-Bacterial Agents/therapeutic use ; Doxycycline/therapeutic use ; Drug Therapy, Combination ; Elephantiasis, Filarial/*drug therapy/epidemiology/microbiology ; Humans ; India/epidemiology ; Neglected Diseases/*drug therapy/epidemiology/microbiology ; Onchocerciasis/*drug therapy/epidemiology/microbiology ; Tropical Medicine ; Wolbachia/drug effects/*pathogenicity ; }, abstract = {Onchocerciasis and lymphatic filariasis (LF) are human filarial diseases belonging to the group of neglected tropical diseases, leading to permanent and long-term disability in infected individuals in the endemic countries such as Africa and India. Microfilaricidal drugs such as ivermectin and albendazole have been used as the standard therapy in filariasis, although their efficacy in eliminating the diseases is not fully established. Anti-Wolbachia therapy employs antibiotics and is a promising approach showing potent macrofilaricidal activity and also prevents embryogenesis. This has translated to clinical benefits resulting in successful eradication of microfilarial burden, thus averting the risk of adverse events from target species as well as those due to co-infection with loiasis. Doxycycline shows potential as an anti-Wolbachia treatment, leading to the death of adult parasitic worms. It is readily available, cheap and safe to use in adult non-pregnant patients. Besides doxycycline, several other potential antibiotics are also being investigated for the treatment of LF and onchocerciasis. This review aims to discuss and summarise recent developments in the use of anti-Wolbachia drugs to treat onchocerciasis and LF.}, } @article {pmid31495336, year = {2019}, author = {O'Reilly, KM and Hendrickx, E and Kharisma, DD and Wilastonegoro, NN and Carrington, LB and Elyazar, IRF and Kucharski, AJ and Lowe, R and Flasche, S and Pigott, DM and Reiner, RC and Edmunds, WJ and Hay, SI and Yakob, L and Shepard, DS and Brady, OJ}, title = {Estimating the burden of dengue and the impact of release of wMel Wolbachia-infected mosquitoes in Indonesia: a modelling study.}, journal = {BMC medicine}, volume = {17}, number = {1}, pages = {172}, pmid = {31495336}, issn = {1741-7015}, support = {206471/Z/17/Z/WT_/Wellcome Trust/United Kingdom ; 206250/Z/17/Z/WT_/Wellcome Trust/United Kingdom ; 208812/Z/17/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; Cost of Illness ; Dengue/epidemiology/*prevention & control/transmission ; Dengue Virus ; Humans ; Indonesia/epidemiology ; *Models, Theoretical ; Pest Control, Biological/*methods ; *Wolbachia ; }, abstract = {BACKGROUND: Wolbachia-infected mosquitoes reduce dengue virus transmission, and city-wide releases in Yogyakarta city, Indonesia, are showing promising entomological results. Accurate estimates of the burden of dengue, its spatial distribution and the potential impact of Wolbachia are critical in guiding funder and government decisions on its future wider use.

METHODS: Here, we combine multiple modelling methods for burden estimation to predict national case burden disaggregated by severity and map the distribution of burden across the country using three separate data sources. An ensemble of transmission models then predicts the estimated reduction in dengue transmission following a nationwide roll-out of wMel Wolbachia.

RESULTS: We estimate that 7.8 million (95% uncertainty interval [UI] 1.8-17.7 million) symptomatic dengue cases occurred in Indonesia in 2015 and were associated with 332,865 (UI 94,175-754,203) lost disability-adjusted life years (DALYs). The majority of dengue's burden was due to non-severe cases that did not seek treatment or were challenging to diagnose in outpatient settings leading to substantial underreporting. Estimated burden was highly concentrated in a small number of large cities with 90% of dengue cases occurring in 15.3% of land area. Implementing a nationwide Wolbachia population replacement programme was estimated to avert 86.2% (UI 36.2-99.9%) of cases over a long-term average.

CONCLUSIONS: These results suggest interventions targeted to the highest burden cities can have a disproportionate impact on dengue burden. Area-wide interventions, such as Wolbachia, that are deployed based on the area covered could protect people more efficiently than individual-based interventions, such as vaccines, in such dense environments.}, } @article {pmid31491378, year = {2019}, author = {Zheng, Y and Shen, W and Bi, J and Chen, MY and Wang, RF and Ai, H and Wang, YF}, title = {Small RNA analysis provides new insights into cytoplasmic incompatibility in Drosophila melanogaster induced by Wolbachia.}, journal = {Journal of insect physiology}, volume = {118}, number = {}, pages = {103938}, doi = {10.1016/j.jinsphys.2019.103938}, pmid = {31491378}, issn = {1879-1611}, mesh = {3' Untranslated Regions ; Animals ; Cytoplasm/*genetics/pathology ; Drosophila melanogaster/embryology/genetics/metabolism/*microbiology ; Female ; *Gene Expression Regulation, Developmental ; Male ; MicroRNAs/genetics ; RNA, Small Untranslated ; Symbiosis ; Testis ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a genus of endosymbiotic bacteria that induce a wide range of effects on their insect hosts. Cytoplasmic incompatibility (CI) is the most common phenotype mediated by Wolbachia and results in embryonic lethality when Wolbachia-infected males mate with uninfected females. Studies have revealed that bacteria can regulate many cellular processes in their hosts using small non-coding RNAs, so we investigated the involvement of small RNAs (sRNAs) in CI. Comparison of sRNA libraries between Wolbachia-infected and uninfected Drosophila melanogaster testes revealed 18 novel microRNAs (miRNAs), of which 12 were expressed specifically in Wolbachia-infected flies and one specifically in Wolbachia-uninfected flies. Furthermore, ten miRNAs showed differential expression, with four upregulated and six downregulated in Wolbachia-infected flies. Of the upregulated miRNAs, nov-miR-12 exhibited the highest upregulation in the testes of D. melanogaster. We then identified pipsqueak (psq) as the target gene of nov-miR-12 with the greatest complementarity in its 3' untranslated region (UTR). Wolbachia infection was correlated with reduced psq expression in D. melanogaster, and luciferase assays demonstrated that nov-miR-12 could downregulate psq through binding to its 3'UTR region. Knockdown of psq in Wolbachia-free fly testes significantly reduced egg hatching rate and mimicked the cellular abnormalities of Wolbachia-induced CI in embryos, including asynchronous nuclear division, chromatin bridging, and chromatin fragmentation. These results suggest that Wolbachia may induce CI in insect hosts by miRNA-mediated changes in host gene expression. Moreover, these findings reveal a potential molecular strategy for elucidating the complex interactions between endosymbionts and their insect hosts, such as Wolbachia-driven CI.}, } @article {pmid31488182, year = {2019}, author = {Rohde, BB and Staunton, KM and Zeak, NC and Beebe, N and Snoad, N and Bondarenco, A and Liddington, C and Anderson, JA and Xiang, W and Mankin, RW and Ritchie, SA}, title = {Waterproof, low-cost, long-battery-life sound trap for surveillance of male Aedes aegypti for rear-and-release mosquito control programmes.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {417}, pmid = {31488182}, issn = {1756-3305}, support = {DGE 1315138//National Science Foundation Graduate Research Fellowship/ ; }, mesh = {Aedes/*physiology/virology ; Animals ; Costs and Cost Analysis ; Dengue/prevention & control ; Female ; Male ; Mosquito Control/economics/*instrumentation/*methods ; Mosquito Vectors/physiology/virology ; *Sound ; Virus Diseases/*prevention & control ; Zika Virus Infection/prevention & control/transmission ; }, abstract = {BACKGROUND: Sterile male rear-and-release programmes are of growing interest for controlling Aedes aegypti, including use an "incompatible insect technique" (IIT) to suppress transmission of dengue, Zika, and other viruses. Under IIT, males infected with Wolbachia are released into the suppression area to induce cytoplasmic incompatibility in uninfected populations. These and similar mosquito-release programmes require cost-effective field surveys of both sexes to optimize the locations, timing, and quantity of releases. Unfortunately, traps that sample male Ae. aegypti effectively are expensive and usually require mains power. Recently, an electronic lure was developed that attracts males using a 484 Hz sinusoidal tone mimicking the female wingbeat frequencies, broadcast in a 120 s on/off cycle. When deployed in commercially available gravid Aedes traps (GATs), the new combination, sound-GAT (SGAT), captures both males and females effectively. Given its success, there is interest in optimizing SGAT to reduce cost and power usage while maximizing catch rates.

METHODS: Options considered in this study included use of a smaller, lower-power microcontroller (Tiny) with either the original or a lower-cost speaker (lcS). A 30 s on/off cycle was tested in addition to the original 120 s cycle to minimize the potential that the longer cycle induced habituation. The original SGAT was compared against other traps incorporating the Tiny-based lures for mosquito capture in a large semi-field cage. The catch rates in waterproofed versions of this trap were then compared with catch rates in standard [BG-Sentinel 2 (BGS 2); Biogents AG, Regensburg, Germany] traps during an IIT field study in the Innisfail region of Queensland, Australia in 2017.

RESULTS: The system with a low-power microcontroller and low-cost speaker playing a 30 s tone (Tiny-lcS-30s) caught the highest proportion of males. The mean proportions of males caught in a semi-field cage were not significantly different among the original design and the four low-power, low-cost versions of the SGAT. During the IIT field study, the waterproofed version of the highest-rated, Tiny-lcS-30s SGAT captured male Ae. aegypti at similar rates as co-located BGS-2 traps.

CONCLUSIONS: Power- and cost-optimized, waterproofed versions of male Ae. aegypti acoustic lures in GATs are now available for field use in areas with sterile male mosquito rear-and-release programmes.}, } @article {pmid31481018, year = {2019}, author = {Christensen, S and Camacho, M and Sharmin, Z and Momtaz, AJMZ and Perez, L and Navarro, G and Triana, J and Samarah, H and Turelli, M and Serbus, LR}, title = {Quantitative methods for assessing local and bodywide contributions to Wolbachia titer in maternal germline cells of Drosophila.}, journal = {BMC microbiology}, volume = {19}, number = {1}, pages = {206}, pmid = {31481018}, issn = {1471-2180}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animal Feed/analysis ; Animals ; Cytological Techniques/*methods ; Drosophila melanogaster/metabolism/*microbiology ; Female ; Ovary/growth & development/microbiology ; Ovum/growth & development/*microbiology ; Polymerase Chain Reaction/*methods ; Wolbachia/genetics/*growth & development/isolation & purification ; }, abstract = {BACKGROUND: Little is known about how bacterial endosymbionts colonize host tissues. Because many insect endosymbionts are maternally transmitted, egg colonization is critical for endosymbiont success. Wolbachia bacteria, carried by approximately half of all insect species, provide an excellent model for characterizing endosymbiont infection dynamics. To date, technical limitations have precluded stepwise analysis of germline colonization by Wolbachia. It is not clear to what extent titer-altering effects are primarily mediated by growth rates of Wolbachia within cell lineages or migration of Wolbachia between cells.

RESULTS: The objective of this work is to inform mechanisms of germline colonization through use of optimized methodology. The approaches are framed in terms of nutritional impacts on Wolbachia. Yeast-rich diets in particular have been shown to suppress Wolbachia titer in the Drosophila melanogaster germline. To determine the extent of Wolbachia sensitivity to diet, we optimized 3-dimensional, multi-stage quantification of Wolbachia titer in maternal germline cells. Technical and statistical validation confirmed the identity of Wolbachia in vivo, the reproducibility of Wolbachia quantification and the statistical power to detect these effects. The data from adult feeding experiments demonstrated that germline Wolbachia titer is distinctly sensitive to yeast-rich host diets in late oogenesis. To investigate the physiological basis for these nutritional impacts, we optimized methodology for absolute Wolbachia quantification by real-time qPCR. We found that yeast-rich diets exerted no significant effect on bodywide Wolbachia titer, although ovarian titers were significantly reduced. This suggests that host diets affects Wolbachia distribution between the soma and late stage germline cells. Notably, relative qPCR methods distorted apparent wsp abundance, due to altered host DNA copy number in yeast-rich conditions. This highlights the importance of absolute quantification data for testing mechanistic hypotheses.

CONCLUSIONS: We demonstrate that absolute quantification of Wolbachia, using well-controlled cytological and qPCR-based methods, creates new opportunities to determine how bacterial abundance within the germline relates to bacterial distribution within the body. This methodology can be applied to further test germline infection dynamics in response to chemical treatments, genetic conditions, new host/endosymbiont combinations, or potentially adapted to analyze other cell and tissue types.}, } @article {pmid31477173, year = {2019}, author = {Attardo, GM and Abd-Alla, AMM and Acosta-Serrano, A and Allen, JE and Bateta, R and Benoit, JB and Bourtzis, K and Caers, J and Caljon, G and Christensen, MB and Farrow, DW and Friedrich, M and Hua-Van, A and Jennings, EC and Larkin, DM and Lawson, D and Lehane, MJ and Lenis, VP and Lowy-Gallego, E and Macharia, RW and Malacrida, AR and Marco, HG and Masiga, D and Maslen, GL and Matetovici, I and Meisel, RP and Meki, I and Michalkova, V and Miller, WJ and Minx, P and Mireji, PO and Ometto, L and Parker, AG and Rio, R and Rose, C and Rosendale, AJ and Rota-Stabelli, O and Savini, G and Schoofs, L and Scolari, F and Swain, MT and Takáč, P and Tomlinson, C and Tsiamis, G and Van Den Abbeele, J and Vigneron, A and Wang, J and Warren, WC and Waterhouse, RM and Weirauch, MT and Weiss, BL and Wilson, RK and Zhao, X and Aksoy, S}, title = {Comparative genomic analysis of six Glossina genomes, vectors of African trypanosomes.}, journal = {Genome biology}, volume = {20}, number = {1}, pages = {187}, pmid = {31477173}, issn = {1474-760X}, support = {D43 TW007391/TW/FIC NIH HHS/United States ; R01AI051584/NH/NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; R03TW008413/NH/NIH HHS/United States ; U54HG003079/HG/NHGRI NIH HHS/United States ; U01 AI115648/AI/NIAID NIH HHS/United States ; R03TW009444/NH/NIH HHS/United States ; U01AI115648/NH/NIH HHS/United States ; R21AI109263//National Institute of Allergy and Infectious Diseases/International ; }, mesh = {Animals ; DNA Transposable Elements/genetics ; Drosophila melanogaster/genetics ; Female ; Gene Expression Regulation ; Genes, Insect ; Genes, X-Linked ; *Genome, Insect ; *Genomics ; Geography ; Insect Proteins/genetics ; Insect Vectors/*genetics ; Male ; Mutagenesis, Insertional/genetics ; Phylogeny ; Repetitive Sequences, Nucleic Acid/genetics ; Sequence Homology, Amino Acid ; Synteny/genetics ; Trypanosoma/*parasitology ; Tsetse Flies/*genetics ; Wolbachia/genetics ; }, abstract = {BACKGROUND: Tsetse flies (Glossina sp.) are the vectors of human and animal trypanosomiasis throughout sub-Saharan Africa. Tsetse flies are distinguished from other Diptera by unique adaptations, including lactation and the birthing of live young (obligate viviparity), a vertebrate blood-specific diet by both sexes, and obligate bacterial symbiosis. This work describes the comparative analysis of six Glossina genomes representing three sub-genera: Morsitans (G. morsitans morsitans, G. pallidipes, G. austeni), Palpalis (G. palpalis, G. fuscipes), and Fusca (G. brevipalpis) which represent different habitats, host preferences, and vectorial capacity.

RESULTS: Genomic analyses validate established evolutionary relationships and sub-genera. Syntenic analysis of Glossina relative to Drosophila melanogaster shows reduced structural conservation across the sex-linked X chromosome. Sex-linked scaffolds show increased rates of female-specific gene expression and lower evolutionary rates relative to autosome associated genes. Tsetse-specific genes are enriched in protease, odorant-binding, and helicase activities. Lactation-associated genes are conserved across all Glossina species while male seminal proteins are rapidly evolving. Olfactory and gustatory genes are reduced across the genus relative to other insects. Vision-associated Rhodopsin genes show conservation of motion detection/tracking functions and variance in the Rhodopsin detecting colors in the blue wavelength ranges.

CONCLUSIONS: Expanded genomic discoveries reveal the genetics underlying Glossina biology and provide a rich body of knowledge for basic science and disease control. They also provide insight into the evolutionary biology underlying novel adaptations and are relevant to applied aspects of vector control such as trap design and discovery of novel pest and disease control strategies.}, } @article {pmid31471686, year = {2019}, author = {Li, Y and Liu, X and Guo, H}, title = {Population Dynamics of Wolbachia in Laodelphax striatellus (Fallén) Under Successive Stress of Antibiotics.}, journal = {Current microbiology}, volume = {76}, number = {11}, pages = {1306-1312}, pmid = {31471686}, issn = {1432-0991}, support = {31672027//the National Natural Science Foundation of China/ ; cx(16)1001//the Independent Innovation Fund of Agricultural Science and Technology in Jiangsu province, China/ ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Biodiversity ; Female ; Hemiptera/*microbiology ; Male ; Tetracycline/*pharmacology ; Wolbachia/*drug effects/growth & development ; }, abstract = {Wolbachia are the most common symbionts in arthropods; antibiotic treatment for eliminating the symbionts from their host is necessary to investigate the functions. Tetracycline antibiotics are widely used to remove endosymbiont Wolbachia from insect hosts. However, very little has been known on the effects of tetracycline on population size of Wolbachia in small brown planthopper (SBPH), Laodelphax striatellus (Fallén), an important insect pest of rice in Asia. Here, we investigated the dynamics of Wolbachia population density in females and males of L. striatellus by real-time fluorescent quantitative PCR method. The Wolbachia density in females and males of L. striatellus all declined sharply after treatment with 2 mg/mL tetracycline for one generation, and continued to decrease to a level which could not be detected by both qPCR and diagnostic PCR after treated for another generation, then maintained at 0 in the following three generations with continuous antibiotic treatment. Wolbachia infection did not recover in L. striatellus after stopping tetracycline treatment for ten generations. This is the first report to precisely monitor the population dynamics of Wolbachia in L. striatellus during successive tetracycline treatment and after that. The results provide a useful method for evaluating the efficiency of artificial operation of endosymbionts.}, } @article {pmid31469838, year = {2019}, author = {Jiménez, NE and Gerdtzen, ZP and Olivera-Nappa, Á and Salgado, JC and Conca, C}, title = {A systems biology approach for studying Wolbachia metabolism reveals points of interaction with its host in the context of arboviral infection.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {8}, pages = {e0007678}, pmid = {31469838}, issn = {1935-2735}, mesh = {Aedes/microbiology ; Animals ; Drosophila melanogaster/microbiology ; *Host Microbial Interactions ; Hymenoptera/microbiology ; *Symbiosis ; Systems Biology/methods ; Wolbachia/*growth & development/*metabolism ; }, abstract = {Wolbachia are alpha-proteobacteria known to infect arthropods, which are of interest for disease control since they have been associated with improved resistance to viral infection. Although several genomes for different strains have been sequenced, there is little knowledge regarding the relationship between this bacterium and their hosts, particularly on their dependency for survival. Motivated by the potential applications on disease control, we developed genome-scale models of four Wolbachia strains known to infect arthropods: wAlbB (Aedes albopictus), wVitA (Nasonia vitripennis), wMel and wMelPop (Drosophila melanogaster). The obtained metabolic reconstructions exhibit a metabolism relying mainly on amino acids for energy production and biomass synthesis. A gap analysis was performed to detect metabolic candidates which could explain the endosymbiotic nature of this bacterium, finding that amino acids, requirements for ubiquinone precursors and provisioning of metabolites such as riboflavin could play a crucial role in this relationship. This work provides a systems biology perspective for studying the relationship of Wolbachia with its host and the development of new approaches for control of the spread of arboviral diseases. This approach, where metabolic gaps are key objects of study instead of just additions to complete a model, could be applied to other endosymbiotic bacteria of interest.}, } @article {pmid31462916, year = {2019}, author = {Ayala, D and Akone-Ella, O and Rahola, N and Kengne, P and Ngangue, MF and Mezeme, F and Makanga, BK and Nigg, M and Costantini, C and Simard, F and Prugnolle, F and Roche, B and Duron, O and Paupy, C}, title = {Natural Wolbachia infections are common in the major malaria vectors in Central Africa.}, journal = {Evolutionary applications}, volume = {12}, number = {8}, pages = {1583-1594}, pmid = {31462916}, issn = {1752-4571}, abstract = {During the last decade, the endosymbiont bacterium Wolbachia has emerged as a biological tool for vector disease control. However, for long time, it was believed that Wolbachia was absent in natural populations of Anopheles. The recent discovery that species within the Anopheles gambiae complex host Wolbachia in natural conditions has opened new opportunities for malaria control research in Africa. Here, we investigated the prevalence and diversity of Wolbachia infection in 25 African Anopheles species in Gabon (Central Africa). Our results revealed the presence of Wolbachia in 16 of these species, including the major malaria vectors in this area. The infection prevalence varied greatly among species, confirming that sample size is a key factor to detect the infection. Moreover, our sequencing and phylogenetic analyses showed the important diversity of Wolbachia strains that infect Anopheles. Co-evolutionary analysis unveiled patterns of Wolbachia transmission within some Anopheles species, suggesting that past independent acquisition events were followed by co-cladogenesis. The large diversity of Wolbachia strains that infect natural populations of Anopheles offers a promising opportunity to select suitable phenotypes for suppressing Plasmodium transmission and/or manipulating Anopheles reproduction, which in turn could be used to reduce the malaria burden in Africa.}, } @article {pmid31462331, year = {2019}, author = {Shi, C and Beller, L and Deboutte, W and Yinda, KC and Delang, L and Vega-Rúa, A and Failloux, AB and Matthijnssens, J}, title = {Stable distinct core eukaryotic viromes in different mosquito species from Guadeloupe, using single mosquito viral metagenomics.}, journal = {Microbiome}, volume = {7}, number = {1}, pages = {121}, pmid = {31462331}, issn = {2049-2618}, mesh = {Aedes/*virology ; Animals ; Culex/*virology ; Genome, Viral/*genetics ; Guadeloupe ; Metagenome/*genetics ; Metagenomics ; Mosquito Vectors/*virology ; Phylogeny ; *Viruses/classification/genetics/isolation & purification ; }, abstract = {BACKGROUND: Mosquitoes are the most important invertebrate viral vectors in humans and harbor a high diversity of understudied viruses, which has been shown in many mosquito virome studies in recent years. These studies generally performed metagenomics sequencing on pools of mosquitoes, without assessment of the viral diversity in individual mosquitoes. To address this issue, we applied our optimized viral metagenomics protocol (NetoVIR) to compare the virome of single and pooled Aedes aegypti and Culex quinquefasciatus mosquitoes collected from different locations in Guadeloupe, in 2016 and 2017.

RESULTS: The total read number and viral reads proportion of samples containing a single mosquito have no significant difference compared with those of pools containing five mosquitoes, which proved the feasibility of using single mosquito for viral metagenomics. A comparative analysis of the virome revealed a higher abundance and more diverse eukaryotic virome in Aedes aegypti, whereas Culex quinquefasciatus harbors a richer and more diverse phageome. The majority of the identified eukaryotic viruses were mosquito-species specific. We further characterized the genomes of 11 novel eukaryotic viruses. Furthermore, qRT-PCR analyses of the six most abundant eukaryotic viruses indicated that the majority of individual mosquitoes were infected by several of the selected viruses with viral genome copies per mosquito ranging from 267 to 1.01 × 10[8] (median 7.5 × 10[6]) for Ae. aegypti and 192 to 8.69 × 10[6] (median 4.87 × 10[4]) for Cx. quinquefasciatus. Additionally, in Cx. quinquefasciatus, a number of phage contigs co-occurred with several marker genes of Wolbachia sp. strain wPip.

CONCLUSIONS: We firstly demonstrate the feasibility to use single mosquito for viral metagenomics, which can provide much more precise virome profiles of mosquito populations. Interspecific comparisons show striking differences in abundance and diversity between the viromes of Ae. aegypti and Cx. quinquefasciatus. Those two mosquito species seem to have their own relatively stable "core eukaryotic virome", which might have important implications for the competence to transmit important medically relevant arboviruses. The presence of Wolbachia in Cx. quinquefasciatus might explain (1) the lower overall viral load compared to Ae. aegypti, (2) the identification of multiple unknown phage contigs, and (3) the difference in competence for important human pathogens. How these viruses, phages, and bacteria influence the physiology and vector competence of mosquito hosts warrants further research.}, } @article {pmid31451771, year = {2019}, author = {Ford, SA and Allen, SL and Ohm, JR and Sigle, LT and Sebastian, A and Albert, I and Chenoweth, SF and McGraw, EA}, title = {Selection on Aedes aegypti alters Wolbachia-mediated dengue virus blocking and fitness.}, journal = {Nature microbiology}, volume = {4}, number = {11}, pages = {1832-1839}, pmid = {31451771}, issn = {2058-5276}, support = {R01 AI143758/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*genetics/microbiology/virology ; Animals ; Dengue Virus/genetics/*pathogenicity ; Evolution, Molecular ; Female ; Gene Frequency ; Genetic Fitness ; Genome, Bacterial ; Insect Proteins/*genetics ; Male ; Mosquito Vectors/physiology ; Polymorphism, Single Nucleotide ; Wolbachia/*physiology ; }, abstract = {The dengue, Zika and chikungunya viruses are transmitted by the mosquito Aedes aegypti and pose a substantial threat to global public health. Current vaccines and mosquito control strategies have limited efficacy, so novel interventions are needed[1,2]. Wolbachia are bacteria that inhabit insect cells and have been found to reduce viral infection-a phenotype that is referred to as viral 'blocking'[3]. Although not naturally found in A. aegypti[4], Wolbachia were stably introduced into this mosquito in 2011[4,5] and were shown to reduce the transmission potential of dengue, Zika and chikungunya[6,7]. Subsequent field trials showed Wolbachia's ability to spread through A. aegypti populations and reduce the local incidence of dengue fever[8]. Despite these successes, the evolutionary stability of viral blocking is unknown. Here, we utilized artificial selection to reveal genetic variation in the mosquito that affects Wolbachia-mediated dengue blocking. We found that mosquitoes exhibiting weaker blocking also have reduced fitness, suggesting the potential for natural selection to maintain blocking. We also identified A. aegypti genes that affect blocking strength, shedding light on a possible mechanism for the trait. These results will inform the use of Wolbachia as biocontrol agents against mosquito-borne viruses and direct further research into measuring and improving their efficacy.}, } @article {pmid31442887, year = {2019}, author = {Genchi, M and Vismarra, A and Lucchetti, C and Viglietti, A and Crosara, S and Gnudi, G and Quintavalla, C and Schaper, R and Kramer, L}, title = {Efficacy of imidacloprid 10%/moxidectin 2.5% spot on (Advocate®, Advantage Multi®) and doxycycline for the treatment of natural Dirofilaria immitis infections in dogs.}, journal = {Veterinary parasitology}, volume = {273}, number = {}, pages = {11-16}, doi = {10.1016/j.vetpar.2019.07.011}, pmid = {31442887}, issn = {1873-2550}, mesh = {Animals ; Antigens, Helminth/blood ; Arsenicals/therapeutic use ; Dirofilaria immitis ; Dirofilariasis/*drug therapy ; Dog Diseases/drug therapy ; Dogs ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; Female ; Filaricides/therapeutic use ; Macrolides/*therapeutic use ; Male ; Neonicotinoids/*therapeutic use ; Nitro Compounds/*therapeutic use ; Time Factors ; Treatment Outcome ; Triazines/therapeutic use ; }, abstract = {Heartworm infection (also known as dirofilariosis due to Dirofilaria immitis) in dogs causes chronic pulmonary disease that, if left untreated, can lead to right-side congestive heart failure. Currently, the only registered drug for adulticide therapy in dogs with heartworm disease (HWD) is melarsomine dihydrochloride. The recent targeting of the bacterial endosymbiont Wolbachia, through antibiotic therapy of the infected host, has offered an interesting alternative for the treatment of HWD. Recent reports of the adulticide activity of an ivermectin/doxycycline combination protocol has lead the American Heartworm Society (AHS) to include in its guidelines that, in cases where arsenical therapy is not possible or is contraindicated, a monthly heartworm preventive along with doxycycline for a 4-week period might be considered. In the present study, 20 dogs with confirmed natural D. immitis infection were included following owner consent. Fourteen dogs were treated with a topical formulation containing 10% w/v imidacloprid and 2.5% w/v moxidectin (Advocate®, Advantage Multi®, Bayer), monthly for nine months, associated to doxycycline (10 mg/kg/BID) for the first 30 days. Six dogs were treated with melarsomine (Immiticide®, Merial) (2.5 mg/kg) at enrollment, followed one month later by two injections 24 h apart. The presence of circulating antigens and the number of microfilariae (mf) were evaluated at the moment of enrollment and then at 1, 2, 3, 4, 5, 6, 7, 8, 12, 18, 24 months post enrollment. Echocardiogram and radiographs were performed at month 0, 6, 12, 18, 24. Monthly moxidectin combined with 30 days of doxycycline eliminated circulating microfilariae within one month, thus breaking the transmission cycle very quickly. Furthermore, dogs treated with the combination protocol started to become negative for circulating antigens at 4 months from the beginning of treatment and all except one were antigen negative at 9 months. All dogs treated with melarsomine were antigen negative by 5 months from the beginning of the treatment. No dogs showed worsening of pulmonary patterns or criteria indicative of pulmonary hypertension 12 to 24 months after. For the criteria mf concentration, antigen concentration, radiography and echocardiography at 12, 18 and 24 months the non-inferiority for the moxidectin group could be proven for a non-inferiority margin of 15% for the rate difference. Dogs treated with moxidectin and doxycycline became negative for microfilariae and antigens sooner when compared to melarsomine in the present study and to dogs treated with doxycycline combined with ivermectin in studies previously published.}, } @article {pmid31442178, year = {2019}, author = {Cohnstaedt, LW and Alfonso-Parra, C}, title = {2018 Highlights of Mosquito and Vector Biology and Control in Latin America.}, journal = {Journal of the American Mosquito Control Association}, volume = {35}, number = {1}, pages = {40-46}, doi = {10.2987/18-6769.1}, pmid = {31442178}, issn = {1943-6270}, mesh = {Animals ; *Culicidae ; Latin America ; *Mosquito Control ; *Mosquito Vectors ; }, abstract = {The 28th Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 84th Annual Meeting of the AMCA held in Kansas City, MO, in February 2018. The Latin American Symposium promotes the participation of vector control specialists, public health workers, and academic members from Latin America and the sharing of scientific data between continents. Generally, presentations are in Spanish with simultaneous translation into English. The majority of presentation slides are in English to facilitate communication among all meeting attendees. This publication includes summaries of 14 oral presentations by participants from Colombia, Mexico, and the United States of America. Topics addressed in the 3 sessions of the symposium included: larval habitats, insecticide resistance, new Aedes mosquito traps, blood meal analysis and fitness outcomes, vertical transmission of dengue, and transstadial composition of midgut microbiota. Control techniques discussed included sterile insect technique (SIT) with radiation, SIT by Wolbachia, thermal fogging, ultra-low volume pesticide applications, indoor residual spraying, Bacillus thuringiensis israelensis de Barjac (Bti), and Spinosad larval treatments. Presentations were also given on species composition and diversity of phlebotomine sand flies. Presentations were related to the vector species belonging to the mosquito genera, Anopheles, Aedes, and Culex as well as phlebotomine sand flies involved in the transmission of the causal agents of malaria, arboviruses (dengue, chikungunya, Zika), and leishmaniasis.}, } @article {pmid31434963, year = {2019}, author = {Bennett, KL and Gómez-Martínez, C and Chin, Y and Saltonstall, K and McMillan, WO and Rovira, JR and Loaiza, JR}, title = {Dynamics and diversity of bacteria associated with the disease vectors Aedes aegypti and Aedes albopictus.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {12160}, pmid = {31434963}, issn = {2045-2322}, mesh = {Aedes/growth & development/*microbiology ; Animals ; Bacteria/genetics/*growth & development ; Disease Vectors ; Ecosystem ; Hydrogen-Ion Concentration ; Larva/microbiology ; *Microbiota ; Panama ; Principal Component Analysis ; RNA, Ribosomal, 16S/chemistry/genetics/metabolism ; Sequence Analysis, DNA ; Temperature ; Water/chemistry ; Wolbachia/genetics/growth & development ; }, abstract = {Aedes aegypti and Aedes albopictus develop in the same aquatic sites where they encounter microorganisms that influence their life history and capacity to transmit human arboviruses. Some bacteria such as Wolbachia are currently being considered for the control of Dengue, Chikungunya and Zika. Yet little is known about the dynamics and diversity of Aedes-associated bacteria, including larval habitat features that shape their tempo-spatial distribution. We applied large-scale 16S rRNA amplicon sequencing to 960 adults and larvae of both Ae. aegypti and Ae. albopictus mosquitoes from 59 sampling sites widely distributed across nine provinces of Panama. We find both species share a limited, yet highly variable core microbiota, reflecting high stochasticity within their oviposition habitats. Despite sharing a large proportion of microbiota, Ae. aegypti harbours higher bacterial diversity than Ae. albopictus, primarily due to rarer bacterial groups at the larval stage. We find significant differences between the bacterial communities of larvae and adult mosquitoes, and among samples from metal and ceramic containers. However, we find little support for geography, water temperature and pH as predictors of bacterial associates. We report a low incidence of natural Wolbachia infection for both Aedes and its geographical distribution. This baseline information provides a foundation for studies on the functions and interactions of Aedes-associated bacteria with consequences for bio-control within Panama.}, } @article {pmid31432763, year = {2019}, author = {Charlesworth, J and Weinert, LA and Araujo, EV and Welch, JJ}, title = {Wolbachia, Cardinium and climate: an analysis of global data.}, journal = {Biology letters}, volume = {15}, number = {8}, pages = {20190273}, pmid = {31432763}, issn = {1744-957X}, support = {109385/Z/15/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Arthropods ; Bacteroidetes ; Insecta ; Symbiosis ; *Wolbachia ; }, abstract = {Bacterial endosymbionts are very common in terrestrial arthropods, but infection levels vary widely among populations. Experiments and within-species comparisons suggest that environmental temperature might be important in explaining this variation. To investigate the importance of temperature, at broad geographical and taxonomic scales, we extended a global database of terrestrial arthropods screened for Wolbachia and Cardinium. Our final dataset contained data from more than 117 000 arthropods (over 2500 species) screened for Wolbachia and more than 18 000 arthropods (over 800 species) screened for Cardinium, including samples from 137 different countries, with mean temperatures varying from -6.5 to 29.2°C. In insects and relatives, Cardinium infection showed a clear and consistent tendency to increase with temperature. For Wolbachia, a tendency to increase with temperature in temperate climates is counteracted by reduced prevalence in the tropics, resulting in a weak negative trend overall. We discuss the implications of these results for natural and introduced symbionts in regions affected by climate change.}, } @article {pmid31430401, year = {2019}, author = {Mathé-Hubert, H and Kaech, H and Hertaeg, C and Jaenike, J and Vorburger, C}, title = {Nonrandom associations of maternally transmitted symbionts in insects: The roles of drift versus biased cotransmission and selection.}, journal = {Molecular ecology}, volume = {28}, number = {24}, pages = {5330-5346}, doi = {10.1111/mec.15206}, pmid = {31430401}, issn = {1365-294X}, mesh = {Animals ; Aphids/*genetics/microbiology ; Bayes Theorem ; Drosophila/genetics/microbiology ; Gene Transfer, Horizontal/genetics ; Maternal Inheritance/genetics ; Microbiota/genetics ; Phylogeny ; Spiroplasma/*genetics ; Symbiosis/*genetics ; Wolbachia/*genetics ; }, abstract = {Virtually all higher organisms form holobionts with associated microbiota. To understand the biology of holobionts we need to know how species assemble and interact. Controlled experiments are suited to study interactions between particular symbionts, but they only accommodate a tiny portion of the diversity within each species. Alternatively, interactions can be inferred by testing if associations among symbionts in the field are more or less frequent than expected under random assortment. However, random assortment may not be a valid null hypothesis for maternally transmitted symbionts since drift alone can result in associations. Here, we analyse a European field survey of endosymbionts in pea aphids (Acyrthosiphon pisum), confirming that symbiont associations are pervasive. To interpret them, we develop a model simulating the effect of drift on symbiont associations. We show that drift induces apparently nonrandom assortment, even though horizontal transmissions and maternal transmission failures tend to randomise symbiont associations. We also use this model in the approximate Bayesian computation framework to revisit the association between Spiroplasma and Wolbachia in Drosophila neotestacea. New field data reported here reveal that this association has disappeared in the investigated location, yet a significant interaction between Spiroplasma and Wolbachia can still be inferred. Our study confirms that negative and positive associations are pervasive and often induced by symbiont-symbiont interactions. Nevertheless, some associations are also likely to be driven by drift. This possibility needs to be considered when performing such analyses, and our model is helpful for this purpose.}, } @article {pmid31425852, year = {2019}, author = {Huang, HJ and Cui, JR and Chen, J and Bing, XL and Hong, XY}, title = {Proteomic analysis of Laodelphax striatellus gonads reveals proteins that may manipulate host reproduction by Wolbachia.}, journal = {Insect biochemistry and molecular biology}, volume = {113}, number = {}, pages = {103211}, doi = {10.1016/j.ibmb.2019.103211}, pmid = {31425852}, issn = {1879-0240}, mesh = {Animals ; Gonads/growth & development/microbiology ; Hemiptera/genetics/growth & development/*physiology ; Insect Proteins/*genetics/metabolism ; Male ; Nymph/genetics/growth & development ; Proteome ; Proteomics ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular bacteria that manipulate host reproduction by several mechanisms including cytoplasmic incompatibility (CI). However, the underlying mechanisms of Wolbachia-induced CI are not entirely clear. Here, we monitored the Wolbachia distribution in the male gonads of the small brown planthopper (Laodelphax striatellus, SBPH) at different development stages, and investigated the influence of Wolbachia on male gonads by a quantitative proteomic analysis. A total of 276 differentially expressed proteins were identified, with the majority of them participating in metabolism, modification, and reproduction. Knocking down the expression of outer dense fiber protein (ODFP) and venom allergen 5-like (VA5L) showed decreased egg reproduction, and these two genes might be responsible for Wolbachia improved fecundity in infected L. striatellus; whereas knocking down the expression of cytosol amino-peptidase-like (CAL) significantly decreased the egg hatch rate in Wolbachia-uninfected L. striatellus, but not in the Wolbachia-infected one. Considering that the mRNA/protein level of CAL was downregulated by Wolbachia infection and dsCAL treatment closely mimicked Wolbachia-induced CI, we presumed that CAL might be one of the factors determining the CI phenotype.}, } @article {pmid31425566, year = {2019}, author = {Doremus, MR and Kelly, SE and Hunter, MS}, title = {Exposure to opposing temperature extremes causes comparable effects on Cardinium density but contrasting effects on Cardinium-induced cytoplasmic incompatibility.}, journal = {PLoS pathogens}, volume = {15}, number = {8}, pages = {e1008022}, pmid = {31425566}, issn = {1553-7374}, mesh = {Animals ; Bacteroidetes/*physiology ; *Cold Temperature ; *Host-Pathogen Interactions ; *Hot Temperature ; *Reproduction ; Symbiosis ; Wasps/*microbiology ; }, abstract = {Terrestrial arthropods, including insects, commonly harbor maternally inherited intracellular symbionts that confer benefits to the host or manipulate host reproduction to favor infected female progeny. These symbionts may be especially vulnerable to thermal stress, potentially leading to destabilization of the symbiosis and imposing costs to the host. For example, increased temperatures can reduce the density of a common reproductive manipulator, Wolbachia, and the strength of its crossing incompatibility (cytoplasmic incompatibility, or CI) phenotype. Another manipulative symbiont, Cardinium hertigii, infects ~ 6-10% of Arthropods, and also can induce CI, but there is little homology between the molecular mechanisms of CI induced by Cardinium and Wolbachia. Here we investigated whether temperature disrupts the CI phenotype of Cardinium in a parasitic wasp host, Encarsia suzannae. We examined the effects of both warm (32°C day/ 29°C night) and cool (20°C day/ 17°C night) temperatures on Cardinium CI and found that both types of temperature stress modified aspects of this symbiosis. Warm temperatures reduced symbiont density, pupal developmental time, vertical transmission rate, and the strength of both CI modification and rescue. Cool temperatures also reduced symbiont density, however this resulted in stronger CI, likely due to cool temperatures prolonging the host pupal stage. The opposing effects of cool and warm-mediated reductions in symbiont density on the resulting CI phenotype indicates that CI strength may be independent of density in this system. Temperature stress also modified the CI phenotype only if it occurred during the pupal stage, highlighting the likely importance of this stage for CI induction in this symbiosis.}, } @article {pmid31417112, year = {2019}, author = {Lorenzo-Carballa, MO and Torres-Cambas, Y and Heaton, K and Hurst, GDD and Charlat, S and Sherratt, TN and Van Gossum, H and Cordero-Rivera, A and Beatty, CD}, title = {Widespread Wolbachia infection in an insular radiation of damselflies (Odonata, Coenagrionidae).}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {11933}, pmid = {31417112}, issn = {2045-2322}, mesh = {Alleles ; Animals ; Bayes Theorem ; *Biological Evolution ; Fiji ; Geography ; Gram-Negative Bacterial Infections/*microbiology ; Host-Pathogen Interactions ; Multilocus Sequence Typing ; Odonata/*microbiology ; Phylogeny ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachia is one of the most common endosymbionts found infecting arthropods. Theory predicts symbionts like Wolbachia will be more common in species radiations, as host shift events occur with greatest frequency between closely related species. Further, the presence of Wolbachia itself may engender reproductive isolation, and promote speciation of their hosts. Here we screened 178 individuals belonging to 30 species of the damselfly genera Nesobasis and Melanesobasis - species radiations endemic to the Fiji archipelago in the South Pacific - for Wolbachia, using multilocus sequence typing to characterize bacterial strains. Incidence of Wolbachia was 71% in Nesobasis and 40% in Melanesobasis, and prevalence was also high, with an average of 88% in the Nesobasis species screened. We identified a total of 25 Wolbachia strains, belonging to supergroups A, B and F, with some epidemic strains present in multiple species. The occurrence of Wolbachia in both males and females, and the similar global prevalence found in both sexes rules out any strong effect of Wolbachia on the primary sex-ratio, but are compatible with the phenotype of cytoplasmic incompatibility. Nesobasis has higher species richness than most endemic island damselfly genera, and we discuss the potential for endosymbiont-mediated speciation within this group.}, } @article {pmid31416478, year = {2019}, author = {Gonçalves, DDS and Hooker, DJ and Dong, Y and Baran, N and Kyrylos, P and Iturbe-Ormaetxe, I and Simmons, CP and O'Neill, SL}, title = {Detecting wMel Wolbachia in field-collected Aedes aegypti mosquitoes using loop-mediated isothermal amplification (LAMP).}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {404}, pmid = {31416478}, issn = {1756-3305}, mesh = {Aedes/*microbiology ; Animals ; Colorimetry ; DNA Primers ; Genes, Bacterial ; Mosquito Vectors/microbiology ; *Nucleic Acid Amplification Techniques ; Queensland ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: The World Mosquito Program uses Wolbachia pipientis for the biocontrol of arboviruses transmitted by Aedes aegypti mosquitoes. Diagnostic testing for Wolbachia in laboratory colonies and in field-caught mosquito populations has typically employed PCR. New, simpler methods to diagnose Wolbachia infection in mosquitoes are required for large-scale operational use.

METHODS: Field-collected Ae. aegypti mosquitoes from North Queensland were tested using primers designed to detect the Wolbachia wsp gene, specific to the strain wMel. The results were analysed by colour change in the reaction mix. Furthermore, to confirm the efficiency of the LAMP assay, the results were compared to the gold-standard qPCR test.

RESULTS: A novel loop-mediated isothermal amplification (LAMP) colorimetric test for the wMel strain of Wolbachia was designed, developed and validated for use in a high-throughput setting. Against the standard qPCR test, the analytical sensitivity, specificity and diagnostic metrics were: sensitivity (99.6%), specificity (92.2%), positive predictive value (97.08%) and negative predictive value (99.30%).

CONCLUSIONS: We describe an alternative, novel and high-throughput method for diagnosing wMel Wolbachia infections in mosquitoes. This assay should support Wolbachia surveillance in both laboratory and field populations of Ae. aegypti.}, } @article {pmid31411007, year = {2020}, author = {Zhu, YX and Song, ZR and Song, YL and Zhao, DS and Hong, XY}, title = {The microbiota in spider mite feces potentially reflects intestinal bacterial communities in the host.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {859-868}, doi = {10.1111/1744-7917.12716}, pmid = {31411007}, issn = {1744-7917}, support = {2019M651864//China Postdoctoral Science Foundation/ ; 31672035//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; 31901888//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Bacteria/*isolation & purification ; Feces/microbiology ; Gastrointestinal Microbiome ; *Host Microbial Interactions ; Microbiota/*physiology ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; *Symbiosis ; Tetranychidae/*microbiology ; }, abstract = {Microorganisms provide many physiological functions to herbivorous hosts. Spider mites (genus Tetranychus) are important agricultural pests throughout the world; however, the composition of the spider mite microbial community, especially gut microbiome, remains unclear. Here, we investigated the bacterial community in five spider mite species and their associated feces by deep sequencing of the 16S rRNA gene. The composition of the bacterial community was significantly different among the five prevalent spider mite species, and some bacterial symbionts showed host-species specificity. Moreover, the abundance of the bacterial community in spider mite feces was significantly higher than that in the corresponding spider mite samples. However, Flavobacterium was detected in all samples, and represent a "core microbiome". Remarkably, the maternally inherited endosymbiont Wolbachia was detected in both spider mite and feces. Overall, these results offer insight into the complex community of symbionts in spider mites, and give a new direction for future studies.}, } @article {pmid31409391, year = {2019}, author = {Lucchetti, C and Genchi, M and Venco, L and Menozzi, A and Serventi, P and Bertini, S and Bazzocchi, C and Kramer, LH and Vismarra, A}, title = {Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {401}, pmid = {31409391}, issn = {1756-3305}, mesh = {ATP-Binding Cassette Transporters/*genetics ; Animals ; Dirofilaria immitis/*drug effects/*genetics ; Dirofilariasis/parasitology ; Dogs ; Doxycycline/*pharmacology ; Drug Combinations ; Female ; Ivermectin/*pharmacology ; Male ; }, abstract = {BACKGROUND: Combination doxycycline/macrocyclic lactone (ML) protocols have been shown to provide a more rapid adulticidal and microfilaricidal effect than either MLs or doxycycline alone, although female worms were reported to have a higher tolerance to treatments compared to male worms. The present study aimed to evaluate how ABC transporters may be involved in the synergic effect of the combination treatment. Adult worms of D. immitis were treated in vitro for 24 hours with doxycycline (DOXY), ivermectin (IVM) and a combination of both, and changes in the modulation of ABC transporter genes were measured. Levels of doxycycline inside different treatment media, post-treatment, were determined through HPLC analysis.

RESULTS: Quantitative RT-PCR analysis showed the presence of changes in the modulation of ABC transporter genes evaluated in this study. In particular, in female worms, the combination treatment induced a substantial increase in gene expressions, especially of Dim-pgp-10 and Dim-haf-4; whereas in male worms, the greatest increase in gene expression was observed for Dim-pgp-10 and Dim-pgp-11 when treated with DMSO + IVM and DMSO + DOXY/IVM. HPLC analysis of the DOXY concentrations in the media after in vitro treatments of male worms showed a slight difference between the DMSO + DOXY samples and the combination (DMSO + DOXY + IVM), while no difference was observed among females.

CONCLUSIONS: Further studies are required to explain whether the modulation of cellular efflux plays a role, even partially, in the adulticide effect of doxycycline/macrocyclic lactone combinations in heartworm-infected dogs. To the authors' knowledge, this is the first study to evaluate P-gp expression in adult D. immitis.}, } @article {pmid31407021, year = {2020}, author = {Bockoven, AA and Bondy, EC and Flores, MJ and Kelly, SE and Ravenscraft, AM and Hunter, MS}, title = {What Goes Up Might Come Down: the Spectacular Spread of an Endosymbiont Is Followed by Its Decline a Decade Later.}, journal = {Microbial ecology}, volume = {79}, number = {2}, pages = {482-494}, pmid = {31407021}, issn = {1432-184X}, support = {DEB-1020460//Division of Environmental Biology/ ; IOS-1256905//Division of Integrative Organismal Systems/ ; }, mesh = {Animals ; Arizona ; Genetic Fitness ; Hemiptera/genetics/*microbiology/*physiology ; *Microbiota ; Rickettsia/*physiology ; Sex Ratio ; *Symbiosis ; }, abstract = {Facultative, intracellular bacterial symbionts of arthropods may dramatically affect host biology and reproduction. The length of these symbiont-host associations may be thousands to millions of years, and while symbiont loss is predicted, there have been very few observations of a decline of symbiont infection rates. In a population of the sweet potato whitefly species (Bemisia tabaci MEAM1) in Arizona, USA, we documented the frequency decline of a strain of Rickettsia in the Rickettsia bellii clade from near-fixation in 2011 to 36% of whiteflies infected in 2017. In previous studies, Rickettsia had been shown to increase from 1 to 97% from 2000 to 2006 and remained at high frequency for at least five years. At that time, Rickettsia infection was associated with both fitness benefits and female bias. In the current study, we established matrilines of whiteflies from the field (2016, Rickettsia infection frequency = 58%) and studied (a) Rickettsia vertical transmission, (b) fitness and sex ratios associated with Rickettsia infection, (c) symbiont titer, and (d) bacterial communities within whiteflies. The vertical transmission rate was high, approximately 98%. Rickettsia infection in the matrilines was not associated with fitness benefits or sex ratio bias and appeared to be slightly costly, as more Rickettsia-infected individuals produced non-hatching eggs. Overall, the titer of Rickettsia in the matrilines was lower in 2016 than in the whiteflies collected in 2011, but the titer distribution appeared bimodal, with high- and low-titer lines, and constancy of the average titer within lines over three generations. We found neither association between Rickettsia titer and fitness benefits or sex ratio bias nor evidence that Rickettsia was replaced by another secondary symbiont. The change in the interaction between symbiont and host in 2016 whiteflies may explain the drop in symbiont frequency we observed.}, } @article {pmid31397383, year = {2019}, author = {Anderson, ML and Rustin, RC and Eremeeva, ME}, title = {Pilot survey of mosquitoes (Diptera: Culicidae) from southeastern Georgia, USA for Wolbachia and Rickettsia felis (Rickettsiales: Rickettsiaceae).}, journal = {Journal of vector borne diseases}, volume = {56}, number = {2}, pages = {92-97}, doi = {10.4103/0972-9062.263714}, pmid = {31397383}, issn = {0972-9062}, mesh = {Animals ; Culicidae/*microbiology ; DNA, Bacterial/genetics ; Female ; Genotype ; Georgia ; Male ; Pilot Projects ; RNA, Ribosomal, 16S ; Rickettsiaceae/genetics/*isolation & purification ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND & OBJECTIVES: Mosquito surveillance is one of the critical functions of local health departments, particularly in the context of outbreaks of severe mosquito-borne viral infections. Unfortunately, some viral and parasitic infections transmitted by mosquitoes, manifests non-specific clinical symptoms which may actually be of rickettsial etiology, including Rickettsia felis infections. This study tested the hypothesis that mosquitoes from southeastern Georgia, USA may be infected with Rickettsia felis and Wolbachia, an endosymbiotic bacterium of the order Rickettsiales.

METHODS: Specimens of the five most common mosquito species occurring in the region were collected using gravid and light-traps and identified using morphological keys. Mosquitoes were then pooled by species, sex, trap and collection site and their DNA was extracted. Molecular methods were used to confirm mosquito identification, and presence of Wolbachia and R. felis.

RESULTS: Wolbachia DNA was detected in 90.8% of the mosquito pools tested, which included 98% pools of Cx. quinquefasciatus Say (Diptera: Culicidae), 95% pools of Ae. albopictus Skuse (Diptera: Culicidae), and 66.7% of pools of Cx. pipiens complex. Samples of An. punctipennis Say (Diptera: Culicidae) and An. crucians Wiedemann (Diptera: Culicidae) were tested negative for Wolbachia DNA. Three genotypes of Wolbachia sp. belonging to Group A (1 type) and Group B (2 types) were identified. DNA of R. felis was not found in any pool of mosquitoes tested.

This study provides a pilot data on the high presence of Wolbachia in Cx. quinque-fasciatus and Ae. albopictus mosquitoes prevalent in the study region. Whether the high prevalence of Wolbachia and its genetic diversity in mosquitoes affects the mosquitoes' susceptibility to R. felis infection in Georgia will need further evaluation.}, } @article {pmid31396100, year = {2019}, author = {Liu, L and Zhang, KJ and Rong, X and Li, YY and Liu, H}, title = {Identification of Wolbachia-Responsive miRNAs in the Small Brown Planthopper, Laodelphax striatellus.}, journal = {Frontiers in physiology}, volume = {10}, number = {}, pages = {928}, pmid = {31396100}, issn = {1664-042X}, abstract = {Laodelphax striatellus is naturally infected with the Wolbachia strain wStri, which induces strong cytoplasmic incompatibility of its host. MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that play a critical role in the regulation of gene expression at post-transcriptional level in various biological processes. Despite various studies reporting that Wolbachia affects the miRNA expression of their hosts, the molecular mechanism underlying interactions between Wolbachia and their host miRNAs has not been well understood. In order to better understand the impact of Wolbachia infection on its host, we investigated the differentially expressed miRNAs between Wolbachia-infected and Wolbachia-uninfected strains of L. striatellus. Compared with uninfected strains, Wolbachia infection resulted in up-regulation of 18 miRNAs and down-regulation of 6 miRNAs in male, while 25 miRNAs were up-regulated and 15 miRNAs were down-regulated in female. The target genes of these differentially expressed miRNAs involved in immune response regulation, reproduction, redox homeostasis and ecdysteroidogenesis were also annotated in both sexes. We further verified the expression of several significantly differentially expressed miRNAs and their predicted target genes by qRT-PCR method. The results suggested that Wolbachia appears to reduce the expression of genes related to fertility in males and increase the expression of genes related to fecundity in females. At the same time, Wolbachia may enhance the expression of immune-related genes in both sexes. All of the results in this study may be helpful in further exploration of the molecular mechanisms by which Wolbachia affects on its hosts.}, } @article {pmid31384012, year = {2019}, author = {Klinges, JG and Rosales, SM and McMinds, R and Shaver, EC and Shantz, AA and Peters, EC and Eitel, M and Wörheide, G and Sharp, KH and Burkepile, DE and Silliman, BR and Vega Thurber, RL}, title = {Phylogenetic, genomic, and biogeographic characterization of a novel and ubiquitous marine invertebrate-associated Rickettsiales parasite, Candidatus Aquarickettsia rohweri, gen. nov., sp. nov.}, journal = {The ISME journal}, volume = {13}, number = {12}, pages = {2938-2953}, pmid = {31384012}, issn = {1751-7370}, mesh = {Animals ; Aquatic Organisms/*microbiology ; Genome, Bacterial ; Genomics ; Gram-Negative Bacterial Infections/microbiology/*veterinary ; Invertebrates/*microbiology ; Parasites/classification/genetics/isolation & purification ; *Phylogeny ; Rickettsiales/classification/genetics/*isolation & purification ; }, abstract = {Bacterial symbionts are integral to the health and homeostasis of invertebrate hosts. Notably, members of the Rickettsiales genus Wolbachia influence several aspects of the fitness and evolution of their terrestrial hosts, but few analogous partnerships have been found in marine systems. We report here the genome, phylogenetics, and biogeography of a ubiquitous and novel Rickettsiales species that primarily associates with marine organisms. We previously showed that this bacterium was found in scleractinian corals, responds to nutrient exposure, and is associated with reduced host growth and increased mortality. This bacterium, like other Rickettsiales, has a reduced genome indicative of a parasitic lifestyle. Phylogenetic analysis places this Rickettsiales within a new genus we define as "Candidatus Aquarickettsia." Using data from the Earth Microbiome Project and SRA databases, we also demonstrate that members of "Ca. Aquarickettsia" are found globally in dozens of invertebrate lineages. The coral-associated "Candidatus A. rohweri" is the first finished genome in this new clade. "Ca. A. rohweri" lacks genes to synthesize most sugars and amino acids but possesses several genes linked to pathogenicity including Tlc, an antiporter that exchanges host ATP for ADP, and a complete Type IV secretion system. Despite its inability to metabolize nitrogen, "Ca. A. rohweri" possesses the NtrY-NtrX two-component system involved in sensing and responding to extracellular nitrogen. Given these data, along with visualization of the parasite in host tissues, we hypothesize that "Ca. A. rohweri" reduces coral health by consuming host nutrients and energy, thus weakening and eventually killing host cells. Last, we hypothesize that nutrient enrichment, which is increasingly common on coral reefs, encourages unrestricted growth of "Ca. A. rohweri" in its host by providing abundant N-rich metabolites to be scavenged.}, } @article {pmid31381563, year = {2019}, author = {Hübner, MP and Koschel, M and Struever, D and Nikolov, V and Frohberger, SJ and Ehrens, A and Fendler, M and Johannes, I and von Geldern, TW and Marsh, K and Turner, JD and Taylor, MJ and Ward, SA and Pfarr, K and Kempf, DJ and Hoerauf, A}, title = {In vivo kinetics of Wolbachia depletion by ABBV-4083 in L. sigmodontis adult worms and microfilariae.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {8}, pages = {e0007636}, pmid = {31381563}, issn = {1935-2735}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Doxycycline/pharmacology ; Female ; Filariasis ; Filarioidea/drug effects/*microbiology ; Gerbillinae ; Kinetics ; Mice ; Mice, Inbred BALB C ; Microfilariae/drug effects/embryology/*microbiology ; Models, Animal ; Wolbachia/*drug effects/*physiology ; }, abstract = {Depletion of Wolbachia endosymbionts of human pathogenic filariae using 4-6 weeks of doxycycline treatment can lead to permanent sterilization and adult filarial death. We investigated the anti-Wolbachia drug candidate ABBV-4083 in the Litomosoides sigmodontis rodent model to determine Wolbachia depletion kinetics with different regimens. Wolbachia reduction occurred in mice as early as 3 days after the initiation of ABBV-4083 treatment and continued throughout a 10-day treatment period. Importantly, Wolbachia levels continued to decline after a 5-day-treatment from 91.5% to 99.9% during a 3-week washout period. In jirds, two weeks of ABBV-4083 treatment (100mg/kg once-per-day) caused a >99.9% Wolbachia depletion in female adult worms, and the kinetics of Wolbachia depletion were recapitulated in peripheral blood microfilariae. Similar to Wolbachia depletion, inhibition of embryogenesis was time-dependent in ABBV-4083-treated jirds, leading to a complete lack of late embryonic stages (stretched microfilariae) and lack of peripheral microfilariae in 5/6 ABBV-4083-treated jirds by 14 weeks after treatment. Twice daily treatment in comparison to once daily treatment with ABBV-4083 did not significantly improve Wolbachia depletion. Moreover, up to 4 nonconsecutive daily treatments within a 14-dose regimen did not significantly erode Wolbachia depletion. Within the limitations of an animal model that does not fully recapitulate human filarial disease, our studies suggest that Wolbachia depletion should be assessed clinically no earlier than 3-4 weeks after the end of treatment, and that Wolbachia depletion in microfilariae may be a viable surrogate marker for the depletion within adult worms. Furthermore, strict daily adherence to the dosing regimen with anti-Wolbachia candidates may not be required, provided that the full regimen is subsequently completed.}, } @article {pmid31380084, year = {2019}, author = {Yoshida, K and Sanada-Morimura, S and Huang, SH and Tokuda, M}, title = {Influences of two coexisting endosymbionts, CI-inducing Wolbachia and male-killing Spiroplasma, on the performance of their host Laodelphax striatellus (Hemiptera: Delphacidae).}, journal = {Ecology and evolution}, volume = {9}, number = {14}, pages = {8214-8224}, pmid = {31380084}, issn = {2045-7758}, abstract = {The small brown planthopper Laodelphax striatellus (Hemiptera: Delphacidae) is reported to have the endosymbiont Wolbachia, which shows a strong cytoplasmic incompatibility (CI) between infected males and uninfected females. In the 2000s, female-biased L. striatellus populations were found in Taiwan, and this sex ratio distortion was the result of male-killing induced by the infection of another endosymbiont, Spiroplasma. Spiroplasma infection is considered to negatively affect both L. striatellus and Wolbachia because the male-killing halves the offspring of L. striatellus and hinders the spread of Wolbachia infection via CI. Spiroplasma could have traits that increase the fitness of infected L. striatellus and/or coexisting organisms because the coinfection rates of Wolbachia and Spiroplasma were rather high in some areas. In this study, we investigated the influences of the infection of these two endosymbionts on the development, reproduction, and insecticide resistance of L. striatellus in the laboratory. Our results show that the single-infection state of Spiroplasma had a negative influence on the fertility of L. striatellus, while the double-infection state had no significant influence. At late nymphal and adult stages, the abundance of Spiroplasma was lower in the double-infection state than in the single-infection state. In the double-infection state, the reduction of Spiroplasma density may be caused by competition between the two endosymbionts, and the negative influence of Spiroplasma on the fertility of host may be relieved. The resistance of L. striatellus to four insecticides was compared among different infection states of endosymbionts, but Spiroplasma infection did not contribute to increase insecticide resistance. Because positive influences of Spiroplasma infection were not found in terms of the development, reproduction, and insecticide resistance of L. striatellus, other factors improving the fitness of Spiroplasma-infected L. striatellus may be related to the high frequency of double infection in some L. striatellus populations.}, } @article {pmid31380078, year = {2019}, author = {Hu, G and Zhang, L and Yun, Y and Peng, Y}, title = {Taking insight into the gut microbiota of three spider species: No characteristic symbiont was found corresponding to the special feeding style of spiders.}, journal = {Ecology and evolution}, volume = {9}, number = {14}, pages = {8146-8156}, pmid = {31380078}, issn = {2045-7758}, abstract = {Microorganisms in insect guts have been recognized as having a great impact on their hosts' nutrition, health, and behavior. Spiders are important natural enemies of pests, and the composition of the gut microbiota of spiders remains unclear. Will the bacterial taxa in spiders be same as the bacterial taxa in insects, and what are the potential functions of the gut bacteria in spiders? To gain insight into the composition of the gut bacteria in spiders and their potential function, we collected three spider species, Pardosa laura, Pardosa astrigera, and Nurscia albofasciata, in the field, and high-throughput sequencing of the 16S rRNA V3 and V4 regions was used to investigate the diversity of gut microbiota across the three spider species. A total of 23 phyla and 150 families were identified in these three spider species. The dominant bacterial phylum across all samples was Proteobacteria. Burkholderia, Ralstonia, Ochrobactrum, Providencia, Acinetobacter, Proteus, and Rhodoplanes were the dominant genera in the guts of the three spider species. The relative abundances of Wolbachia and Rickettsiella detected in N. albofasciata were significantly higher than those in the other two spider species. The relative abundance of Thermus, Amycolatopsis, Lactococcus, Acinetobacter Microbacterium, and Koribacter detected in spider gut was different among the three spider species. Biomolecular interaction networks indicated that the microbiota in the guts had complex interactions. The results of this study also suggested that at the genus level, some of the gut bacteria taxa in the three spider species were the same as the bacteria in insect guts.}, } @article {pmid31380036, year = {2019}, author = {Su, Q and Wang, X and Ilyas, N and Zhang, F and Yun, Y and Jian, C and Peng, Y}, title = {Combined effects of elevated CO2 concentration and Wolbachia on Hylyphantes graminicola (Araneae: Linyphiidae).}, journal = {Ecology and evolution}, volume = {9}, number = {12}, pages = {7112-7121}, pmid = {31380036}, issn = {2045-7758}, abstract = {The increasing concentration of carbon dioxide in atmosphere is not only a major cause of global warming, but it also adversely affects the ecological diversity of invertebrates. This study was conducted to evaluate the effect of elevated CO2 concentration (ambient, 400 ppm and high, 800 ppm) and Wolbachia (Wolbachia-infected, W[+] and Wolbachia-uninfected, W[-]) on Hylyphantes graminicola. The total survival rate, developmental duration, carapace width and length, body weight, sex ratio, net reproductive rate, nutrition content, and enzyme activity in H. graminicola were examined under four treatments: W[-] 400 ppm, W[-] 800 ppm, W[+] 400 ppm, and W[+] 800 ppm. Results showed that Wolbachia-infected spiders had significantly decreased the total developmental duration. Different instars showed variations up to some extent, but no obvious effect was found under elevated CO2 concentration. Total survival rate, sex ratio, and net reproductive rate were not affected by elevated CO2 concentration or Wolbachia infection. The carapace width of Wolbachia-uninfected spiders decreased significantly under elevated CO2 concentration, while the width, length and weight were not significantly affected in Wolbachia-infected spiders reared at ambient CO2 concentration. The levels of protein, specific activities of peroxidase, and amylase were significantly increased under elevated CO2 concentration or Wolbachia-infected spiders, while the total amino content was only increased in Wolbachia-infected spiders. Thus, our current finding suggested that elevated CO2 concentration and Wolbachia enhance nutrient contents and enzyme activity of H. graminicola and decrease development duration hence explore the interactive effects of factors which were responsible for reproduction regulation, but it also gives a theoretical direction for spider's protection in such a dynamic environment. Increased activities of enzymes and nutrients caused by Wolbachia infection aids for better survival of H. graminicola under stress.}, } @article {pmid31375435, year = {2019}, author = {Dimopoulos, G}, title = {Combining Sterile and Incompatible Insect Techniques for Aedes albopictus Suppression.}, journal = {Trends in parasitology}, volume = {35}, number = {9}, pages = {671-673}, doi = {10.1016/j.pt.2019.07.006}, pmid = {31375435}, issn = {1471-5007}, support = {R01 AI141532/AI/NIAID NIH HHS/United States ; R01 AI122743/AI/NIAID NIH HHS/United States ; R21 AI131574/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes ; Animals ; *Dengue ; *Infertility ; Male ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Traditional control strategies are failing to contain Aedes albopictus as an emerging major vector for dengue. A combination of approaches (Zheng et al., Nature, 2019) involving an artificial triple Wolbachia superinfection and low-dose irradiation enabled mass production of adult sterile males for release. The resulting suppression of field populations suggests feasibility for area-wide vector control.}, } @article {pmid31369548, year = {2019}, author = {Schneider, DI and Saarman, N and Onyango, MG and Hyseni, C and Opiro, R and Echodu, R and O'Neill, M and Bloch, D and Vigneron, A and Johnson, TJ and Dion, K and Weiss, BL and Opiyo, E and Caccone, A and Aksoy, S}, title = {Spatio-temporal distribution of Spiroplasma infections in the tsetse fly (Glossina fuscipes fuscipes) in northern Uganda.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {8}, pages = {e0007340}, pmid = {31369548}, issn = {1935-2735}, support = {D43 TW007391/TW/FIC NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; R01 AI139525/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Coinfection ; DNA, Ribosomal/genetics ; Female ; Gram-Negative Bacterial Infections/*microbiology/*veterinary ; Insect Vectors/*microbiology/parasitology ; Male ; Prevalence ; Spiroplasma/genetics/*pathogenicity/physiology ; Symbiosis ; Trypanosoma ; Tsetse Flies/*microbiology/parasitology ; Uganda ; Wolbachia ; }, abstract = {Tsetse flies (Glossina spp.) are vectors of parasitic trypanosomes, which cause human (HAT) and animal African trypanosomiasis (AAT) in sub-Saharan Africa. In Uganda, Glossina fuscipes fuscipes (Gff) is the main vector of HAT, where it transmits Gambiense disease in the northwest and Rhodesiense disease in central, southeast and western regions. Endosymbionts can influence transmission efficiency of parasites through their insect vectors via conferring a protective effect against the parasite. It is known that the bacterium Spiroplasma is capable of protecting its Drosophila host from infection with a parasitic nematode. This endosymbiont can also impact its host's population structure via altering host reproductive traits. Here, we used field collections across 26 different Gff sampling sites in northern and western Uganda to investigate the association of Spiroplasma with geographic origin, seasonal conditions, Gff genetic background and sex, and trypanosome infection status. We also investigated the influence of Spiroplasma on Gff vector competence to trypanosome infections under laboratory conditions. Generalized linear models (GLM) showed that Spiroplasma probability was correlated with the geographic origin of Gff host and with the season of collection, with higher prevalence found in flies within the Albert Nile (0.42 vs 0.16) and Achwa River (0.36 vs 0.08) watersheds and with higher prevalence detected in flies collected in the intermediate than wet season. In contrast, there was no significant correlation of Spiroplasma prevalence with Gff host genetic background or sex once geographic origin was accounted for in generalized linear models. Additionally, we found a potential negative correlation of Spiroplasma with trypanosome infection, with only 2% of Spiroplasma infected flies harboring trypanosome co-infections. We also found that in a laboratory line of Gff, parasitic trypanosomes are less likely to colonize the midgut in individuals that harbor Spiroplasma infection. These results indicate that Spiroplasma infections in tsetse may be maintained by not only maternal but also via horizontal transmission routes, and Spiroplasma infections may also have important effects on trypanosome transmission efficiency of the host tsetse. Potential functional effects of Spiroplasma infection in Gff could have impacts on vector control approaches to reduce trypanosome infections.}, } @article {pmid31362350, year = {2019}, author = {Satjawongvanit, H and Phumee, A and Tiawsirisup, S and Sungpradit, S and Brownell, N and Siriyasatien, P and Preativatanyou, K}, title = {Molecular Analysis of Canine Filaria and Its Wolbachia Endosymbionts in Domestic Dogs Collected from Two Animal University Hospitals in Bangkok Metropolitan Region, Thailand.}, journal = {Pathogens (Basel, Switzerland)}, volume = {8}, number = {3}, pages = {}, pmid = {31362350}, issn = {2076-0817}, abstract = {Canine filariasis is caused by several nematode species, such as Dirofilaria immitis, Dirofilaria repens, Brugia pahangi, Brugia malayi, and Acanthocheilonema reconditum. Zoonotic filariasis is one of the world's neglected tropical diseases. Since 2000, the World Health Organization (WHO) has promoted a global filarial eradication program to eliminate filariasis by 2020. Apart from vector control strategies, the infection control of reservoir hosts is necessary for more effective filariasis control. In addition, many studies have reported that Wolbachia is necessary for the development, reproduction, and survival of the filarial nematode. Consequently, the use of antibiotics to kill Wolbachia in nematodes has now become an alternative strategy to control filariasis. Previously, a case of subconjunctival dirofilariasis caused by Dirofilaria spp. has been reported in a woman who resides in the center of Bangkok, Thailand. Therefore, our study aimed to principally demonstrate the presence of filarial nematodes and Wolbachia bacteria in blood collected from domestic dogs from the Bangkok Metropolitan Region, Thailand. A total of 57 blood samples from dogs with suspected dirofilariasis who had visited veterinary clinics in Bangkok were collected. The investigations for the presence of microfilaria were carried out by using both microscopic and molecular examinations. PCR was used as the molecular detection method for the filarial nematodes based on the COI and ITS1 regions. The demonstration of Wolbachia was performed using PCR to amplify the FtsZ gene. All positive samples by PCR were then cloned and sequenced. The results showed that the filarial nematodes were detected in 16 samples (28.07%) using microscopic examinations. The molecular detection of filarial species using COI-PCR revealed that 50 samples (87.72%) were positive; these consisted of 33 (57.89%), 13 (22.81%), and 4 (7.02%) samples for D. immitis, B. pahangi, and B. malayi, respectively. While the ITS1-PCR showed that 41 samples (71.93%) were positive-30 samples (52.63%) were identified as containing D. immitis and 11 samples (19.30%) were identified to have B. pahangi, whereas B. malayi was not detected. Forty-seven samples (82.45%) were positive for Wolbachia DNA and the phylogenetic tree of all positive Wolbachia was classified into the supergroup C clade. This study has established fundamental data on filariasis associated with Wolbachia infection in domestic dogs in the Bangkok Metropolitan Region. An extensive survey of dog blood samples would provide valuable epidemiologic data on potential zoonotic filariasis in Thailand. In addition, this information could be used for the future development of more effective prevention and control strategies for canine filariasis in Thailand.}, } @article {pmid31358201, year = {2019}, author = {Liu, XD and Guo, HF}, title = {Importance of endosymbionts Wolbachia and Rickettsia in insect resistance development.}, journal = {Current opinion in insect science}, volume = {33}, number = {}, pages = {84-90}, doi = {10.1016/j.cois.2019.05.003}, pmid = {31358201}, issn = {2214-5753}, mesh = {Animals ; Inactivation, Metabolic ; Insecta/*microbiology/physiology ; *Insecticide Resistance ; Insecticides/toxicity ; Rickettsia/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Endosymbionts play important roles in protecting hosts from environmental stress, such as natural enemies, heat, and toxins. Many insects are infected with the facultative nonessential endosymbionts Wolbachia and Rickettsia, which are the crux in this review, although other relevant symbiont genera will also be treated. Insecticide resistance of hosts can be related to infections with Wolbachia and Rickettsia. These endosymbionts commonly increase host susceptibility to chemical insecticides, but cases of increased resistance also exist. The symbiont-mediated insecticide resistance/susceptibility varies with species of insect, species of symbiont, and chemical compound. Changes in insecticide resistance levels of insects can be associated with fluctuations in population density of endosymbionts. Effects of endosymbionts on host fitness, metabolism, immune system, and gene expression may determine how endosymbionts influence insecticide resistance. A clearer understanding of these interactions can improve our knowledge about drivers of decreasing insecticide resistance.}, } @article {pmid31351474, year = {2019}, author = {Singh, JA}, title = {Informed consent and community engagement in open field research: lessons for gene drive science.}, journal = {BMC medical ethics}, volume = {20}, number = {1}, pages = {54}, pmid = {31351474}, issn = {1472-6939}, mesh = {Animals ; *Communicable Disease Control/methods ; *Community Participation ; Community-Based Participatory Research/*ethics/methods/standards ; Culicidae/genetics ; Dengue/prevention & control ; Disease Transmission, Infectious/prevention & control ; Gene Drive Technology/*ethics/methods ; Humans ; *Informed Consent/ethics/standards ; Mosquito Control/ethics/methods ; Mosquito Vectors/genetics ; Randomized Controlled Trials as Topic/ethics ; Trust ; Wolbachia ; }, abstract = {BACKGROUND: The development of the CRISPR/Cas9 gene editing system has generated new possibilities for the use of gene drive constructs to reduce or suppress mosquito populations to levels that do not support disease transmission. Despite this prospect, social resistance to genetically modified organisms remains high. Gene drive open field research thus raises important questions regarding what is owed to those who may not consent to such research, or those could be affected by the proposed research, but whose consent is not solicited. The precise circumstances under which informed consent must be obtained, and from whom, requires careful consideration. Furthermore, appropriate engagement processes should be central to any introduction of genetically modified mosquitos in proposed target settings.

DISCUSSION: In this work, international guidance documents on informed consent and engagement are reviewed and applied to the genetically modified mosquito research context. Five analogous research endeavours that involve area-wide / open field experiments are reviewed. The approach of each in respect to the solicitation of individual informed consent and community engagement are highlighted.

CONCLUSIONS: While the solicitation of individual informed consent in host settings of gene drive field trials may not be possible or feasible in some instances, local community and stakeholder engagement will be key to building trust towards the proposed conduct of such research. In this regard, the approaches taken by investigators and sponsors of political science field research and weather modification field research should be avoided. Rather, proponents of gene drive field research should look to the Eliminate Dengue field trials, cluster randomised trials, and pragmatic clinical trials for guidance regarding how the solicitation of individual informed consent of host communities ought to be managed, and how these communities ought to be engaged.}, } @article {pmid31350319, year = {2019}, author = {Martin, E and Borucki, MK and Thissen, J and Garcia-Luna, S and Hwang, M and Wise de Valdez, M and Jaing, CJ and Hamer, GL and Frank, M}, title = {Mosquito-Borne Viruses and Insect-Specific Viruses Revealed in Field-Collected Mosquitoes by a Monitoring Tool Adapted from a Microbial Detection Array.}, journal = {Applied and environmental microbiology}, volume = {85}, number = {19}, pages = {}, pmid = {31350319}, issn = {1098-5336}, support = {K01 AI128005/AI/NIAID NIH HHS/United States ; R21 AI128953/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/virology ; Animals ; Arbovirus Infections/prevention & control ; Arboviruses/*genetics/isolation & purification ; Culex/virology ; Dengue Virus/genetics/isolation & purification ; Female ; Flavivirus/genetics/isolation & purification ; Insect Viruses/*genetics/*isolation & purification ; Limit of Detection ; Mosquito Vectors/*virology ; Oligonucleotide Array Sequence Analysis/instrumentation/*methods ; Reproducibility of Results ; Sensitivity and Specificity ; Texas ; Wolbachia/virology ; }, abstract = {Several mosquito-borne diseases affecting humans are emerging or reemerging in the United States. The early detection of pathogens in mosquito populations is essential to prevent and control the spread of these diseases. In this study, we tested the potential applicability of the Lawrence Livermore Microbial Detection Array (LLMDA) to enhance biosurveillance by detecting microbes present in Aedes aegypti, Aedes albopictus, and Culex mosquitoes, which are major vector species globally, including in Texas. The sensitivity and reproducibility of the LLMDA were tested in mosquito samples spiked with different concentrations of dengue virus (DENV), revealing a detection limit of >100 but <1,000 PFU/ml. Additionally, field-collected mosquitoes from Chicago, IL, and College Station, TX, of known infection status (West Nile virus [WNV] and Culex flavivirus [CxFLAV] positive) were tested on the LLMDA to confirm its efficiency. Mosquito field samples of unknown infection status, collected in San Antonio, TX, and the Lower Rio Grande Valley (LRGV), TX, were run on the LLMDA and further confirmed by PCR or quantitative PCR (qPCR). The analysis of the field samples with the LLMDA revealed the presence of cell-fusing agent virus (CFAV) in A. aegypti populations. Wolbachia was also detected in several of the field samples (A. albopictus and Culex spp.) by the LLMDA. Our findings demonstrated that the LLMDA can be used to detect multiple arboviruses of public health importance, including viruses that belong to the Flavivirus, Alphavirus, and Orthobunyavirus genera. Additionally, insect-specific viruses and bacteria were also detected in field-collected mosquitoes. Another strength of this array is its ability to detect multiple viruses in the same mosquito pool, allowing for the detection of cocirculating pathogens in an area and the identification of potential ecological associations between different viruses. This array can aid in the biosurveillance of mosquito-borne viruses circulating in specific geographical areas.IMPORTANCE Viruses associated with mosquitoes have made a large impact on public and veterinary health. In the United States, several viruses, including WNV, DENV, and chikungunya virus (CHIKV), are responsible for human disease. From 2015 to 2018, imported Zika cases were reported in the United States, and in 2016 to 2017, local Zika transmission occurred in the states of Texas and Florida. With globalization and a changing climate, the frequency of outbreaks linked to arboviruses will increase, revealing a need to better detect viruses in vector populations. With the capacity of the LLMDA to detect viruses, bacteria, and fungi, this study highlights its ability to broadly screen field-collected mosquitoes and contribute to the surveillance and management of arboviral diseases.}, } @article {pmid31348717, year = {2019}, author = {Speer, KA and Luetke, E and Bush, E and Sheth, B and Gerace, A and Quicksall, Z and Miyamoto, M and Dick, CW and Dittmar, K and Albury, N and Reed, DL}, title = {A Fly on the Cave Wall: Parasite Genetics Reveal Fine-scale Dispersal Patterns of Bats.}, journal = {The Journal of parasitology}, volume = {105}, number = {4}, pages = {555-566}, pmid = {31348717}, issn = {1937-2345}, mesh = {Algorithms ; Alleles ; Animal Distribution/*physiology ; Animals ; Bahamas ; Bayes Theorem ; Chiroptera/*parasitology/*physiology ; Cluster Analysis ; Consensus Sequence ; DNA, Mitochondrial/chemistry ; Diptera/genetics/microbiology/*physiology ; Ectoparasitic Infestations/parasitology/*veterinary ; Electron Transport Complex IV/chemistry/genetics ; Host-Parasite Interactions ; Markov Chains ; Sequence Alignment/veterinary ; Wolbachia/physiology ; }, abstract = {Dispersal influences the evolution and adaptation of organisms, but it can be difficult to detect. Host-specific parasites provide information about the dispersal of their hosts and may be valuable for examining host dispersal that does not result in gene flow or that has low signals of gene flow. We examined the population connectivity of the buffy flower bat, Erophylla sezekorni (Chiroptera: Phyllostomidae), and its associated obligate ectoparasite, Trichobius frequens (Diptera: Streblidae), across a narrow oceanic channel in The Bahamas that has previously been implicated as a barrier to dispersal in bats. Due to the horizontal transmission of T. frequens, we were able to test the hypothesis that bats are dispersing across this channel, but this dispersal does not result in gene flow, occurs rarely, or started occurring recently. We developed novel microsatellite markers for the family Streblidae in combination with previously developed markers for bats to genotype individuals from 4 islands in The Bahamas. We provide evidence for a single population of the host, E. sezekorni, but 2 populations of its bat flies, potentially indicating a recent reduction of gene flow in E. sezekorni, rare dispersal, or infrequent transportation of bat flies with their hosts. Despite high population differentiation in bat flies indicated by microsatellites, mitochondrial DNA shows no polymorphism, suggesting that bacterial reproductive parasites may be contributing to mitochondrial DNA sweeps. Parasites, including bat flies, provide independent information about their hosts and can be used to test hypotheses of host dispersal that may be difficult to assess using host genetics alone.}, } @article {pmid31346538, year = {2019}, author = {Khanmohammadi, M and Falak, R and Meamar, AR and Arshadi, M and Akhlaghi, L and Razmjou, E}, title = {Molecular Detection and Phylogenetic Analysis of Endosymbiont Wolbachia pipientis (Rickettsiales: Anaplasmataceae) Isolated from Dirofilaria immitis in Northwest of Iran.}, journal = {Journal of arthropod-borne diseases}, volume = {13}, number = {1}, pages = {83-93}, pmid = {31346538}, issn = {2322-1984}, abstract = {BACKGROUND: The purpose of this study was molecular detection and phylogenetic analysis of Wolbachia species of Dirofilaria immitis.

METHODS: Adult filarial nematodes were collected from the cardiovascular and pulmonary arterial systems of naturally infected dogs, which caught in different geographical areas of Meshkin Shahr in Ardabil Province, Iran, during 2017. Dirofilaria immitis genomic DNA were extracted. Phylogenetic analysis for proofing of D. immitis was carried out using cytochrome oxidase I (COI) gene. Afterward, the purified DNA was used to determine the molecular pattern of the Wolbachia surface protein (WSP) gene sequence by PCR.

RESULTS: Phylogeny and homology studies showed high consistency of the COI gene with the previously-registered sequences for D. immitis. Comparison of DNA sequences revealed no nucleotide variation between them. PCR showed that all of the collected parasites were infected with W. pipientis. The sequence of the WSP gene in Wolbachia species from D. immitis was significantly different from other species of Dirofilaria as well as other filarial species. The maximum homology was observed with the Wolbachia isolated from D. immitis. The greatest distance between WSP nucleotides of Wolbachia species found between D. immitis and those isolated from Onchocerca lupi.

CONCLUSION: PCR could be a simple but suitable method for detection of Wolbachia species. There is a pattern of host specificity between Wolbachia and Dirofilaria that can be related to ancestral evolutions. The results of this phylogenetic analysis and molecular characterization may help us for better identification of Wolbachia species and understanding of their coevolution.}, } @article {pmid31340862, year = {2019}, author = {Carvajal, TM and Hashimoto, K and Harnandika, RK and Amalin, DM and Watanabe, K}, title = {Detection of Wolbachia in field-collected Aedes aegypti mosquitoes in metropolitan Manila, Philippines.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {361}, pmid = {31340862}, issn = {1756-3305}, support = {16H05750//Japanese Society for the Promotion of Science/ ; 17H01624//Japanese Society for the Promotion of Science/ ; 17K18906//Japanese Society for the Promotion of Science/ ; Bilateral Joint Research Projects//Japanese Society for the Promotion of Science/ ; Core-to-Core program (Asia-Africa Science Platforms)//Japanese Society for the Promotion of Science/ ; Y29-1-8//Leading Academia in Marine, Environmental Pollution Research/ ; }, mesh = {Aedes/*microbiology ; Animals ; DNA, Ribosomal ; Female ; Male ; Mosquito Vectors/*microbiology ; Philippines ; Phylogeny ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Recent reports reveal the presence of Wolbachia in Ae. aegypti. Our study presents additional support for Wolbachia infection in Ae. aegypti by screening field-collected adult mosquitoes using two Wolbachia-specific molecular makers.

METHODS: A total of 672 Ae. aegypti adult mosquitoes were collected from May 2014 to January 2015 in Metropolitan Manila. Each individual sample was processed and screened for the presence of Wolbachia by selected markers, Wolbachia-specific 16S rDNA and its surface protein (wsp), under optimized PCR conditions and sequenced.

RESULTS: Totals of 113 (16.8%) and 89 (13.2%) individual mosquito samples were determined to be infected with Wolbachia using the wsp and 16S rDNA markers, respectively. The Ae. aegpyti wsp sample sequences were similar or identical to five known Wolbachia strains belonging to supergroups A and B while the majority of 16S rDNA sample sequences were similar to strains belonging to supergroup B. Overall, 80 (11.90%) individual mosquito samples showed positive amplifications in both markers and 69% showed congruence in supergroup identification (supergroup B).

CONCLUSIONS: By utilizing two Wolbachia-specific molecular makers, our study demonstrated the presence of Wolbachia from individual Ae. aegypti samples. Our results showed a low Wolbachia infection rate and inferred the detected strains belong to either supergroups A and B.}, } @article {pmid31340757, year = {2019}, author = {He, Z and Zheng, Y and Yu, WJ and Fang, Y and Mao, B and Wang, YF}, title = {How do Wolbachia modify the Drosophila ovary? New evidences support the "titration-restitution" model for the mechanisms of Wolbachia-induced CI.}, journal = {BMC genomics}, volume = {20}, number = {1}, pages = {608}, pmid = {31340757}, issn = {1471-2164}, support = {31672352//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Cytosol ; Drosophila melanogaster/*microbiology ; Female ; Gene Expression Regulation, Developmental ; Genes, Insect ; Male ; MicroRNAs/genetics ; Ovary/*microbiology ; RNA, Messenger/genetics ; RNA-Seq ; Testis/microbiology ; Transcriptome ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Cytoplasmic incompatibility (CI) is the most common phenotype induced by endosymbiont Wolbachia and results in embryonic lethality when Wolbachia-modified sperm fertilize eggs without Wolbachia. However, eggs carrying the same strain of Wolbachia can rescue this embryonic death, thus producing viable Wolbachia-infected offspring. Hence Wolbachia can be transmitted mainly by hosts' eggs. One of the models explaining CI is "titration-restitution", which hypothesized that Wolbachia titrated-out some factors from the sperm and the Wolbachia in the egg would restitute the factors after fertilization. However, how infected eggs rescue CI and how hosts' eggs ensure the proliferation and transmission of Wolbachia are not well understood.

RESULTS: By RNA-seq analyses, we first compared the transcription profiles of Drosophila melanogaster adult ovaries with and without the wMel Wolbachia and identified 149 differentially expressed genes (DEGs), of which 116 genes were upregulated and 33 were downregulated by Wolbachia infection. To confirm the results obtained from RNA-seq and to screen genes potentially associated with reproduction, 15 DEGs were selected for quantitative RT-PCR (qRT-PCR). Thirteen genes showed the same changing trend as RNA-seq analyses. To test whether these genes are associated with CI, we also detected their expression levels in testes. Nine of them exhibited different changing trends in testes from those in ovaries. To investigate how these DEGs were regulated, sRNA sequencing was performed and identified seven microRNAs (miRNAs) that were all upregulated in fly ovaries by Wolbachia infection. Matching of miRNA and mRNA data showed that these seven miRNAs regulated 15 DEGs. Wolbachia-responsive genes in fly ovaries were involved in biological processes including metabolism, transportation, oxidation-reduction, immunity, and development.

CONCLUSIONS: Comparisons of mRNA and miRNA data from fly ovaries revealed 149 mRNAs and seven miRNAs that exhibit significant changes in expression due to Wolbachia infection. Notably, most of the DEGs showed variation in opposite directions in ovaries versus testes in the presence of Wolbachia, which generally supports the "titration-restitution" model for CI. Furthermore, genes related to metabolism were upregulated, which may benefit maximum proliferation and transmission of Wolbachia. This provides new insights into the molecular mechanisms of Wolbachia-induced CI and Wolbachia dependence on host ovaries.}, } @article {pmid31340171, year = {2019}, author = {Pogorevc, D and Panter, F and Schillinger, C and Jansen, R and Wenzel, SC and Müller, R}, title = {Production optimization and biosynthesis revision of corallopyronin A, a potent anti-filarial antibiotic.}, journal = {Metabolic engineering}, volume = {55}, number = {}, pages = {201-211}, doi = {10.1016/j.ymben.2019.07.010}, pmid = {31340171}, issn = {1096-7184}, mesh = {Anti-Bacterial Agents/*biosynthesis ; Filaricides/*metabolism ; Lactones/*metabolism ; *Microorganisms, Genetically-Modified/genetics/metabolism ; *Myxococcus xanthus/genetics/metabolism ; }, abstract = {Corallopyronins (COR) are α-pyrone antibiotics from myxobacteria representing highly promising lead structures for the development of antibacterial therapeutic agents. Their ability to inhibit RNA polymerase through interaction with the "switch region", a novel target, distant from binding sites of previously characterized RNA polymerase inhibitors (e.g. rifampicin), makes them particularly promising as antibiotic candidates. Corallopyronin A is currently also investigated as a lead compound for the treatment of lymphatic filariasis because of its superb activity against the nematode symbiont Wolbachia. As total synthesis is not a valid production option biotechnological optimization of compound supply is of utmost importance to further develop this highly potent compound class. Here we describe decisive improvements of the previously reported heterologous COR production and engineering platform yielding production of ~100 mg/L COR A. Furthermore, we provide a revised model of COR biosynthesis shedding light on the function of several biosynthetic proteins, including an unusual ECH-like enzyme providing dehydration functionality in trans and an uncharacterized protein conferring COR self-resistance in the myxobacterial heterologous host Myxococcus xanthus DK1622. We also report two new COR derivatives, COR D and oxyCOR A discovered in genetically engineered strains.}, } @article {pmid31334893, year = {2019}, author = {Treanor, D and Hughes, WOH}, title = {Limited female dispersal predicts the incidence of Wolbachia across ants (Hymenoptera: Formicidae).}, journal = {Journal of evolutionary biology}, volume = {32}, number = {10}, pages = {1163-1170}, doi = {10.1111/jeb.13510}, pmid = {31334893}, issn = {1420-9101}, mesh = {Animals ; Ants/genetics/*microbiology ; Female ; Species Specificity ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia is perhaps the greatest panzootic in the history of life on Earth, yet remarkably little is known regarding the factors that determine its incidence across species. One possibility is that Wolbachia more easily invades species with structured populations, due to the increased strength of genetic drift and higher initial frequency of infection. This should enable strains that induce mating incompatibilities to more easily cross the threshold prevalence above which they spread to either fixation or a stable equilibrium infection prevalence. Here, we provide empirical support for this hypothesis by analysing the relationship between female dispersal (as a proxy for population structure) and the incidence of Wolbachia across 250 species of ants. We show that species in which the dispersal of reproductive females is limited are significantly more likely to be infected with Wolbachia than species whose reproductive ecology is consistent with significant dispersal of females, and that this relationship remains after controlling for host phylogeny. We suggest that structured host populations, in this case resulting from limited female dispersal, may be an important feature determining how easily Wolbachia becomes successfully established in a novel host, and thus its occurrence across a wide diversity of invertebrate hosts.}, } @article {pmid31334887, year = {2019}, author = {Barash, NR and Thomas, B and Birkenheuer, AJ and Breitschwerdt, EB and Lemler, E and Qurollo, BA}, title = {Prevalence of Babesia spp. and clinical characteristics of Babesia vulpes infections in North American dogs.}, journal = {Journal of veterinary internal medicine}, volume = {33}, number = {5}, pages = {2075-2081}, pmid = {31334887}, issn = {1939-1676}, mesh = {Animals ; Babesia/classification/*isolation & purification ; Babesiosis/*epidemiology/transmission ; Coinfection/veterinary ; Dog Diseases/epidemiology/microbiology/*parasitology/pathology ; Dogs ; North America/epidemiology ; Polymerase Chain Reaction/veterinary ; Prevalence ; }, abstract = {BACKGROUND: Babesiosis is an important cause of thrombocytopenia and hemolytic anemia in dogs. Babesia vulpes, reported in European dogs and North American foxes, rarely has been reported in domestic North American dogs. Newly optimized polymerase chain reaction (PCR) primers facilitate more sensitive amplification of B. vulpes DNA.

OBJECTIVES: To determine the prevalence of Babesia sp. infections in dogs being tested for Babesia infection, and to describe co-infections and clinicopathologic abnormalities in B. vulpes positive dogs.

ANIMALS: Dog blood or tissue samples (n = 9367) submitted to a diagnostic laboratory between June 2015 and June 2018 were tested using an optimized Babesia PCR assay.

METHODS: Comprehensive canine vector-borne disease diagnostic testing was performed on convenience samples.

RESULTS: Babesia sp. DNA was amplified from 269/9367 (2.9%) North American dogs. Babesia sp. infections included B. gibsoni monoinfection (157; 1.7%), B. vulpes monoinfection (19; 0.20%), and B. gibsoni and B. vulpes coinfection (29; 0.31%). Forty-three of the 48 total B. vulpes-infected dogs were American Pit Bull Terrier-type breeds, of which 36 historically were involved with dog fights. Coinfections with Mycoplasma, Dirofilaria immitis, or Wolbachia and coexposures to Bartonella, Ehrlichia, and Rickettsia spp. were documented in B. vulpes-infected dogs. Clinicopathologic data in B. vulpes-infected dogs both with and without coinfections included anemia, thrombocytopenia, hyperglobulinemia, hypoalbuminemia, and proteinuria.

Babesia vulpes infection in domestic North American dogs is commonly found in conjunction with other coinfections, including B. gibsoni and hemotropic Mycoplasma. Similar to B. gibsoni, dog-to-dog transmission of B. vulpes may be a frequent mode of transmission.}, } @article {pmid31323841, year = {2019}, author = {Bakowski, MA and McNamara, CW}, title = {Advances in Antiwolbachial Drug Discovery for Treatment of Parasitic Filarial Worm Infections.}, journal = {Tropical medicine and infectious disease}, volume = {4}, number = {3}, pages = {}, pmid = {31323841}, issn = {2414-6366}, abstract = {The intracellular bacteria now known as Wolbachia were first described in filarial worms in the 1970s, but the idea of Wolbachia being used as a macrofilaricidal target did not gain wide attention until the early 2000s, with research in filariae suggesting the requirement of worms for the endosymbiont. This new-found interest prompted the eventual organization of the Anti-Wolbachia Consortium (A-WOL) at the Liverpool School of Tropical Medicine, who, among others have been active in the field of antiwolbachial drug discovery to treat filarial infections. Clinical proof of concept studies using doxycycline demonstrated the utility of the antiwolbachial therapy, but efficacious treatments were of long duration and not safe for all infected. With the advance of robotics, automation, and high-speed computing, the search for superior antiwolbachials shifted away from smaller studies with a select number of antibiotics to high-throughput screening approaches, centered largely around cell-based phenotypic screens due to the rather limited knowledge about, and tools available to manipulate, this bacterium. A concomitant effort was put towards developing validation approaches and in vivo models supporting drug discovery efforts. In this review, we summarize the strategies behind and outcomes of recent large phenotypic screens published within the last 5 years, hit compound validation approaches and promising candidates with profiles superior to doxycycline, including ones positioned to advance into clinical trials for treatment of filarial worm infections.}, } @article {pmid31323076, year = {2019}, author = {Cheng, D and Chen, S and Huang, Y and Pierce, NE and Riegler, M and Yang, F and Zeng, L and Lu, Y and Liang, G and Xu, Y}, title = {Symbiotic microbiota may reflect host adaptation by resident to invasive ant species.}, journal = {PLoS pathogens}, volume = {15}, number = {7}, pages = {e1007942}, pmid = {31323076}, issn = {1553-7374}, mesh = {Adaptation, Physiological ; Animals ; Ants/*microbiology/*physiology ; Diet ; Ecosystem ; Feeding Behavior ; Host Microbial Interactions/*physiology ; *Introduced Species ; Microbiota/*physiology ; Species Specificity ; Symbiosis/physiology ; Vitamin B Complex/metabolism ; Wolbachia/physiology ; }, abstract = {Exotic invasive species can influence the behavior and ecology of native and resident species, but these changes are often overlooked. Here we hypothesize that the ghost ant, Tapinoma melanocephalum, living in areas that have been invaded by the red imported fire ant, Solenopsis invicta, displays behavioral differences to interspecific competition that are reflected in both its trophic position and symbiotic microbiota. We demonstrate that T. melanocephalum workers from S. invicta invaded areas are less aggressive towards workers of S. invicta than those inhabiting non-invaded areas. Nitrogen isotope analyses reveal that colonies of T. melanocephalum have protein-rich diets in S. invicta invaded areas compared with the carbohydrate-rich diets of colonies living in non-invaded areas. Analysis of microbiota isolated from gut tissue shows that T. melanocephalum workers from S. invicta invaded areas also have different bacterial communities, including a higher abundance of Wolbachia that may play a role in vitamin B provisioning. In contrast, the microbiota of workers of T. melanocephalum from S. invicta-free areas are dominated by bacteria from the orders Bacillales, Lactobacillales and Enterobacteriales that may be involved in sugar metabolism. We further demonstrate experimentally that the composition and structure of the bacterial symbiont communities as well as the prevalence of vitamin B in T. melanocephalum workers from S. invicta invaded and non-invaded areas can be altered if T. melanocephalum workers are supplied with either protein-rich or carbohydrate-rich food. Our results support the hypothesis that bacterial symbiont communities can help hosts by buffering behavioral changes caused by interspecies competition as a consequence of biological invasions.}, } @article {pmid31316207, year = {2019}, author = {Zheng, X and Zhang, D and Li, Y and Yang, C and Wu, Y and Liang, X and Liang, Y and Pan, X and Hu, L and Sun, Q and Wang, X and Wei, Y and Zhu, J and Qian, W and Yan, Z and Parker, AG and Gilles, JRL and Bourtzis, K and Bouyer, J and Tang, M and Zheng, B and Yu, J and Liu, J and Zhuang, J and Hu, Z and Zhang, M and Gong, JT and Hong, XY and Zhang, Z and Lin, L and Liu, Q and Hu, Z and Wu, Z and Baton, LA and Hoffmann, AA and Xi, Z}, title = {Incompatible and sterile insect techniques combined eliminate mosquitoes.}, journal = {Nature}, volume = {572}, number = {7767}, pages = {56-61}, pmid = {31316207}, issn = {1476-4687}, mesh = {Aedes/growth & development/*microbiology/*physiology ; Animals ; China ; Copulation ; Feasibility Studies ; Female ; Humans ; Insect Bites and Stings/prevention & control ; Larva/growth & development/microbiology/physiology ; Male ; Mosquito Control/*methods ; Mosquito Vectors/growth & development/*microbiology/*physiology ; Quality Control ; Reproduction ; Wolbachia/*pathogenicity ; }, abstract = {The radiation-based sterile insect technique (SIT) has successfully suppressed field populations of several insect pest species, but its effect on mosquito vector control has been limited. The related incompatible insect technique (IIT)-which uses sterilization caused by the maternally inherited endosymbiotic bacteria Wolbachia-is a promising alternative, but can be undermined by accidental release of females infected with the same Wolbachia strain as the released males. Here we show that combining incompatible and sterile insect techniques (IIT-SIT) enables near elimination of field populations of the world's most invasive mosquito species, Aedes albopictus. Millions of factory-reared adult males with an artificial triple-Wolbachia infection were released, with prior pupal irradiation of the released mosquitoes to prevent unintentionally released triply infected females from successfully reproducing in the field. This successful field trial demonstrates the feasibility of area-wide application of combined IIT-SIT for mosquito vector control.}, } @article {pmid31312785, year = {2019}, author = {Dietrich, CF and Chaubal, N and Hoerauf, A and Kling, K and Piontek, MS and Steffgen, L and Mand, S and Dong, Y}, title = {Review of Dancing Parasites in Lymphatic Filariasis.}, journal = {Ultrasound international open}, volume = {5}, number = {2}, pages = {E65-E74}, pmid = {31312785}, issn = {2509-596X}, abstract = {Lymphatic filariasis is an infection transmitted by blood-sucking mosquitoes with filarial nematodes of the species Wuchereria bancrofti, Brugia malayi und B. timori . It is prevalent in tropical countries throughout the world, with more than 60 million people infected and more than 1 billion living in areas with the risk of transmission. Worm larvae with a length of less than 1 mm are transmitted by mosquitoes, develop in human lymphatic tissue to adult worms with a length of 7-10 cm, live in the human body for up to 10 years and produce millions of microfilariae, which can be transmitted further by mosquitoes. The adult worms can be easily observed by ultrasonography because of their size and fast movements (the so-called "filarial dance sign"), which can be differentiated from other movements (e. g., blood in venous vessels) by their characteristic movement profile in pulsed-wave Doppler mode. Therapeutic options include (combinations of) ivermectin, albendazole, diethylcarbamazine and doxycycline. The latter depletes endosymbiotic Wolbachia bacteria from the worms and thus sterilizes and later kills the adult worms (macrofilaricidal or adulticidal effect).}, } @article {pmid31311998, year = {2019}, author = {Ke, F and You, S and Huang, S and Chen, W and Liu, T and He, W and Xie, D and Li, Q and Lin, X and Vasseur, L and Gurr, GM and You, M}, title = {Herbivore range expansion triggers adaptation in a subsequently-associated third trophic level species and shared microbial symbionts.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {10314}, pmid = {31311998}, issn = {2045-2322}, mesh = {Adaptation, Physiological ; Animals ; Brassica/*parasitology ; China ; DNA, Mitochondrial ; Genetic Variation ; Herbivory/*physiology ; Hymenoptera/*classification/physiology ; Introduced Species ; Moths/*classification/physiology ; Phylogeography ; Symbiosis ; }, abstract = {Invasive species may change the life history strategies, distribution, genetic configuration and trophic interactions of native species. The diamondback moth, Plutella xylostella L., is an invasive herbivore attacking cultivated and wild brassica plants worldwide. Here we present phylogeographic analyses of P. xylostella and one of its major parasitoids, Cotesia vestalis, using mitochondrial markers, revealing the genetic diversity and evolutionary history of these two species. We find evidence that C. vestalis originated in Southwest China, then adapted to P. xylostella as a new host by ecological sorting as P. xylostella expanded its geographic range into this region. Associated with the expansion of P. xylostella, Wolbachia symbionts were introduced into local populations of the parasitoid through horizontal transfer from its newly associated host. Insights into the evolutionary history and phylogeographic system of the herbivore and its parasitoid provide an important basis for better understanding the impacts of biological invasion on genetic configuration of local species.}, } @article {pmid31302148, year = {2020}, author = {Iftikhar, M and Iftikhar, S and Sohail, A and Javed, S}, title = {AI- modelling of molecular identification and feminization of wolbachia infected Aedes aegypti.}, journal = {Progress in biophysics and molecular biology}, volume = {150}, number = {}, pages = {104-111}, doi = {10.1016/j.pbiomolbio.2019.07.001}, pmid = {31302148}, issn = {1873-1732}, mesh = {Aedes/*genetics/microbiology ; Animals ; *Artificial Intelligence ; Computer Simulation ; Feminization/*genetics ; Humans ; Infections ; Male ; Models, Biological ; Models, Theoretical ; Mosquito Vectors/*microbiology ; Reproduction/genetics ; Systems Analysis ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: The genetic control strategies of vector borne diseases includes the replacement of a vector population by "disease-refractory" mosquitoes and the release of mosquitoes with a gene to control the vector's reproduction rates. Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. Wolbachia infected male mosquitos have been used in different experimental trials around the world to suppress the target population of Aedes aegypti and this genetic control strategy has proved to be a promising alternative to other treatment strategies. Due to certain limitations, the successful application of this strategy is still awaited.

METHODS: Mathematical frame work for Wolbachia induced genetic control strategy has been developed in this article. With the aid of Artificial Intelligence (AI) tools, accurate parametric values are depicted. For the first time, the model is well synchronized with the experimental findings. The model is comprised of the generalized varying coefficient and multiple mating rates between infected and uninfected compartments of Aedes aegypti dengue to forecast the disease control.

RESULTS: Two mathematical models are developed in this article to demonstrate different mating rates of the genetic control strategy. The important parameters and time varying coefficients are well demonstrated with the aid of numerical computations. The resulting thresholds and forecasting may prove to be a useful tool for future experimental studies.

CONCLUSIONS: From our analysis, we have concluded that the genetic control strategy is a promising technique and the role of Wolbachia infected male mosquitos, in genetic control strategies, can be better interpreted in an inexpensive manner with the aid of a theoretical model.}, } @article {pmid31300838, year = {2020}, author = {White, JA and Styer, A and Rosenwald, LC and Curry, MM and Welch, KD and Athey, KJ and Chapman, EG}, title = {Endosymbiotic Bacteria Are Prevalent and Diverse in Agricultural Spiders.}, journal = {Microbial ecology}, volume = {79}, number = {2}, pages = {472-481}, pmid = {31300838}, issn = {1432-184X}, support = {0224651//U.S. Department of Agriculture/ ; 148-502-16-377//Kentucky Science and Engineering Foundation/ ; 148-502-10-261//Kentucky Science and Engineering Foundation/ ; }, mesh = {Animals ; Bacteria/classification/*isolation & purification ; Bacterial Physiological Phenomena ; Bacteriological Techniques/*methods ; Entomology/*methods ; Food Deprivation ; High-Throughput Nucleotide Sequencing ; Kentucky ; *Microbiota/genetics ; Polymerase Chain Reaction ; Spiders/*microbiology ; Symbiosis/*physiology ; }, abstract = {Maternally inherited bacterial endosymbionts are common in arthropods, but their distribution and prevalence are poorly characterized in many host taxa. Initial surveys have suggested that vertically transmitted symbionts may be particularly common in spiders (Araneae). Here, we used diagnostic PCR and high-throughput sequencing to evaluate symbiont infection in 267 individual spiders representing 14 species (3 families) of agricultural spiders. We found 27 operational taxonomic units (OTUs) that are likely endosymbiotic, including multiple strains of Wolbachia, Rickettsia, and Cardinium, which are all vertically transmitted and frequently associated with reproductive manipulation of arthropod hosts. Additional strains included Rickettsiella, Spiroplasma, Rhabdochlamydia, and a novel Rickettsiales, all of which could range from pathogenic to mutualistic in their effects upon their hosts. Seventy percent of spider species had individuals that tested positive for one or more endosymbiotic OTUs, and specimens frequently contained multiple symbiotic strain types. The most symbiont-rich species, Idionella rugosa, had eight endosymbiotic OTUs, with as many as five present in the same specimen. Individual specimens within infected spider species had a variety of symbiotypes, differing from one another in the presence or absence of symbiotic strains. Our sample included both starved and unstarved specimens, and dominant bacterial OTUs were consistent per host species, regardless of feeding status. We conclude that spiders contain a remarkably diverse symbiotic microbiota. Spiders would be an informative group for investigating endosymbiont population dynamics in time and space, and unstarved specimens collected for other purposes (e.g., food web studies) could be used, with caution, for such investigations.}, } @article {pmid31300646, year = {2019}, author = {Reveillaud, J and Bordenstein, SR and Cruaud, C and Shaiber, A and Esen, ÖC and Weill, M and Makoundou, P and Lolans, K and Watson, AR and Rakotoarivony, I and Bordenstein, SR and Eren, AM}, title = {Author Correction: The Wolbachia mobilome in Culex pipiens includes a putative plasmid.}, journal = {Nature communications}, volume = {10}, number = {1}, pages = {3153}, doi = {10.1038/s41467-019-11234-5}, pmid = {31300646}, issn = {2041-1723}, support = {T32 EB009412/EB/NIBIB NIH HHS/United States ; }, abstract = {An amendment to this paper has been published and can be accessed via a link at the top of the paper.}, } @article {pmid31300097, year = {2019}, author = {Boucheikhchoukh, M and Mechouk, N and Benakhla, A and Raoult, D and Parola, P}, title = {Molecular evidence of bacteria in Melophagus ovinus sheep keds and Hippobosca equina forest flies collected from sheep and horses in northeastern Algeria.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {65}, number = {}, pages = {103-109}, doi = {10.1016/j.cimid.2019.05.010}, pmid = {31300097}, issn = {1878-1667}, mesh = {Algeria/epidemiology ; Anaplasmataceae/genetics ; Animals ; Bacteria/*genetics/isolation & purification ; Bartonella/genetics ; DNA, Bacterial/*genetics ; Diptera/*microbiology ; Disease Vectors ; Female ; Forests ; Horses/*parasitology ; Male ; Rickettsia/genetics ; Sheep/*parasitology ; }, abstract = {The sheep ked, Melophagus ovinus, and the forest fly, Hippobosca equina, are parasitic dipteran insects of veterinary importance. As hematophagous insects, they might be considered as potential vectors of diseases which may be transmissible to humans and animals. The purpose of this study was to present initial primary data about these two species in Algeria. To do so, we conducted a molecular survey to detect the presence of bacterial DNA in flies collected in Algeria. A total of 712 flies including, 683 Melophagus ovinus and 29 Hippobosca equina were collected from two regions in northeastern Algeria. Monitoring the monthly kinetics of M. ovinus infestations showed something resembling annual activity, with a high prevalence in January (21.67%) and May (20.94%). Real-time quantitative PCR assays showed that for 311 tested flies, 126 were positive for the Bartonella spp. rRNA intergenic spacer gene and 77 were positive for Anaplasmataceae. A random selection of positive samples was submitted for sequencing. The DNA of Bartonella chomelii and Bartonella melophagi were amplified in, respectively, five and four H. equina. 25 M. ovinus positive samples were infected by Bartonella melophagi. Amplification and sequencing of the Anaplasma spp. 23S rRNA gene revealed that both species were infected by Wolbachia sp. which had previously been detected in Cimex lectularius bed bugs. Overall, this study expanded knowledge about bacteria present in parasitic flies of domestic animals in Algeria.}, } @article {pmid31279737, year = {2019}, author = {Nebbak, A and Dahmana, H and Almeras, L and Raoult, D and Boulanger, N and Jaulhac, B and Mediannikov, O and Parola, P}, title = {Co-infection of bacteria and protozoan parasites in Ixodes ricinus nymphs collected in the Alsace region, France.}, journal = {Ticks and tick-borne diseases}, volume = {10}, number = {6}, pages = {101241}, doi = {10.1016/j.ttbdis.2019.06.001}, pmid = {31279737}, issn = {1877-9603}, mesh = {Animals ; Babesia/classification/isolation & purification ; Bartonella/classification/isolation & purification ; Borrelia/isolation & purification ; France ; Ixodes/growth & development/*microbiology/*parasitology ; Nymph/growth & development/microbiology/parasitology ; Phylogeny ; Polymerase Chain Reaction ; Rickettsia/isolation & purification ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Wolbachia/isolation & purification ; }, abstract = {Fifty nymphal Ixodes ricinus ticks collected in Alsace, France, identified by morphological criteria and using MALDI-TOF MS, were tested by PCR to detect tick-associated bacteria and protozoan parasites. Seventy percent (35/50) of ticks contained at least one microorganism; 26% (9/35) contained two or more species. Several human pathogens were identified including Borrelia burgdorferi s.s. (4%), Borrelia afzelii (2%), Borrelia garinii (2%), Borrelia valaisiana (4%), Borrelia miyamotoi (2%), Rickettsia helvetica (6%) and "Babesia venatorum" (2%). Bartonella spp. (10%) and a Wolbachia spp. (8%) were also detected. The most common co-infections involved Anaplasmataceae with Borrelia spp. (4%), Anaplasmataceae with Bartonella spp. (6%) and Anaplasmataceae with Rickettsia spp. (6%). Co-infection involving three different groups of bacteria was seen between bacteria of the family Anaplasmataceae, Borrelia spp. and Bartonella spp. (2%). Results highlight the panel of infectious agents carried by Ixodes ricinus. Co-infection suggests the possibility of transmission of more than one pathogen to human and animals during tick blood feeding.}, } @article {pmid31274094, year = {2019}, author = {Naciri, M}, title = {[Wolbachia bacteria inhibits mosquito infection by various human pathogens].}, journal = {Medecine sciences : M/S}, volume = {35}, number = {6-7}, pages = {584-585}, doi = {10.1051/medsci/2019115}, pmid = {31274094}, issn = {1958-5381}, mesh = {Aedes/microbiology/virology ; Animals ; Antibiosis ; Culicidae/*microbiology/virology ; Dengue Virus ; Humans ; Longevity ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology/virology ; Wolbachia/*physiology ; }, } @article {pmid31267762, year = {2019}, author = {Lima-Barbero, JF and Díaz-Sanchez, S and Sparagano, O and Finn, RD and de la Fuente, J and Villar, M}, title = {Metaproteomics characterization of the alphaproteobacteria microbiome in different developmental and feeding stages of the poultry red mite Dermanyssus gallinae (De Geer, 1778).}, journal = {Avian pathology : journal of the W.V.P.A}, volume = {48}, number = {sup1}, pages = {S52-S59}, doi = {10.1080/03079457.2019.1635679}, pmid = {31267762}, issn = {1465-3338}, mesh = {Alphaproteobacteria/*isolation & purification ; Animals ; Chickens/*parasitology ; Female ; *Microbiota ; Mites/*microbiology ; Poultry Diseases/*parasitology ; Proteomics ; }, abstract = {The poultry red mite (PRM), Dermanyssus gallinae (De Geer, 1778), is a worldwide distributed ectoparasite and considered a major pest affecting the laying hen industry in Europe. Based on available information in other ectoparasites, the mite microbiome might participate in several biological processes and the acquisition, maintenance and transmission of pathogens. However, little is known about the role of PRM as a mechanical carrier or a biological vector in the transmission of pathogenic bacteria. Herein, we used a metaproteomics approach to characterize the alphaproteobacteria in the microbiota of PRM, and variations in its profile with ectoparasite development (nymphs vs. adults) and feeding (unfed vs. fed). The results showed that the bacterial community associated with D. gallinae was mainly composed of environmental and commensal bacteria. Putative symbiotic bacteria of the genera Wolbachia, C. Tokpelaia and Sphingomonas were identified, together with potential pathogenic bacteria of the genera Inquilinus, Neorickettsia and Roseomonas. Significant differences in the composition of alphaproteobacterial microbiota were associated with mite development and feeding, suggesting that bacteria have functional implications in metabolic pathways associated with blood feeding. These results support the use of metaproteomics for the characterization of alphaproteobacteria associated with the D. gallinae microbiota that could provide relevant information for the understanding of mite-host interactions and the development of potential control interventions. Research highlights Metaproteomics is a valid approach for microbiome characterization in ectoparasites. Alphaproteobacteria putative bacterial symbionts were identified in D. gallinae. Mite development and feeding were related to variations in bacterial community. Potentially pathogenic bacteria were identified in mite microbiota.}, } @article {pmid31265751, year = {2020}, author = {Yang, K and Xie, K and Zhu, YX and Huo, SM and Hoffmann, A and Hong, XY}, title = {Wolbachia dominate Spiroplasma in the co-infected spider mite Tetranychus truncatus.}, journal = {Insect molecular biology}, volume = {29}, number = {1}, pages = {19-37}, doi = {10.1111/imb.12607}, pmid = {31265751}, issn = {1365-2583}, mesh = {Animals ; Coinfection/microbiology ; Female ; Fertility ; Gene Expression Profiling ; Male ; Spiroplasma/*physiology ; Symbiosis ; Tetranychidae/genetics/metabolism/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia and Spiroplasma are both maternally inherited endosymbionts in arthropods, and they can co-infect the same species. However, how they interact with each other in the same host is not clear. Here we investigate a co-infected Tetranychus truncatus spider mite strain that shares the same genetic background with singly infected and uninfected strains to detect the impacts of the two symbionts on their host. We found that Wolbachia-infected and Spiroplasma-infected mites can suffer significant fitness costs involving decreased fecundity, although with no effect on lifespan or development. Wolbachia induced incomplete cytoplasmic incompatibility in T. truncatus both in singly infected and doubly infected strains, resulting in female killing. In both females and males of the co-infected spider mite strain, Wolbachia density was higher than Spiroplasma density. Transcriptome analysis of female adults showed that the most differentially expressed genes were found between the co-infected strain and both the singly infected Spiroplasma strain and uninfected strain. The Wolbachia strain had the fewest differentially expressed genes compared with the co-infected strain, consistent with the higher density of Wolbachia in the co-infected strain. Wolbachia, therefore, appears to have a competitive advantage in host mites over Spiroplasma and is likely maintained in populations by cytoplasmic incompatibility despite having deleterious fitness effects.}, } @article {pmid31259313, year = {2019}, author = {Rocha, MN and Duarte, MM and Mansur, SB and Silva, BDME and Pereira, TN and Adelino, TÉR and Giovanetti, M and Alcantara, LCJ and Santos, FM and Costa, VRM and Teixeira, MM and Iani, FCM and Costa, VV and Moreira, LA}, title = {Pluripotency of Wolbachia against Arboviruses: the case of yellow fever.}, journal = {Gates open research}, volume = {3}, number = {}, pages = {161}, pmid = {31259313}, issn = {2572-4754}, abstract = {Background: Yellow fever outbreaks have re-emerged in Brazil during 2016-18, with mortality rates up to 30%. Although urban transmission has not been reported since 1942, the risk of re-urbanization of yellow fever is significant, as Aedes aegypti is present in most tropical and sub-tropical cities in the World and still remains the main vector of urban YFV. Although the YFV vaccine is safe and effective, it does not always reach populations at greatest risk of infection and there is an acknowledged global shortage of vaccine supply. The introgression of Wolbachia bacteria into Ae. aegypti mosquito populations is being trialed in several countries (www.worldmosquito.org) as a biocontrol method against dengue, Zika and chikungunya. Here, we studied the ability of Wolbachia to reduce the transmission potential of Ae. aegypti mosquitoes for Yellow fever virus (YFV). Methods: Two recently isolated YFV (primate and human) were used to challenge field-derived wild-type and Wolbachia-infected (wMel +) Ae. aegypti mosquitoes. The YFV infection status was followed for 7, 14 and 21 days post-oral feeding (dpf). The YFV transmission potential of mosquitoes was evaluated via nano-injection of saliva into uninfected mosquitoes or by inoculation in mice. Results: We found that Wolbachia was able to significantly reduce the prevalence of mosquitoes with YFV infected heads and thoraces for both viral isolates. Furthermore, analyses of mosquito saliva, through indirect injection into naïve mosquitoes or via interferon-deficient mouse model, indicated Wolbachia was associated with profound reduction in the YFV transmission potential of mosquitoes (14dpf). Conclusions: Our results suggest that Wolbachia introgression could be used as a complementary strategy for prevention of urban yellow fever transmission, along with the human vaccination program.}, } @article {pmid31254009, year = {2019}, author = {Kelly, M and Price, SL and de Oliveira Ramalho, M and Moreau, CS}, title = {Diversity of Wolbachia Associated with the Giant Turtle Ant, Cephalotes atratus.}, journal = {Current microbiology}, volume = {76}, number = {11}, pages = {1330-1337}, pmid = {31254009}, issn = {1432-0991}, support = {1559779//NSF DBI/ ; 1900357//National Science Foundation/ ; }, mesh = {Animals ; Ants/*microbiology/physiology ; *Biodiversity ; Multilocus Sequence Typing ; Phylogeny ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Symbiotic relationships between organisms are common throughout the tree of life, and often these organisms share an evolutionary history. In turtle ants (Cephalotes), symbiotic associations with bacteria are known to be especially important for supplementing the nutrients that their herbivorous diets do not provide. However, much remains unknown about the diversity of many common bacterial symbionts with turtle ants, such as Wolbachia. Here, we surveyed the diversity of Wolbachia, focusing on one species of turtle ant with a particularly wide geographic range, Cephalotes atratus. Colonies were collected from the entire range of C. atratus, and we detected the presence of Wolbachia by sequencing multiple individuals per colony for wsp. Then, using the multilocus sequence typing (MLST) approach, we determined each individual's unique sequence type (ST) based on comparison to sequences published in the Wolbachia MLST Database (https://pubmlst.org/wolbachia/). The results of this study suggest that there is a high level of diversity of Wolbachia strains among colonies from different regions, while the diversity within colonies is very low. Additionally, 13 novel variants (alleles) were uncovered. These results suggest that the level of diversity of Wolbachia within species is affected by geography, and the high level of diversity observed among Cephalotes atratus populations may be explained by their wide geographic range.}, } @article {pmid31247412, year = {2019}, author = {Sicard, M and Bonneau, M and Weill, M}, title = {Wolbachia prevalence, diversity, and ability to induce cytoplasmic incompatibility in mosquitoes.}, journal = {Current opinion in insect science}, volume = {34}, number = {}, pages = {12-20}, doi = {10.1016/j.cois.2019.02.005}, pmid = {31247412}, issn = {2214-5753}, mesh = {Animals ; Culicidae/*microbiology ; Female ; Genes, Bacterial ; Male ; *Mosquito Control ; *Wolbachia ; }, abstract = {To protect humans and domestic animals from mosquito borne diseases, alternative methods to chemical insecticides have to be found. Pilot studies using the vertically transmitted bacterial endosymbiont Wolbachia were already launched in different parts of the world. Wolbachia can be used either in Incompatible Insect Technique (IIT), to decrease mosquito population, or to decrease the ability of mosquitoes to transmit pathogens. Not all mosquito species are naturally infected with Wolbachia: while in Culex pipiens and Aedes albopictus almost all individuals harbor Wolbachia, putative infections have to be further investigated in Anopheles species and in Aedes aegypti. All Wolbachia-based control methods rely on the ability of Wolbachia to induce cytoplasmic incompatibility (CI) resulting in embryonic death in incompatible crossings. Knowledge on CI diversity in mosquito is required to find the better Wolbachia-mosquito associations to optimize the success of both 'sterile insect' and 'pathogen blocking' Wolbachia-based methods.}, } @article {pmid31242186, year = {2019}, author = {Shropshire, JD and Bordenstein, SR}, title = {Two-By-One model of cytoplasmic incompatibility: Synthetic recapitulation by transgenic expression of cifA and cifB in Drosophila.}, journal = {PLoS genetics}, volume = {15}, number = {6}, pages = {e1008221}, pmid = {31242186}, issn = {1553-7404}, support = {P30 DK058404/DK/NIDDK NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified/genetics/growth & development ; Bacteriophages/*genetics ; Cytoplasm/genetics/metabolism/microbiology ; Disease Vectors ; Drosophila melanogaster/*genetics/growth & development/microbiology ; Female ; Gene Expression Regulation/genetics ; Male ; Maternal Inheritance/genetics ; Reproduction/genetics ; Viral Proteins/*genetics ; Wolbachia/*genetics/pathogenicity/virology ; }, abstract = {Wolbachia are maternally inherited bacteria that infect arthropod species worldwide and are deployed in vector control to curb arboviral spread using cytoplasmic incompatibility (CI). CI kills embryos when an infected male mates with an uninfected female, but the lethality is rescued if the female and her embryos are likewise infected. Two phage WO genes, cifAwMel and cifBwMel from the wMel Wolbachia deployed in vector control, transgenically recapitulate variably penetrant CI, and one of the same genes, cifAwMel, rescues wild type CI. The proposed Two-by-One genetic model predicts that CI and rescue can be recapitulated by transgenic expression alone and that dual cifAwMel and cifBwMel expression can recapitulate strong CI. Here, we use hatch rate and gene expression analyses in transgenic Drosophila melanogaster to demonstrate that CI and rescue can be synthetically recapitulated in full, and strong, transgenic CI comparable to wild type CI is achievable. These data explicitly validate the Two-by-One model in wMel-infected D. melanogaster, establish a robust system for transgenic studies of CI in a model system, and represent the first case of completely engineering male and female animal reproduction to depend upon bacteriophage gene products.}, } @article {pmid31238839, year = {2019}, author = {Ritchie, SA and Staunton, KM}, title = {Reflections from an old Queenslander: can rear and release strategies be the next great era of vector control?.}, journal = {Proceedings. Biological sciences}, volume = {286}, number = {1905}, pages = {20190973}, pmid = {31238839}, issn = {1471-2954}, mesh = {Aedes ; Animals ; Dengue ; *Mosquito Control ; *Mosquito Vectors ; *Pest Control, Biological ; Queensland ; Wolbachia ; Zika Virus Infection ; }, abstract = {In this perspective, I discuss the great eras of vector control, centring on Aedes aegypti, the primary vector of dengue, Zika and several other viruses. Since the discovery and acceptance of the role of mosquitoes as vectors of disease agents, several significant strategies have been developed and deployed to control them and the diseases they transmit. Environmental management, insecticides and, to a lesser extent, biological control have emerged as great eras of vector control. In the past decade, the release of massive numbers of specifically modified mosquitoes that mate with wild populations has emerged as a significant new strategy to fight vector-borne diseases. These reared and released mosquitoes have been modified by the addition of a symbiont (e.g. Wolbachia bacteria), radiation or introduction of a genetic construct to either sterilize the wild mosquitoes they mate with, crashing the population, or to reduce the wild population's capacity to vector pathogens. Will these new rear and release strategies become the next great era of vector control? From my vantage point as a dengue control manager and researcher involved in two Wolbachia programmes, I will discuss the hurdles that rear and release programmes face to gain widespread acceptance and success.}, } @article {pmid31234915, year = {2019}, author = {Zueva, T and Morchón, R and Carretón, E and Montoya-Alonso, JA and Santana, A and Bargues, MD and Mas-Coma, S and Rodríguez-Barbero, A and Simón, F}, title = {Angiogenic response in an in vitro model of dog microvascular endothelial cells stimulated with antigenic extracts from Dirofilaria immitis adult worms.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {315}, pmid = {31234915}, issn = {1756-3305}, support = {Castilla y León//Agencia de Desarrollo Económico/ ; Spain (cofinanced with FEDER funds)//Agencia de Desarrollo Económico/ ; Project No. RD16/0027/0018//RICET Red de Investigación de Centros de Enfermedades Tropicales/ ; Project No. RD16/0027/0023//Financial support for Valencia team obtained by RICET/ ; }, mesh = {Animals ; Antigens, Bacterial/pharmacology ; Antigens, Helminth/*pharmacology ; Capillaries/drug effects ; Cell Survival/drug effects ; Cells, Cultured ; Dirofilaria immitis/*chemistry/microbiology ; Dogs ; Endothelial Cells/*drug effects ; Inflammation ; Neovascularization, Physiologic/*drug effects ; Wolbachia/chemistry/genetics ; }, abstract = {BACKGROUND: Angiogenesis can occur under pathological conditions when stimuli such as inflammation, vascular obstruction or hypoxia exist. These stimuli are present in cardiopulmonary dirofilariosis (Dirofilaria immitis). The aim of this study was to analyze the capacity of D. immitis antigens to modify the expression of angiogenic factors and trigger the formation of pseudocapillaries (tube-like structures) in an in vitro model of endothelial cells.

METHODS: The expression of VEGF-A, sFlt, mEndoglin and sEndoglin in cultures of canine microvascular endothelial cells stimulated with extract of adult worms of D. immitis obtained from an untreated dog (DiSA) and from a dog treated for 15 days with doxycycline (tDiSA), was determined by using commercial kits. The capacity of pseudocapillary formation was evaluated analyzing cell connections and cell groups in Matrigel cell cultures stimulated with DiSA and tDiSA. In both cases non-stimulated cultures were used as controls.

RESULTS: First, we demonstrated that worms obtained from the dog treated with doxycycline showed a significantly lower amount of Wolbachia (less than 60%) than worms removed from the untreated dog. Only DiSA was able to significantly increase the expression of the proangiogenic factor VEGF-A in the endotelial cells cultures. None of the D. immitis extracts modified the expression of sFlt. tDiSA extract was able to modify the expression of the endoglins, significantly decreasing the expression of the pro-angiogenic mEndoglin and increasing the anti-angiogenic sEndoglin. The formation of pseudocapillaries was negatively influenced by tDiSA, which reduced the organization and number of cellular connections.

CONCLUSIONS: The ability of antigens from adult D. immitis worms to modify the expression of pro and anti-angiogenic factors in endotelial cell cultures was demonstrated, as well as the trend to form pseudocapillaries in vitro. The capacity of stimulation may be linked to the amount of Wolbachia present in the antigenic extracts.}, } @article {pmid31234774, year = {2019}, author = {Thapa, S and Zhang, Y and Allen, MS}, title = {Bacterial microbiomes of Ixodes scapularis ticks collected from Massachusetts and Texas, USA.}, journal = {BMC microbiology}, volume = {19}, number = {1}, pages = {138}, pmid = {31234774}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; Dogs/microbiology ; Female ; High-Throughput Nucleotide Sequencing/*methods ; Ixodes/*microbiology ; Male ; Massachusetts ; Microbiota ; Phylogeny ; Phylogeography ; RNA, Ribosomal, 16S/*genetics ; Sequence Analysis, DNA/methods ; Sex Characteristics ; Texas ; }, abstract = {BACKGROUND: The blacklegged tick, Ixodes scapularis, is the primary vector of the Lyme disease spirochete Borrelia burgdorferi in North America. Though the tick is found across the eastern United States, Lyme disease is endemic to the northeast and upper midwest and rare or absent in the southern portion of the vector's range. In an effort to better understand the tick microbiome from diverse geographic and climatic regions, we analysed the bacterial community of 115 I. scapularis adults collected from vegetation in Texas and Massachusetts, representing extreme ends of the vector's range, by massively parallel sequencing of the 16S V4 rRNA gene. In addition, 7 female I. scapularis collected from dogs in Texas were included in the study.

RESULTS: Male I. scapularis ticks had a more diverse bacterial microbiome in comparison to the female ticks. Rickettsia spp. dominated the microbiomes of field-collected female I. scapularis from both regions, as well as half of the males from Texas. In addition, the male and female ticks captured from Massachusetts contained high proportions of the pathogens Anaplasma and Borrelia, as well as the arthropod endosymbiont Wolbachia. None of these were found in libraries generated from ticks collected in Texas. Pseudomonas, Acinetobacter and Mycobacterium were significantly differently abundant (p < 0.05) between the male ticks from Massachusetts and Texas. Anaplasma and Borrelia were found in 15 and 63% of the 62 Massachusetts ticks, respectively, with a co-infection rate of 11%. Female ticks collected from Texas dogs were particularly diverse, and contained several genera including Rickettsia, Pseudomonas, Bradyrhizobium, Sediminibacterium, and Ralstonia.

CONCLUSIONS: Our results indicate that the bacterial microbiomes of I. scapularis ticks vary by sex and geography, with significantly more diversity in male microbiomes compared to females. We found that sex plays a larger role than geography in shaping the composition/diversity of the I. scapularis microbiome, but that geography affects what additional taxa are represented (beyond Rickettsia) and whether pathogens are found. Furthermore, recent feeding may have a role in shaping the tick microbiome, as evident from a more complex bacterial community in female ticks from dogs compared to the wild-caught questing females. These findings may provide further insight into the differences in the ability of the ticks to acquire, maintain and transmit pathogens. Future studies on possible causes and consequences of these differences will shed additional light on tick microbiome biology and vector competence.}, } @article {pmid31233646, year = {2019}, author = {Hinojosa, JC and Koubínová, D and Szenteczki, MA and Pitteloud, C and Dincă, V and Alvarez, N and Vila, R}, title = {A mirage of cryptic species: Genomics uncover striking mitonuclear discordance in the butterfly Thymelicus sylvestris.}, journal = {Molecular ecology}, volume = {28}, number = {17}, pages = {3857-3868}, doi = {10.1111/mec.15153}, pmid = {31233646}, issn = {1365-294X}, mesh = {Animals ; Bayes Theorem ; Butterflies/*genetics ; Cell Nucleus/*genetics ; Electron Transport Complex IV/genetics ; Genetic Loci ; *Genomics ; Likelihood Functions ; Mitochondria/*genetics ; Phylogeny ; Polymorphism, Single Nucleotide/genetics ; Species Specificity ; }, abstract = {Mitochondrial DNA (mtDNA) sequencing has led to an unprecedented rise in the identification of cryptic species. However, it is widely acknowledged that nuclear DNA (nuDNA) sequence data are also necessary to properly define species boundaries. Next generation sequencing techniques provide a wealth of nuclear genomic data, which can be used to ascertain both the evolutionary history and taxonomic status of putative cryptic species. Here, we focus on the intriguing case of the butterfly Thymelicus sylvestris (Lepidoptera: Hesperiidae). We identified six deeply diverged mitochondrial lineages; three distributed all across Europe and found in sympatry, suggesting a potential case of cryptic species. We then sequenced these six lineages using double-digest restriction-site associated DNA sequencing (ddRADseq). Nuclear genomic loci contradicted mtDNA patterns and genotypes generally clustered according to geography, i.e., a pattern expected under the assumption of postglacial recolonization from different refugia. Further analyses indicated that this strong mtDNA/nuDNA discrepancy cannot be explained by incomplete lineage sorting, sex-biased asymmetries, NUMTs, natural selection, introgression or Wolbachia-mediated genetic sweeps. We suggest that this mitonuclear discordance was caused by long periods of geographic isolation followed by range expansions, homogenizing the nuclear but not the mitochondrial genome. These results highlight T. sylvestris as a potential case of multiple despeciation and/or lineage fusion events. We finally argue, since mtDNA and nuDNA do not necessarily follow the same mechanisms of evolution, their respective evolutionary history reflects complementary aspects of past demographic and biogeographic events.}, } @article {pmid31229556, year = {2020}, author = {Parvathi, A and Jasna, V and Aswathy, VK and Aparna, S and Nathan, VK and Jyothibabu, R}, title = {Dominance of Wolbachia sp. in the deep-sea sediment bacterial metataxonomic sequencing analysis in the Bay of Bengal, Indian Ocean.}, journal = {Genomics}, volume = {112}, number = {1}, pages = {1030-1041}, doi = {10.1016/j.ygeno.2019.06.019}, pmid = {31229556}, issn = {1089-8646}, mesh = {Alphaproteobacteria/isolation & purification ; Bacteria/classification/genetics/isolation & purification ; Biodiversity ; Firmicutes/isolation & purification ; Geologic Sediments/chemistry/*microbiology ; High-Throughput Nucleotide Sequencing ; Indian Ocean ; Metagenomics ; Oceans and Seas ; Proteobacteria/isolation & purification ; Sequence Analysis, DNA ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The Bay of Bengal, located in the north-eastern part of the Indian Ocean is world's largest bay occupying an area of ~8,39,000 mile[2]. The variability in bacterial community structure and function in sediment ecosystems of the Bay of Bengal is examined by Illumina high-throughput metagenomic sequencing. Of five metataxonomics data sets presented, two (SD1 and SD2) were from stations close to the shore and three (SD4, SD5, and SD6) were from the deep-sea (~3000 m depth). Phylum Proteobacteria (90.27 to 92.52%) dominated the deep-sea samples, whereas phylum Firmicutes (65.35 to 90.98%) dominated the coastal samples. Comparative analysis showed that coastal and deep-sea sediments showed distinct microbial communities. Wolbachia species, belonging to class Alphaproteobacteria was the most dominant species in the deep-sea sediments. The gene functions of bacterial communities were predicted for deep-sea and coastal sediment ecosystems. The results indicated that deep-sea sediment bacterial communities were involved in metabolic activities like dehalogenation and sulphide oxidation.}, } @article {pmid31229468, year = {2019}, author = {Bock, W and Jayathunga, Y}, title = {Optimal control of a multi-patch Dengue model under the influence of Wolbachia bacterium.}, journal = {Mathematical biosciences}, volume = {315}, number = {}, pages = {108219}, doi = {10.1016/j.mbs.2019.108219}, pmid = {31229468}, issn = {1879-3134}, mesh = {Aedes/*microbiology ; Animals ; *Basic Reproduction Number ; Dengue/*prevention & control/*transmission ; Humans ; *Models, Biological ; *Mosquito Vectors ; *Pest Control, Biological ; *Wolbachia/pathogenicity ; }, abstract = {In this work, a multi-patch model for dengue transmission dynamics including the bacterium Wolbachia is studied and by that the control efforts to minimize the disease spread by host and vector control are investigated. The multi-patch system models the host movement within the patches which coupled via a residence-time budgeting matrix P. Numerical results confirm that the control mechanism embedded in incidence rates of the disease transmission, effectively reduce the spread of the disease.}, } @article {pmid31227544, year = {2019}, author = {Cooper, BS and Vanderpool, D and Conner, WR and Matute, DR and Turelli, M}, title = {Wolbachia Acquisition by Drosophila yakuba-Clade Hosts and Transfer of Incompatibility Loci Between Distantly Related Wolbachia.}, journal = {Genetics}, volume = {212}, number = {4}, pages = {1399-1419}, pmid = {31227544}, issn = {1943-2631}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; R01 GM121750/GM/NIGMS NIH HHS/United States ; R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*genetics/microbiology/physiology ; Female ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Genome, Insect ; Genome, Mitochondrial ; Host-Pathogen Interactions ; Infertility/*genetics ; Male ; Phylogeny ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Maternally transmitted Wolbachia infect about half of insect species, yet the predominant mode(s) of Wolbachia acquisition remains uncertain. Species-specific associations could be old, with Wolbachia and hosts codiversifying (i.e., cladogenic acquisition), or relatively young and acquired by horizontal transfer or introgression. The three Drosophila yakuba-clade hosts [(D. santomea, D. yakuba) D. teissieri] diverged ∼3 MYA and currently hybridize on the West African islands Bioko and São Tomé. Each species is polymorphic for nearly identical Wolbachia that cause weak cytoplasmic incompatibility (CI)-reduced egg hatch when uninfected females mate with infected males. D. yakuba-clade Wolbachia are closely related to wMel, globally polymorphic in D. melanogaster We use draft Wolbachia and mitochondrial genomes to demonstrate that D. yakuba-clade phylogenies for Wolbachia and mitochondria tend to follow host nuclear phylogenies. However, roughly half of D. santomea individuals, sampled both inside and outside of the São Tomé hybrid zone, have introgressed D. yakuba mitochondria. Both mitochondria and Wolbachia possess far more recent common ancestors than the bulk of the host nuclear genomes, precluding cladogenic Wolbachia acquisition. General concordance of Wolbachia and mitochondrial phylogenies suggests that horizontal transmission is rare, but varying relative rates of molecular divergence complicate chronogram-based statistical tests. Loci that cause CI in wMel are disrupted in D. yakuba-clade Wolbachia; but a second set of loci predicted to cause CI are located in the same WO prophage region. These alternative CI loci seem to have been acquired horizontally from distantly related Wolbachia, with transfer mediated by flanking Wolbachia-specific ISWpi1 transposons.}, } @article {pmid31222094, year = {2019}, author = {Mateos, M and Silva, NO and Ramirez, P and Higareda-Alvear, VM and Aramayo, R and Erickson, JW}, title = {Effect of heritable symbionts on maternally-derived embryo transcripts.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {8847}, pmid = {31222094}, issn = {2045-2322}, support = {R03 AI078348/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*embryology/genetics/*microbiology ; Embryo, Nonmammalian/*microbiology ; Female ; Genes, Insect/genetics ; Host-Pathogen Interactions/genetics ; Male ; Phenotype ; RNA, Ribosomal ; Reproduction/genetics ; Ribosome Inactivating Proteins/genetics/physiology ; Sequence Analysis, RNA ; Spiroplasma/enzymology ; *Symbiosis ; Transcriptome/*genetics ; Wolbachia ; }, abstract = {Maternally-transmitted endosymbiotic bacteria are ubiquitous in insects. Among other influential phenotypes, many heritable symbionts of arthropods are notorious for manipulating host reproduction through one of four reproductive syndromes, which are generally exerted during early developmental stages of the host: male feminization; parthenogenesis induction; male killing; and cytoplasmic incompatibility (CI). Major advances have been achieved in understanding mechanisms and identifying symbiont factors involved in reproductive manipulation, particularly male killing and cytoplasmic incompatibility. Nonetheless, whether cytoplasmically-transmitted bacteria influence the maternally-loaded components of the egg or early embryo has not been examined. In the present study, we investigated whether heritable endosymbionts that cause different reproductive phenotypes in Drosophila melanogaster influence the mRNA transcriptome of early embryos. We used mRNA-seq to evaluate differential expression in Drosophila embryos lacking endosymbionts (control) to those harbouring the male-killing Spiroplasma poulsonii strain MSRO-Br, the CI-inducing Wolbachia strain wMel, or Spiroplasma poulsonii strain Hyd1; a strain that lacks a reproductive phenotype and is naturally associated with Drosophila hydei. We found no consistent evidence of influence of symbiont on mRNA composition of early embryos, suggesting that the reproductive manipulation mechanism does not involve alteration of maternally-loaded transcripts. In addition, we capitalized on several available mRNA-seq datasets derived from Spiroplasma-infected Drosophila melanogaster embryos, to search for signals of depurination of rRNA, consistent with the activity of Ribosome Inactivating Proteins (RIPs) encoded by Spiroplasma poulsonii. We found small but statistically significant signals of depurination of Drosophila rRNA in the Spiroplasma treatments (both strains), but not in the symbiont-free control or Wolbachia treatment, consistent with the action of RIPs. The depurination signal was slightly stronger in the treatment with the male-killing strain. This result supports a recent report that RIP-induced damage contributes to male embryo death.}, } @article {pmid31215133, year = {2020}, author = {Su, QC and Wang, X and Deng, C and Yun, YL and Zhao, Y and Peng, Y}, title = {Transcriptome responses to elevated CO2 level and Wolbachia-infection stress in Hylyphantes graminicola (Araneae: Linyphiidae).}, journal = {Insect science}, volume = {27}, number = {5}, pages = {908-920}, doi = {10.1111/1744-7917.12701}, pmid = {31215133}, issn = {1744-7917}, support = {2016YFD0200900//State's Key Project of Research and Development Plan/ ; 31672317//National Natural Science Foundation of China/ ; 2016jzxjh012//Competitive Planning Projects of Hubei Academy of Agricultural Sciences/ ; }, mesh = {Animals ; Carbon Dioxide/*physiology ; Female ; Spiders/drug effects/*genetics/microbiology ; Stress, Physiological ; Transcriptome/drug effects/*genetics ; Wolbachia/*physiology ; }, abstract = {Hylyphantes graminicola is a resident spider species found in maize and cotton fields and is an important biological control agent of various pests. Previous studies have demonstrated that stress from elevated CO2 and Wolbachia infection can strongly affect spider species. Thus, based on CO2 levels (400 ppm, current atmospheric CO2 concentration and 800 ppm, high CO2 concentration) and Wolbachia status (Wolbachia-infected, W[+] and Wolbachia-uninfected, W[-]), we divided H. graminicola individuals into four treatment groups: W[-] 400 ppm, W[-] 800 ppm, W[+] 400 ppm, and W[+] 800 ppm. To investigate the effects of elevated CO2 levels (W[-] 400 vs W[-] 800), Wolbachia infection (W[-] 400 vs W[+] 400), and the interactions between these two factors (W[-] 400 vs W[+] 800), high-throughput transcriptome sequencing was employed to characterize the de novo transcriptome of the spiders and identify stress-related differentially expressed genes (DEGs). De novo assembly of complementary DNA sequences generated 86 688 unigenes, 23 938 of which were annotated in public databases. A total of 84, 21, and 157 DEGs were found among W[-] 400 vs W[-] 800, W[-] 400 vs W[+] 400, and W[-] 400 vs W[+] 800, respectively. Functional enrichment analysis revealed that metabolic processes, signaling, and catalytic activity were significantly affected by elevated CO2 levels and Wolbachia infection. Our findings suggest that the impact of elevated CO2 levels and Wolbachia infection on the H. graminicola transcriptome was, to a large extent, on genes involved in metabolic processes. This study is the first description of transcriptome changes in response to elevated CO2 levels and Wolbachia infection in spiders.}, } @article {pmid31214820, year = {2019}, author = {Konecka, E and Olszanowski, Z}, title = {First Evidence of Intracellular Bacteria Cardinium in Thermophilic Mite Microzetorchestes emeryi (Acari: Oribatida): Molecular Screening of Bacterial Endosymbiont Species.}, journal = {Current microbiology}, volume = {76}, number = {9}, pages = {1038-1044}, pmid = {31214820}, issn = {1432-0991}, mesh = {Animals ; Bacteroidetes/classification/*genetics/*isolation & purification/physiology ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Mites/*microbiology/physiology ; Phylogeny ; Poland ; Symbiosis ; }, abstract = {We undertook the issue of the distribution of intracellular bacteria among Oribatida (Acari). Six genera of bacteria were detected by PCR and Sanger DNA sequencing: Wolbachia, Cardinium, Rickettsia, Spiroplasma, Arsenophonus, and Hamiltonella. Our research, for the first time, revealed the presence of Cardinium in Microzetorchestes emeryi in two subpopulations separated from each other by 300 m. The percentages of infected animals were the same in both subpopulations-ca. 20%. The identity of 16S rDNA sequences of Cardinium between these two subpopulations of M. emeryi was 97%. Phylogenetic analysis showed that the Cardinium in M. emeryi was clustered into the group A. The occurrence of M. emeryi in Poland has not been reported before and our report is the first one. Cardinium maybe help the thermophilic M. emeryi to adapt to low temperatures in the Central Europe.}, } @article {pmid31208002, year = {2019}, author = {Schebeck, M and Feldkirchner, L and Stauffer, C and Schuler, H}, title = {Dynamics of an Ongoing Wolbachia Spread in the European Cherry Fruit Fly, Rhagoletis cerasi (Diptera: Tephritidae).}, journal = {Insects}, volume = {10}, number = {6}, pages = {}, pmid = {31208002}, issn = {2075-4450}, support = {J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 31441/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Numerous terrestrial arthropods are infected with the alphaproteobacterium Wolbachia. This endosymbiont is usually transmitted vertically from infected females to their offspring and can alter the reproduction of hosts through various manipulations, like cytoplasmic incompatibility (CI), enhancing its spread in new host populations. Studies on the spatial and temporal dynamics of Wolbachia under natural conditions are scarce. Here, we analyzed Wolbachia infection frequencies in populations of the European cherry fruit fly, Rhagoletis cerasi (L.), in central Germany-an area of an ongoing spread of the CI-inducing strain wCer2. In total, 295 individuals from 19 populations were PCR-screened for the presence of wCer2 and their mitochondrial haplotype. Results were compared with historic data to understand the infection dynamics of the ongoing wCer2 invasion. An overall wCer2 infection frequency of about 30% was found, ranging from 0% to 100% per population. In contrast to an expected smooth transition from wCer2-infected to completely wCer2-uninfected populations, a relatively scattered infection pattern across geography was observed. Moreover, a strong Wolbachia-haplotype association was detected, with only a few rare misassociations. Our results show a complex dynamic of an ongoing Wolbachia spread in natural field populations of R. cerasi.}, } @article {pmid31204356, year = {2019}, author = {Watanabe, A and Takaku, S and Yokota, K and Hayashi, S and Tamaki, N and Kokeguchi, S}, title = {A survey of Lasioderma serricorne (Fabricius) in Japanese Dental Clinics.}, journal = {Biocontrol science}, volume = {24}, number = {2}, pages = {117-121}, doi = {10.4265/bio.24.117}, pmid = {31204356}, issn = {1884-0205}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; Cluster Analysis ; Coleoptera/*microbiology ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; *Dental Clinics ; Japan ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {This study was to survey the capturing rate in Japanese dental clinics of the Lasioderma serricorne (cigarette beetles) , and to evaluate the beetle's potential as a carrier for transmission of nosocomial pathogens. L. serricorne imagoes were captured in pheromone traps in 14 Japanese dental clinics in August and September 2012 and 2013, and their numbers recorded. Polymerase chain reaction (PCR) for the bacterial antibiotic-resistant genes mecA, vanA, vanB, blaIMP, and blaVIM was performed on the captured L. serricorne imagoes. Bacterial species in the captured specimens were identified by 16S rRNA PCR and sequencing analysis. The L. serricorne imagoes were captured from 10 dental clinics (71.4%) . We failed to detect the presence of nosocomial antibiotic-resistant pathogens in L. serricorne imagoes. The bacterial species detected most commonly in the imagoes was Wolbachia sp., an intracellular proteobacterium infecting certain insect species. Monitoring of insects including L. serricorne should be incorporated into regiment of the infection control.}, } @article {pmid31194893, year = {2020}, author = {Ant, TH and Herd, C and Louis, F and Failloux, AB and Sinkins, SP}, title = {Wolbachia transinfections in Culex quinquefasciatus generate cytoplasmic incompatibility.}, journal = {Insect molecular biology}, volume = {29}, number = {1}, pages = {1-8}, pmid = {31194893}, issn = {1365-2583}, support = {202888/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; BB/K004506/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Aedes/microbiology ; Animals ; Culex/*microbiology ; Female ; Male ; Mosquito Control/methods ; Mosquito Vectors/microbiology ; Pest Control, Biological/methods ; Salivary Glands/microbiology ; Symbiosis ; Wolbachia/classification/*physiology ; }, abstract = {Culex quinquefasciatus is an important mosquito vector of a number of viral and protozoan pathogens of humans and animals, and naturally carries the endosymbiont Wolbachia pipientis, strain wPip. Wolbachia are used in two distinct vector control strategies: firstly, population suppression caused by mating incompatibilities between mass-released transinfected males and wild females; and secondly, the spread of pathogen transmission-blocking strains through populations. Using embryonic microinjection, two novel Wolbachia transinfections were generated in C. quinquefasciatus using strains native to the mosquito Aedes albopictus: a wAlbB single infection, and a wPip plus wAlbA superinfection. The wAlbB infection showed full bidirectional cytoplasmic incompatibility (CI) with wild-type C. quinquefasciatus in reciprocal crosses. The wPipwAlbA superinfection showed complete unidirectional CI, and therefore population invasion potential. Whereas the wAlbB strain showed comparatively low overall densities, similar to the native wPip, the wPipwAlbA superinfection reached over 400-fold higher densities in the salivary glands compared to the native wPip, suggesting it may be a candidate for pathogen transmission blocking.}, } @article {pmid31191980, year = {2019}, author = {Koh, C and Audsley, MD and Di Giallonardo, F and Kerton, EJ and Young, PR and Holmes, EC and McGraw, EA}, title = {Sustained Wolbachia-mediated blocking of dengue virus isolates following serial passage in Aedes aegypti cell culture.}, journal = {Virus evolution}, volume = {5}, number = {1}, pages = {vez012}, pmid = {31191980}, issn = {2057-1577}, abstract = {Wolbachia is an intracellular endosymbiont of insects that inhibits the replication of a range of pathogens in its arthropod hosts. The release of Wolbachia into wild populations of mosquitoes is an innovative biocontrol effort to suppress the transmission of arthropod-borne viruses (arboviruses) to humans, most notably dengue virus. The success of the Wolbachia-based approach hinges upon the stable persistence of the 'pathogen blocking' effect, whose mechanistic basis is poorly understood. Evidence suggests that Wolbachia may affect viral replication via a combination of competition for host resources and activation of host immunity. The evolution of resistance against Wolbachia and pathogen blocking in the mosquito or the virus could reduce the public health impact of the symbiont releases. Here, we investigate if dengue 3 virus (DENV-3) is capable of accumulating adaptive mutations that improve its replicative capacity during serial passage in Wolbachia wMel-infected cells. During the passaging regime, viral isolates in Wolbachia-infected cells exhibited greater variation in viral loads compared to controls. The viral loads of these isolates declined rapidly during passaging due to the blocking effects of Wolbachia carriage, with several being lost all together and the remainder recovering to low but stable levels. We attempted to sequence the genomes of the surviving passaged isolates but, given their low abundance, were unable to obtain sufficient depth of coverage for evolutionary analysis. In contrast, viral loads in Wolbachia-free control cells were consistently high during passaging. The surviving isolates passaged in the presence of Wolbachia exhibited a reduced ability to replicate even in Wolbachia-free cells. These experiments demonstrate the challenge for dengue in evolving resistance to Wolbachia-mediated blocking.}, } @article {pmid31186318, year = {2019}, author = {Chrostek, E and Gerth, M}, title = {Is Anopheles gambiae a Natural Host of Wolbachia?.}, journal = {mBio}, volume = {10}, number = {3}, pages = {}, pmid = {31186318}, issn = {2150-7511}, mesh = {Africa ; Animals ; Anopheles/*microbiology ; Female ; Genome, Bacterial ; Malaria/transmission ; Male ; Metagenomics ; Mosquito Vectors/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Wolbachia (Alphaproteobacteria, Rickettsiales) is an intraovarially transmitted symbiont of insects able to exert striking phenotypes, including reproductive manipulations and pathogen blocking. These phenotypes make Wolbachia a promising tool to combat mosquito-borne diseases. Although Wolbachia is present in the majority of terrestrial arthropods, including many disease vectors, it was considered absent from Anopheles gambiae mosquitos, the main vectors of malaria in sub-Saharan Africa. In 2014, Wolbachia sequences were detected in A. gambiae samples collected in Burkina Faso. Subsequently, similar evidence came from collections all over Africa, revealing a high Wolbachia 16S rRNA sequence diversity, low abundance, and a lack of congruence between host and symbiont phylogenies. Here, we reanalyze and discuss recent evidence on the presence of Wolbachia sequences in A. gambiae. We find that although detected at increasing frequencies, the unusual properties of these Wolbachia sequences render them insufficient to diagnose natural infections in A. gambiae Future studies should focus on uncovering the origin of Wolbachia sequence variants in Anopheles and seeking sequence-independent evidence for this new symbiosis. Understanding the ecology of Anopheles mosquitos and their interactions with Wolbachia will be key in designing successful, integrative approaches to limit malaria spread. Although the prospect of using Wolbachia to fight malaria is intriguing, the newly discovered strains do not bring it closer to realization.IMPORTANCEAnopheles gambiae mosquitos are the main vectors of malaria, threatening around half of the world's population. The bacterial symbiont Wolbachia can interfere with disease transmission by other important insect vectors, but until recently, it was thought to be absent from natural A. gambiae populations. Here, we critically analyze the genomic, metagenomic, PCR, imaging, and phenotypic data presented in support of the presence of natural Wolbachia infections in A. gambiae We find that they are insufficient to diagnose Wolbachia infections and argue for the need of obtaining robust data confirming basic Wolbachia characteristics in this system. Determining the Wolbachia infection status of Anopheles is critical due to its potential to influence Anopheles population structure and Plasmodium transmission.}, } @article {pmid31176069, year = {2019}, author = {Caragata, EP and Tikhe, CV and Dimopoulos, G}, title = {Curious entanglements: interactions between mosquitoes, their microbiota, and arboviruses.}, journal = {Current opinion in virology}, volume = {37}, number = {}, pages = {26-36}, pmid = {31176069}, issn = {1879-6265}, support = {R01 AI078997/AI/NIAID NIH HHS/United States ; R21 AI136456/AI/NIAID NIH HHS/United States ; R01 AI141532/AI/NIAID NIH HHS/United States ; R01 AI081877/AI/NIAID NIH HHS/United States ; R01 AI121253/AI/NIAID NIH HHS/United States ; R21 AI131574/AI/NIAID NIH HHS/United States ; R01 AI061576/AI/NIAID NIH HHS/United States ; R01 AI122743/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/immunology/microbiology/virology ; Animals ; Anopheles/immunology/microbiology/virology ; Arbovirus Infections/immunology/microbiology/virology ; Arboviruses/pathogenicity ; Biological Control Agents/immunology ; Culex/immunology/microbiology/virology ; Dengue Virus/pathogenicity ; Host Microbial Interactions/*immunology ; Humans ; Immunity, Innate ; Microbial Interactions/*immunology ; Microbiota ; *Mosquito Vectors/immunology/microbiology/virology ; Virus Diseases/immunology/microbiology/virology ; Wolbachia/*immunology ; }, abstract = {Mosquitoes naturally harbor a diverse community of microorganisms that play a crucial role in their biology. Mosquito-microbiota interactions are abundant and complex. They can dramatically alter the mosquito immune response, and impede or enhance a mosquito's ability to transmit medically important arboviral pathogens. Yet critically, given the massive public health impact of arboviral disease, few such interactions have been well characterized. In this review, we describe the current state of knowledge of the role of microorganisms in mosquito biology, how microbial-induced changes to mosquito immunity moderate infection with arboviruses, cases of mosquito-microbial-virus interactions with a defined mechanism, and the molecular interactions that underlie the endosymbiotic bacterium Wolbachia's ability to block virus infection in mosquitoes.}, } @article {pmid31174466, year = {2019}, author = {Baião, GC and Schneider, DI and Miller, WJ and Klasson, L}, title = {The effect of Wolbachia on gene expression in Drosophila paulistorum and its implications for symbiont-induced host speciation.}, journal = {BMC genomics}, volume = {20}, number = {1}, pages = {465}, pmid = {31174466}, issn = {1471-2164}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; 2014-4353//Vetenskapsrådet/ ; P28255-B22//Austrian Science Fund/ ; }, mesh = {Amino Acids/metabolism ; Animals ; Carbohydrate Metabolism/genetics ; Drosophila/*genetics/metabolism/*microbiology ; Female ; *Genetic Speciation ; Immunity/genetics ; Lipid Metabolism/genetics ; Male ; Muscles/metabolism ; Pheromones/metabolism ; Proteolysis ; RNA-Seq ; Reproduction/genetics ; *Symbiosis ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The Neotropical fruit fly Drosophila paulistorum (Diptera: Drosophilidae) is a species complex in statu nascendi comprising six reproductively isolated semispecies, each harboring mutualistic Wolbachia strains. Although wild type flies of each semispecies are isolated from the others by both pre- and postmating incompatibilities, mating between semispecies and successful offspring development can be achieved once flies are treated with antibiotics to reduce Wolbachia titer. Here we use RNA-seq to study the impact of Wolbachia on D. paulistorum and investigate the hypothesis that the symbiont may play a role in host speciation. For that goal, we analyze samples of heads and abdomens of both sexes of the Amazonian, Centro American and Orinocan semispecies of D. paulistorum.

RESULTS: We identify between 175 and 1192 differentially expressed genes associated with a variety of biological processes that respond either globally or according to tissue, sex or condition in the three semispecies. Some of the functions associated with differentially expressed genes are known to be affected by Wolbachia in other species, such as metabolism and immunity, whereas others represent putative novel phenotypes involving muscular functions, pheromone signaling, and visual perception.

CONCLUSIONS: Our results show that Wolbachia affect a large number of biological functions in D. paulistorum, particularly when present in high titer. We suggest that the significant metabolic impact of the infection on the host may cause several of the other putative and observed phenotypes. We also speculate that the observed differential expression of genes associated with chemical communication and reproduction may be associated with the emergence of pre- and postmating barriers between semispecies, which supports a role for Wolbachia in the speciation of D. paulistorum.}, } @article {pmid31173475, year = {2020}, author = {Xie, K and Lu, YJ and Yang, K and Huo, SM and Hong, XY}, title = {Co-infection of Wolbachia and Spiroplasma in spider mite Tetranychus truncatus increases male fitness.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {921-937}, pmid = {31173475}, issn = {1744-7917}, support = {31672035//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; }, mesh = {Animals ; *Genetic Fitness ; Male ; Spiroplasma/*physiology ; *Symbiosis ; Tetranychidae/*genetics/microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia and Spiroplasma are intracellular bacteria that are of great interest to entomologists, because of their ability to alter insect host biology in multiple ways. In the spider mite Tetranychus truncatus, co-infection of Wolbachia and Spiroplasma can induce cytoplasmic incompatibility (CI) and fitness costs; however, little is known about the effect of co-infection at the genetic level and the molecular mechanisms underlying CI. In this study, we explored the influence of the two symbionts on male mite host fitness and used RNA sequencing to generate the transcriptomes of T. truncatus with four different types of infection. In total, we found symbiont-infected lines had a higher hatch proportion than the uninfected line, and the development time of the uninfected line was longer than that of the other lines. Co-infection changed the expression of many genes related to digestion detoxification, reproduction, immunity and oxidation reduction. Our results indicate that co-infection of Wolbachia and Spiroplasma confers multiple effects on their hosts, and helps illuminate the complex interactions between endosymbionts and arthropods.}, } @article {pmid31167953, year = {2020}, author = {Bonning, BC}, title = {The Insect Virome: Opportunities and Challenges.}, journal = {Current issues in molecular biology}, volume = {34}, number = {}, pages = {1-12}, doi = {10.21775/cimb.034.001}, pmid = {31167953}, issn = {1467-3045}, mesh = {Animals ; Biodiversity ; Genome, Viral ; Host-Pathogen Interactions/genetics/immunology ; Insect Viruses/*classification/*genetics ; Insecta/genetics/immunology/*virology ; Invertebrates ; Metagenomics ; Microbiota ; }, abstract = {The insect virome is composed of a myriad of viruses. Both field populations and laboratory colonies of insects harbour diverse viruses, including viruses that infect the insect itself, viruses of microbes associated with the insect, and viruses associated with ingested materials. Metagenomics analysis for identification of virus-derived sequences has allowed for new appreciation of the extent and diversity of the insect virome. The complex interactions between insect viruses and host antiviral immune pathways (RNA interference and apoptosis), and between viruses and other members of the microbiome (e.g. Wolbachia) are becoming apparent. In this chapter, an overview of the diversity of viruses in insects and recent virus discovery research for specific insects and insect-derived cell lines is provided. The opportunities and challenges associated with the insect virome, including the potential impacts of viruses on both research and insect management programs are also addressed.}, } @article {pmid31162017, year = {2019}, author = {Batsa Debrah, L and Phillips, RO and Pfarr, K and Klarmann-Schulz, U and Opoku, VS and Nausch, N and Owusu, W and Mubarik, Y and Sander, AL and Lämmer, C and Ritter, M and Layland, LE and Jacobsen, M and Debrah, AY and Hoerauf, A}, title = {The Efficacy of Doxycycline Treatment on Mansonella perstans Infection: An Open-Label, Randomized Trial in Ghana.}, journal = {The American journal of tropical medicine and hygiene}, volume = {101}, number = {1}, pages = {84-92}, pmid = {31162017}, issn = {1476-1645}, support = {MR/J01477X/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/*therapeutic use ; Child ; Doxycycline/*therapeutic use ; Female ; Ghana/epidemiology ; Humans ; Male ; *Mansonella ; Mansonelliasis/*drug therapy/*epidemiology ; Middle Aged ; Young Adult ; }, abstract = {Treating Mansonella perstans is challenged by the low efficacy of registered antihelminthics. Wolbachia endobacteria provide an alternative treatment target because depletion results in amicrofilaremia in filarial infections with Wuchereria bancrofti and Onchocerca volvulus infections. This open-label, randomized study sought to confirm that i) Wolbachia are present in M. perstans in Ghana and ii) doxycycline treatment will deplete Wolbachia and cause a slow, sustained decline in microfilariae (MF). Two hundred and two Ghanaians with M. perstans infection were randomized into early (immediate) and delayed (6 months deferred) treatment groups, given doxycycline 200 mg/day for 6 weeks, and monitored for MF and Wolbachia levels at baseline, 4, 12, and 24 months after the study onset (= time of randomization and start of treatment for the early group). Per protocol analysis revealed that the median MF/mL in the early group declined from 138 at baseline to 64 at month 4 and further to 0 at month 12. In the delayed group, MF load did not change from a baseline median of 97 to 102 at month 4 but declined to 42 at month 12, that is, 6 months after receiving treatment, trailing the early group as expected. By month 24, both treatment groups had reached a median MF level of 0. After treatment, Wolbachia were depleted from MF by ≥ 1-log drop compared with baseline levels. We conclude that M. perstans in Ghana harbor Wolbachia that are effectively depleted by doxycycline with subsequent reduction in MF loads, most likely because of interruption of fertility of adult worms.}, } @article {pmid31161558, year = {2019}, author = {Li, D and Wan, H}, title = {The Threshold Infection Level for [Formula: see text] Invasion in a Two-Sex Mosquito Population Model.}, journal = {Bulletin of mathematical biology}, volume = {81}, number = {7}, pages = {2596-2624}, doi = {10.1007/s11538-019-00620-1}, pmid = {31161558}, issn = {1522-9602}, mesh = {Animals ; *Biological Control Agents ; Computer Simulation ; Culicidae/growth & development/*microbiology ; Female ; Humans ; Male ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/growth & development/*microbiology ; Symbiosis/physiology ; Vector Borne Diseases/prevention & control/transmission ; Wolbachia/*pathogenicity/*physiology ; }, abstract = {In this paper, we formulate a new [Formula: see text] infection model in a two-sex mosquito population with stage structure. Some key factors of [Formula: see text] infection, including cytoplasmic incompatibility (CI), male killing (MK) effect, maternal transmission, fecundity cost due to fitness effect and different mortality rates for infected individuals, are captured. Dynamical analysis has been carried out, and the basic reproduction number [Formula: see text] for [Formula: see text] infection has been calculated. Our analysis shows that [Formula: see text] can establish in a mosquito population if [Formula: see text] is greater than unity. If [Formula: see text] is less than unity, [Formula: see text] establishment still can be achieved if backward bifurcation occurs. Under this circumstance, the initial values lying in the basin of attraction of the stable [Formula: see text]-established equilibrium are essential to guarantee [Formula: see text] establishment. In particular, the method to find the basin of attraction and evaluate the threshold initial values is given. Besides, according to a comparison of different releasing strategies, it is shown that, from the perspective of economy and disease control, keeping the number of infected female mosquitoes to a necessary minimum by relying on higher number of male mosquitoes released is a desirable strategy. Moreover, global and local sensitivity analysis and numerical simulation have been performed to explore the impact of model parameters to the success of population establishment. Our results suggest that low levels of MK effect and fitness costs as well as high levels of CI and maternal inheritance are in favor of [Formula: see text] establishment. Moreover, not considering MK effect and incomplete CI effect may result in the underestimation of the number of infected mosquitoes needed to be released.}, } @article {pmid31161026, year = {2019}, author = {Kulkarni, A and Yu, W and Jiang, J and Sanchez, C and Karna, AK and Martinez, KJL and Hanley, KA and Buenemann, M and Hansen, IA and Xue, RD and Ettestad, P and Melman, S and Duguma, D and Debboun, M and Xu, J}, title = {Wolbachia pipientis occurs in Aedes aegypti populations in New Mexico and Florida, USA.}, journal = {Ecology and evolution}, volume = {9}, number = {10}, pages = {6148-6156}, pmid = {31161026}, issn = {2045-7758}, support = {P20 GM103451/GM/NIGMS NIH HHS/United States ; }, abstract = {The mosquitoes Aedes aegypti (L.) and Ae. albopictus Skuse are the major vectors of dengue, Zika, yellow fever, and chikungunya viruses worldwide. Wolbachia, an endosymbiotic bacterium present in many insects, is being utilized in novel vector control strategies to manipulate mosquito life history and vector competence to curb virus transmission. Earlier studies have found that Wolbachia is commonly detected in Ae. albopictus but rarely detected in Ae. aegypti. In this study, we used a two-step PCR assay to detect Wolbachia in wild-collected samples of Ae. aegypti. The PCR products were sequenced to validate amplicons and identify Wolbachia strains. A loop-mediated isothermal amplification (LAMP) assay was developed and used for detecting Wolbachia in selected mosquito specimens as well. We found Wolbachia in 85/148 (57.4%) wild Ae. aegypti specimens from various cities in New Mexico, and in 2/46 (4.3%) from St. Augustine, Florida. Wolbachia was not detected in 94 samples of Ae. aegypti from Deer Park, Harris County, Texas. Wolbachia detected in Ae. aegypti from both New Mexico and Florida was the wAlbB strain of Wolbachia pipientis. A Wolbachia-positive colony of Ae. aegypti was established from pupae collected in Las Cruces, New Mexico, in 2018. The infected females of this strain transmitted Wolbachia to their progeny when crossed with males of Rockefeller strain of Ae. aegypti, which does not carry Wolbachia. In contrast, none of the progeny of Las Cruces males mated to Rockefeller females were infected with Wolbachia.}, } @article {pmid31159272, year = {2019}, author = {Gichuhi, J and Khamis, FM and Van den Berg, J and Ekesi, S and Herren, JK}, title = {Unexpected Diversity of Wolbachia Associated with Bactrocera dorsalis (Diptera: Tephritidae) in Africa.}, journal = {Insects}, volume = {10}, number = {6}, pages = {}, pmid = {31159272}, issn = {2075-4450}, support = {IBCARP//European Union/ ; }, abstract = {Bactrocera dorsalis (Hendel) is an important pest of fruit-bearing plants in many countries worldwide. In Africa, this pest has spread rapidly and has become widely established since the first invasion report in 2003. Wolbachia is a vertically transmitted endosymbiont that can significantly influence aspects of the biology and, in particular, the reproduction of its host. In this study, we screened B. dorsalis specimens collected from several locations in Africa between 2005 and 2017 for Wolbachia using a PCR-based assay to target the Wolbachia surface protein wsp. Of the 357 individuals tested, 10 were positive for Wolbachia using the wsp assay. We identified four strains of Wolbachia infecting two B. dorsalis mitochondrial haplotypes. We found no strict association between the infecting strain and host haplotype, with one strain being present in two different host haplotypes. All the detected strains belonged to Super Group B Wolbachia and did not match any strains reported previously in B. dorsalis in Asia. These findings indicate that diverse Wolbachia infections are present in invasive populations of B. dorsalis.}, } @article {pmid31143363, year = {2019}, author = {Yashiro, T and Lo, N}, title = {Comparative screening of endosymbiotic bacteria associated with the asexual and sexual lineages of the termite Glyptotermes nakajimai.}, journal = {Communicative & integrative biology}, volume = {12}, number = {1}, pages = {55-58}, pmid = {31143363}, issn = {1942-0889}, abstract = {Males provide opportunities both for sexual reproduction and for sex-based phenotypic differences within animal societies. In termites, the ubiquitous presence of both male and female workers and soldiers indicate that males play a critical role in colonies of these insects. However, we have recently reported all-female asexual societies in a lineage of the termite Glyptotermes nakajimai - a dramatic transition from mixed-sex to all-female asexual societies. It is known that female-producing parthenogenesis in insects can be induced by maternally inherited endosymbiotic bacteria, such as Wolbachia, Cardinium, and Rickettsia. Here, we screen for the presence of endosymbiotic bacteria in the asexual and sexual lineages of G. nakajimai. Our bacterial screening of the asexual lineage did not reveal any likely causal agents for parthenogenetic reproduction, whereas screening of the sexual lineage resulted in Wolbachia being detected. Our findings suggest that the asexuality in G. nakajimai is likely to be maintained without manipulation by endosymbiotic bacteria.}, } @article {pmid31134014, year = {2019}, author = {Wasala, SK and Brown, AMV and Kang, J and Howe, DK and Peetz, AB and Zasada, IA and Denver, DR}, title = {Variable Abundance and Distribution of Wolbachia and Cardinium Endosymbionts in Plant-Parasitic Nematode Field Populations.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {964}, pmid = {31134014}, issn = {1664-302X}, abstract = {The bacterial endosymbiont Wolbachia interacts with different invertebrate hosts, engaging in diverse symbiotic relationships. Wolbachia is often a reproductive parasite in arthropods, but an obligate mutualist in filarial nematodes. Wolbachia was recently discovered in plant-parasitic nematodes, and, is thus far known in just two genera Pratylenchus and Radopholus, yet the symbiont's function remains unknown. The occurrence of Wolbachia in these economically important plant pests offers an unexplored biocontrol strategy. However, development of Wolbachia-based biocontrol requires an improved understanding of symbiont-host functional interactions and the symbiont's prevalence among nematode field populations. This study used a molecular-genetic approach to assess the prevalence of a Wolbachia lineage (wPpe) in 32 field populations of Pratylenchus penetrans. Populations were examined from eight different plant species in Washington, Oregon, and California. Nematodes were also screened for the endosymbiotic bacterium Cardinium (cPpe) that was recently shown to co-infect P. penetrans. Results identified wPpe in 9/32 and cPpe in 1/32 of P. penetrans field populations analyzed. No co-infection was observed in field populations. Wolbachia was detected in nematodes from 4/8 plant-hosts examined (raspberry, strawberry, clover, and lily), and in all three states surveyed. Cardinium was detected in nematodes from mint in Washington. In the wPpe-infected P. penetrans populations collected from raspberry, the prevalence of wPpe infection ranged from 11 to 58%. This pattern is unlike that in filarial nematodes where Wolbachia is an obligate mutualist and occurs in 100% of the host. Further analysis of wPpe-infected populations revealed female-skewed sex ratios (up to 96%), with the degree of skew positively correlating with wPpe prevalence. Uninfected nematode populations had approximately equal numbers of males and females. Comparisons of 54 wPpe 16S ribosomal RNA sequences revealed high similarity across the geographic isolates, with 45 of 54 isolates being identical at this locus. The complete absence of wPpe among some populations and low prevalence in others suggest that this endosymbiont is not an obligate mutualist of P. penetrans. The observed sex ratio bias in wPpe-infected nematode populations is similar to that observed in arthropods where Wolbachia acts as a reproductive manipulator, raising the question of a similar role in plant-parasitic nematodes.}, } @article {pmid31128084, year = {2019}, author = {Guo, Y and Gong, JT and Mo, PW and Huang, HJ and Hong, XY}, title = {Wolbachia localization during Laodelphax striatellus embryogenesis.}, journal = {Journal of insect physiology}, volume = {116}, number = {}, pages = {125-133}, doi = {10.1016/j.jinsphys.2019.05.006}, pmid = {31128084}, issn = {1879-1611}, mesh = {Animals ; *Embryonic Development ; Hemiptera/embryology/*microbiology ; Phylogeny ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachia are intracellular bacteria carried by thousands of arthropod species. The success of Wolbachia is due to efficient vertical transmission by the host maternal germline. Wolbachia's behavior during host oogenesis is well characterized, although their behavior during embryogenesis is unclear. Vertical transmission of Wolbachia wStri in the small brown planthopper, Laodelphax striatellus is extraordinarily efficient. To understand why, we investigated its localization and dynamics in L. striatellus embryos. Microscopic observations indicated that the Wolbachia were mainly localized at the anterior region of the embryo during early embryogenesis. The distribution of Wolbachia within the anterior region was established during oogenesis, and according to a phylogenetic analysis, may be due to intrinsic factors in Wolbachia. We observed that wStri migrated to the posterior part cells during late embryogenesis, in the region where gonads were formed. An expression profile of Wolbachia-infected host embryonic development genes revealed Ddx1 mRNAs, which is required for host viability and in the germ line, accumulated in the posterior region of 3-day-old embryos, while other development genes mRNAs were significantly more abundant in the posterior region of 6-day-old embryos. These genes thus appear to be associated with the localization of Wolbachia wStri in the anterior region, although their functions remain unclear. These results can explain Wolbachia wStri high prevalence in L. striatellus.}, } @article {pmid31121066, year = {2019}, author = {Hubert, J and Nesvorna, M and Klimov, P and Dowd, SE and Sopko, B and Erban, T}, title = {Differential allergen expression in three Tyrophagus putrescentiae strains inhabited by distinct microbiome.}, journal = {Allergy}, volume = {74}, number = {12}, pages = {2502-2507}, doi = {10.1111/all.13921}, pmid = {31121066}, issn = {1398-9995}, support = {18-04-01092//Russian Foundation for Basic Research/International ; 19-09998S//Czech Science Foundation (GACR)/International ; 16-14-10109//Russian Science Foundation/International ; CZ.02.2.69/0.0/0.0/16_027/0008503//Ministry of Education, Youth and Sports of the Czech Republic/International ; RO0418//Ministry of Agriculture of the Czech Republic/International ; }, mesh = {Acaridae/*genetics/*immunology ; Allergens/*genetics/*immunology ; Animals ; *Gene Expression ; Gene Expression Profiling ; *Microbiota ; Transcriptome ; }, } @article {pmid31120156, year = {2020}, author = {Chouin-Carneiro, T and Ant, TH and Herd, C and Louis, F and Failloux, AB and Sinkins, SP}, title = {Wolbachia strain wAlbA blocks Zika virus transmission in Aedes aegypti.}, journal = {Medical and veterinary entomology}, volume = {34}, number = {1}, pages = {116-119}, pmid = {31120156}, issn = {1365-2915}, support = {202888/WT_/Wellcome Trust/United Kingdom ; ZK/16-021/MRC_/Medical Research Council/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Female ; Mosquito Vectors/microbiology/virology ; Wolbachia/*physiology ; Zika Virus/physiology ; Zika Virus Infection/*transmission ; }, abstract = {Transinfections of the maternally transmitted endosymbiont Wolbachia pipientis can reduce RNA virus replication and prevent transmission by Aedes aegypti, and also have the capacity to invade wild-type populations, potentially reaching and maintaining high infection frequencies. Levels of virus transmission blocking are positively correlated with Wolbachia intracellular density. Despite reaching high densities in Ae. aegypti, transinfections of wAlbA, a strain native to Aedes albopictus, showed no blocking of Semliki Forest Virus in previous intrathoracic injection challenges. To further characterize wAlbA blocking in Ae. aegypti, adult females were intrathoracically challenged with Zika (ZIKV) and dengue viruses, and then fed a ZIKV-containing bloodmeal. No blocking was observed with either virus when challenged by intrathoracic injection. However, when ZIKV was delivered orally, wAlbA-infected females showed a significant reduction in viral replication and dissemination compared with uninfected controls, as well as a complete absence of virus in saliva. Although other Wolbachia strains have been shown to cause more robust viral blocking in Ae. aegypti, these findings demonstrate that, in principle, wAlbA could be used to reduce virus transmission in this species. Moreover, the results highlight the potential for underestimation of the strength of virus-blocking when based on intrathoracic injection compared with more natural oral challenges.}, } @article {pmid31115477, year = {2019}, author = {Gómez-Díaz, JS and Montoya-Lerma, J and Muñoz Valencia, V}, title = {Prevalence and Diversity of Endosymbionts in Cassava Whiteflies (Hemiptera: Aleyrodidae) From Colombia.}, journal = {Journal of insect science (Online)}, volume = {19}, number = {3}, pages = {}, pmid = {31115477}, issn = {1536-2442}, mesh = {Animals ; Hemiptera/genetics/*microbiology ; Manihot ; Nymph/microbiology ; Phylogeny ; Symbiosis ; }, abstract = {Whiteflies cause huge economic losses for cassava (Manihot esculenta Crantz) cultivation. Damage can be caused directly when the insects feed on the phloem and/or indirectly by the transmission of viruses. It has been found that whiteflies maintain a close relationship with some endosymbiotic bacteria and that this interaction produces different effects on host biology and can also facilitate viral transmission. This study aimed to characterize the diversity of secondary endosymbionts (SE) present in whiteflies associated with cassava. Whitefly adults and nymphs were collected from cassava crops at nine locations in Southwestern Colombia. Molecular identification of insects and endosymbionts was carried out using specific mtCOI, wsp, 23s rRNA, and 16s rRNA primers. Phylogenetic trees were constructed from these sequences, both for whitefly species and the endosymbionts found. In addition, morphological identification of whitefly species was made using last instar nymphs. Molecular and morphological evaluation revealed that the most abundant whitefly species was Trialeurodes variabilis (Quaintance) followed by Aleurotrachelus socialis Bondar and Bemisia tuberculata Bondar. One hundred percent of the individuals contained the primary endosymbiont Portiera. The SE Rickettsia, Hamiltonella, Wolbachia, and Fritschea were not detected in the samples tested. Prevalence of Cardinium and Arsenophonus were variable at each locality, Cardinium being most prevalent in A. socialis adults. This study is the first report on the presence of Cardinium and Arsenophonus in A. socialis and T. variabilis. It is also the first report of endosymbiotic diversity in whiteflies associated with cassava in Colombia.}, } @article {pmid31107912, year = {2019}, author = {Caragata, EP and Rocha, MN and Pereira, TN and Mansur, SB and Dutra, HLC and Moreira, LA}, title = {Pathogen blocking in Wolbachia-infected Aedes aegypti is not affected by Zika and dengue virus co-infection.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {5}, pages = {e0007443}, pmid = {31107912}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*virology ; Animals ; Brazil ; Dengue Virus/genetics/*physiology ; Female ; Male ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology/*virology ; Wolbachia/genetics/*physiology ; Zika Virus/genetics/*physiology ; }, abstract = {BACKGROUND: Wolbachia's ability to restrict arbovirus transmission makes it a promising tool to combat mosquito-transmitted diseases. Wolbachia-infected Aedes aegypti are currently being released in locations such as Brazil, which regularly experience concurrent outbreaks of different arboviruses. A. aegypti can become co-infected with, and transmit multiple arboviruses with one bite, which can complicate patient diagnosis and treatment.

Using experimental oral infection of A. aegypti and then RT-qPCR, we examined ZIKV/DENV-1 and ZIKV/DENV-3 co-infection in Wolbachia-infected A. aegypti and observed that Wolbachia-infected mosquitoes experienced lower prevalence of infection and viral load than wildtype mosquitoes, even with an extra infecting virus. Critically, ZIKV/DENV co-infection had no significant impact on Wolbachia's ability to reduce viral transmission. Wolbachia infection also strongly altered expression levels of key immune genes Defensin C and Transferrin 1, in a virus-dependent manner.

CONCLUSIONS/SIGNIFICANCE: Our results suggest that pathogen interference in Wolbachia-infected A. aegypti is not adversely affected by ZIKV/DENV co-infection, which suggests that Wolbachia-infected A. aegypti will likely prove suitable for controlling mosquito-borne diseases in environments with complex patterns of arbovirus transmission.}, } @article {pmid31100967, year = {2019}, author = {Rahayu, A and Saraswati, U and Supriyati, E and Kumalawati, DA and Hermantara, R and Rovik, A and Daniwijaya, EW and Fitriana, I and Setyawan, S and Ahmad, RA and Wardana, DS and Indriani, C and Utarini, A and Tantowijoyo, W and Arguni, E}, title = {Prevalence and Distribution of Dengue Virus in Aedes aegypti in Yogyakarta City before Deployment of Wolbachia Infected Aedes aegypti.}, journal = {International journal of environmental research and public health}, volume = {16}, number = {10}, pages = {}, pmid = {31100967}, issn = {1660-4601}, mesh = {Aedes/*microbiology ; Animals ; *Biological Control Agents ; Cities ; Dengue Virus/*isolation & purification ; Female ; Indonesia ; *Mosquito Vectors ; Seasons ; Serogroup ; *Wolbachia ; }, abstract = {Indonesia is one of the countries where dengue infection is prevalent. In this study we measure the prevalence and distribution of dengue virus (DENV) DENV-infected Aedes aegypti in Yogyakarta City, Indonesia, during the wet season when high dengue transmission period occurred, as baseline data before implementation of a Wolbachia-infected Aedes aegypti trial for dengue control. We applied One-Step Multiplex Real Time PCR (RT-PCR) for the type-specific-detection of dengue viruses in field-caught adult Aedes aegypti mosquitoes. In a prospective field study conducted from December 2015 to May 2016, adult female Aedes aegypti were caught from selected areas in Yogyakarta City, and then screened by using RT-PCR. During the survey period, 36 (0.12%) mosquitoes from amongst 29,252 female mosquitoes were positive for a DENV type. In total, 22.20% of dengue-positive mosquitoes were DENV-1, 25% were DENV-2, 17% were DENV-3, but none were positive for DENV-4. This study has provided dengue virus infection prevalence in field-caught Aedes aegypti and its circulating serotype in Yogyakarta City before deployment of Wolbachia-infected Aedes aegypti.}, } @article {pmid31095639, year = {2019}, author = {Hosseinzadeh, S and Ramsey, J and Mann, M and Bennett, L and Hunter, WB and Shams-Bakhsh, M and Hall, DG and Heck, M}, title = {Color morphology of Diaphorina citri influences interactions with its bacterial endosymbionts and 'Candidatus Liberibacter asiaticus'.}, journal = {PloS one}, volume = {14}, number = {5}, pages = {e0216599}, pmid = {31095639}, issn = {1932-6203}, mesh = {Animals ; Citrus/*microbiology/parasitology ; Color ; Hemiptera/*microbiology/physiology ; Hemocyanins/metabolism ; *Host-Pathogen Interactions ; Insect Vectors/*microbiology/physiology ; Plant Diseases/*microbiology ; Rhizobiaceae/isolation & purification/*physiology ; *Symbiosis ; }, abstract = {Diaphorina citri is a vector of 'Candidatus Liberibacter asiaticus,' (CLas), associated with Huanglongbing, (HLB, or citrus greening) disease in citrus. D. citri exhibits three different color morph variants, blue, gray and yellow. Blue morphs have a greater capacity for long-distance flight as compared to non-blue morphs, but little else is known about how color morphology influences vector characteristics. In this study, we show that the color morphology of the insect is derived from pigmented cells of the fat body. Blue morphs acquire a lower level of CLas in their bodies from infected trees as compared to their gray and yellow conspecifics, referred to in this paper collectively as non-blue morphs. Accordingly, CLas titer in citrus leaves inoculated by non-blue insects was 6-fold higher than in leaves inoculated by blue insects. Blue color morphs harbored lower titers of Wolbachia and 'Candidatus Profftella armatura,' two of the D. citri bacterial endosymbionts. Expression of hemocyanin, a copper-binding oxygen transport protein responsible for the blue coloration of hemolymph of other arthropods and mollusks, was previously correlated with blue color morphology and is highly up-regulated in insects continuously reared on CLas infected citrus trees. Based on our results, we hypothesized that a reduction of hemocyanin expression would reduce the D. citri immune response and an increase in the titer of CLas would be observed. Surprisingly, a specific 3-fold reduction of hemocyanin-1 transcript levels using RNA silencing in blue adult D. citri morphs had an approximately 2-fold reduction on the titer of CLas. These results suggest that hemocyanin signaling from the fat body may have multiple functions in the regulation of bacterial titers in D. citri, and that hemocyanin is one of multiple psyllid genes involved in regulating CLas titer.}, } @article {pmid31092581, year = {2019}, author = {Teramoto, T and Huang, X and Armbruster, PA and Padmanabhan, R}, title = {Infection of Aedes albopictus Mosquito C6/36 Cells with the wMelpop Strain of Wolbachia Modulates Dengue Virus-Induced Host Cellular Transcripts and Induces Critical Sequence Alterations in the Dengue Viral Genome.}, journal = {Journal of virology}, volume = {93}, number = {15}, pages = {}, pmid = {31092581}, issn = {1098-5514}, mesh = {*Aedes ; Animals ; Cell Line ; Cercopithecus ; Cricetinae ; Dengue Virus/genetics/*growth & development ; Gene Expression Profiling ; *Gene Expression Regulation ; *Genome, Viral ; Host Microbial Interactions ; *Microbial Interactions ; *Mutation ; Sequence Analysis, RNA ; Virus Replication ; Wolbachia/*growth & development ; }, abstract = {Dengue virus (DENV) causes frequent epidemics infecting ∼390 million people annually in over 100 countries. There are no approved vaccines or antiviral drugs for treatment of infected patients. However, there is a novel approach to control DENV transmission by the mosquito vectors, Aedes aegypti and Aedes albopictus, using the Wolbachia symbiont. The wMelPop strain of Wolbachia suppresses DENV transmission and shortens the mosquito life span. However, the underlying mechanism is poorly understood. To clarify this mechanism, either naive A. albopictus (C6/36) or wMelPop-C6/36 cells were infected with DENV serotype 2 (DENV2). Analysis of host transcript profiles by transcriptome sequencing (RNAseq) revealed that the presence of wMelPop dramatically altered the mosquito host cell transcription in response to DENV2 infection. The viral RNA evolved from wMelPop-C6/36 cells contained low-frequency mutations (∼25%) within the coding region of transmembrane domain 1 (TMD1) of E protein. Mutations with >97% frequencies were distributed within other regions of E, the NS5 RNA-dependent RNA polymerase (NS5POL) domain, and the TMDs of NS2A, NS2B, and NS4B. Moreover, while DENV2-infected naive C6/36 cells showed syncytium formation, DENV2-infected wMelPop-C6/36 cells did not. The Wolbachia-induced mutant DENV2 can readily infect and replicate in naive C6/36 cells, whereas in mutant DENV2-infected BHK-21 or Vero cells, virus replication was delayed. In LLC-MK2 cells, the mutant failed to produce plaques. Additionally, in BHK-21 cells, many mutations in the viral genome reverted to the wild type (WT) and compensatory mutations in NS3 gene appeared. Our results indicate that wMelPop impacts significantly the interactions of DENV2 with mosquito and mammalian host cells.IMPORTANCE Mosquito-borne diseases are of global significance causing considerable morbidity and mortality throughout the world. Dengue virus (DENV; serotypes 1 to 4), a member of the Flavivirus genus of the Flaviviridae family, causes millions of infections annually. Development of a safe vaccine is hampered due to absence of cross-protection and increased risk in secondary infections due to antibody-mediated immune enhancement. Infection of vector mosquitoes with Wolbachia bacteria offers a novel countermeasure to suppress DENV transmission, but the mechanisms are poorly understood. In this study, the host transcription profiles and viral RNA sequences were analyzed in naive A. albopictus (C6/36) and wMelPop-C6/36 cells by RNAseq. Our results showed that the wMelPop symbiont caused profound changes in host transcription profiles and morphology of DENV2-infected C6/36 cells. Accumulation of several mutations throughout DENV2 RNA resulted in loss of infectivity of progeny virions. Our findings offer new insights into the mechanism of Wolbachia-mediated suppression of DENV transmission.}, } @article {pmid31088494, year = {2019}, author = {Shah, A and Hoffman, JI and Schielzeth, H}, title = {Transcriptome assembly for a colour-polymorphic grasshopper (Gomphocerus sibiricus) with a very large genome size.}, journal = {BMC genomics}, volume = {20}, number = {1}, pages = {370}, pmid = {31088494}, issn = {1471-2164}, support = {SCHI 1188/1-1//Deutsche Forschungsgemeinschaft/ ; }, mesh = {Animals ; Contig Mapping ; Female ; Gene Expression Profiling/*methods ; Genetic Association Studies ; Genome Size ; Grasshoppers/*genetics ; High-Throughput Nucleotide Sequencing ; Male ; Mitochondria/*genetics ; Molecular Sequence Annotation ; Sequence Analysis, RNA/*methods ; }, abstract = {BACKGROUND: The club-legged grasshopper Gomphocerus sibiricus is a Gomphocerinae grasshopper with a promising future as model species for studying the maintenance of colour-polymorphism, the genetics of sexual ornamentation and genome size evolution. However, limited molecular resources are available for this species. Here, we present a de novo transcriptome assembly as reference resource for gene expression studies. We used high-throughput Illumina sequencing to generate 5,070,036 paired-end reads after quality filtering. We then combined the best-assembled contigs from three different de novo transcriptome assemblers (Trinity, SOAPdenovo-trans and Oases/Velvet) into a single assembly.

RESULTS: This resulted in 82,251 contigs with a N50 of 1357 and a TransRate assembly score of 0.325, which compares favourably with other orthopteran transcriptome assemblies. Around 87% of the transcripts could be annotated using InterProScan 5, BLASTx and the dammit! annotation pipeline. We identified a number of genes involved in pigmentation and green pigment metabolism pathways. Furthermore, we identified 76,221 putative single nucleotide polymorphisms residing in 8400 contigs. We also assembled the mitochondrial genome and investigated levels of sequence divergence with other species from the genus Gomphocerus. Finally, we detected and assembled Wolbachia sequences, which revealed close sequence similarity to the strain pel wPip.

CONCLUSIONS: Our study has generated a significant resource for uncovering genotype-phenotype associations in a species with an extraordinarily large genome, while also providing mitochondrial and Wolbachia sequences that will be useful for comparative studies.}, } @article {pmid31083952, year = {2019}, author = {Fabre, B and Korona, D and Lees, JG and Lazar, I and Livneh, I and Brunet, M and Orengo, CA and Russell, S and Lilley, KS}, title = {Comparison of Drosophila melanogaster Embryo and Adult Proteome by SWATH-MS Reveals Differential Regulation of Protein Synthesis, Degradation Machinery, and Metabolism Modules.}, journal = {Journal of proteome research}, volume = {18}, number = {6}, pages = {2525-2534}, doi = {10.1021/acs.jproteome.9b00076}, pmid = {31083952}, issn = {1535-3907}, support = {BB/L002817/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/growth & development/metabolism/microbiology ; Embryo, Nonmammalian/metabolism/microbiology ; Gene Expression Regulation, Developmental/genetics ; Protein Biosynthesis/*genetics ; Proteolysis ; Proteome/*genetics/metabolism ; Proteomics/methods ; Transcriptome/*genetics ; Wolbachia/pathogenicity ; }, abstract = {An important area of modern biology consists of understanding the relationship between genotype and phenotype. However, to understand this relationship it is essential to investigate one of the principal links between them: the proteome. With the development of recent mass-spectrometry approaches, it is now possible to quantify entire proteomes and thus relate them to different phenotypes. Here, we present a comparison of the proteome of two extreme developmental states in the well-established model organism Drosophila melanogaster: adult and embryo. Protein modules such as ribosome, proteasome, tricarboxylic acid cycle, glycolysis, or oxidative phosphorylation were found differentially expressed between the two developmental stages. Analysis of post-translation modifications of the proteins identified in this study indicates that they generally follow the same trend as their corresponding protein. Comparison between changes in the proteome and the transcriptome highlighted patterns of post-transcriptional regulation for the subunits of protein complexes such as the ribosome and the proteasome, whereas protein from modules such as TCA cycle, glycolysis, and oxidative phosphorylation seem to be coregulated at the transcriptional level. Finally, the impact of the endosymbiont Wolbachia pipientis on the proteome of both developmental states was also investigated.}, } @article {pmid31074963, year = {2019}, author = {Xu, Z and Fang, SM and Bakowski, MA and Rateb, ME and Yang, D and Zhu, X and Huang, Y and Zhao, LX and Jiang, Y and Duan, Y and Hull, MV and McNamara, CW and Shen, B}, title = {Discovery of Kirromycins with Anti- Wolbachia Activity from Streptomyces sp. CB00686.}, journal = {ACS chemical biology}, volume = {14}, number = {6}, pages = {1174-1182}, doi = {10.1021/acschembio.9b00086}, pmid = {31074963}, issn = {1554-8937}, mesh = {Animals ; Anti-Bacterial Agents/*chemistry/*pharmacology ; Biological Products/chemistry/pharmacology ; Drosophila/microbiology ; *Drug Discovery ; HEK293 Cells ; Humans ; Pyridones/chemistry/pharmacology ; Streptomyces/*chemistry ; Wolbachia/*drug effects ; }, abstract = {Lymphatic filariasis and onchocerciasis diseases caused by filarial parasite infections can lead to profound disability and affect millions of people worldwide. Standard mass drug administration campaigns require repetitive delivery of anthelmintics for years to temporarily block parasite transmission but do not cure infection because long-lived adult worms survive the treatment. Depletion of the endosymbiont Wolbachia, present in most filarial nematode species, results in death of adult worms and therefore represents a promising target for the treatment of filariasis. Here, we used a high-content imaging assay to screen the pure compounds collection of the natural products library at The Scripps Research Institute for anti- Wolbachia activity, leading to the identification of kirromycin B (1) as a lead candidate. Two additional congeners, kirromycin (2) and kirromycin C (3), were isolated and characterized from the same producing strain Streptomyces sp. CB00686. All three kirromycin congeners depleted Wolbachia in LDW1 Drosophila cells in vitro with half-maximal inhibitory concentrations (IC50) in nanomolar range, while doxycycline, a registered drug with anti- Wolbachia activity, showed lower activity with an IC50 of 152 ± 55 nM. Furthermore, 1-3 eliminated the Wolbachia endosymbiont in Brugia pahangi ovaries ex vivo with higher efficiency (65%-90%) at 1 μM than that of doxycycline (50%). No cytotoxicity against HEK293T and HepG2 mammalian cells was observed with 1-3 at the highest concentration (40 μM) used in the assay. These results suggest kirromycin is an effective lead scaffold, further exploration of which could potentially lead to the development of novel treatments for filarial nematode infections.}, } @article {pmid31068442, year = {2019}, author = {Bakowski, MA and Shiroodi, RK and Liu, R and Olejniczak, J and Yang, B and Gagaring, K and Guo, H and White, PM and Chappell, L and Debec, A and Landmann, F and Dubben, B and Lenz, F and Struever, D and Ehrens, A and Frohberger, SJ and Sjoberg, H and Pionnier, N and Murphy, E and Archer, J and Steven, A and Chunda, VC and Fombad, FF and Chounna, PW and Njouendou, AJ and Metuge, HM and Ndzeshang, BL and Gandjui, NV and Akumtoh, DN and Kwenti, TDB and Woods, AK and Joseph, SB and Hull, MV and Xiong, W and Kuhen, KL and Taylor, MJ and Wanji, S and Turner, JD and Hübner, MP and Hoerauf, A and Chatterjee, AK and Roland, J and Tremblay, MS and Schultz, PG and Sullivan, W and Chu, XJ and Petrassi, HM and McNamara, CW}, title = {Discovery of short-course antiwolbachial quinazolines for elimination of filarial worm infections.}, journal = {Science translational medicine}, volume = {11}, number = {491}, pages = {}, doi = {10.1126/scitranslmed.aav3523}, pmid = {31068442}, issn = {1946-6242}, mesh = {Animals ; Anti-Bacterial Agents/chemistry/pharmacology/*therapeutic use ; Disease Models, Animal ; *Drug Discovery ; Female ; Filariasis/*drug therapy/*parasitology ; Filarioidea/drug effects/microbiology/*physiology ; High-Throughput Screening Assays ; Mice ; Phenotype ; Quinazolines/chemistry/pharmacology/*therapeutic use ; Small Molecule Libraries ; Wolbachia/drug effects ; }, abstract = {Parasitic filarial nematodes cause debilitating infections in people in resource-limited countries. A clinically validated approach to eliminating worms uses a 4- to 6-week course of doxycycline that targets Wolbachia, a bacterial endosymbiont required for worm viability and reproduction. However, the prolonged length of therapy and contraindication in children and pregnant women have slowed adoption of this treatment. Here, we describe discovery and optimization of quinazolines CBR417 and CBR490 that, with a single dose, achieve >99% elimination of Wolbachia in the in vivo Litomosoides sigmodontis filarial infection model. The efficacious quinazoline series was identified by pairing a primary cell-based high-content imaging screen with an orthogonal ex vivo validation assay to rapidly quantify Wolbachia elimination in Brugia pahangi filarial ovaries. We screened 300,368 small molecules in the primary assay and identified 288 potent and selective hits. Of 134 primary hits tested, only 23.9% were active in the worm-based validation assay, 8 of which contained a quinazoline heterocycle core. Medicinal chemistry optimization generated quinazolines with excellent pharmacokinetic profiles in mice. Potent antiwolbachial activity was confirmed in L. sigmodontis, Brugia malayi, and Onchocerca ochengi in vivo preclinical models of filarial disease and in vitro selectivity against Loa loa (a safety concern in endemic areas). The favorable efficacy and in vitro safety profiles of CBR490 and CBR417 further support these as clinical candidates for treatment of filarial infections.}, } @article {pmid31066101, year = {2019}, author = {Hodson, CN and Perlman, SJ}, title = {Population biology of a selfish sex ratio distorting element in a booklouse (Psocodea: Liposcelis).}, journal = {Journal of evolutionary biology}, volume = {32}, number = {8}, pages = {825-832}, doi = {10.1111/jeb.13484}, pmid = {31066101}, issn = {1420-9101}, mesh = {Animals ; Female ; Male ; Neoptera/*genetics ; Sex Ratio ; }, abstract = {Arthropods harbour a variety of selfish genetic elements that manipulate reproduction to be preferentially transmitted to future generations. A major ongoing question is to understand how these elements persist in nature. In this study, we examine the population dynamics of an unusual selfish sex ratio distorter in a recently discovered species of booklouse, Liposcelis sp. (Psocodea: Liposcelididae) to gain a better understanding of some of the factors that may affect the persistence of this element. Females that carry the selfish genetic element only ever produce daughters, although they are obligately sexual. These females also only transmit the maternal half of their genome. We performed a replicated population cage experiment, varying the initial frequency of females that harbour the selfish element, and following female frequencies for 20 months. The selfish genetic element persisted in all cages, often reaching very high (and thus severely female-biased) frequencies. Surprisingly, we also found that females that carry the selfish genetic element had much lower fitness than their nondistorter counterparts, with lower lifetime fecundity, slower development and a shorter egg-laying period. We suggest that differential fitness plays a role in the maintenance of the selfish genetic element in this species. We believe that the genetic system in this species, paternal genome elimination, which allows maternal control of offspring sex ratio, may also be important in the persistence of the selfish genetic element, highlighting the need to consider species with diverse ecologies and genetic systems when investigating the effects of sex ratio manipulators on host populations.}, } @article {pmid31060581, year = {2019}, author = {Farnesi, LC and Belinato, TA and Gesto, JSM and Martins, AJ and Bruno, RV and Moreira, LA}, title = {Embryonic development and egg viability of wMel-infected Aedes aegypti.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {211}, pmid = {31060581}, issn = {1756-3305}, mesh = {Aedes/embryology/*microbiology ; Animals ; Cell Survival ; Embryonic Development ; Female ; Humans ; Male ; Mosquito Vectors/*microbiology ; Ovum/*growth & development ; Wolbachia/physiology ; }, abstract = {BACKGROUND: Aedes aegypti is a major disease vector in urban habitats, involved in the transmission of dengue, chikungunya and Zika. Despite innumerous attempts to contain disease outbreaks, there are neither efficient vaccines nor definite vector control methods nowadays. In recent years, an innovative strategy to control arboviruses, which exploits the endosymbiotic bacterium Wolbachia pipientis, emerged with great expectations. The success of the method depends on many aspects, including Wolbachia's cytoplasmic incompatibility and pathogen interference phenotypes, as well as its effect on host fitness. In this work, we investigated the influence the Wolbachia strain wMel exerts on embryo development and egg viability and speculate on its field release use.

METHODS: Wild-type (Br or Rockefeller) and Wolbachia-harboring specimens (wMelBr) were blood-fed and submitted to synchronous egg laying for embryo development assays. Samples were analyzed for morphological markers, developmental endpoint and egg resistance to desiccation (ERD). Quiescent egg viability over time was also assessed.

RESULTS: wMelBr samples completed embryogenesis 2-3 hours later than wild-type. This delay was also observed through the onset of both morphological and physiological markers, respectively by the moments of germband extension and ERD acquisition. Following the end of embryonic development, wMelBr eggs were slightly less resistant to desiccation and showed reduced viability levels, which rapidly decayed after 40 days into quiescence, from approximately 75% to virtually 0% in less than a month.

CONCLUSIONS: Our data revealed that the wMel strain of Wolbachia slightly delays embryogenesis and also affects egg quality, both through reduced viability and desiccation resistance. These findings suggest that, although embryonic fitness is somehow compromised by wMel infection, an efficient host reproductive manipulation through cytoplasmic incompatibility seems sufficient to overcome these effects in nature and promote bacterial invasion, as shown by successful ongoing field implementation.}, } @article {pmid31052498, year = {2019}, author = {Cao, LJ and Jiang, W and Hoffmann, AA}, title = {Life History Effects Linked to an Advantage for wAu Wolbachia in Drosophila.}, journal = {Insects}, volume = {10}, number = {5}, pages = {}, pmid = {31052498}, issn = {2075-4450}, support = {DP120100916//Australian Research Council/ ; R01 GM104325/GM/NIGMS NIH HHS/United States ; 1118640//National Health and Medical Research Council/ ; GJHZ2017-5//Beijing Academy of Agriculture and Forestry Sciences/ ; LE150100083//Australian Research Council/ ; 5R01GM104325//National Institutes of Health/ ; }, abstract = {Wolbachia endosymbiont infections can persist and spread in insect populations without causing apparent effects on reproduction of their insect hosts, but the mechanisms involved are largely unknown. Here, we test for fitness effects of the wAu infection of Drosophila simulans by comparing multiple infected and uninfected polymorphic isofemale lines derived from nature. We show a fitness advantage (higher offspring number) for lines with the wAu Wolbachia infection when breeding on grapes, but only where there was Talaromyces and Penicillium fungal mycelial growth. When breeding on laboratory medium, the wAu infection extended the development time and resulted in larger females with higher fecundity, life history traits, which may increase fitness. A chemical associated with the fungi (ochratoxin A) did not specifically alter the fitness of wAu-infected larvae, which developed slower and emerged with a greater weight regardless of toxin levels. These findings suggest that the fitness benefits of Wolbachia in natural populations may reflect life history changes that are advantageous under particular circumstances, such as when breeding occurs in rotting fruit covered by abundant mycelial growth.}, } @article {pmid31048806, year = {2019}, author = {Crunkhorn, S}, title = {Inhibiting Wolbachia to treat parasitic diseases.}, journal = {Nature reviews. Drug discovery}, volume = {}, number = {}, pages = {}, doi = {10.1038/d41573-019-00058-z}, pmid = {31048806}, issn = {1474-1784}, } @article {pmid31031440, year = {2019}, author = {Kaushik, S and Sharma, KK and Ramani, R and Lakhanpaul, S}, title = {Detection of Wolbachia Phage (WO) in Indian Lac Insect [Kerria lacca (Kerr.)] and Its Implications.}, journal = {Indian journal of microbiology}, volume = {59}, number = {2}, pages = {237-240}, pmid = {31031440}, issn = {0046-8991}, abstract = {Wolbachia, a maternally inherited bacterium induces reproductive alterations in its hosts such as feminization of males, male killing and parthenogenesis. It is the most diverse endosymbiont infecting more than 70% of the insects ranging from pests to pollinators. Kerria lacca-a hemipteran is a sedentary, oriental insect known to produce lac-the only resin of animal origin. The present study was conducted to screen the presence of Wolbachia and its associated phages in the two infrasubspecific forms (four insect lines) of K. lacca viz. kusmi and rengeeni differing from each other on the basis of host preference. Wolbachia and its associated phage were found to be prevalent in all the insect lines analyzed. We, hereby, report the presence of WO-phage (Wolbachia phage) for the first time in K. lacca. Further, phylogenetic data differentiated the kusmi and rengeeni infrasubspecific forms into two different groups on the basis of WO-phage sequences.}, } @article {pmid31028710, year = {2019}, author = {}, title = {Evolutionary Genetics of Cytoplasmic Incompatibility Genes cifA and cifB in Prophage WO of Wolbachia.}, journal = {Genome biology and evolution}, volume = {11}, number = {4}, pages = {1320}, doi = {10.1093/gbe/evz079}, pmid = {31028710}, issn = {1759-6653}, } @article {pmid31024463, year = {2019}, author = {Monteiro, VVS and Navegantes-Lima, KC and de Lemos, AB and da Silva, GL and de Souza Gomes, R and Reis, JF and Rodrigues Junior, LC and da Silva, OS and Romão, PRT and Monteiro, MC}, title = {Aedes-Chikungunya Virus Interaction: Key Role of Vector Midguts Microbiota and Its Saliva in the Host Infection.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {492}, pmid = {31024463}, issn = {1664-302X}, abstract = {Aedes mosquitoes are important vectors for emerging diseases caused by arboviruses, such as chikungunya (CHIKV). These viruses' main transmitting species are Aedes aegypti and Ae. albopictus, which are present in tropical and temperate climatic areas all over the globe. Knowledge of vector characteristics is fundamentally important to the understanding of virus transmission. Only female mosquitoes are able to transmit CHIKV to the vertebrate host since they are hematophagous. In addition, mosquito microbiota is fundamentally important to virus infection in the mosquito. Microorganisms are able to modulate viral transmission in the mosquito, such as bacteria of the Wolbachia genus, which are capable of preventing viral infection, or protozoans of the Ascogregarina species, which are capable of facilitating virus transmission between mosquitoes and larvae. The competence of the mosquito is also important in the transmission of the virus to the vertebrate host, since their saliva has several substances with biological effects, such as immunomodulators and anticoagulants, which are able to modulate the host's response to the virus, interfering in its pathogenicity and virulence. Understanding the Aedes vector-chikungunya interaction is fundamentally important since it can enable the search for new methods of combating the virus' transmission.}, } @article {pmid31008514, year = {2019}, author = {Mains, JW and Kelly, PH and Dobson, KL and Petrie, WD and Dobson, SL}, title = {Localized Control of Aedes aegypti (Diptera: Culicidae) in Miami, FL, via Inundative Releases of Wolbachia-Infected Male Mosquitoes.}, journal = {Journal of medical entomology}, volume = {56}, number = {5}, pages = {1296-1303}, doi = {10.1093/jme/tjz051}, pmid = {31008514}, issn = {1938-2928}, mesh = {*Aedes/microbiology ; Animals ; Florida ; Male ; *Mosquito Control ; *Pest Control, Biological ; Population Dynamics ; Wolbachia/*physiology ; }, abstract = {As part of the response to autochthonous Zika transmission in the United States, the City of South Miami implemented a 6-mo period in which Wolbachia-infected WB1 Aedes aegypti (L.) males were released into an ~170-acre area. Intracellular Wolbachia bacteria infections in Ae. aegypti cause early embryonic arrest (known as cytoplasmic incompatibility [CI]) and egg hatch failure, and inundative introductions have been suggested as a potential control tool. Throughout the release period, the Ae. aegypti population was monitored within both the release area and an equivalent area that did not receive WB1 male releases. The results show a significant reduction in egg hatch at the area receiving WB1 males, which is consistent with expectations for CI. Similarly, the number of Ae. aegypti was significantly reduced at the area receiving WB1 males, relative to the untreated area. The observed population reduction and results encourage additional work and replication of the Wolbachia biopesticide approach against Ae. aegypti, as an additional tool to be integrated with existing control tools for the control of this medically important vector and nuisance pest.}, } @article {pmid31006611, year = {2019}, author = {Vila, A and Estrada-Peña, A and Altet, L and Cusco, A and Dandreano, S and Francino, O and Halos, L and Roura, X}, title = {Endosymbionts carried by ticks feeding on dogs in Spain.}, journal = {Ticks and tick-borne diseases}, volume = {10}, number = {4}, pages = {848-852}, doi = {10.1016/j.ttbdis.2019.04.003}, pmid = {31006611}, issn = {1877-9603}, mesh = {Animals ; Bacteria/genetics/isolation & purification/*pathogenicity ; DNA, Bacterial/genetics ; Dermacentor/microbiology ; Dogs/*parasitology ; Feeding Behavior ; Female ; Ixodidae/microbiology ; Male ; Polymerase Chain Reaction ; Rhipicephalus sanguineus/microbiology ; Spain ; *Symbiosis ; Tick Infestations/microbiology/*veterinary ; Tick-Borne Diseases/microbiology/*veterinary ; Ticks/*microbiology ; }, abstract = {Studies on tick microbial communities historically focused on tick-borne pathogens. However, there is an increasing interest in capturing relationships among non-pathogenic endosymbionts and exploring their relevance for tick biology. The present study included a total of 1600 adult ticks collected from domestic dogs in 4 different biogeographical regions of Spain. Each pool formed by 1 to 10 halves of individuals representing one specific ticks species was examined by PCR for the presence of Coxiellaceae, Rickettsia spp., Rickettsiales, Wolbachia spp., and other bacterial DNA. Of the pools analyzed, 92% tested positive for endosymbiont-derived DNA. Coxiella spp. endosymbionts were the most prevalent microorganisms, being always present in Rhipicephalus sanguineus sensu lato (s.l.) pools. Rickettsia spp. DNA was detected in 60% of Dermacentor reticulatus pools and 40% of R. sanguineus s.l. pools, with a higher diversity of Rickettsia species in R. sanguineus s.l. pools. Our study reveals a negative relationship of Rickettsia massiliae with the presence of tick-borne pathogens in the same pool of ticks. An additional endosymbiont, 'Candidatus Rickettsiella isopodorum', was only detected in D. reticulatus pools. Data from this study indicate that dogs in Spain are exposed to several endosymbionts. Due to the importance of tick-borne pathogens, characterizing the role of endosymbionts for tick physiology and prevalence, may lead to novel control strategies.}, } @article {pmid31003827, year = {2019}, author = {Shropshire, JD and Leigh, B and Bordenstein, SR and Duplouy, A and Riegler, M and Brownlie, JC and Bordenstein, SR}, title = {Models and Nomenclature for Cytoplasmic Incompatibility: Caution over Premature Conclusions - A Response to Beckmann et al.}, journal = {Trends in genetics : TIG}, volume = {35}, number = {6}, pages = {397-399}, doi = {10.1016/j.tig.2019.03.004}, pmid = {31003827}, issn = {0168-9525}, support = {F32 AI140694/AI/NIAID NIH HHS/United States ; }, mesh = {Cytoplasm ; *Genetic Fitness ; *Models, Biological ; Operon ; *Terminology as Topic ; Toxin-Antitoxin Systems/genetics ; Virulence Factors/genetics ; }, } @article {pmid31002720, year = {2019}, author = {Ross, PA and Ritchie, SA and Axford, JK and Hoffmann, AA}, title = {Loss of cytoplasmic incompatibility in Wolbachia-infected Aedes aegypti under field conditions.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {4}, pages = {e0007357}, pmid = {31002720}, issn = {1935-2735}, support = {108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; Environment ; Female ; *Hot Temperature ; Insect Vectors/microbiology ; Male ; Pest Control, Biological/*methods ; Queensland ; Wolbachia/*growth & development ; }, abstract = {Wolbachia bacteria are now being introduced into Aedes aegypti mosquito populations for dengue control. When Wolbachia infections are at a high frequency, they influence the local transmission of dengue by direct virus blocking as well as deleterious effects on vector mosquito populations. However, the effectiveness of this strategy could be influenced by environmental temperatures that decrease Wolbachia density, thereby reducing the ability of Wolbachia to invade and persist in the population and block viruses. We reared wMel-infected Ae. aegypti larvae in the field during the wet season in Cairns, North Queensland. Containers placed in the shade produced mosquitoes with a high Wolbachia density and little impact on cytoplasmic incompatibility. However, in 50% shade where temperatures reached 39°C during the day, wMel-infected males partially lost their ability to induce cytoplasmic incompatibility and females had greatly reduced egg hatch when crossed to infected males. In a second experiment under somewhat hotter conditions (>40°C in 50% shade), field-reared wMel-infected females had their egg hatch reduced to 25% when crossed to field-reared wMel-infected males. Wolbachia density was reduced in 50% shade for both sexes in both experiments, with some mosquitoes cleared of their Wolbachia infections entirely. To investigate the critical temperature range for the loss of Wolbachia infections, we held Ae. aegypti eggs in thermocyclers for one week at a range of cyclical temperatures. Adult wMel density declined when eggs were held at 26-36°C or above with complete loss at 30-40°C, while the density of wAlbB remained high until temperatures were lethal. These findings suggest that high temperature effects on Wolbachia are potentially substantial when breeding containers are exposed to partial sunlight but not shade. Heat stress could reduce the ability of Wolbachia infections to invade mosquito populations in some locations and may compromise the ability of Wolbachia to block virus transmission in the field. Temperature effects may also have an ecological impact on mosquito populations given that a proportion of the population becomes self-incompatible.}, } @article {pmid31001816, year = {2019}, author = {Meany, MK and Conner, WR and Richter, SV and Bailey, JA and Turelli, M and Cooper, BS}, title = {Loss of cytoplasmic incompatibility and minimal fecundity effects explain relatively low Wolbachia frequencies in Drosophila mauritiana.}, journal = {Evolution; international journal of organic evolution}, volume = {73}, number = {6}, pages = {1278-1295}, pmid = {31001816}, issn = {1558-5646}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Cytoplasm/microbiology ; Drosophila/*microbiology/*physiology ; Female ; Fertility ; Islands ; Mauritius ; Phylogeny ; Wolbachia/genetics/*physiology ; }, abstract = {Maternally transmitted Wolbachia bacteria infect about half of all insect species. Many Wolbachia cause cytoplasmic incompatibility (CI) and reduced egg hatch when uninfected females mate with infected males. Although CI produces a frequency-dependent fitness advantage that leads to high equilibrium Wolbachia frequencies, it does not aid Wolbachia spread from low frequencies. Indeed, the fitness advantages that produce initial Wolbachia spread and maintain non-CI Wolbachia remain elusive. wMau Wolbachia infecting Drosophila mauritiana do not cause CI, despite being very similar to CI-causing wNo from Drosophila simulans (0.068% sequence divergence over 682,494 bp), suggesting recent CI loss. Using draft wMau genomes, we identify a deletion in a CI-associated gene, consistent with theory predicting that selection within host lineages does not act to increase or maintain CI. In the laboratory, wMau shows near-perfect maternal transmission; but we find no significant effect on host fecundity, in contrast to published data. Intermediate wMau frequencies on the island of Mauritius are consistent with a balance between unidentified small, positive fitness effects and imperfect maternal transmission. Our phylogenomic analyses suggest that group-B Wolbachia, including wMau and wPip, diverged from group-A Wolbachia, such as wMel and wRi, 6-46 million years ago, more recently than previously estimated.}, } @article {pmid31001690, year = {2019}, author = {Michalik, K and Szklarzewicz, T and Kalandyk-Kołodziejczyk, M and Michalik, A}, title = {Bacterial associates of Orthezia urticae, Matsucoccus pini, and Steingelia gorodetskia - scale insects of archaeoccoid families Ortheziidae, Matsucoccidae, and Steingeliidae (Hemiptera, Coccomorpha).}, journal = {Protoplasma}, volume = {256}, number = {5}, pages = {1205-1215}, pmid = {31001690}, issn = {1615-6102}, support = {IP2015050374//Ministerstwo Nauki i Szkolnictwa Wyższego/ ; }, mesh = {Animals ; Bacteria/*ultrastructure ; Insecta/*ultrastructure ; }, abstract = {The biological nature, ultrastructure, distribution, and mode of transmission between generations of the microorganisms associated with three species (Orthezia urticae, Matsucoccus pini, Steingelia gorodetskia) of primitive families (archaeococcoids = Orthezioidea) of scale insects were investigated by means of microscopic and molecular methods. In all the specimens of Orthezia urticae and Matsucoccus pini examined, bacteria Wolbachia were identified. In some examined specimens of O. urticae, apart from Wolbachia, bacteria Sodalis were detected. In Steingelia gorodetskia, the bacteria of the genus Sphingomonas were found. In contrast to most plant sap-sucking hemipterans, the bacterial associates of O. urticae, M. pini, and S. gorodetskia are not harbored in specialized bacteriocytes, but are dispersed in the cells of different organs. Ultrastructural observations have shown that bacteria Wolbachia in O. urticae and M. pini, Sodalis in O. urticae, and Sphingomonas in S. gorodetskia are transovarially transmitted from mother to progeny.}, } @article {pmid30999960, year = {2019}, author = {Sabūnas, V and Radzijevskaja, J and Sakalauskas, P and Petkevičius, S and Karvelienė, B and Žiliukienė, J and Lipatova, I and Paulauskas, A}, title = {Dirofilaria repens in dogs and humans in Lithuania.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {177}, pmid = {30999960}, issn = {1756-3305}, support = {VP1-3.1-ŠMM-01-V-02-003//Lietuvos Mokslo Taryba (LT)/ ; }, mesh = {Adult ; Aged ; Animals ; Child ; *Dirofilaria repens/microbiology ; Dirofilariasis/epidemiology/*parasitology ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Eye Diseases/parasitology ; Female ; Humans ; Lithuania/epidemiology ; Male ; Middle Aged ; Phylogeny ; Polymerase Chain Reaction ; Prevalence ; Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {BACKGROUND: In Lithuania, the first case of canine subcutaneous dirofilariosis was recorded in 2010. Since then, an increasing number of cases of canine dirofilariosis have been documented in different veterinary clinics throughout the country. Human dirofilariosis was diagnosed in Lithuania for the first time in September 2011. However, to the authors' knowledge, there are no published data on the presence and prevalence of autochthonous dirofilariosis in dogs and humans in the country. The present study provides information about the predominant species and prevalence of Dirofilaria in dogs and describes the cases of human dirofilariosis in Lithuania. It also outlines PCR detection of the bacterial endosymbiont Wolbachia that contributes to the inflammatory features of filarioid infection.

RESULTS: A total of 2280 blood samples and six adult worms from pet and shelter dogs were collected in the central and eastern regions of Lithuania in 2013-2015. Based on their morphological appearance, morphometric measurements and molecular analysis, all the adult nematodes were identified as Dirofilaria repens. The diagnosis of microfilariae in blood samples was based on blood smear analysis and Knott's test. The PCR and sequence analysis of the ribosomal DNA ITS2 region and cox1 gene confirmed the presence of D. repens. Overall, 61 (2.7%) of the 2280 blood samples were found to be positive for the presence of D. repens. The infection rate of D. repens was significantly higher in shelter dogs (19.0%; 19/100) than in pet dogs (1.9%; 42/2180) (χ[2] = 100.039, df = 1, P < 0.0001). Forty-nine DNA samples of D. repens-infected dogs were tested for the presence of the bacterial endosymbiont Wolbachia and, of these, 40 samples (81.6%) were found to be positive. Three ocular and six subcutaneous cases of human dirofilariosis were diagnosed in Lithuania in the period 2011-2018.

CONCLUSIONS: To the authors' knowledge, this is the first report of autochthonous D. repens infection in dogs and humans in Lithuania. The present data demonstrate that D. repens is the main etiological agent of dirofilariosis in Lithuania. The DNA of the filarioid endosymbiotic bacterium Wolbachia was detected in the vast majority of dogs infected with D. repens.}, } @article {pmid30997886, year = {2018}, author = {Carvajal, TM and Capistrano, JDR and Hashimoto, K and Go, KJD and Cruz, MAIJ and Martinez, MJLB and Tiopianco, VSP and Amalin, DM and Watanabe, K}, title = {Detection and distribution of Wolbachia endobacteria in Culex quinquefasciatus populations (Diptera : Culicidae) from Metropolitan Manila, Philippines.}, journal = {Journal of vector borne diseases}, volume = {55}, number = {4}, pages = {265-270}, doi = {10.4103/0972-9062.256561}, pmid = {30997886}, issn = {0972-9062}, mesh = {Animals ; Bacterial Proteins/genetics ; Base Sequence ; Cities ; Culex/*microbiology ; *Genetic Variation ; Mosquito Vectors/microbiology ; Philippines ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/*classification/*isolation & purification ; }, abstract = {BACKGROUND & OBJECTIVES: Culex quinquefasciatus is a peridomestic mosquito known for its ability to transmit pathogenic diseases such as filariasis and Japanese encephalitis. The development and use of novel and innovative vector control measures such as the utilization of Wolbachia, along with the existing ones, are necessary to prevent the transmission of these diseases. Studies exploring the diversity of Wolbachia, particularly in Cx. quinquefasciatus are very limited in the Philippines. Thus, the aim of the study was to detect the presence, distribution, and phylogenetic relationship of Wolbachia infections in Cx. quinquefasciatus in Metropolitan Manila, Philippines.

METHODS: Adult Cx. quinquefasciatus mosquitoes were collected using a commercially available light-trap from May 2014-January 2015. Based on their sampling grids (n = 51), the adult mosquito abdomens were pooled and subjected to Wolbachia surface protein (wsp) gene amplification assay. Five selected wsp-positive samples were then sequenced and further analyzed to infer their phylogenetic relationship with known Wolbachia strains.

RESULTS: A total of 1090 adult Cx. quinquefasciatus mosquitoes were collected. Pooled abdomens (n = 53) were then sorted based on their sampling grids for subsequent screening of wsp gene. Wolbachia infection rate was 59% (31/53). These infections were located at 29 (57%) sampling grids, and were observed to be widely distributed in the study area. Phylogenetic analysis indicated that the sample sequences were Wolbachia pipientis isolated from known hosts, Cx. pipiens and Cx. quinquefasciatus belonging to supergroup B clade.

The study was able to demonstrate the prevalence and distribution of Wolbachia in Cx. quinquefasciatus in Metropolitan Manila, Philippines. The findings of this study are geared towards proposing a vector control program that utilizes the potential of Wolbachia as a biological control agent in preventing the transmission of Culex-borne diseases.}, } @article {pmid30992179, year = {2019}, author = {Muñoz-Leal, S and Macedo, C and Gonçalves, TC and Dias Barreira, J and Labruna, MB and de Lemos, ERS and Ogrzewalska, M}, title = {Detected microorganisms and new geographic records of Ornithodoros rietcorreai (Acari: Argasidae) from northern Brazil.}, journal = {Ticks and tick-borne diseases}, volume = {10}, number = {4}, pages = {853-861}, doi = {10.1016/j.ttbdis.2019.04.004}, pmid = {30992179}, issn = {1877-9603}, mesh = {Anaplasmataceae/genetics/isolation & purification ; Animals ; Babesia/genetics/isolation & purification ; Bacteria/*genetics/isolation & purification ; Borrelia/genetics/isolation & purification ; Brazil ; DNA, Bacterial/genetics/isolation & purification ; DNA, Protozoan/genetics/isolation & purification ; Female ; Geography ; Larva/microbiology ; Male ; Nymph/microbiology ; Ornithodoros/*microbiology/*parasitology ; *Phylogeny ; Rickettsia/genetics/isolation & purification ; }, abstract = {Reliable data on distributional ranges of soft ticks (Argasidae) and assessments of putative tick-borne agents enhance the understanding on tick-associated microorganisms. A total of 96 ticks morphologicaly and molecularly identified as Ornithodoros rietcorreai were collected in Tocantins State, Brazil, using Noireau traps with living bait as CO2 source. Ninety-six ticks (54 nymphs, 32 males, 10 females) with different engorgement degrees were collected. Fourty-seven (48.9%) of them were individually screened by PCR for detecting bacteria of Anaplasmataceae family and genera Rickettsia, and Borrelia. The presence of protozoans of the genus Babesia was assessed as well. Fourty seven ticks were submitted to analysis. Nine ticks (19.1%) yielded sequences for gltA and htrA genes most identical with a series of endosymbiont rickettsiae and Rickettsia bellii, respectively. Upon two ticks (4.2%) we retrieved DNA of a potential new Wolbachia sp., and DNA of a putative novel Hepatozoon was characterized from three (6.4%) specimens. No DNA of Babesia or Borrelia was detected. Remarkably, amplicons of unidentified eukaryotic organisms, most closely related with apicomplexans but also with dinoflagellates (91% of identity after BLAST analyses), were recovered from two ticks (4.2%) using primers designed for Babesia 18S rRNA gene. Our records expand the distribution of O. rietcorreai into Brazilian Cerrado biome and introduce the occurrence of microorganisms in this tick species.}, } @article {pmid30991952, year = {2019}, author = {Dhaygude, K and Nair, A and Johansson, H and Wurm, Y and Sundström, L}, title = {The first draft genomes of the ant Formica exsecta, and its Wolbachia endosymbiont reveal extensive gene transfer from endosymbiont to host.}, journal = {BMC genomics}, volume = {20}, number = {1}, pages = {301}, pmid = {30991952}, issn = {1471-2164}, support = {252411 and 284666 (Centre of Excellence in Biological Interactions)//Academy of Finland/ ; BB/K004204/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; NE/L00626X/1//Natural Environment Research Council/ ; }, mesh = {Animals ; Ants/*genetics/*microbiology ; Evolution, Molecular ; *Gene Transfer, Horizontal ; Genes, Insect/genetics ; *Genomics ; Symbiosis/*genetics ; Wolbachia/*genetics/*physiology ; }, abstract = {BACKGROUND: Adapting to changes in the environment is the foundation of species survival, and is usually thought to be a gradual process. However, transposable elements (TEs), epigenetic modifications, and/or genetic material acquired from other organisms by means of horizontal gene transfer (HGTs), can also lead to novel adaptive traits. Social insects form dense societies, which attract and maintain extra- and intracellular accessory inhabitants, which may facilitate gene transfer between species. The wood ant Formica exsecta (Formicidae; Hymenoptera), is a common ant species throughout the Palearctic region. The species is a well-established model for studies of ecological characteristics and evolutionary conflict.

RESULTS: In this study, we sequenced and assembled draft genomes for F. exsecta and its endosymbiont Wolbachia. The F. exsecta draft genome is 277.7 Mb long; we identify 13,767 protein coding genes, for which we provide gene ontology and protein domain annotations. This is also the first report of a Wolbachia genome from ants, and provides insights into the phylogenetic position of this endosymbiont. We also identified multiple horizontal gene transfer events (HGTs) from Wolbachia to F. exsecta. Some of these HGTs have also occurred in parallel in multiple other insect genomes, highlighting the extent of HGTs in eukaryotes.

CONCLUSION: We present the first draft genome of ant F. exsecta, and its endosymbiont Wolbachia (wFex), and show considerable rates of gene transfer from the symbiont to the host. We expect that especially the F. exsecta genome will be valuable resource in further exploration of the molecular basis of the evolution of social organization.}, } @article {pmid30991073, year = {2019}, author = {Zheng, B and Liu, X and Tang, M and Xi, Z and Yu, J}, title = {Use of age-stage structural models to seek optimal Wolbachia-infected male mosquito releases for mosquito-borne disease control.}, journal = {Journal of theoretical biology}, volume = {472}, number = {}, pages = {95-109}, doi = {10.1016/j.jtbi.2019.04.010}, pmid = {30991073}, issn = {1095-8541}, mesh = {Aedes/*growth & development/*microbiology ; Animals ; Female ; *Life Cycle Stages ; Male ; *Models, Biological ; *Mosquito Control ; Mosquito Vectors/*microbiology ; Reproducibility of Results ; Wolbachia/*physiology ; }, abstract = {Due to the lack of vaccines and effective clinical cures, current methods to control mosquito-borne viral diseases such as dengue and Zika are primarily targeting to eradicate the major mosquito vectors. However, traditional means, including larval source reduction and applications of insecticides etc, are not sufficient to keep vector population density below the epidemic risk threshold. An innovative and operational strategy is to release Wolbachia-infected male mosquitoes into wild areas to sterilize wild female mosquitoes by cytoplasmic incompatibility. To help design optimal release strategies before large scale and expensive operations, we started with an age-stage discrete model to track daily abundances of wild female mosquitoes, which fitted the field data collected by Guangzhou Center for Disease Control and Prevention from 2015 to 2017 with an average Pearson correlation coefficient 0.7283. Then, we modeled the Wolbachia interference by introducing the proportional releases of Wolbachia-infected males, and eight optimal release policies which guarantee more than 95% suppression efficiency were sought. Finally, we assessed the robustness of the optimality of the eight release policies by allowing the migration of females or the contamination of Wolbachia-infected females by two further extended mathematical models.}, } @article {pmid30976027, year = {2019}, author = {Lefoulon, E and Vaisman, N and Frydman, HM and Sun, L and Voland, L and Foster, JM and Slatko, BE}, title = {Large Enriched Fragment Targeted Sequencing (LEFT-SEQ) Applied to Capture of Wolbachia Genomes.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {5939}, pmid = {30976027}, issn = {2045-2322}, mesh = {Aedes/microbiology ; Animals ; Drosophila melanogaster/microbiology ; Evolution, Molecular ; *Gene Expression Regulation, Bacterial ; *Genome, Bacterial ; Genomics ; High-Throughput Nucleotide Sequencing/*methods ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Symbiosis is a major force of evolutionary change, influencing virtually all aspects of biology, from population ecology and evolution to genomics and molecular/biochemical mechanisms of development and reproduction. A remarkable example is Wolbachia endobacteria, present in some parasitic nematodes and many arthropod species. Acquisition of genomic data from diverse Wolbachia clades will aid in the elucidation of the different symbiotic mechanisms(s). However, challenges of de novo assembly of Wolbachia genomes include the presence in the sample of host DNA: nematode/vertebrate or insect. We designed biotinylated probes to capture large fragments of Wolbachia DNA for sequencing using PacBio technology (LEFT-SEQ: Large Enriched Fragment Targeted Sequencing). LEFT-SEQ was used to capture and sequence four Wolbachia genomes: the filarial nematode Brugia malayi, wBm, (21-fold enrichment), Drosophila mauritiana flies (2 isolates), wMau (11-fold enrichment), and Aedes albopictus mosquitoes, wAlbB (200-fold enrichment). LEFT-SEQ resulted in complete genomes for wBm and for wMau. For wBm, 18 single-nucleotide polymorphisms (SNPs), relative to the wBm reference, were identified and confirmed by PCR. A limit of LEFT-SEQ is illustrated by the wAlbB genome, characterized by a very high level of insertion sequences elements (ISs) and DNA repeats, for which only a 20-contig draft assembly was achieved.}, } @article {pmid30975197, year = {2019}, author = {Lee, WS and Webster, JA and Madzokere, ET and Stephenson, EB and Herrero, LJ}, title = {Mosquito antiviral defense mechanisms: a delicate balance between innate immunity and persistent viral infection.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {165}, pmid = {30975197}, issn = {1756-3305}, support = {DE140101493//Australian Research Council/ ; }, mesh = {Animals ; Culicidae/*immunology/*virology ; Humans ; Immunity, Innate ; Mosquito Control ; RNA Interference ; RNA, Viral ; }, abstract = {Mosquito-borne diseases are associated with major global health burdens. Aedes spp. and Culex spp. are primarily responsible for the transmission of the most medically important mosquito-borne viruses, including dengue virus, West Nile virus and Zika virus. Despite the burden of these pathogens on human populations, the interactions between viruses and their mosquito hosts remain enigmatic. Viruses enter the midgut of a mosquito following the mosquito's ingestion of a viremic blood meal. During infection, virus recognition by the mosquito host triggers their antiviral defense mechanism. Of these host defenses, activation of the RNAi pathway is the main antiviral mechanism, leading to the degradation of viral RNA, thereby inhibiting viral replication and promoting viral clearance. However, whilst antiviral host defense mechanisms limit viral replication, the mosquito immune system is unable to effectively clear the virus. As such, these viruses can establish persistent infection with little or no fitness cost to the mosquito vector, ensuring life-long transmission to humans. Understanding of the mosquito innate immune response enables the discovery of novel antivectorial strategies to block human transmission. This review provides an updated and concise summary of recent studies on mosquito antiviral immune responses, which is a key determinant for successful virus transmission. In addition, we will also discuss the factors that may contribute to persistent infection in mosquito hosts. Finally, we will discuss current mosquito transmission-blocking strategies that utilize genetically modified mosquitoes and Wolbachia-infected mosquitoes for resistance to pathogens.}, } @article {pmid30970277, year = {2019}, author = {Sigle, LT and McGraw, EA}, title = {Expanding the canon: Non-classical mosquito genes at the interface of arboviral infection.}, journal = {Insect biochemistry and molecular biology}, volume = {109}, number = {}, pages = {72-80}, doi = {10.1016/j.ibmb.2019.04.004}, pmid = {30970277}, issn = {1879-0240}, mesh = {Animals ; Arboviruses/*physiology ; Culicidae/genetics/*immunology/virology ; Immunity, Innate/*genetics ; Insect Proteins/*genetics/metabolism ; Mice ; Mosquito Vectors/genetics/*immunology/virology ; *Transcriptome ; }, abstract = {Mosquito transmitted viruses cause significant morbidity and mortality in human populations. Despite the use of insecticides and other measures of vector control, arboviral diseases are on the rise. One potential solution for limiting disease transmission to humans is to render mosquitoes refractory to viral infection through genetic modification. Substantial research effort in Drosophila, Aedes and Anopheles has helped to define the major innate immune pathways, including Toll, IMD, Jak/Stat and RNAi, however we still have an incomplete picture of the mosquito antiviral response. Transcriptional profiles of virus-infected insects reveal a much wider range of pathways activated by the process of infection. Within these lists of genes are unexplored mosquito candidates of viral defense. Wolbachia species are endosymbiotic bacteria that naturally limit arboviral infection in mosquitoes. Our understanding of the Wolbachia-mediated viral blocking mechanism is poor, but it does not appear to operate via the classical immune pathways. Herein, we reviewed the transcriptomic response of mosquitoes to multiple viral species and put forth consensus gene types/families outside the immune canon whose expression responds to infection, including cytoskeleton and cellular trafficking, the heat shock response, cytochromes P450, cell proliferation, chitin and small RNAs. We then examine emerging evidence for their functional role in viral resistance in diverse insect and mammalian hosts and their potential role in Wolbachia-mediated viral blocking. These candidate gene families offer novel avenues for research into the nature of insect viral defense.}, } @article {pmid30958712, year = {2019}, author = {Clare, RH and Clark, R and Bardelle, C and Harper, P and Collier, M and Johnston, KL and Plant, H and Plant, D and McCall, E and Slatko, BE and Cantin, L and Wu, B and Ford, L and Murray, D and Rich, K and Wigglesworth, M and Taylor, MJ and Ward, SA}, title = {Development of a High-Throughput Cytometric Screen to Identify Anti- Wolbachia Compounds: The Power of Public-Private Partnership.}, journal = {SLAS discovery : advancing life sciences R & D}, volume = {24}, number = {5}, pages = {537-547}, pmid = {30958712}, issn = {2472-5560}, support = {MC_PC_17167/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Anti-Bacterial Agents/chemistry/*pharmacology ; Cell Culture Techniques/methods ; Drug Discovery ; Elephantiasis, Filarial/drug therapy ; *High-Throughput Screening Assays ; Host-Pathogen Interactions/*drug effects ; Humans ; Image Cytometry ; Onchocerciasis/drug therapy ; Wolbachia/*drug effects/pathogenicity/ultrastructure ; }, abstract = {The Anti- Wolbachia (A·WOL) consortium at the Liverpool School of Tropical Medicine (LSTM) has partnered with the Global High-Throughput Screening (HTS) Centre at AstraZeneca to create the first anthelmintic HTS for neglected tropical diseases (NTDs). The A·WOL consortium aims to identify novel macrofilaricidal drugs targeting the essential bacterial symbiont (Wolbachia) of the filarial nematodes causing onchocerciasis and lymphatic filariasis. Working in collaboration, we have validated a robust high-throughput assay capable of identifying compounds that selectively kill Wolbachia over the host insect cell. We describe the development and validation process of this complex, phenotypic high-throughput assay and provide an overview of the primary outputs from screening the AstraZeneca library of 1.3 million compounds.}, } @article {pmid30953430, year = {2019}, author = {Landmann, F}, title = {The Wolbachia Endosymbionts.}, journal = {Microbiology spectrum}, volume = {7}, number = {2}, pages = {}, doi = {10.1128/microbiolspec.BAI-0018-2019}, pmid = {30953430}, issn = {2165-0497}, mesh = {Animals ; Arthropods/microbiology ; Filarioidea/microbiology ; Host Microbial Interactions/*physiology ; Insecta/microbiology ; Symbiosis ; Wolbachia/classification/genetics/pathogenicity/*physiology ; }, abstract = {The Wolbachia endosymbionts encompass a large group of intracellular bacteria of biomedical and veterinary relevance, closely related to Anaplasma, Ehrlichia, and Rickettsia. This genus of Gram-negative members of the Alphaproteobacteria does not infect vertebrates but is instead restricted to ecdysozoan species, including terrestrial arthropods and a family of parasitic filarial nematodes, the Onchocercidae. The Wolbachia profoundly impact not only the ecology and evolution but also the reproductive biology of their hosts, through a wide range of symbiotic interactions. Because they are essential to the survival and reproduction of their filarial nematode hosts, they represent an attractive target to fight filariasis. Their abilities to spread through insect populations and to affect vector competence through pathogen protection have made Wolbachia a staple for controlling vector-borne diseases. Estimated to be present in up to 66% of insect species, the Wolbachia are probably the most abundant endosymbionts on earth. Their success resides in their unique capacity to infect and manipulate the host germ line to favor their vertical transmission through the maternal lineage. Because the Wolbachia resist genetic manipulation and growth in axenic culture, our understanding of their biology is still in its infancy. Despite these limitations, the "-omics" revolution combined with the use of well-established and emerging experimental host models is accelerating our comprehension of the host phenotypes caused by Wolbachia, and the identification of Wolbachia effectors is ongoing.}, } @article {pmid30950399, year = {2019}, author = {Walden, PM and Whitten, AE and Premkumar, L and Halili, MA and Heras, B and King, GJ and Martin, JL}, title = {The atypical thiol-disulfide exchange protein α-DsbA2 from Wolbachia pipientis is a homotrimeric disulfide isomerase.}, journal = {Acta crystallographica. Section D, Structural biology}, volume = {75}, number = {Pt 3}, pages = {283-295}, pmid = {30950399}, issn = {2059-7983}, support = {FL0992138//Australian Research Council/ ; }, mesh = {Bacterial Proteins/*chemistry ; Disulfides/*chemistry ; Escherichia coli K12/enzymology ; Protein Disulfide-Isomerases/*chemistry ; Scattering, Small Angle ; Wolbachia/*enzymology ; }, abstract = {Disulfide-bond-forming (DSB) oxidative folding enzymes are master regulators of virulence that are localized to the periplasm of many Gram-negative bacteria. The archetypal DSB machinery from Escherichia coli K-12 consists of a dithiol-oxidizing redox-relay pair (DsbA/B), a disulfide-isomerizing redox-relay pair (DsbC/D) and the specialist reducing enzymes DsbE and DsbG that also interact with DsbD. By contrast, the Gram-negative bacterium Wolbachia pipientis encodes just three DSB enzymes. Two of these, α-DsbA1 and α-DsbB, form a redox-relay pair analogous to DsbA/B from E. coli. The third enzyme, α-DsbA2, incorporates a DsbA-like sequence but does not interact with α-DsbB. In comparison to other DsbA enzymes, α-DsbA2 has ∼50 extra N-terminal residues (excluding the signal peptide). The crystal structure of α-DsbA2ΔN, an N-terminally truncated form in which these ∼50 residues are removed, confirms the DsbA-like nature of this domain. However, α-DsbA2 does not have DsbA-like activity: it is structurally and functionally different as a consequence of its N-terminal residues. Firstly, α-DsbA2 is a powerful disulfide isomerase and a poor dithiol oxidase: i.e. its role is to shuffle rather than to introduce disulfide bonds. Moreover, small-angle X-ray scattering (SAXS) of α-DsbA2 reveals a homotrimeric arrangement that differs from those of the other characterized bacterial disulfide isomerases DsbC from Escherichia coli (homodimeric) and ScsC from Proteus mirabilis (PmScsC; homotrimeric with a shape-shifter peptide). α-DsbA2 lacks the shape-shifter motif and SAXS data suggest that it is less flexible than PmScsC. These results allow conclusions to be drawn about the factors that are required for functionally equivalent disulfide isomerase enzymatic activity across structurally diverse protein architectures.}, } @article {pmid30949677, year = {2019}, author = {Pillonel, T and Bertelli, C and Aeby, S and de Barsy, M and Jacquier, N and Kebbi-Beghdadi, C and Mueller, L and Vouga, M and Greub, G}, title = {Sequencing the Obligate Intracellular Rhabdochlamydia helvetica within Its Tick Host Ixodes ricinus to Investigate Their Symbiotic Relationship.}, journal = {Genome biology and evolution}, volume = {11}, number = {4}, pages = {1334-1344}, pmid = {30949677}, issn = {1759-6653}, mesh = {Animals ; Chlamydiales/*genetics/metabolism ; Female ; Gene Transfer, Horizontal ; *Genome, Bacterial ; *Host-Parasite Interactions ; Ixodes/*microbiology ; Symbiosis ; }, abstract = {The Rhabdochlamydiaceae family is one of the most widely distributed within the phylum Chlamydiae, but most of its members remain uncultivable. Rhabdochlamydia 16S rRNA was recently reported in more than 2% of 8,534 pools of ticks from Switzerland. Shotgun metagenomics was performed on a pool of five female Ixodes ricinus ticks presenting a high concentration of chlamydial DNA, allowing the assembly of a high-quality draft genome. About 60% of sequence reads originated from a single bacterial population that was named "Candidatus Rhabdochlamydia helvetica" whereas only few thousand reads mapped to the genome of "Candidatus Midichloria mitochondrii," a symbiont normally observed in all I. ricinus females. The 1.8 Mbp genome of R. helvetica is smaller than other Chlamydia-related bacteria. Comparative analyses with other chlamydial genomes identified transposases of the PD-(D/E)XK nuclease family that are unique to this new genome. These transposases show evidence of interphylum horizontal gene transfers between multiple arthropod endosymbionts, including Cardinium spp. (Bacteroidetes) and diverse proteobacteria such as Wolbachia, Rickettsia spp. (Rickettsiales), and Caedimonas varicaedens (Holosporales). Bacterial symbionts were previously suggested to provide B-vitamins to hematophagous hosts. However, incomplete metabolic capacities including for B-vitamin biosynthesis, high bacterial density and limited prevalence suggest that R. helvetica is parasitic rather than symbiotic to its host. The identification of novel Rhabdochlamydia strains in different hosts and their sequencing will help understanding if members of this genus have become highly specialized parasites with reduced genomes, like the Chlamydiaceae, or if they could be pathogenic to humans using ticks as a transmission vector.}, } @article {pmid30941647, year = {2019}, author = {Barradas, I and Vázquez, V}, title = {Backward Bifurcation as a Desirable Phenomenon: Increased Fecundity Through Infection.}, journal = {Bulletin of mathematical biology}, volume = {81}, number = {6}, pages = {2029-2050}, doi = {10.1007/s11538-019-00604-1}, pmid = {30941647}, issn = {1522-9602}, mesh = {Animals ; Basic Reproduction Number/statistics & numerical data ; Biological Control Agents ; Communicable Disease Control/*methods/statistics & numerical data ; Epidemics/prevention & control/statistics & numerical data ; Epidemiologic Methods ; Female ; *Fertility ; Infections/*physiopathology ; Insect Vectors/microbiology ; Male ; Mathematical Concepts ; *Models, Biological ; Population Density ; Symbiosis ; Wolbachia/pathogenicity/physiology ; }, abstract = {Backward or subcritical bifurcation is usually considered an undesirable phenomenon in epidemiology since control measures require a reduction in R0 not below one but below a much smaller value. However, there are contexts for which a backward or subcritical bifurcation is not a bad thing; it can even be desirable. Such is the case for any characteristic that can be passed to the next generation (genetically fixed or not) and that increases the effective reproductive rate of the host or the total number of individuals. In the present work, we study an epidemiological model consisting of two classes, susceptible and "infected" individuals; the model considers a characteristic that is passed from "infected" to "susceptible" by direct "contact," for instance increased fecundity. We analyze conditions for the appearance of a backward or subcritical bifurcation. We discuss the advantage for the population under infection, since the total number of individuals increases at equilibrium. If one takes that as a proxy for increased fitness, it would increase the species' ecological success. One key element in the model is the fact that "susceptible" individuals have "susceptible" descendants, but "infected" individuals can have "infected" descendants as well as "susceptible" ones. A somehow rare addition for epidemiological models, the fact that "infected" individuals reproduce more rapidly than the susceptible ones, leads to unexpected consequences. Facilitating the "inoculation" increases the total population size, i.e., the backward or subcritical bifurcation appears, with desirable consequences for the population. We show that an increase in the number of susceptible newborns is the main reason for the appearance of a backward or subcritical bifurcation, which induces a bigger population size. We analyze the effect of different combinations of susceptible/infected birth rates. This kind of phenomenon has been observed for bacterial infections in several insects-bacteria and nematodes-bacteria interactions; in particular, it has been intensely studied in interactions of wasps and flies with the genus Wolbachia. It has also been shown in amphibians.}, } @article {pmid30933079, year = {2019}, author = {Yang, S and Zhao, Y and Yu, J and Fan, Z and Gong, ST and Tang, H and Pan, L}, title = {Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {145}, pages = {}, doi = {10.3791/58845}, pmid = {30933079}, issn = {1940-087X}, mesh = {Animals ; Antiviral Agents/metabolism ; Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster/*virology ; *Host-Pathogen Interactions ; Immunity, Innate/immunology ; Mutation/genetics ; RNA Interference ; Reproducibility of Results ; Signal Transduction ; Viral Load ; Virus Diseases/immunology/*virology ; Viruses/metabolism ; Wolbachia/physiology ; }, abstract = {Virus spreading is a major cause of epidemic diseases. Thus, understanding the interaction between the virus and the host is very important to extend our knowledge of prevention and treatment of viral infection. The fruit fly Drosophila melanogaster has proven to be one of the most efficient and productive model organisms to screen for antiviral factors and investigate virus-host interaction, due to powerful genetic tools and highly conserved innate immune signaling pathways. The procedure described here demonstrates a nano-injection method to establish viral infection and induce systemic antiviral responses in adult flies. The precise control of the viral injection dose in this method enables high experimental reproducibility. Protocols described in this study include the preparation of flies and the virus, the injection method, survival rate analysis, the virus load measurement, and an antiviral pathway assessment. The influence effects of viral infection by the flies' background were mentioned here. This infection method is easy to perform and quantitatively repeatable; it can be applied to screen for host/viral factors involved in virus-host interaction and to dissect the crosstalk between innate immune signaling and other biological pathways in response to viral infection.}, } @article {pmid30930872, year = {2019}, author = {Ali, H and Muhammad, A and Sanda, NB and Huang, Y and Hou, Y}, title = {Pyrosequencing Uncovers a Shift in Bacterial Communities Across Life Stages of Octodonta nipae (Coleoptera: Chrysomelidae).}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {466}, pmid = {30930872}, issn = {1664-302X}, abstract = {Bacterial symbionts of insects affect a wide array of host traits including fitness and immunity. Octodonta nipae (Maulik), commonly known as hispid leaf beetle is a destructive palm pest around the world. Understanding the dynamics of microbiota is essential to unravel the complex interplay between O. nipae and its bacterial symbionts. In this study, bacterial 16S rRNA V3-V4 region was targeted to decipher the diversity and dynamics of bacterial symbionts across different life stages [eggs, larvae, pupae, and adult (male and female)] and reproductive organs (ovaries and testis) of O. nipae. Clustering analysis at ≥97% similarity threshold produced 3,959 operational taxonomic units (OTUs) that belonged to nine different phyla. Proteobacteria, Actinobacteria, and Firmicutes represented the bulk of taxa that underwent notable changes during metamorphosis. Enterobacteriaceae and Dermabacteraceae were the most abundant families in immature stages (eggs, larvae, and pupae), while Anaplasmataceae family was dominated in adults (male and female) and reproductive organs (ovaries and testis). The genus Serratia and Lactococcus were most abundant in eggs, whereas Pantoea and Brachybacterium represented the bulk of larvae and pupae microbiota. Interestingly the genus Wolbachia found positive to all tested samples and was recorded extremely high (>64%) in the adults and reproductive organs. The bacteria varied across the developmental stages and responsible for various metabolic activities. Selection choice exerted by the insect host as a result of its age or developmental stage could be the main reason to ascertain the shift in the bacteria populations. Maternally inherited Wolbachia was found to be an obligate endosymbiont infecting all tested life stages, body parts, and tissues. These outcomes foster our understanding of the intricate associations between bacteria and O. nipae and will incorporate in devising novel pest control strategies against this palm pest.}, } @article {pmid30924288, year = {2020}, author = {Zhang, X and Li, TP and Zhou, CY and Zhao, DS and Zhu, YX and Bing, XL and Huang, HJ and Hong, XY}, title = {Antibiotic exposure perturbs the bacterial community in the small brown planthopper Laodelphax striatellus.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {895-907}, doi = {10.1111/1744-7917.12675}, pmid = {30924288}, issn = {1744-7917}, support = {804015//Nanjing Agricultural University/ ; 31672035//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Bacteria/*drug effects ; Female ; Hemiptera/*microbiology ; High-Throughput Nucleotide Sequencing ; Microbiota/*drug effects ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; Tetracycline/*pharmacology ; Tissue Distribution ; }, abstract = {Bacteria symbionts in herbivores play an important role in host biology and ecology, and are affected by environmental factors such as temperature, diet, habitat, antibiotics and so on. However, the effects of antibiotics on the microbiome of the small brown planthopper Laodelphax striatellus (SBPH) remain unclear. Here, we studied the effects of tetracycline on the diversity and composition of bacterial colonies in different tissues of SBPH using high throughput sequencing of 16S ribosomal RNA amplicons. Our results show that Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria were most abundant in SBPH, and the genera Asaia and Wolbachia were most abundant in all body parts of SBPH. Antibiotic treatment had persistent effects on the composition of the SBPH microbiome. Tetracycline depleted the population of Firmicutes, Bacteroidetes, Tenericutes and Fusobacteria, and nearly 100% eliminated Wolbachia, Bacteroides and Abiotrophia in SBPH. Together, these results suggest that antibiotic exposure affects the bacteria symbionts of different body parts in SBPH and will facilitate future studies of the bacterial symbionts of arthropod hosts.}, } @article {pmid30915518, year = {2019}, author = {Kolasa, M and Ścibior, R and Mazur, MA and Kubisz, D and Dudek, K and Kajtoch, Ł}, title = {How Hosts Taxonomy, Trophy, and Endosymbionts Shape Microbiome Diversity in Beetles.}, journal = {Microbial ecology}, volume = {78}, number = {4}, pages = {995-1013}, pmid = {30915518}, issn = {1432-184X}, support = {DEC-2013/11/D/NZ8/00583//National Science Centre, Poland/ ; small grants for young researchers//Polish Ministry of Science and Higher Education/ ; }, mesh = {Animals ; Bacteria/*classification ; Bacterial Physiological Phenomena ; Coleoptera/classification/*microbiology/*physiology ; Feeding Behavior ; Microbiota/*physiology ; Phylogeny ; *Symbiosis ; }, abstract = {Bacterial communities play a crucial role in the biology, ecology, and evolution of multicellular organisms. In this research, the microbiome of 24 selected beetle species representing five families (Carabidae, Staphylinidae, Curculionidae, Chrysomelidae, Scarabaeidae) and three trophic guilds (carnivorous, herbivorous, detrivorous) was examined using 16S rDNA sequencing on the Illumina platform. The aim of the study was to compare diversity within and among species on various levels of organization, including evaluation of the impact of endosymbiotic bacteria. Collected data showed that beetles possess various bacterial communities and that microbiota of individuals of particular species hosts are intermixed. The most diverse microbiota were found in Carabidae and Scarabaeidae; the least diverse, in Staphylinidae. On higher organization levels, the diversity of bacteria was more dissimilar between families, while the most distinct with respect to their microbiomes were trophic guilds. Moreover, eight taxa of endosymbiotic bacteria were detected including common genera such as Wolbachia, Rickettsia, and Spiroplasma, as well as the rarely detected Cardinium, Arsenophonus, Buchnera, Sulcia, Regiella, and Serratia. There were no correlations among the abundance of the most common Wolbachia and Rickettsia; a finding that does not support the hypothesis that these bacteria occur interchangeably. The abundance of endosymbionts only weakly and negatively correlates with diversity of the whole microbiome in beetles. Overall, microbiome diversity was found to be more dependent on host phylogeny than on the abundance of endosymbionts. This is the first study in which bacteria diversity is compared between numerous species of beetles in a standardized manner.}, } @article {pmid30914397, year = {2019}, author = {Konecka, E and Olszanowski, Z and Koczura, R}, title = {Wolbachia of phylogenetic supergroup E identified in oribatid mite Gustavia microcephala (Acari: Oribatida).}, journal = {Molecular phylogenetics and evolution}, volume = {135}, number = {}, pages = {230-235}, doi = {10.1016/j.ympev.2019.03.019}, pmid = {30914397}, issn = {1095-9513}, mesh = {Animals ; Base Sequence ; DNA, Ribosomal/genetics ; Likelihood Functions ; Mites/*microbiology ; *Phylogeny ; Wolbachia/*classification/genetics ; }, abstract = {Heritable endosymbionts have been observed in arthropod and nematode hosts. The most-known among them is Wolbachia. Although the bacterium was previously identified in oribatid mites (Acari: Oribatida), it was not assigned to any phylogenetic group. Endosymbionts have a profound influence on their hosts, playing various functions that affect invertebrate's biology such as changing the way of reproduction. Oribatida provide the very unique examples of groups in which even whole families appear to be thelytokous, so we considered that it is worth to investigate the occurrence of endosymbiotic microorganisms in oribatid mites, especially that the knowledge on the symbionts occurrence in this invertebrate group is negligible. We report for the first time Wolbachia in oribatid mite Gustavia microcephala. The sequences of 16S rDNA, gltA, and ftsZ genes of the endosymbiont from the mite showed the highest similarity to Wolbachia found in Collembola. Phylogenetic analysis based on single gene and concatenated alignments of three genes revealed that the bacteria from G. microcephala and Collembola were related and clustered together with supergroup E. Relatively close relationship of Wolbachia from oribatid and collembolan hosts might mean at the evolutionary scale that horizontal transfer of bacteria between these two groups of invertebrates may take place.}, } @article {pmid30912316, year = {2019}, author = {Guo, J and Liu, X and Poncelet, N and He, K and Francis, F and Wang, Z}, title = {Detection and geographic distribution of seven facultative endosymbionts in two Rhopalosiphum aphid species.}, journal = {MicrobiologyOpen}, volume = {8}, number = {8}, pages = {e00817}, pmid = {30912316}, issn = {2045-8827}, mesh = {Animals ; Aphids/*microbiology ; China ; Europe ; Gammaproteobacteria/classification/*isolation & purification ; *Phylogeography ; Rickettsiales/classification/*isolation & purification ; Spiroplasma/classification/*isolation & purification ; *Symbiosis ; }, abstract = {Study of the mutualistic associations between facultative symbionts and aphids are developed only in a few models. That survey on the situation and distribution of the symbionts in a certain area is helpful to obtain clues for the acquisition and spread of them as well as their roles played in host evolution. To understand the infection patterns of seven facultative symbionts (Serratia symbiotica, Hamiltonella defensa, Regiella insecticola, Rickettsia, Spiroplasma, Wolbachia, and Arsenophonus) in Rhopalosiphum padi (Linnaeus) and Rhopalosiphum maidis (Fitch), we collected 882 R. maidis samples (37 geographical populations) from China and 585 R. padi samples (32 geographical populations) from China and Europe. Results showed that both species were widely infected with various symbionts and totally 50.8% of R. maidis and 50.1% of R. padi were multi-infected with targeted symbionts. However, very few Rhopalosiphum aphids were infected with S. symbiotica. The infection frequencies of some symbionts were related to the latitude of collecting sites, suggesting the importance of environmental factors in shaping the geographic distribution of facultative symbionts. Also, R. maidis and R. padi were infected with different H. defensa strains based on phylogenetic analysis which may be determined by host ×symbiont genotype interactions. According to our results, the ubiquitous symbionts may play important roles in the evolution of their host aphid and their impacts on adaptation of R. padi and R. maidis were discussed as well.}, } @article {pmid30912174, year = {2019}, author = {Bing, XL and Zhao, DS and Hong, XY}, title = {Bacterial reproductive manipulators in rice planthoppers.}, journal = {Archives of insect biochemistry and physiology}, volume = {101}, number = {2}, pages = {e21548}, doi = {10.1002/arch.21548}, pmid = {30912174}, issn = {1520-6327}, support = {31672035//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; 804015//the startup grant from Nanjing Agricultural University/ ; }, mesh = {Animals ; *Bacterial Physiological Phenomena ; Female ; Hemiptera/*microbiology/physiology ; Male ; Reproduction/*physiology ; Symbiosis ; }, abstract = {Rice planthoppers (Hemiptera: Delphacidae) are notorious pests for rice (Oryza sativa) in Asia, posing a serious threat to rice production and grain security. Rice planthoppers harbor diverse bacterial symbionts, including Wolbachia, Cardinium, Spiroplasma, and Arsenophonus, which are known to manipulate reproduction in arthropod hosts. This microreview is to introduce current knowledge of bacterial reproductive manipulators in rice planthoppers, including their diversity, population dynamics, localization, transmission, and biological functions.}, } @article {pmid30894837, year = {2019}, author = {Tolley, SJA and Nonacs, P and Sapountzis, P}, title = {Wolbachia Horizontal Transmission Events in Ants: What Do We Know and What Can We Learn?.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {296}, pmid = {30894837}, issn = {1664-302X}, abstract = {While strict vertical transmission insures the durability of intracellular symbioses, phylogenetic incongruences between hosts and endosymbionts suggest horizontal transmission must also occur. These horizontal acquisitions can have important implications for the biology of the host. Wolbachia is one of the most ecologically successful prokaryotes in arthropods, infecting an estimated 50-70% of all insect species. Much of this success is likely due to the fact that, in arthropods, Wolbachia is notorious for manipulating host reproduction to favor transmission through the female germline. However, its natural potential for horizontal transmission remains poorly understood. Here we evaluate the fundamental prerequisites for successful horizontal transfer, including necessary environmental conditions, genetic potential of bacterial strains, and means of mediating transfers. Furthermore, we revisit the relatedness of Wolbachia strains infecting the Panamanian leaf-cutting ant, Acromyrmex echinatior, and its inquiline social parasite, Acromyrmex insinuator, and compare our results to a study published more than 15 years ago by Van Borm et al. (2003). The results of this pilot study prompt us to reevaluate previous notions that obligate social parasitism reliably facilitates horizontal transfer and suggest that not all Wolbachia strains associated with ants have the same genetic potential for horizontal transmission.}, } @article {pmid30893296, year = {2019}, author = {Fattouh, N and Cazevieille, C and Landmann, F}, title = {Wolbachia endosymbionts subvert the endoplasmic reticulum to acquire host membranes without triggering ER stress.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {3}, pages = {e0007218}, pmid = {30893296}, issn = {1935-2735}, mesh = {Animals ; Cell Line ; Drosophila melanogaster/cytology/*microbiology ; Endoplasmic Reticulum/metabolism/*microbiology ; Gene Expression Profiling ; Golgi Apparatus/metabolism/microbiology ; Host-Pathogen Interactions ; Intracellular Membranes/metabolism/*microbiology ; Stress, Physiological/genetics/*physiology ; Symbiosis/genetics/*physiology ; Vacuoles/microbiology ; Wolbachia/pathogenicity/*physiology ; }, abstract = {The reproductive parasites Wolbachia are the most common endosymbionts on earth, present in a plethora of arthropod species. They have been introduced into mosquitos to successfully prevent the spread of vector-borne diseases, yet the strategies of host cell subversion underlying their obligate intracellular lifestyle remain to be explored in depth in order to gain insights into the mechanisms of pathogen-blocking. Like some other intracellular bacteria, Wolbachia reside in a host-derived vacuole in order to replicate and escape the immune surveillance. Using here the pathogen-blocking Wolbachia strain from Drosophila melanogaster, introduced into two different Drosophila cell lines, we show that Wolbachia subvert the endoplasmic reticulum to acquire their vacuolar membrane and colonize the host cell at high density. Wolbachia redistribute the endoplasmic reticulum, and time lapse experiments reveal tight coupled dynamics suggesting important signalling events or nutrient uptake. Wolbachia infection however does not affect the tubular or cisternal morphologies. A fraction of endoplasmic reticulum becomes clustered, allowing the endosymbionts to reside in between the endoplasmic reticulum and the Golgi apparatus, possibly modulating the traffic between these two organelles. Gene expression analyses and immunostaining studies suggest that Wolbachia achieve persistent infections at very high titers without triggering endoplasmic reticulum stress or enhanced ERAD-driven proteolysis, suggesting that amino acid salvage is achieved through modulation of other signalling pathways.}, } @article {pmid30889242, year = {2019}, author = {Endersby-Harshman, NM and Axford, JK and Hoffmann, AA}, title = {Environmental Concentrations of Antibiotics May Diminish Wolbachia infections in Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {56}, number = {4}, pages = {1078-1086}, doi = {10.1093/jme/tjz023}, pmid = {30889242}, issn = {1938-2928}, mesh = {Aedes/drug effects/*microbiology ; Animals ; Chlortetracycline/*toxicity ; Female ; Larva/drug effects ; Male ; Water Pollutants, Chemical/*toxicity ; Wolbachia/*drug effects ; }, abstract = {Wolbachia-infected Aedes aegypti (L.) mosquitoes for control of dengue transmission are being released experimentally in tropical regions of Australia, south-east Asia, and South America. To become established, the Wolbachia Hertig (Rickettsiales: Rickettsiaceae) strains used must induce expression of cytoplasmic incompatibility (CI) in matings between infected males and uninfected females so that infected females have a reproductive advantage, which will drive the infection through field populations. Wolbachia is a Rickettsia-like alphaproteobacterium which can be affected by tetracycline antibiotics. We investigated whether exposure of Wolbachia-infected mosquitoes to chlortetracycline at environmentally relevant levels during their aquatic development resulted in loss or reduction of infection in three strains, wAlbB, wMel, and wMelPop. Wolbachia density was reduced for all three strains at the tested chlortetracycline concentrations of 5 and 50 µg/liter. Two of the strains, wMel and wMelPop, showed a breakdown in CI. The wAlbB strain maintained CI and may be useful at breeding sites where tetracycline contamination has occurred. This may include drier regions where Ae. aegypti can utilize subterranean water sources and septic tanks as breeding sites.}, } @article {pmid30889229, year = {2019}, author = {Portillo, A and Palomar, AM and de Toro, M and Santibáñez, S and Santibáñez, P and Oteo, JA}, title = {Exploring the bacteriome in anthropophilic ticks: To investigate the vectors for diagnosis.}, journal = {PloS one}, volume = {14}, number = {3}, pages = {e0213384}, pmid = {30889229}, issn = {1932-6203}, mesh = {Animals ; Arachnid Vectors/*microbiology ; DNA, Bacterial/genetics ; Dermacentor/microbiology ; Humans ; Ixodes/microbiology ; Ixodidae/*microbiology ; *Metagenome ; Metagenomics ; Proteobacteria/classification/genetics ; RNA, Ribosomal, 16S/genetics ; Rhipicephalus sanguineus/microbiology ; Spain ; Tick Bites/microbiology ; Tick-Borne Diseases/diagnosis/microbiology ; }, abstract = {OBJECTIVE: The aim of this study was to characterize the bacterial microbiome of hard ticks with affinity to bite humans in La Rioja (North of Spain).

METHODS: A total of 88 adult ticks (22 Rhipicephalus sanguineus sensu lato, 27 Haemaphysalis punctata, 30 Dermacentor marginatus and 9 Ixodes ricinus) and 120 I. ricinus nymphs (CRETAV collection, La Rioja, Spain), representing the main anthropophilic species in our environment, were subjected to a metagenomic analysis of the V3-V4 region of the 16S rRNA gene using an Illumina MiSeq platform. Data obtained with Greengenes database were refined with BLAST. Four groups of samples were defined, according to the four tick species.

RESULTS: Proteobacteria was the predominant phylum observed in all groups. Gammaproteobacteria was the most abundant class, followed by Alphaproteobacteria for R. sanguineus, H. punctata and D. marginatus but the relative abundance of reads for these classes was reversed for I. ricinus. This tick species showed more than 46% reads corresponding to 'not assigned' OTUs (Greengenes), and >97% of them corresponded to 'Candidatus Midichloriaceae' using BLAST. Within Rickettsiales, 'Candidatus Midichloria', Rickettsia, Ehrlichia, 'Candidatus Neoehrlichia' and Wolbachia were detected. I. ricinus was the most alpha-diverse species. Regarding beta-diversity, I. ricinus and H. punctata samples grouped according to their tick species but microbial communities of some R. sanguineus and D. marginatus specimens clustered together.

CONCLUSIONS: The metagenomics approach seems useful to discover the spectrum of tick-related bacteria. More studies are needed to identify and differentiate bacterial species, and to improve the knowledge of tick-borne diseases in Spain.}, } @article {pmid30889213, year = {2019}, author = {Kanakala, S and Ghanim, M}, title = {Global genetic diversity and geographical distribution of Bemisia tabaci and its bacterial endosymbionts.}, journal = {PloS one}, volume = {14}, number = {3}, pages = {e0213946}, pmid = {30889213}, issn = {1932-6203}, mesh = {Animals ; Bacteria/genetics ; Bayes Theorem ; Genetic Variation ; Hemiptera/classification/*genetics/*microbiology ; Host Microbial Interactions/genetics ; Insect Control ; Phylogeny ; Symbiosis/genetics ; }, abstract = {Bemisia tabaci is one of the most threatening pests in agriculture, causing significant losses to many important crops on a global scale. The dramatic increase and availability of sequence data for B. tabaci species complex and its bacterial endosymbionts is critical for developing emerging sustainable pest management strategies which are based on pinpointing the global diversity of this important pest and its bacterial endosymbionts. To unravel the global genetic diversity of B. tabaci species complex focusing on its associated endosymbionts, along with Israeli whitefly populations collected in this study, we combined available sequences in databases, resulting in a total of 4,253 mitochondrial cytochrome oxidase I (mtCOI) sequences from 82 countries and 1,226 16S/23S rRNA endosymbiont sequences from 32 countries that were analyzed. Using Bayesian phylogenetic analysis, we identified two new B. tabaci groups within the species complex and described the global distribution of endosymbionts within this complex. Our analyses revealed complex divergence of the different endosymbiont sequences within the species complex, with overall one Hamiltonella, two Porteria (P1 and P2), two Arsenophonus (A1 and A2), two Wolbachia (super-groups O and B), four Cardinium (C1-C4) and three Rickettsia (R1-R3) groups were identified. Our comprehensive analysis provides an updated important resource for this globally important pest and its secondary symbionts, which have been a major subject for research in last three decades.}, } @article {pmid30869785, year = {2019}, author = {Balaji, S and Jayachandran, S and Prabagaran, SR}, title = {Evidence for the natural occurrence of Wolbachia in Aedes aegypti mosquitoes.}, journal = {FEMS microbiology letters}, volume = {366}, number = {6}, pages = {}, doi = {10.1093/femsle/fnz055}, pmid = {30869785}, issn = {1574-6968}, mesh = {Aedes/growth & development/*microbiology/physiology ; Animals ; Female ; India ; Male ; Phylogeny ; Reproduction ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia, a Gram-negative bacterium, is widely known to be present in arthropods and nematodes. Of late, great impetus is given to employ this intracellular bacterium, as an alternative to conventional biocontrol agents for the control of mosquitoes because of its inherent ability to induce sperm-egg incompatibility, feminisation etc. By employing molecular tools, we have shown the presence of Wolbachia from Aedes aegypti mosquito population collected from Coimbatore, India by PCR amplifying the Ae. aegypti mosquito genome with Wolbachia specific 16S rRNA, wsp and ftsZ gene primers. The phylogenetic analysis of these gene sequences incorporating MLST and GenBank reference sequences has confirmed the occurrence of Wolbachia supergroup B in Ae. aegypti. In addition, qRT-PCR results have shown the dynamics of Wolbachia across the developmental stages of mosquito. The absence of Wolbachia in tetracycline-treated Ae. aegypti mosquitoes evidenced by transmission electron microscopy reinforced our finding conclusively. After confirming their persistence through generations, we have designated Wolbachia from Ae. aegypti as wAegB. In our considered view, wAegB could play a dynamic role in impeding mosquito multiplication and consequently impinging transmission of the dreadful dengue.}, } @article {pmid30867321, year = {2019}, author = {Taylor, MJ and von Geldern, TW and Ford, L and Hübner, MP and Marsh, K and Johnston, KL and Sjoberg, HT and Specht, S and Pionnier, N and Tyrer, HE and Clare, RH and Cook, DAN and Murphy, E and Steven, A and Archer, J and Bloemker, D and Lenz, F and Koschel, M and Ehrens, A and Metuge, HM and Chunda, VC and Ndongmo Chounna, PW and Njouendou, AJ and Fombad, FF and Carr, R and Morton, HE and Aljayyoussi, G and Hoerauf, A and Wanji, S and Kempf, DJ and Turner, JD and Ward, SA}, title = {Preclinical development of an oral anti-Wolbachia macrolide drug for the treatment of lymphatic filariasis and onchocerciasis.}, journal = {Science translational medicine}, volume = {11}, number = {483}, pages = {}, doi = {10.1126/scitranslmed.aau2086}, pmid = {30867321}, issn = {1946-6242}, support = {MC_PC_16052/MRC_/Medical Research Council/United Kingdom ; MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; MR/R025401/1/MRC_/Medical Research Council/United Kingdom ; NC/M00175X/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; }, mesh = {Administration, Oral ; Animals ; Disease Models, Animal ; Elephantiasis, Filarial/blood/*drug therapy/*microbiology ; Female ; Macrolides/*administration & dosage/adverse effects/*therapeutic use ; Male ; Mice, Inbred BALB C ; Mice, SCID ; Onchocerciasis/blood/*drug therapy/*microbiology ; Treatment Outcome ; Tylosin/blood/chemical synthesis/chemistry/therapeutic use ; Wolbachia/*physiology ; }, abstract = {There is an urgent global need for a safe macrofilaricide drug to accelerate elimination of the neglected tropical diseases onchocerciasis and lymphatic filariasis. From an anti-infective compound library, the macrolide veterinary antibiotic, tylosin A, was identified as a hit against Wolbachia This bacterial endosymbiont is required for filarial worm viability and fertility and is a validated target for macrofilaricidal drugs. Medicinal chemistry was undertaken to develop tylosin A analogs with improved oral bioavailability. Two analogs, A-1535469 and A-1574083, were selected. Their efficacy was tested against the gold-standard second-generation tetracycline antibiotics, doxycycline and minocycline, in mouse and gerbil infection models of lymphatic filariasis (Brugia malayi and Litomosoides sigmodontis) and onchocerciasis (Onchocerca ochengi). A 1- or 2-week course of oral A-1535469 or A-1574083 provided >90% Wolbachia depletion from nematodes in infected animals, resulting in a block in embryogenesis and depletion of microfilarial worm loads. The two analogs delivered comparative or superior efficacy compared to a 3- to 4-week course of doxycycline or minocycline. A-1574083 (now called ABBV-4083) was selected for further preclinical testing. Cardiovascular studies in dogs and toxicology studies in rats and dogs revealed no adverse effects at doses (50 mg/kg) that achieved plasma concentrations >10-fold above the efficacious concentration. A-1574083 (ABBV-4083) shows potential as an anti-Wolbachia macrolide with an efficacy, pharmacology, and safety profile that is compatible with a short-term oral drug course for treating lymphatic filariasis and onchocerciasis.}, } @article {pmid30860942, year = {2019}, author = {Wang, J and Zhu, X and Ying, Z and Han, Q and Liao, C and Wang, J and Zhao, J and Sun, J and Lindsay, DS}, title = {Prevalence of Dirofilaria immitis Infections In Dogs and Cats In Hainan Island/Province and Three Other Coastal Cities of China Based On Antigen Testing and PCR.}, journal = {The Journal of parasitology}, volume = {105}, number = {2}, pages = {199-202}, pmid = {30860942}, issn = {1937-2345}, mesh = {Animals ; Cat Diseases/*epidemiology/parasitology ; Cats ; China/epidemiology ; Dirofilariasis/*epidemiology ; Dog Diseases/*epidemiology/parasitology ; Dogs ; Prevalence ; Urban Population ; }, abstract = {Canine and feline heartworm disease caused by Dirofilaria immitis is a serious and sometimes fatal infection transmitted by mosquitos. Little is known about the prevalence or distribution of D. immitis infection in dogs and cats on the island of Hainan island/province or coastal cities of China. The present study examined the occurrence of D. immitis infections in dogs (n = 869) and cats (n = 51) in Hainan island/province and prevalence in dogs from 3 coastal cities (Shenzhen [n = 55], Shanghai [n = 69], and Hangzhou [n = 45]) in southern and eastern China. A commercial antigen detection (AD) test and 2 PCR methods (16S ribosomal RNA and Wolbachia surface protein [ WSP] genes) were used to determine the prevalence of D. immitis from animals >6 mo old with no previous history of D. immitis preventive treatment or heartworm infection. Gene sequencing was used to confirm positive PCR samples. The AD test was not used on cat samples. Using the AD test, the prevalence in dogs was 0.5% (4/869) in Hainan island/province, 0% (0/55) in Shenzhen, 1.5% (1/69) in Shanghai, and 0% (0/45) in Hangzhou. Prevalence by 16S rRNA gene PCR was 7.4% (64/869) of dogs from Hainan island/province, 0% (0/55) in dogs from Shenzhen, 1.5% (1/69) in dogs from Shanghai, and 0% (0/45) in dogs from Hangzhou. Prevalence by WSP gene PCR in dogs was 5.3% (46/869) in Hainan island/province, 0% (0/55) in Shenzhen, 1.5% (1/69) in Shanghai, and 0% (0/45) in Hangzhou. Prevalence in the 51 cats from Hainan island/province was 9.8% and 5.9% by 16S rRNA and WSP gene PCR, respectively. The present study demonstrates that canine heartworm exposure occurs in dogs and cats in Hainan island/province and that PCR methods detected a higher prevalence than did the AD method. The 16S rRNA gene PCR detected more positive samples than did the WSP gene PCR in both dogs and cats. The 3 coastal cities had very few dogs that had evidence of D. immitis exposure.}, } @article {pmid30857919, year = {2019}, author = {Brinker, P and Fontaine, MC and Beukeboom, LW and Falcao Salles, J}, title = {Host, Symbionts, and the Microbiome: The Missing Tripartite Interaction.}, journal = {Trends in microbiology}, volume = {27}, number = {6}, pages = {480-488}, doi = {10.1016/j.tim.2019.02.002}, pmid = {30857919}, issn = {1878-4380}, mesh = {Animals ; Environment ; *Host-Pathogen Interactions ; Humans ; Microbial Interactions ; *Microbiota ; *Symbiosis ; }, abstract = {Symbiosis between microbial associates and a host is a ubiquitous feature of life on earth, modulating host phenotypes. In addition to endosymbionts, organisms harbour a collection of host-associated microbes, the microbiome that can impact important host traits. In this opinion article we argue that the mutual influences of the microbiome and endosymbionts, as well as their combined influence on the host, are still understudied. Focusing on the endosymbiont Wolbachia, we present growing evidence indicating that host phenotypic effects are exerted in interaction with the remainder microbiome and the host. We thus advocate that only through an integrated approach that considers multiple interacting partners and environmental influences will we be able to gain a better understanding of host-microbe associations.}, } @article {pmid30847475, year = {2019}, author = {Davydov, II and Salamin, N and Robinson-Rechavi, M}, title = {Large-Scale Comparative Analysis of Codon Models Accounting for Protein and Nucleotide Selection.}, journal = {Molecular biology and evolution}, volume = {36}, number = {6}, pages = {1316-1332}, pmid = {30847475}, issn = {1537-1719}, mesh = {Animals ; *Codon ; Computer Simulation ; Drosophila/genetics ; *Models, Genetic ; *Mutation Rate ; Recombination, Genetic ; *Selection, Genetic ; *Silent Mutation ; Vertebrates/genetics ; }, abstract = {There are numerous sources of variation in the rate of synonymous substitutions inside genes, such as direct selection on the nucleotide sequence, or mutation rate variation. Yet scans for positive selection rely on codon models which incorporate an assumption of effectively neutral synonymous substitution rate, constant between sites of each gene. Here we perform a large-scale comparison of approaches which incorporate codon substitution rate variation and propose our own simple yet effective modification of existing models. We find strong effects of substitution rate variation on positive selection inference. More than 70% of the genes detected by the classical branch-site model are presumably false positives caused by the incorrect assumption of uniform synonymous substitution rate. We propose a new model which is strongly favored by the data while remaining computationally tractable. With the new model we can capture signatures of nucleotide level selection acting on translation initiation and on splicing sites within the coding region. Finally, we show that rate variation is highest in the highly recombining regions, and we propose that recombination and mutation rate variation, such as high CpG mutation rate, are the two main sources of nucleotide rate variation. Although we detect fewer genes under positive selection in Drosophila than without rate variation, the genes which we detect contain a stronger signal of adaptation of dynein, which could be associated with Wolbachia infection. We provide software to perform positive selection analysis using the new model.}, } @article {pmid30841858, year = {2019}, author = {Idro, R and Anguzu, R and Ogwang, R and Akun, P and Abbo, C and Mwaka, AD and Opar, B and Nakamya, P and Taylor, M and Elliott, A and Vincent, A and Newton, C and Marsh, K}, title = {Doxycycline for the treatment of nodding syndrome (DONS); the study protocol of a phase II randomised controlled trial.}, journal = {BMC neurology}, volume = {19}, number = {1}, pages = {35}, pmid = {30841858}, issn = {1471-2377}, support = {MC_UU_00027/5/MRC_/Medical Research Council/United Kingdom ; MR/M025489/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Antiparasitic Agents/*therapeutic use ; Child ; Double-Blind Method ; Doxycycline/*therapeutic use ; Humans ; Male ; Nodding Syndrome/*drug therapy ; Onchocerca volvulus/immunology ; Quality of Life ; Research Design ; Treatment Outcome ; Uganda ; }, abstract = {BACKGROUND: Nodding syndrome is a poorly understood neurological disorder of unknown aetiology, affecting several thousand children in Africa. There has been a consistent epidemiological association with infection by the filarial parasite, Onchocerca volvulus and antibodies to leiomodin and DJ-1, cross-reacting with O.volvulus proteins, have been reported. We hypothesized that nodding syndrome is a neuro-inflammatory disorder, induced by antibodies to O.volvulus or its symbiont, Wolbachia, cross-reacting with human neuron proteins and that doxycycline, which kills Onchocerca through effects on Wolbachia, may be used as treatment.

METHODS: This will be a two-arm, double-blind, placebo-controlled, randomised phase II trial of doxycycline 100 mg daily for six weeks in 230 participants. Participants will be patients' ages≥8 years with nodding syndrome. They will receive standard of care supportive treatment. All will be hospitalised for 1-2 weeks during which time baseline measurements including clinical assessments, EEG, cognitive and laboratory testing will be performed and antiepileptic drug doses rationalised. Participants will then be randomised to either oral doxycycline (Azudox®, Kampala Pharmaceutical Industries) 100 mg daily or placebo. Treatment will be initiated in hospital and continued at home. Participants will be visited at home at 2, 4 and 6 weeks for adherence monitoring. Study outcomes will be assessed at 6, 12, 18 and 24-month visits. Analysis will be by intention to treat. The primary efficacy outcome measure will be the proportion of patients testing positive and the levels or titires of antibodies to host neuron proteins (HNPs) and/or leiomodin at 24 months. Secondary outcome measures will include effect of the intervention on seizure control, inflammatory markers, cognitive function, disease severity and quality of life.

DISCUSSION: This trial postulates that targeting O.volvulus through drugs which kill Wolbachia can modify the pathogenic processes in nodding syndrome and improve outcomes. Findings from this study are expected to substantially improve the understanding and treatment of nodding syndrome.

TRIAL REGISTRATION: Registered with clinicaltrials.gov ID: NCT02850913 on 1st August, 2016.}, } @article {pmid30841606, year = {2019}, author = {Kwon, M and Seo, SS and Kim, MK and Lee, DO and Lim, MC}, title = {Compositional and Functional Differences between Microbiota and Cervical Carcinogenesis as Identified by Shotgun Metagenomic Sequencing.}, journal = {Cancers}, volume = {11}, number = {3}, pages = {}, pmid = {30841606}, issn = {2072-6694}, support = {1610210//National Cancer Center/Republic of Korea ; }, abstract = {Recent studies have reported the potential role of microbiomes in cervical disease. However, little is known about the microbiome composition and function in cervical carcinogenesis. We aimed to identify the compositional and functional alterations of cervical microbiomes in cases of cervical carcinogenesis of Korean women using shotgun metagenomic sequencing. In this study, using shotgun sequencing, we sequenced the cervical metagenomes of cervical intraneoplasia 2/3 (n = 17), cervical cancer (n = 12), and normal controls (n = 18) to identify the microbial abundances and enriched metabolic functions in cervical metagenomes. At the genus level, the microbiota of cervical cancer were differentially enriched with genera Alkaliphilus, Pseudothermotoga, and Wolbachia. Cervical intraepithelial neoplasia (CIN) 2/3 were enriched with Lactobacillus, Staphylococcus, and Candidatus Endolissoclinum. The normal group was enriched with Pseudoalteromonas and Psychrobacter. Further characterization of the functionalities of the metagenomes may suggest that six Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologies (KOs) that are involved in 10 pathways are associated with an increased risk of CIN2/3 and cervical cancer. Specifically, cervical metagenomes were enriched in the course of peptidoglycan synthesis and depleted by dioxin degradation and 4-oxalocrotonate tautomerase. The Cluster of Orthologous Groups (COG) category 'Defense mechanisms' was depleted in cervical cancer patients. Our findings based on shotgun metagenomic sequencing suggest that cervical microbiome community compositions and their metagenomics profiles differed between cervical lesions and normal subjects. Future studies should have larger sample sizes and/or aggregate their results to have sufficient power to detect reproducible and significant associations.}, } @article {pmid30837458, year = {2019}, author = {Reveillaud, J and Bordenstein, SR and Cruaud, C and Shaiber, A and Esen, ÖC and Weill, M and Makoundou, P and Lolans, K and Watson, AR and Rakotoarivony, I and Bordenstein, SR and Eren, AM}, title = {The Wolbachia mobilome in Culex pipiens includes a putative plasmid.}, journal = {Nature communications}, volume = {10}, number = {1}, pages = {1051}, pmid = {30837458}, issn = {2041-1723}, support = {P30 DK058404/DK/NIDDK NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R21 AI133522/AI/NIAID NIH HHS/United States ; T32 EB009412/EB/NIBIB NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/genetics ; Culex/*microbiology ; Female ; France ; Genome, Bacterial/*genetics ; Host Microbial Interactions/genetics ; Metagenome/*genetics ; Metagenomics/methods ; Mosquito Vectors/microbiology ; Ovary/microbiology ; Plasmids/*genetics ; Sequence Analysis, DNA ; Symbiosis/genetics ; Wolbachia/*genetics/virology ; }, abstract = {Wolbachia is a genus of obligate intracellular bacteria found in nematodes and arthropods worldwide, including insect vectors that transmit dengue, West Nile, and Zika viruses. Wolbachia's unique ability to alter host reproductive behavior through its temperate bacteriophage WO has enabled the development of new vector control strategies. However, our understanding of Wolbachia's mobilome beyond its bacteriophages is incomplete. Here, we reconstruct near-complete Wolbachia genomes from individual ovary metagenomes of four wild Culex pipiens mosquitoes captured in France. In addition to viral genes missing from the Wolbachia reference genome, we identify a putative plasmid (pWCP), consisting of a 9.23-kbp circular element with 14 genes. We validate its presence in additional Culex pipiens mosquitoes using PCR, long-read sequencing, and screening of existing metagenomes. The discovery of this previously unrecognized extrachromosomal element opens additional possibilities for genetic manipulation of Wolbachia.}, } @article {pmid30834147, year = {2019}, author = {Pagel, L and Bultman, T and Górzyńska, K and Lembicz, M and Leuchtmann, A and Sangliana, A and Richards, N}, title = {Botanophila flies, vectors of Epichloë fungal spores, are infected by Wolbachia.}, journal = {Mycology}, volume = {10}, number = {1}, pages = {1-5}, pmid = {30834147}, issn = {2150-1203}, abstract = {Epichloë fungi are endophytes within grasses that can form stromata on culms of their hosts. Botanophila flies visit the stromata for egg laying and in the process can vector spermatial spores, thereby cross fertilising the fungus. Following egg hatch, larval flies consume fungal tissue and spores. Thus, Epichloë individuals with traits that limit larval consumption could be at a selective advantage. We assessed Botanophila fly larvae from sites within the United States and Europe for infection by the bacterial sexual parasite Wolbachia through amplification of the Wolbachia surface protein gene (wsp). Nearly 70% of fly larvae in our samples were infected by Wolbachia. This is the first record of infection by Wolbachia within Botanophila and could have far reaching effects on not only the fly host, but also the Epichloë fungi upon which Botanophila feeds as well as the grass host within which the fungi live. For example, infection by Wolbachia could limit consumption of Epichloë spores by Botanophila larvae if the bacteria promoted premature larval death.}, } @article {pmid30820826, year = {2019}, author = {Onder, Z and Ciloglu, A and Duzlu, O and Yildirim, A and Okur, M and Yetismis, G and Inci, A}, title = {Molecular detection and identification of Wolbachia endosymbiont in fleas (Insecta: Siphonaptera).}, journal = {Folia microbiologica}, volume = {64}, number = {6}, pages = {789-796}, pmid = {30820826}, issn = {1874-9356}, mesh = {Animals ; DNA, Bacterial/genetics ; Female ; Flea Infestations/epidemiology/parasitology/*veterinary ; Genes, Bacterial/genetics ; Genetic Variation ; Genome, Bacterial/genetics ; Host Specificity ; Male ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Siphonaptera/*microbiology ; Symbiosis ; Turkey/epidemiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {The aim of this study was to determine the presence and prevalence of Wolbachia bacteria in natural population of fleas (Insecta: Siphonaptera) in Turkey, and to exhibit the molecular characterization and the phylogenetic reconstruction at the positive isolates with other species in GenBank, based on 16S rDNA sequences. One hundred twenty-four flea samples belonging to the species Ctenocephalides canis, C. felis, and Pulex irritans were collected from animal shelters in Kayseri between January and August 2017. All flea species were individually screened for the presence of Wolbachia spp. by polymerase chain reaction (PCR) targeting the 16S ribosomal RNA gene. According to PCR analyses, Wolbachia spp. were found prevalent in C. canis and P. irritans fleas, while it was not detected in the C. felis species. Totally, 20 isolates were purified from agarose gel and sequenced with the same primers for molecular characterization and phylogenetic analyses. The sequence analyses revealed 17 polymorphic sites and 2 genetically different Wolbachia isolates, representing two different haplotypes in two flea species. The distribution patterns, molecular characterization, and phylogenetic status of Wolbachia spp. of fleas in Turkey are presented for the first time with this study. Understanding of the role of Wolbachia in vector biology may provide information for developing Wolbachia-based biological control tools.}, } @article {pmid30819117, year = {2019}, author = {Fortin, M and Meunier, J and Laverré, T and Souty-Grosset, C and Richard, FJ}, title = {Joint effects of group sex-ratio and Wolbachia infection on female reproductive success in the terrestrial isopod Armadillidium vulgare.}, journal = {BMC evolutionary biology}, volume = {19}, number = {1}, pages = {65}, pmid = {30819117}, issn = {1471-2148}, mesh = {Animals ; Female ; Fertility ; Isopoda/microbiology/*physiology ; Male ; Phenotype ; Reproduction ; *Sex Ratio ; *Sexual Behavior, Animal ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: In species that reproduce with sexual reproduction, males and females often have opposite strategies to maximize their own fitness. For instance, males are typically expected to maximize their number of mating events, whereas an excessive number of mating events can be costly for females. Although the risk of sexual harassment by males and resulting costs for females are expected to increase with the proportion of males, it remains unknown whether and how parasitic distorters of a host population's sex-ratio can shape this effect on the fitness of females. Here, we addressed this question using Armadillidium vulgare and its parasite Wolbachia that alters the sex-ratio of a population. We set up Wolbachia-free and Wolbachia-infected females in experimental groups exhibiting 100, 80, 50% or 20% females for 1 year, during which we measured changes in survival, fertility and fecundity.

RESULTS: Wolbachia infection shaped the effects of both population sex-ratio and reproductive season on female fecundity. Compared to Wolbachia-free females, Wolbachia-infected females were less likely to be gravid in populations exhibiting an excess of females and did not exhibit the otherwise negative effect of seasons on this likelihood. Group sex-ratio and Wolbachia infection have independent effects on other measured traits. Male-biased populations had females both exhibiting the lowest survival rate after 6 months and producing the smallest number of offspring, independent of Wolbachia infection. Conversely, Wolbachia-infected females had the lowest likelihood of producing at least one offspring, independent of group sex-ratio. Wolbachia infection had no effect on female survival rate.

CONCLUSIONS: We demonstrated that male-biased sex-ratio and the presence of Wolbachia are costly for females due to sexual harassment by males and bacterial infection, respectively. Interestingly, Wolbachia infection triggers another negative effect. This effect does not come from an excess of males and its associated sexual harassment of females but instead from a lack of males and the associated risk for females of remaining unmated. Overall, these findings highlight the importance of social pressures and infection on female fitness and provide insights into our general understanding of the joint and opposite effects of these two parameters in the evolution of reproductive strategies.}, } @article {pmid30818326, year = {2019}, author = {von Geldern, TW and Morton, HE and Clark, RF and Brown, BS and Johnston, KL and Ford, L and Specht, S and Carr, RA and Stolarik, DF and Ma, J and Rieser, MJ and Struever, D and Frohberger, SJ and Koschel, M and Ehrens, A and Turner, JD and Hübner, MP and Hoerauf, A and Taylor, MJ and Ward, SA and Marsh, K and Kempf, DJ}, title = {Discovery of ABBV-4083, a novel analog of Tylosin A that has potent anti-Wolbachia and anti-filarial activity.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {2}, pages = {e0007159}, pmid = {30818326}, issn = {1935-2735}, support = {MC_PC_16052/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacokinetics/*pharmacology ; *Drug Discovery ; Elephantiasis, Filarial/drug therapy ; Female ; Filaricides/pharmacokinetics/*pharmacology ; Filarioidea/drug effects/microbiology ; Gerbillinae ; Mice ; Mice, Inbred BALB C ; Onchocerciasis/drug therapy ; Symbiosis/drug effects ; Tylosin/*analogs & derivatives/*pharmacology ; Wolbachia/*drug effects ; }, abstract = {There is a significant need for improved treatments for onchocerciasis and lymphatic filariasis, diseases caused by filarial worm infection. In particular, an agent able to selectively kill adult worms (macrofilaricide) would be expected to substantially augment the benefits of mass drug administration (MDA) with current microfilaricides, and to provide a solution to treatment of onchocerciasis / loiasis co-infection, where MDA is restricted. We have identified a novel macrofilaricidal agent, Tylosin A (TylA), which acts by targeting the worm-symbiont Wolbachia bacterium. Chemical modification of TylA leads to improvements in anti-Wolbachia activity and oral pharmacokinetic properties; an optimized analog (ABBV-4083) has been selected for clinical evaluation.}, } @article {pmid30817823, year = {2019}, author = {Staunton, KM and Yeeles, P and Townsend, M and Nowrouzi, S and Paton, CJ and Trewin, B and Pagendam, D and Bondarenco, A and Devine, GJ and Snoad, N and Beebe, NW and Ritchie, SA}, title = {Trap Location and Premises Condition Influences on Aedes aegypti (Diptera: Culicidae) Catches Using Biogents Sentinel Traps During a 'Rear and Release' Program: Implications for Designing Surveillance Programs.}, journal = {Journal of medical entomology}, volume = {56}, number = {4}, pages = {1102-1111}, doi = {10.1093/jme/tjz018}, pmid = {30817823}, issn = {1938-2928}, mesh = {*Aedes ; Animals ; *Environment ; Female ; Male ; *Mosquito Control ; }, abstract = {As the incidence of arboviral diseases such as dengue, Zika, chikungunya, and yellow fever increases globally, controlling their primary vector, Aedes aegypti (L.) (Diptera: Culicidae), is of greater importance than ever before. Mosquito control programs rely heavily on effective adult surveillance to ensure methodological efficacy. The Biogents Sentinel (BGS) trap is the gold standard for surveilling adult Aedes mosquitoes and is commonly deployed worldwide, including during modern 'rear and release' programs. Despite its extensive use, few studies have directly assessed environmental characteristics that affect BGS trap catches, let alone how these influences change during 'rear and release' programs. We assessed male and female Ae. aegypti spatial stability, as well as premises condition and trap location influences on BGS trap catches, as part of Debug Innisfail 'rear and release' program in northern Australia. We found similar trends in spatial stability of male and female mosquitoes at both weekly and monthly resolutions. From surveillance in locations where no males were released, reduced catches were found at premises that contained somewhat damaged houses and unscreened properties. In addition, when traps were located in areas that were unsheltered, more than 10 m from commonly used sitting areas or more visually complex catches were also negatively affected. In locations where males were released, we found that traps in treatment sites, relative to control sites, displayed increased catches in heavily shaded premises and were inconsistently influenced by differences in house sets and building materials. Such findings have valuable implications for a range of Ae. aegypti surveillance programs.}, } @article {pmid30813886, year = {2019}, author = {Bykov, RА and Yudina, MA and Gruntenko, NE and Zakharov, IK and Voloshina, MA and Melashchenko, ES and Danilova, MV and Mazunin, IO and Ilinsky, YY}, title = {Prevalence and genetic diversity of Wolbachia endosymbiont and mtDNA in Palearctic populations of Drosophila melanogaster.}, journal = {BMC evolutionary biology}, volume = {19}, number = {Suppl 1}, pages = {48}, pmid = {30813886}, issn = {1471-2148}, support = {F32 MH010832/MH/NIMH NIH HHS/United States ; }, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Drosophila melanogaster/*microbiology ; *Genetic Variation ; Genomics ; Geography ; Haplotypes ; Mitochondria/genetics ; Prevalence ; *Symbiosis ; Wolbachia/cytology/*genetics/*physiology ; }, abstract = {BACKGROUND: Maternally inherited Wolbachia symbionts infect D. melanogaster populations worldwide. Infection rates vary greatly. Genetic diversity of Wolbachia in D. melanogaster can be subdivided into several closely related genotypes coinherited with certain mtDNA lineages. mtDNA haplotypes have the following global distribution pattern: mtDNA clade I is mostly found in North America, II and IV in Africa, III in Europe and Africa, V in Eurasia, VI is global but very rare, and VIII is found in Asia. The wMel Wolbachia genotype is predominant in D. melanogaster populations. However, according to the hypothesis of global Wolbachia replacement, the wMelCS genotype was predominant before the XX century when it was replaced by the wMel genotype. Here we analyse over 1500 fly isolates from the Palearctic region to evaluate the prevalence, genetic diversity and distribution pattrern of the Wolbachia symbiont, occurrence of mtDNA variants, and finally to discuss the Wolbachia genotype global replacement hypothesis.

RESULTS: All studied Palearctic populations of D. melanogaster were infected with Wolbachia at a rate of 33-100%. We did not observe any significant correlation between infection rate and longitude or latitude. Five previously reported Wolbachia genotypes were found in Palearctic populations with a predominance of the wMel variant. The mtDNA haplotypes of the I_II_III clade and V clade were prevalent in Palearctic populations. To test the recent Wolbachia genotype replacement hypothesis, we examined three genomic regions of CS-like genotypes. Low genetic diversity was observed, only two haplotypes of the CS genotypes with a 'CCG' variant predominance were found.

CONCLUSION: The results of our survey of Wolbachia infection prevalence and genotype diversity in Palearctic D. melanogaster populations confirm previous studies. Wolbachia is ubiquitous in the Palearctic region. The wMel genotype is dominant with local occurrence of rare genotypes. Together with variants of the V mtDNA clade, the variants of the 'III+' clade are dominant in both infected and uninfected flies of Palearctic populations. Based on our data on Wolbachia and mtDNA in different years in some Palearctic localities, we can conclude that flies that survive the winter make the predominant symbiont contribution to the subsequent generation. A comprehensive overview of mtDNA and Wolbachia infection of D. melanogaster populations worldwide does not support the recent global Wolbachia genotype replacement hypothesis. However, we cannot exclude wMelCS genotype rate fluctuations in the past.}, } @article {pmid30810656, year = {2019}, author = {Cândido, EL and Silva, UMD and Cavalcanti, LPG}, title = {New record and extended geographical distribution of Aedes fluviatilis (Lutz, 1904) in Ceará, northeastern Brazil.}, journal = {Revista da Sociedade Brasileira de Medicina Tropical}, volume = {52}, number = {}, pages = {e20180286}, doi = {10.1590/0037-8682-0286-2018}, pmid = {30810656}, issn = {1678-9849}, mesh = {Aedes/*classification ; Animals ; Brazil ; Ecosystem ; Mosquito Vectors/*classification ; Spatial Analysis ; }, abstract = {INTRODUCTION: Aedes fluviatilis(Lutz, 1904) is considered a potential vector of yellow fever and dengue viruses and is naturally infected by Wolbachia.

METHODS: In March 2018, during a field activity in the municipality of Saboeiro, 163 mosquito larvae were collected in a dammed area of the Jaguaribe River.

RESULTS: Of the larvae collected, 143 (87.7%) were identified asAe. fluviatilis.

CONCLUSIONS: We report the first documentation of Ae. fluviatilis in the municipality of Saboeiro, northeastern Brazil. It is important to conduct additional entomological surveys to characterize the local Culicidae fauna. Ignoring the presence and dispersion of this vector could be a public health risk.}, } @article {pmid30807733, year = {2019}, author = {Bagheri, Z and Talebi, AA and Asgari, S and Mehrabadi, M}, title = {Wolbachia induce cytoplasmic incompatibility and affect mate preference in Habrobracon hebetor to increase the chance of its transmission to the next generation.}, journal = {Journal of invertebrate pathology}, volume = {163}, number = {}, pages = {1-7}, doi = {10.1016/j.jip.2019.02.005}, pmid = {30807733}, issn = {1096-0805}, mesh = {Animals ; Biological Control Agents ; Female ; Fertility ; *Host Microbial Interactions ; *Infectious Disease Transmission, Vertical ; Male ; Sex Ratio ; Sexual Behavior, Animal ; Symbiosis ; Wasps/*microbiology ; *Wolbachia/isolation & purification ; }, abstract = {Wolbachia are common intracellular bacteria that are generally found in arthropods, including a high proportion of insects and also some nematodes. This intracellular symbiont can affect sex ratio with a variety of reproductive anomalies in the host, including cytoplasmic incompatibility (CI) in haplodiploids. In this study, we questioned if the parasitoid wasp, Habrobracon hebetor (Hym.: Braconidae), an important biological control agent of many lepidopteran larvae, is infected with Wolbachia. To test this, DNA was extracted from adult insects and subjected to PCR using specific primers to Wolbachia target genes. The results showed a high rate of Wolbachia infection in this parasitoid wasp. To determine the biological function of Wolbachia in H. hebetor, we removed this bacterium from the wasps using antibiotic treatment (cured wasps). Results of crossing experiments revealed that Wolbachia induced CI in H. hebetor in which cured females crossed with infected males produced only males, while both male and female progeny were observed for other crosses. Also, we showed that the presence of Wolbachia in females increased fecundity and female offspring of this parasitoid wasp. The presence of Wolbachia in the males had no significant effect on fecundity and female production, but might have incurred costs. We also investigated the effect of Wolbachia on mate choice and found that Wolbachia affects mating behavior of H. hebetor. Together, we showed that Wolbachia induces CI in H. hebetor and affects host mating behavior in favor of its transmission. Wolbachia utilize these strategies to increase the frequency of infected females in the host population.}, } @article {pmid30798391, year = {2019}, author = {Singhal, K and Mohanty, S}, title = {Genome organisation and comparative genomics of four novel Wolbachia genome assemblies from Indian Drosophila host.}, journal = {Functional & integrative genomics}, volume = {19}, number = {4}, pages = {617-632}, pmid = {30798391}, issn = {1438-7948}, mesh = {Animals ; Bacterial Proteins/genetics/metabolism ; Drosophila/microbiology ; *Genome, Bacterial ; Host-Pathogen Interactions ; *Polymorphism, Genetic ; Symbiosis ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia has long been known to share an endosymbiotic relationship with its host as an obligate intracellular organism. Wolbachia diversity as different supergroups is found to be host-specific in most cases except a few, where the host species is seen to accommodate multiple strains. Besides, the Wolbachia genome must have undergone several changes in response to the evolving host genome in order to adapt and establish a strong association with its host, thus making a distinctive Wolbachia-host alliance. The present study focusses on four novel genome assembly and genome-wide sequence variations of Indian Wolbachia strains, i.e. wMel and wRi isolated from two different Drosophila hosts. The genome assembly has an average size of ~ 1.1 Mb and contains ~ 1100 genes, which is comparable with the previously sequenced Wolbachia genomes. The comparative genomics analysis of these genomes and sequence-wide comparison of some functionally significant genes, i.e. ankyrin repeats, Wsp and T4SS, highlight their sequence similarities and dissimilarities, further supporting the strain-specific association of Wolbachia to its host. Interestingly, some of the sequence variations are also found to be restricted to only Indian Wolbachia strains. Further analysis of prophage and their flanking regions in the Wolbachia genome reveals the presence of several functional genes which may assist the phage to reside inside the bacterial host, thus providing a trade-off for the endosymbiont-host association. Understanding this endosymbiont genome in different eco-geographical conditions has become imperative for the recent use of Wolbachia in medical entomology as a vector-control agent.}, } @article {pmid30796064, year = {2019}, author = {Newton, ILG and Slatko, BE}, title = {Symbiosis Comes of age at the 10[th] Biennial Meeting of Wolbachia Researchers.}, journal = {Applied and environmental microbiology}, volume = {85}, number = {8}, pages = {}, pmid = {30796064}, issn = {1098-5336}, abstract = {Wolbachia pipientis is an alpha-proteobacterial, obligate intracellular microbe and arguably the most successful infection on our planet, colonizing 40-60% of insect species. Wolbachia are also present in most, but not all, filarial nematodes where they are obligate mutualists and are the targets for anti-filarial drug discovery. Although Wolbachia are related to important human pathogens they do not infect mammals, but instead are well known for their reproductive manipulations of insect populations, inducing the following phenotypes: male-killing, feminization, parthenogenesis induction, or cytoplasmic incompatibility (CI). The most common of these, CI, results in a sperm-egg incompatibility and increases the relative fecundity of infected females in a population. In the last decade, Wolbachia have also been shown to provide a benefit to insects, where the infection can inhibit RNA virus replication within the host. Wolbachia cannot be cultivated outside of host cells and no genetic tools are available in the symbiont, limiting approaches available to its study. This means that many questions fundamental to our understanding of Wolbachia basic biology remained unknown for decades. The tenth biennial international Wolbachia conference, "Wolbachia Evolution, Ecology, Genomics and Cell Biology: A Chronicle of the Most Ubiquitous Symbiont", was held on June 17-22, 2018, Salem, MA USA. In the review below we highlight the new science presented at the meeting, link it to prior efforts to answer these questions across the Wolbachia genus, and the importance to the field of symbiosis. The topics covered in this review are based on the presentations at the conference.}, } @article {pmid30762095, year = {2019}, author = {Martínez-Rodríguez, P and Rolán-Alvarez, E and Del Mar Pérez-Ruiz, M and Arroyo-Yebras, F and Carpena-Catoira, C and Carvajal-Rodríguez, A and Bella, JL}, title = {Geographic and Temporal Variation of Distinct Intracellular Endosymbiont Strains of Wolbachia sp. in the Grasshopper Chorthippus parallelus: a Frequency-Dependent Mechanism?.}, journal = {Microbial ecology}, volume = {77}, number = {4}, pages = {1036-1047}, pmid = {30762095}, issn = {1432-184X}, support = {CGL2016-75482-P//Ministerio de Ciencia y Tecnolog?a/ ; BFU2013-44635//Ministerio de Econom?a, Industria y Competitividad, Gobierno de Espa?a/ ; }, mesh = {Animals ; Biological Coevolution ; Computer Simulation ; Geography ; Grasshoppers/*microbiology ; Linear Models ; *Polymorphism, Genetic ; Seasons ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an intracellular endosymbiont that can produce a range of effects on host fitness, but the temporal dynamics of Wolbachia strains have rarely been experimentally evaluated. We compare interannual strain frequencies along a geographical region for understanding the forces that shape Wolbachia strain frequency in natural populations of its host, Chorthippus parallelus (Orthoptera, Acrididae). General linear models show that strain frequency changes significantly across geographical and temporal scales. Computer simulation allows to reject the compatibility of the observed patterns with either genetic drift or sampling errors. We use consecutive years to estimate total Wolbachia strain fitness. Our estimation of Wolbachia fitness is significant in most cases, within locality and between consecutive years, following a negatively frequency-dependent trend. Wolbachia spp. B and F strains show a temporal pattern of variation that is compatible with a negative frequency-dependent natural selection mechanism. Our results suggest that such a mechanism should be at least considered in future experimental and theoretical research strategies that attempt to understand Wolbachia biodiversity.}, } @article {pmid30761981, year = {2019}, author = {Showler, AJ and Kubofcik, J and Ricciardi, A and Nutman, TB}, title = {Differences in the Clinical and Laboratory Features of Imported Onchocerciasis in Endemic Individuals and Temporary Residents.}, journal = {The American journal of tropical medicine and hygiene}, volume = {100}, number = {5}, pages = {1216-1222}, pmid = {30761981}, issn = {1476-1645}, mesh = {Acute Disease ; Adult ; Animals ; Antibodies, Helminth/blood ; Asymptomatic Infections ; Chronic Disease ; Clinical Laboratory Techniques/*statistics & numerical data ; *Emigration and Immigration ; Endemic Diseases/*statistics & numerical data ; Eye Diseases/diagnosis/parasitology ; Female ; Humans ; Immunoglobulin G/blood ; Male ; Middle Aged ; Onchocerca volvulus ; Onchocerciasis/blood/*diagnosis ; }, abstract = {Many parasitic infections have different presenting features in endemic individuals (ENDs) and immunologically naive temporary residents (TRs). Temporary residents with loiasis often display acute symptoms and hypereosinophilia, in contrast to a parasite-induced subclinical state in chronically infected ENDs. Few studies have examined differences in ENDs and TRs infected with the related filarial parasite Onchocerca volvulus. We identified 40 TRs and 36 ENDs with imported onchocerciasis at the National Institutes of Health between 1976 and 2016. All study subjects received an extensive pretreatment medical history, physical examination, and laboratory investigations. We performed additional parasite-specific serologic testing on stored patient sera. Asymptomatic infection occurred in 12.5% of TRs and no ENDs (P = 0.06). Papular dermatitis was more common in TRs (47.5% versus 2.7%, P < 0.001), whereas more pigmentation changes occurred in ENDs (41.7% versus 15%, P = 0.01). Only endemic patients reported visual disturbance (13% versus 0%, P = 0.03). One TR (3.3%) had onchocercal eye disease, compared with 22.6% of ENDs (P = 0.053). Absolute eosinophil counts (AECs) were similar in ENDs and TRs (P = 0.5), and one-third of subjects had a normal AEC. Endemic individuals had higher filarial-specific IgG4 and were more likely to be positive for IgG4 antibodies to Ov-16. Temporary residents and ENDs with imported O. volvulus infection presented with different dermatologic manifestations; ocular involvement occurred almost exclusively in ENDs. Unlike Loa loa, clinical differences appear not to be eosinophil-mediated and may reflect chronicity, intensity of infection, or the presence of Wolbachia in O. volvulus.}, } @article {pmid30747079, year = {2019}, author = {Chegeni, TN and Fakhar, M}, title = {Promising Role of Wolbachia as Anti-parasitic Drug Target and Eco-Friendly Biocontrol Agent.}, journal = {Recent patents on anti-infective drug discovery}, volume = {14}, number = {1}, pages = {69-79}, doi = {10.2174/1574891X14666190211162403}, pmid = {30747079}, issn = {2212-4071}, mesh = {Animals ; Antiparasitic Agents/*pharmacology/*therapeutic use ; Humans ; Parasites/*drug effects ; Patents as Topic ; Wolbachia/*drug effects ; }, abstract = {BACKGROUND: Wolbachia is the most common endosymbiotic bacteria in insectborne parasites and it is the most common reproductive parasite in the world. Wolbachia has been found worldwide in numerous arthropod and parasite species, including insects, terrestrial isopods, spiders, mites and filarial nematodes. There is a complicated relationship between Wolbachia and its hosts and in some cases, they create a mutual relationship instead of a parasitic relationship. Some species are not able to reproduce in the absence of infection with Wolbachia. Thus, the use of existing strains of Wolbachia bacteria offers a potential strategy for the control of the population of mosquitoes and other pests and diseases.

METHODS: We searched ten databases and reviewed published papers regarding the role of Wolbachia as a promising drug target and emerging biological control agents of parasitic diseases between 1996 and 2017 (22 years) were considered eligible. Also, in the current study several patents (WO008652), (US7723062), and (US 0345249 A1) were reviewed.

RESULTS: Endosymbiotic Wolbachia bacteria, which are inherited from mothers, is transmitted to mosquitoes and interferes with pathogen transmission. They can change the reproduction of their host. Wolbachia is transmitted through the cytoplasm of eggs and have evolved different mechanisms for manipulating the reproduction of its hosts, including the induction of reproductive incompatibility, parthenogenesis, and feminization. The extensive effects of Wolbachia on reproduction and host fitness have made Wolbachia the issue of growing attention as a potential biocontrol agent.

CONCLUSION: Wolbachia has opened a new window to design a costly, potent and ecofriendly drug target for effective treatment and elimination of vector-borne parasitic diseases.}, } @article {pmid30744707, year = {2019}, author = {Augustinos, AA and Moraiti, CA and Drosopoulou, E and Kounatidis, I and Mavragani-Tsipidou, P and Bourtzis, K and Papadopoulos, NT}, title = {Old residents and new arrivals of Rhagoletis species in Europe.}, journal = {Bulletin of entomological research}, volume = {109}, number = {6}, pages = {701-712}, doi = {10.1017/S0007485319000063}, pmid = {30744707}, issn = {1475-2670}, mesh = {Animals ; Europe ; Insect Control/methods ; Introduced Species ; Population Dynamics ; Tephritidae/*classification/genetics/microbiology ; Wolbachia/physiology ; }, abstract = {The genus Rhagoletis (Diptera: Tephritidae) comprises more than 65 species distributed throughout Europe, Asia and America, including many species of high economic importance. Currently, there are three Rhagoletis species that infest fruits and nuts in Europe. The European cherry fruit fly, Rhagoletis cerasi (may have invaded Europe a long time ago from the Caucasian area of West Asia), and two invasive species (recently introduced from North America): the eastern American cherry fruit fly, R. cingulata, and the walnut husk fly, R. completa. The presence of different Rhagoletis species may enhance population dynamics and establish an unpredictable economic risk for several fruit and nut crops in Europe. Despite their excessive economic importance, little is known on population dynamics, genetics and symbiotic associations for making sound pest control decisions in terms of species-specific, environmental friendly pest control methods. To this end, the current paper (a) summarizes recently accumulated genetic and population data for the European Rhagoletis species and their association with the endosymbiont Wolbachia pipientis, and (b) explores the possibility of using the current knowledge for implementing the innovative biological control methods of sterile insect technique and incompatible insect technique.}, } @article {pmid30735918, year = {2019}, author = {Ju, H and Zhu, D and Qiao, M}, title = {Effects of polyethylene microplastics on the gut microbial community, reproduction and avoidance behaviors of the soil springtail, Folsomia candida.}, journal = {Environmental pollution (Barking, Essex : 1987)}, volume = {247}, number = {}, pages = {890-897}, doi = {10.1016/j.envpol.2019.01.097}, pmid = {30735918}, issn = {1873-6424}, mesh = {Animals ; Arthropods/drug effects/*physiology ; Avoidance Learning ; Bacteria ; Gastrointestinal Microbiome/*drug effects ; Microbiota ; Plastics/toxicity ; Polyethylene/*toxicity ; Reproduction/drug effects ; Soil ; Soil Pollutants/*toxicity ; }, abstract = {Microplastics (MPs) are an emerging contaminant and are confirmed to be ubiquitous in the environment. Adverse effects of MPs on aquatic organisms have been widely studied, whereas little research has focused on soil invertebrates. We exposed the soil springtail Folsomia candida to artificial soils contaminated with polyethylene MPs (<500 μm) for 28 d to explore the effects of MPs on avoidance, reproduction, and gut microbiota. Springtails exhibited avoidance behaviors at 0.5% and 1% MPs (w/w in dry soil), and the avoidance rate was 59% and 69%, respectively. Reproduction was inhibited when the concentration of MPs reached 0.1% and was reduced by 70.2% at the highest concentration of 1% MPs compared to control. The half-maximal effective concentration (EC50) value based on reproduction for F. candida was 0.29% MPs. At concentrations of 0.5% dry weight in the soil, MPs significantly altered the microbial community and decreased bacterial diversity in the springtail gut. Specifically, the relative abundance of Wolbachia significantly decreased while the relative abundance of Bradyrhizobiaceae, Ensifer and Stenotrophomonas significantly increased. Our results demonstrated that MPs exerted a significant toxic effect on springtails and can change their gut microbial community. This can provide useful information for risk assessment of MPs in terrestrial ecosystems.}, } @article {pmid30730745, year = {2019}, author = {Jacobs, RT and Lunde, CS and Freund, YR and Hernandez, V and Li, X and Xia, Y and Carter, DS and Berry, PW and Halladay, J and Rock, F and Stefanakis, R and Easom, E and Plattner, JJ and Ford, L and Johnston, KL and Cook, DAN and Clare, R and Cassidy, A and Myhill, L and Tyrer, H and Gamble, J and Guimaraes, AF and Steven, A and Lenz, F and Ehrens, A and Frohberger, SJ and Koschel, M and Hoerauf, A and Hübner, MP and McNamara, CW and Bakowski, MA and Turner, JD and Taylor, MJ and Ward, SA}, title = {Boron-Pleuromutilins as Anti- Wolbachia Agents with Potential for Treatment of Onchocerciasis and Lymphatic Filariasis.}, journal = {Journal of medicinal chemistry}, volume = {62}, number = {5}, pages = {2521-2540}, pmid = {30730745}, issn = {1520-4804}, mesh = {Animals ; Boron/chemistry/*pharmacology ; Diterpenes/chemistry/*pharmacology ; Elephantiasis, Filarial/*drug therapy ; Filaricides/pharmacokinetics/pharmacology/*therapeutic use ; Mice ; Mice, Inbred BALB C ; Mice, SCID ; Onchocerciasis/*drug therapy ; Polycyclic Compounds/chemistry/*pharmacology ; Wolbachia/*drug effects ; Wuchereria bancrofti/*drug effects ; Pleuromutilins ; }, abstract = {A series of pleuromutilins modified by introduction of a boron-containing heterocycle on C(14) of the polycyclic core are described. These analogs were found to be potent anti- Wolbachia antibiotics and, as such, may be useful in the treatment of filarial infections caused by Onchocerca volvulus, resulting in Onchocerciasis or river blindness, or Wuchereria bancrofti and Brugia malayi and related parasitic nematodes resulting in lymphatic filariasis. These two important neglected tropical diseases disproportionately impact patients in the developing world. The lead preclinical candidate compound containing 7-fluoro-6-oxybenzoxaborole (15, AN11251) was shown to have good in vitro anti- Wolbachia activity and physicochemical and pharmacokinetic properties providing high exposure in plasma. The lead was effective in reducing the Wolbachia load in filarial worms following oral administration to mice.}, } @article {pmid30729921, year = {2019}, author = {Simo, G and Kanté, ST and Madinga, J and Kame, G and Farikou, O and Ilombe, G and Geiger, A and Lutumba, P and Njiokou, F}, title = {Molecular identification of Wolbachia and Sodalis glossinidius in the midgut of Glossina fuscipes quanzensis from the Democratic Republic of Congo.}, journal = {Parasite (Paris, France)}, volume = {26}, number = {}, pages = {5}, pmid = {30729921}, issn = {1776-1042}, mesh = {Animals ; Coinfection/microbiology ; DNA, Bacterial/genetics ; Democratic Republic of the Congo ; Digestive System/*microbiology ; Enterobacteriaceae/*genetics/isolation & purification ; Fructose-Bisphosphate Aldolase/genetics ; High-Throughput Nucleotide Sequencing ; Insect Vectors/microbiology ; Polymerase Chain Reaction ; Symbiosis ; Tsetse Flies/*microbiology ; Wolbachia/*genetics/isolation & purification ; }, abstract = {During the last 30 years, investigations on the microbiome of different tsetse species have generated substantial data on the bacterial flora of these cyclical vectors of African trypanosomes, with the overarching goal of improving the control of trypanosomiases. It is in this context that the presence of Wolbachia and Sodalis glossinidius was studied in wild populations of Glossina fuscipes quanzensis from the Democratic Republic of Congo. Tsetse flies were captured with pyramidal traps. Of the 700 Glossina f. quanzensis captured, 360 were dissected and their midguts collected and analyzed. Sodalis glossinidius and Wolbachia were identified by PCR. The Wolbachia-positive samples were genetically characterized with five molecular markers. PCR revealed 84.78% and 15.55% midguts infected by Wolbachia and S. glossinidius, respectively. The infection rates varied according to capture sites. Of the five molecular markers used to characterize Wolbachia, only the fructose bis-phosphate aldolase gene was amplified for about 60% of midguts previously found with Wolbachia infections. The sequencing results confirmed the presence of Wolbachia and revealed the presence of S. glossinidius in the midgut of Glossina f. quanzensis. A low level of midguts were naturally co-infected by both bacteria. The data generated in this study open a framework for investigations aimed at understanding the contribution of these symbiotic microorganisms to the vectorial competence of Glossina fuscipes quanzensis.}, } @article {pmid30729319, year = {2019}, author = {Horváth, G and Garamszegi, LZ and Bereczki, J and Urszán, TJ and Balázs, G and Herczeg, G}, title = {Roll with the fear: environment and state dependence of pill bug (Armadillidium vulgare) personalities.}, journal = {Die Naturwissenschaften}, volume = {106}, number = {3-4}, pages = {7}, pmid = {30729319}, issn = {1432-1904}, support = {OTKA-K 105517//Országos Tudományos Kutatási Alapprogramok/ ; OTKA-K 109223//Országos Tudományos Kutatási Alapprogramok/ ; SNN 125627//National Research, Development and Innovation Fund for international cooperation/ ; CGL2015-70639-P//Ministerio de Economía y Competitividad/ ; K 115970//National Research, Development and Innovation Office/ ; K 129215//National Research, Development and Innovation Office/ ; }, mesh = {Animals ; Behavior, Animal/*physiology ; *Environment ; Isopoda/microbiology/*physiology ; Models, Animal ; Wolbachia/physiology ; }, abstract = {Most studies on animal personality evaluate individual mean behaviour to describe individual behavioural strategy, while often neglecting behavioural variability on the within-individual level. However, within-individual behavioural plasticity (variation induced by environment) and within-individual residual variation (regulatory behavioural precision) are recognized as biologically valid components of individual behaviour, but the evolutionary ecology of these components is still less understood. Here, we tested whether behaviour of common pill bugs (Armadillidium vulgare) differs on the among- and within-individual level and whether it is affected by various individual specific state-related traits (sex, size and Wolbachia infection). To this aim, we assayed risk-taking in familiar vs. unfamiliar environments 30 times along 38 days and applied double modelling statistical technique to handle the complex hierarchical structure for both individual-specific trait means and variances. We found that there are significant among-individual differences not only in mean risk-taking behaviour but also in environment- and time-induced behavioural plasticity and residual variation. Wolbachia-infected individuals took less risk than healthy conspecifics; in addition, individuals became more risk-averse with time. Residual variation decreased with time, and individuals expressed higher residual variation in the unfamiliar environment. Further, sensitization was stronger in females and in larger individuals in general. Our results suggest that among-individual variation, behavioural plasticity and residual variation are all (i) biologically relevant components of an individual's behavioural strategy and (ii) responsive to changes in environment or labile state variables. We propose pill bugs as promising models for personality research due to the relative ease of getting repeated behavioural measurements.}, } @article {pmid30727958, year = {2019}, author = {Kampfraath, AA and Klasson, L and Anvar, SY and Vossen, RHAM and Roelofs, D and Kraaijeveld, K and Ellers, J}, title = {Genome expansion of an obligate parthenogenesis-associated Wolbachia poses an exception to the symbiont reduction model.}, journal = {BMC genomics}, volume = {20}, number = {1}, pages = {106}, pmid = {30727958}, issn = {1471-2164}, support = {865.12.003//Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ ; 15494//Stichting voor de Technische Wetenschappen/ ; }, mesh = {Animals ; Arthropods/*microbiology/physiology ; DNA Repair ; Evolution, Molecular ; *Gene Transfer, Horizontal ; *Genome, Bacterial ; *Parthenogenesis ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {BACKGROUND: Theory predicts that dependency within host-endosymbiont interactions results in endosymbiont genome size reduction. Unexpectedly, the largest Wolbachia genome was found in the obligate, parthenogenesis-associated wFol. In this study, we investigate possible processes underlying this genome expansion by comparing a re-annotated wFol genome to other Wolbachia genomes. In addition, we also search for candidate genes related to parthenogenesis induction (PI).

RESULTS: Within wFol, we found five phage WO regions representing 25.4% of the complete genome, few pseudogenized genes, and an expansion of DNA-repair genes in comparison to other Wolbachia. These signs of genome conservation were mirrored in the wFol host, the springtail F. candida, which also had an expanded DNA-repair gene family and many horizontally transferred genes. Across all Wolbachia genomes, there was a strong correlation between gene numbers of Wolbachia strains and their hosts. In order to identify genes with a potential link to PI, we assembled the genome of an additional PI strain, wLcla. Comparisons between four PI Wolbachia, including wFol and wLcla, and fourteen non-PI Wolbachia yielded a small set of potential candidate genes for further investigation.

CONCLUSIONS: The strong similarities in genome content of wFol and its host, as well as the correlation between host and Wolbachia gene numbers suggest that there may be some form of convergent evolution between endosymbiont and host genomes. If such convergent evolution would be strong enough to overcome the evolutionary forces causing genome reduction, it would enable expanded genomes within long-term obligate endosymbionts.}, } @article {pmid30723148, year = {2019}, author = {Buchman, A and Gamez, S and Li, M and Antoshechkin, I and Li, HH and Wang, HW and Chen, CH and Klein, MJ and Duchemin, JB and Paradkar, PN and Akbari, OS}, title = {Engineered resistance to Zika virus in transgenic Aedes aegypti expressing a polycistronic cluster of synthetic small RNAs.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {116}, number = {9}, pages = {3656-3661}, pmid = {30723148}, issn = {1091-6490}, support = {K22 AI113060/AI/NIAID NIH HHS/United States ; R21 AI123937/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified/genetics/virology ; Disease Outbreaks ; Humans ; Mosquito Vectors/*genetics/virology ; Saliva/virology ; Viral Load/genetics ; Wolbachia/pathogenicity/virology ; Zika Virus/*genetics/pathogenicity ; Zika Virus Infection/*genetics/transmission/virology ; }, abstract = {Recent Zika virus (ZIKV) outbreaks have highlighted the necessity for development of novel vector control strategies to combat arboviral transmission, including genetic versions of the sterile insect technique, artificial infection with Wolbachia to reduce population size and/or vectoring competency, and gene drive-based methods. Here, we describe the development of mosquitoes synthetically engineered to impede vector competence to ZIKV. We demonstrate that a polycistronic cluster of engineered synthetic small RNAs targeting ZIKV is expressed and fully processed in Aedes aegypti, ensuring the formation of mature synthetic small RNAs in the midgut where ZIKV resides in the early stages of infection. Critically, we demonstrate that engineered Ae. aegypti mosquitoes harboring the anti-ZIKV transgene have significantly reduced viral infection, dissemination, and transmission rates of ZIKV. Taken together, these compelling results provide a promising path forward for development of effective genetic-based ZIKV control strategies, which could potentially be extended to curtail other arboviruses.}, } @article {pmid30716462, year = {2019}, author = {Odeniran, PO and Macleod, ET and Ademola, IO and Welburn, SC}, title = {Endosymbionts interaction with trypanosomes in Palpalis group of Glossina captured in southwest Nigeria.}, journal = {Parasitology international}, volume = {70}, number = {}, pages = {64-69}, doi = {10.1016/j.parint.2019.01.011}, pmid = {30716462}, issn = {1873-0329}, mesh = {Animals ; Enterobacteriaceae/*isolation & purification/physiology ; Insect Vectors/microbiology/parasitology ; Nigeria ; Polymerase Chain Reaction ; Prevalence ; *Symbiosis ; Trypanosoma/microbiology/*physiology ; Tsetse Flies/*microbiology/*parasitology ; Wolbachia ; }, abstract = {Glossina species epidemiological studies were conducted in "fly-belt" endemic zone of southwest Nigeria. Two major study areas were identified and four Nzi traps were set in each site for tsetse collection. This study was conducted to determine the prevalence of endosymbionts (Wigglesworthia glossinidia, Sodalis glossinidius and Wolbachia) in natural field-trapped populations of G. p. palpalis and G. tachinoides and investigate the corresponding interactions with African trypanosomes. A total of 64 tsetse flies were collected, these included G. p. palpalis (n = 28) and G. tachinoides (n = 36). Trypanosome infection and endosymbionts of these flies were determined using polymerase chain reaction (PCR) amplification. The infection rates of W. glossinidia was 100.0% in both species, no flies were positive for Wolbachia. Sodalis glossinidius prevalence was similar between the two-tsetse species, with G. p. palpalis and G. tachinoides showing prevalence of 35.7% (95%CI: 20.7-54.2) and 27.8% (95%CI: 15.9-44.0) respectively. No relationship was found between the endosymbionts and trypanosomes in trapped tsetse flies. More studies are needed to enhance the potential control interventions mediated by endosymbionts to reduce parasitic infections.}, } @article {pmid30715337, year = {2019}, author = {Sinha, A and Li, Z and Sun, L and Carlow, CKS}, title = {Complete Genome Sequence of the Wolbachia wAlbB Endosymbiont of Aedes albopictus.}, journal = {Genome biology and evolution}, volume = {11}, number = {3}, pages = {706-720}, pmid = {30715337}, issn = {1759-6653}, mesh = {Aedes/*microbiology ; Animals ; Ankyrins/genetics ; Cell Line ; DNA Transposable Elements ; Genome Size ; *Genome, Bacterial ; Prophages/genetics ; Proteome ; Type IV Secretion Systems ; Wolbachia/*genetics ; }, abstract = {Wolbachia, an alpha-proteobacterium closely related to Rickettsia, is a maternally transmitted, intracellular symbiont of arthropods and nematodes. Aedes albopictus mosquitoes are naturally infected with Wolbachia strains wAlbA and wAlbB. Cell line Aa23 established from Ae. albopictus embryos retains only wAlbB and is a key model to study host-endosymbiont interactions. We have assembled the complete circular genome of wAlbB from the Aa23 cell line using long-read PacBio sequencing at 500× median coverage. The assembled circular chromosome is 1.48 megabases in size, an increase of more than 300 kb over the published draft wAlbB genome. The annotation of the genome identified 1,205 protein coding genes, 34 tRNA, 3 rRNA, 1 tmRNA, and 3 other ncRNA loci. The long reads enabled sequencing over complex repeat regions which are difficult to resolve with short-read sequencing. Thirteen percent of the genome comprised insertion sequence elements distributed throughout the genome, some of which cause pseudogenization. Prophage WO genes encoding some essential components of phage particle assembly are missing, while the remainder are found in five prophage regions/WO-like islands or scattered around the genome. Orthology analysis identified a core proteome of 535 orthogroups across all completed Wolbachia genomes. The majority of proteins could be annotated using Pfam and eggNOG analyses, including ankyrins and components of the Type IV secretion system. KEGG analysis revealed the absence of five genes in wAlbB which are present in other Wolbachia. The availability of a complete circular chromosome from wAlbB will enable further biochemical, molecular, and genetic analyses on this strain and related Wolbachia.}, } @article {pmid30714238, year = {2019}, author = {Fromont, C and Adair, KL and Douglas, AE}, title = {Correlation and causation between the microbiome, Wolbachia and host functional traits in natural populations of drosophilid flies.}, journal = {Molecular ecology}, volume = {28}, number = {7}, pages = {1826-1841}, doi = {10.1111/mec.15041}, pmid = {30714238}, issn = {1365-294X}, support = {BIO 1241099//National Science Foundation/International ; }, mesh = {Animals ; Bacteria/*classification ; Drosophila/classification/*microbiology ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Spiroplasma/genetics ; Symbiosis ; Sympatry ; Transcriptome ; Wolbachia/*genetics ; }, abstract = {Resident microorganisms are known to influence the fitness and traits of animals under controlled laboratory conditions, but the relevance of these findings to wild animals is uncertain. This study investigated the host functional correlates of microbiota composition in a wild community of three sympatric species of mycophagous drosophilid flies, Drosophila falleni, Drosophila neotestacea and Drosophila putrida. Specifically, we quantified bacterial communities and host transcriptomes by parallel 16S rRNA gene amplicon sequencing and RNA-Seq of individual flies. Among-fly variation in microbiota composition did not partition strongly by sex or species, and included multiple modules, that is, sets of bacterial taxa whose abundance varied in concert across different flies. The abundance of bacteria in several modules varied significantly with multiple host transcripts, especially in females, but the identity of the correlated host transcriptional functions differed with host species, including epithelial barrier function in D. falleni, muscle function in D. putrida, and insect growth and development in D. neotestacea. In D. neotestacea, which harbours the endosymbionts Wolbachia and Spiroplasma, Wolbachia promotes the abundance of Spiroplasma, and is positively correlated with abundance of Lactobacillales and Bacteroidales. Furthermore, most correlations between host gene expression and relative abundance of bacterial modules were co-correlated with abundance of Wolbachia (but not Spiroplasma), indicative of an interdependence between host functional traits, microbiota composition and Wolbachia abundance in this species. These data suggest that, in these natural populations of drosophilid flies, different host species interact with microbial communities in functionally different ways that can vary with the abundance of endosymbionts.}, } @article {pmid30708172, year = {2019}, author = {Shaikevich, E and Bogacheva, A and Rakova, V and Ganushkina, L and Ilinsky, Y}, title = {Wolbachia symbionts in mosquitoes: Intra- and intersupergroup recombinations, horizontal transmission and evolution.}, journal = {Molecular phylogenetics and evolution}, volume = {134}, number = {}, pages = {24-34}, doi = {10.1016/j.ympev.2019.01.020}, pmid = {30708172}, issn = {1095-9513}, mesh = {Aedes/microbiology ; Alleles ; Animals ; Anopheles/genetics ; *Biological Evolution ; Culicidae/*microbiology ; Databases, Genetic ; *Gene Transfer, Horizontal ; Haplotypes/genetics ; Humans ; Mosquito Vectors/microbiology ; Multilocus Sequence Typing ; Phylogeny ; *Recombination, Genetic ; Species Specificity ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Many mosquitoes harbour Wolbachia symbionts that could affect the biology of their host in different ways. Evolutionary relationships of mosquitoes' Wolbachia infection, geographical distribution and symbiont prevalence in many mosquito species are not yet clear. Here, we present the results of Wolbachia screening of 17 mosquito species of four genera-Aedes, Anopheles, Coquillettidia and Culex collected from five regions of Eastern Europe and the Caucasus in 2012-2016. Based on multilocus sequence typing (MLST) data previously published and generated in this study, we try to reveal genetic links between mosquitoes' and other hosts' Wolbachia. The Wolbachia symbionts are found in Culex pipiens, Aedes albopictus and Coquillettidia richiardii and for the first time in Aedes cinereus and Aedes cantans, which are important vectors of human pathogens. Phylogenetic analysis demonstrated multiple origins of infection in mosquitoes although the one-allele-criterion approach revealed links among B-supergroup mosquito Wolbachia with allele content of lepidopteran hosts. The MLST gene content of strain wAlbA from the A-supergroup is linked with different ant species. Several cases of intersupergroup recombinations were found. One of them occurred in the wAlbaB strain of Aedes albopictus, which contains the coxA allele of the A-supergroup, whereas other loci, including wsp, belong to supergroup B. Other cases are revealed for non-mosquito symbionts and they exemplified genetic exchanges of A, B and F supergroups. We conclude that modern Wolbachia diversity in mosquitoes and in many other insect taxa is a recent product of strain recombination and symbiont transfers.}, } @article {pmid30708135, year = {2019}, author = {Konecka, E and Olszanowski, Z}, title = {Phylogenetic analysis based on the 16S rDNA, gltA, gatB, and hcpA gene sequences of Wolbachia from the novel host Ceratozetes thienemanni (Acari: Oribatida).}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {70}, number = {}, pages = {175-181}, doi = {10.1016/j.meegid.2019.01.032}, pmid = {30708135}, issn = {1567-7257}, mesh = {Animals ; Bacterial Proteins/genetics ; DNA, Ribosomal/genetics ; Host Microbial Interactions ; Mites/microbiology ; Phylogeny ; Symbiosis/*genetics ; Wolbachia/*classification/genetics ; }, abstract = {We determined the occurrence of intracellular endosymbionts (Wolbachia, Cardinium, Arsenophonus, Rickettsia, Spiroplasma, Hamiltonella, flavobacteria, and microsporidia) in oribatid mites (Acari: Oribatida) with the use of PCR technique. For the first time we looked for and detected Wolbachia in parthenogenetic oribatid mite Ceratozetes thienemanni Willmann, 1943. The 16S rDNA, gatB, hcpA, and gltA sequences of Wolbachia in C. thienemanni showed the highest similarity (≥ 90%) to the genes of Wolbachia from springtails (Collembola) and oribatid mite Gustavia microcephala. We found the unique sequence 5'-GGGGTAATGGCC-3' in 16S rDNA of Wolbachia from C. thienemanni and collembolan representing group E. The phylogeny of Wolbachia based on the analysis of single genes as well as concatenated alignments of four bacterial loci showed that the bacteria from C. thienemanni belonged to Wolbachia group E, like the endosymbionts from springtail hosts and G. microcephala. Considering coexisting of representatives of Oribatida and Collembola in the same soil habitat and similar food, it is possible that the source of Wolbachia infection was the same. Residues of dead invertebrates could be in organic matter of their soil food, so the scenario of infection transferred by eating of remains of soil cohabitates is also possible. It could explain the similarity and relationship of the Wolbachia in these two arthropod groups. Oribatid mite C. thienemanni is a parthenogenetic mite which is a unique feature in the genus Ceratozetes. Moreover, this species, within the entire genus Ceratozetes, is characterized by the most northerly distribution. It is difficult to determine either it is parthenogenesis or the presence of endosymbionts that are in some way responsible for this kind of evolutionary success. Maybe we are dealing here with a kind of synergy of both factors?}, } @article {pmid30700657, year = {2019}, author = {Fukui, Y and Inokuma, H}, title = {Subclinical Infections of Anaplasma phagocytophilum and Anaplasma bovis in Dogs from Ibaraki, Japan.}, journal = {Japanese journal of infectious diseases}, volume = {72}, number = {3}, pages = {168-172}, doi = {10.7883/yoken.JJID.2018.470}, pmid = {30700657}, issn = {1884-2836}, mesh = {Anaplasma/*genetics/isolation & purification ; Anaplasma phagocytophilum/isolation & purification ; Anaplasmosis/blood/*epidemiology ; Animals ; Antibodies, Bacterial/isolation & purification ; DNA, Bacterial/genetics ; Dog Diseases/blood/*epidemiology/*microbiology ; Dogs ; Female ; Fluorescent Antibody Technique/veterinary ; Japan/epidemiology ; Male ; Polymerase Chain Reaction/veterinary ; Prevalence ; }, abstract = {We evaluated the prevalence of Anaplasma infection in 332 dogs from Ibaraki, Japan, using serological and molecular methods. An immunofluorescence antibody assay against Anaplasma phagocytophilum indicated that 7 of the 328 serum samples tested (2.1%) were positive for A. phagocytophilum. Screening by polymerase chain reaction (PCR) analysis demonstrated that 8 of the 331 peripheral blood samples tested (2.4%) were positive for Anaplasmataceae. Phylogenetic analysis of the partial 16S rRNA sequence of the PCR amplicons revealed that 6 sequences were most similar to the 16S rRNA sequence of a Wolbachia sp., and the remaining 2 to A. bovis. Further analysis by A. phagocytophilum-specific nested PCR demonstrated that 1 dog infected with A. bovis was also positive for A. phagocytophilum. This is the first study to report the dual infection of a dog in Japan with A. bovis and A. phagocytophilum.}, } @article {pmid30699194, year = {2019}, author = {Russell, SL and Lemseffer, N and Sullivan, WT}, title = {Correction: Wolbachia and host germline components compete for kinesin-mediated transport to the posterior pole of the Drosophila oocyte.}, journal = {PLoS pathogens}, volume = {15}, number = {1}, pages = {e1007557}, pmid = {30699194}, issn = {1553-7374}, abstract = {[This corrects the article DOI: 10.1371/journal.ppat.1007216.].}, } @article {pmid30696916, year = {2019}, author = {Kajtoch, Ł and Kolasa, M and Kubisz, D and Gutowski, JM and Ścibior, R and Mazur, MA and Holecová, M}, title = {Using host species traits to understand the Wolbachia infection distribution across terrestrial beetles.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {847}, pmid = {30696916}, issn = {2045-2322}, mesh = {Animals ; Bacterial Infections/genetics/*immunology ; Biological Evolution ; Coleoptera/*immunology ; Ecosystem ; Host Specificity ; Host-Pathogen Interactions ; Reproduction ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Knowledge of Wolbachia prevalence with respect to its hosts is restricted mainly to taxonomic/phylogenetic context. In contrast, relations between infection and most host's ecological and biological traits are poorly understood. This study aimed to elaborate on relations between bacteria and its beetle hosts in taxonomic and the ecological contexts. In particular, the goal is to verify which ecological and biological traits of beetles could cause them to be prone to be infected. Verification of Wolbachia infection status across 297 beetle taxa showed that approximately 27% of taxa are infected by supergroups A and B. Only minor support for coevolution between bacteria and its beetle hosts was observed in some genera of beetles, but in general coevolution between beetles and Wolbachia was rejected. Some traits of beetles were found to be unrelated to Wolbachia prevalence (type of range and thermal preferences); some traits were related with ambiguous effects (habitats, distribution, mobility and body size); some were substantially related (reproduction mode and trophy). The aforementioned summary does not show obvious patterns of Wolbachia prevalence and diversity in relation to host taxonomy, biology, and ecology. As both Wolbachia and Coleoptera are diverse groups, this lack of clear patterns is probably a reflection of nature, which is characterised by highly diversified and probably unstable relations.}, } @article {pmid30685209, year = {2019}, author = {Beckmann, JF and Bonneau, M and Chen, H and Hochstrasser, M and Poinsot, D and Merçot, H and Weill, M and Sicard, M and Charlat, S}, title = {The Toxin-Antidote Model of Cytoplasmic Incompatibility: Genetics and Evolutionary Implications.}, journal = {Trends in genetics : TIG}, volume = {35}, number = {3}, pages = {175-185}, pmid = {30685209}, issn = {0168-9525}, support = {R01 GM046904/GM/NIGMS NIH HHS/United States ; R01 GM053756/GM/NIGMS NIH HHS/United States ; R37 GM046904/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Antidotes/chemistry/therapeutic use ; Arthropods/genetics/microbiology ; Bacterial Toxins/chemistry/*genetics ; Culicidae/genetics/microbiology ; Cytoplasm/*genetics/microbiology ; Drosophila/genetics/microbiology ; Gene Transfer Techniques ; Symbiosis/genetics ; Vector Borne Diseases/*genetics/microbiology ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia bacteria inhabit the cells of about half of all arthropod species, an unparalleled success stemming in large part from selfish invasive strategies. Cytoplasmic incompatibility (CI), whereby the symbiont makes itself essential to embryo viability, is the most common of these and constitutes a promising weapon against vector-borne diseases. After decades of theoretical and experimental struggle, major recent advances have been made toward a molecular understanding of this phenomenon. As pieces of the puzzle come together, from yeast and Drosophila fly transgenesis to CI diversity patterns in natural mosquito populations, it becomes clearer than ever that the CI induction and rescue stem from a toxin-antidote (TA) system. Further, the tight association of the CI genes with prophages provides clues to the possible evolutionary origin of this phenomenon and the levels of selection at play.}, } @article {pmid30680117, year = {2019}, author = {Duffy, E and Archer, CR and Sharma, MD and Prus, M and Joag, RA and Radwan, J and Wedell, N and Hosken, DJ}, title = {Wolbachia infection can bias estimates of intralocus sexual conflict.}, journal = {Ecology and evolution}, volume = {9}, number = {1}, pages = {328-338}, pmid = {30680117}, issn = {2045-7758}, abstract = {Males and females share most of their genome and develop many of the same traits. However, each sex frequently has different optimal values for these shared traits, creating intralocus sexual conflict. This conflict has been observed in wild and laboratory populations of insects and affects important evolutionary processes such as sexual selection, the maintenance of genetic variation, and possibly even speciation. Given the broad impacts of intralocus conflict, accurately detecting and measuring it is important. A common way to detect intralocus sexual conflict is to calculate the intersexual genetic correlation for fitness, with negative values suggesting conflict. Here, we highlight a potential confounder of this measure-cytoplasmic incompatibility caused by the intracellular parasite Wolbachia. Infection with Wolbachia can generate negative intersexual genetic correlations for fitness in insects, suggestive of intralocus sexual conflict. This is because cytoplasmic incompatibility reduces the fitness of uninfected females mated to infected males, while uninfected males will not suffer reductions in fitness if they mate with infected females and may even be fitter than infected males. This can lead to strong negative intersexual genetic correlations for fitness, mimicking intralocus conflict. We illustrate this issue using simulations and then present Drosophila simulans data that show how reproductive incompatibilities caused by Wolbachia infection can generate signals of intralocus sexual conflict. Given that Wolbachia infection in insect populations is pervasive, but populations usually contain both infected and uninfected individuals providing scope for cytoplasmic incompatibility, this is an important consideration for sexual conflict research but one which, to date, has been largely underappreciated.}, } @article {pmid30679245, year = {2019}, author = {Gruntenko, NE and Karpova, EK and Adonyeva, NV and Andreenkova, OV and Burdina, EV and Ilinsky, YY and Bykov, RA and Menshanov, PN and Rauschenbach, IY}, title = {Drosophila female fertility and juvenile hormone metabolism depends on the type of Wolbachia infection.}, journal = {The Journal of experimental biology}, volume = {222}, number = {Pt 4}, pages = {}, doi = {10.1242/jeb.195347}, pmid = {30679245}, issn = {1477-9145}, mesh = {Animals ; Drosophila melanogaster/microbiology/*physiology ; Female ; Fertility ; Genotype ; Juvenile Hormones/*metabolism ; Male ; Wolbachia/genetics/*physiology ; }, abstract = {Maternally inherited intracellular bacteria Wolbachia cause both parasitic and mutualistic effects on their numerous insect hosts, including manipulating the host reproductive system in order to increase the bacteria spreading in a host population, and increasing the host fitness. Here, we demonstrate that the type of Wolbachia infection determines the effect on Drosophila melanogaster egg production as a proxy for fecundity, and metabolism of juvenile hormone (JH), which acts as gonadotropin in adult insects. For this study, we used six D. melanogaster lineages carrying the nuclear background of interbred Bi90 lineage and cytoplasmic backgrounds with or without Wolbachia of different genotype variants. The wMelCS genotype of Wolbachia decreases egg production in infected D. melanogaster females in the beginning of oviposition and increases it later (from the sixth day after eclosion), whereas the wMelPop Wolbachia strain causes the opposite effect, and the wMel, wMel2 and wMel4 genotypes of Wolbachia do not show any effect on these traits compared with uninfected Bi90 D. melanogaster females. The intensity of JH catabolism negatively correlates with the fecundity level in the flies carrying both wMelCS and wMelPop Wolbachia The JH catabolism in females infected with genotypes of the wMel group does not differ from that in uninfected females. The effects of wMelCS and wMelPop infection on egg production can be levelled by the modulation of JH titre (via precocene/JH treatment of the flies). Thus, at least one of the mechanisms promoting the effect of Wolbachia on D. melanogaster female fecundity is mediated by JH.}, } @article {pmid30673989, year = {2019}, author = {Fallon, AM}, title = {Conditions facilitating infection of mosquito cell lines with Wolbachia, an obligate intracellular bacterium.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {55}, number = {2}, pages = {120-129}, pmid = {30673989}, issn = {1543-706X}, support = {AI081322//National Institutes of Health/ ; }, mesh = {Aedes/cytology/drug effects/*microbiology ; Aneuploidy ; Animals ; Bacterial Infections/*microbiology ; Cell Line ; Intracellular Space/*microbiology ; Methotrexate/pharmacology ; Wolbachia/drug effects/pathogenicity/*physiology ; }, abstract = {Factors that influence establishment of Wolbachia, an obligate intracellular bacterium, in novel insect hosts or uninfected insect cell lines are poorly understood. Infectivity of Wolbachia strain wStr was correlated with flow cytometric profiles to define optimal conditions for harvesting an infectious inoculum. Wolbachia recovered from the cell culture supernatant after gentle pipetting of infected cells represented about 1% of the total bacterial population and were more infectious than Wolbachia that remained associated with intact cells and/or membranes after low-speed centrifugation. Optimal establishment of a robust infection in naïve cells required 6 d, at a ratio of 80 to 160 bacteria per cell. Among Aedes albopictus mosquito cell lines, an aneuploid line with a 4n + 1 karyotype was more susceptible to infection than diploid lines. These findings contribute to the in vitro manipulation of Wolbachia, illustrate some of the many factors that influence infectivity, and identify areas for future investigation.}, } @article {pmid30668787, year = {2019}, author = {Chebbi, MA and Becking, T and Moumen, B and Giraud, I and Gilbert, C and Peccoud, J and Cordaux, R}, title = {The Genome of Armadillidium vulgare (Crustacea, Isopoda) Provides Insights into Sex Chromosome Evolution in the Context of Cytoplasmic Sex Determination.}, journal = {Molecular biology and evolution}, volume = {36}, number = {4}, pages = {727-741}, doi = {10.1093/molbev/msz010}, pmid = {30668787}, issn = {1537-1719}, mesh = {Animals ; *Biological Evolution ; Female ; *Genome ; Isopoda/*genetics ; Male ; *Sex Chromosomes ; *Sex Determination Processes ; Wolbachia/genetics ; }, abstract = {The terrestrial isopod Armadillidium vulgare is an original model to study the evolution of sex determination and symbiosis in animals. Its sex can be determined by ZW sex chromosomes, or by feminizing Wolbachia bacterial endosymbionts. Here, we report the sequence and analysis of the ZW female genome of A. vulgare. A distinguishing feature of the 1.72 gigabase assembly is the abundance of repeats (68% of the genome). We show that the Z and W sex chromosomes are essentially undifferentiated at the molecular level and the W-specific region is extremely small (at most several hundreds of kilobases). Our results suggest that recombination suppression has not spread very far from the sex-determining locus, if at all. This is consistent with A. vulgare possessing evolutionarily young sex chromosomes. We characterized multiple Wolbachia nuclear inserts in the A. vulgare genome, none of which is associated with the W-specific region. We also identified several candidate genes that may be involved in the sex determination or sexual differentiation pathways. The A. vulgare genome serves as a resource for studying the biology and evolution of crustaceans, one of the most speciose and emblematic metazoan groups.}, } @article {pmid30668658, year = {2019}, author = {Moreira, M and Aguiar, AMF and Bourtzis, K and Latorre, A and Khadem, M}, title = {Wolbachia (Alphaproteobacteria: Rickettsiales) Infections in Isolated Aphid Populations from Oceanic Islands of the Azores Archipelago: Revisiting the Supergroups M and N.}, journal = {Environmental entomology}, volume = {48}, number = {2}, pages = {326-334}, doi = {10.1093/ee/nvy189}, pmid = {30668658}, issn = {1938-2936}, mesh = {Animals ; Aphids/genetics/*microbiology ; Azores ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Aphids (Hemiptera: Aphididae) have provided a suitable model to study endosymbionts, their community, and dynamics since the discovery of the obligate endosymbiont Buchnera aphidicola in these organisms. In previous studies, Wolbachia was found in some aphid species. In the present study, we report the prevalence of Wolbachia in aphids sampled from a geographically isolated region (Azores Islands), aiming at a better understanding and characterization of the two newly reported supergroups, M and N. The description of the supergroup M was based on 16S rRNA as well as some protein-coding genes. However, the assignment of the supergroup N was according to 16S rRNA gene sequences of a very few samples. We collected aphid samples and performed phylogenetic analysis of 16S rRNA gene as well as four protein-coding genes (gatB, ftsZ, coxA, and hcpA). The results demonstrate that the 16S rRNA gene data can unambiguously assign the strain supergroup and that the two supergroups, N and M, are equally prevalent in Azorean aphids. The available sequence data for the protein-coding markers can identify supergroup M but the status of supergroup N is inconclusive, requiring further studies. The data suggest that horizontal transmission of Wolbachia (Hertig and Wolbach) between two phylogenetically distant aphid species cohabiting the same plant host.}, } @article {pmid30650347, year = {2019}, author = {Haqshenas, G and Terradas, G and Paradkar, PN and Duchemin, JB and McGraw, EA and Doerig, C}, title = {A Role for the Insulin Receptor in the Suppression of Dengue Virus and Zika Virus in Wolbachia-Infected Mosquito Cells.}, journal = {Cell reports}, volume = {26}, number = {3}, pages = {529-535.e3}, doi = {10.1016/j.celrep.2018.12.068}, pmid = {30650347}, issn = {2211-1247}, mesh = {Animals ; Culicidae ; Dengue Virus/*pathogenicity ; Receptor, Insulin/pharmacology/*therapeutic use ; Wolbachia/*chemistry ; Zika Virus/*pathogenicity ; }, abstract = {Wolbachia-infected mosquitoes are refractory to super-infection with arthropod-borne pathogens, but the role of host cell signaling proteins in pathogen-blocking mechanisms remains to be elucidated. Here, we use an antibody microarray approach to provide a comprehensive picture of the signaling response of Aedes aegypti-derived cells to Wolbachia. This approach identifies the host cell insulin receptor as being downregulated by the bacterium. Furthermore, siRNA-mediated knockdown and treatment with a small-molecule inhibitor of the insulin receptor kinase concur to assign a crucial role for this enzyme in the replication of dengue and Zika viruses in cultured mosquito cells. Finally, we show that the production of Zika virus in Wolbachia-free live mosquitoes is impaired by treatment with the selective inhibitor mimicking Wolbachia infection. This study identifies Wolbachia-mediated downregulation of insulin receptor kinase activity as a mechanism contributing to the blocking of super-infection by arboviruses.}, } @article {pmid30646951, year = {2019}, author = {da Silva Gonçalves, D and Iturbe-Ormaetxe, I and Martins-da-Silva, A and Telleria, EL and Rocha, MN and Traub-Csekö, YM and O'Neill, SL and Sant'Anna, MRV and Moreira, LA}, title = {Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {33}, pmid = {30646951}, issn = {1756-3305}, mesh = {Animals ; Cell Line ; *Gene Expression ; *Immunity, Innate ; Immunologic Factors/*biosynthesis ; Leishmania infantum/*growth & development ; *Microbial Interactions ; Parasite Load ; Psychodidae/*immunology/microbiology ; Wolbachia/growth & development/*immunology ; }, abstract = {BACKGROUND: The leishmaniases are important neglected diseases caused by Leishmania spp. which are transmitted by sand flies, Lutzomyia longipalpis being the main vector of visceral leishmaniasis in the Americas. The methodologies for leishmaniasis control are not efficient, causing 1.5 million reported cases annually worldwide, therefore showing the need for development of novel strategies and interventions to control transmission of the disease. The bacterium Wolbachia pipientis is being used to control viruses transmitted by mosquitoes, such as dengue and Zika, and its introduction in disease vectors has been effective against parasites such as Plasmodium. Here we show the first successful establishment of Wolbachia into two different embryonic cell lines from L. longipalpis, LL-5 and Lulo, and analysed its effects on the sand fly innate immune system, followed by in vitro Leishmania infantum interaction.

RESULTS: Our results show that LL-5 cells respond to wMel and wMelPop-CLA strains within the first 72 h post-infection, through the expression of antimicrobial peptides and inducible nitric oxide synthase resulting in a decrease of Wolbachia detection in the early stages of infection. In subsequent passages, the wMel strain was not able to infect any of the sand fly cell lines while the wMelPop-CLA strain was able to stably infect Lulo cells and LL-5 at lower levels. In Wolbachia stably infected cells, the expression of immune-related genes involved with downregulation of the IMD, Toll and Jak-Stat innate immune pathways was significantly decreased, in comparison with the uninfected control, suggesting immune activation upon Wolbachia transinfection. Furthermore, Wolbachia transinfection did not promote a negative effect on parasite load in those cells.

CONCLUSIONS: Initial strong immune responses of LL5 cells might explain the inefficiency of stable infections in these cells while we found that Lulo cells are more permissive to infection with Wolbachia causing an effect on the cell immune system, but not against in vitro L. infantum interaction. This establishes Lulo cells as a good system for the adaptation of Wolbachia in L. longipalpis.}, } @article {pmid30644356, year = {2019}, author = {Shaikevich, E and Bogacheva, A and Ganushkina, L}, title = {Dirofilaria and Wolbachia in mosquitoes (Diptera: Culicidae) in central European Russia and on the Black Sea coast.}, journal = {Parasite (Paris, France)}, volume = {26}, number = {}, pages = {2}, pmid = {30644356}, issn = {1776-1042}, mesh = {Animals ; Black Sea ; Culex/microbiology/parasitology ; Culicidae/*microbiology/*parasitology ; DNA, Helminth/isolation & purification ; Dirofilaria immitis/genetics/isolation & purification/physiology ; Dirofilaria repens/isolation & purification/physiology ; Dirofilariasis/*epidemiology/*transmission ; Dog Diseases/epidemiology ; Dogs ; Female ; Larva/microbiology/parasitology ; Mosquito Vectors/microbiology/parasitology ; Russia/epidemiology ; Symbiosis ; Wolbachia/genetics/*isolation & purification/physiology ; }, abstract = {Dirofilariasis is endemic in Russia, as well as in many other European countries. The aim of this study was to assess the ability of mosquitoes to transfer Dirofilaria immitis and Dirofilaria repens in regions with temperate and subtropical climates. The possible impact of the symbiotic bacterium Wolbachia on Dirofilaria transmission was also investigated. 5333 female mosquitoes were collected at 11 points in central European Russia and on the Black Sea coast during the period 2013-2017. Out of 20 mosquito species examined, 14 were infected with D. repens and 13 with D. immitis. Both species of Dirofilaria were found in different climatic regions. The total Dirofilaria spp. estimated infection rate (EIR) in the central part of Russia varied from 3.1% to 3.7% and, in the southern region, from 1.1% to 3.0%. The highest estimated infection rate was found in Anopheles messeae, the lowest in Culex pipiens. The greatest epidemiological danger was represented by Aedes aegypti, Ae. geniculatus, An. messeae and Ae. communis. Six out of 20 mosquito species were infected with Wolbachia. Pools of Aedes albopictus, Cx. pipiens and Coquillettidia richiardii were simultaneously infected with Dirofilaria and Wolbachia. After checking mosquitoes individually, it was found that there was no development of Dirofilaria to the infective larval stage in specimens infected with Wolbachia. Twenty-two Dirofilaria-infective pools were Wolbachia-free and only two mosquito pools were Wolbachia-infected. The potential for transmission of Dirofilaria in mosquito species naturally uninfected with the symbiotic bacterium Wolbachia is higher than in species infected with the bacterium.}, } @article {pmid30642130, year = {2019}, author = {Foo, IJ and Hoffmann, AA and Ross, PA}, title = {Cross-Generational Effects of Heat Stress on Fitness and Wolbachia Density in Aedes aegypti Mosquitoes.}, journal = {Tropical medicine and infectious disease}, volume = {4}, number = {1}, pages = {}, pmid = {30642130}, issn = {2414-6366}, support = {1132412//National Health and Medical Research Council/ ; 1118640//National Health and Medical Research Council/ ; LE150100083//Australian Research Council/ ; }, abstract = {Aedes aegypti mosquitoes infected with Wolbachia symbionts are now being released into the field to control the spread of pathogenic human arboviruses. Wolbachia can spread throughout vector populations by inducing cytoplasmic incompatibility and can reduce disease transmission by interfering with virus replication. The success of this strategy depends on the effects of Wolbachia on mosquito fitness and the stability of Wolbachia infections across generations. Wolbachia infections are vulnerable to heat stress, and sustained periods of hot weather in the field may influence their utility as disease control agents, particularly if temperature effects persist across generations. To investigate the cross-generational effects of heat stress on Wolbachia density and mosquito fitness, we subjected Ae. aegypti with two different Wolbachia infection types (wMel, wAlbB) and uninfected controls to cyclical heat stress during larval development over two generations. We then tested adult starvation tolerance and wing length as measures of fitness and measured the density of wMel in adults. Both heat stress and Wolbachia infection reduced adult starvation tolerance. wMel Wolbachia density in female offspring was lower when mothers experienced heat stress, but male Wolbachia density did not depend on the rearing temperature of the previous generation. We also found cross-generational effects of heat stress on female starvation tolerance, but there was no cross-generational effect on wing length. Fitness costs of Wolbachia infections and cross-generational effects of heat stress on Wolbachia density may reduce the ability of Wolbachia to invade populations and control arbovirus transmission under specific environmental conditions.}, } @article {pmid30632016, year = {2019}, author = {Öhlund, P and Lundén, H and Blomström, AL}, title = {Insect-specific virus evolution and potential effects on vector competence.}, journal = {Virus genes}, volume = {55}, number = {2}, pages = {127-137}, pmid = {30632016}, issn = {1572-994X}, support = {2016-01251//Vetenskapsrådet/ ; vice chancellor career support//Sveriges Lantbruksuniversitet/ ; }, mesh = {Animals ; Arboviruses/*genetics/pathogenicity ; Culicidae/genetics/virology ; Flaviviridae/genetics/pathogenicity ; High-Throughput Nucleotide Sequencing ; Insect Vectors/virology ; Insect Viruses/*genetics/pathogenicity ; Pest Control, Biological ; Species Specificity ; Togaviridae/genetics/pathogenicity ; Virus Diseases/*genetics/virology ; Virus Replication/*genetics ; }, abstract = {The advancement in high-throughput sequencing technology and bioinformatics tools has spurred a new age of viral discovery. Arthropods is the largest group of animals and has shown to be a major reservoir of different viruses, including a group known as insect-specific viruses (ISVs). The majority of known ISVs have been isolated from mosquitoes and shown to belong to viral families associated with animal arbovirus pathogens, such as Flaviviridae, Togaviridae and Phenuiviridae. These insect-specific viruses have a strict tropism and are unable to replicate in vertebrate cells, these properties are interesting for many reasons. One is that these viruses could potentially be utilised as biocontrol agents using a similar strategy as for Wolbachia. Mosquitoes infected with the viral agent could have inferior vectorial capacity of arboviruses resulting in a decrease of circulating arboviruses of public health importance. Moreover, insect-specific viruses are thought to be ancestral to arboviruses and could be used to study the evolution of the switch from single-host to dual-host. In this review, we discuss new discoveries and hypothesis in the field of arboviruses and insect-specific viruses.}, } @article {pmid30629155, year = {2019}, author = {Sazama, EJ and Ouellette, SP and Wesner, JS}, title = {Bacterial Endosymbionts Are Common Among, but not Necessarily Within, Insect Species.}, journal = {Environmental entomology}, volume = {48}, number = {1}, pages = {127-133}, doi = {10.1093/ee/nvy188}, pmid = {30629155}, issn = {1938-2936}, mesh = {Animals ; Insecta/*microbiology ; Linear Models ; Rickettsiales ; *Symbiosis ; Wolbachia ; }, abstract = {Bacterial endosymbionts, particularly Wolbachia (Rickettsiales: Rickettsiaceae), Rickettsia (Rickettsiales: Rickettsiaceae), and Cardinium (Bacteroidales: Bacteroidaceae), are commonly found in several arthropod groups, including insects. Most estimates of the global infection rate of Wolbachia (52% [95% credible intervals: 44-60]) show that these bacteria infect more than half of all insect species. Other endosymbionts, such as Rickettsia (24% [confidence intervals [CIs] 20-42]) and Cardinium (13% [CIs 13-55]), infect a smaller but still substantial proportion of insect species. In spite of these observations, it is unclear what proportion of individuals within those species are infected. Here, we used published databases to estimate the proportion of individuals that are infected with either Wolbachia, Rickettsia, or Cardinium. We found that the majority (69%) of Wolbachia-infected species have less than half of their individuals infected with Wolbachia, indicating that although the bacterium may be common among species, it is not common within species. The same was true for Rickettsia (81%) and Cardinium (87%). This discrepancy was consistent across orders, in which less than 10% of individuals were typically infected, even though more than 50% of species within orders were infected. For example, according to our model, nearly 50% of beetle (Coleoptera) species are infected with Wolbachia (i.e., contain at least one individual that has tested positive for Wolbachia), but less than 5% of all individuals are infected. These results add to the growing knowledge base about endosymbionts in insects and should guide future sampling efforts and investigations on the role that these bacteria play in populations.}, } @article {pmid30624116, year = {2018}, author = {Dossi, FCA and da Silva, EP and Cônsoli, FL}, title = {Shifting the Balance: Heat Stress Challenges the Symbiotic Interactions of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera, Liviidae).}, journal = {The Biological bulletin}, volume = {235}, number = {3}, pages = {195-203}, doi = {10.1086/699755}, pmid = {30624116}, issn = {1939-8697}, mesh = {Animals ; Hemiptera/*physiology ; *Hot Temperature ; Symbiosis/*physiology ; }, abstract = {Global warming may impact biodiversity by disrupting biological interactions, including long-term insect-microbe mutualistic associations. Symbiont-mediated insect tolerance to high temperatures is an ecologically important trait that significantly influences an insect's life history. Disruption of microbial symbionts that are required by insects would substantially impact their pest status. Diaphorina citri, a worldwide citrus pest, is associated with the mutualistic symbionts Candidatus Carsonella ruddii and Candidatus Profftella armatura. Wolbachia is also associated with D. citri, but its contribution to the host is unknown. Symbiont density is dependent on a range of factors, including the thermosensitivity of the host and/or symbiont to heat stress. Here, we predicted that short-term heat stress of D. citri would disrupt the host-symbiont phenological synchrony and differentially affect the growth and density of symbionts. We investigated the effects of exposing D. citri eggs to different temperatures for different periods of time on the growth dynamics of symbionts during the nymphal development of D. citri (first instar to fifth instar) by real-time polymerase chain reaction. Symbiont densities were assessed as the number of gene copies, using specific molecular markers: 16S rRNA for Carsonella and Profftella and ftsZ for Wolbachia. Statistical modeling of the copy numbers of symbionts revealed differences in their growth patterns, particularly in the early instars of heat-shocked insects. Wolbachia was the only symbiont to benefit from heat-shock treatment. Although the symbionts responded differently to heat stress, the lack of differences in symbiont densities between treated and control late nymphs suggests the existence of an adaptive genetic process to restore phenological synchrony during the development of immatures in preparation for adult life. Our findings contribute to the understanding of the potential deleterious effects of high temperatures on host-symbiont interactions. Our data also suggest that the effects of host exposure to high temperatures in symbiont growth are highly variable and dependent on the interactions among members of the community of symbionts harbored by a host. Such dependence points to unpredictable consequences for agroecosystems worldwide due to climate change-related effects on the ecological traits of symbiont-dependent insect pests.}, } @article {pmid30622893, year = {2019}, author = {Millán, J and Travaini, A and Cevidanes, A and Sacristán, I and Rodríguez, A}, title = {Assessing the natural circulation of canine vector-borne pathogens in foxes, ticks and fleas in protected areas of Argentine Patagonia with negligible dog participation.}, journal = {International journal for parasitology. Parasites and wildlife}, volume = {8}, number = {}, pages = {63-70}, pmid = {30622893}, issn = {2213-2244}, abstract = {We collected blood and/or ectoparasites from 49 South American grey foxes (Lycalopex griseus) and two Andean foxes (L. culpaeus) caught in two National Parks of southern Argentine Patagonia (Bosques Petrificados, BPNP; and Monte León, MLNP) where dogs are nearly absent (density < 0.01 dog/km[2]). Common ectoparasites were the flea Pulex irritans (88% prevalence) and the tick Amblyomma tigrinum (29%). Conventional PCR and sequencing of 49 blood samples, 299 fleas analysed in 78 pools, and 21 ticks revealed the presence of DNA of the following canine vector-borne pathogens: in grey foxes, Rickettsia sp. (3%), hemoplasmas (8%), including Mycoplasma haemocanis, and Hepatozoon sp. (50%); in P. irritans, Bartonella spp. (72% of flea pools from 76% of foxes), mostly B. vinsonii subsp. berkhoffii but also B. rochalimae, Anaplasmataceae (Wolbachia sp.; 60% and 54%), and M. haemocanis/haemofelis (29% and 18%); and in A. tigrinum, Hepatozoon sp. (33% of ticks in 4 of 7 foxes). No piroplasmid DNA was detected in any sample. Andean foxes were negative for all tested pathogens. Two different Hepatozoon haplotypes were detected: the most prevalent was phylogenetically associated with H. felis, and the other with H. americanum and related sequences. Amblyomma tigrinum and Hepatozoon sp. were more abundant and/or prevalent in BPNP than in colder MLNP, 300 km southwards, perhaps located close to the limit for tick suitability. Bartonella v. berkhoffii was also significantly more prevalent in fleas of foxes in BPNP than in MLNP. This study provides novel information about natural host-pathogen associations in wildlife, markedly extends the distribution area in South America of arthropods and vector-borne pathogens of veterinary and public health interest, and contributes preliminary evidence about the potential role of A. tigrinum and P. irritans as vectors, respectively, for potentially new species of Hepatozoon from Lycalopex spp. and for M. haemocanis that should be further investigated.}, } @article {pmid30622399, year = {2018}, author = {Kirichenko, N and Triberti, P and Kobayashi, S and Hirowatari, T and Doorenweerd, C and Ohshima, I and Huang, GH and Wang, M and Magnoux, E and Lopez-Vaamonde, C}, title = {Systematics of Phyllocnistis leaf-mining moths (Lepidoptera, Gracillariidae) feeding on dogwood (Cornus spp.) in Northeast Asia, with the description of three new species.}, journal = {ZooKeys}, volume = {}, number = {736}, pages = {79-118}, pmid = {30622399}, issn = {1313-2989}, abstract = {During an ongoing DNA-barcoding campaign of the leaf-mining moths that feed on woody plants in Northeast Asia, four lineages of the genus Phyllocnistis (Gracillariidae, Phyllocnistinae) were discovered on dogwood (Cornus spp): P. cornella Ermolaev, 1987 on C. controversa Hemsl. (Japan: Hokkaido) and three new species - one feeding on C. controversa, C. florida L. and C. macrophylla Wall. in Japan (Honshu, Shikoku, Kyushu), a second species on C. macrophylla in China (Yunnan) and a third on Siberian dogwood Cornus alba L. in Russia (Siberia). All these species showed differences in morphology, in the barcode region of the cytochrome c oxidase I gene and in two nuclear genes (histone H3 and 28S ribosomal RNA). No correlation was found between the deep mitochondrial splits observed and the Wolbachia infection pattern. Based on both morphological and molecular evidence, the three recently discovered lineages are described here as new species: P. indistincta Kobayashi & Triberti, sp. n. (Japan), P. saepta Kirichenko, Ohshima & Huang, sp. n. (China) and P. verae Kirichenko, Triberti & Lopez-Vaamonde, sp. n. (Russia). In addition, the authors re-describe the adult morphology of P. cornella, provide the first record of this species from Japan and highlight the diagnostic characters that allow these Cornus-feeding Phyllocnistis species to be distinguished.}, } @article {pmid30620733, year = {2019}, author = {Garcia, GA and Sylvestre, G and Aguiar, R and da Costa, GB and Martins, AJ and Lima, JBP and Petersen, MT and Lourenço-de-Oliveira, R and Shadbolt, MF and Rašić, G and Hoffmann, AA and Villela, DAM and Dias, FBS and Dong, Y and O'Neill, SL and Moreira, LA and Maciel-de-Freitas, R}, title = {Matching the genetics of released and local Aedes aegypti populations is critical to assure Wolbachia invasion.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {1}, pages = {e0007023}, pmid = {30620733}, issn = {1935-2735}, mesh = {Aedes/*drug effects/genetics/*virology ; Animals ; Arboviruses/*growth & development ; Biological Control Agents ; Brazil ; DNA, Mitochondrial/genetics ; Female ; Insecticide Resistance/*genetics ; Male ; Mosquito Vectors/virology ; Pest Control, Biological/*methods ; Pyrethrins/pharmacology ; Wolbachia/*growth & development ; }, abstract = {BACKGROUND: Traditional vector control approaches such as source reduction and insecticide spraying have limited effect on reducing Aedes aegypti population. The endosymbiont Wolbachia is pointed as a promising tool to mitigate arbovirus transmission and has been deployed worldwide. Models predict a rapid increase on the frequency of Wolbachia-positive Ae. aegypti mosquitoes in local settings, supported by cytoplasmic incompatibility (CI) and high maternal transmission rate associated with the wMelBr strain.

Wolbachia wMelBr strain was released for 20 consecutive weeks after receiving >87% approval of householders of the isolated community of Tubiacanga, Rio de Janeiro. wMelBr frequency plateued~40% during weeks 7-19, peaked 65% but dropped as releases stopped. A high (97.56%) maternal transmission was observed. Doubling releases and deploying mosquitoes with large wing length and low laboratory mortality produced no detectable effects on invasion trend. By investigating the lab colony maintenance procedures backwardly, pyrethroid resistant genotypes in wMelBr decreased from 68% to 3.5% after 17 generations. Therefore, we initially released susceptible mosquitoes in a local population highly resistant to pyrethroids which, associated with the over use of insecticides by householders, ended jeopardizing Wolbachia invasion. A new strain (wMelRio) was produced after backcrossing wMelBr females with males from field to introduce mostly pyrethroid resistance alleles. The new strain increased mosquito survival but produced relevant negative effects on Ae. aegypti fecundity traits, reducing egg clutche size and egg hatch. Despite the cost on fitness, wMelRio successful established where wMelBr failed, revealing that matching the local population genetics, especially insecticide resistance background, is critical to achieve invasion.

CONCLUSIONS/SIGNIFICANCE: Local householders support was constantly high, reaching 90% backing on the second release (wMelRio strain). Notwithstanding the drought summer, the harsh temperature recorded (daily average above 30°C) did not seem to affect the expression of maternal transmission of wMel on a Brazilian background. Wolbachia deployment should match the insecticide resistance profile of the wild population to achieve invasion. Considering pyrethroid-resistance is a widely distributed phenotype in natural Ae. aegypti populations, future Wolbachia deployments must pay special attention in maintaining insecticide resistance in lab colonies for releases.}, } @article {pmid30617067, year = {2019}, author = {Hong, WD and Benayoud, F and Nixon, GL and Ford, L and Johnston, KL and Clare, RH and Cassidy, A and Cook, DAN and Siu, A and Shiotani, M and Webborn, PJH and Kavanagh, S and Aljayyoussi, G and Murphy, E and Steven, A and Archer, J and Struever, D and Frohberger, SJ and Ehrens, A and Hübner, MP and Hoerauf, A and Roberts, AP and Hubbard, ATM and Tate, EW and Serwa, RA and Leung, SC and Qie, L and Berry, NG and Gusovsky, F and Hemingway, J and Turner, JD and Taylor, MJ and Ward, SA and O'Neill, PM}, title = {AWZ1066S, a highly specific anti-Wolbachia drug candidate for a short-course treatment of filariasis.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {116}, number = {4}, pages = {1414-1419}, pmid = {30617067}, issn = {1091-6490}, support = {MC_PC_16052/MRC_/Medical Research Council/United Kingdom ; MC_PC_17167/MRC_/Medical Research Council/United Kingdom ; MR/R025401/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Elephantiasis, Filarial/drug therapy/microbiology ; Female ; Male ; Mice ; Mice, SCID ; Onchocerciasis/drug therapy/microbiology ; Pyrimidines/pharmacology ; Quinazolines/pharmacology ; Wolbachia/*drug effects ; }, abstract = {Onchocerciasis and lymphatic filariasis are two neglected tropical diseases that together affect ∼157 million people and inflict severe disability. Both diseases are caused by parasitic filarial nematodes with elimination efforts constrained by the lack of a safe drug that can kill the adult filaria (macrofilaricide). Previous proof-of-concept human trials have demonstrated that depleting >90% of the essential nematode endosymbiont bacterium, Wolbachia, using antibiotics, can lead to permanent sterilization of adult female parasites and a safe macrofilaricidal outcome. AWZ1066S is a highly specific anti-Wolbachia candidate selected through a lead optimization program focused on balancing efficacy, safety and drug metabolism/pharmacokinetic (DMPK) features of a thienopyrimidine/quinazoline scaffold derived from phenotypic screening. AWZ1066S shows superior efficacy to existing anti-Wolbachia therapies in validated preclinical models of infection and has DMPK characteristics that are compatible with a short therapeutic regimen of 7 days or less. This candidate molecule is well-positioned for onward development and has the potential to make a significant impact on communities affected by filariasis.}, } @article {pmid30609847, year = {2019}, author = {Bustamante-Brito, R and Vera-Ponce de León, A and Rosenblueth, M and Martínez-Romero, JC and Martínez-Romero, E}, title = {Metatranscriptomic Analysis of the Bacterial Symbiont Dactylopiibacterium carminicum from the Carmine Cochineal Dactylopius coccus (Hemiptera: Coccoidea: Dactylopiidae).}, journal = {Life (Basel, Switzerland)}, volume = {9}, number = {1}, pages = {}, pmid = {30609847}, issn = {2075-1729}, support = {IN207718//Universidad Nacional Autónoma de México/ ; 253116//Consejo Nacional de Ciencia y Tecnología/ ; }, abstract = {The scale insect Dactylopius coccus produces high amounts of carminic acid, which has historically been used as a pigment by pre-Hispanic American cultures. Nowadays carmine is found in food, cosmetics, and textiles. Metagenomic approaches revealed that Dactylopius spp. cochineals contain two Wolbachia strains, a betaproteobacterium named Candidatus Dactylopiibacterium carminicum and Spiroplasma, in addition to different fungi. We describe here a transcriptomic analysis indicating that Dactylopiibacterium is metabolically active inside the insect host, and estimate that there are over twice as many Dactylopiibacterium cells in the hemolymph than in the gut, with even fewer in the ovary. Albeit scarce, the transcripts in the ovaries support the presence of Dactylopiibacterium in this tissue and a vertical mode of transmission. In the cochineal, Dactylopiibacterium may catabolize plant polysaccharides, and be active in carbon and nitrogen provisioning through its degradative activity and by fixing nitrogen. In most insects, nitrogen-fixing bacteria are found in the gut, but in this study they are shown to occur in the hemolymph, probably delivering essential amino acids and riboflavin to the host from nitrogen substrates derived from nitrogen fixation.}, } @article {pmid30602718, year = {2019}, author = {Clare, RH and Bardelle, C and Harper, P and Hong, WD and Börjesson, U and Johnston, KL and Collier, M and Myhill, L and Cassidy, A and Plant, D and Plant, H and Clark, R and Cook, DAN and Steven, A and Archer, J and McGillan, P and Charoensutthivarakul, S and Bibby, J and Sharma, R and Nixon, GL and Slatko, BE and Cantin, L and Wu, B and Turner, J and Ford, L and Rich, K and Wigglesworth, M and Berry, NG and O'Neill, PM and Taylor, MJ and Ward, SA}, title = {Industrial scale high-throughput screening delivers multiple fast acting macrofilaricides.}, journal = {Nature communications}, volume = {10}, number = {1}, pages = {11}, pmid = {30602718}, issn = {2041-1723}, support = {MC_PC_17167/MRC_/Medical Research Council/United Kingdom ; MR/R025401/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes ; Animals ; Cell Line ; *Drug Discovery ; Filaricides/*analysis ; *High-Throughput Screening Assays ; Wolbachia ; }, abstract = {Nematodes causing lymphatic filariasis and onchocerciasis rely on their bacterial endosymbiont, Wolbachia, for survival and fecundity, making Wolbachia a promising therapeutic target. Here we perform a high-throughput screen of AstraZeneca's 1.3 million in-house compound library and identify 5 novel chemotypes with faster in vitro kill rates (<2 days) than existing anti-Wolbachia drugs that cure onchocerciasis and lymphatic filariasis. This industrial scale anthelmintic neglected tropical disease (NTD) screening campaign is the result of a partnership between the Anti-Wolbachia consortium (A∙WOL) and AstraZeneca. The campaign was informed throughout by rational prioritisation and triage of compounds using cheminformatics to balance chemical diversity and drug like properties reducing the chance of attrition from the outset. Ongoing development of these multiple chemotypes, all with superior time-kill kinetics than registered antibiotics with anti-Wolbachia activity, has the potential to improve upon the current therapeutic options and deliver improved, safer and more selective macrofilaricidal drugs.}, } @article {pmid30596784, year = {2018}, author = {Fortin, M and Vitet, C and Souty-Grosset, C and Richard, FJ}, title = {How do familiarity and relatedness influence mate choice in Armadillidium vulgare?.}, journal = {PloS one}, volume = {13}, number = {12}, pages = {e0209893}, pmid = {30596784}, issn = {1932-6203}, mesh = {Animals ; Female ; Isopoda/microbiology/*physiology ; Male ; Mating Preference, Animal/*physiology ; Wolbachia ; }, abstract = {Mate choice is an important process in sexual selection and usually prevents inbreeding depression in populations. In the terrestrial isopod Armadillidium vulgare, the close physical proximity between individuals may increase the risk of reproducing with siblings. Moreover, individuals of this species can be infected with the feminizing bacteria of Wolbachia, which influence male mate choice. However, little is known about the kinship or familiarity assessment of the selected partner that occurs when a male can choose between females with or without Wolbachia. To investigate the potential mechanisms leading to mate choice and the potential impact of the parasite, we performed behavioral choice tests on males where they could choose between sibling vs. nonsibling females, familiar vs. unfamiliar females, and sibling familiar vs. unfamiliar nonsibling females. To investigate the costs of inbreeding, we compared the reproductive success of both sibling and nonsibling mates. Our results revealed that male copulation attempts were higher for familiar females and for nonsibling females when both females were Wolbachia-infected, but the duration was longer when both females were Wolbachia-free. When males mated with a sibling female, their fecundity was severely decreased, consistent with inbreeding depression. Overall, we observed copulations with all types of females and demonstrated discrimination capacities and potential preferences. We highlight the complexity of the tradeoff between kinship, familiarity and parasite transmission assessment for mate choice.}, } @article {pmid30595344, year = {2019}, author = {Gangwar, M and Jha, R and Goyal, M and Srivastava, M}, title = {Immunogenicity and protective efficacy of Recombinase A from Wolbachia endosymbiont of filarial nematode Brugia malayi (wBmRecA).}, journal = {Vaccine}, volume = {37}, number = {4}, pages = {571-580}, doi = {10.1016/j.vaccine.2018.12.015}, pmid = {30595344}, issn = {1873-2518}, mesh = {Animals ; Antibodies, Helminth/blood ; B-Lymphocytes/immunology ; Brugia malayi/*microbiology ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cloning, Molecular ; Cytokines/immunology ; Elephantiasis, Filarial/immunology/prevention & control ; Female ; *Immunogenicity, Vaccine ; Immunoglobulin G/blood ; Mice ; Rec A Recombinases/genetics/*immunology ; Spleen/immunology ; Wolbachia/*enzymology ; }, abstract = {Lymphatic filariasis causes global morbidity. Wolbachia, an endo-symbiotic intracellular bacterium of the filarial nematode helps in their growth and development, regulates fecundity in female worms and contributes to the immunopathogenesis of the disease. However, genes and proteins of Wolbachia that may act as putative vaccine candidates are not known. In this study, we cloned recombinase-A protein of Wolbachia from Brugia malayi (wBmRecA) and carried out its detailed biochemical and immunological characterization. Bioinformatics analysis, circular dichroism and fluorescence spectral studies showed significant sequence and structural similarities between wBmRecA and RecA of other alpha-proteo- bacterial species. wBmRecA was ubiquitously expressed in all the three major life stages of B. malayi, including excretory-secretory products of the adult worm. In silico studies suggested immunogenic potential of wBmRecA, and mice immunized with wBmRecA exhibited elevated levels of immunoglobulins IgG1, IgG2a, IgG2b and IgG3 in their serum along with increased percentages of CD4[+], CD8[+] T cells and CD19[+] B cells in their spleens. Notably, splenocytes from immunized mice showed increased m-RNA expression of T-bet, elevated proinflammatory cytokines IFN-γ and IL-12, while peritoneal MФs exhibited increased levels of iNOS, downregulated Arg-1 and secreted copious amounts of nitric oxide which contributed to severely impaired development of the infective larvae (Bm-L3). Interestingly, sera from immunized mice promoted significant cellular adherence and cytotoxicity against microfilariae and Bm-L3. Importantly, wBmRecA demonstrated strong immuno-reactivity with bancroftian sera from endemic normal individuals. These results suggest that wBmRecA is highly immunogenic, and should be explored further as a putative vaccine candidate against lymphatic filariasis.}, } @article {pmid30587225, year = {2018}, author = {Savadelis, MD and Day, KM and Bradner, JL and Wolstenholme, AJ and Dzimianski, MT and Moorhead, AR}, title = {Efficacy and side effects of doxycycline versus minocycline in the three-dose melarsomine canine adulticidal heartworm treatment protocol.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {671}, pmid = {30587225}, issn = {1756-3305}, support = {054333-01//American Heartworm Society/ ; }, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage ; Arsenicals/*administration & dosage ; Dirofilaria immitis/*drug effects/microbiology/physiology ; Dirofilariasis/*drug therapy/parasitology ; Dog Diseases/*drug therapy/parasitology ; Dogs ; Doxycycline/*administration & dosage/adverse effects ; Drug Therapy, Combination ; Female ; Filaricides/*administration & dosage ; Male ; Minocycline/*administration & dosage/adverse effects ; Triazines/*administration & dosage ; Wolbachia/*drug effects/genetics/physiology ; }, abstract = {BACKGROUND: The American Heartworm Society currently recommends the use of a monthly macrocyclic lactone, a 28-day course of 10 mg/kg doxycycline BID, and the 3-dose protocol of melarsomine dihydrochloride for the treatment of canine heartworm disease. Doxycycline is necessary for the reduction of the bacterium Wolbachia, found in all heartworm life-stages. Previous price increases and decreasing availability prompted us to evaluate alternative tetracycline antibiotics, i.e. minocycline, for the reduction of Wolbachia during canine heartworm treatment.

METHODS: Thirty-two heartworm-positive dogs were randomized to receive 10 mg/kg or 5 mg/kg of either doxycycline or minocycline for 28 days BID, for a total of 8 dogs per experimental group. All dogs received 6 months of Heartgard Plus® (ivermectin/pyrantel) and the 3-dose protocol of 2.5 mg/kg melarsomine dihydrochloride. Blood samples were collected prior to the initiation of treatment, every 7 days throughout tetracycline treatment, and then monthly thereafter until the dog tested negative for the presence of heartworm antigen. DNA was isolated from circulating microfilarial samples and qPCR was performed on each sample.

RESULTS: A greater number of dogs in the 10 mg/kg doxycycline and minocycline treated groups experienced gastrointestinal side effects as compared to the 5 mg/kg doxycycline and minocycline treated groups. All eight dogs in the 10 mg/kg doxycycline-treated group tested negative for the presence of Wolbachia DNA by 28 days post-tetracycline treatment. A total of two dogs in both the 5 mg/kg doxycycline- and 10 mg/kg minocycline-treated groups and three dogs in the 5 mg/kg minocycline-treated group remained positive for the presence of Wolbachia DNA by the end of tetracycline treatment.

CONCLUSIONS: No lung pathology was assessed in this clinical trial, therefore the clinical effect of the remaining Wolbachia DNA in the 10 mg/kg minocycline-, 5 mg/kg doxycycline- and 5 mg/kg minocycline-treated groups cannot be determined. Owner compliance in the proper administration of these tetracyclines may be impacted by the increased severe gastrointestinal side effects reported for the 10 mg/kg doxycycline- and minocycline-treated groups. We recommend that veterinarians prescribe the recommended 10 mg/kg doxycycline for canine heartworm treatment and reduce the dosage to 5 mg/kg in cases of severe gastrointestinal side effects in order to improve owner compliance in administration of medications.}, } @article {pmid30585387, year = {2018}, author = {Altinli, M and Soms, J and Ravallec, M and Justy, F and Bonneau, M and Weill, M and Gosselin-Grenet, AS and Sicard, M}, title = {Sharing cells with Wolbachia: the transovarian vertical transmission of Culex pipiens densovirus.}, journal = {Environmental microbiology}, volume = {}, number = {}, pages = {}, doi = {10.1111/1462-2920.14511}, pmid = {30585387}, issn = {1462-2920}, support = {//Campus France/ ; //Ministère des affaires étrangères/ ; ANR-10-INBS-04//French National Research Agency/ ; }, abstract = {Culex pipiens densovirus (CpDV), a single stranded DNA virus, has been isolated from Culex pipiens mosquitoes but differs from other mosquito densoviruses in terms of genome structure and sequence identity. Its transmission from host to host, the nature of its interactions with both its host and host's endosymbiotic bacteria Wolbachia are not known. Here, we report the presence of CpDV in the ovaries and eggs of Cx. pipiens mosquitoes in close encounters with Wolbachia. In the ovaries, CpDV amount significantly differed between mosquito lines harbouring different strains of Wolbachia and these differences were not linked to variations in Wolbachia densities. CpDV was vertically transmitted in all laboratory lines to 17%-20% of the offspring. For some females, however, the vertical transmission reached 90%. Antibiotic treatment that cured the host from Wolbachia significantly decreased both CpDV quantity and vertical transmission suggesting an impact of host microbiota, including Wolbachia, on CpDV transmission. Overall our results show that CpDV is transmitted vertically via transovarian path along with Wolbachia with which it shares the same cells. Our results are primordial to understand the dynamics of densovirus infection, their persistence and spread in populations considering their potential use in the regulation of mosquito vector populations.}, } @article {pmid30583749, year = {2018}, author = {Kittayapong, P and Kaeothaisong, NO and Ninphanomchai, S and Limohpasmanee, W}, title = {Combined sterile insect technique and incompatible insect technique: sex separation and quality of sterile Aedes aegypti male mosquitoes released in a pilot population suppression trial in Thailand.}, journal = {Parasites & vectors}, volume = {11}, number = {Suppl 2}, pages = {657}, pmid = {30583749}, issn = {1756-3305}, mesh = {Aedes/microbiology/*physiology/radiation effects ; Animals ; Female ; Infertility, Male ; Longevity ; Male ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/*physiology/radiation effects ; Pilot Projects ; Population Control ; Pupa ; Sex Characteristics ; Sex Determination Analysis ; Thailand ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The sterile insect technique (SIT), which is based on irradiation-induced sterility, and incompatible insect technique (IIT), which is based on Wolbachia-induced cytoplasmic incompatibility (a kind of male sterility), have been used as alternative methods to reduce mosquito vector populations. Both methods require the release of males to reduce fertile females and suppress the number of natural populations. Different techniques of sex separation to obtain only males have been investigated previously. Our work involves an application of mechanical larval-pupal glass separators to separate Wolbachia-infected Aedes aegypti males from females at the pupal stage, prior to irradiation, and for use in a pilot field release and to assess the quality of males and females before and after sex separation and sterilization.

RESULTS: This study was the first to demonstrate the efficiency of mechanical glass separators in separating males for use in an Ae. aegypti suppression trial by a combined SIT/IIT approach. Our results indicated that male and female pupae of Wolbachia-infected Ae. aegypti mosquitoes were significantly different (p < 0.05) in weight, size, and emergence-time, which made it easier for sex separation by this mechanical method. During the pilot field release, the percentage of female contamination was detected to be quite low and significantly different between the first (0.10 ± 0.13) and the second (0.02 ± 0.02) twelve-week period. Both males and females were almost completely sterile after exposure to 70 Gy irradiation dose. We observed that both irradiated Wolbachia-infected males and females survived and lived longer than two weeks, but males could live longer than females (p < 0.05) when they were irradiated at the same irradiation dose. When comparing irradiated mosquitoes with non-irradiated ones, there was no significant difference in longevity and survival-rate between those males, but non-irradiated females lived longer than irradiated ones (p < 0.05).

CONCLUSION: Mechanical sex separation by using a larval-pupal glass separator was practically applied to obtain only males for further sterilization and open field release in a pilot population suppression trial of Ae. aegypti in Thailand. Female contamination was detected to be quite low, and skilled personnel can reduce the risk for female release. The irradiated Wolbachia-infected females accidentally released were found to be completely sterile, with shorter life span than males.}, } @article {pmid30583743, year = {2018}, author = {Moretti, R and Marzo, GA and Lampazzi, E and Calvitti, M}, title = {Cytoplasmic incompatibility management to support Incompatible Insect Technique against Aedes albopictus.}, journal = {Parasites & vectors}, volume = {11}, number = {Suppl 2}, pages = {649}, pmid = {30583743}, issn = {1756-3305}, mesh = {Aedes/*microbiology ; Animals ; Cytoplasm/microbiology ; Female ; Fertility ; Male ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology ; Population Control ; Risk Assessment ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The transinfection of the endosymbiotic bacterium Wolbachia provides a method to produce functionally sterile males to be used to suppress mosquito vectors. ARwP is a wPip Wolbachia infected Aedes albopictus which exhibits a bidirectional incompatibility pattern with wild-types. We coupled a modelistic approach with laboratory experiments to test ARwP as a control tool and evaluate the possible occurrence of population replacement following the release of ARwP females in a wild-type (SANG) population. Repeated male-only releases were simulated and tested in the laboratory in comparison with releases contaminated with 1% ARwP females. Model simulations also investigated how migration affects the outcome of contaminated releases. Finally, the mean level of egg fertility and the long-term evolution of populations constituted by two Wolbachia infection types were studied by testing SANG and ARwP Ae. albopictus and performing more general model simulations.

RESULTS: The model was parametrized with laboratory data and simulations were compared with results of biological trials. Small populations of ARwP males and females were theoretically and experimentally demonstrated to rapidly become extinct when released in larger SANG populations. Male-only releases at a 5:1 ratio with respect to the wild-type males led to a complete suppression of the SANG population in a few generations. Contaminated releases were efficient as well but led to population replacement in the long term, when the wild-type population approached eradication. Migration significantly contrasted this trend as a 5% population turnover was sufficient to avoid any risk of population replacement. At equal frequencies between ARwP and SANG individuals, the mean egg fertility of the overall population was more than halved and suppression was self-sustaining until one of the two infection types extinguished.

CONCLUSIONS: In the case of bidirectional incompatibility patterns, the repeated release of incompatible males with small percentages of contaminant females could lead to population replacement in confined environments while it could be managed to target high efficiency and sustainability in wild-type suppression when systems are open to migration. This possibility is discussed based on various contexts and taking into consideration the possibility of integration with other control methods such as SIT and the use of larvicides.}, } @article {pmid30583288, year = {2019}, author = {Parry, R and Bishop, C and De Hayr, L and Asgari, S}, title = {Density-dependent enhanced replication of a densovirus in Wolbachia-infected Aedes cells is associated with production of piRNAs and higher virus-derived siRNAs.}, journal = {Virology}, volume = {528}, number = {}, pages = {89-100}, doi = {10.1016/j.virol.2018.12.006}, pmid = {30583288}, issn = {1096-0341}, mesh = {Aedes/cytology/*microbiology/*virology ; Animals ; Cell Line ; Cytoplasm/virology ; DNA Replication ; Densovirus/*physiology ; *Host Microbial Interactions ; RNA, Small Interfering/*genetics ; RNA, Viral/genetics ; *Virus Replication ; Wolbachia/physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia pipientis has been shown to restrict a range of RNA viruses in Drosophila melanogaster and transinfected dengue mosquito, Aedes aegypti. Here, we show that Wolbachia infection enhances replication of Aedes albopictus densovirus (AalDNV-1), a single stranded DNA virus, in Aedes cell lines in a density-dependent manner. Analysis of previously produced small RNAs of Aag2 cells showed that Wolbachia-infected cells produced greater absolute abundance of virus-derived short interfering RNAs compared to uninfected cells. Additionally, we found production of virus-derived PIWI-like RNAs (vpiRNA) produced in response to AalDNV-1 infection. Nuclear fractions of Aag2 cells produced a primary vpiRNA signature U1 bias whereas the typical "ping-pong" signature (U1 - A10) was evident in vpiRNAs from the cytoplasmic fractions. This is the first report of the density-dependent enhancement of DNA viruses by Wolbachia. Further, we report the generation of vpiRNAs in a DNA virus-host interaction for the first time.}, } @article {pmid30583042, year = {2019}, author = {Elias-Costa, AJ and Confalonieri, VA and Lanteri, AA and Rodriguero, MS}, title = {Game of clones: Is Wolbachia inducing speciation in a weevil with a mixed reproductive mode?.}, journal = {Molecular phylogenetics and evolution}, volume = {133}, number = {}, pages = {42-53}, doi = {10.1016/j.ympev.2018.12.027}, pmid = {30583042}, issn = {1095-9513}, mesh = {Animals ; Argentina ; Biological Evolution ; Brazil ; Cell Nucleus/genetics ; Female ; Genetic Speciation ; Male ; Mitochondria/genetics ; *Parthenogenesis ; Reproduction ; Weevils/classification/genetics/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Parthenogenesis is widely distributed in Metazoa but it is especially frequent in weevils (Coleoptera, Curculionidae) with one fifth of all known cases. Previous studies have shown that in the tribe Naupactini parthenogenetic reproduction most likely originated with an infection of the endoparasitic bacterium Wolbachia pipientis. In particular, Pantomorus postfasciatus possess a mixed reproductive mode: some populations have males while in others they are absent, and females produce clones by thelytoky. To better understand this scenario, we studied the population structure and infection status in 64 individuals of P. postfasciatus from Argentina and Brazil. We sequenced two mitochondrial (COI and COII) and one nuclear (ITS-1) fragments and obtained two very divergent haplogroups, one corresponding to the sexual populations uninfected with Wolbachia, and another conforming a monophyletic parthenogenetic (or presumptively parthenogenetic) and infected clade. Each of these haplogroups was identified as an independently evolutionary unit by all species delimitation analyses accomplished: multilocus *BEAST and BP&P, and single locus GMYC and K/θ rule. Additionally, present evidence suggests that Wolbachia infection occurred at least twice in all-female populations of P. postfasciatus with two different bacterial strains. Speciation mediated by Wolbachia is a recently described phenomenon and the case of P. postfasciatus is the first known case in a diplo-diploid insect. A model that describes how thelytoky-inducing phenotypes of Wolbachia could generate new lineages is discussed.}, } @article {pmid30581476, year = {2018}, author = {Niang, EHA and Bassene, H and Fenollar, F and Mediannikov, O}, title = {Biological Control of Mosquito-Borne Diseases: The Potential of Wolbachia-Based Interventions in an IVM Framework.}, journal = {Journal of tropical medicine}, volume = {2018}, number = {}, pages = {1470459}, pmid = {30581476}, issn = {1687-9686}, abstract = {People living in the tropical and subtropical regions of the world face an enormous health burden due to mosquito-borne diseases such as malaria, dengue fever, and filariasis. Historically and today, targeting mosquito vectors with, primarily, insecticide-based control strategies have been a key control strategy against major mosquito-borne diseases. However, the success to date of such approaches is under threat from multiple insecticide resistance mechanisms while vector control (VC) options are still limited. The situation therefore requires the development of innovative control measures against major mosquito-borne diseases. Transinfecting mosquitos with symbiotic bacteria that can compete with targeted pathogens or manipulate host biology to reduce their vectorial capacity are a promising and innovative biological control approach. In this review, we discuss the current state of knowledge about the association between mosquitoes and Wolbachia, emphasizing the limitations of different mosquito control strategies and the use of mosquitoes' commensal microbiota as innovative approaches to control mosquito-borne diseases.}, } @article {pmid30564272, year = {2018}, author = {Martínez-Rodríguez, P and Bella, JL}, title = {Chorthippus parallelus and Wolbachia: Overlapping Orthopteroid and Bacterial Hybrid Zones.}, journal = {Frontiers in genetics}, volume = {9}, number = {}, pages = {604}, pmid = {30564272}, issn = {1664-8021}, abstract = {Wolbachia is a well-known endosymbiotic, strictly cytoplasmic bacterium. It establishes complex cytonuclear relations that are not necessarily deleterious to its host, but that often result in reproductive alterations favoring bacterial transmission. Among these alterations, a common one is the cytoplasmic incompatibility (CI) that reduces the number of descendants in certain crosses between infected and non-infected individuals. This CI induced by Wolbachia appears in the hybrid zone that the grasshoppers Chorthippus parallelus parallelus (Cpp) and C. p. erythropus (Cpe) form in the Pyrenees: a reputed model in evolutionary biology. However, this cytonuclear incompatibility is the result of sophisticated processes of the co-divergence of the genomes of the bacterial strains and the host after generations of selection and coevolution. Here we show how these genome conflicts have resulted in a finely tuned adjustment of the bacterial strain to each pure orthopteroid taxon, and the striking appearance of another, newly identified recombinant Wolbachia strain that only occurs in hybrid grasshoppers. We propose the existence of two superimposed hybrid zones: one organized by the grasshoppers, which overlaps with a second, bacterial hybrid zone. The two hybrid zones counterbalance one another and have evolved together since the origin of the grasshopper's hybrid zone.}, } @article {pmid30560551, year = {2019}, author = {Maleki-Ravasan, N and Akhavan, N and Raz, A and Jafari, M and Zakeri, S and Dinparast Djadid, N}, title = {Co-occurrence of pederin-producing and Wolbachia endobacteria in Paederus fuscipes Curtis, 1840 (Coleoptera: Staphilinidae) and its evolutionary consequences.}, journal = {MicrobiologyOpen}, volume = {8}, number = {7}, pages = {e00777}, pmid = {30560551}, issn = {2045-8827}, abstract = {The dual occurrence of Pseudomonas-like and Wolbachia endobacteria has not been investigated in the Pederus beetles yet. We investigated pederin-producing bacteria (PPB) infection in Paederus fuscipes specimens from the southern margins of the Caspian Sea by designed genus-specific (OprF) and species-specific (16S rRNA) primers. Wolbachia infection was studied through a nested-PCR assay of Wolbachia surface protein (wsp) gene. Of the 125 analyzed beetles, 42 females (82.35%) and 15 males (20.27%) were positive to PPB infection; this is the first study reporting male P. fuscipes infection to PPB. Wolbachia infection was found in 45 female (88.23%) and 50 male (67.57%) analyzed beetles. Surprisingly, a number of 36 females (70.59%) and 13 males (17.57%) were found to be infected with both PPB and Wolbachia endosymbionts. In general, population infection rates to PPB and Wolbachia were determined to be 45.6% and 76%, respectively. The infection rates of female beetles to PPB and PPB-Wolbachia were significantly higher than males. In Paederus species, only female beetles shelter PPB and the discovery of this bacterium in adult males may reflect their cannibalistic behavior on the contaminated stages. Phylogenetic analysis showed that the sequences of OprF gene were unique among Pseudomonas spp.; however, sequences of 16S rRNA gene were related to the PPB of Pederus species. The co-occurrence and random distribution of these endobacteria may imply putative tripartite interactions among PPB, Wolbachia, and Paederus. In order to elucidate these possible tripartite interactions, further studies are required even at gender level.}, } @article {pmid30552191, year = {2019}, author = {Asselin, AK and Villegas-Ospina, S and Hoffmann, AA and Brownlie, JC and Johnson, KN}, title = {Contrasting Patterns of Virus Protection and Functional Incompatibility Genes in Two Conspecific Wolbachia Strains from Drosophila pandora.}, journal = {Applied and environmental microbiology}, volume = {85}, number = {5}, pages = {}, pmid = {30552191}, issn = {1098-5336}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animal Diseases/microbiology ; Animals ; Bacterial Proteins/genetics/metabolism ; Base Sequence ; Cytoplasm/physiology ; DNA, Bacterial/genetics/isolation & purification ; Dicistroviridae/genetics/metabolism/pathogenicity ; Drosophila/*microbiology/*virology ; Female ; Genes, Bacterial/genetics ; Genes, Viral ; Host-Pathogen Interactions ; Male ; Phenotype ; *Reproduction ; Symbiosis ; Wolbachia/genetics/*physiology/*virology ; }, abstract = {Wolbachia infections can present different phenotypes in hosts, including different forms of reproductive manipulation and antiviral protection, which may influence infection dynamics within host populations. In populations of Drosophila pandora two distinct Wolbachia strains coexist, each manipulating host reproduction: strain wPanCI causes cytoplasmic incompatibility (CI), whereas strain wPanMK causes male killing (MK). CI occurs when a Wolbachia-infected male mates with a female not infected with a compatible type of Wolbachia, leading to nonviable offspring. wPanMK can rescue wPanCI-induced CI but is unable to induce CI. The antiviral protection phenotypes provided by the wPanCI and wPanMK infections were characterized; the strains showed differential protection phenotypes, whereby cricket paralysis virus (CrPV)-induced mortality was delayed in flies infected with wPanMK but enhanced in flies infected with wPanCI compared to their respective Wolbachia-cured counterparts. Homologs of the cifA and cifB genes involved in CI identified in wPanMK and wPanCI showed a high degree of conservation; however, the CifB protein in wPanMK is truncated and is likely nonfunctional. The presence of a likely functional CifA in wPanMK and wPanMK's ability to rescue wPanCI-induced CI are consistent with the recent confirmation of CifA's involvement in CI rescue, and the absence of a functional CifB protein further supports its involvement as a CI modification factor. Taken together, these findings indicate that wPanCI and wPanMK have different relationships with their hosts in terms of their protective and CI phenotypes. It is therefore likely that different factors influence the prevalence and dynamics of these coinfections in natural Drosophila pandora hosts.IMPORTANCEWolbachia strains are common endosymbionts in insects, with multiple strains often coexisting in the same species. The coexistence of multiple strains is poorly understood but may rely on Wolbachia organisms having diverse phenotypic effects on their hosts. As Wolbachia is increasingly being developed as a tool to control disease transmission and suppress pest populations, it is important to understand the ways in which multiple Wolbachia strains persist in natural populations and how these might then be manipulated. We have therefore investigated viral protection and the molecular basis of cytoplasmic incompatibility in two coexisting Wolbachia strains with contrasting effects on host reproduction.}, } @article {pmid30551145, year = {2019}, author = {Hellemans, S and Kaczmarek, N and Marynowska, M and Calusinska, M and Roisin, Y and Fournier, D}, title = {Bacteriome-associated Wolbachia of the parthenogenetic termite Cavitermes tuberosus.}, journal = {FEMS microbiology ecology}, volume = {95}, number = {2}, pages = {}, doi = {10.1093/femsec/fiy235}, pmid = {30551145}, issn = {1574-6941}, mesh = {Animals ; Bacterial Proteins/genetics ; Brazil ; French Guiana ; Gastrointestinal Microbiome/*genetics ; Isoptera/*microbiology ; Parthenogenesis ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Symbiosis/*physiology ; Transaminases/genetics ; Trinidad and Tobago ; Wolbachia/genetics/*metabolism ; }, abstract = {Wolbachia has deeply shaped the ecology and evolution of many arthropods, and interactions between the two partners are a continuum ranging from parasitism to mutualism. Non-dispersing queens of the termite Cavitermes tuberosus are parthenogenetically produced through gamete duplication, a mode of ploidy restoration generally induced by Wolbachia. These queens display a bacteriome-like structure in the anterior part of the mesenteron. Our study explores the possibility of a nutritional mutualistic, rather than a parasitic, association between Wolbachia and C. tuberosus. We found a unique strain (wCtub), nested in the supergroup F, in 28 nests collected in French Guiana, the island of Trinidad and the state of Paraíba, Brazil (over 3500 km). wCtub infects individuals regardless of caste, sex or reproductive (sexual versus parthenogenetic) origin. qPCR assays reveal that Wolbachia densities are higher in the bacteriome-like structure and in the surrounding gut compared to other somatic tissues. High-throughput 16S rRNA gene amplicon sequencing reveals that Wolbachia represents over 97% of bacterial reads present in the bacteriome structure. BLAST analyses of 16S rRNA, bioA (a gene of the biosynthetic pathway of B vitamins) and five multilocus sequence typing genes indicated that wCtub shares 99% identity with wCle, an obligate nutritional mutualist of the bedbug Cimex lectularius.}, } @article {pmid30545384, year = {2018}, author = {Baldini, F and Rougé, J and Kreppel, K and Mkandawile, G and Mapua, SA and Sikulu-Lord, M and Ferguson, HM and Govella, N and Okumu, FO}, title = {First report of natural Wolbachia infection in the malaria mosquito Anopheles arabiensis in Tanzania.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {635}, pmid = {30545384}, issn = {1756-3305}, support = {WT102350/Z/13/WT_/Wellcome Trust/United Kingdom ; EP-C-15-008/EPA/EPA/United States ; /WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Anopheles/classification/*microbiology ; Cluster Analysis ; DNA, Bacterial/genetics ; Female ; Genetic Variation ; Malaria/*transmission ; Mosquito Vectors/classification/*microbiology ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Tanzania ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Natural infections of the endosymbiont bacteria Wolbachia have recently been discovered in populations of the malaria mosquito Anopheles gambiae (s.l.) in Burkina Faso and Mali, West Africa. This Anopheles specific strain wAnga limits the malaria parasite Plasmodium falciparum infections in the mosquito, thus it offers novel opportunities for malaria control.

RESULTS: We investigated Wolbachia presence in Anopheles arabiensis and Anopheles funestus, which are the two main malaria vectors in the Kilombero Valley, a malaria endemic region in south-eastern Tanzania. We found 3.1% (n = 65) and 7.5% (n = 147) wAnga infection prevalence in An. arabiensis in mosquitoes collected in 2014 and 2016, respectively, while no infection was detected in An. funestus (n = 41). Phylogenetic analysis suggests that at least two distinct strains of wAnga were detected, both belonging to Wolbachia supergroup A and B.

CONCLUSIONS: To our knowledge, this is the first confirmation of natural Wolbachia in malaria vectors in Tanzania, which opens novel questions on the ecological and genetic basis of its persistence and pathogen transmission in the vector hosts. Understanding the basis of interactions between Wolbachia, Anopheles mosquitoes and malaria parasites is crucial for investigation of its potential application as a biocontrol strategy to reduce malaria transmission, and assessment of how natural wAnga infections influence pathogen transmission in different ecological settings.}, } @article {pmid30541614, year = {2018}, author = {Kanté, ST and Melachio, T and Ofon, E and Njiokou, F and Simo, G}, title = {Detection of Wolbachia and different trypanosome species in Glossina palpalis palpalis populations from three sleeping sickness foci of southern Cameroon.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {630}, pmid = {30541614}, issn = {1756-3305}, mesh = {Animals ; Cameroon/epidemiology ; Female ; Humans ; Male ; Polymorphism, Genetic ; Symbiosis ; Trypanosoma/classification/genetics/*isolation & purification/physiology ; Trypanosomiasis, African/epidemiology/*parasitology/transmission ; Tsetse Flies/*microbiology/*parasitology/physiology ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {BACKGROUND: African trypanosomiases are caused by trypanosomes that are cyclically transmitted by tsetse. Investigations aiming to generate knowledge on the bacterial fauna of tsetse have revealed distinct symbiotic microorganisms. Furthermore, studies addressing the tripartite association between trypanosomes-tsetse-symbionts relationship have so far been contradictory. Most studies included Sodalis glossinudius and, consequently, the association involving Wolbachia is poorly understood. Understanding the vectorial competence of tsetse requires decrypting these tripartite associations. In this study, we identified Wolbachia and trypanosomes in Glossina palpalis palpalis from three human African trypanosomiasis (HAT) foci in southern Cameroon.

METHODS: Tsetse flies were captured with pyramidal traps in the Bipindi, Campo and Fontem HAT foci. After morphological identification, DNA was extracted from whole tsetse flies and Wolbachia and trypanosomes were identified by PCR using different trypanosome-specific primers and two Wolbachia-specific primers (Wolbachia surface protein and 16S rRNA genes). Statistical analyses were performed to compare the trypanosome and Wolbachia infection rates between villages and different foci and to look for an association between these microorganisms.

RESULTS: From a total of 2122 tsetse flies, 790 G. p. palpalis were analyzed. About 25.32% of flies hosted Wolbachia and 31.84% of non-teneral flies were infected by at least one trypanosome species. There was no significant difference between the global Wolbachia prevalence revealed by the two markers while some differences were observed between HAT foci. From 248 G. p. palpalis with trypanosome infections, 62.90% were with T. vivax, 34.68% with T. congolense forest, 16.13% with T. brucei (s.l.) and 2.42% with T. congolense savannah. Of all trypanosome-infected flies, 29.84% hosted Wolbachia and no association was observed between Wolbachia and trypanosome co-infections.

CONCLUSIONS: This study revealed differences in the prevalence of Wolbachia and trypanosomes in G. p. palpalis according to HAT foci. The use of only one marker has underestimated the prevalence of Wolbachia, thus more markers in subsequent studies may improve its detection. The presence of Wolbachia seems to have no impact on the establishment of trypanosomes in G. p. palpalis. The tripartite association between tsetse, Wolbachia and trypanosomes varies according to studied areas. Studies aiming to evaluate the genetic polymorphism of Wolbachia and its density in tsetse flies could help to better understand this association.}, } @article {pmid30483601, year = {2018}, author = {Jeffries, CL and Lawrence, GG and Golovko, G and Kristan, M and Orsborne, J and Spence, K and Hurn, E and Bandibabone, J and Tantely, LM and Raharimalala, FN and Keita, K and Camara, D and Barry, Y and Wat'senga, F and Manzambi, EZ and Afrane, YA and Mohammed, AR and Abeku, TA and Hedge, S and Khanipov, K and Pimenova, M and Fofanov, Y and Boyer, S and Irish, SR and Hughes, GL and Walker, T}, title = {Novel Wolbachia strains in Anopheles malaria vectors from Sub-Saharan Africa.}, journal = {Wellcome open research}, volume = {3}, number = {}, pages = {113}, pmid = {30483601}, issn = {2398-502X}, support = {U01CK000512/ACL/ACL HHS/United States ; R21 AI124452/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; R01 AI123074/AI/NIAID NIH HHS/United States ; }, abstract = {Background: Wolbachia, a common insect endosymbiotic bacterium that can influence pathogen transmission and manipulate host reproduction, has historically been considered absent from the Anopheles (An.) genera, but has recently been found in An. gambiae s.l. populations in West Africa. As there are numerous Anopheles species that have the capacity to transmit malaria, we analysed a range of species across five malaria endemic countries to determine Wolbachia prevalence rates, characterise novel Wolbachia strains and determine any correlation between the presence of Plasmodium, Wolbachia and the competing bacterium Asaia. Methods: Anopheles adult mosquitoes were collected from five malaria-endemic countries: Guinea, Democratic Republic of the Congo (DRC), Ghana, Uganda and Madagascar, between 2013 and 2017. Molecular analysis was undertaken using quantitative PCR, Sanger sequencing, Wolbachia multilocus sequence typing (MLST) and high-throughput amplicon sequencing of the bacterial 16S rRNA gene. Results: Novel Wolbachia strains were discovered in five species: An. coluzzii, An. gambiae s.s., An. arabiensis, An. moucheti and An. species A, increasing the number of Anopheles species known to be naturally infected. Variable prevalence rates in different locations were observed and novel strains were phylogenetically diverse, clustering with Wolbachia supergroup B strains. We also provide evidence for resident strain variants within An. species A. Wolbachia is the dominant member of the microbiome in An. moucheti and An. species A but present at lower densities in An. coluzzii. Interestingly, no evidence of Wolbachia/Asaia co-infections was seen and Asaia infection densities were shown to be variable and location dependent. Conclusions: The important discovery of novel Wolbachia strains in Anopheles provides greater insight into the prevalence of resident Wolbachia strains in diverse malaria vectors. Novel Wolbachia strains (particularly high-density strains) are ideal candidate strains for transinfection to create stable infections in other Anopheles mosquito species, which could be used for population replacement or suppression control strategies.}, } @article {pmid30535314, year = {2019}, author = {Leimeister, CA and Schellhorn, J and Dörrer, S and Gerth, M and Bleidorn, C and Morgenstern, B}, title = {Prot-SpaM: fast alignment-free phylogeny reconstruction based on whole-proteome sequences.}, journal = {GigaScience}, volume = {8}, number = {3}, pages = {}, pmid = {30535314}, issn = {2047-217X}, mesh = {Amino Acid Sequence ; Animals ; Bacteria/classification ; Databases, Protein ; *Phylogeny ; Plants/classification ; Proteome/*chemistry ; Sequence Alignment/*methods ; *Software ; }, abstract = {Word-based or 'alignment-free' sequence comparison has become an active research area in bioinformatics. While previous word-frequency approaches calculated rough measures of sequence similarity or dissimilarity, some new alignment-free methods are able to accurately estimate phylogenetic distances between genomic sequences. One of these approaches is Filtered Spaced Word Matches. Here, we extend this approach to estimate evolutionary distances between complete or incomplete proteomes; our implementation of this approach is called Prot-SpaM. We compare the performance of Prot-SpaM to other alignment-free methods on simulated sequences and on various groups of eukaryotic and prokaryotic taxa. Prot-SpaM can be used to calculate high-quality phylogenetic trees for dozens of whole-proteome sequences in a matter of seconds or minutes and often outperforms other alignment-free approaches. The source code of our software is available through Github: https://github.com/jschellh/ProtSpaM.}, } @article {pmid30535231, year = {2019}, author = {Qi, LD and Sun, JT and Hong, XY and Li, YX}, title = {Diversity and Phylogenetic Analyses Reveal Horizontal Transmission of Endosymbionts Between Whiteflies and Their Parasitoids.}, journal = {Journal of economic entomology}, volume = {112}, number = {2}, pages = {894-905}, doi = {10.1093/jee/toy367}, pmid = {30535231}, issn = {1938-291X}, mesh = {Animals ; China ; *Hemiptera ; Phylogeny ; Symbiosis ; *Wolbachia ; }, abstract = {Endosymbionts are widely distributed among insects via intraspecific vertical transmission and interspecific horizontal transmission. Parasitoids have attracted considerable interest due to their possible role in the horizontal transmission of endosymbionts. Horizontal transmission of endosymbionts between whiteflies via parasitoids has been revealed in the laboratory. However, whether this occurs under field conditions remains unknown. Here, the diversity and phylogenetic relationships of endosymbionts in 1,350 whiteflies and 36 parasitoids that emerged from whitefly nymphs collected from three locations in Jiangsu Province of China were investigated. Only Rickettsia and Wolbachia were identified in both whiteflies and parasitoids, with an overall infection frequency of 22.67% in whiteflies and 16.67% in parasitoids for Wolbachia and of 12.15% in whiteflies and 25% in parasitoids for Rickettsia. Despite the distant relationship between whiteflies and their parasitoids, phylogenetic analyses revealed that the Rickettsia and Wolbachia individuals collected from the two types of organisms were grouped together. Furthermore, shared haplotypes were also identified, which was consistent with the horizontal transmission of endosymbionts between parasitoids and whiteflies. In addition, a parasitoid resistance-related symbiont, Hamiltonella, was detected in whiteflies at a 100% infection frequency, probably accounting for the relatively low parasitism of the whiteflies in the field. The factors affecting the infection frequency of the four secondary endosymbionts in whiteflies were also examined.}, } @article {pmid30534598, year = {2018}, author = {Chung, M and Munro, JB and Tettelin, H and Dunning Hotopp, JC}, title = {Using Core Genome Alignments To Assign Bacterial Species.}, journal = {mSystems}, volume = {3}, number = {6}, pages = {}, pmid = {30534598}, issn = {2379-5077}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {With the exponential increase in the number of bacterial taxa with genome sequence data, a new standardized method to assign species designations is needed that is consistent with classically obtained taxonomic analyses. This is particularly acute for unculturable, obligate intracellular bacteria with which classically defined methods, like DNA-DNA hybridization, cannot be used, such as those in the Rickettsiales. In this study, we generated nucleotide-based core genome alignments for a wide range of genera with classically defined species, as well as those within the Rickettsiales. We created a workflow that uses the length, sequence identity, and phylogenetic relationships inferred from core genome alignments to assign genus and species designations that recapitulate classically obtained results. Using this method, most classically defined bacterial genera have a core genome alignment that is ≥10% of the average input genome length. Both Anaplasma and Neorickettsia fail to meet this criterion, indicating that the taxonomy of these genera should be reexamined. Consistently, genomes from organisms with the same species epithet have ≥96.8% identity of their core genome alignments. Additionally, these core genome alignments can be used to generate phylogenomic trees to identify monophyletic clades that define species and neighbor-network trees to assess recombination across different taxa. By these criteria, Wolbachia organisms are delineated into species different from the currently used supergroup designations, while Rickettsia organisms are delineated into 9 distinct species, compared to the current 27 species. By using core genome alignments to assign taxonomic designations, we aim to provide a high-resolution, robust method to guide bacterial nomenclature that is aligned with classically obtained results. IMPORTANCE With the increasing availability of genome sequences, we sought to develop and apply a robust, portable, and high-resolution method for the assignment of genera and species designations that can recapitulate classically defined taxonomic designations. Using cutoffs derived from the lengths and sequence identities of core genome alignments along with phylogenetic analyses, we sought to evaluate or reevaluate genus- and species-level designations for diverse taxa, with an emphasis on the order Rickettsiales, where species designations have been applied inconsistently. Our results indicate that the Rickettsia genus has an overabundance of species designations, that the current Anaplasma and Neorickettsia genus designations are both too broad and need to be divided, and that there are clear demarcations of Wolbachia species that do not align precisely with the existing supergroup designations.}, } @article {pmid30533772, year = {2018}, author = {Chung, M and Teigen, L and Libro, S and Bromley, RE and Kumar, N and Sadzewicz, L and Tallon, LJ and Foster, JM and Michalski, ML and Dunning Hotopp, JC}, title = {Multispecies Transcriptomics Data Set of Brugia malayi, Its Wolbachia Endosymbiont wBm, and Aedes aegypti across the B. malayi Life Cycle.}, journal = {Microbiology resource announcements}, volume = {7}, number = {18}, pages = {}, pmid = {30533772}, issn = {2576-098X}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {Here, we present a comprehensive transcriptomics data set of Brugia malayi, its Wolbachia endosymbiont wBm, and its vector host. This study samples from 16 stages across the entire B. malayi life cycle, including stage 1 through 4 larvae, adult males and females, embryos, immature microfilariae, and mature microfilariae.}, } @article {pmid30533608, year = {2018}, author = {Newton, ILG and Sheehan, KB}, title = {Gateway Entry Vector Library of Wolbachia pipientis Candidate Effectors from Strain wMel.}, journal = {Microbiology resource announcements}, volume = {7}, number = {1}, pages = {}, pmid = {30533608}, issn = {2576-098X}, abstract = {Wolbachia pipientis is an intracellular symbiont that modifies host biology using a type IV secretion system to inject bacterial effectors into the host cytoplasm. We utilized a bioinformatics approach to predict Wolbachia effectors and cloned the candidates into an entry vector, which can be utilized for subsequent analyses.}, } @article {pmid30503908, year = {2019}, author = {McLean, BJ and Dainty, KR and Flores, HA and O'Neill, SL}, title = {Differential suppression of persistent insect specific viruses in trans-infected wMel and wMelPop-CLA Aedes-derived mosquito lines.}, journal = {Virology}, volume = {527}, number = {}, pages = {141-145}, pmid = {30503908}, issn = {1096-0341}, support = {/WT_/Wellcome Trust/United Kingdom ; 102591/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Cell Line ; Coinfection ; Flavivirus/*physiology ; Microbial Interactions ; Mosquito Vectors/microbiology/virology ; Orthobunyavirus/*physiology ; RNA, Viral/genetics/metabolism ; Species Specificity ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Wolbachia suppresses the replication of +ssRNA viruses such as dengue and Zika viruses in Aedes aegypti mosquitoes. However, the range of viruses affected by this endosymbiont is yet to be explored. Recently, novel insect-specific viruses (ISVs) have been described from numerous mosquito species and mosquito-derived cell lines. Cell-fusing agent virus (Flaviviridae) and Phasi Charoen-like virus (Bunyaviridae) persistently infect the Ae. aegypti cell line Aag2 which has been used for experimental studies with both the wMel and wMelPop-CLA strains. Wolbachia was found to restrict the replication of CFAV but not the PCLV infection in these lines. Furthermore, an additional Ae. albopictus cell line (RML-12) which contained either wMel or wMelPop-CLA was assessed. While no infectious +ssRNA or dsRNA viruses were detected, a PCLV infection was identified. These observations provide additional evidence to support that insect-specific, +ssRNA viruses can be suppressed in cell culture by Wolbachia but -ssRNA viruses may not.}, } @article {pmid30514847, year = {2018}, author = {Rahimi-Kaldeh, S and Ashouri, A and Bandani, A and Ris, N}, title = {Abiotic and biotic factors influence diapause induction in sexual and asexual strains of Trichogramma brassicae (Hym: Trichogrammatidae).}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {17600}, pmid = {30514847}, issn = {2045-2322}, mesh = {Animals ; Diapause, Insect/*physiology ; Photoperiod ; Reproduction, Asexual/*physiology ; Sexual Behavior, Animal/*physiology ; Temperature ; Wasps/*growth & development ; Wolbachia/pathogenicity ; }, abstract = {The effects of some abiotic (maternal photoperiod and offspring developmental temperature) and biotic (host quality during both maternal and offspring generations) factors on diapause induction were investigated for two sympatric strains of Trichogramma brassicae Bezdenko (Hym: Trichogrammatidae) differing by infection status with regard to Wolbachia. The mode of reproduction, developmental temperature, maternal photoperiod and the quality of the host significantly affected diapause induction. The highest percentage of diapausing individuals were observed with the sexual strain reared at 14 °C using a "high quality" host and after a long photophase during the maternal generation. Environment-by-Environment, as well as Genotype-by-Environment interactions, was observed. All these patterns were discussed with the goal of providing relevant protocols for the commercial mass-rearing of such biocontrol agents. A successful diapause in beneficial insects could affect the efficiency of mass rearing by increasing the duration of storage conditions based on a high percent emergence and providing a large number of individuals at the appropriate time in the field season.}, } @article {pmid30498246, year = {2018}, author = {Jeffries, CL and Tantely, LM and Raharimalala, FN and Hurn, E and Boyer, S and Walker, T}, title = {Diverse novel resident Wolbachia strains in Culicine mosquitoes from Madagascar.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {17456}, pmid = {30498246}, issn = {2045-2322}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Alleles ; Animals ; *Biodiversity ; Culicidae/*microbiology ; DNA, Mitochondrial ; Female ; Geography ; Madagascar ; Mosquito Vectors/*microbiology ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia endosymbiotic bacteria are widespread throughout insect species and Wolbachia transinfected in Aedes mosquito species has formed the basis for biocontrol programs as Wolbachia strains inhibit arboviral replication and can spread through populations. Resident strains in wild Culicine mosquito populations (the vectors of most arboviruses) requires further investigation given resident strains can also affect arboviral transmission. As Madagascar has a large diversity of both Culicine species and has had recent arboviral outbreaks, an entomology survey was undertaken, in five ecologically diverse sites, to determine the Wolbachia prevalence. We detected diverse novel resident Wolbachia strains within the Aedeomyia, Culex, Ficalbia, Mansonia and Uranotaenia genera. Wolbachia prevalence rates and strain characterisation through Sanger sequencing with multilocus sequence typing (MLST) and phylogenetic analysis revealed significant diversity and we detected co-infections with the environmentally acquired bacteria Asaia. Mosquitoes were screened for major arboviruses to investigate if any evidence could be provided for their potential role in transmission and we report the presence of Rift Valley fever virus in three Culex species: Culex tritaeniorhynchus, Culex antennatus and Culex decens. The implications of the presence of resident Wolbachia strains are discussed and how the discovery of novel strains can be utilized for applications in the development of biocontrol strategies.}, } @article {pmid30496431, year = {2019}, author = {Sakamoto, H and Suzuki, R and Nishizawa, N and Matsuda, T and Gotoh, T}, title = {Effects of Wolbachia/Cardinium Infection on the Mitochondrial Phylogeny of Oligonychus castaneae (Acari: Tetranychidae).}, journal = {Journal of economic entomology}, volume = {112}, number = {2}, pages = {883-893}, doi = {10.1093/jee/toy354}, pmid = {30496431}, issn = {1938-291X}, mesh = {Animals ; Bacteroidetes ; DNA, Mitochondrial ; Japan ; Male ; Phylogeny ; Symbiosis ; *Tetranychidae ; *Wolbachia ; }, abstract = {A wide range of invertebrates harbor intracellular endosymbiotic bacteria. Within these endosymbionts, Wolbachia and Cardinium, have been attracting particular attention because these bacteria frequently affect the genetic structure and genetic diversity of their hosts. They cause various reproductive alterations such as cytoplasmic incompatibility, parthenogenesis induction, male-killing, and feminization. Through these alterations, they also affect the maternally inherited organelles of their hosts. Mitochondrial DNA (mtDNA) can be used for molecular phylogenetic analysis of invertebrates. However, in Wolbachia- or Cardinium-infected invertebrates, phylogenetic trees based on mtDNA are often inconsistent with those based on nuclear DNA. In the present study, we determined the Wolbachia/Cardinium infection status of 45 populations of the mite, Oligonychus castaneae Ehara & Gotoh (Acari: Tetranychidae), collected throughout Japan. Then, we compared phylogenetic trees of O. castaneae based on both the cytochrome c oxidase subunit I (COI) gene of mtDNA and the 28S rRNA gene of nuclear DNA to clarify the effects of Wolbachia and/or Cardinium infection. We found 106 Wolbachia-infected individuals and 250 Cardinium-infected individuals in a total of 450 individuals, indicating an infection rate of 79%. No double-infected individuals were observed. In the 28S tree, almost all populations formed a single group. In the COI tree, O. castaneae formed four separate groups that more closely followed Wolbachia/Cardinium infection than geographic distribution. These results strongly suggest that the endosymbionts affected mitochondrial variation of O. castaneae.}, } @article {pmid30474731, year = {2019}, author = {Hosseinzadeh, S and Shams-Bakhsh, M and Mann, M and Fattah-Hosseini, S and Bagheri, A and Mehrabadi, M and Heck, M}, title = {Distribution and Variation of Bacterial Endosymbiont and "Candidatus Liberibacter asiaticus" Titer in the Huanglongbing Insect Vector, Diaphorina citri Kuwayama.}, journal = {Microbial ecology}, volume = {78}, number = {1}, pages = {206-222}, pmid = {30474731}, issn = {1432-184X}, support = {5300-163//California Citrus Research Board/ ; 2016-70016-24779//National Institute of Food and Agriculture/ ; 8062-22410-006-00-D//Agricultural Research Service/ ; }, mesh = {Animals ; Citrus/*microbiology/parasitology ; Endophytes/genetics/isolation & purification/*physiology ; Female ; Hemiptera/*microbiology/physiology ; Insect Vectors/microbiology/physiology ; Male ; Plant Diseases/*microbiology ; Rhizobiaceae/genetics/isolation & purification/*physiology ; *Symbiosis ; Wolbachia/genetics/isolation & purification/physiology ; }, abstract = {The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is an economic insect pest in most citrus-growing regions and the vector of 'Candidatus Liberibacter asiaticus' (CLas), one of at least three known bacteria associated with Huanglongbing (HLB or citrus greening disease). D. citri harbors bacterial endosymbionts, including Wolbachia pipientis (strain Wolbachia wDi), 'Candidatus Carsonella ruddii,' and 'Candidatus Profftella armatura.' Many important functions of these bacteria can be inferred from their genome sequences, but their interactions with each other, CLas, and their D. citri host are poorly understood. In the present study, the titers of the endosymbionts in different tissues, in each sex, and in insects reared on healthy citrus (referred to as unexposed) and CLas-infected citrus (referred to as CLas-exposed) D. citri were investigated using real-time, quantitative PCR (qPCR) using two different quantitative approaches. Wolbachia and CLas were detected in all insect tissues. The titer of Wolbachia was higher in heads of CLas-exposed males as compared to unexposed males. In males and females, Wolbachia titer was highest in the Malpighian tubules. The highest titer of CLas was observed in the gut. Profftella and Carsonella titers were significantly reduced in the bacteriome of CLas-exposed males compared with that of unexposed males, but this effect was not observed in females. In ovaries of CLas-exposed females, the Profftella and Carsonella titers were increased as compared to non-exposed females. CLas appeared to influence the overall levels of the symbionts but did not drastically perturb the overall microbial community structure. In all the assessed tissues, CLas titer in males was significantly higher than that of females using absolute quantification. These data provide a better understanding of multi-trophic interactions regulating symbiont dynamics in the HLB pathosystem.}, } @article {pmid30470196, year = {2018}, author = {Doudoumis, V and Augustinos, A and Saridaki, A and Parker, A and Abd-Alla, AMM and Bourtzis, K and Tsiamis, G}, title = {Different laboratory populations similar bacterial profile? The case of Glossina palpalis gambiensis.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {148}, pmid = {30470196}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/isolation & purification ; Enterobacteriaceae/genetics ; Female ; Gastrointestinal Tract/microbiology ; *Genetic Variation ; Male ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Tsetse Flies/*microbiology ; Wigglesworthia/genetics ; Wolbachia/genetics ; }, abstract = {BACKGROUND: Microbiota plays an important role in the biology, ecology and evolution of insects including tsetse flies. The bacterial profile of 3 Glossina palpalis gambiensis laboratory colonies was examined using 16S rRNA gene amplicon sequencing to evaluate the dynamics of the bacterial diversity within and between each G. p. gambiensis colony.

RESULTS: The three G. p. gambiensis laboratory colonies displayed similar bacterial diversity indices and OTU distribution. Larval guts displayed a higher diversity when compared with the gastrointestinal tract of adults while no statistically significant differences were observed between testes and ovaries. Wigglesworthia and Sodalis were the most dominant taxa. In more detail, the gastrointestinal tract of adults was more enriched by Wigglesworthia while Sodalis were prominent in gonads. Interestingly, in larval guts a balanced co-existence between Wigglesworthia and Sodalis was observed. Sequences assigned to Wolbachia, Propionibacterium, and Providencia were also detected but to a much lesser degree. Clustering analysis indicated that the bacterial profile in G. p. gambiensis exhibits tissue tropism, hence distinguishing the gut bacterial profile from that present in reproductive organs.

CONCLUSIONS: Our results indicated that age, gender and the origin of the laboratory colonies did not significantly influence the formation of the bacterial profile, once these populations were kept under the same rearing conditions. Within the laboratory populations a tissue tropism was observed between the gut and gonadal bacterial profile.}, } @article {pmid30470190, year = {2018}, author = {Augustinos, AA and Meki, IK and Demirbas-Uzel, G and Ouédraogo, GMS and Saridaki, A and Tsiamis, G and Parker, AG and Abd-Alla, AMM and Bourtzis, K}, title = {Nuclear and Wolbachia-based multimarker approach for the rapid and accurate identification of tsetse species.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {147}, pmid = {30470190}, issn = {1471-2180}, mesh = {Animals ; DNA, Ribosomal Spacer/genetics ; Electrophoresis, Agar Gel ; Insect Vectors/*classification ; Mitochondria/genetics ; Molecular Typing/economics/*methods ; Polymerase Chain Reaction ; Symbiosis/genetics ; Tsetse Flies/*classification/*microbiology ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Tsetse flies (Diptera: Glossinidae) are solely responsible for the transmission of African trypanosomes, causative agents of sleeping sickness in humans and nagana in livestock. Due to the lack of efficient vaccines and the emergence of drug resistance, vector control approaches such as the sterile insect technique (SIT), remain the most effective way to control disease. SIT is a species-specific approach and therefore requires accurate identification of natural pest populations at the species level. However, the presence of morphologically similar species (species complexes and sub-species) in tsetse flies challenges the successful implementation of SIT-based population control.

RESULTS: In this study, we evaluate different molecular tools that can be applied for the delimitation of different Glossina species using tsetse samples derived from laboratory colonies, natural populations and museum specimens. The use of mitochondrial markers, nuclear markers (including internal transcribed spacer 1 (ITS1) and different microsatellites), and bacterial symbiotic markers (Wolbachia infection status) in combination with relatively inexpensive techniques such as PCR, agarose gel electrophoresis, and to some extent sequencing provided a rapid, cost effective, and accurate identification of several tsetse species.

CONCLUSIONS: The effectiveness of SIT benefits from the fine resolution of species limits in nature. The present study supports the quick identification of large samples using simple and cost effective universalized protocols, which can be easily applied by countries/laboratories with limited resources and expertise.}, } @article {pmid30470187, year = {2018}, author = {Ouedraogo, GMS and Demirbas-Uzel, G and Rayaisse, JB and Gimonneau, G and Traore, AC and Avgoustinos, A and Parker, AG and Sidibe, I and Ouedraogo, AG and Traore, A and Bayala, B and Vreysen, MJB and Bourtzis, K and Abd-Alla, AMM}, title = {Prevalence of trypanosomes, salivary gland hypertrophy virus and Wolbachia in wild populations of tsetse flies from West Africa.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {153}, pmid = {30470187}, issn = {1471-2180}, mesh = {Africa, Western ; Animals ; Cytomegalovirus/*isolation & purification/pathogenicity ; Geography ; Ghana ; Humans ; Insect Vectors/microbiology/parasitology/virology ; Prevalence ; Spiroplasma/isolation & purification ; Symbiosis ; Trypanosoma/*isolation & purification ; Tsetse Flies/*microbiology/*parasitology/*virology ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Tsetse flies are vectors of African trypanosomes, protozoan parasites that cause sleeping sickness (or human African trypanosomosis) in humans and nagana (or animal African trypanosomosis) in livestock. In addition to trypanosomes, four symbiotic bacteria Wigglesworthia glossinidia, Sodalis glossinidius, Wolbachia, Spiroplasma and one pathogen, the salivary gland hypertrophy virus (SGHV), have been reported in different tsetse species. We evaluated the prevalence and coinfection dynamics between Wolbachia, trypanosomes, and SGHV in four tsetse species (Glossina palpalis gambiensis, G. tachinoides, G. morsitans submorsitans, and G. medicorum) that were collected between 2008 and 2015 from 46 geographical locations in West Africa, i.e. Burkina Faso, Mali, Ghana, Guinea, and Senegal.

RESULTS: The results indicated an overall low prevalence of SGHV and Wolbachia and a high prevalence of trypanosomes in the sampled wild tsetse populations. The prevalence of all three infections varied among tsetse species and sample origin. The highest trypanosome prevalence was found in Glossina tachinoides (61.1%) from Ghana and in Glossina palpalis gambiensis (43.7%) from Senegal. The trypanosome prevalence in the four species from Burkina Faso was lower, i.e. 39.6% in Glossina medicorum, 18.08%; in Glossina morsitans submorsitans, 16.8%; in Glossina tachinoides and 10.5% in Glossina palpalis gambiensis. The trypanosome prevalence in Glossina palpalis gambiensis was lowest in Mali (6.9%) and Guinea (2.2%). The prevalence of SGHV and Wolbachia was very low irrespective of location or tsetse species with an average of 1.7% for SGHV and 1.0% for Wolbachia. In some cases, mixed infections with different trypanosome species were detected. The highest prevalence of coinfection was Trypanosoma vivax and other Trypanosoma species (9.5%) followed by coinfection of T. congolense with other trypanosomes (7.5%). The prevalence of coinfection of T. vivax and T. congolense was (1.0%) and no mixed infection of trypanosomes, SGHV and Wolbachia was detected.

CONCLUSION: The results indicated a high rate of trypanosome infection in tsetse wild populations in West African countries but lower infection rate of both Wolbachia and SGHV. Double or triple mixed trypanosome infections were found. In addition, mixed trypanosome and SGHV infections existed however no mixed infections of trypanosome and/or SGHV with Wolbachia were found.}, } @article {pmid30470186, year = {2018}, author = {Kariithi, HM and Boucias, DG and Murungi, EK and Meki, IK and Demirbaş-Uzel, G and van Oers, MM and Vreysen, MJB and Abd-Alla, AMM and Vlak, JM}, title = {Coevolution of hytrosaviruses and host immune responses.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {183}, pmid = {30470186}, issn = {1471-2180}, mesh = {Animals ; *Biological Coevolution ; Cytomegalovirus/*genetics/immunology ; DNA Viruses/genetics ; DNA, Viral/genetics ; *Evolution, Molecular ; Genome Size ; *Host Microbial Interactions ; Houseflies/immunology/virology ; Insect Viruses/genetics/immunology ; Phylogeny ; Salivary Glands/pathology/virology ; Tsetse Flies/immunology/*virology ; Virion/immunology ; Virus Replication ; }, abstract = {BACKGROUND: Hytrosaviruses (SGHVs; Hytrosaviridae family) are double-stranded DNA (dsDNA) viruses that cause salivary gland hypertrophy (SGH) syndrome in flies. Two structurally and functionally distinct SGHVs are recognized; Glossina pallidipes SGHV (GpSGHV) and Musca domestica SGHV (MdSGHV), that infect the hematophagous tsetse fly and the filth-feeding housefly, respectively. Genome sizes and gene contents of GpSGHV (~ 190 kb; 160-174 genes) and MdSGHV (~ 124 kb; 108 genes) may reflect an evolution with the SGHV-hosts resulting in differences in pathobiology. Whereas GpSGHV can switch from asymptomatic to symptomatic infections in response to certain unknown cues, MdSGHV solely infects symptomatically. Overt SGH characterizes the symptomatic infections of SGHVs, but whereas MdSGHV induces both nuclear and cellular hypertrophy (enlarged non-replicative cells), GpSGHV induces cellular hyperplasia (enlarged replicative cells). Compared to GpSGHV's specificity to Glossina species, MdSGHV infects other sympatric muscids. The MdSGHV-induced total shutdown of oogenesis inhibits its vertical transmission, while the GpSGHV's asymptomatic and symptomatic infections promote vertical and horizontal transmission, respectively. This paper reviews the coevolution of the SGHVs and their hosts (housefly and tsetse fly) based on phylogenetic relatedness of immune gene orthologs/paralogs and compares this with other virus-insect models.

RESULTS: Whereas MdSGHV is not vertically transmitted, GpSGHV is both vertically and horizontally transmitted, and the balance between the two transmission modes may significantly influence the pathogenesis of tsetse virus. The presence and absence of bacterial symbionts (Wigglesworthia and Sodalis) in tsetse and Wolbachia in the housefly, respectively, potentially contributes to the development of SGH symptoms. Unlike MdSGHV, GpSGHV contains not only host-derived proteins, but also appears to have evolutionarily recruited cellular genes from ancestral host(s) into its genome, which, although may be nonessential for viral replication, potentially contribute to the evasion of host's immune responses. Whereas MdSGHV has evolved strategies to counteract both the housefly's RNAi and apoptotic responses, the housefly has expanded its repertoire of immune effector, modulator and melanization genes compared to the tsetse fly.

CONCLUSIONS: The ecologies and life-histories of the housefly and tsetse fly may significantly influence coevolution of MdSGHV and GpSGHV with their hosts. Although there are still many unanswered questions regarding the pathogenesis of SGHVs, and the extent to which microbiota influence expression of overt SGH symptoms, SGHVs are attractive 'explorers' to elucidate the immune responses of their hosts, and the transmission modes of other large DNA viruses.}, } @article {pmid30470185, year = {2018}, author = {Schneider, DI and Parker, AG and Abd-Alla, AM and Miller, WJ}, title = {High-sensitivity detection of cryptic Wolbachia in the African tsetse fly (Glossina spp.).}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {140}, pmid = {30470185}, issn = {1471-2180}, support = {P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Female ; In Situ Hybridization, Fluorescence/*methods ; Insect Vectors/microbiology ; Limit of Detection ; Male ; Polymerase Chain Reaction/*methods ; Sensitivity and Specificity ; Symbiosis ; Tsetse Flies/*microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: In African tsetse flies Glossina, spp. detection of bacterial symbionts such as Wolbachia is challenging since their prevalence and distribution are patchy, and natural symbiont titers can range at levels far below detection limit of standard molecular techniques. Reliable estimation of symbiont infection frequency, especially with regard to interrelations between symbionts and their potential impact on host biology, is of pivotal interest in the context of future applications for the control and eradication of Glossina-vectored African trypanosomosis. The presence or absence of symbionts is routinely screened with endpoint polymerase chain reaction (PCR), which has numerous advantages, but reaches its limits, when detecting infections at natural low titer. To not only determine presence of native tsetse symbionts but also to localize them to specific host tissues, fluorescence in situ hybridization (FISH) can be applied. However, classic FISH assays may not detect low-titer infections due to limitations in sensitivity.

RESULTS: We have compared classic endpoint PCR with high-sensitivity blot-PCR. We demonstrate that the latter technique allows for clear detection of low-titer Wolbachia in the morsitans and palpalis groups while classic endpoint PCR does not. In order to localize Wolbachia in situ in high and low-titer Glossina species, we applied high-end Stellaris® rRNA-FISH. We show that with this high sensitivity method, even low amounts of Wolbachia can be traced in specific tissues. Furthermore, we highlight that more tissues and organs than previously recorded are infested with Wolbachia in subspecies of the morsitans and palpalis groups.

CONCLUSIONS: Our results demonstrate that overall symbiont infection frequencies as well as the presence in specific host tissues may be underestimated when using low-sensitivity methods. To better understand the complex interrelation of tsetse flies and their native symbionts plus the pathogenic trypanosomes, it is important to consider application of a broader range of high-sensitivity detection tools.}, } @article {pmid30470179, year = {2018}, author = {Demirbas-Uzel, G and De Vooght, L and Parker, AG and Vreysen, MJB and Mach, RL and Van Den Abbeele, J and Abd-Alla, AMM}, title = {Combining paratransgenesis with SIT: impact of ionizing radiation on the DNA copy number of Sodalis glossinidius in tsetse flies.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {160}, pmid = {30470179}, issn = {1471-2180}, mesh = {Animals ; DNA/*radiation effects ; Enterobacteriaceae/*genetics/*radiation effects ; Enterobacteriaceae Infections ; Female ; Insect Control/*methods ; Insect Vectors/microbiology ; Male ; *Radiation, Ionizing ; Symbiosis ; Tsetse Flies/*microbiology ; }, abstract = {BACKGROUND: Tsetse flies (Diptera: Glossinidae) are the cyclical vectors of the causative agents of African Trypanosomosis, which has been identified as a neglected tropical disease in both humans and animals in many regions of sub-Saharan Africa. The sterile insect technique (SIT) has shown to be a powerful method to manage tsetse fly populations when used in the frame of an area-wide integrated pest management (AW-IPM) program. To date, the release of sterile males to manage tsetse fly populations has only been implemented in areas to reduce transmission of animal African Trypanosomosis (AAT). The implementation of the SIT in areas with Human African Trypanosomosis (HAT) would require additional measures to eliminate the potential risk associated with the release of sterile males that require blood meals to survive and hence, might contribute to disease transmission. Paratransgenesis offers the potential to develop tsetse flies that are refractory to trypanosome infection by modifying their associated bacteria (Sodalis glossinidius) here after referred to as Sodalis. Here we assessed the feasibility of combining the paratransgenesis approach with SIT by analyzing the impact of ionizing radiation on the copy number of Sodalis and the vectorial capacity of sterilized tsetse males.

RESULTS: Adult Glossina morsitans morsitans that emerged from puparia irradiated on day 22 post larviposition did not show a significant decline in Sodalis copy number as compared with non-irradiated flies. Conversely, the Sodalis copy number was significantly reduced in adults that emerged from puparia irradiated on day 29 post larviposition and in adults irradiated on day 7 post emergence. Moreover, irradiating 22-day old puparia reduced the copy number of Wolbachia and Wigglesworthia in emerged adults as compared with non-irradiated controls, but the radiation treatment had no significant impact on the vectorial competence of the flies.

CONCLUSION: Although the radiation treatment significantly reduced the copy number of some tsetse fly symbionts, the copy number of Sodalis recovered with time in flies irradiated as 22-day old puparia. This recovery offers the opportunity to combine a paratransgenesis approach - using modified Sodalis to produce males refractory to trypanosome infection - with the release of sterile males to minimize the risk of disease transmission, especially in HAT endemic areas. Moreover, irradiation did not increase the vector competence of the flies for trypanosomes.}, } @article {pmid30470177, year = {2018}, author = {Kame-Ngasse, GI and Njiokou, F and Melachio-Tanekou, TT and Farikou, O and Simo, G and Geiger, A}, title = {Prevalence of symbionts and trypanosome infections in tsetse flies of two villages of the "Faro and Déo" division of the Adamawa region of Cameroon.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {159}, pmid = {30470177}, issn = {1471-2180}, mesh = {Animals ; Cameroon ; Enterobacteriaceae/*isolation & purification ; Insect Vectors/microbiology/parasitology ; Polymerase Chain Reaction ; Prevalence ; *Symbiosis ; Trypanosoma/genetics/isolation & purification ; Tsetse Flies/*microbiology/*parasitology ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Tsetse flies are vectors of human and animal African trypanosomiasis. In spite of many decades of chemotherapy and vector control, the disease has not been eradicated. Other methods like the transformation of tsetse fly symbionts to render the fly refractory to trypanosome infection are being evaluated. The aim of the present study was to evaluate the association between trypanosome infections and the presence of symbionts in these tsetse species. Tsetse flies were trapped in two villages of the "Faro and Déo" Division of the Adamawa region of Cameroon. In the field, tsetse fly species were identified and their infection by trypanosomes was checked by microscopy. In the laboratory, DNA was extracted from their midguts and the presence of symbionts (Sodalis glossinidius and Wolbachia sp.) and trypanosomes was checked by PCR. Symbionts/trypanosomes association tests were performed.

RESULTS: Three tsetse fly species including Glossina tachinoides (90.1%), Glossina morsitans submorsitans (9.4%) and Glossina fuscipes fuscipes (0.5%) were caught. In all the population we obtained an occurrence rate of 37.2% for Sodalis glossinidius and 67.6% for Wolbachia irrespective to tsetse flies species. S. glossinidius and Wolbachia sp. occurrence rates were respectively 37 and 68% for G. tachinoides and 28.6 and 59.5% for G. m. submorsitans. Between Golde Bourle and Mayo Dagoum significant differences were observed in the prevalence of symbionts. Prevalence of trypanosomes were 34.8% for Glossina tachinoides and 40.5% for Glossina morsitans submorsitans. In G. tachinoides, the trypanosome infection rates were 11, 2.6 and 13.7%, respectively, for T. brucei s.l., T. congolense forest type and T. congolense savannah type. In G. m. submorsitans, these infection rates were 16.7, 9.5 and, 2.4% respectively, for T. brucei s.l., T. congolense forest type and T. congolense savannah type.

CONCLUSIONS: The rate of tsetse fly infection by trypanosomes was low compared to those obtained in HAT foci of south Cameroon, and this rate was not statistically linked to the rate of symbiont occurrence. This study allowed to show for the first time the presence of Wolbachia sp. in the tsetse fly sub-species Glossina morsitans submorsitans and Glossina tachinoides.}, } @article {pmid30470176, year = {2018}, author = {Zaidman-Rémy, A and Vigneron, A and Weiss, BL and Heddi, A}, title = {What can a weevil teach a fly, and reciprocally? Interaction of host immune systems with endosymbionts in Glossina and Sitophilus.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {150}, pmid = {30470176}, issn = {1471-2180}, support = {R01 AI051584/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Enterobacteriaceae/immunology ; Host Microbial Interactions/*immunology ; Pest Control ; Symbiosis/*immunology ; Tsetse Flies/immunology/*microbiology ; Weevils/immunology/*microbiology ; Wigglesworthia/immunology ; Wolbachia/immunology ; }, abstract = {The tsetse fly (Glossina genus) is the main vector of African trypanosomes, which are protozoan parasites that cause human and animal African trypanosomiases in Sub-Saharan Africa. In the frame of the IAEA/FAO program 'Enhancing Vector Refractoriness to Trypanosome Infection', in addition to the tsetse, the cereal weevil Sitophilus has been introduced as a comparative system with regards to immune interactions with endosymbionts. The cereal weevil is an agricultural pest that destroys a significant proportion of cereal stocks worldwide. Tsetse flies are associated with three symbiotic bacteria, the multifunctional obligate Wigglesworthia glossinidia, the facultative commensal Sodalis glossinidius and the parasitic Wolbachia. Cereal weevils house an obligatory nutritional symbiosis with the bacterium Sodalis pierantonius, and occasionally Wolbachia. Studying insect host-symbiont interactions is highly relevant both for understanding the evolution of symbiosis and for envisioning novel pest control strategies. In both insects, the long co-evolution between host and endosymbiont has led to a stringent integration of the host-bacteria partnership. These associations were facilitated by the development of specialized host traits, including symbiont-housing cells called bacteriocytes and specific immune features that enable both tolerance and control of the bacteria. In this review, we compare the tsetse and weevil model systems and compile the latest research findings regarding their biological and ecological similarities, how the immune system controls endosymbiont load and location, and how host-symbiont interactions impact developmental features including cuticle synthesis and immune system maturation. We focus mainly on the interactions between the obligate symbionts and their host's immune systems, a central theme in both model systems. Finally, we highlight how parallel studies on cereal weevils and tsetse flies led to mutual discoveries and stimulated research on each model, creating a pivotal example of scientific improvement through comparison between relatively distant models.}, } @article {pmid30468769, year = {2019}, author = {Bi, J and Zheng, Y and Wang, RF and Ai, H and Haynes, PR and Brownlie, JC and Yu, XQ and Wang, YF}, title = {Wolbachia infection may improve learning and memory capacity of Drosophila by altering host gene expression through microRNA.}, journal = {Insect biochemistry and molecular biology}, volume = {106}, number = {}, pages = {47-54}, doi = {10.1016/j.ibmb.2018.11.007}, pmid = {30468769}, issn = {1879-0240}, mesh = {Animals ; Drosophila melanogaster/microbiology/*physiology ; Drosophila simulans/microbiology/*physiology ; *Gene Expression Regulation ; Learning ; Memory ; MicroRNAs/*genetics/metabolism ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria present in a wide range of invertebrates. Although their dramatic effects on host reproductive biology have been well studied, little is known about the effects of Wolbachia on the learning and memory capacity (LMC) of hosts, despite their distribution in the host nervous system, including brain. In this study, we found that Wolbachia infection significantly enhanced LMC in both Drosophila melanogaster and D. simulans. Expression of LMC-related genes was significantly increased in the head of D. melanogaster infected with the wMel strain, and among these genes, crebA was up-regulated the most. Knockdown of crebA in Wolbachia-infected flies significantly decreased LMC, while overexpression of crebA in Wolbachia-free flies significantly enhanced the LMC of flies. More importantly, a microRNA (miRNA), dme-miR-92b, was identified to be complementary to the 3'UTR of crebA. Wolbachia infection was correlated with reduced expression of dme-miR-92b in D. melanogaster, and dme-miR-92b negatively regulated crebA through binding to its 3'UTR region. Overexpression of dme-miR-92b in Wolbachia-infected flies by microinjection of agomirs caused a significant decrease in crebA expression and LMC, while inhibition of dme-miR-92b in Wolbachia-free flies by microinjection of antagomirs resulted in a significant increase in crebA expression and LMC. These results suggest that Wolbachia may improve LMC in Drosophila by altering host gene expression through a miRNA-target pathway. Our findings help better understand the host-endosymbiont interactions and, in particular, the impact of Wolbachia on cognitive processes in invertebrate hosts.}, } @article {pmid30459726, year = {2018}, author = {Brown, AMV and Wasala, SK and Howe, DK and Peetz, AB and Zasada, IA and Denver, DR}, title = {Comparative Genomics of Wolbachia-Cardinium Dual Endosymbiosis in a Plant-Parasitic Nematode.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {2482}, pmid = {30459726}, issn = {1664-302X}, abstract = {Wolbachia and Cardinium are among the most important and widespread of all endosymbionts, occurring in nematodes and more than half of insect and arachnid species, sometimes as coinfections. These symbionts are of significant interest as potential biocontrol agents due to their abilities to cause major effects on host biology and reproduction through cytoplasmic incompatibility, sex ratio distortion, or obligate mutualism. The ecological and metabolic effects of coinfections are not well understood. This study examined a Wolbachia-Cardinium coinfection in the plant-parasitic nematode (PPN), Pratylenchus penetrans, producing the first detailed study of such a coinfection using fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and comparative genomic analysis. Results from FISH and single-nematode PCR showed 123/127 individuals in a focal population carried Cardinium (denoted strain cPpe), and 48% were coinfected with Wolbachia strain wPpe. Both endosymbionts showed dispersed tissue distribution with highest densities in the anterior intestinal walls and gonads. Phylogenomic analyses confirmed an early place of cPpe and long distance from a sister strain in another PPN, Heterodera glycines, supporting a long history of both Cardinium and Wolbachia in PPNs. The genome of cPpe was 1.36 Mbp with 35.8% GC content, 1,131 predicted genes, 41% having no known function, and missing biotin and lipoate synthetic capacity and a plasmid present in other strains, despite having a slightly larger genome compared to other sequenced Cardinium. The larger genome revealed expansions of gene families likely involved in host-cellular interactions. More than 2% of the genes of cPpe and wPpe were identified as candidate horizontally transferred genes, with some of these from eukaryotes, including nematodes. A model of the possible Wolbachia-Cardinium interaction is proposed with possible complementation in function for pathways such as methionine and fatty acid biosynthesis and biotin transport.}, } @article {pmid30459180, year = {2019}, author = {Singh, ND}, title = {Wolbachia Infection Associated with Increased Recombination in Drosophila.}, journal = {G3 (Bethesda, Md.)}, volume = {9}, number = {1}, pages = {229-237}, pmid = {30459180}, issn = {2160-1836}, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology ; Fertility/genetics ; Genome, Insect/genetics ; Reproduction/*genetics ; Symbiosis/*genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia is a maternally-transmitted endosymbiotic bacteria that infects a large diversity of arthropod and nematode hosts. Some strains of Wolbachia are parasitic, manipulating host reproduction to benefit themselves, while other strains of Wolbachia exhibit obligate or facultative mutualisms with their host. The effects of Wolbachia on its host are many, though primarily relate to host immune and reproductive function. Here we test the hypothesis that Wolbachia infection alters the frequency of homologous recombination during meiosis. We use D. melanogaster as a model system, and survey recombination in eight wild-derived Wolbachia-infected (strain wMel) and Wolbachia-uninfected strains, controlling for genotype. We measure recombination in two intervals of the genome. Our results indicate that Wolbachia infection is associated with increased recombination in one genomic interval and not the other. The effect of Wolbachia infection on recombination is thus heterogenous across the genome. Our data also indicate a reproductive benefit of Wolbachia infection; infected females show higher fecundity than their uninfected genotypic controls. Given the prevalence of Wolbachia infection in natural populations, our findings suggest that Wolbachia infection is likely to contribute to recombination rate and fecundity variation among individuals in nature.}, } @article {pmid30456532, year = {2019}, author = {Schneider, DI and Ehrman, L and Engl, T and Kaltenpoth, M and Hua-Van, A and Le Rouzic, A and Miller, WJ}, title = {Symbiont-Driven Male Mating Success in the Neotropical Drosophila paulistorum Superspecies.}, journal = {Behavior genetics}, volume = {49}, number = {1}, pages = {83-98}, pmid = {30456532}, issn = {1573-3297}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Behavior, Animal ; Biological Evolution ; Drosophila/*microbiology ; Drosophila Proteins/metabolism ; Female ; Male ; Reproduction/*physiology ; Reproductive Isolation ; Sex Attractants/metabolism/physiology ; Sexual Behavior, Animal/physiology ; Species Specificity ; Symbiosis/*genetics ; Wolbachia/physiology ; }, abstract = {Microbial symbionts are ubiquitous associates of living organisms but their role in mediating reproductive isolation (RI) remains controversial. We addressed this knowledge gap by employing the Drosophila paulistorum-Wolbachia model system. Semispecies in the D. paulistorum species complex exhibit strong RI between each other and knockdown of obligate mutualistic Wolbachia bacteria in female D. paulistorum flies triggers loss of assortative mating behavior against males carrying incompatible Wolbachia strains. Here we set out to determine whether de novo RI can be introduced by Wolbachia-knockdown in D. paulistorum males. We show that Wolbachia-knockdown D. paulistorum males (i) are rejected as mates by wild type females, (ii) express altered sexual pheromone profiles, and (iii) are devoid of the endosymbiont in pheromone producing cells. Our findings suggest that changes in Wolbachia titer and tissue tropism can induce de novo premating isolation by directly or indirectly modulating sexual behavior of their native D. paulistorum hosts.}, } @article {pmid30449670, year = {2018}, author = {Hamilton, PT and Hodson, CN and Curtis, CI and Perlman, SJ}, title = {Genetics and Genomics of an Unusual Selfish Sex Ratio Distortion in an Insect.}, journal = {Current biology : CB}, volume = {28}, number = {23}, pages = {3864-3870.e4}, doi = {10.1016/j.cub.2018.10.035}, pmid = {30449670}, issn = {1879-0445}, support = {//CIHR/Canada ; }, mesh = {Animals ; Female ; *Genome, Insect ; Insecta/*genetics ; Male ; Repetitive Sequences, Nucleic Acid ; Reproduction ; Sex Chromosomes/*genetics ; *Sex Ratio ; Wolbachia/genetics ; }, abstract = {Diverse selfish genetic elements have evolved the ability to manipulate reproduction to increase their transmission, and this can result in highly distorted sex ratios [1]. Indeed, one of the major explanations for why sex determination systems are so dynamic is because they are shaped by ongoing coevolutionary arms races between sex-ratio-distorting elements and the rest of the genome [2]. Here, we use genetic crosses and genome analysis to describe an unusual sex ratio distortion with striking consequences on genome organization in a booklouse species, Liposcelis sp. (Insecta: Psocodea), in which two types of females coexist. Distorter females never produce sons but must mate with males (the sons of nondistorting females) to reproduce [3]. Although they are diploid and express the genes inherited from their fathers in somatic tissues, distorter females only ever transmit genes inherited from their mothers. As a result, distorter females have unusual chimeric genomes, with distorter-restricted chromosomes diverging from their nondistorting counterparts and exhibiting features of a giant non-recombining sex chromosome. The distorter-restricted genome has also acquired a gene from the bacterium Wolbachia, a well-known insect reproductive manipulator; we found that this gene has independently colonized the genomes of two other insect species with unusual reproductive systems, suggesting possible roles in sex ratio distortion in this remarkable genetic system.}, } @article {pmid30448462, year = {2019}, author = {Hu, L and Huang, M and Tang, M and Yu, J and Zheng, B}, title = {Wolbachia spread dynamics in multi-regimes of environmental conditions.}, journal = {Journal of theoretical biology}, volume = {462}, number = {}, pages = {247-258}, doi = {10.1016/j.jtbi.2018.11.009}, pmid = {30448462}, issn = {1095-8541}, mesh = {Aedes/microbiology/virology ; Animals ; *Climate ; Dengue/prevention & control/transmission ; *Models, Biological ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods ; Virus Replication ; Wolbachia/*pathogenicity ; Zika Virus Infection/prevention & control/transmission ; }, abstract = {Mosquito-borne diseases such as dengue fever and Zika kill more than 700,000 people each year in the world. A novel strategy to control these diseases employs the bacterium Wolbachia whose infection in mosquitoes blocks virus replication. The prerequisite for this measure is to release Wolbachia -infected mosquitoes to replace wild population. Due to the fluctuation of environmental conditions for mosquito growth, we develop and analyze a model of differential equations with parameters randomly changing over multiple environmental regimes. By comparing the dynamics between the stochastic system and constructed auxiliary systems, combined with other techniques, we provide sharp estimates on the threshold releasing level for Wolbachia fixation. We define the alarm period of disease transmission to measure the risk of mosquito-borne diseases. Our numerical simulations suggest that more frequent inter-regime transitions help reduce the alarm period, and the disease transmission is more sensitive to the average climatic conditions than the number of sub-regimes over a given time period. Further numerical examples also indicate that the reduction in the waiting time to suppress 95% of wild population is more evident when the releasing amount is increased up to a double of the wild population.}, } @article {pmid30448382, year = {2019}, author = {Herran, B and Cerveau, N and Houdelet, C and Bernier, C and Debenest, C and Delaunay, C and Raimond, M and Bertaux, J and Grève, P}, title = {IGFBP-rP1, a strongly conserved member of the androgenic hormone signalling pathway in Isopoda.}, journal = {General and comparative endocrinology}, volume = {272}, number = {}, pages = {9-19}, doi = {10.1016/j.ygcen.2018.11.006}, pmid = {30448382}, issn = {1095-6840}, mesh = {Androgens ; Animals ; Female ; Insulin-Like Growth Factor Binding Proteins/*genetics/*metabolism ; Isopoda/*metabolism ; Male ; Signal Transduction ; }, abstract = {The first protein which has been described to interact with the malacostracan Androgenic Gland Hormone (AGH) is a binding protein called IGFBP-rP1. It has been identified and studied in several species of decapods, in which its interaction with the masculinizing hormone and its expression patterns have been established in several ways. However, this protein remains uncharacterised to date in the other malacostracan orders, like Amphipoda and Isopoda, although they were historically the first ones in which the androgenic gland and the corresponding hormone were respectively described. In this article, we identified the IGFBP-rP1 of isopods and established its implication in the pathway of the AGH with a silencing approach in the model species Armadillidium vulgare. We also showed that this gene is expressed in all the tissues of males and females, with a similar pattern in animals infected with Wolbachia, a feminizing endosymbiont of several isopod species. The expression pattern did not differ during the development of uninfected and infected animals either. We finally studied the evolution of the IGFBP-rP1 in 68 isopod species, looking for conserved motifs and evidence of natural selection. Altogether, our results showed that this gene is constitutively expressed and strongly conserved in isopods, in which it likely constitutes a key element of the insulin/IGF signalling pathway. However, we also illustrated that IGFBP-rP1 is not sufficient on its own to explain the different developmental paths taken by the males and the females or feminized genetic males.}, } @article {pmid30440032, year = {2018}, author = {Gomes, FM and Barillas-Mury, C}, title = {Infection of anopheline mosquitoes with Wolbachia: Implications for malaria control.}, journal = {PLoS pathogens}, volume = {14}, number = {11}, pages = {e1007333}, pmid = {30440032}, issn = {1553-7374}, support = {Z01 AI000947/ImNIH/Intramural NIH HHS/United States ; Z01 AI000947/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology ; Insect Vectors ; Malaria/metabolism/*prevention & control/virology ; Wolbachia/*virology ; }, } @article {pmid30426159, year = {2019}, author = {Mohanty, I and Rath, A and Swain, SP and Pradhan, N and Hazra, RK}, title = {Wolbachia Population in Vectors and Non-vectors: A Sustainable Approach Towards Dengue Control.}, journal = {Current microbiology}, volume = {76}, number = {2}, pages = {133-143}, pmid = {30426159}, issn = {1432-0991}, support = {0//Lady Tata Memorial Trust/ ; }, mesh = {Aedes/microbiology ; Animals ; Biological Control Agents ; Communicable Disease Control/*methods ; Dengue/*prevention & control/transmission ; Female ; India ; Male ; Mosquito Vectors/classification/*microbiology ; Ovary/microbiology ; Polymerase Chain Reaction ; Prevalence ; Salivary Glands/microbiology ; Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is gram negative obligate endosymbiont known for reproductive manipulation in the host. It is important to study the presence of natural Wolbachia in mosquitoes which can later help in understanding the effect of transfected strain on indigenous strain. With this view, the present study is undertaken to focus on the prevalence, diversity, infection frequencies, phylogeny and density of indigenous Wolbachia strains in wild mosquito species of Odisha. Our study confirms Wolbachia presence in Ae. albopictus, Cx. quinquefasciatus, Cx. vishnui, Cx. gelidus, Ar. subalbatus, Mn. uniformis, and Mn. indiana. Wolbachia in the above mosquitoes were separated into two supergroups (A and B). Ae. albopictus, the major vector of dengue and chikungungunya had both super-infection and mono-infection. The ovaries of Ae. albopictus were highest in density of Wolbachia as compared to midguts or salivary glands. wAlBA and wAlbB density were variable in mosquitoes of F1 generation for both the sex and at different age. We also found that Wolbachia super-infection in females tends to increase whereas wAlbA density reduced completely as compared to wAlbB in males when they grew old. Giemsa stained squashed ovaries revealed pink pleomorphic Wolbachia cells with different shapes and forms. This study is unique in its kind covering the major aspects of the endosymbiont Wolbachia and focusing on its potential as a biocontrol agent in arboviral outbreaks. Knowledge on potential of the indigenous strain and interactions between Wolbachia and viruses can be utilized further to reduce the global burden of vector borne diseases.}, } @article {pmid30425700, year = {2018}, author = {Hubert, J and Nesvorna, M and Sopko, B and Smrz, J and Klimov, P and Erban, T}, title = {Two Populations of Mites (Tyrophagus putrescentiae) Differ in Response to Feeding on Feces-Containing Diets.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {2590}, pmid = {30425700}, issn = {1664-302X}, abstract = {Background: Tyrophagus putrescentiae is a ubiquitous mite species in soil, stored products and house dust and infests food and causes allergies in people. T. putrescentiae populations harbor different bacterial communities, including intracellular symbionts and gut bacteria. The spread of microorganisms via the fecal pellets of T. putrescentiae is a possibility that has not been studied in detail but may be an important means by which gut bacteria colonize subsequent generations of mites. Feces in soil may be a vector for the spread of microorganisms. Methods: Extracts from used mite culture medium (i.e., residual food, mite feces, and dead mite bodies) were used as a source of feces-inhabiting microorganisms as food for the mites. Two T. putrescentiae populations (L and P) were used for experiments, and they hosted the intracellular bacteria Cardinium and Wolbachia, respectively. The effects of the fecal fraction on respiration in a mite microcosm, mite nutrient contents, population growth and microbiome composition were evaluated. Results: Feces from the P population comprised more than 90% Bartonella-like sequences. Feces from the L population feces hosted Staphylococcus, Virgibacillus, Brevibacterium, Enterobacteriaceae, and Bacillus. The mites from the P population, but not the L population, exhibited increased bacterial respiration in the microcosms in comparison to no-mite controls. Both L- and P-feces extracts had an inhibitory effect on the respiration of the microcosms, indicating antagonistic interactions within feces-associated bacteria. The mite microbiomes were resistant to the acquisition of new bacterial species from the feces, but their bacterial profiles were affected. Feeding of P mites on P-feces-enriched diets resulted in an increase in Bartonella abundance from 6 to 20% of the total bacterial sequences and a decrease in Bacillus abundance. The population growth was fivefold accelerated on P-feces extracts in comparison to the control. Conclusion: The mite microbiome, to a certain extent, resists the acquisition of new bacteria when mites are fed on feces of the same species. However, a Bartonella-like bacteria-feces-enriched diet seems to be beneficial for mite populations with symbiotic Bartonella-like bacteria. Coprophagy on the feces of its own population may be a mechanism of bacterial acquisition in T. putrescentiae.}, } @article {pmid30422992, year = {2018}, author = {Grobler, Y and Yun, CY and Kahler, DJ and Bergman, CM and Lee, H and Oliver, B and Lehmann, R}, title = {Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation.}, journal = {PLoS pathogens}, volume = {14}, number = {11}, pages = {e1007445}, pmid = {30422992}, issn = {1553-7374}, support = {/Howard Hughes Medical Institute/Howard Hughes Medical Institute/United States ; P30 CA016087/CA/NCI NIH HHS/United States ; P40 OD018537/OD/NIH HHS/United States ; }, mesh = {Animals ; Culicidae ; Drosophila/genetics/microbiology ; Drosophila melanogaster/*genetics/microbiology ; Genome ; Host Microbial Interactions/*genetics ; Host-Pathogen Interactions/genetics ; Humans ; In Situ Hybridization, Fluorescence/methods ; Mosquito Vectors ; RNA Interference ; Symbiosis ; Viruses/genetics ; Whole Genome Sequencing/methods ; Wolbachia/*genetics ; }, abstract = {Wolbachia is an intracellular bacterium that infects a remarkable range of insect hosts. Insects such as mosquitos act as vectors for many devastating human viruses such as Dengue, West Nile, and Zika. Remarkably, Wolbachia infection provides insect hosts with resistance to many arboviruses thereby rendering the insects ineffective as vectors. To utilize Wolbachia effectively as a tool against vector-borne viruses a better understanding of the host-Wolbachia relationship is needed. To investigate Wolbachia-insect interactions we used the Wolbachia/Drosophila model that provides a genetically tractable system for studying host-pathogen interactions. We coupled genome-wide RNAi screening with a novel high-throughput fluorescence in situ hybridization (FISH) assay to detect changes in Wolbachia levels in a Wolbachia-infected Drosophila cell line JW18. 1117 genes altered Wolbachia levels when knocked down by RNAi of which 329 genes increased and 788 genes decreased the level of Wolbachia. Validation of hits included in depth secondary screening using in vitro RNAi, Drosophila mutants, and Wolbachia-detection by DNA qPCR. A diverse set of host gene networks was identified to regulate Wolbachia levels and unexpectedly revealed that perturbations of host translation components such as the ribosome and translation initiation factors results in increased Wolbachia levels both in vitro using RNAi and in vivo using mutants and a chemical-based translation inhibition assay. This work provides evidence for Wolbachia-host translation interaction and strengthens our general understanding of the Wolbachia-host intracellular relationship.}, } @article {pmid30416203, year = {2018}, author = {Treanor, D and Pamminger, T and Hughes, WOH}, title = {The evolution of caste-biasing symbionts in the social hymenoptera.}, journal = {Insectes sociaux}, volume = {65}, number = {4}, pages = {513-519}, pmid = {30416203}, issn = {0020-1812}, abstract = {The separation of individuals into reproductive and worker castes is the defining feature of insect societies. However, caste determination is itself a complex phenomenon, dependent on interacting genetic and environmental factors. It has been suggested by some authors that widespread maternally transmitted symbionts such as Wolbachia may be selected to interfere with caste determination, whilst others have discounted this possibility on theoretical grounds. We argue that there are in fact three distinct evolutionary scenarios in which maternally transmitted symbionts might be selected to influence the process of caste determination in a social hymenopteran host. Each of these scenarios generate testable predictions which we outline here. Given the increasing recognition of the complexity and multi-faceted nature of caste determination in social insects, we argue that maternally transmitted symbionts should also be considered as possible factors influencing the development of social hymenopterans.}, } @article {pmid30414119, year = {2019}, author = {Schuler, H and Lopez, JA and Doellman, MM and Hood, GR and Egan, SP and Feder, JL}, title = {Target-Enriched Endosymbiont Sequencing (TEEseq): A New High-Throughput Sequencing Approach Applied to the Comprehensive Characterization of Endosymbionts.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {1858}, number = {}, pages = {195-212}, doi = {10.1007/978-1-4939-8775-7_14}, pmid = {30414119}, issn = {1940-6029}, mesh = {Animals ; Bacterial Proteins/genetics ; Computational Biology/*methods ; DNA, Bacterial/analysis/genetics ; *Genome, Bacterial ; High-Throughput Nucleotide Sequencing/*methods ; Insecta/*microbiology ; Sequence Analysis, DNA/methods ; *Symbiosis ; Wolbachia/*genetics/isolation & purification/physiology ; }, abstract = {Intracellular bacteria are ubiquitous in the insect world, with perhaps the best-studied example being the alphaproteobacterium, Wolbachia. Like most endosymbionts, Wolbachia cannot be cultivated outside of its host cells, hindering traditional microbial characterization techniques. Furthermore, multiple Wolbachia strains can be present within a single host, and certain strains can be present in densities below the detection limit of current methods. To date, Wolbachia has most commonly been studied using polymerase chain reaction (PCR) amplification and Sanger DNA sequencing by targeting specific genes in the bacterium's genome. PCR amplification and Sanger sequencing of multiple Wolbachia strains requires analysis of individually cloned sequences, which is resource and labor intensive. To help mitigate these difficulties, we present a modified double digest restriction site associated DNA sequencing (ddRADseq) approach to target and sequence in parallel multiple genes by adding restriction enzyme recognition sites to gene-specific PCR primers. Adopting this strategy allows us to uniquely tag and sequence amplicons from multiple hosts simultaneously on an Illumina MiSeq platform. Our approach represents an efficient and cost-effective method to characterize multiple target genes in population surveys.}, } @article {pmid30400987, year = {2018}, author = {Niang, EHA and Bassene, H and Makoundou, P and Fenollar, F and Weill, M and Mediannikov, O}, title = {First report of natural Wolbachia infection in wild Anopheles funestus population in Senegal.}, journal = {Malaria journal}, volume = {17}, number = {1}, pages = {408}, pmid = {30400987}, issn = {1475-2875}, support = {Méditerranée Infection 10-IAHU- 03//Agence Nationale de la Recherche/ ; }, mesh = {Animals ; Anopheles/*microbiology ; Base Sequence ; Mosquito Vectors/*microbiology ; Phylogeny ; RNA, Bacterial/*analysis ; RNA, Ribosomal, 16S/*analysis ; Senegal ; Sequence Alignment ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Until very recently, Anopheles were considered naturally unable to host Wolbachia, an intracellular bacterium regarded as a potential biological control tool. Their detection in field populations of Anopheles gambiae sensu lato, suggests that they may also be present in many more anopheline species than previously thought.

RESULTS: Here, is reported the first discovery of natural Wolbachia infections in Anopheles funestus populations from Senegal, the second main malaria vector in Africa. Molecular phylogeny analysis based on the 16S rRNA gene revealed at least two Wolbachia genotypes which were named wAnfu-A and wAnfu-B, according to their close relatedness to the A and B supergroups. Furthermore, both wAnfu genotypes displayed high proximity with wAnga sequences previously described from the An. gambiae complex, with only few nucleotide differences. However, the low prevalence of infection, together with the difficulties encountered for detection, whatever method used, highlights the need to develop an effective and sensitive Wolbachia screening method dedicated to anopheline.

CONCLUSIONS: The discovery of natural Wolbachia infection in An. funestus, another major malaria vector, may overcome the main limitation of using a Wolbachia-based approach to control malaria through population suppression and/or replacement.}, } @article {pmid30391314, year = {2019}, author = {Konecka, E and Olszanowski, Z}, title = {A new Cardinium group of bacteria found in Achipteria coleoptrata (Acari: Oribatida).}, journal = {Molecular phylogenetics and evolution}, volume = {131}, number = {}, pages = {64-71}, doi = {10.1016/j.ympev.2018.10.043}, pmid = {30391314}, issn = {1095-9513}, mesh = {Animals ; Bacteroidetes/*physiology ; Base Sequence ; DNA, Ribosomal/genetics ; Likelihood Functions ; Mites/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; }, abstract = {The understanding of the biology of arthropods requires an understanding of their bacterial associates. We determined the distribution of bacteria Wolbachia sp., Rickettsia sp., Cardinium sp., Spiroplasma sp., Arsenophonus sp., Hamiltonella sp., and Flavobacterium in oribatid mites (Acari: Oribatida). We identified Cardinium sp. in Achipteria coleoptrata. This is the first report of this bacterium in A. coleoptrata. Approximately 30% of the mite population was infected by Cardinium sp. The Cardinium 16S rDNA was examined for the presence of two sequences unique for this microorganism. One of them was noted in Cardinium sp. of A. coleoptrata. In the second sequence, we found nucleotide substitution in the 7th position: A instead of T. In our opinion, this demonstrated the unique nature of Cardinium sp. of A. coleoptrata. We also determined phylogenetic relationship between Cardinium sp., including the strain found in A. coleoptrata by studying the 16S rRNA and gyrB gene sequences. It revealed that Cardinium from A. coleoptrata did not cluster together with strains from groups A, B, C or D, and constituted a separate clade E. These observations make A. coleoptrata a unique Cardinium host in terms of the distinction of the strain.}, } @article {pmid30382151, year = {2018}, author = {Rosso, F and Tagliapietra, V and Albanese, D and Pindo, M and Baldacchino, F and Arnoldi, D and Donati, C and Rizzoli, A}, title = {Reduced diversity of gut microbiota in two Aedes mosquitoes species in areas of recent invasion.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {16091}, pmid = {30382151}, issn = {2045-2322}, mesh = {Aedes/*microbiology ; Animals ; *Biodiversity ; Female ; France ; *Gastrointestinal Microbiome ; *Introduced Species ; Italy ; Phylogeny ; Principal Component Analysis ; Vietnam ; }, abstract = {Aedes mosquitoes are considered highly successful global invasive species and vectors of several pathogens of relevance for public health. Their midgut's microbiota can play an important role in affecting not only their vectorial competence but also their fitness, physiology, food digestion, metabolism, immunity and adaptation to new environmental conditions. Using high-throughput sequencing we compared the microbiota of Aedes albopictus collected in Italy with those reported in populations from France and Vietnam. We also analysed Aedes koreicus gut microbiota for the first time. We found remarkable individual difference along with common bacterial taxa in both species. Ae. albopictus collected in Italy had a lower richness and a different composition of microbiota in respect to specimens collected in France and Vietnam. It also showed a core microbiota formed mainly of bacteria of the genus Pseudomonas. Overall, the two Aedes species (Ae. albopictus and Ae. koreicus) collected in Italy, showed a large core microbiota with 75.98% of the identified Operational Taxonomic Units. Furthermore, Ae. albopictus had 2.5% prevalence of Wolbachia and 0.07% of Asaia spp, while Ae. koreicus had 14.42% of Asaia spp. and no Wolbachia. This study provides new informations on the spatial variation of the midgut bacterial communities in mosquitoes of medical relevance within areas of recent invasion and provide the basis for further studies aimed at assessing the effects of such variation on vectorial capacity for a range of pathogens.}, } @article {pmid30380317, year = {2018}, author = {Strugarek, M and Vauchelet, N and Zubelli, JP}, title = {Quantifying the survival uncertainty of Wolbachia-infected mosquitoes in a spatial model.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {15}, number = {4}, pages = {961-991}, doi = {10.3934/mbe.2018043}, pmid = {30380317}, issn = {1551-0018}, mesh = {Aedes/*microbiology ; Animals ; Chikungunya Fever/prevention & control/transmission ; Computer Simulation ; Dengue/prevention & control/transmission ; Humans ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods/statistics & numerical data ; Uncertainty ; Wolbachia/*physiology ; Zika Virus Infection/prevention & control/transmission ; }, abstract = {Artificial releases of Wolbachia-infected Aedes mosquitoes have been under study in the past yearsfor fighting vector-borne diseases such as dengue, chikungunya and zika.Several strains of this bacterium cause cytoplasmic incompatibility (CI) and can also affect their host's fecundity or lifespan, while highly reducing vector competence for the main arboviruses. We consider and answer the following questions: 1) what should be the initial condition (i.e. size of the initial mosquito population) to have invasion with one mosquito release source? We note that it is hard to have an invasion in such case. 2) How many release points does one need to have sufficiently high probability of invasion? 3) What happens if one accounts for uncertainty in the release protocol (e.g. unequal spacing among release points)? We build a framework based on existing reaction-diffusion models for the uncertainty quantification in this context,obtain both theoretical and numerical lower bounds for the probability of release successand give new quantitative results on the one dimensional case.}, } @article {pmid30374735, year = {2019}, author = {Hashmi, TR and Devi, SR and Ahmad, A and Meshram, NM and Prasad, R}, title = {Genetic Status and Endosymbionts Diversity of Bemisia tabaci (Gennadius) on Hosts Belonging to Family Malvaceae in India.}, journal = {Neotropical entomology}, volume = {48}, number = {2}, pages = {207-218}, pmid = {30374735}, issn = {1678-8052}, mesh = {Abelmoschus ; Animals ; Bacteria/*classification ; DNA Primers ; Gossypium ; Hemiptera/*microbiology ; India ; *Malvaceae ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; }, abstract = {A study was instigated to examine the genetic status and distribution of known endosymbionts namely Portiera, Rickettsia, Wolbachia, Cardinium, and Arsenophonus in the populations of Bemisia tabaci (Gennadius) from three host plants: cotton (Gossypium herbaceum), okra (Abelmoschus esculentus L.), and China rose (Hibiscus rosa-sinensis) belonging to the family Malvaceae. The presence of four secondary endosymbionts Rickettsia, Wolbachia, Cardinium, and Arsenophonus was checked in Bemisia tabaci populations. Phylogenetic analyses grounded on the mitochondrial cytochrome oxidase I gene (mtCO1) unveiled the presence of Asia 1, Asia II 1, and Asia II 7 genetic groups for Bemisia tabaci on abovementioned crops. Individuals were examined for symbiotic bacterial infection with specific primers amplifying the 16S rRNA gene for Portiera, Rickettsia, Cardinium, and Wolbachia, and the 23S rRNA gene for Arsenophonus. The results show that Portiera was present in all the Bemisia tabaci samples. However, variations were noted in the circulation frequencies of secondary endosymbionts among the Bemisia tabaci populations. A significant difference was noticed in the distribution frequency of Rickettsia between cotton and China rose or okra with their p values as 0.016 and 0.033 respectively. The uneven incidence of secondary endosymbionts ropes the assumption that each endosymbiotic bacterium not only has a role in the endurance but may contribute to the polyphagous nature of Bemisia tabaci. It also brings an uncomplicated evidence for progressive studies on control measures of this notorious insect pest.}, } @article {pmid30337547, year = {2018}, author = {Morioka, E and Oida, M and Tsuchida, T and Ikeda, M}, title = {Nighttime activities and peripheral clock oscillations depend on Wolbachia endosymbionts in flies.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {15432}, pmid = {30337547}, issn = {2045-2322}, support = {JP16H04651//Japan Society for the Promotion of Science (JSPS)/International ; JP16H04651//Japan Society for the Promotion of Science (JSPS)/International ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Circadian Clocks/genetics/*physiology ; Circadian Rhythm/genetics/*physiology ; Crosses, Genetic ; Drosophila Proteins/genetics ; Drosophila melanogaster/*microbiology/physiology ; Female ; Genes, Reporter ; *Host Microbial Interactions ; Locomotion/*physiology/radiation effects ; Male ; Period Circadian Proteins/genetics ; Photic Stimulation ; Symbiosis/physiology ; Tetracycline/pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {Wolbachia are ubiquitous bacterial endosymbionts of arthropods and affect host gene expression. Although Wolbachia infections were suggested to modulate sleep in flies, their influence on the circadian clock remained obscure. Here, we screened bacterial symbionts in a laboratory Drosophila melanogaster colony, and observed widespread infections of wMel strain Wolbachia. We established a Wolbachia-free strain from a clock gene reporter strain, period-luciferase (per-luc). Temperature (19-29 °C)-compensated free-running periods were detected regardless of infections which may reflect the lack of wMel infections in central circadian pacemaker neurons. However, locomotor activity levels during the night or subjective night were significantly amplified in uninfected flies. Moreover, the behavioral phenotype of F1 offspring of an uninfected female and infected male resembled that of uninfected flies. This trait is consistent with maternal transmission of Wolbachia infection. Interestingly, per-luc activities in headless bodies, as an index of peripheral circadian oscillators, were severely damped in uninfected flies. Additionally, circadian amplitudes of PER immunoreactivities in Malpighian tubules were reduced in uninfected flies. These results demonstrate that Wolbachia boost fly peripheral clock oscillations and diurnal behavioral patterns. Genetic mechanisms underlying behavioral rhythms have been widely analyzed using mutant flies whereas screening of Wolbachia will be necessary for future studies.}, } @article {pmid30321239, year = {2018}, author = {Bonneau, M and Landmann, F and Labbé, P and Justy, F and Weill, M and Sicard, M}, title = {The cellular phenotype of cytoplasmic incompatibility in Culex pipiens in the light of cidB diversity.}, journal = {PLoS pathogens}, volume = {14}, number = {10}, pages = {e1007364}, pmid = {30321239}, issn = {1553-7374}, mesh = {Animals ; Animals, Genetically Modified/genetics/*microbiology ; Bacterial Proteins/genetics/*metabolism ; Culex/genetics/*microbiology ; Cytoplasm/*microbiology/pathology ; Drosophila melanogaster/genetics/*microbiology ; Female ; Gram-Negative Bacterial Infections/microbiology/pathology ; Host Specificity ; Male ; Phenotype ; *Polymorphism, Genetic ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria, widespread among arthropods thanks to host reproductive manipulations that increase their prevalence into host populations. The most commonly observed manipulation is cytoplasmic incompatibility (CI). CI leads to embryonic death in crosses between i) infected males and uninfected females and ii) individuals infected with incompatible Wolbachia strains. CI can be conceptualized as a toxin-antidote system where a toxin deposited by Wolbachia in the sperm would induce embryonic death unless countered by an antidote produced by Wolbachia present in the eggs. In Drosophila melanogaster, transgenic expression of Wolbachia effector cidB revealed its function of CI-inducing toxin. Moreover in Culex pipiens, the diversity of cidB variants present in wPip strains accounts for the diversity in crossing-types. We conducted cytological analyses to determine the CI mechanisms that lead to embryonic death in C. pipiens, and assess whether diversity in crossing-types could be based on variations in these mechanisms. We revealed that paternal chromatin condensation and segregation defects during the first embryonic division are always responsible for embryonic death. The strongest observed defects lead to an exclusion of the paternal chromatin from the first zygotic division, resulting in haploid embryos unable to hatch. The proportion of unhatched haploid embryos, developing with only maternal chromatin, which reflects the frequency of strong defects can be considered as a proxy of CI intensity at the cellular level. We thus studied the putative effect of variations in crossing types and cidB diversification on CI defects intensity. Incompatible crosses involving distinct wPip strains revealed that CI defects intensity depends on the Wolbachia strains hosted by the males and is linked to the diversity of cidB genes harbored in their genomes. These results support that, additionally to its implication in C. pipiens crossing type variability, cidB diversification also influences the strength of CI embryonic defects.}, } @article {pmid30319574, year = {2018}, author = {Santos-Garcia, D and Juravel, K and Freilich, S and Zchori-Fein, E and Latorre, A and Moya, A and Morin, S and Silva, FJ}, title = {To B or Not to B: Comparative Genomics Suggests Arsenophonus as a Source of B Vitamins in Whiteflies.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {2254}, pmid = {30319574}, issn = {1664-302X}, abstract = {Insect lineages feeding on nutritionally restricted diets such as phloem sap, xylem sap, or blood, were able to diversify by acquiring bacterial species that complement lacking nutrients. These bacteria, considered obligate/primary endosymbionts, share a long evolutionary history with their hosts. In some cases, however, these endosymbionts are not able to fulfill all of their host's nutritional requirements, driving the acquisition of additional symbiotic species. Phloem-feeding members of the insect family Aleyrodidae (whiteflies) established an obligate relationship with Candidatus Portiera aleyrodidarum, which provides its hots with essential amino acids and carotenoids. In addition, many whitefly species harbor additional endosymbionts which may potentially further supplement their host's diet. To test this hypothesis, genomes of several endosymbionts of the whiteflies Aleurodicus dispersus, Aleurodicus floccissimus and Trialeurodes vaporariorum were analyzed. In addition to Portiera, all three species were found to harbor one Arsenophonus and one Wolbachia endosymbiont. A comparative analysis of Arsenophonus genomes revealed that although all three are capable of synthesizing B vitamins and cofactors, such as pyridoxal, riboflavin, or folate, their genomes and phylogenetic relationship vary greatly. Arsenophonus of A. floccissimus and T. vaporariorum belong to the same clade, and display characteristics of facultative endosymbionts, such as large genomes (3 Mb) with thousands of genes and pseudogenes, intermediate GC content, and mobile genetic elements. In contrast, Arsenophonus of A. dispersus belongs to a different lineage and displays the characteristics of a primary endosymbiont-a reduced genome (670 kb) with ~400 genes, 32% GC content, and no mobile genetic elements. However, the presence of 274 pseudogenes suggests that this symbiotic association is more recent than other reported primary endosymbionts of hemipterans. The gene repertoire of Arsenophonus of A. dispersus is completely integrated in the symbiotic consortia, and the biosynthesis of most vitamins occurs in shared pathways with its host. In addition, Wolbachia endosymbionts have also retained the ability to produce riboflavin, flavin adenine dinucleotide, and folate, and may make a nutritional contribution. Taken together, our results show that Arsenophonus hold a pivotal place in whitefly nutrition by their ability to produce B vitamins.}, } @article {pmid30314399, year = {2018}, author = {Paris, V and Cottingham, E and Ross, PA and Axford, JK and Hoffmann, AA}, title = {Effects of Alternative Blood Sources on Wolbachia Infected Aedes aegypti Females within and across Generations.}, journal = {Insects}, volume = {9}, number = {4}, pages = {}, pmid = {30314399}, issn = {2075-4450}, abstract = {Wolbachia bacteria have been identified as a tool for reducing the transmission of arboviruses transmitted by Aedes aegypti. Research groups around the world are now mass rearing Wolbachia-infected Ae. aegypti for deliberate release. We investigated the fitness impact of a crucial element of mass rearing: the blood meal required by female Ae. aegypti to lay eggs. Although Ae. aegypti almost exclusively feed on human blood, it is often difficult to use human blood in disease-endemic settings. When females were fed on sheep or pig blood rather than human blood, egg hatch rates decreased in all three lines tested (uninfected, or infected by wMel, or wAlbB Wolbachia). This finding was particularly pronounced when fed on sheep blood, although fecundity was not affected. Some of these effects persisted after an additional generation on human blood. Attempts to keep populations on sheep and pig blood sources only partly succeeded, suggesting that strong adaptation is required to develop a stably infected line on an alternative blood source. There was a decrease in Wolbachia density when Ae. aegypti were fed on non-human blood sources. Density increased in lines kept for multiple generations on the alternate sources but was still reduced relative to lines kept on human blood. These findings suggest that sheep and pig blood will entail a cost when used for maintaining Wolbachia-infected Ae. aegypti. These costs should be taken into account when planning mass release programs.}, } @article {pmid30311871, year = {2018}, author = {Taylor, MJ and Bordenstein, SR and Slatko, B}, title = {Microbe Profile: Wolbachia: a sex selector, a viral protector and a target to treat filarial nematodes.}, journal = {Microbiology (Reading, England)}, volume = {164}, number = {11}, pages = {1345-1347}, pmid = {30311871}, issn = {1465-2080}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R21 AI133522/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology ; Filarioidea/*microbiology ; Genome, Bacterial/genetics ; Host Specificity/*physiology ; Humans ; Symbiosis/physiology ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia is the most widespread genus of endosymbiotic bacteria in the animal world, infecting a diverse range of arthropods and nematodes. A broad spectrum of associations from parasitism to mutualism occur, with a tendency to drive reproductive manipulation or influence host fecundity to spread infection through host populations. These varied effects of Wolbachia are exploited for public health benefits. Notably, the protection of insect hosts from viruses is being tested as a potential control strategy for human arboviruses, and the mutualistic relationship with filarial nematodes makes Wolbachia a target for antibiotic therapy of human and veterinary nematode diseases.}, } @article {pmid30311675, year = {2018}, author = {Leftwich, PT and Hutchings, MI and Chapman, T}, title = {Diet, Gut Microbes and Host Mate Choice: Understanding the significance of microbiome effects on host mate choice requires a case by case evaluation.}, journal = {BioEssays : news and reviews in molecular, cellular and developmental biology}, volume = {40}, number = {12}, pages = {e1800053}, doi = {10.1002/bies.201800053}, pmid = {30311675}, issn = {1521-1878}, support = {BB/K000489/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Alleles ; Animals ; Biological Evolution ; *Diet ; Drosophila melanogaster/physiology ; Female ; Gastrointestinal Microbiome/*physiology ; Male ; Mating Preference, Animal/*physiology ; Symbiosis ; Wolbachia/physiology ; }, abstract = {All organisms live in close association with microbes. However, not all such associations are meaningful in an evolutionary context. Current debate concerns whether hosts and microbes are best described as communities of individuals or as holobionts (selective units of hosts plus their microbes). Recent reports that assortative mating of hosts by diet can be mediated by commensal gut microbes have attracted interest as a potential route to host reproductive isolation (RI). Here, the authors discuss logical problems with this line of argument. The authors briefly review how microbes can affect host mating preferences and evaluate recent findings from fruitflies. Endosymbionts can potentially influence host RI given stable and recurrent co-association of hosts and microbes over evolutionary time. However, observations of co-occurrence of microbes and hosts are ripe for misinterpretation and such associations will rarely represent a meaningful holobiont. A framework in which hosts and their microbes are independent evolutionary units provides the only satisfactory explanation for the observed range of effects and associations.}, } @article {pmid30311439, year = {2019}, author = {Zhu, YX and Song, YL and Hoffmann, AA and Jin, PY and Huo, SM and Hong, XY}, title = {A change in the bacterial community of spider mites decreases fecundity on multiple host plants.}, journal = {MicrobiologyOpen}, volume = {8}, number = {6}, pages = {e00743}, pmid = {30311439}, issn = {2045-8827}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Female ; Fertility ; Host Specificity ; Male ; *Microbiota ; Plants/parasitology ; Spiroplasma/classification/genetics/isolation & purification ; Tetranychidae/*microbiology/*physiology ; Wolbachia/classification/genetics/isolation & purification ; }, abstract = {Bacterial symbionts may influence the fitness of their herbivore hosts, but such effects have been poorly studied across most invertebrate groups. The spider mite, Tetranychus truncatus, is a polyphagous agricultural pest harboring various bacterial symbionts whose function is largely unknown. Here, by using a high-throughput 16S rRNA amplicon sequencing approach, we characterized the bacterial diversity and community composition of spider mites fed on five host plants after communities were modified following tetracycline exposure. We demonstrated that spider mite bacterial diversity and community composition were significantly affected by host plants and antibiotics. In particular, the abundance of the maternally inherited endosymbionts Wolbachia and Spiroplasma significantly differed among spider mites that were reared on different plant species and were completely removed by antibiotics. There was an overall tendency for daily fecundity to be lower in the mites with reduced bacterial diversity following the antibiotic treatment. Our data suggest that host plants and antibiotics can shape spider mite bacterial communities and that bacterial symbionts improve mite performance.}, } @article {pmid30305458, year = {2018}, author = {Bakovic, V and Schebeck, M and Telschow, A and Stauffer, C and Schuler, H}, title = {Correction to 'Spatial spread of Wolbachia in Rhagoletis cerasi populations'.}, journal = {Biology letters}, volume = {14}, number = {10}, pages = {}, doi = {10.1098/rsbl.2018.0683}, pmid = {30305458}, issn = {1744-957X}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, } @article {pmid30283652, year = {2017}, author = {Kageyama, D and Ohno, M and Sasaki, T and Yoshido, A and Konagaya, T and Jouraku, A and Kuwazaki, S and Kanamori, H and Katayose, Y and Narita, S and Miyata, M and Riegler, M and Sahara, K}, title = {Feminizing Wolbachia endosymbiont disrupts maternal sex chromosome inheritance in a butterfly species.}, journal = {Evolution letters}, volume = {1}, number = {5}, pages = {232-244}, pmid = {30283652}, issn = {2056-3744}, abstract = {Wolbachia is a maternally inherited ubiquitous endosymbiotic bacterium of arthropods that displays a diverse repertoire of host reproductive manipulations. For the first time, we demonstrate that Wolbachia manipulates sex chromosome inheritance in a sexually reproducing insect. Eurema mandarina butterfly females on Tanegashima Island, Japan, are infected with the wFem Wolbachia strain and produce all-female offspring, while antibiotic treatment results in male offspring. Fluorescence in situ hybridization (FISH) revealed that wFem-positive and wFem-negative females have Z0 and WZ sex chromosome sets, respectively, demonstrating the predicted absence of the W chromosome in wFem-infected lineages. Genomic quantitative polymerase chain reaction (qPCR) analysis showed that wFem-positive females lay only Z0 eggs that carry a paternal Z, whereas females from lineages that are naturally wFem-negative lay both WZ and ZZ eggs. In contrast, antibiotic treatment of adult wFem females resulted in the production of Z0 and ZZ eggs, suggesting that this Wolbachia strain can disrupt the maternal inheritance of Z chromosomes. Moreover, most male offspring produced by antibiotic-treated wFem females had a ZZ karyotype, implying reduced survival of Z0 individuals in the absence of feminizing effects of Wolbachia. Antibiotic treatment of wFem-infected larvae induced male-specific splicing of the doublesex (dsx) gene transcript, causing an intersex phenotype. Thus, the absence of the female-determining W chromosome in Z0 individuals is functionally compensated by Wolbachia-mediated conversion of sex determination. We discuss how Wolbachia may manipulate the host chromosome inheritance and that Wolbachia may have acquired this coordinated dual mode of reproductive manipulation first by the evolution of female-determining function and then cytoplasmically induced disruption of sex chromosome inheritance.}, } @article {pmid30275487, year = {2018}, author = {de Moraes, LA and Muller, C and Bueno, RCOF and Santos, A and Bello, VH and De Marchi, BR and Watanabe, LFM and Marubayashi, JM and Santos, BR and Yuki, VA and Takada, HM and de Barros, DR and Neves, CG and da Silva, FN and Gonçalves, MJ and Ghanim, M and Boykin, L and Pavan, MA and Krause-Sakate, R}, title = {Distribution and phylogenetics of whiteflies and their endosymbiont relationships after the Mediterranean species invasion in Brazil.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {14589}, pmid = {30275487}, issn = {2045-2322}, mesh = {Alphaproteobacteria/classification/genetics/*isolation & purification ; Animals ; Brazil ; Electron Transport Complex IV/genetics ; Gammaproteobacteria/classification/genetics/*isolation & purification ; Genetic Variation ; Hemiptera/*classification/genetics/*growth & development/microbiology ; *Introduced Species ; *Phylogeography ; *Symbiosis ; }, abstract = {The Bemisia tabaci is a polyphagous insect and a successful vector of plant viruses. B. tabaci is a species complex and in Brazil native species from the New World (NW) group, as well as the invasive species, Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) were reported. For better understanding the distribution of the different species four years after the Mediterranean species invasion in Brazil, whiteflies were collected from 237 locations throughout the country between the years of 2013 and 2017, species were identified and the facultative endosymbionts detected. The survey revealed that MEAM1 was the prevalent species found on major crops across Brazil. It is the only species present in North, Northwestern and Central Brazil and was associated with virus-infected plants. MED was found in five States from Southeast to South regions, infesting mainly ornamental plants and was not associated with virus-infected plants. The prevalent endosymbionts identified in MEAM1 were Hamiltonella and Rickettsia; and the mtCOI analysis revealed low genetic diversity for MEAM1. In contrast, several different endosymbionts were identified in MED including Hamiltonella, Rickettsia, Wolbachia and Arsenophonus; and two distinct genetic groups were found based on the mtCOI analysis. Monitoring the distribution of the whiteflies species in Brazil is essential for proper management of this pest.}, } @article {pmid30274467, year = {2018}, author = {van den Hurk, AF}, title = {From Incriminating Stegomyia fasciata to Releasing Wolbachia pipientis: Australian Research on the Dengue Virus Vector, Aedes aegypti, and Development of Novel Strategies for Its Surveillance and Control.}, journal = {Tropical medicine and infectious disease}, volume = {3}, number = {3}, pages = {}, pmid = {30274467}, issn = {2414-6366}, abstract = {Globally, the dengue viruses (DENVs) infect approximately 300 million people annually. Australia has a history of epidemic dengue, with outbreaks in the early decades of the twentieth century responsible for tens of thousands of cases. Seminal experiments conducted by Australian scientists during these outbreaks were the first to incriminate Aedes aegypti as a major vector of dengue viruses. One hundred years later, Australian scientists are playing a lead role in the development of surveillance and suppression strategies that target this mosquito species. Surveillance of Ae. aegypti populations and their associated dengue risk was greatly improved by understanding the contribution of key premises, key containers, and cryptic larval habitats to mosquito productivity, and, more recently, the development of novel adult traps. In terms of mosquito control, targeted indoor residual pyrethroid spraying and community-based biological control utilizing predatory copepods can significantly reduce Ae. aegypti populations. The release of Ae. aegypti transinfected with the virus-blocking bacterium, Wolbachia, provides a promising strategy for limiting DENV transmission. These diverse strategies developed by Australian scientists have the potential to alleviate the burden of dengue in the future, whether it is at the local level or as part of a country-wide program.}, } @article {pmid30266092, year = {2018}, author = {Pers, D and Lynch, JA}, title = {Ankyrin domain encoding genes from an ancient horizontal transfer are functionally integrated into Nasonia developmental gene regulatory networks.}, journal = {Genome biology}, volume = {19}, number = {1}, pages = {148}, pmid = {30266092}, issn = {1474-760X}, support = {R03 HD078578/HD/NICHD NIH HHS/United States ; R03 HD087476/HD/NICHD NIH HHS/United States ; 1R03HD087476/NH/NIH HHS/United States ; 1R03HD078578/NH/NIH HHS/United States ; }, mesh = {Animals ; *Ankyrin Repeat ; Body Patterning ; *Gene Expression Regulation, Developmental ; *Gene Regulatory Networks ; Gene Transfer, Horizontal ; Genes, Insect ; Insect Proteins/chemistry/*genetics/metabolism ; Wasps/embryology/*genetics ; }, abstract = {BACKGROUND: How regulatory networks incorporate additional components and how novel genes are functionally integrated into well-established developmental processes are two important and intertwined questions whose answers have major implications for understanding the evolution of development. We recently discovered a set of lineage-restricted genes with strong and specific expression patterns along the dorsal-ventral (DV) axis of the embryo of the wasp Nasonia that may serve as a powerful system for addressing these questions. We sought to both understand the evolutionary history of these genes and to determine their functions in the Nasonia DV patterning system.

RESULTS: We have found that the novel DV genes are part of a large family of rapidly duplicating and diverging ankyrin domain-encoding genes that originated most likely by horizontal transfer from a prokaryote in a common ancestor of the wasp superfamily Chalcidoidea. We tested the function of those ankyrin-encoding genes expressed along the DV axis and found that they participate in early embryonic DV patterning. We also developed a new wasp model system (Melittobia) and found that some functional integration of ankyrin genes have been preserved for over 90 million years.

CONCLUSIONS: Our results indicate that regulatory networks can incorporate novel genes that then become necessary for stable and repeatable outputs. Even a modest role in developmental networks may be enough to allow novel or duplicate genes to be maintained in the genome and become fully integrated network components.}, } @article {pmid30261054, year = {2018}, author = {Carpinone, EM and Li, Z and Mills, MK and Foltz, C and Brannon, ER and Carlow, CKS and Starai, VJ}, title = {Identification of putative effectors of the Type IV secretion system from the Wolbachia endosymbiont of Brugia malayi.}, journal = {PloS one}, volume = {13}, number = {9}, pages = {e0204736}, pmid = {30261054}, issn = {1932-6203}, support = {R01 AI100913/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Bacterial Proteins/biosynthesis/genetics ; Brugia malayi/*microbiology ; Gene Expression Regulation, Bacterial/*physiology ; Saccharomyces cerevisiae/genetics/metabolism ; *Symbiosis ; *Type IV Secretion Systems/genetics/metabolism ; *Wolbachia/genetics/metabolism ; }, abstract = {Wolbachia is an unculturable, intracellular bacterium that persists within an extremely broad range of arthropod and parasitic nematode hosts, where it is transmitted maternally to offspring via vertical transmission. In the filarial nematode Brugia malayi, a causative agent of human lymphatic filariasis, Wolbachia is an endosymbiont, and its presence is essential for proper nematode development, survival, and pathogenesis. While the elucidation of Wolbachia:nematode interactions that promote the bacterium's intracellular persistence is of great importance, research has been hampered due to the fact that Wolbachia cannot be cultured in the absence of host cells. The Wolbachia endosymbiont of B. malayi (wBm) has an active Type IV secretion system (T4SS). Here, we have screened 47 putative T4SS effector proteins of wBm for their ability to modulate growth or the cell biology of a typical eukaryotic cell, Saccharomyces cerevisiae. Five candidates strongly inhibited yeast growth upon expression, and 6 additional proteins showed toxicity in the presence of zinc and caffeine. Studies on the uptake of an endocytic vacuole-specific fluorescent marker, FM4-64, identified 4 proteins (wBm0076 wBm00114, wBm0447 and wBm0152) involved in vacuole membrane dynamics. The WAS(p)-family protein, wBm0076, was found to colocalize with yeast cortical actin patches and disrupted actin cytoskeleton dynamics upon expression. Deletion of the Arp2/3-activating protein, Abp1p, provided resistance to wBm0076 expression, suggesting a role for wBm0076 in regulating eukaryotic actin dynamics and cortical actin patch formation. Furthermore, wBm0152 was found to strongly disrupt endosome:vacuole cargo trafficking in yeast. This study provides molecular insight into the potential role of the T4SS in the Wolbachia endosymbiont:nematode relationship.}, } @article {pmid30258200, year = {2018}, author = {Kaczmarczyk, A and Kucharczyk, H and Kucharczyk, M and Kapusta, P and Sell, J and Zielińska, S}, title = {First insight into microbiome profile of fungivorous thrips Hoplothrips carpathicus (Insecta: Thysanoptera) at different developmental stages: molecular evidence of Wolbachia endosymbiosis.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {14376}, pmid = {30258200}, issn = {2045-2322}, mesh = {Actinobacteria/genetics/isolation & purification/physiology ; Alphaproteobacteria/genetics/isolation & purification/physiology ; Animals ; Bacteria/genetics/isolation & purification ; Firmicutes/genetics/isolation & purification/physiology ; *Microbiota ; Phylogeny ; Proteobacteria/genetics/isolation & purification/physiology ; *Symbiosis ; Thysanoptera/*microbiology/*physiology ; Wolbachia/genetics/*isolation & purification/*physiology ; }, abstract = {Insects' exoskeleton, gut, hemocoel, and cells are colonized by various microorganisms that often play important roles in their host life. Moreover, insects are frequently infected by vertically transmitted symbionts that can manipulate their reproduction. The aims of this study were the characterization of bacterial communities of four developmental stages of the fungivorous species Hoplothrips carpathicus (Thysanoptera: Phlaeothripidae), verification of the presence of Wolbachia, in silico prediction of metabolic potentials of the microorganisms, and sequencing its mitochondrial COI barcode. Taxonomy-based analysis indicated that the bacterial community of H. carpathicus contained 21 bacterial phyla. The most abundant phyla were Proteobacteria, Actinobacteria, Bacterioidetes and Firmicutes, and the most abundant classes were Alphaproteobacteria, Actinobacteria, Gammaproteobacteria and Betaproteobacteria, with different proportions in the total share. For pupa and imago (adult) the most abundant genus was Wolbachia, which comprised 69.95% and 56.11% of total bacterial population respectively. Moreover, similarity analysis of bacterial communities showed that changes in microbiome composition are congruent with the successive stages of H. carpathicus development. PICRUSt analysis predicted that each bacterial community should be rich in genes involved in membrane transport, amino acid metabolism, carbohydrate metabolism, replication and repair processes.}, } @article {pmid30250462, year = {2018}, author = {Hegde, S and Khanipov, K and Albayrak, L and Golovko, G and Pimenova, M and Saldaña, MA and Rojas, MM and Hornett, EA and Motl, GC and Fredregill, CL and Dennett, JA and Debboun, M and Fofanov, Y and Hughes, GL}, title = {Microbiome Interaction Networks and Community Structure From Laboratory-Reared and Field-Collected Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus Mosquito Vectors.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {2160}, pmid = {30250462}, issn = {1664-302X}, support = {U01CK000512/ACL/ACL HHS/United States ; U01 CK000512/CK/NCEZID CDC HHS/United States ; R21 AI124452/AI/NIAID NIH HHS/United States ; T32 AI007526/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; }, abstract = {Microbial interactions are an underappreciated force in shaping insect microbiome communities. Although pairwise patterns of symbiont interactions have been identified, we have a poor understanding regarding the scale and the nature of co-occurrence and co-exclusion interactions within the microbiome. To characterize these patterns in mosquitoes, we sequenced the bacterial microbiome of Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus caught in the field or reared in the laboratory and used these data to generate interaction networks. For collections, we used traps that attracted host-seeking or ovipositing female mosquitoes to determine how physiological state affects the microbiome under field conditions. Interestingly, we saw few differences in species richness or microbiome community structure in mosquitoes caught in either trap. Co-occurrence and co-exclusion analysis identified 116 pairwise interactions substantially increasing the list of bacterial interactions observed in mosquitoes. Networks generated from the microbiome of Ae. aegypti often included highly interconnected hub bacteria. There were several instances where co-occurring bacteria co-excluded a third taxa, suggesting the existence of tripartite relationships. Several associations were observed in multiple species or in field and laboratory-reared mosquitoes indicating these associations are robust and not influenced by environmental or host factors. To demonstrate that microbial interactions can influence colonization of the host, we administered symbionts to Ae. aegypti larvae that either possessed or lacked their resident microbiota. We found that the presence of resident microbiota can inhibit colonization of particular bacterial taxa. Our results highlight that microbial interactions in mosquitoes are complex and influence microbiome composition.}, } @article {pmid30247733, year = {2018}, author = {Ritchie, SA}, title = {Wolbachia and the near cessation of dengue outbreaks in Northern Australia despite continued dengue importations via travellers.}, journal = {Journal of travel medicine}, volume = {25}, number = {1}, pages = {}, doi = {10.1093/jtm/tay084}, pmid = {30247733}, issn = {1708-8305}, mesh = {Australia ; Dengue/*prevention & control/transmission ; Disease Outbreaks/*prevention & control ; Humans ; Mosquito Control/*trends ; Queensland/epidemiology ; *Wolbachia ; }, } @article {pmid30244151, year = {2019}, author = {Schön, I and Kamiya, T and Van den Berghe, T and Van den Broecke, L and Martens, K}, title = {Novel Cardinium strains in non-marine ostracod (Crustacea) hosts from natural populations.}, journal = {Molecular phylogenetics and evolution}, volume = {130}, number = {}, pages = {406-415}, doi = {10.1016/j.ympev.2018.09.008}, pmid = {30244151}, issn = {1095-9513}, mesh = {Animals ; Aquatic Organisms/*microbiology ; Bacteroidetes/genetics/*physiology ; Base Sequence ; Crustacea/genetics/*microbiology ; Diptera/genetics ; Electron Transport Complex IV/genetics ; Genetic Variation ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; Symbiosis ; }, abstract = {Endosymbiotic bacteria are known from many metazoan taxa, where they manipulate host biology and reproduction. Here, we used classic PCR amplification and direct DNA sequencing with universal primers for four different endosymbionts to test for their presence in more than 300 specimens of three recent non-marine ostracod superfamilies from different geographic areas and aquatic habitats. We verified these results with "high throughput" amplicon sequencing of 16S of nine selected specimens and evolutionary placement algorithms. The phylogenetic position of endosymbionts detected in ostracod hosts was compared to known endosymbionts from other metazoans. While Wolbachia, Spiroplasma and Rickettsia are absent, we find evidence for the general presence of Cardinium bacteria in natural populations of various non-marine ostracod species. Phylogenetic reconstructions based on Cardinium 16S data and estimates of genetic distances both indicate that Cardinium from ostracods are distantly related to Cardinium from Diptera and Nematoda but represent novel strains with a monophyletic origin. Cardinium bacteria from different ostracod hosts have genetic distances of up to 3.8%, providing evidence against recent and frequent horizontal transmissions amongst the three ostracod superfamilies. High throughput sequencing reveals more than 400 different 16S amplicon sequence variants in the investigated ostracods as well as the presence of different Cardinium strains within individual Eucypris virens and Heterocypris hosts. These results call for future, more in-depth investigations. Mapping Cardinium infections on COI trees of non-marine ostracod hosts shows that the occurrence of these endosymbionts is not linked to genetic species identity or phylogenetic host groups and, except for one ostracod morphospecies, prevalence never reaches 100%.}, } @article {pmid30239661, year = {2018}, author = {Guégan, M and Minard, G and Tran, FH and Tran Van, V and Dubost, A and Valiente Moro, C}, title = {Short-term impacts of anthropogenic stressors on Aedes albopictus mosquito vector microbiota.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {12}, pages = {}, doi = {10.1093/femsec/fiy188}, pmid = {30239661}, issn = {1574-6941}, mesh = {Aedes/*microbiology ; Animals ; Anti-Bacterial Agents/*pharmacology ; Bacteroidetes/growth & development ; Chryseobacterium/growth & development ; Climate ; Dysbiosis/*chemically induced ; Flavobacteriaceae/growth & development ; Humans ; Microbiota/*drug effects ; Mosquito Vectors/microbiology ; Temperature ; Wolbachia/growth & development ; }, abstract = {Recent studies have highlighted the potential role of microbiota in the biology of the Aedes albopictus mosquito vector. This species is highly anthropogenic and exhibits marked ecological plasticity, with a resulting high potential to colonize a wide range of habitats-including anthropized areas-under various climatic conditions. We put forward the hypothesis that climate and anthropogenic activities, such as the use of antibiotics in agriculture and human medicine, might affect the mosquito-associated bacterial community. We thus studied the additive impact of a temperature decrease and antibiotic ingestion on the temporal dynamics of Ae. albopictus survival and its associated bacterial communities. The results showed no effects of disturbances on mosquito survival. However, short-term temperature impacts on bacterial diversity were observed, while both the community structure and bacterial diversity were affected by early antibiotic ingestion. The genera Elizabethkingia, Chryseobacterium and Wolbachia, as well as an unclassified member of the Bacteroidales order were particularly affected. Antibiotics negatively impacted Elizabethkingia abundance, while Chryseobacterium was completely eliminated following both disturbances, to the benefit of Wolbachia and the unclassified Bacteroidales species. These results generated fresh insight into the effects of climate and anthropogenic activities such as the use of antibiotics on mosquito microbiota.}, } @article {pmid30233514, year = {2018}, author = {Guo, Y and Hoffmann, AA and Xu, XQ and Mo, PW and Huang, HJ and Gong, JT and Ju, JF and Hong, XY}, title = {Vertical Transmission of Wolbachia Is Associated With Host Vitellogenin in Laodelphax striatellus.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {2016}, pmid = {30233514}, issn = {1664-302X}, abstract = {Wolbachia in host germ lines are essential for their vertical transmission to the next generation. It is unclear how the regulation of host oocyte development influences Wolbachia location and the mechanistic basis of transmission. Here, we investigated whether vitellogenin influences Wolbachia transmission in Laodelphax striatellus. Wolbachia increased in density and spread from the anterior tropharium to developing oocytes as ovaries developed. Microscopic observations indicated that Wolbachia invaded ovarioles from the tropharium of its anterior side rather than the pedicel side. Wolbachia utilized the host Vg transovarial transportation system to enter the ovaries and were transmitted from the tropharium into the developing oocytes through nutritive cords. These observations were supported by knocking down the Vg transcript, in which low Wolbachia titers were detected in ovaries and fewer Wolbachia were transmitted into oocytes. Our findings establish a link between the Vg-related mode of transovarial transmission and efficient maternal transmission of Wolbachia.}, } @article {pmid30233221, year = {2018}, author = {Cheng, YH and Lin, YJ and Chen, SC and You, SH and Chen, WY and Hsieh, NH and Yang, YF and Liao, CM}, title = {Assessing health burden risk and control effect on dengue fever infection in the southern region of Taiwan.}, journal = {Infection and drug resistance}, volume = {11}, number = {}, pages = {1423-1435}, pmid = {30233221}, issn = {1178-6973}, abstract = {BACKGROUND: The high prevalence of dengue in Taiwan and the consecutive large dengue outbreaks in the period 2014-2015 suggest that current control interventions are suboptimal. Understanding the effect of control effort is crucial to inform future control strategies.

OBJECTIVES: We developed a framework to measure season-based health burden risk from 2001 to 2014. We reconstructed various intervention coverage to assess the attributable effect of dengue infection control efforts.

MATERIALS AND METHODS: A dengue-mosquito-human transmission dynamic was used to quantify the vector-host interactions and to estimate the disease epidemics. We used disability-adjusted life years (DALYs) to assess health burden risk. A temperature-basic reproduction number (R0)-DALYs relationship was constructed to examine the potential impacts of temperature on health burden. Finally, a health burden risk model linked a control measure model to evaluate the effect of dengue control interventions.

RESULTS: We showed that R0 and DALYs peaked at 25°C with estimates of 2.37 and 1387, respectively. Results indicated that most dengue cases occurred in fall with estimated DALYs of 323 (267-379, 95% CI) at 50% risk probability. We found that repellent spray had by far the largest control effect with an effectiveness of ~71% in all seasons. Pesticide spray and container clean-up have both made important contributions to reducing prevalence/incidence. Repellent, pesticide spray, container clean-up together with Wolbachia infection suppress dengue outbreak by ~90%.

CONCLUSION: Our presented modeling framework provides a useful tool to measure dengue health burden risk and to quantify the effect of dengue control on dengue infection prevalence and disease incidence in the southern region of Taiwan.}, } @article {pmid30226138, year = {2018}, author = {Indriani, C and Ahmad, RA and Wiratama, BS and Arguni, E and Supriyati, E and Sasmono, RT and Kisworini, FY and Ryan, PA and O'Neill, SL and Simmons, CP and Utarini, A and Anders, KL}, title = {Baseline Characterization of Dengue Epidemiology in Yogyakarta City, Indonesia, before a Randomized Controlled Trial of Wolbachia for Arboviral Disease Control.}, journal = {The American journal of tropical medicine and hygiene}, volume = {99}, number = {5}, pages = {1299-1307}, pmid = {30226138}, issn = {1476-1645}, mesh = {Aedes/microbiology/*virology ; Animals ; Antibodies, Viral/blood ; Child ; Child, Preschool ; Dengue/*epidemiology/immunology ; Dengue Virus ; Hospitalization/statistics & numerical data ; Humans ; Incidence ; Indonesia/epidemiology ; Infant ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/virology ; Pest Control, Biological/*methods ; Prospective Studies ; Randomized Controlled Trials as Topic ; Seroepidemiologic Studies ; Surveys and Questionnaires ; Wolbachia/*physiology ; }, abstract = {Dengue is endemic in Indonesia. Here, we describe the epidemiology of dengue in the city of Yogyakarta, Central Java, as a prelude to implementation of a cluster-randomized trial of Wolbachia for the biocontrol of arboviral transmission. Surveillance records from 2006 to 2016 demonstrate seasonal oscillations of dengue incidence with varying magnitude. Two lines of evidence demonstrate a high force of infection; the hospitalized case burden of patients diagnosed with dengue hemorrhagic fever or dengue shock syndrome over the last decade consisted predominantly of children/adolescents, and a serosurvey of 314 healthy children aged 1-10 years found 68% possessed dengue virus-neutralizing antibodies. Finally, a mobility survey indicated children aged 1-10 years, and particularly 1-5 year-olds, spent most of their daytime hours at home. These findings inform the design of clinical trials to measure the impact of novel vector control methods such as Wolbachia introgression into Aedes aegypti mosquitoes, by providing baseline data on disease incidence and identifying subpopulations for recruitment into prospective studies of dengue virus infection and disease. The mobility survey findings indicate that in cluster trials of interventions applied at the community level, young children can reasonably be expected to spend most of their exposure time, in epidemiological terms, within the treatment arm to which they were randomized.}, } @article {pmid30219893, year = {2018}, author = {Zélé, F and Santos, JL and Godinho, DP and Magalhães, S}, title = {Wolbachia both aids and hampers the performance of spider mites on different host plants.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {12}, pages = {}, doi = {10.1093/femsec/fiy187}, pmid = {30219893}, issn = {1574-6941}, mesh = {Animals ; Bacteroidetes/metabolism ; Fabaceae/microbiology/parasitology ; Female ; Host Specificity ; Ipomoea nil/*microbiology/*parasitology ; Solanum lycopersicum/*microbiology/*parasitology ; Rickettsia/metabolism ; Solanum melongena/*microbiology/*parasitology ; Symbiosis/physiology ; Tetranychidae/metabolism/*microbiology ; Wolbachia/*metabolism ; }, abstract = {In the last few decades, many studies have revealed the potential role of arthropod bacterial endosymbionts in shaping the host range of generalist herbivores and their performance on different host plants, which, in turn, might affect endosymbiont distribution in herbivore populations. We tested this by measuring the prevalence of endosymbionts in natural populations of the generalist spider mite Tetranychus urticae on different host plants. Focusing on Wolbachia, we then analysed how symbionts affected mite life-history traits on the same host plants in the laboratory. Overall, the prevalences of Cardinium and Rickettsia were low, whereas that of Wolbachia was high, with the highest values on bean and eggplant and the lowest on morning glory, tomato and zuchini. Although most mite life-history traits were affected by the plant species only, Wolbachia infection was detrimental for the egg-hatching rate on morning glory and zucchini, and led to a more female-biased sex ratio on morning glory and eggplant. These results suggest that endosymbionts may affect the host range of polyphagous herbivores, both by aiding and hampering their performance, depending on the host plant and on the life-history trait that affects performance the most. Conversely, endosymbiont spread may be facilitated or hindered by the plants on which infected herbivores occur.}, } @article {pmid30202647, year = {2018}, author = {Pascar, J and Chandler, CH}, title = {A bioinformatics approach to identifying Wolbachia infections in arthropods.}, journal = {PeerJ}, volume = {6}, number = {}, pages = {e5486}, pmid = {30202647}, issn = {2167-8359}, abstract = {Wolbachia is the most widespread endosymbiont, infecting >20% of arthropod species, and capable of drastically manipulating the host's reproductive mechanisms. Conventionally, diagnosis has relied on PCR amplification; however, PCR is not always a reliable diagnostic technique due to primer specificity, strain diversity, degree of infection and/or tissue sampled. Here, we look for evidence of Wolbachia infection across a wide array of arthropod species using a bioinformatic approach to detect the Wolbachia genes ftsZ, wsp, and the groE operon in next-generation sequencing samples available through the NCBI Sequence Read Archive. For samples showing signs of infection, we attempted to assemble entire Wolbachia genomes, and in order to better understand the relationships between hosts and symbionts, phylogenies were constructed using the assembled gene sequences. Out of the 34 species with positively identified infections, eight species of arthropod had not previously been recorded to harbor Wolbachia infection. All putative infections cluster with known representative strains belonging to supergroup A or B, which are known to only infect arthropods. This study presents an efficient bioinformatic approach for post-sequencing diagnosis and analysis of Wolbachia infection in arthropods.}, } @article {pmid30196669, year = {2018}, author = {Liu, Y and He, B and Li, F and Li, K and Zhang, L and Li, X and Zhao, L}, title = {Molecular Identification of Bartonella melophagi and Wolbachia Supergroup F from Sheep Keds in Xinjiang, China.}, journal = {The Korean journal of parasitology}, volume = {56}, number = {4}, pages = {365-370}, pmid = {30196669}, issn = {1738-0006}, support = {31460655//National Natural Science Foundation of China/ ; HS201501//Tarim Animal Husbandry Science and Technology, Xinjiang Production & Construction Corps/ ; HS201801//Tarim Animal Husbandry Science and Technology, Xinjiang Production & Construction Corps/ ; }, mesh = {Animals ; Bacterial Proteins/*genetics ; Bartonella/classification/*genetics/*isolation & purification/pathogenicity ; China ; DNA, Bacterial/genetics ; Diptera/*microbiology ; Phylogeny ; Polymerase Chain Reaction/methods ; RNA, Bacterial/*genetics ; RNA, Ribosomal, 16S/*genetics ; Sequence Analysis, DNA ; Sheep/parasitology ; Wolbachia/classification/*genetics/*isolation & purification/pathogenicity ; }, abstract = {To confirm that Bartonella and Wolbachia were carried by sheep keds (Melophagus ovinus) in southern Xinjiang of China, 17 M. ovinus samples, which were collected in Aksu Prefecture, Xinjiang, were randomly selected. In this study, the Bartonella gltA and Wolbachia 16S rRNA gene were amplified through conventional PCR and the sequence of those amplified products, were analyzed. The results demonstrated that Bartonella was carried by all of the 17 sheep keds and Wolbachia was carried by 15 out of them. Bartonella was identified as B. melophagi. Three strains of Wolbachia were supergroup F and 1 strain has not been confirmed yet. It is the first report about Wolbachia supergroup F was found in sheep keds and provided the molecular evidence that B. melophagi and Wolbachia supergroup F were carried by sheep keds in Aksu Prefecture of southern Xinjiang, China. The 2 pathogens were found in sheep keds around Taklimakan Desert for the first time.}, } @article {pmid30193778, year = {2018}, author = {Lopez, V and Cortesero, AM and Poinsot, D}, title = {Influence of the symbiont Wolbachia on life history traits of the cabbage root fly (Delia radicum).}, journal = {Journal of invertebrate pathology}, volume = {158}, number = {}, pages = {24-31}, doi = {10.1016/j.jip.2018.09.002}, pmid = {30193778}, issn = {1096-0805}, mesh = {Animals ; Diptera/*microbiology ; Female ; *Life History Traits ; Male ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia is an endocellular bacteria infecting arthropods and nematodes and is only transmitted vertically by females via the cytoplasm of the egg. It is often a manipulator of host reproduction, causing cytoplasmic incompatibility, thelytokous parthenogenesis, feminization or male killing, which all increase the proportion of infected females in the population. However, Wolbachia can modify life history traits of the host without causing the above phenotypes and each species illustrates the variability of relationships between this remarkably versatile symbiont and its many hosts. We have measured maternal transmission and the impact of a natural Wolbachia infection in the cabbage root fly Delia radicum, a major agricultural pest. We used a population that is polymorphic for the infection to ensure similar genetic and microbiome backgrounds between groups. Maternal transmission of the infection was 100% in our sample. We found no evidence of cytoplasmic incompatibility, thelytokous parthenogenesis, feminization nor male killing. Wolbachia infection significantly reduced hatch rate in infected eggs (by 10%) but improved larvo-nymphal viability sufficiently so that infected eggs nevertheless yielded as many adults as uninfected ones, albeit with a 1.5% longer total development time. Starved and infected ovipositing females suffered significantly reduced viability (20% higher mortality during a 3-day oviposition period) than uninfected females, but mortality was not higher in starved virgin females nor in starved males, suggesting that the energetic cost of the infection is only revealed in extreme conditions. Wolbachia had no effect on egg hatch time or offspring size. The apparently 100% vertical transmission and the significant but mutually compensating effects found suggest that infection might be nearly benign in this host and might only drift slowly, which would explain why the infection rate has been stable in our laboratory (approximately 50% individuals infected) for at least 30 generations.}, } @article {pmid30190541, year = {2018}, author = {Chung, M and Teigen, L and Liu, H and Libro, S and Shetty, A and Kumar, N and Zhao, X and Bromley, RE and Tallon, LJ and Sadzewicz, L and Fraser, CM and Rasko, DA and Filler, SG and Foster, JM and Michalski, ML and Bruno, VM and Dunning Hotopp, JC}, title = {Targeted enrichment outperforms other enrichment techniques and enables more multi-species RNA-Seq analyses.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {13377}, pmid = {30190541}, issn = {2045-2322}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Aspergillus fumigatus/*genetics ; Brugia malayi/*genetics ; *RNA, Bacterial/chemistry/genetics/isolation & purification ; *RNA, Fungal/chemistry/genetics/isolation & purification ; *RNA, Helminth/chemistry/genetics/isolation & purification ; *RNA, Messenger/chemistry/genetics/isolation & purification ; Sequence Analysis, RNA/*methods ; Wolbachia/*genetics ; }, abstract = {Enrichment methodologies enable the analysis of minor members in multi-species transcriptomic data. We compared the standard enrichment of bacterial and eukaryotic mRNA to a targeted enrichment using an Agilent SureSelect (AgSS) capture for Brugia malayi, Aspergillus fumigatus, and the Wolbachia endosymbiont of B. malayi (wBm). Without introducing significant systematic bias, the AgSS quantitatively enriched samples, resulting in more reads mapping to the target organism. The AgSS-enriched libraries consistently had a positive linear correlation with their unenriched counterparts (r[2] = 0.559-0.867). Up to a 2,242-fold enrichment of RNA from the target organism was obtained following a power law (r[2] = 0.90), with the greatest fold enrichment achieved in samples with the largest ratio difference between the major and minor members. While using a single total library for prokaryote and eukaryote enrichment from a single RNA sample could be beneficial for samples where RNA is limiting, we observed a decrease in reads mapping to protein coding genes and an increase in multi-mapping reads to rRNAs in AgSS enrichments from eukaryotic total RNA libraries compared to eukaryotic poly(A)-enriched libraries. Our results support a recommendation of using AgSS targeted enrichment on poly(A)-enriched libraries for eukaryotic captures, and total RNA libraries for prokaryotic captures, to increase the robustness of multi-species transcriptomic studies.}, } @article {pmid30190331, year = {2018}, author = {Leftwich, PT and Edgington, MP and Harvey-Samuel, T and Carabajal Paladino, LZ and Norman, VC and Alphey, L}, title = {Recent advances in threshold-dependent gene drives for mosquitoes.}, journal = {Biochemical Society transactions}, volume = {46}, number = {5}, pages = {1203-1212}, pmid = {30190331}, issn = {1470-8752}, support = {BBS/E/I/00007033/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BBS/E/I/00007038/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 110117/Z/15/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Chikungunya Fever ; Dengue ; Drosophila/microbiology ; *Gene Drive Technology ; Haploinsufficiency ; Heterozygote ; Malaria ; Models, Theoretical ; Mosquito Vectors/genetics/*physiology ; RNA Interference ; Reproduction ; Wolbachia ; }, abstract = {Mosquito-borne diseases, such as malaria, dengue and chikungunya, cause morbidity and mortality around the world. Recent advances in gene drives have produced control methods that could theoretically modify all populations of a disease vector, from a single release, making whole species less able to transmit pathogens. This ability has caused both excitement, at the prospect of global eradication of mosquito-borne diseases, and concern around safeguards. Drive mechanisms that require individuals to be released at high frequency before genes will spread can therefore be desirable as they are potentially localised and reversible. These include underdominance-based strategies and use of the reproductive parasite Wolbachia Here, we review recent advances in practical applications and mathematical analyses of these threshold-dependent gene drives with a focus on implementation in Aedes aegypti, highlighting their mechanisms and the role of fitness costs on introduction frequencies. Drawing on the parallels between these systems offers useful insights into practical, controlled application of localised drives, and allows us to assess the requirements needed for gene drive reversal.}, } @article {pmid30188519, year = {2018}, author = {Gilbert, JA and Melton, L}, title = {Verily project releases millions of factory-reared mosquitoes.}, journal = {Nature biotechnology}, volume = {36}, number = {9}, pages = {781-782}, pmid = {30188519}, issn = {1546-1696}, mesh = {Aedes/*microbiology ; Animals ; California ; Dengue/prevention & control ; Female ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology ; Wolbachia ; }, } @article {pmid30175240, year = {2018}, author = {Jeffries, CL and Rogers, ME and Walker, T}, title = {Establishment of a method for Lutzomyia longipalpis sand fly embryo microinjection: The first step towards potential novel control strategies for leishmaniasis.}, journal = {Wellcome open research}, volume = {3}, number = {}, pages = {55}, pmid = {30175240}, issn = {2398-502X}, support = {//Wellcome Trust/United Kingdom ; BB/H022406/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, abstract = {Leishmaniasis is a vector-borne parasitic disease transmitted by sand flies that affects 1.3 million people across 98 countries, with limited control strategies due to the lack of an available vaccine and the emergence of insecticide resistance. Novel control strategies that are being explored for mosquito-borne diseases, such as Wolbachia bacterial inhibition of pathogens and genetically modified insects (e.g. using CRISPR-Cas9 editing), rely on the ability to consistently inject embryos of the target species. Here we present a novel method to obtain and inject preblastoderm sand fly embryos of the genus Lutzomyia (Lu.)longipalpis, the principle vector of zoonotic visceral leishmaniasis in South America. The procedures required to obtain sufficiently young Lu. longipalpis colony embryos are described alongside a microinjection technique that permits rapid injection and minimal handling of small sand fly embryos post-injection. Using a strain of Wolbachia as a 'marker' for successful injection, our protocol produced early generation Wolbachia transinfected Lu. longipalpis lines, demonstrating its potential as the first step for use in novel applied strategies for sand fly control.}, } @article {pmid30165658, year = {2018}, author = {Cevidanes, A and Di Cataldo, S and Vera, F and Lillo, P and Millán, J}, title = {Molecular Detection of Vector-Borne Pathogens in Rural Dogs and Associated Ctenocephalides felis Fleas (Siphonaptera: Pulicidae) in Easter Island (Chile).}, journal = {Journal of medical entomology}, volume = {55}, number = {6}, pages = {1659-1663}, doi = {10.1093/jme/tjy141}, pmid = {30165658}, issn = {1938-2928}, mesh = {Animals ; Ctenocephalides/*microbiology ; Dogs/*parasitology ; Female ; Insect Vectors/*microbiology ; Male ; Polynesia ; }, abstract = {The presence of vector-borne pathogens of veterinary and public health interest have received little attention in Chile. In Easter Island, in particular, a Chilean territory in the southeastern Pacific Ocean, no information is available. To fill this gap, 153 rural dogs were inspected for ectoparasites during a sterilization campaign carried out in 2016. Fleas were observed in 46% of the dogs, and Ctenocephalides felis (Bouché, 1835) was the only species present. Morphological identification of fleas was genetically confirmed using conventional polymerase chain reaction targeting the cox2 gene. No tick was observed in any dog. The presence of DNA of Rickettsia sp. (gltA and ompA fragment genes), Anaplasmataceae (16S rRNA), and Bartonella sp. (16S-23S ribosomal RNA intergenic spacer) was investigated in blood samples of 70 of the dogs and in 126 fleas analyzed in 68 pools that included 1-5 fleas. Rickettsial DNA was detected in 97% (n = 66) of the flea pools. Of these, 57 showed between 99 and 100% identity for both genes with published sequences of Candidatus Rickettsia asemboensis (CRa), six with Rickettsia felis, and one with Candidatus Rickettsia senegalensis. For two pools, gltA amplicons were identical to CRa but ompB amplicions showed 99-100% identity with R. felis. Anaplasmataceae DNA was detected in 16% (n = 11) pools. Sequenced amplicons showed highest identity with the endosymbiont Wolbachia pipientis. Bartonella DNA, showing 99% identity to Bartonella clarridgeiae, was detected in one pool (1.4%). No positive reaction was observed for any dog. This is the first detection of members of the 'R. felis-like' group other than R. felis in Chile.}, } @article {pmid30161131, year = {2018}, author = {Bhadra, S and Riedel, TE and Saldaña, MA and Hegde, S and Pederson, N and Hughes, GL and Ellington, AD}, title = {Direct nucleic acid analysis of mosquitoes for high fidelity species identification and detection of Wolbachia using a cellphone.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {8}, pages = {e0006671}, pmid = {30161131}, issn = {1935-2735}, support = {R21 AI124452/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; R43 AI131948/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; U01 CK000512/CK/NCEZID CDC HHS/United States ; }, mesh = {Aedes/*genetics ; Animals ; Bacterial Proteins/genetics ; *Cell Phone ; Electron Transport Complex IV/genetics ; Nucleic Acid Amplification Techniques/*methods ; Software ; Wolbachia/*isolation & purification ; }, abstract = {Manipulation of natural mosquito populations using the endosymbiotic bacteria Wolbachia is being investigated as a novel strategy to reduce the burden of mosquito-borne viruses. To evaluate the efficacy of these interventions, it will be critical to determine Wolbachia infection frequencies in Aedes aegypti mosquito populations. However, current diagnostic tools are not well-suited to fit this need. Morphological methods cannot identify Wolbachia, immunoassays often suffer from low sensitivity and poor throughput, while PCR and spectroscopy require complex instruments and technical expertise, which restrict their use to centralized laboratories. To address this unmet need, we have used loop-mediated isothermal amplification (LAMP) and oligonucleotide strand displacement (OSD) probes to create a one-pot sample-to-answer nucleic acid diagnostic platform for vector and symbiont surveillance. LAMP-OSD assays can directly amplify target nucleic acids from macerated mosquitoes without requiring nucleic acid purification and yield specific single endpoint yes/no fluorescence signals that are observable to eye or by cellphone camera. We demonstrate cellphone-imaged LAMP-OSD tests for two targets, the Aedes aegypti cytochrome oxidase I (coi) gene and the Wolbachia surface protein (wsp) gene, and show a limit of detection of 4 and 40 target DNA copies, respectively. In a blinded test of 90 field-caught mosquitoes, the coi LAMP-OSD assay demonstrated 98% specificity and 97% sensitivity in identifying Ae. aegypti mosquitoes even after 3 weeks of storage without desiccant at 37°C. Similarly, the wsp LAMP-OSD assay readily identified the wAlbB Wolbachia strain in field-collected Aedes albopictus mosquitoes without generating any false positive signals. Modest technology requirements, minimal execution steps, simple binary readout, and robust accuracy make the LAMP-OSD-to-cellphone assay platform well suited for field vector surveillance in austere or resource-limited conditions.}, } @article {pmid30157257, year = {2018}, author = {Wenzel, MA and Douglas, A and Piertney, SB}, title = {Microbiome composition within a sympatric species complex of intertidal isopods (Jaera albifrons).}, journal = {PloS one}, volume = {13}, number = {8}, pages = {e0202212}, pmid = {30157257}, issn = {1932-6203}, mesh = {Animals ; Bacteria/classification/genetics/isolation & purification ; Bacteroidetes/genetics/isolation & purification ; Ecosystem ; Female ; Genetic Speciation ; Genetic Variation ; Host Microbial Interactions/genetics ; Isopoda/*microbiology ; Male ; Microbiota/*genetics ; Phylogeny ; Proteobacteria/genetics/isolation & purification ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Scotland ; Sex Factors ; Sympatry ; }, abstract = {The increasingly recognised effects of microbiomes on the eco-evolutionary dynamics of their hosts are promoting a view of the "hologenome" as an integral host-symbiont evolutionary entity. For example, sex-ratio distorting reproductive parasites such as Wolbachia are well-studied pivotal drivers of invertebrate reproductive processes, and more recent work is highlighting novel effects of microbiome assemblages on host mating behaviour and developmental incompatibilities that underpin or reinforce reproductive isolation processes. However, examining the hologenome and its eco-evolutionary effects in natural populations is challenging because microbiome composition is considerably influenced by environmental factors. Here we illustrate these challenges in a sympatric species complex of intertidal isopods (Jaera albifrons spp.) with pervasive sex-ratio distortion and ecological and behavioural reproductive isolation mechanisms. We deep-sequence the bacterial 16S rRNA gene among males and females collected in spring and summer from two coasts in north-east Scotland, and examine microbiome composition with a particular focus on reproductive parasites. Microbiomes of all species were diverse (overall 3,317 unique sequences among 3.8 million reads) and comprised mainly Proteobacteria and Bacteroidetes taxa typical of the marine intertidal zone, in particular Vibrio spp. However, we found little evidence of the reproductive parasites Wolbachia, Rickettsia, Spiroplasma and Cardinium, suggesting alternative causes of sex-ratio distortion. Notwithstanding, a significant proportion of the variance in microbiome composition among samples was explained by sex (14.1 %), nested within geographic (26.9 %) and seasonal (39.6 %) variance components. The functional relevance of this sex signal was difficult to ascertain given the absence of reproductive parasites, the ephemeral nature of the species assemblages and substantial environmental variability. These results establish the Jaera albifrons species complex as an intriguing system for examining the effects of microbiomes on reproductive processes and speciation, and highlight the difficulties associated with snapshot assays of microbiome composition in dynamic and complex environments.}, } @article {pmid30150735, year = {2019}, author = {Shaw, WR and Catteruccia, F}, title = {Vector biology meets disease control: using basic research to fight vector-borne diseases.}, journal = {Nature microbiology}, volume = {4}, number = {1}, pages = {20-34}, pmid = {30150735}, issn = {2058-5276}, support = {R01 AI124165/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/virology ; Animals ; Anopheles/microbiology/parasitology ; Biological Control Agents/pharmacology ; Communicable Disease Control/*methods ; Communicable Diseases/*transmission ; Culex/parasitology/virology ; Disease Outbreaks/*prevention & control ; Humans ; Insect Control/*methods ; Insecticide Resistance ; Insecticides/pharmacology ; Mosquito Vectors/*microbiology/*parasitology ; Wolbachia/pathogenicity ; }, abstract = {Human pathogens that are transmitted by insects are a global problem, particularly those vectored by mosquitoes; for example, malaria parasites transmitted by Anopheles species, and viruses such as dengue, Zika and chikungunya that are carried by Aedes mosquitoes. Over the past 15 years, the prevalence of malaria has been substantially reduced and virus outbreaks have been contained by controlling mosquito vectors using insecticide-based approaches. However, disease control is now threatened by alarming rates of insecticide resistance in insect populations, prompting the need to develop a new generation of specific strategies that can reduce vector-mediated transmission. Here, we review how increased knowledge in insect biology and insect-pathogen interactions is stimulating new concepts and tools for vector control. We focus on strategies that either interfere with the development of pathogens within their vectors or directly impact insect survival, including enhancement of vector-mediated immune control, manipulation of the insect microbiome, or use of powerful new genetic tools such as CRISPR-Cas systems to edit vector genomes. Finally, we offer a perspective on the implementation hurdles as well as the knowledge gaps that must be filled in the coming years to safely realize the potential of these novel strategies to eliminate the scourge of vector-borne disease.}, } @article {pmid30150426, year = {2018}, author = {Kostaropoulos, T and Papageorgiou, L and Champeris Tsaniras, S and Vlachakis, D and Eliopoulos, E}, title = {Carcinogenic Pesticide Control via Hijacking Endosymbiosis; The Paradigm of DSB-A from Wolbachia pipientis for the Management of Otiorhynchus singularis.}, journal = {In vivo (Athens, Greece)}, volume = {32}, number = {5}, pages = {1051-1062}, pmid = {30150426}, issn = {1791-7549}, mesh = {Amino Acid Motifs ; Amino Acid Sequence ; Animals ; *Carcinogens/toxicity ; *Chemical Safety ; Coleoptera/*microbiology ; Conserved Sequence ; Drug Design ; *Insect Control ; Models, Molecular ; *Pesticides/adverse effects ; Phylogeny ; Protein Conformation ; Protein Disulfide-Isomerases/chemistry/classification/genetics/*metabolism ; Structure-Activity Relationship ; *Symbiosis ; Wolbachia/enzymology/genetics/*metabolism ; }, abstract = {BACKGROUND/AIM: Pesticides have little, if any specificity, to the pathogen they target in most cases. Wide spectrum toxic chemicals are being used to remove pestcides and salvage crops and economies linked to agriculture. The burden on the environment, public health and economy is huge. Traditional pestcide control is based on administering heavy loads of highly toxic compounds and elements that essentially strip all life from the field. Those chemicals are a leading cause of increased cancer related deaths in countryside. Herein, the Trojan horse of endosymbiosis was used, in an effort to control pests using high specificity compounds in reduced quantities.

MATERIALS AND METHODS: Our pipeline has been applied on the case of Otiorhynchus singularis, which is a very widespread pest, whose impact is devastating on a repertoire of crops. To date, there is no specific pesticide nor agent to control it. The deployed strategy involves the inhibition of the key DSB-A enzyme of its endosymbiotic Wolbachia pipientis bacterial strain.

RESULTS: Our methodology, provides the means to design, test and identify highly specific pestcide control substances that minimize the impact of toxic chemicals on health, economy and the environment.

CONCLUSION: All in all, in this study a radical computer-based pipeline is proposed that could be adopted under many other similar scenarios and pave the way for precision agriculture via optimized pest control.}, } @article {pmid30149795, year = {2018}, author = {Brown, AMV}, title = {Endosymbionts of Plant-Parasitic Nematodes.}, journal = {Annual review of phytopathology}, volume = {56}, number = {}, pages = {225-242}, doi = {10.1146/annurev-phyto-080417-045824}, pmid = {30149795}, issn = {1545-2107}, mesh = {Animals ; *Bacterial Physiological Phenomena ; Host-Parasite Interactions ; Nematoda/*microbiology ; Plant Diseases/parasitology/*prevention & control ; Plants/*parasitology ; *Symbiosis ; }, abstract = {Some of the most agriculturally important plant-parasitic nematodes (PPNs) harbor endosymbionts. Extensive work in other systems has shown that endosymbionts can have major effects on host virulence and biology. This review highlights the discovery, development, and diversity of PPN endosymbionts, incorporating inferences from genomic data. Cardinium, reported from five PPN hosts to date, is characterized by its presence in the esophageal glands and other tissues, with a discontinuous distribution across populations, and genomic data suggestive of horizontal gene exchange. Xiphinematobacter occurs in at least 27 species of dagger nematode in the ovaries and gut epithelial cells, where genomic data suggest it may serve in nutritional supplementation. Wolbachia, reported in just three PPNs, appears to have an ancient history in the Pratylenchidae and displays broad tissue distribution and genomic features intermediate between parasitic and reproductive groups. Finally, a model is described that integrates these insights to explain patterns of endosymbiont replacement.}, } @article {pmid30148833, year = {2018}, author = {Cerutti, F and Modesto, P and Rizzo, F and Cravero, A and Jurman, I and Costa, S and Giammarino, M and Mandola, ML and Goria, M and Radovic, S and Cattonaro, F and Acutis, PL and Peletto, S}, title = {The microbiota of hematophagous ectoparasites collected from migratory birds.}, journal = {PloS one}, volume = {13}, number = {8}, pages = {e0202270}, pmid = {30148833}, issn = {1932-6203}, mesh = {Animal Migration ; Animals ; Arthropods/*microbiology ; Bacteria/*isolation & purification ; Bird Diseases/*parasitology ; Birds/parasitology ; Computational Biology ; Ectoparasitic Infestations/parasitology/*veterinary ; Italy ; *Microbiota ; Molecular Typing ; Parasites/*microbiology ; RNA, Ribosomal, 16S ; Ticks/microbiology ; }, abstract = {Arthropod vectors are responsible for the transmission of human pathogens worldwide. Several arthropod species are bird ectoparasites, however, no study to date has characterized their microbiota as a whole. We sampled hematophagous ectoparasites that feed on migratory birds and performed 16S rRNA gene metabarcoding to characterize their microbial community. A total of 194 ectoparasites were collected from 115 avian hosts and classified into three groups: a) Hippoboscidae diptera; b) ticks; c) other arthropods. Metabarcoding showed that endosymbionts were the most abundant genera of the microbial community, including Wolbachia for Hippoboscidae diptera, Candidatus Midichloria for ticks, Wolbachia and Arsenophonus for the other arthropod group. Genera including pathogenic species were: Rickettsia, Borrelia, Coxiella, Francisella, Bartonella, Anaplasma. Co-infection with Borrelia-Rickettsia and Anaplasma-Rickettsia was also observed. A global overview of the microbiota of ectoparasites sampled from migratory birds was obtained with the use of 16S rRNA gene metabarcoding. A novel finding is the first identification of Rickettsia in the common swift louse fly, Crataerina pallida. Given their possible interaction with pathogenic viruses and bacteria, the presence of endosymbionts in arthropods merits attention. Finally, molecular characterization of genera, including both pathogenic and symbiont species, plays a pivotal role in the design of targeted molecular diagnostics.}, } @article {pmid30139962, year = {2019}, author = {Richardson, KM and Griffin, PC and Lee, SF and Ross, PA and Endersby-Harshman, NM and Schiffer, M and Hoffmann, AA}, title = {A Wolbachia infection from Drosophila that causes cytoplasmic incompatibility despite low prevalence and densities in males.}, journal = {Heredity}, volume = {122}, number = {4}, pages = {428-440}, pmid = {30139962}, issn = {1365-2540}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Cytoplasm/*microbiology ; Drosophila/*genetics/*microbiology ; Female ; Fertility/genetics ; Male ; Phylogeny ; Reproduction ; Symbiosis/genetics ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Wolbachia bacteria are common insect endosymbionts transmitted maternally and capable of spreading through insect populations by cytoplasmic incompatibility (CI) when infected males cause embryo death after mating with uninfected females. Selection in the Wolbachia endosymbiont occurs on female hosts and is expected to favour strong maternal transmission to female offspring, even at the cost of reduced CI. With maternal leakage, nuclear genes are expected to be selected to suppress cytoplasmic incompatibility caused by males while also reducing any deleterious effects associated with the infection. Here we describe a new type of Wolbachia strain from Drosophila pseudotakahashii likely to have arisen from evolutionary processes on host and/or Wolbachia genomes. This strain is often absent from adult male offspring, but always transmitted to females. It leads to males with low or non-detectable Wolbachia that nevertheless show CI. When detected in adult males, the infection has a low density relative to that in females, a phenomenon not previously seen in Wolbachia infections of Drosophila. This Wolbachia strain is common in natural populations, and shows reduced CI when older (infected) males are crossed. These patterns highlight that endosymbionts can have strong sex-specific effects and that high frequency Wolbachia strains persist through effects on female reproduction. Female-limited Wolbachia infections may be of applied interest if the low level of Wolbachia in males reduces deleterious fitness effects on the host.}, } @article {pmid30124918, year = {2018}, author = {Ali, H and Muhammad, A and Sanda Bala, N and Hou, Y}, title = {The Endosymbiotic Wolbachia and Host COI Gene Enables to Distinguish Between Two Invasive Palm Pests; Coconut Leaf Beetle, Brontispa longissima and Hispid Leaf Beetle, Octodonta nipae.}, journal = {Journal of economic entomology}, volume = {111}, number = {6}, pages = {2894-2902}, pmid = {30124918}, issn = {1938-291X}, mesh = {Animals ; Base Sequence ; Coleoptera/*classification/genetics/microbiology ; Electron Transport Complex IV/genetics ; Species Specificity ; Wolbachia/genetics ; }, abstract = {To elucidate taxonomic eminence of identical pest species is essential for many ecological and conservation studies. Without proficient skills, accurate molecular identification and characterization are laborious and time-consuming. The coconut leaf beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), is biologically and morphologically identical to hispid leaf beetle, Octodonta nipae (Maulik) (Coleoptera: Chrysomelidae), and is known as the most harming nuisances of palm cultivation worldwide. The present examination was to establish Wolbachia genotyping analysis along with host cytochrome oxidase subunit I (COI) gene for accurate identification between these individuals of the same family (Chrysomelidae). Here, we have cloned and sequenced a gene coding Wolbachia surface protein (wsp) and COI gene regions amplified from both species by polymerase chain reaction. The nucleotide sequences were directly determined (≈600 bp for wsp and ≈804 bp for COI) and aligned using the multiple alignment algorithms in the ESPript3 package and the MEGA5 program. Comparative sequence analysis indicated that the representative of wsp and COI sequences from these two beetles were highly variable. To ensure this bacterial variation, multilocus sequence typing (MLST) of bacterial genes was conducted, and the results vindicated the same trend of variations. Furthermore, the phylogenetic analysis also indicates that B. longissima and O. nipae being the two different species harbors two distinct Wolbachia Hertig and Burt (Rickettsiales: Anaplamataceae) supergroups B and A, respectively. The present outcomes quickly discriminate between these two species. Considering its simplicity and cost-effectiveness, it can be used as a diagnostic tool for discriminating such invasive species particularly B. longissima and O. nipae which has overlapping morphologic characters.}, } @article {pmid30113326, year = {2018}, author = {Showler, AJ and Nutman, TB}, title = {Imported onchocerciasis in migrants and travelers.}, journal = {Current opinion in infectious diseases}, volume = {31}, number = {5}, pages = {393-398}, doi = {10.1097/QCO.0000000000000483}, pmid = {30113326}, issn = {1473-6527}, mesh = {Animals ; Antibodies, Helminth/blood ; Communicable Diseases, Imported/*diagnosis/*drug therapy ; Diagnostic Tests, Routine/*methods ; *Disease Management ; Doxycycline/therapeutic use ; Filaricides/*therapeutic use ; Human Migration ; Humans ; Onchocerca volvulus/immunology ; Onchocerciasis/*diagnosis/*drug therapy ; Transients and Migrants ; Travel ; }, abstract = {PURPOSE OF REVIEW: With increasing international travel and mass global population migration, clinicians in nonendemic countries must be familiar with imported neglected tropical diseases including onchocerciasis, which is commonly known as 'river blindness'.

RECENT FINDINGS: Imported onchocerciasis manifests differently in travelers compared with migrants from endemic areas and is likely underdiagnosed in both groups. Recent clinical studies confirm that eosinophilia is not a sensitive marker for Onchocerca volvulus, with one-third of patients having a normal eosinophil count. Novel diagnostics measuring antibodies to multiple recombinant O. volvulus antigens maintain a high sensitivity while improving specificity compared with conventional pan-filarial serologic testing. A 6-week course of doxycycline has macrofilaricidal activity through Wolbachia depletion and may be useful in nonendemic areas in addition to standard serial ivermectin.

SUMMARY: Recent studies characterizing distinct clinical presentations in travelers and migrants may enable clinicians to better recognize imported onchocerciasis. Although novel diagnostics have improved specificity, most remain restricted to tropical disease reference laboratories and to date there is no marker of cure. Prolonged doxycycline treatment may reduce the need for serial ivermectin, though more potent short-course macrofilaricidal drugs are being developed.}, } @article {pmid30110391, year = {2018}, author = {Russell, SL and Lemseffer, N and White, PM and Sullivan, WT}, title = {Wolbachia and host germline components compete for kinesin-mediated transport to the posterior pole of the Drosophila oocyte.}, journal = {PLoS pathogens}, volume = {14}, number = {8}, pages = {e1007216}, pmid = {30110391}, issn = {1553-7374}, mesh = {Animals ; *Binding, Competitive ; Cell Polarity ; Drosophila melanogaster/metabolism/*microbiology ; Germ Cells/microbiology ; *Host-Pathogen Interactions/physiology ; Kinesins/*metabolism ; Microtubules/metabolism ; Oocytes/metabolism/*microbiology ; Oogenesis/physiology ; Protein Transport ; Tissue Distribution ; Wolbachia/metabolism/*physiology ; }, abstract = {Widespread success of the intracellular bacterium Wolbachia across insects and nematodes is due to efficient vertical transmission and reproductive manipulations. Many strains, including wMel from Drosophila melanogaster, exhibit a specific concentration to the germplasm at the posterior pole of the mature oocyte, thereby ensuring high fidelity of parent-offspring transmission. Transport of Wolbachia to the pole relies on microtubules and the plus-end directed motor kinesin heavy chain (KHC). However, the mechanisms mediating Wolbachia's association with KHC remain unknown. Here we show that reduced levels of the host canonical linker protein KLC results in dramatically increased levels of Wolbachia at the oocyte's posterior, suggesting that KLC and some key associated host cargos outcompete Wolbachia for association with a limited amount of KHC motor proteins. Consistent with this interpretation, over-expression of KHC causes similarly increased levels of posteriorly localized Wolbachia. However, excess KHC has no effect on levels of Vasa, a germplasm component that also requires KHC for posterior localization. Thus, Wolbachia transport is uniquely KHC-limited because these bacteria are likely outcompeted for binding to KHC by some host cargo/linker complexes. These results reveal a novel host-symbiont interaction that underscores the precise regulation required for an intracellular bacterium to co-opt, but not disrupt, vital host processes.}, } @article {pmid30107579, year = {2018}, author = {Flatau, R and Segoli, M and Khokhlova, I and Hawlena, H}, title = {Wolbachia's role in mediating its flea's reproductive success differs according to flea origin.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {10}, pages = {}, doi = {10.1093/femsec/fiy157}, pmid = {30107579}, issn = {1574-6941}, mesh = {Animals ; Host Specificity ; Models, Biological ; Reproduction ; Rodentia/parasitology ; Selection, Genetic ; Siphonaptera/growth & development/*microbiology/*physiology ; Symbiosis ; Wolbachia/growth & development/*physiology ; }, abstract = {Endosymbionts-microbes that live within and engage in prolonged and intimate associations with their hosts-are gaining recognition for their direct impact on plant and animal reproduction. Here we used the overlooked Wolbachia-flea system to explore the possibility that endosymbionts may also play a role as mediators in shaping the reproductive success of their hosts. We simultaneously quantified the Wolbachia density in field- and laboratory-originated fleas that fed and mated on rodents for either 5 or 10 days and assessed their body size and current reproductive success. By combining multigroup analysis and model selection approaches, we teased apart the contribution of the direct effects of the flea's physiological age and body size and the mediation effect of its Wolbachia endosymbionts on flea reproductive success, and we showed that the latter was stronger than the former. However, interestingly, the mediation effect was manifested only in laboratory-originated fleas, for which the increase in Wolbachia with age translated into lower reproductive success. These results suggest that some well-supported phenomena, such as aging effects, may be driven by endosymbionts and show once again that the role of endosymbionts in shaping the reproductive success of their host depends on their selective environment.}, } @article {pmid30106293, year = {2018}, author = {Kruse, A and Ramsey, JS and Johnson, R and Hall, DG and MacCoss, MJ and Heck, M}, title = {Candidatus Liberibacter asiaticus Minimally Alters Expression of Immunity and Metabolism Proteins in Hemolymph of Diaphorina citri, the Insect Vector of Huanglongbing.}, journal = {Journal of proteome research}, volume = {17}, number = {9}, pages = {2995-3011}, doi = {10.1021/acs.jproteome.8b00183}, pmid = {30106293}, issn = {1535-3907}, mesh = {Acetylation ; Animals ; Bacterial Proteins/classification/genetics/*metabolism ; Citrus/parasitology ; Energy Metabolism ; Fatty Acids ; Gene Ontology ; Hemiptera/genetics/immunology/*metabolism/microbiology ; Hemolymph/*chemistry/immunology/metabolism/microbiology ; Host-Pathogen Interactions/genetics/immunology ; Insect Proteins/classification/genetics/immunology/*metabolism ; Insect Vectors/genetics/immunology/metabolism/microbiology ; Lipid Metabolism ; Molecular Chaperones/genetics/metabolism ; Molecular Sequence Annotation ; Phosphorylation ; Plant Diseases/parasitology ; *Protein Processing, Post-Translational ; Proteome/classification/genetics/immunology/*metabolism ; Proteomics/methods ; Rhizobiaceae/genetics/*metabolism ; Symbiosis/genetics/immunology ; Vitellogenins ; Wolbachia/genetics/metabolism ; }, abstract = {Huanglongbing (HLB), also known as citrus greening disease, is the most serious disease of citrus plants. It is associated with the Gram-negative bacterium ' Candidatus Liberibacter asiaticus' (CLas), which is transmitted between host plants by the hemipteran insect vector Diaphorina citri in a circulative, propagative manner involving specific interactions with various insect tissues including the hemolymph, fluid that occupies the body cavity akin to insect blood. High resolution quantitative mass spectrometry was performed to investigate the effect of CLas exposure on D. citri hemolymph at the proteome level. In contrast to the broad proteome effects on hundreds of proteins and a diverse array of metabolic pathways previously reported in gut and whole insect proteome analyses, the effect of CLas on the hemolymph was observed to be highly specific, restricted to key immunity and metabolism pathways, and lower in magnitude than that previously observed in the whole insect body and gut. Vitellogenins were abundantly expressed and CLas-responsive. Gene-specific RNA expression analysis suggests that these proteins are expressed in both male and female insects and may have roles outside of reproductive vitellogenesis. Proteins for fatty acid synthesis were found to be up-regulated, along with metabolic proteins associated with energy production, supported at the organismal level by the previously published observation that D. citri individuals experience a higher level of hunger when reared on CLas-infected plants. Prediction of post-translational modifications identified hemolymph proteins with phosphorylation and acetylation upon CLas exposure. Proteins derived from the three most prominent bacterial endosymbionts of the psyllid were also detected in the hemolymph, and several of these have predicted secretion signals. A DNAK protein, the bacterial HSP70, detected in the hemolymph expressed from Wolbachia pipientis was predicted to encode a eukaryotic nuclear localization signal. Taken together, these data show specific changes to immunity and metabolism in D. citri hemolymph involving host and endosymbiont proteins. These data provide a novel context for proteomic changes seen in other D. citri tissues in response to CLas and align with organismal data on the effects of CLas on D. citri metabolism and reproduction.}, } @article {pmid30104608, year = {2018}, author = {Mukherjee, S and Joardar, N and Mondal, S and Schiefer, A and Hoerauf, A and Pfarr, K and Babu, SPS}, title = {Quinolone-fused cyclic sulfonamide as a novel benign antifilarial agent.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {12073}, pmid = {30104608}, issn = {2045-2322}, mesh = {Aedes ; Animals ; Apoptosis/drug effects ; Cattle ; Disease Models, Animal ; Elephantiasis, Filarial/*drug therapy/parasitology ; Female ; Filaricides/chemistry/*pharmacology/therapeutic use ; Humans ; Male ; Mice ; Oxidative Stress/drug effects ; Parasitic Sensitivity Tests ; Quinolones/chemistry/*pharmacology/therapeutic use ; RAW 264.7 Cells ; Rats ; Reactive Oxygen Species/metabolism ; Setaria Nematode/*drug effects/metabolism ; Sulfonamides/chemistry/*pharmacology/therapeutic use ; Wolbachia/drug effects/metabolism ; }, abstract = {Search of potent antifilarial drugs has been a major thrust area in tropical medicine research over the decades. Herein, we report 4,7-dimethyl-3,4,7,8-tetrahydro-3λ[6]-[1,2]thiazino[4,3-f]quinoline-3,3,8-trione (8l) as a new class of antifilarial agent which is extremely potent, with lethality against all the developmental stages (oocyte, microfilaria and adult) of the filarial parasite Setaria cervi. Molecular investigation on its mode of action revealed that 8l is a typical inducer of reactive oxygen species that triggers oxidative stress inside the filarid and further signals induction of apoptosis by activating both intrinsic and extrinsic pathways. Moreover, 8l is also active against Wolbachia, the essential endosymbiont of several human infectious filarids. Selective toxicity against filarial parasites and non-toxic nature in rat model were found as unique traits of 8l to be a future medicine. Taken en masse, this maiden report on a novel quinolone fused cyclic sulfonamide presents a promising therapeutic lead for lymphatic filariasis in future.}, } @article {pmid30092356, year = {2018}, author = {Jiang, W and Zhu, J and Wu, Y and Li, L and Li, Y and Ge, C and Wang, Y and Endersby, NM and Hoffmann, AA and Yu, W}, title = {Influence of Wolbachia infection on mitochondrial DNA variation in the genus Polytremis (Lepidoptera: Hesperiidae).}, journal = {Molecular phylogenetics and evolution}, volume = {129}, number = {}, pages = {158-170}, doi = {10.1016/j.ympev.2018.08.001}, pmid = {30092356}, issn = {1095-9513}, mesh = {Animals ; Cell Nucleus/genetics ; China ; DNA, Mitochondrial/*genetics ; Female ; *Genetic Variation ; Geography ; Haplotypes/genetics ; Lepidoptera/*genetics/*microbiology ; Likelihood Functions ; Male ; Multilocus Sequence Typing ; Phylogeny ; Population Density ; Time Factors ; Wolbachia/*physiology ; }, abstract = {The maternally inherited obligate bacteria Wolbachia is known for infecting the reproductive tissues of a wide range of arthropods and can contribute to phylogenetically discordant patterns between mtDNA and nDNA. In this study, we tested for an association between mito-nuclear discordance in Polytremis and Wolbachia infection. Six of the 17 species of Polytremis were found to be infected with Wolbachia. Overall, 34% (70/204) of Polytremis specimens were Wolbachia positive and three strains of Wolbachia identified using a wsp marker were further characterized as six strains based on MLST markers. Wolbachia acquisition in Polytremis appears to occur mainly through horizontal transmission rather than codivergence based on comparison of the divergence times of Wolbachia and Polytremis species. At the intraspecific level, one of the Wolbachia infections (wNas1) is associated with reduced mtDNA polymorphism in the infected Polytremis population. At the interspecific level, there is one case of mito-nuclear discordance likely caused by introgression of P. fukia mtDNA into P. nascens driven by another Wolbachia strain (wNas3). Based on an absence of infected males, we suspect that one Wolbachia strain (wNas2) affects sex ratio, but the phenotypic effects of the other strains are unclear. These data reveal a dynamic interaction between Polytremis and Wolbachia endosymbionts affecting patterns of mtDNA variation.}, } @article {pmid30071091, year = {2018}, author = {Johnson, BJ and Rohde, BB and Zeak, N and Staunton, KM and Prachar, T and Ritchie, SA}, title = {A low-cost, battery-powered acoustic trap for surveilling male Aedes aegypti during rear-and-release operations.}, journal = {PloS one}, volume = {13}, number = {8}, pages = {e0201709}, pmid = {30071091}, issn = {1932-6203}, mesh = {*Acoustics ; *Aedes ; Animals ; *Costs and Cost Analysis ; *Electric Power Supplies ; Male ; Mosquito Control/*economics/*instrumentation ; }, abstract = {The Aedes aegypti mosquito is a primary vector of several serious arboviruses throughout the world and is therefore of great concern to many public health organizations. With vector control methodology pivoting towards rearing and releasing large numbers of genetically modified, sterilized, or Wolbachia-infected male mosquitoes to control vector populations, economical surveillance methods for release tracking becomes increasingly necessary. Previous work has identified that male Ae. aegypti are attracted to female wingbeat frequencies and can be captured through artificial playback of these frequencies, but the tested systems are cost-prohibitive for wide-scale monitoring. Thus, we have developed a simple, low-cost, battery-powered, microcontroller-based sound lure which mimics the wingbeat frequency of female Ae. aegypti, thereby attracting males. We then tested the efficacy of this lure in combination with a passive (non-powered) gravid Aedes trap (GAT) against the current gold-standard, the Biogents Sentinel (BGS) trap, which requires main power (household power) and costs several times what the GAT does. Capture rates of male Ae. aegypti in sound-baited GATs (Sound-GATs) in these field tests were comparable to that of the BGS with no inhibitory effects of sound playback on female capture. We conclude that the Sound-GAT is an effective replacement of the costly BGS for surveillance of male Ae. aegypti mosquitoes, particularly in the developing countries where funding is limited, and has the potential to be adapted to target males of other medically important species.}, } @article {pmid30069620, year = {2018}, author = {Fallon, AM}, title = {Strain-specific response to ampicillin in Wolbachia-infected mosquito cell lines.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {54}, number = {8}, pages = {580-588}, pmid = {30069620}, issn = {1543-706X}, support = {AI081322//National Institutes of Health/ ; }, mesh = {Aedes/drug effects/genetics/growth & development/*microbiology ; Amino Acid Sequence ; Ampicillin/*pharmacology ; Animals ; Cell Line ; Gene Expression Regulation/drug effects ; Penicillin-Binding Proteins/genetics/metabolism ; Peptides/chemistry ; Sequence Alignment ; Species Specificity ; Tandem Mass Spectrometry ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis (Rickettsiales; Anaplasmataceae) is an obligate intracellular alpha proteobacterium that occurs in arthropods and filarial worms. Some strains of Wolbachia can be maintained as persistent infections in insect cell lines. C/wStr1 cells from the mosquito Aedes albopictus maintain a robust infection with Wolbachia strain wStr, originally isolated from the planthopper, Laodelphax striatellus. To explore possible functions of penicillin-binding proteins expressed from the wStr genome, C/wStr1 cells were exposed to ampicillin. Absolute levels of Wolbachia increased 3.5-fold in ampicillin-treated cells and fivefold in naive cells newly infected with wStr. Because cell numbers were depressed by ampicillin treatment, Wolbachia yield on a per-cell basis increased by 15-fold. The absence of a similar effect on wAlbB in Aa23 host cells suggests that the Wolbachia strain, the presence/absence of genes encoding penicillin-binding proteins, or the interaction between wAlbB and its host cells may modulate the effects of ampicillin.}, } @article {pmid30068363, year = {2018}, author = {Callahan, AG and Ross, PA and Hoffmann, AA}, title = {Small females prefer small males: size assortative mating in Aedes aegypti mosquitoes.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {445}, pmid = {30068363}, issn = {1756-3305}, support = {1037003//National Health and Medical Research Council/ ; }, mesh = {Aedes/*anatomy & histology/physiology ; Animals ; *Body Size ; Female ; Male ; Sexual Behavior, Animal/*physiology ; }, abstract = {BACKGROUND: With Aedes aegypti mosquitoes now being released in field programmes aimed at disease suppression, there is interest in identifying factors influencing the mating and invasion success of released mosquitoes. One factor that can increase release success is size: released males may benefit competitively from being larger than their field counterparts. However, there could be a risk in releasing only large males if small field females avoid these males and instead prefer small males. Here we investigate this risk by evaluating mating success for mosquitoes differing in size.

RESULTS: We measured mating success indirectly by coupling size with Wolbachia-infected or uninfected mosquitoes and scoring cytoplasmic incompatibility. Large females showed no evidence of a mating preference, whereas small males were relatively more successful than large males when mating with small females, exhibiting an advantage of around 20-25%.

CONCLUSIONS: Because field females typically encompass a wide range of sizes while laboratory reared (and released) males typically fall into a narrow size range of large mosquitoes, these patterns can influence the success of release programmes which rely on cytoplasmic incompatibility to suppress populations and initiate replacement invasions. Releases could include some small males generated under low food or crowded conditions to counter this issue, although this would need to be weighed against issues associated with costs of producing males of various size classes.}, } @article {pmid30064911, year = {2018}, author = {Fukui, T and Kiuchi, T and Shoji, K and Kawamoto, M and Shimada, T and Katsuma, S}, title = {In vivo masculinizing function of the Ostrinia furnacalis Masculinizer gene.}, journal = {Biochemical and biophysical research communications}, volume = {503}, number = {3}, pages = {1768-1772}, doi = {10.1016/j.bbrc.2018.07.111}, pmid = {30064911}, issn = {1090-2104}, mesh = {Animals ; Female ; Insect Proteins/*genetics/metabolism ; Male ; Moths/embryology/*genetics ; Sex Chromosomes/*genetics/metabolism ; Sex Determination Processes/*genetics ; }, abstract = {The Masculinizer gene (Masc) encodes a CCCH tandem zinc finger protein essential for masculinization and dosage compensation in the silkworm Bombyx mori. Previously we identified a Masc orthologue from the crambid Ostrinia furnacalis (OfMasc) and observed its masculinizing activity in the B. mori cultured cell line BmN-4. However, the role of OfMasc in masculinization of O. furnacalis has not been assessed. In this study, we unexpectedly discovered that all of the male larvae that escaped from Wolbachia-induced embryonic male-killing by OfMasc cRNA injection expressed the female-type splicing variants of O. furnacalis doublesex (Ofdsx). To clarify the role of OfMasc in the masculinization process in vivo, we established a system to monitor both sex chromosome- and dsx splicing-based sexes from a single O. furnacalis embryo. Using this system, we investigated the effects of OfMasc knockdown in early embryos on Ofdsx splicing and found that depletion of OfMasc mRNA in male embryos induced the production of the female-type splicing variants of Ofdsx. This result indicates that OfMasc is required for masculinization in O. furnacalis, and that the Masc protein possesses masculinizing activity in an insect species that is phylogenetically distant from Bombycidae.}, } @article {pmid30061694, year = {2018}, author = {Fisher, ML and Watson, DW and Osborne, JA and Mochizuki, H and Breen, M and Schal, C}, title = {Growth kinetics of endosymbiont Wolbachia in the common bed bug, Cimex lectularius.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {11444}, pmid = {30061694}, issn = {2045-2322}, support = {P30 ES025128/ES/NIEHS NIH HHS/United States ; NCHHU0017-13//U.S. Department of Housing and Urban Development (HUD)/International ; 2013-5-35 MBE//Alfred P. Sloan Foundation/International ; }, mesh = {Animals ; Bedbugs/*microbiology ; DNA/genetics ; Female ; Kinetics ; Larva/microbiology ; *Symbiosis ; Wolbachia/*growth & development ; }, abstract = {The common bed bug, Cimex lectularius harbors the endosymbiotic microorganism, Wolbachia (wCle), in a gonad-associated bacteriome as an obligate nutritional mutualist. The obligatory nature of this association suggests that all individuals in C. lectularius populations would be infected with wCle. However, studies spanning the past several decades have reported variation in both infection frequency and relative abundance of wCle in field-collected samples of bed bugs. Since the growth kinetics of wCle is poorly understood, the objective of this study was to quantify wCle over the life cycle of two strains of C. lectularius. Our results highlight that wCle is dynamic during bed bug development, changing relative to life stage, intermolt stage, and blood-fed status. These results suggest new hypotheses about the coordination of Wolbachia growth and regression with its host's physiology and endocrine events. The observed quantitative modulation of wCle during the bed bug life cycle and during periods of starvation may explain the disparities in wCle infections reported in field-collected C. lectularius.}, } @article {pmid30060072, year = {2018}, author = {Gillespie, JJ and Driscoll, TP and Verhoeve, VI and Rahman, MS and Macaluso, KR and Azad, AF}, title = {A Tangled Web: Origins of Reproductive Parasitism.}, journal = {Genome biology and evolution}, volume = {10}, number = {9}, pages = {2292-2309}, pmid = {30060072}, issn = {1759-6653}, support = {R01 AI017828/AI/NIAID NIH HHS/United States ; R01 AI122672/AI/NIAID NIH HHS/United States ; R01 AI126853/AI/NIAID NIH HHS/United States ; R21 AI126108/AI/NIAID NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Arthropods/*microbiology/physiology ; Bacterial Proteins/chemistry/genetics ; Bacterial Toxins/chemistry/genetics ; Biological Evolution ; Female ; Male ; Models, Molecular ; Phylogeny ; Plasmids/genetics ; Reproduction ; Rickettsia felis/chemistry/*genetics/physiology ; Wolbachia/chemistry/*genetics/physiology ; }, abstract = {While typically a flea parasite and opportunistic human pathogen, the presence of Rickettsia felis (strain LSU-Lb) in the non-blood-feeding, parthenogenetically reproducing booklouse, Liposcelis bostrychophila, provides a system to ascertain factors governing not only host transitions but also obligate reproductive parasitism (RP). Analysis of plasmid pLbAR, unique to R. felis str. LSU-Lb, revealed a toxin-antitoxin module with similar features to prophage-encoded toxin-antitoxin modules utilized by parasitic Wolbachia strains to induce another form of RP, cytoplasmic incompatibility, in their arthropod hosts. Curiously, multiple deubiquitinase and nuclease domains of the large (3,841 aa) pLbAR toxin, as well the entire antitoxin, facilitated the detection of an assortment of related proteins from diverse intracellular bacteria, including other reproductive parasites. Our description of these remarkable components of the intracellular mobilome, including their presence in certain arthropod genomes, lends insight on the evolution of RP, while invigorating research on parasite-mediated biocontrol of arthropod-borne viral and bacterial pathogens.}, } @article {pmid30059498, year = {2018}, author = {Xue, L and Fang, X and Hyman, JM}, title = {Comparing the effectiveness of different strains of Wolbachia for controlling chikungunya, dengue fever, and zika.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {7}, pages = {e0006666}, pmid = {30059498}, issn = {1935-2735}, support = {U01 GM097661/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology/virology ; Animals ; Chikungunya Fever/*transmission/virology ; Chikungunya virus/physiology ; Dengue/*transmission/virology ; Dengue Virus/physiology ; Female ; Humans ; Male ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology/physiology/virology ; Pest Control, Biological/*methods ; Reproduction ; Wolbachia/classification/*physiology ; Zika Virus/physiology ; Zika Virus Infection/*transmission/virology ; }, abstract = {Once Aedes aegypti and Aedes albopictus mosquitoes that spread Chikungunya virus, dengue virus, and Zika virus are infected with Wolbachia, they have reduced egg laying rates, reduced transmission abilities, and shorter lifespans. Since most infected mosquitoes are only infectious in the last few days of their lives, shortening a mosquito's lifespan by a day or two can greatly reduce their abilities to spread mosquito-borne viral diseases, such as Chikungunya, dengue fever, and Zika. We developed a mathematical model to compare the effectiveness of the wMel and wAlbB strains of Wolbachia for controlling the spread of these viruses. The differences among the diseases, mosquitoes, and Wolbachia strains are captured by the model parameters for the mosquito-human transmission cycle. Moreover, the model accounts for the behavior changes of infectious population created by differences in the malaise caused by these viruses. We derived the effective and basic reproduction numbers for the model that are used to estimate the number of secondary infections from the infectious populations. In the same density of Wolbachia-free Aedes aegypti or Aedes albopictus mosquitoes, we observed that wMel and wAlbB strains of Wolbachia can reduce the transmission rates of these diseases effectively.}, } @article {pmid30056075, year = {2018}, author = {Ciuca, L and Simòn, F and Rinaldi, L and Kramer, L and Genchi, M and Cringoli, G and Acatrinei, D and Miron, L and Morchon, R}, title = {Seroepidemiological survey of human exposure to Dirofilaria spp. in Romania and Moldova.}, journal = {Acta tropica}, volume = {187}, number = {}, pages = {169-174}, doi = {10.1016/j.actatropica.2018.07.012}, pmid = {30056075}, issn = {1873-6254}, mesh = {Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antibodies, Helminth/*immunology ; Antigens, Helminth/*immunology ; Child ; Dirofilaria/*immunology ; Dirofilaria immitis/immunology ; Dirofilaria repens/immunology ; Dirofilariasis/*epidemiology/immunology ; Dog Diseases/epidemiology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Moldova/epidemiology ; Prevalence ; Romania/epidemiology ; Seroepidemiologic Studies ; Surveys and Questionnaires ; Wolbachia/immunology ; Young Adult ; }, abstract = {The present study aimed to evaluate the extent of Dirofilaria immitis and D. repens exposure in humans from eastern and southern areas of Romania and central Moldova by serological methods. The serological screening was performed on a total of 450 serum samples (187 from Romania and 263 from Moldova). The sera were collected using a convenience sampling with the help of physicians from the hospitals of the study areas. All samples were analysed by a non-commercial ELISA test for the detection of IgG antibodies against adult somatic antigens of D. immitis and D. repens. The results showed a total of 49 (10.9%; 95% CI = 8.3-14.1%) individuals from Romania and Moldova with a positive response to IgG antibodies against both adult somatic antigens of D. immitis and D. repens. Specifically, 48 (10.7%; 95% CI = 8.0-14.0%) patients were positive for IgG-antibodies against adult somatic antigens of D. immitis, one (0.2%; 95% CI = 0.4-1.2%) against D. repens antigens, and four (0.9%; 95% CI = 0.4-3.3%). were positive for antigens of both parasites. At country level, out of 187 samples from Romania, 13 (6.9%; 95% CI = 4.1-11.5%) were positive for anti-D. immitis IgG with high exposure in the southern part of the country (Bucharest). Of the 263 people from Moldova, 36 (13.7%; 95% CI = 10.0-18.4%) were positive for D. immitis antigens from which three (1.1%, 95% CI = 0.4-3.3%) were positive for the antibodies against antigens of both parasites. Only one sample was found positive for anti-D. repens IgG. Positive IgG-ELISA results were confirmed by Western blot analysis. In addition, for further confirmation, a complementary ELISA was performed for anti-WSP IgG antibodies against Wolbachia endosymbionts. Our findings showed a noticeable exposure of humans from Romania and Moldova to Dirofilaria parasites. Serology can be useful for indicating exposure to Dirofilaria spp. in a healthy population in order to obtain useful data on the epidemiological scenario of human dirofilariosis in Eastern Europe.}, } @article {pmid30051873, year = {2018}, author = {Shi, M and White, VL and Schlub, T and Eden, JS and Hoffmann, AA and Holmes, EC}, title = {No detectable effect of Wolbachia wMel on the prevalence and abundance of the RNA virome of Drosophila melanogaster.}, journal = {Proceedings. Biological sciences}, volume = {285}, number = {1883}, pages = {}, pmid = {30051873}, issn = {1471-2954}, mesh = {Animals ; Australia ; Drosophila melanogaster/*microbiology/virology ; RNA Viruses/*physiology ; RNA, Viral/*analysis ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {Wolbachia is an endosymbiotic bacterium that can block viral infections in arthropods, generating interest in its potential to control the spread of mosquito-borne disease. Drosophila melanogaster is model organism for Wolbachia infection, and the wMel strain of Wolbachia can improve host survival following viral infection. However, it is unclear whether wMel induces anti-viral blocking against the broader native virome of D. melanogaster, or whether the major effect of Wolbachia is a reduction in viral abundance rather than viral clearance. We examined the effect of Wolbachia on viral abundance by comparing the total transcriptome of wMel-positive and wMel-negative D. melanogaster populations sampled from six locations in Australia. In addition, we examined the impact of wMel on individual flies by obtaining transcriptome data from 20 wMel-positive and 20 wMel-negative D. melanogaster from the location (Melbourne) with highest density of wMel. These data revealed high viral abundance in both Wolbachia-positive and -negative populations and individuals. Notably, none of the viral species identified, representing RNA viruses from at least nine families/floating genera, showed evidence of protection by wMel. Although the viral loads of picorna-like viruses are reduced by wMel under experimental conditions, we observed no such effect here. These data show that D. melanogaster can harbour abundant RNA viruses regardless of its Wolbachia status and imply that the interaction between Wolbachia and viruses in nature is more complex than simple blocking.}, } @article {pmid30050351, year = {2018}, author = {Ta-Tang, TH and Crainey, JL and Post, RJ and Luz, SL and Rubio, JM}, title = {Mansonellosis: current perspectives.}, journal = {Research and reports in tropical medicine}, volume = {9}, number = {}, pages = {9-24}, pmid = {30050351}, issn = {1179-7282}, abstract = {Mansonellosis is a filarial disease caused by three species of filarial (nematode) parasites (Mansonella perstans, Mansonella streptocerca, and Mansonella ozzardi) that use humans as their main definitive hosts. These parasites are transmitted from person to person by bloodsucking females from two families of flies (Diptera). Biting midges (Ceratopogonidae) transmit all three species of Mansonella, but blackflies (Simuliidae) are also known to play a role in the transmission of M. ozzardi in parts of Latin America. M. perstans and M. streptocerca are endemic in western, eastern, and central Africa, and M. perstans is also present in the neotropical region from equatorial Brazil to the Caribbean coast. M. ozzardi has a patchy distribution in Latin America and the Caribbean. Mansonellosis infections are thought to have little pathogenicity and to be almost always asymptomatic, but occasionally causing itching, joint pains, enlarged lymph glands, and vague abdominal symptoms. In Brazil, M. ozzardi infections are also associated with corneal lesions. Diagnosis is usually performed by detecting microfilariae in peripheral blood or skin without any periodicity. There is no standard treatment at present for mansonellosis. The combination therapy of diethylcarbamazine plus mebendazole for M. perstans microfilaremia is presently one of the most widely used, but the use of ivermectin has also been proven to be very effective against microfilariae. Recently, doxycycline has shown excellent efficacy and safety when used as an antimicrobial against endosymbiotic Wolbachia bacteria harbored by some strains of M. perstans and M. ozzardi. Diethylcarbamazine and ivermectin have been used effectively to treat M. streptocerca infection. There are at present no estimates of the disease burden caused by mansonellosis, and thus its importance to many global health professionals and policy makers is presently limited to how it can interfere with diagnostic tools used in modern filarial disease control and elimination programs aimed at other species of filariae.}, } @article {pmid30046071, year = {2018}, author = {Ferguson, NM}, title = {Challenges and opportunities in controlling mosquito-borne infections.}, journal = {Nature}, volume = {559}, number = {7715}, pages = {490-497}, pmid = {30046071}, issn = {1476-4687}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Dengue/mortality/*prevention & control/transmission ; Gene Drive Technology ; Goals ; Humans ; Incidence ; Malaria/mortality/*prevention & control/transmission ; Mosquito Control/*methods ; Mosquito Vectors/genetics/microbiology ; Pest Control, Biological/methods ; Vaccines ; Wolbachia/pathogenicity ; }, abstract = {Mosquito-borne diseases remain a major cause of morbidity and mortality across the tropical regions. Despite much progress in the control of malaria, malaria-associated morbidity remains high, whereas arboviruses-most notably dengue-are responsible for a rising burden of disease, even in middle-income countries that have almost completely eliminated malaria. Here I discuss how new interventions offer the promise of considerable future reductions in disease burden. However, I emphasize that intervention programmes need to be underpinned by rigorous trials and quantitative epidemiological analyses. Such analyses suggest that the long-term goal of elimination is more feasible for dengue than for malaria, even if malaria elimination would offer greater overall health benefit to the public.}, } @article {pmid30032285, year = {2018}, author = {Ahmadi, S and Poorjavad, N}, title = {Behavioral and Biological Effects of Exposure to Tuta absoluta (Lepidoptera: Gelechiidae) Sex Pheromone on Several Trichogramma (Hymenoptera: Trichogrammatidae) Populations.}, journal = {Journal of economic entomology}, volume = {111}, number = {6}, pages = {2667-2675}, doi = {10.1093/jee/toy212}, pmid = {30032285}, issn = {1938-291X}, mesh = {Animals ; Female ; Male ; Moths/chemistry/*parasitology ; Sex Attractants/*pharmacology ; Sexual Behavior, Animal/*drug effects ; Wasps/microbiology/*physiology ; Wolbachia/physiology ; }, abstract = {Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the most destructive pests of tomato worldwide. Biological control of the pest using Trichogramma (Hymenoptera: Trichogrammatidae) wasps can be combined with other practices such as use of synthetic sex pheromones for mating disruption or mass trapping programs. In this study, effects of T. absoluta sex pheromone on behavioral responses and fertility life table parameters of Trichogramma evanescens Westwood (Hymenoptera: Trichogrammatidae), Wolbachia-uninfected (W-) Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae), and Wolbachia-infected (W+) T. brassicae were investigated under laboratory conditions. Female wasps of T. evanescens and T. brassicae (W+) were attracted to the pest synthetic sex pheromone (around 62%) and calling virgin female moths (around 75%) and their responses were affected by the wasp age and temperature. Exposure to the host synthetic sex pheromone significantly reduced the percentage of adult emergence, longevity, and fecundity of female wasps, as well as the time spent to find a mate and duration of mating. However, tested populations were not affected similarly. Despite the laboratory tests, greenhouse experiments showed that the wasps were not caught in the traps baited with T. absoluta synthetic sex pheromone. Complementary studies are needed to precisely determine possible interference between pheromone application and releasing Trichogramma Westwood (Hymenoptera: Trichogrammatidae) wasps to achieve a successful integrated control of T. absoluta.}, } @article {pmid30025503, year = {2018}, author = {Zheng, B and Yu, J}, title = {Characterization of Wolbachia enhancing domain in mosquitoes with imperfect maternal transmission.}, journal = {Journal of biological dynamics}, volume = {12}, number = {1}, pages = {596-610}, doi = {10.1080/17513758.2018.1499969}, pmid = {30025503}, issn = {1751-3766}, mesh = {Animals ; Culicidae/*microbiology ; Dengue/*parasitology/*transmission ; Female ; *Infectious Disease Transmission, Vertical ; Models, Biological ; Pest Control, Biological ; Wolbachia/*physiology ; }, abstract = {A novel method to reduce the burden of dengue is to seed wild mosquitoes with Wolbachia-infected mosquitoes in dengue-endemic areas. Concerns in current mathematical models are to locate the Wolbachia introduction threshold. Our recent findings manifest that the threshold is highly dependent on the initial population size once Wolbachia infection alters the logistic control death rate of infected females. However, counting mosquitoes is beyond the realms of possibility. A plausible method is to monitor the infection frequency. We propose the concept of Wolbachia enhancing domain in which the infection frequency keeps increasing. A detailed description of the domain is presented. Our results suggest that both the initial population size and the infection frequency should be taken into account for optimal release strategies. Both Wolbachia fixation and extinction permit the oscillation of the infection frequency.}, } @article {pmid30021000, year = {2018}, author = {, }, title = {Correction: First detection of Wolbachia in the New Zealand biota.}, journal = {PloS one}, volume = {13}, number = {7}, pages = {e0201151}, pmid = {30021000}, issn = {1932-6203}, abstract = {[This corrects the article DOI: 10.1371/journal.pone.0195517.].}, } @article {pmid30020933, year = {2018}, author = {Moretti, R and Yen, PS and Houé, V and Lampazzi, E and Desiderio, A and Failloux, AB and Calvitti, M}, title = {Combining Wolbachia-induced sterility and virus protection to fight Aedes albopictus-borne viruses.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {7}, pages = {e0006626}, pmid = {30020933}, issn = {1935-2735}, mesh = {Aedes/genetics/*microbiology/*physiology ; Animals ; Breeding ; Chikungunya Fever/transmission/virology ; Chikungunya virus/physiology ; Dengue/transmission/virology ; Dengue Virus/physiology ; Female ; Humans ; Infertility ; Male ; Mosquito Control/*methods ; Mosquito Vectors/genetics/*microbiology/*physiology ; Wolbachia/*physiology ; Zika Virus/physiology ; Zika Virus Infection/transmission/virology ; }, abstract = {Among the strategies targeting vector control, the exploitation of the endosymbiont Wolbachia to produce sterile males and/or invasive females with reduced vector competence seems to be promising. A new Aedes albopictus transinfection (ARwP-M) was generated by introducing wMel Wolbachia in the ARwP line which had been established previously by replacing wAlbA and wAlbB Wolbachia with the wPip strain. Various infection and fitness parameters were studied by comparing ARwP-M, ARwP and wild-type (SANG population) Ae. albopictus sharing the same genetic background. Moreover, the vector competence of ARwP-M related to chikungunya, dengue and zika viruses was evaluated in comparison with ARwP. ARwP-M showed a 100% rate of maternal inheritance of wMel and wPip Wolbachia. Survival, female fecundity and egg fertility did not show to differ between the three Ae. albopictus lines. Crosses between ARwP-M males and SANG females were fully unfertile regardless of male age while egg hatch in reverse crosses increased from 0 to about 17% with SANG males aging from 3 to 17 days. When competing with SANG males for SANG females, ARwP-M males induced a level of sterility significantly higher than that expected for an equal mating competitiveness (mean Fried index of 1.71 instead of 1). The overall Wolbachia density in ARwP-M females was about 15 fold higher than in ARwP, mostly due to the wMel infection. This feature corresponded to a strongly reduced vector competence for chikungunya and dengue viruses (in both cases, 5 and 0% rates of transmission at 14 and 21 days post infection) with respect to ARwP females. Results regarding Zika virus did not highlight significant differences between ARwP-M and ARwP. However, none of the tested ARwP-M females was capable at transmitting ZIKV. These findings are expected to promote the exploitation of Wolbachia to suppress the wild-type Ae. albopictus populations.}, } @article {pmid30006543, year = {2018}, author = {Karatepe, B and Aksoy, S and Karatepe, M}, title = {Investigation of Wolbachia spp. and Spiroplasma spp. in Phlebotomus species by molecular methods.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {10616}, pmid = {30006543}, issn = {2045-2322}, support = {R01 AI051584/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA, Bacterial/isolation & purification ; Phlebotomus/*microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Spiroplasma/genetics/*isolation & purification ; Turkey ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The aim of this study was to determine the presence of Wolbachia spp. and Spiroplasma spp. in natural populations of sand flies in Turkey by molecular methods. A total of 40 Phlebotomus specimens (19 female and 21 male) were used in this study. Genomic DNA from whole sand flies was isolated and Wolbachia spp. infection prevalence was investigated by using Wolbachia gene specific primer sets (wsp and GroEL). In addition, the DNA were analyzed for the presence of Spiroplasma infections utilizing bacterium specific 16 S rDNA PCR-amplification primers. Results of this analysis showed a Wolbachia infection prevalence of 70% (28/40). There was no sex-bias in infection prevalence, being 76% (16/21) and 63% (12/19) in males and females, respectively. Analysis of Spiroplasma infections indicated that 26% (5/19) of female sand flies were positive for infection, while none of the screened males (0/21) were positive. Of the 40 sand fly samples, only 2 were found to be positive for both Wolbachia spp. and Spiroplasma spp. The present study demonstrates the presence of Wolbachia and Spiroplasma infections in the natural sand fly populations in Turkey. This is the first report on Spiroplasma infection in the sand flies from Turkey.}, } @article {pmid29989657, year = {2018}, author = {Guo, Y and Hoffmann, AA and Xu, XQ and Zhang, X and Huang, HJ and Ju, JF and Gong, JT and Hong, XY}, title = {Wolbachia-induced apoptosis associated with increased fecundity in Laodelphax striatellus (Hemiptera: Delphacidae).}, journal = {Insect molecular biology}, volume = {27}, number = {6}, pages = {796-807}, doi = {10.1111/imb.12518}, pmid = {29989657}, issn = {1365-2583}, support = {//Medical Research Council/United Kingdom ; }, mesh = {Amino Acid Sequence ; Animals ; *Apoptosis ; Caspases/genetics/*metabolism ; Female ; Fertility ; Genetic Fitness ; Hemiptera/microbiology/*physiology ; Ovary/microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia influence the fitness of their invertebrate hosts. They have effects on reproductive incompatibility and egg production. Although the former are well characterized, the mechanistic basis of the latter is unclear. Here, we investigate whether apoptosis, which has been implicated in fecundity in model insects, influences the interaction between fecundity and Wolbachia in the planthopper Laodelphax striatellus. Wolbachia-infected females produced about 30% more eggs than uninfected females. We used the terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling staining to visualize apoptosis. Microscopic observations indicated that the Wolbachia strain wStri increased the number of ovarioles that contained apoptotic nurse cells in both young and aged adult females. The frequency of apoptosis was much higher in the infected females. The increased fecundity appeared to be a result of apoptosis of nurse cells, which provide nutrients to the growing oocytes. In addition, cell apoptosis inhibition by caspase messenger RNA interference in Wolbachia-infected L. striatellus markedly decreased egg numbers. Together, these data suggest that wStri might enhance fecundity by increasing the number of apoptotic cells in the ovaries in a caspase-dependent manner. Our findings establish a link between Wolbachia-induced apoptosis and egg production effects mediated by Wolbachia, although the way in which the endosymbiont influences caspase levels remains to be determined.}, } @article {pmid29981933, year = {2018}, author = {Ali, H and Muhammad, A and Bala, NS and Wang, G and Chen, Z and Peng, Z and Hou, Y}, title = {Genomic evaluations of Wolbachia and mtDNA in the population of coconut hispine beetle, Brontispa longissima (Coleoptera: Chrysomelidae).}, journal = {Molecular phylogenetics and evolution}, volume = {127}, number = {}, pages = {1000-1009}, doi = {10.1016/j.ympev.2018.07.003}, pmid = {29981933}, issn = {1095-9513}, mesh = {Animals ; Cocos/*parasitology ; Coleoptera/*genetics/*microbiology ; DNA, Mitochondrial/*genetics ; Asia, Eastern ; *Genomics ; Geography ; Haplotypes/genetics ; Likelihood Functions ; Multilocus Sequence Typing ; Phylogeny ; Polymorphism, Genetic ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia pipientis is a diverse, ubiquitous and most prevalent intracellular bacterial group of alpha-Proteobacteria that is concerned with many biological processes in arthropods. The coconut hispine beetle (CHB), Brontispa longissima (Gestro) is an economically important pest of palm cultivation worldwide. In the present study, we comprehensively surveyed the Wolbachia-infection prevalence and mitochondrial DNA (mtDNA) polymorphism in CHB from five different geographical locations, including China's Mainland and Taiwan, Vietnam, Thailand, Malaysia and Indonesia. A total of 540 sequences were screened in this study through three different genes, i.e., cytochrome oxidase subunit I (COI), Wolbachia outer surface protein (wsp) and multilocus sequencing type (MLST) genes. The COI genetic divergence ranges from 0.08% to 0.67%, and likewise, a significant genetic diversity (π = 0.00082; P = 0.049) was noted within and between all analyzed samples. In the meantime, ten different haplotypes (H) were characterized (haplotype diversity = 0.4379) from 21 different locations, and among them, H6 (46 individuals) have shown a maximum number of population clusters than others. Subsequently, Wolbachia-prevalence results indicated that all tested specimens of CHB were found positive (100%), which suggested that CHB was naturally infected with Wolbachia. Wolbachia sequence results (wsp gene) revealed a high level of nucleotide diversity (π = 0.00047) under Tajima's D test (P = 0.049). Meanwhile, the same trend of nucleotide diversity (π = 0.00041) was observed in Wolbachia concatenated MLST locus. Furthermore, phylogenetic analysis (wsp and concatenated MLST genes) revealed that all collected samples of CHB attributed to same Wolbachia B-supergroup. Our results strongly suggest that Wolbachia bacteria and mtDNA were highly concordant with each other and Wolbachia can affect the genetic structure and diversity within the CHB populations.}, } @article {pmid29971900, year = {2018}, author = {Truitt, AM and Kapun, M and Kaur, R and Miller, WJ}, title = {Wolbachia modifies thermal preference in Drosophila melanogaster.}, journal = {Environmental microbiology}, volume = {21}, number = {9}, pages = {3259-3268}, pmid = {29971900}, issn = {1462-2920}, support = {P 28255/FWF_/Austrian Science Fund FWF/Austria ; 0948041//National Science Foundation/ ; P28255-B22//Austrian Science Fund/ ; }, abstract = {Environmental variation can have profound and direct effects on fitness, fecundity, and host-symbiont interactions. Replication rates of microbes within arthropod hosts, for example, are correlated with incubation temperature but less is known about the influence of host-symbiont dynamics on environmental preference. Hence, we conducted thermal preference (Tp) assays and tested if infection status and genetic variation in endosymbiont bacterium Wolbachia affected temperature choice of Drosophila melanogaster. We demonstrate that isogenic flies infected with Wolbachia preferred lower temperatures compared with uninfected Drosophila. Moreover, Tp varied with respect to three investigated Wolbachia variants (wMel, wMelCS, and wMelPop). While uninfected individuals preferred 24.4°C, we found significant shifts of -1.2°C in wMel- and -4°C in flies infected either with wMelCS or wMelPop. We, therefore, postulate that Wolbachia-associated Tp variation within a host species might represent a behavioural accommodation to host-symbiont interactions and trigger behavioural self-medication and bacterial titre regulation by the host.}, } @article {pmid29968550, year = {2018}, author = {Yeap, HL and Endersby-Harshman, NM and Hoffmann, AA}, title = {The Effect of Nonrandom Mating on Wolbachia Dynamics: Implications for Population Replacement and Sterile Releases in Aedes Mosquitoes.}, journal = {The American journal of tropical medicine and hygiene}, volume = {99}, number = {3}, pages = {608-617}, pmid = {29968550}, issn = {1476-1645}, mesh = {Aedes/*microbiology ; Animals ; Dengue/*prevention & control ; Female ; Male ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are known to cause deviations from random mating and affect sperm competition (SC) in some of their arthropod hosts. Because these effects could influence the effectiveness of Wolbachia in mosquito population replacement and suppression programs, we developed a theoretical framework to investigate them and we collected relevant data for the wMel infection in Aedes aegypti. Using incompatibility patterns as a measure of mating success of infected versus uninfected mosquitoes, we found some evidence that uninfected males sire more offspring than infected males. However, our theoretical framework suggests that this effect is unlikely to hamper Wolbachia invasion and has only minor effects on population suppression programs. Nevertheless, we suggest that mating effects and SC need to be monitored in an ongoing manner in release programs, given the possibility of ongoing selection for altered mating patterns.}, } @article {pmid29967922, year = {2019}, author = {Nesvorna, M and Bittner, V and Hubert, J}, title = {The Mite Tyrophagus putrescentiae Hosts Population-Specific Microbiomes That Respond Weakly to Starvation.}, journal = {Microbial ecology}, volume = {77}, number = {2}, pages = {488-501}, pmid = {29967922}, issn = {1432-184X}, support = {GA15-09038S//Grantová Agentura České Republiky/ ; }, mesh = {Acaridae/*microbiology/*physiology ; Animals ; Bacteria/classification/genetics/*isolation & purification ; Bacterial Physiological Phenomena ; Feeding Behavior ; Host Specificity ; *Microbiota ; }, abstract = {The effect of short-term nutrient deprivation was studied in five populations of the mite Tyrophagus putrescentiae with different microbiomes. The fresh weight, nutrient status, respiration, and population growth of the mites were observed for the five mite population-scale samples. The starvation caused the larvae and nymphs to be eliminated, resulting in a significant increase in the fresh weight of starved adult specimens. Three populations were negatively influenced by starvation, and the starved specimens were characterized by a decrease in nutrient status, respiration, and population growth. One population was not influenced or was slightly influenced by starvation, which had no effect on population growth or nutrient contents but caused a significant decrease in respiration. One population was positively influenced by starvation; the population growth increased in starved specimens, and starvation had no effect on respiration. Although starvation altered the bacterial profiles of the microbiomes, these differences were much smaller than those between the populations. The bacterial profiles of Staphylococcus, Bacillus, Kocuria, Brevibacterium, and unidentified Micrococcaceae and Enterobacteriaceae increased in starved specimens, whereas those of Bartonella and Solitalea-like genera were reduced in the starved mite populations. The profiles of the intracellular symbiont Cardinium decreased in the starved specimens, and the Wolbachia profile changes were dependent on the mite population. In mite populations, when the symbionts were rare, their profiles varied stochastically. Correlations between changes in the profiles of the bacterial taxa and mite fitness parameters, including nutrient status (lipids, proteins, saccharides, and glycogen contents), mite population growth, and respiration, were observed. Although the microbiomes were resistant to the perturbations caused by nutrition deficiency, the responses of the mites differed in terms of their population growth, respiration, and nutrient status.}, } @article {pmid29966368, year = {2018}, author = {Ross, PA and Hoffmann, AA}, title = {Continued Susceptibility of the wMel Wolbachia Infection in Aedes aegypti to Heat Stress Following Field Deployment and Selection.}, journal = {Insects}, volume = {9}, number = {3}, pages = {}, pmid = {29966368}, issn = {2075-4450}, abstract = {Aedes aegypti mosquitoes infected with the wMel strain of Wolbachia are being deployed to control the spread of arboviruses around the world through blockage of viral transmission. Blockage by Wolbachia in some scenarios may be affected by the susceptibility of wMel to cyclical heat stress during mosquito larval development. We therefore evaluated the potential to generate a heat-resistant strain of wMel in Ae. aegypti through artificial laboratory selection and through exposure to field temperatures across multiple generations. To generate an artificially selected strain, wMel-infected females reared under cyclical heat stress were crossed to wMel-infected males reared at 26 &deg;C. The low proportion of larvae that hatched founded the next generation, and this process was repeated for eight generations. The wMel heat-selected strain (wMel-HS) was similar to wMel (unselected) in its ability to induce cytoplasmic incompatibility and restore compatibility when larvae were reared under cyclical heat stress, but wMel-HS adults exhibited reduced Wolbachia densities at 26 &deg;C. To investigate the effects of field exposure, we compared the response of wMel-infected Ae. aegypti collected from Cairns, Australia where the infection has been established for seven years, to a wMel-infected population maintained in the laboratory for approximately 60 generations. Field and laboratory strains of wMel did not differ in their response to cyclical heat stress or in their phenotypic effects at 26 &deg;C. The capacity for the wMel infection in Ae. aegypti to adapt to high temperatures therefore appears limited, and alternative strains may need to be considered for deployment in environments where high temperatures are regularly experienced in mosquito breeding sites.}, } @article {pmid29957332, year = {2018}, author = {Ballesteros, C and Geary, JF and Mackenzie, CD and Geary, TG}, title = {Characterization of Divalent Metal Transporter 1 (DMT1) in Brugia malayi suggests an intestinal-associated pathway for iron absorption.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {8}, number = {2}, pages = {341-349}, pmid = {29957332}, issn = {2211-3207}, mesh = {Animals ; Biological Transport ; Brugia malayi/genetics/growth & development/*metabolism ; Cation Transport Proteins/genetics/*metabolism ; *Host-Parasite Interactions ; Intestines/cytology/physiology ; Iron/*metabolism ; Iron Deficiencies ; Mice ; Microvilli/physiology ; Onchocerca volvulus/genetics/growth & development/metabolism ; Real-Time Polymerase Chain Reaction ; Saccharomyces cerevisiae/genetics/metabolism ; Sequence Alignment ; Wolbachia/metabolism ; }, abstract = {Lymphatic filariasis and onchocerciasis are neglected parasitic diseases which pose a threat to public health in tropical and sub-tropical regions. Strategies for control and elimination of these diseases by mass drug administration (MDA) campaigns are designed to reduce symptoms of onchocerciasis and transmission of both parasites to eventually eliminate the burden on public health. Drugs used for MDA are predominantly microfilaricidal, and prolonged rounds of treatment are required for eradication. Understanding parasite biology is crucial to unravelling the complex processes involved in host-parasite interactions, disease transmission, parasite immune evasion, and the emergence of drug resistance. In nematode biology, large gaps still exist in our understanding of iron metabolism, iron-dependent processes and their regulation. The acquisition of iron from the host is a crucial determinant of the success of a parasitic infection. Here we identify a filarial ortholog of Divalent Metal Transporter 1 (DMT1), a member of a highly conserved family of NRAMP proteins that play an essential role in the transport of ferrous iron in many species. We cloned and expressed the B. malayi NRAMP ortholog in the iron-deficient fet3fet4 strain of Saccharomyces cerevisiae, performed qPCR to estimate stage-specific expression, and localized expression of this gene by immunohistochemistry. Results from functional iron uptake assays showed that expression of this gene in the iron transport-deficient yeast strain significantly rescued growth in low-iron medium. DMT1 was highly expressed in adult female and male B. malayi and Onchocerca volvulus. Immunolocalization revealed that DMT1 is expressed in the intestinal brush border, lateral chords, and reproductive tissues of males and females, areas also inhabited by Wolbachia. We hypothesize based on our results that DMT1 in B. malayi functions as an iron transporter. The presence of this transporter in the intestine supports the hypothesis that iron acquisition by adult females requires oral ingestion and suggests that the intestine plays a functional role in at least some aspects of nutrient uptake.}, } @article {pmid29950416, year = {2018}, author = {Parry, R and Asgari, S}, title = {Aedes Anphevirus: an Insect-Specific Virus Distributed Worldwide in Aedes aegypti Mosquitoes That Has Complex Interplays with Wolbachia and Dengue Virus Infection in Cells.}, journal = {Journal of virology}, volume = {92}, number = {17}, pages = {}, pmid = {29950416}, issn = {1098-5514}, mesh = {Aedes/microbiology/*virology ; Animals ; Cell Line ; Chlorocebus aethiops ; Dengue Virus/physiology ; Evolution, Molecular ; Gene Expression Profiling/*methods ; Genome, Viral ; Host Specificity ; Humans ; Infectious Disease Transmission, Vertical/veterinary ; Insect Viruses/classification/physiology ; Mononegavirales/classification/*physiology ; Mosquito Vectors/microbiology/virology ; Phylogeny ; Sequence Analysis, RNA ; Vero Cells ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Insect-specific viruses (ISVs) of the yellow fever mosquito Aedes aegypti have been demonstrated to modulate transmission of arboviruses such as dengue virus (DENV) and West Nile virus by the mosquito. The diversity and composition of the virome of A. aegypti, however, remains poorly understood. In this study, we characterized Aedes anphevirus (AeAV), a negative-sense RNA virus from the order Mononegavirales AeAV identified from Aedes cell lines was infectious to both A. aegypti and Aedes albopictus cells but not to three mammalian cell lines. To understand the incidence and genetic diversity of AeAV, we assembled 17 coding-complete and two partial genomes of AeAV from available transcriptome sequencing (RNA-Seq) data. AeAV appears to transmit vertically and be present in laboratory colonies, wild-caught mosquitoes, and cell lines worldwide. Phylogenetic analysis of AeAV strains indicates that as the A. aegypti mosquito has expanded into the Americas and Asia-Pacific, AeAV has evolved into monophyletic African, American, and Asia-Pacific lineages. The endosymbiotic bacterium Wolbachia pipientis restricts positive-sense RNA viruses in A. aegypti Reanalysis of a small RNA library of A. aegypti cells coinfected with AeAV and Wolbachia produces an abundant RNA interference (RNAi) response consistent with persistent virus replication. We found Wolbachia enhances replication of AeAV compared to a tetracycline-cleared cell line, and AeAV modestly reduces DENV replication in vitro The results from our study improve understanding of the diversity and evolution of the virome of A. aegypti and adds to previous evidence that shows Wolbachia does not restrict a range of negative-strand RNA viruses.IMPORTANCE The mosquito Aedes aegypti transmits a number of arthropod-borne viruses (arboviruses), such as dengue virus and Zika virus. Mosquitoes also harbor insect-specific viruses that may affect replication of pathogenic arboviruses in their body. Currently, however, there are only a few insect-specific viruses described from A. aegypti in the literature. Here, we characterize a novel negative-strand virus, AeAV. Meta-analysis of A. aegypti samples showed that it is present in A. aegypti mosquitoes worldwide and is vertically transmitted. Wolbachia-transinfected mosquitoes are currently being used in biocontrol, as they effectively block transmission of several positive-sense RNA viruses in mosquitoes. Our results demonstrate that Wolbachia enhances the replication of AeAV and modestly reduces dengue virus replication in a cell line model. This study expands our understanding of the virome in A. aegypti as well as providing insight into the complexity of the Wolbachia virus restriction phenotype.}, } @article {pmid29947761, year = {2018}, author = {Faria, VG and Martins, NE and Schlötterer, C and Sucena, É}, title = {Readapting to DCV Infection without Wolbachia: Frequency Changes of Drosophila Antiviral Alleles Can Replace Endosymbiont Protection.}, journal = {Genome biology and evolution}, volume = {10}, number = {7}, pages = {1783-1791}, pmid = {29947761}, issn = {1759-6653}, support = {P 27630/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Adaptation, Physiological ; Alleles ; Animals ; Drosophila melanogaster/genetics/*microbiology/physiology/*virology ; Genome, Insect ; Host-Pathogen Interactions ; Insect Viruses/*physiology ; Polymorphism, Genetic ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {There is now ample evidence that endosymbionts can contribute to host adaptation to environmental challenges. However, how endosymbiont presence affects the adaptive trajectory and outcome of the host is yet largely unexplored. In Drosophila, Wolbachia confers protection to RNA virus infection, an effect that differs between Wolbachia strains and can be targeted by selection. Adaptation to RNA virus infections is mediated by both Wolbachia and the host, raising the question of whether adaptive genetic changes in the host vary with the presence/absence of the endosymbiont. Here, we address this question using a polymorphic D. melanogaster population previously adapted to DCV infection for 35 generations in the presence of Wolbachia, from which we removed the endosymbiont and followed survival over the subsequent 20 generations of infection. After an initial severe drop, survival frequencies upon DCV selection increased significantly, as seen before in the presence of Wolbachia. Whole-genome sequencing, revealed that the major genes involved in the first selection experiment, pastrel and Ubc-E2H, continued to be selected in Wolbachia-free D. melanogaster, with the frequencies of protective alleles being closer to fixation in the absence of Wolbachia. Our results suggest that heterogeneity in Wolbachia infection status may be sufficient to maintain polymorphisms even in the absence of costs.}, } @article {pmid29938064, year = {2018}, author = {Amuzu, HE and Tsyganov, K and Koh, C and Herbert, RI and Powell, DR and McGraw, EA}, title = {Wolbachia enhances insect-specific flavivirus infection in Aedes aegypti mosquitoes.}, journal = {Ecology and evolution}, volume = {8}, number = {11}, pages = {5441-5454}, pmid = {29938064}, issn = {2045-7758}, abstract = {Mosquitoes transmit a diverse group of human flaviviruses including West Nile, dengue, yellow fever, and Zika viruses. Mosquitoes are also naturally infected with insect-specific flaviviruses (ISFs), a subgroup of the family not capable of infecting vertebrates. Although ISFs are not medically important, they are capable of altering the mosquito's susceptibility to flaviviruses and may alter host fitness. Wolbachia is an endosymbiotic bacterium of insects that when present in mosquitoes limits the replication of co-infecting pathogens, including flaviviruses. Artificially created Wolbachia-infected Aedes aegypti mosquitoes are being released into the wild in a series of trials around the globe with the hope of interrupting dengue and Zika virus transmission from mosquitoes to humans. Our work investigated the effect of Wolbachia on ISF infection in wild-caught Ae. aegypti mosquitoes from field release zones. All field mosquitoes were screened for the presence of ISFs using general degenerate flavivirus primers and their PCR amplicons sequenced. ISFs were found to be common and widely distributed in Ae. aegypti populations. Field mosquitoes consistently had higher ISF infection rates and viral loads compared to laboratory colony material indicating that environmental conditions may modulate ISF infection in Ae. aegypti. Surprisingly, higher ISF infection rates and loads were found in Wolbachia-infected mosquitoes compared to the Wolbachia-free mosquitoes. Our findings demonstrate that the symbiont is capable of manipulating the mosquito virome and that Wolbachia-mediated viral inhibition is not universal for flaviviruses. This may have implications for the Wolbachia-based DENV control strategy if ISFs confer fitness effects or alter mosquito susceptibility to other flaviviruses.}, } @article {pmid29931623, year = {2019}, author = {Arai, H and Hirano, T and Akizuki, N and Abe, A and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Multiple Infection and Reproductive Manipulations of Wolbachia in Homona magnanima (Lepidoptera: Tortricidae).}, journal = {Microbial ecology}, volume = {77}, number = {1}, pages = {257-266}, pmid = {29931623}, issn = {1432-184X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Bacterial Physiological Phenomena ; Cytoplasm ; DNA, Bacterial/analysis ; Female ; Larva/microbiology ; Male ; Moths/drug effects/*genetics/*microbiology ; Phenotype ; Pupa/microbiology ; Reproduction/*physiology ; Sex Ratio ; Symbiosis ; *Wolbachia/classification/genetics/physiology ; }, abstract = {Endosymbiotic bacterium Wolbachia interacts with host in either a mutualistic or parasitic manner. Wolbachia is frequently identified in various arthropod species, and to date, Wolbachia infections have been detected in different insects. Here, we found a triple Wolbachia infection in Homona magnanima, a serious tea pest, and investigated the effects of three infecting Wolbachia strains (wHm-a, -b, and -c) on the host. Starting with the triple-infected host line (W[abc]), which was collected in western Tokyo in 1999 and maintained in laboratory, we established an uninfected line (W[-]) and three singly infected lines (W[a], W[b], and W[c]) using antibiotics. Mating experiments with the host lines revealed that only wHm-b induced cytoplasmic incompatibility (CI) in H. magnanima, with the intensities of CI different between the W[b] and W[abc] lines. Regarding mutualistic effects, wHm-c shortened larval development time and increased pupal weight in both the W[c] and W[abc] lines to the same extent, whereas no distinct phenotype was observed in lines singly infected with wHm-a. Based on quantitative PCR analysis, Wolbachia density in the W[a] line was higher than in the other host lines (p < 0.01, n = 10). Wolbachia density in the W[b] line was also higher than in the W[c] and W[abc] lines, while no difference was observed between the W[c] and W[abc] lines. These results indicate that the difference in the CI intensity between a single or multiple infection may be attributed to the difference in wHm-b density. However, no correlation was observed between mutualistic effects and Wolbachia density.}, } @article {pmid29908171, year = {2018}, author = {Goindin, D and Cannet, A and Delannay, C and Ramdini, C and Gustave, J and Atyame, C and Vega-Rúa, A}, title = {Screening of natural Wolbachia infection in Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus from Guadeloupe (French West Indies).}, journal = {Acta tropica}, volume = {185}, number = {}, pages = {314-317}, doi = {10.1016/j.actatropica.2018.06.011}, pmid = {29908171}, issn = {1873-6254}, mesh = {Aedes/*microbiology ; Animals ; Culex/*microbiology ; Guadeloupe ; Mosquito Control/methods ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Guadeloupe islands are threatened by several mosquito-borne viruses such as Dengue, Chikungunya, Zika and West Nile virus. It appears essential to look for alternative mosquito control methods such as the incompatible insect technique (ITT) aiming at sterilizing wild females by inundative releases of incompatible males. Before considering the implementation of such a strategy, the characterization of genetic diversity of the endocellular bacterium Wolbachia regarding the local mosquito populations is a critical issue. Here, for the first time, we describe the prevalence and diversity of Wolbachia in natural populations of three mosquito species from Guadeloupe: Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus. The detection of Wolbachia in natural Ae. aegypti, Ae. taeniorhynchus and Cx. quinquefasciatus populations was conducted by studying Wolbachia 16S ribosomal RNA gene using a TaqMan quantitative real-time PCR and results were confirmed by conventional PCR and sequencing. In addition, molecular typing of wPip strains in Cx. quinquefasciatus was done by PCR-RFLP. We did not find Wolbachia infection in any of Ae. aegypti and Ae. taeniorhynchus studied populations. Natural Wolbachia infection was detected in Cx. quinquefasciatus with prevalence varying from 79.2% to 95.8%. In addition, no polymorphism was found between the Wolbachia strains infecting Cx. quinquefasciatus specimens, all carrying an infection from the same Wolbachia genetic wPip-I group. These results pave the way for the evaluation of the feasibility of IIT programs to fight against these medically-important mosquito species in Guadeloupe.}, } @article {pmid29901734, year = {2018}, author = {Gloria-Soria, A and Chiodo, TG and Powell, JR}, title = {Lack of Evidence for Natural Wolbachia Infections in Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {55}, number = {5}, pages = {1354-1356}, pmid = {29901734}, issn = {1938-2928}, support = {R01 AI101112/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; *Wolbachia ; }, abstract = {Wolbachia is a genus of endosymbiotic bacteria that infects 66% of all insect species. Its major impact on insects is in reproduction: sterility, production of one sex, and/or parthenogenesis. Another effect was discovered when the disease-transmitting mosquito, Aedes aegypti Linnaeus (Diptera: Culicidae), was infected with Wolbachia isolated from Drosophila: infected female mosquitoes became less capable of transmitting diseases such as dengue fever and chikungunya. This has led to releases of Ae. aegypti carrying Wolbachia in an attempt to control disease. An open question is whether there are natural Wolbachia infections of this mosquito. We assayed DNA from 2,663 Ae. aegypti from 27 countries on six continents, 230 from laboratory strains, and 72 Aedes mascarensis MacGregor (Diptera: Culicidae) for presence of Wolbachia DNA. Within the limits of our polymerase chain reaction-based assay, we found no evidence of Wolbachia, suggesting that natural infections of this endosymbiont are unlikely to occur throughout the worldwide distribution of Ae. aegypti.}, } @article {pmid29897678, year = {2019}, author = {Uribe-Alvarez, C and Chiquete-Félix, N and Morales-García, L and Bohórquez-Hernández, A and Delgado-Buenrostro, NL and Vaca, L and Peña, A and Uribe-Carvajal, S}, title = {Wolbachia pipientis grows in Saccharomyces cerevisiae evoking early death of the host and deregulation of mitochondrial metabolism.}, journal = {MicrobiologyOpen}, volume = {8}, number = {4}, pages = {e00675}, pmid = {29897678}, issn = {2045-8827}, mesh = {Animals ; Bacterial Proteins/genetics/metabolism ; Host-Pathogen Interactions ; Insecta/*microbiology/physiology ; Mitochondria/*metabolism ; Oxidative Phosphorylation ; Saccharomyces cerevisiae/chemistry/*metabolism ; Wolbachia/*growth & development ; }, abstract = {Wolbachia sp. has colonized over 70% of insect species, successfully manipulating host fertility, protein expression, lifespan, and metabolism. Understanding and engineering the biochemistry and physiology of Wolbachia holds great promise for insect vector-borne disease eradication. Wolbachia is cultured in cell lines, which have long duplication times and are difficult to manipulate and study. The yeast strain Saccharomyces cerevisiae W303 was used successfully as an artificial host for Wolbachia wAlbB. As compared to controls, infected yeast lost viability early, probably as a result of an abnormally high mitochondrial oxidative phosphorylation activity observed at late stages of growth. No respiratory chain proteins from Wolbachia were detected, while several Wolbachia F1 F0 -ATPase subunits were revealed. After 5 days outside the cell, Wolbachia remained fully infective against insect cells.}, } @article {pmid29893631, year = {2018}, author = {Vasconcelos, EJR and Billeter, SA and Jett, LA and Meinersmann, RJ and Barr, MC and Diniz, PPVP and Oakley, BB}, title = {Assessing Cat Flea Microbiomes in Northern and Southern California by 16S rRNA Next-Generation Sequencing.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {18}, number = {9}, pages = {491-499}, doi = {10.1089/vbz.2018.2282}, pmid = {29893631}, issn = {1557-7759}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; California/epidemiology ; Cat Diseases/epidemiology/parasitology ; Cats ; Ctenocephalides/*microbiology ; DNA, Bacterial/genetics ; RNA, Bacterial/*genetics ; RNA, Ribosomal, 16S/*genetics ; }, abstract = {Flea-borne diseases (FBDs) impact both human and animal health worldwide. Because adult fleas are obligately hematophagous and can harbor potential pathogens, fleas act as ectoparasites of vertebrates, as well as zoonotic disease vectors. Cat fleas (Ctenocephalides felis) are important vectors of two zoonotic bacterial genera listed as priority pathogens by the National Institute of Allergy and Infectious Diseases (NIAID-USA): Bartonella spp. and Rickettsia spp., causative agents of bartonelloses and rickettsioses, respectively. In this study, we introduce the first microbiome analysis of C. felis samples from California, determining the presence and abundance of relevant pathogenic genera by characterizing the cat flea microbiome through 16S rRNA next-generation sequencing (16S-NGS). Samples from both northern (NoCal) and southern (SoCal) California were assessed to expand current knowledge regarding FBDs in the state. We identified Rickettsia and Bartonella, as well as the endosymbiont Wolbachia, as the most abundant genera, followed by less abundant taxa. In comparison to our previous study screening Californian cat fleas for rickettsiae using PCR/digestion/sequencing of the ompB gene, the 16S-NGS approach applied herein showed a 95% level of agreement in detecting Rickettsia spp. There was no overall difference in microbiome diversity between NoCal and SoCal samples. Bacterial taxa identified by 16S-NGS in this study may help to improve epidemiological investigations, pathogen surveillance efforts, and clinical diagnostics of FBDs in California and elsewhere.}, } @article {pmid29891919, year = {2018}, author = {Balvín, O and Roth, S and Talbot, B and Reinhardt, K}, title = {Co-speciation in bedbug Wolbachia parallel the pattern in nematode hosts.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {8797}, pmid = {29891919}, issn = {2045-2322}, mesh = {Animals ; Bedbugs/*microbiology ; Biosynthetic Pathways/genetics ; Biotin/biosynthesis ; *Genetic Speciation ; *Symbiosis ; Wolbachia/*classification/genetics/*isolation & purification/metabolism ; }, abstract = {Wolbachia bacteria, vertically transmitted intracellular endosymbionts, are associated with two major host taxa in which they show strikingly different symbiotic modes. In some taxa of filarial nematodes, where Wolbachia are strictly obligately beneficial to the host, they show complete within- and among-species prevalence as well as co-phylogeny with their hosts. In arthropods, Wolbachia usually are parasitic; if beneficial effects occurs, they can be facultative or obligate, related to host reproduction. In arthropods, the prevalence of Wolbachia varies within and among taxa, and no co-speciation events are known. However, one arthropod species, the common bedbug Cimex lectularius was recently found to be dependent on the provision of biotin and riboflavin by Wolbachia, representing a unique case of Wolbachia providing nutritional and obligate benefits to an arthropod host, perhaps even in a mutualistic manner. Using the presence of presumably functional biotin gene copies, our study demonstrates that the obligate relationship is maintained at least in 10 out of 15 species of the genera Cimex and Paracimex. The remaining five species harboured Wolbachia as well, demonstrating the first known case of 100% prevalence of Wolbachia among higher arthropod taxa. Moreover, we show the predicted co-cladogenesis between Wolbachia and their bedbug hosts, also as the first described case of Wolbachia co-speciation in arthropods.}, } @article {pmid29890648, year = {2018}, author = {Badawi, M and Moumen, B and Giraud, I and Grève, P and Cordaux, R}, title = {Investigating the Molecular Genetic Basis of Cytoplasmic Sex Determination Caused by Wolbachia Endosymbionts in Terrestrial Isopods.}, journal = {Genes}, volume = {9}, number = {6}, pages = {}, pmid = {29890648}, issn = {2073-4425}, abstract = {In animals, sexual differences between males and females are usually determined by sex chromosomes. Alternatively, sex may also be determined by vertically transmitted intracellular microbial endosymbionts. The best known cytoplasmic sex manipulative endosymbiont is Wolbachia which can, for instance, feminize genetic males into phenotypic females in the terrestrial isopod Armadillidium vulgare. However, the molecular genetic basis of cytoplasmic sex determination is unknown. To identify candidate genes of feminization induced by Wolbachia strain wVulC from A. vulgare, we sequenced the genome of Wolbachia strain wCon from Cylisticus convexus, the most closely related known Wolbachia strain to wVulC that does not induce feminization, and compared it to the wVulC genome. Then, we performed gene expression profiling of the 216 resulting wVulC candidate genes throughout host developmental stages in A. vulgare and the heterologous host C. convexus. We identified a set of 35 feminization candidate genes showing differential expression during host sexual development. Interestingly, 27 of the 35 genes are present in the f element, which is a piece of a feminizing Wolbachia genome horizontally transferred into the nuclear genome of A. vulgare and involved in female sex determination. Assuming that the molecular genetic basis of feminization by Wolbachia and the f element is the same, the 27 genes are candidates for acting as master sex determination genes in A. vulgare females carrying the f element.}, } @article {pmid29876068, year = {2018}, author = {Zhang, L and Yun, Y and Hu, G and Peng, Y}, title = {Insights into the bacterial symbiont diversity in spiders.}, journal = {Ecology and evolution}, volume = {8}, number = {10}, pages = {4899-4906}, pmid = {29876068}, issn = {2045-7758}, abstract = {Most spiders are natural enemies of pests, and it is beneficial for the biological control of pests to learn the relationships between symbionts and their spider hosts. Research on the bacterial communities of insects has been conducted recently, but only a few studies have addressed the bacterial communities of spiders. To obtain a complete overview of the microbial communities of spiders, we examined eight species of spider (Pirata subpiraticus, Agelena difficilis, Artema atlanta, Nurscia albofasciata, Agelena labyrinthica, Ummeliata insecticeps, Dictis striatipes, and Hylyphantes graminicola) with high-throughput sequencing based on the V3 and V4 regions of the 16S rRNA gene. The bacterial communities of the spider samples were dominated by five types of endosymbionts, Wolbachia, Cardinium, Rickettsia, Spiroplasma, and Rickettsiella. The dominant OTUs (operational taxonomic units) from each of the five endosymbionts were analyzed, and the results showed that different spider species were usually dominated by special OTUs. In addition to endosymbionts, Pseudomonas, Sphingomonas, Acinetobacter, Novosphingobium, Aquabacterium, Methylobacterium, Brevundimonas, Rhizobium, Bradyrhizobium, Citrobacter, Arthrobacter, Pseudonocardia, Microbacterium, Lactobacillus, and Lactococcus were detected in spider samples in our study. Moreover, the abundance of Sphingomonas, Methylobacterium, Brevundimonas, and Rhizobium in the spider D. striatipes was significantly higher (p < .05) than the bacterial abundance of these species in seven other spider species. These findings suggest that same as in insects, co-infection of multiple types of endosymbionts is common in the hosts of the Araneae order, and other bacterial taxa also exist in spiders besides the endosymbionts.}, } @article {pmid29868291, year = {2018}, author = {Li, J and Wang, N and Liu, Y and Qiu, S}, title = {Proteomics of Nasonia vitripennis and the effects of native Wolbachia infection on N. vitripennis.}, journal = {PeerJ}, volume = {6}, number = {}, pages = {e4905}, pmid = {29868291}, issn = {2167-8359}, abstract = {BACKGROUND: Nasonia vitripennis, a parasitic wasp, is a good model organism to study developmental and evolutionary genetics and to evaluate the interactions between insect hosts and their symbionts. Wolbachia may be the most prevalent endosymbiont among insect species due to their special ability to improve the fitness of the infected hosts. Transinfection of bacteria or fungi could substantially alter the expression of host immune system components. However, few studies have focused on the effects of native Wolbachia infection. Accordingly, in this study, we evaluated the proteomics of N. vitripennis following Wolbachia infection.

METHODS: We studied the proteomics of N. vitripennis following native Wolbachia infection and in antibiotic-treated Wolbachia-free samples using isobaric tags for relative and absolute quantification-liquid chromatography tandem mass spectrometry, accompanying with some ecological experiments.

RESULTS: In total, 3,096 proteins were found to be associated with a wide range of biological processes, molecular functions, and cellular components. Interestingly, there were few significant changes in immune or reproductive proteins between samples with and without Wolbachia infection. Differentially expressed proteins were involved in the binding process, catalytic activity, and the metabolic process, as confirmed by quantitative reverse transcription polymerase chain reaction.

DISCUSSION: Invasion of any pathogen or bacterium within a short time can cause an immunoreaction in the host. Our results implied that during the long process of coexistence, the immune system of the host was not as sensitive as when the symbiont initially infected the host, implying that the organisms had gradually adjusted to cohabitation.}, } @article {pmid29864164, year = {2018}, author = {Palmer, WH and Medd, NC and Beard, PM and Obbard, DJ}, title = {Isolation of a natural DNA virus of Drosophila melanogaster, and characterisation of host resistance and immune responses.}, journal = {PLoS pathogens}, volume = {14}, number = {6}, pages = {e1007050}, pmid = {29864164}, issn = {1553-7374}, support = {WT085064/WT_/Wellcome Trust/United Kingdom ; WT104915MA/WT_/Wellcome Trust/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; BB/J004324/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; WT095831/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Biological Evolution ; DNA Virus Infections/genetics/immunology/*virology ; DNA Viruses/*isolation & purification/physiology ; Drosophila melanogaster/growth & development/*immunology/microbiology/virology ; Female ; Genetic Variation ; Host-Pathogen Interactions/*immunology ; Male ; Symbiosis ; Viral Proteins/*genetics ; Virus Replication ; Wolbachia/*growth & development/immunology/isolation & purification ; }, abstract = {Drosophila melanogaster has played a key role in our understanding of invertebrate immunity. However, both functional and evolutionary studies of host-virus interaction in Drosophila have been limited by a dearth of native virus isolates. In particular, despite a long history of virus research, DNA viruses of D. melanogaster have only recently been described, and none have been available for experimental study. Here we report the isolation and comprehensive characterisation of Kallithea virus, a large double-stranded DNA virus, and the first DNA virus to have been reported from wild populations of D. melanogaster. We find that Kallithea virus infection is costly for adult flies, reaching high titres in both sexes and disproportionately reducing survival in males, and movement and late fecundity in females. Using the Drosophila Genetic Reference Panel, we quantify host genetic variance for virus-induced mortality and viral titre and identify candidate host genes that may underlie this variation, including Cdc42-interacting protein 4. Using full transcriptome sequencing of infected males and females, we examine the transcriptional response of flies to Kallithea virus infection and describe differential regulation of virus-responsive genes. This work establishes Kallithea virus as a new tractable model to study the natural interaction between D. melanogaster and DNA viruses, and we hope it will serve as a basis for future studies of immune responses to DNA viruses in insects.}, } @article {pmid29859884, year = {2018}, author = {Hornok, S and Horváth, G and Takács, N and Farkas, R and Szőke, K and Kontschán, J}, title = {Molecular evidence of a badger-associated Ehrlichia sp., a Candidatus Neoehrlichia lotoris-like genotype and Anaplasma marginale in dogs.}, journal = {Ticks and tick-borne diseases}, volume = {9}, number = {5}, pages = {1302-1309}, doi = {10.1016/j.ttbdis.2018.05.012}, pmid = {29859884}, issn = {1877-9603}, mesh = {Anaplasma marginale/genetics/isolation & purification ; Anaplasma phagocytophilum/genetics/isolation & purification ; Anaplasmataceae/*genetics/isolation & purification ; Anaplasmataceae Infections/blood/microbiology/*veterinary ; Animals ; DNA, Bacterial/blood/genetics ; Dogs ; Ehrlichia/*genetics/isolation & purification ; Genotype ; Ixodes/*microbiology ; Male ; Mustelidae/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/genetics/isolation & purification ; }, abstract = {The family Anaplasmataceae contains pathogenic and endosymbiotic bacteria of veterinary-medical importance. In this study, 90 blood samples from rural dogs, five blood samples from road-killed European badgers and 34 ticks, i.e. 27 Ixodes (Pholeoixodes) canisuga, six I. (Ph.) hexagonus and one Haemaphysalis concinna collected from the badgers were molecularly analysed for members of Anaplasmataceae. Apart from the molecular evidence of Anaplasma phagocytophilum in one dog and Wolbachia sp. associated with Dirofilaria repens in five dogs, four species/genotypes not yet known to occur in canine hosts have also been found. These included A. marginale in two dogs, a badger-associated Ehrlichia sp. in one dog, a Candidatus Neoehrlichia lotoris-like genotype in six dogs and the DNA of arthropod-associated wolbachiae in three dogs. In two badgers the DNA from the Candidatus N. lotoris-like genotype was identified. Among ticks, four I. canisuga carried the DNA of the above badger-associated Ehrlichia sp., one I. canisuga contained the Candidatus N. lotoris-like genotype, and in H. concinna Wolbachia DNA was present. In conclusion, results shown here should be interpreted as the first molecular evidence for exposure of dogs to three members of Anaplasmataceae, i.e. A. marginale, a badger-associated Ehrlichia sp. and a Candidatus N. lotoris-like agent. The presence of DNA in the blood of relevant animals may also indicate susceptibility to these bacteria, but in support of this, further studies are needed.}, } @article {pmid29855331, year = {2018}, author = {Anders, KL and Indriani, C and Ahmad, RA and Tantowijoyo, W and Arguni, E and Andari, B and Jewell, NP and Rances, E and O'Neill, SL and Simmons, CP and Utarini, A}, title = {The AWED trial (Applying Wolbachia to Eliminate Dengue) to assess the efficacy of Wolbachia-infected mosquito deployments to reduce dengue incidence in Yogyakarta, Indonesia: study protocol for a cluster randomised controlled trial.}, journal = {Trials}, volume = {19}, number = {1}, pages = {302}, pmid = {29855331}, issn = {1745-6215}, support = {R56 AI134724/AI/NIAID NIH HHS/United States ; }, mesh = {Adolescent ; Adult ; Animals ; Child ; Child, Preschool ; Culicidae/*microbiology ; Dengue/epidemiology/*prevention & control/transmission/virology ; Dengue Virus/*pathogenicity ; Female ; Humans ; Incidence ; Indonesia/epidemiology ; Male ; Middle Aged ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods ; Time Factors ; Wolbachia/*physiology ; Young Adult ; }, abstract = {BACKGROUND: Dengue and other arboviruses transmitted by Aedes aegypti mosquitoes, including Zika and chikungunya, present an increasing public health challenge in tropical regions. Current vector control strategies have failed to curb disease transmission, but continue to be employed despite the absence of robust evidence for their effectiveness or optimal implementation. The World Mosquito Program has developed a novel approach to arbovirus control using Ae. aegypti stably transfected with Wolbachia bacterium, with a significantly reduced ability to transmit dengue, Zika and chikungunya in laboratory experiments. Modelling predicts this will translate to local elimination of dengue in most epidemiological settings. This study protocol describes the first trial to measure the efficacy of Wolbachia in reducing dengue virus transmission in the field.

METHODS/DESIGN: The study is a parallel, two-arm, non-blinded cluster randomised controlled trial conducted in a single site in Yogyakarta, Indonesia. The aim is to determine whether large-scale deployment of Wolbachia-infected Ae. aegypti mosquitoes leads to a measurable reduction in dengue incidence in treated versus untreated areas. The primary endpoint is symptomatic, virologically confirmed dengue virus infection of any severity. The 26 km[2] study area was subdivided into 24 contiguous clusters, allocated randomly 1:1 to receive Wolbachia deployments or no intervention. We use a novel epidemiological study design, the cluster-randomised test-negative design trial, in which dengue cases and arbovirus-negative controls are sampled concurrently from among febrile patients presenting to a network of primary care clinics, with case or control status classified retrospectively based on the results of laboratory diagnostic testing. Efficacy is estimated from the odds ratio of Wolbachia exposure distribution (probability of living in a Wolbachia-treated area) among virologically confirmed dengue cases compared to test-negative controls. A secondary per-protocol analysis allows for individual Wolbachia exposure levels to be assessed to account for movements outside the cluster and the heterogeneity in local Wolbachia prevalence among treated clusters.

DISCUSSION: The findings from this study will provide the first experimental evidence for the efficacy of Wolbachia in reducing dengue incidence. Together with observational evidence that is accumulating from pragmatic deployments of Wolbachia in other field sites, this will provide valuable data to estimate the effectiveness of this novel approach to arbovirus control, inform future cost-effectiveness estimates, and guide plans for large-scale deployments in other endemic settings.

TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT03055585 . Registered on 14 February 2017.}, } @article {pmid29851312, year = {2018}, author = {Dahmani, M and Tahir, D and Cabre, O and Raoult, D and Fenollar, F and Davoust, B and Mediannikov, O}, title = {Prevalence of Anaplasmataceae and Filariidae species in unowned and military dogs in New Caledonia.}, journal = {Veterinary medicine and science}, volume = {4}, number = {2}, pages = {140-149}, pmid = {29851312}, issn = {2053-1095}, mesh = {Anaplasmataceae/physiology ; Anaplasmataceae Infections/epidemiology/parasitology/*veterinary ; Animals ; Dog Diseases/*epidemiology/parasitology ; Dogs ; Female ; Filariasis/epidemiology/parasitology/*veterinary ; Filarioidea/physiology ; Male ; Military Personnel ; New Caledonia/epidemiology ; Ownership ; Prevalence ; }, abstract = {Dogs are competent reservoir hosts of several zoonotic agents, including Filariidae nematodes and Anaplasmataceae family bacteria. The latter family unites human and veterinary pathogens (Anaplasma, Ehrlichia and Neorickettsia bacteria) with Wolbachia, some of which are obligatory endosymbionts of pathogenic filarial nematodes. The epidemiology of Anaplasmataceae and Filariidae species infecting dogs living in kennels in New Caledonia was studied. 64 EDTA blood samples were screened for the presence of Anaplasmataceae and filarial nematodes. Molecular study was conducted using primers and probe targeting the of 23S rRNA long fragment of Anaplasmataceae species. Next, all blood sample was screened for the presence of Filariidae species targeting the primers and probe targeting the COI gene, as well as primers targeting the COI and 5S rRNA genes of all filarial worms. Anaplasma platys was identified in 8/64 (12.5, 95% confidence interval [CI]: 4.4-20.6%) and Wolbachia endosymbiont of Dirofilaria immitis in 8/64 (12.5%, CI: 4.4-20.6%). Filariidae species investigation was performed and showed that 11/64 (17.2%, CI: 7.9-26.4%) dogs were infected with D. immitis, whereas, 2/64 (3.1%, CI: 0.0-7.3%) were infected with Acanthocheilonema reconditum. Finally, we checked the occurrence of co-infection between Anaplasmataceae and Filariidae species. Co-occurrence with Wolbachia endosymbiont of D. immitis was observed in seven dogs, one dog was co-infected with A. platys and A. reconditum and another was co-infected with Wolbachia endosymbiont of D. immitis and A. reconditum. These results are the first report of Anaplasmataceae and Filariidae occurring in dogs in New Caledonia.}, } @article {pmid29851149, year = {2018}, author = {Ote, M and Yamamoto, D}, title = {The Wolbachia protein TomO interacts with a host RNA to induce polarization defects in Drosophila oocytes.}, journal = {Archives of insect biochemistry and physiology}, volume = {99}, number = {1}, pages = {e21475}, doi = {10.1002/arch.21475}, pmid = {29851149}, issn = {1520-6327}, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Body Patterning ; Drosophila simulans/*embryology/*microbiology ; Embryo, Nonmammalian/microbiology ; Embryonic Development ; Oocytes/growth & development ; RNA/metabolism ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an endosymbiont prevalent in arthropods. To maximize its transmission thorough the female germline, Wolbachia induces in infected hosts male-to-female transformation, male killing, parthenogenesis, and cytoplasmic incompatibility, depending on the host species and Wolbachia strain involved. However, the molecular mechanisms underlying these host manipulations by Wolbachia remain largely unknown. The Wolbachia strain wMel, an inhabitant of Drosophila melanogaster, impairs host oogenesis only when transplanted into a heterologous host, for example, Drosophila simulans. We found that egg polarity defects induced by wMel infection in D. simulans can be recapitulated in the natural host D. melanogaster by transgenic overexpression of a variant of the Wolbachia protein Toxic manipulator of oogenesis (TomO), TomOwMel[∆HS] , in the female germline. RNA immunoprecipitation assays demonstrated that TomO physically associates with orb mRNA, which, as a result, fails to interact with the translation repressor Cup. This leads to precocious translation of Orb, a posterior determinant, and thereby to the misspecification of oocytes and accompanying polarity defects. We propose that the ability of TomO to bind to orb mRNA might provide a means for Wolbachia to enter the oocyte located at the posterior end of the egg chamber, thereby accomplishing secure maternal transmission thorough the female germline.}, } @article {pmid29845544, year = {2018}, author = {O'Neill, SL}, title = {The Use of Wolbachia by the World Mosquito Program to Interrupt Transmission of Aedes aegypti Transmitted Viruses.}, journal = {Advances in experimental medicine and biology}, volume = {1062}, number = {}, pages = {355-360}, doi = {10.1007/978-981-10-8727-1_24}, pmid = {29845544}, issn = {0065-2598}, mesh = {Aedes/*microbiology/virology ; Animals ; Australia ; Humans ; Mosquito Control ; Mosquito Vectors/*microbiology/virology ; Virus Diseases/transmission/virology ; *Virus Physiological Phenomena ; Viruses/genetics ; Wolbachia/*physiology ; }, abstract = {The biological control of mosquito transmission by the use of the naturally occurring insect-specific bacterial endosymbiont Wolbachia has been successfully tested in small field trials. The approach has been translated successfully to larger field sites in Townsville, Australia and expanded to more than 10 countries through the Eliminate Dengue Program. The broader application of the program beyond limiting the transmission of dengue and including other Aedes aegypti borne mosquitoes has seen the program growing into a global not-for-profit initiative to be known as the World Mosquito Program.}, } @article {pmid29843598, year = {2018}, author = {Perry, KD and Baker, GJ and Powis, KJ and Kent, JK and Ward, CM and Baxter, SW}, title = {Cryptic Plutella species show deep divergence despite the capacity to hybridize.}, journal = {BMC evolutionary biology}, volume = {18}, number = {1}, pages = {77}, pmid = {29843598}, issn = {1471-2148}, support = {UA00146//University of Adelaide/International ; DAS00094//Grains Research and Development Corporation/International ; DAS00155//Grains Research and Development Corporation/International ; DAS00155//Grains Research and Development Corporation/International ; DAS00155//Grains Research and Development Corporation/International ; DP120100047//Australian Research Council/International ; FT140101303//Australian Research Council/International ; }, mesh = {Animals ; Australia ; Biological Assay ; Crosses, Genetic ; DNA, Mitochondrial/genetics ; Female ; Fertility ; *Genetic Variation ; Genetics, Population ; Geography ; Haplotypes/genetics ; Heterozygote ; *Hybridization, Genetic/drug effects ; Insecticide Resistance/drug effects/genetics ; Insecticides/toxicity ; Likelihood Functions ; Male ; Mitochondria/genetics ; Moths/*genetics/microbiology ; Phylogeny ; Species Specificity ; Sympatry ; Wolbachia/drug effects/physiology ; }, abstract = {BACKGROUND: Understanding genomic and phenotypic diversity among cryptic pest taxa has important implications for the management of pests and diseases. The diamondback moth, Plutella xylostella L., has been intensively studied due to its ability to evolve insecticide resistance and status as the world's most destructive pest of brassicaceous crops. The surprise discovery of a cryptic species endemic to Australia, Plutella australiana Landry & Hebert, raised questions regarding the distribution, ecological traits and pest status of the two species, the capacity for gene flow and whether specific management was required. Here, we collected Plutella from wild and cultivated brassicaceous plants from 75 locations throughout Australia and screened 1447 individuals to identify mtDNA lineages and Wolbachia infections. We genotyped genome-wide SNP markers using RADseq in coexisting populations of each species. In addition, we assessed reproductive compatibility in crossing experiments and insecticide susceptibility phenotypes using bioassays.

RESULTS: The two Plutella species coexisted on wild brassicas and canola crops, but only 10% of Plutella individuals were P. australiana. This species was not found on commercial Brassica vegetable crops, which are routinely sprayed with insecticides. Bioassays found that P. australiana was 19-306 fold more susceptible to four commonly-used insecticides than P. xylostella. Laboratory crosses revealed that reproductive isolation was incomplete but directionally asymmetric between the species. However, genome-wide nuclear SNPs revealed striking differences in genetic diversity and strong population structure between coexisting wild populations of each species. Nuclear diversity was 1.5-fold higher in P. australiana, yet both species showed limited variation in mtDNA. Infection with a single Wolbachia subgroup B strain was fixed in P. australiana, suggesting that a selective sweep contributed to low mtDNA diversity, while a subgroup A strain infected just 1.5% of P. xylostella.

CONCLUSIONS: Despite sympatric distributions and the capacity to hybridize, strong genomic and phenotypic divergence exists between these Plutella species that is consistent with contrasting colonization histories and reproductive isolation after secondary contact. Although P. australiana is a potential pest of brassicaceous crops, it is of secondary importance to P. xylostella.}, } @article {pmid29807401, year = {2018}, author = {Ali, H and Muhammad, A and Hou, Y}, title = {Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping.}, journal = {Journal of microbiology and biotechnology}, volume = {28}, number = {5}, pages = {796-808}, doi = {10.4014/jmb.1712.12019}, pmid = {29807401}, issn = {1738-8872}, mesh = {Animals ; Bacterial Load/*genetics ; Coleoptera/*microbiology ; DNA, Bacterial/analysis/genetics ; Female ; Genotype ; Life Cycle Stages ; Male ; Multilocus Sequence Typing ; Real-Time Polymerase Chain Reaction ; *Rickettsiaceae Infections/microbiology/veterinary ; Symbiosis/*genetics ; *Wolbachia/genetics/physiology ; }, abstract = {The intracellular bacterium Wolbachia pipientis is widespread in arthropods. Recently, possibilities of novel Wolbachia-mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting Wolbachia surface protein (wsp) and multilocus sequence type (MLST) genotypic systems, we determined the Wolbachia infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of B. longissima were infected with the same Wolbachia strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles (ftsZ-234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of B. longissima and Wolbachia. The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the wsp and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the Wolbachia endosymbiont in B. longissima, which demonstrated that Wolbachia is ubiquitous across all developmental stages and distributed in the entire body of B. longissima. Understanding the Wolbachia infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.}, } @article {pmid29801436, year = {2018}, author = {Opatovsky, I and Santos-Garcia, D and Ruan, Z and Lahav, T and Ofaim, S and Mouton, L and Barbe, V and Jiang, J and Zchori-Fein, E and Freilich, S}, title = {Modeling trophic dependencies and exchanges among insects' bacterial symbionts in a host-simulated environment.}, journal = {BMC genomics}, volume = {19}, number = {1}, pages = {402}, pmid = {29801436}, issn = {1471-2164}, support = {1481/13//Israel Science Foundation/ ; 484/17//Israel Science Foundation/ ; }, mesh = {Animals ; Bacteria/*genetics/*metabolism ; *Environment ; Genome, Bacterial/genetics ; Hemiptera/*microbiology ; Metabolic Networks and Pathways ; *Models, Biological ; *Symbiosis ; }, abstract = {BACKGROUND: Individual organisms are linked to their communities and ecosystems via metabolic activities. Metabolic exchanges and co-dependencies have long been suggested to have a pivotal role in determining community structure. In phloem-feeding insects such metabolic interactions with bacteria enable complementation of their deprived nutrition. The phloem-feeding whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) harbors an obligatory symbiotic bacterium, as well as varying combinations of facultative symbionts. This well-defined bacterial community in B. tabaci serves here as a case study for a comprehensive and systematic survey of metabolic interactions within the bacterial community and their associations with documented occurrences of bacterial combinations. We first reconstructed the metabolic networks of five common B. tabaci symbionts genera (Portiera, Rickettsia, Hamiltonella, Cardinium and Wolbachia), and then used network analysis approaches to predict: (1) species-specific metabolic capacities in a simulated bacteriocyte-like environment; (2) metabolic capacities of the corresponding species' combinations, and (3) dependencies of each species on different media components.

RESULTS: The predictions for metabolic capacities of the symbionts in the host environment were in general agreement with previously reported genome analyses, each focused on the single-species level. The analysis suggests several previously un-reported routes for complementary interactions and estimated the dependency of each symbiont in specific host metabolites. No clear association was detected between metabolic co-dependencies and co-occurrence patterns.

CONCLUSIONS: The analysis generated predictions for testable hypotheses of metabolic exchanges and co-dependencies in bacterial communities and by crossing them with co-occurrence profiles, contextualized interaction patterns into a wider ecological perspective.}, } @article {pmid29794009, year = {2018}, author = {Bakovic, V and Schebeck, M and Telschow, A and Stauffer, C and Schuler, H}, title = {Spatial spread of Wolbachia in Rhagoletis cerasi populations.}, journal = {Biology letters}, volume = {14}, number = {5}, pages = {}, pmid = {29794009}, issn = {1744-957X}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Czech Republic ; Hungary ; *Spatial Analysis ; Tephritidae/*microbiology ; Wolbachia/*physiology ; }, abstract = {The bacterial endosymbiont Wolbachia has been used to control insect pests owing to its ability to manipulate their life history and suppress infectious diseases. Therefore, knowledge on Wolbachia dynamics in natural populations is fundamental. The European cherry fruit fly, Rhagoletis cerasi, is infected with the Wolbachia strain wCer2, mainly present in southern and central European populations, and is currently spreading into wCer2-uninfected populations driven by high unidirectional cytoplasmic incompatibility. Here, we describe the distribution of wCer2 along two transition zones where the infection is spreading into wCer2-uninfected R. cerasi populations. Fine-scale sampling of 19 populations in the Czech Republic showed a smooth decrease of wCer2 frequency from south to north within a distance of less than 20 km. Sampling of 12 Hungarian populations, however, showed a sharp decline of wCer2 infection frequency within a few kilometres. We fitted a standard wave equation to our empirical data and estimated a Wolbachia wave speed of 1.9 km yr[-1] in the Czech Republic and 1.0 km yr[-1] in Hungary. Considering the univoltine life cycle and limited dispersal ability of R. cerasi, our study highlights a rapid Wolbachia spread in natural host populations.}, } @article {pmid29792944, year = {2018}, author = {Stouthamer, CM and Kelly, S and Hunter, MS}, title = {Enrichment of low-density symbiont DNA from minute insects.}, journal = {Journal of microbiological methods}, volume = {151}, number = {}, pages = {16-19}, doi = {10.1016/j.mimet.2018.05.013}, pmid = {29792944}, issn = {1872-8359}, mesh = {Animals ; Arthropods/genetics/microbiology ; Bacteria/*genetics ; DNA, Bacterial/*isolation & purification ; Host Microbial Interactions/*genetics ; Insecta/genetics/*microbiology ; Molecular Biology/*methods ; Sequence Analysis, DNA ; *Symbiosis ; Wasps/genetics/microbiology ; Wolbachia/genetics ; }, abstract = {Symbioses between bacteria and insects are often associated with changes in important biological traits that can significantly affect host fitness. To a large extent, studies of these interactions have been based on physiological changes or induced phenotypes in the host, and the genetic mechanisms by which symbionts interact with their hosts have only recently become better understood. Learning about symbionts has been challenging in part due to difficulties such as obtaining enough high quality genomic material for high throughput sequencing technology, especially for symbionts present in low titers, and in small or difficult to rear non-model hosts. Here we introduce a new method that substantially increases the yield of bacterial DNA in minute arthropod hosts, and requires less starting material relative to previous published methods.}, } @article {pmid29789369, year = {2018}, author = {Schultz, MJ and Tan, AL and Gray, CN and Isern, S and Michael, SF and Frydman, HM and Connor, JH}, title = {Wolbachia wStri Blocks Zika Virus Growth at Two Independent Stages of Viral Replication.}, journal = {mBio}, volume = {9}, number = {3}, pages = {}, pmid = {29789369}, issn = {2150-7511}, support = {R01 AI099210/AI/NIAID NIH HHS/United States ; R21 NS101151/NS/NINDS NIH HHS/United States ; R56 AI097589/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*virology ; Animals ; *Antibiosis ; Chikungunya virus/genetics/growth & development/physiology ; Dengue Virus/genetics/growth & development/physiology ; Male ; Mosquito Vectors/*virology ; Virus Internalization ; *Virus Replication ; Wolbachia/genetics/*physiology ; Zika Virus/genetics/growth & development/*physiology ; }, abstract = {Mosquito-transmitted viruses are spread globally and present a great risk to human health. Among the many approaches investigated to limit the diseases caused by these viruses are attempts to make mosquitos resistant to virus infection. Coinfection of mosquitos with the bacterium Wolbachia pipientis from supergroup A is a recent strategy employed to reduce the capacity for major vectors in the Aedes mosquito genus to transmit viruses, including dengue virus (DENV), Chikungunya virus (CHIKV), and Zika virus (ZIKV). Recently, a supergroup B Wolbachia wStri, isolated from Laodelphax striatellus, was shown to inhibit multiple lineages of ZIKV in Aedes albopictus cells. Here, we show that wStri blocks the growth of positive-sense RNA viruses DENV, CHIKV, ZIKV, and yellow fever virus by greater than 99.9%. wStri presence did not affect the growth of the negative-sense RNA viruses LaCrosse virus or vesicular stomatitis virus. Investigation of the stages of the ZIKV life cycle inhibited by wStri identified two distinct blocks in viral replication. We found a reduction of ZIKV entry into wStri-infected cells. This was partially rescued by the addition of a cholesterol-lipid supplement. Independent of entry, transfected viral genome was unable to replicate in Wolbachia-infected cells. RNA transfection and metabolic labeling studies suggested that this replication defect is at the level of RNA translation, where we saw a 66% reduction in mosquito protein synthesis in wStri-infected cells. This study's findings increase the potential for application of wStri to block additional arboviruses and also identify specific blocks in viral infection caused by Wolbachia coinfection.IMPORTANCE Dengue, Zika, and yellow fever viruses are mosquito-transmitted diseases that have spread throughout the world, causing millions of infections and thousands of deaths each year. Existing programs that seek to contain these diseases through elimination of the mosquito population have so far failed, making it crucial to explore new ways of limiting the spread of these viruses. Here, we show that introduction of an insect symbiont, Wolbachia wStri, into mosquito cells is highly effective at reducing yellow fever virus, dengue virus, Zika virus, and Chikungunya virus production. Reduction of virus replication was attributable to decreases in entry and a strong block of virus gene expression at the translational level. These findings expand the potential use of Wolbachia wStri to block viruses and identify two separate steps for limiting virus replication in mosquitos that could be targeted via microbes or other means as an antiviral strategy.}, } @article {pmid29779872, year = {2018}, author = {Funkhouser-Jones, LJ and van Opstal, EJ and Sharma, A and Bordenstein, SR}, title = {The Maternal Effect Gene Wds Controls Wolbachia Titer in Nasonia.}, journal = {Current biology : CB}, volume = {28}, number = {11}, pages = {1692-1702.e6}, pmid = {29779872}, issn = {1879-0445}, support = {R21 HD086833/HD/NICHD NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; S10 OD021630/OD/NIH HHS/United States ; P30 EY008126/EY/NEI NIH HHS/United States ; P30 DK020593/DK/NIDDK NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; U24 DK059637/DK/NIDDK NIH HHS/United States ; P30 CA068485/CA/NCI NIH HHS/United States ; T32 GM008554/GM/NIGMS NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Biological Evolution ; Insect Proteins/*genetics/metabolism ; Maternal Inheritance ; Quantitative Trait Loci/genetics ; Selection, Genetic ; Sequence Alignment ; Species Specificity ; Symbiosis/*genetics ; Wasps/*genetics/*microbiology ; Wolbachia/*physiology ; }, abstract = {Maternal transmission of intracellular microbes is pivotal in establishing long-term, intimate symbioses. For germline microbes that exert negative reproductive effects on their hosts, selection can theoretically favor the spread of host genes that counteract the microbe's harmful effects. Here, we leverage a major difference in bacterial (Wolbachia pipientis) titers between closely related wasp species with forward genetic, transcriptomic, and cytological approaches to map two quantitative trait loci that suppress bacterial titers via a maternal effect. Fine mapping and knockdown experiments identify the gene Wolbachia density suppressor (Wds), which dominantly suppresses bacterial transmission from mother to embryo. Wds evolved by lineage-specific non-synonymous changes driven by positive selection. Collectively, our findings demonstrate that a genetically simple change arose by positive Darwinian selection in less than a million years to regulate maternally transmitted bacteria via a dominant, maternal effect gene.}, } @article {pmid29777177, year = {2018}, author = {Flores, HA and O'Neill, SL}, title = {Controlling vector-borne diseases by releasing modified mosquitoes.}, journal = {Nature reviews. Microbiology}, volume = {16}, number = {8}, pages = {508-518}, pmid = {29777177}, issn = {1740-1534}, support = {102591/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*genetics/*microbiology ; Animals ; Genetic Engineering ; *Mosquito Control ; Mosquito Vectors/genetics/microbiology ; Virus Diseases/prevention & control ; }, abstract = {Aedes mosquito-transmitted diseases, such as dengue, Zika and chikungunya, are becoming major global health emergencies while old threats, such as yellow fever, are re-emerging. Traditional control methods, which have focused on reducing mosquito populations through the application of insecticides or preventing breeding through removal of larval habitat, are largely ineffective, as evidenced by the increasing global disease burden. Here, we review novel mosquito population reduction and population modification approaches with a focus on control methods based on the release of mosquitoes, including the release of Wolbachia-infected mosquitoes and strategies to genetically modify the vector, that are currently under development and have the potential to contribute to a reversal of the current alarming disease trends.}, } @article {pmid29776407, year = {2018}, author = {Lindsey, ARI and Kelkar, YD and Wu, X and Sun, D and Martinson, EO and Yan, Z and Rugman-Jones, PF and Hughes, DST and Murali, SC and Qu, J and Dugan, S and Lee, SL and Chao, H and Dinh, H and Han, Y and Doddapaneni, HV and Worley, KC and Muzny, DM and Ye, G and Gibbs, RA and Richards, S and Yi, SV and Stouthamer, R and Werren, JH}, title = {Comparative genomics of the miniature wasp and pest control agent Trichogramma pretiosum.}, journal = {BMC biology}, volume = {16}, number = {1}, pages = {54}, pmid = {29776407}, issn = {1741-7007}, support = {U54 HG003273/HG/NHGRI NIH HHS/United States ; }, mesh = {Animals ; *Evolution, Molecular ; Genomics ; Moths/parasitology ; *Pest Control, Biological ; Phylogeny ; Wasps/*classification/*genetics/pathogenicity ; Whole Genome Sequencing/methods ; }, abstract = {BACKGROUND: Trichogrammatids are minute parasitoid wasps that develop within other insect eggs. They are less than half a millimeter long, smaller than some protozoans. The Trichogrammatidae are one of the earliest branching families of Chalcidoidea: a diverse superfamily of approximately half a million species of parasitoid wasps, proposed to have evolved from a miniaturized ancestor. Trichogramma are frequently used in agriculture, released as biological control agents against major moth and butterfly pests. Additionally, Trichogramma are well known for their symbiotic bacteria that induce asexual reproduction in infected females. Knowledge of the genome sequence of Trichogramma is a major step towards further understanding its biology and potential applications in pest control.

RESULTS: We report the 195-Mb genome sequence of Trichogramma pretiosum and uncover signatures of miniaturization and adaptation in Trichogramma and related parasitoids. Comparative analyses reveal relatively rapid evolution of proteins involved in ribosome biogenesis and function, transcriptional regulation, and ploidy regulation. Chalcids also show loss or especially rapid evolution of 285 gene clusters conserved in other Hymenoptera, including many that are involved in signal transduction and embryonic development. Comparisons between sexual and asexual lineages of Trichogramma pretiosum reveal that there is no strong evidence for genome degradation (e.g., gene loss) in the asexual lineage, although it does contain a lower repeat content than the sexual lineage. Trichogramma shows particularly rapid genome evolution compared to other hymenopterans. We speculate these changes reflect adaptations to miniaturization, and to life as a specialized egg parasitoid.

CONCLUSIONS: The genomes of Trichogramma and related parasitoids are a valuable resource for future studies of these diverse and economically important insects, including explorations of parasitoid biology, symbiosis, asexuality, biological control, and the evolution of miniaturization. Understanding the molecular determinants of parasitism can also inform mass rearing of Trichogramma and other parasitoids for biological control.}, } @article {pmid29771340, year = {2018}, author = {Schebeck, M and Feldkirchner, L and Marín, B and Krumböck, S and Schuler, H and Stauffer, C}, title = {Reproductive Manipulators in the Bark Beetle Pityogenes chalcographus (Coleoptera: Curculionidae)-The Role of Cardinium, Rickettsia, Spiroplasma, and Wolbachia.}, journal = {Journal of insect science (Online)}, volume = {18}, number = {3}, pages = {}, pmid = {29771340}, issn = {1536-2442}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Female ; Male ; Polymerase Chain Reaction ; Reproduction ; Rickettsia/*isolation & purification ; Spiroplasma/*isolation & purification ; *Symbiosis ; Weevils/*microbiology ; Wolbachia/*isolation & purification ; }, abstract = {Heritable bacterial endosymbionts can alter the biology of numerous arthropods. They can influence the reproductive outcome of infected hosts, thus affecting the ecology and evolution of various arthropod species. The spruce bark beetle Pityogenes chalcographus (L.) (Coleoptera: Curculionidae: Scolytinae) was reported to express partial, unidirectional crossing incompatibilities among certain European populations. Knowledge on the background of these findings is lacking; however, bacterial endosymbionts have been assumed to manipulate the reproduction of this beetle. Previous work reported low-density and low-frequency Wolbachia infections of P. chalcographus but found it unlikely that this infection results in reproductive alterations. The aim of this study was to test the hypothesis of an endosymbiont-driven incompatibility, other than Wolbachia, reflected by an infection pattern on a wide geographic scale. We performed a polymerase chain reaction (PCR) screening of 226 individuals from 18 European populations for the presence of the endosymbionts Cardinium, Rickettsia, and Spiroplasma, and additionally screened these individuals for Wolbachia. Positive PCR products were sequenced to characterize these bacteria. Our study shows a low prevalence of these four endosymbionts in P. chalcographus. We detected a yet undescribed Spiroplasma strain in a single individual from Greece. This is the first time that this endosymbiont has been found in a bark beetle. Further, Wolbachia was detected in three beetles from two Scandinavian populations and two new Wolbachia strains were described. None of the individuals analyzed were infected with Cardinium and Rickettsia. The low prevalence of bacteria found here does not support the hypothesis of an endosymbiont-driven reproductive incompatibility in P. chalcographus.}, } @article {pmid29766491, year = {2018}, author = {Kriesner, P and Hoffmann, AA}, title = {Rapid spread of a Wolbachia infection that does not affect host reproduction in Drosophila simulans cage populations.}, journal = {Evolution; international journal of organic evolution}, volume = {}, number = {}, pages = {}, doi = {10.1111/evo.13506}, pmid = {29766491}, issn = {1558-5646}, abstract = {Wolbachia endosymbionts that are maternally inherited can spread rapidly in host populations through inducing sterility in uninfected females, but some Wolbachia infections do not influence host reproduction yet still persist. These infections are particularly interesting because they likely represent mutualistic endosymbionts, spreading by increasing host fitness. Here, we document such a spread in the wAu infection of Drosophila simulans. By establishing multiple replicate cage populations, we show that wAu consistently increased from an intermediate frequency to near fixation, representing an estimated fitness advantage of around 20% for infected females. The effective population size in the cages was estimated from SNP markers to be around a few thousand individuals, precluding large effects of genetic drift in the populations. The exact reasons for the fitness advantage are unclear but viral protection and nutritional benefits are two possibilities.}, } @article {pmid29765368, year = {2018}, author = {Hu, W and Kuang, F and Lu, Z and Zhang, N and Chen, T}, title = {Killing Effects of an Isolated Serratia marcescens KH-001 on Diaphorina citri via Lowering the Endosymbiont Numbers.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {860}, pmid = {29765368}, issn = {1664-302X}, abstract = {Huanglongbing (HLB) is the most devastating citrus disease worldwide, and suppression of the Asian citrus psyllid (Diaphorina citri) is regarded as an effective method to inhibit the spread of HLB. In this study, we isolated a strain named as Serratia marcescens KH-001 from D. citri nymphs suffering from disease, and evaluated its killing effect on D. citri via toxicity test and effect on microbial community in D. citri using high-throughput sequencing. Our results indicated that S. marcescens KH-001 could effectively kill 83% of D. citri nymphs, while the fermentation products of S. marcescens KH-001 only killed 40% of the D. citrinymphs. High-throughput sequencing results indicated that the S. marcescens KH-001 increased the OTU numbers from 62.5 (PBS buffer) to 81.5, while significantly lowered the Shannon index compared with Escherichia coli DH5α (group E) (p < 0.05). OTU analysis showed that the S. marcescens KH-001 had significantly reduced the relative abundance of endosymbionts Wolbachia, Profftella, and Carsonella in group S compared with that in other groups (p < 0.05). Therefore, the direct killing effect of the fermentation products of S. marcescens KH-001 and the indirect effect via reducing the numbers of endosymbionts (Wolbachia, Profftella, and Carsonella) of D. citri endow S. marcescens KH-001 a sound killing effect on D. citri. Further work need to do before this strain is used as a sound biological control agents.}, } @article {pmid29761037, year = {2018}, author = {Duplouy, A and Hornett, EA}, title = {Uncovering the hidden players in Lepidoptera biology: the heritable microbial endosymbionts.}, journal = {PeerJ}, volume = {6}, number = {}, pages = {e4629}, pmid = {29761037}, issn = {2167-8359}, abstract = {The Lepidoptera is one of the most widespread and recognisable insect orders. Due to their remarkable diversity, economic and ecological importance, moths and butterflies have been studied extensively over the last 200 years. More recently, the relationship between Lepidoptera and their heritable microbial endosymbionts has received increasing attention. Heritable endosymbionts reside within the host's body and are often, but not exclusively, inherited through the female line. Advancements in molecular genetics have revealed that host-associated microbes are both extremely prevalent among arthropods and highly diverse. Furthermore, heritable endosymbionts have been repeatedly demonstrated to play an integral role in many aspects of host biology, particularly host reproduction. Here, we review the major findings of research of heritable microbial endosymbionts of butterflies and moths. We promote the Lepidoptera as important models in the study of reproductive manipulations employed by heritable endosymbionts, with the mechanisms underlying male-killing and feminisation currently being elucidated in moths and butterflies. We also reveal that the vast majority of research undertaken of Lepidopteran endosymbionts concerns Wolbachia. While this highly prevalent bacterium is undoubtedly important, studies should move towards investigating the presence of other, and interacting endosymbionts, and we discuss the merits of examining the microbiome of Lepidoptera to this end. We finally consider the importance of understanding the influence of endosymbionts under global environmental change and when planning conservation management of endangered Lepidoptera species.}, } @article {pmid29760316, year = {2018}, author = {Katsuma, S and Kiuchi, T and Kawamoto, M and Fujimoto, T and Sahara, K}, title = {Unique sex determination system in the silkworm, Bombyx mori: current status and beyond.}, journal = {Proceedings of the Japan Academy. Series B, Physical and biological sciences}, volume = {94}, number = {5}, pages = {205-216}, pmid = {29760316}, issn = {1349-2896}, mesh = {Animals ; Base Sequence ; Bombyx/*genetics/microbiology/physiology ; Chromosomes, Insect/genetics ; Gene Expression Profiling ; Sex Characteristics ; *Sex Determination Processes ; Wolbachia/physiology ; }, abstract = {The silkworm Bombyx mori has been used for silk production for over 5,000 years. In addition to its contribution to sericulture, B. mori has played an important role in the field of genetics. Classical genetic studies revealed that a gene(s) with a strong feminizing activity is located on the W chromosome, but this W-linked feminizing gene, called Feminizer (Fem), had not been cloned despite more than 80 years of study. In 2014, we discovered that Fem is a precursor of a single W chromosome-derived PIWI-interacting RNA (piRNA). Fem-derived piRNA binds to PIWI protein, and this complex then cleaves the mRNA of the Z-linked Masculinizer (Masc) gene, which encodes a protein required for both masculinization and dosage compensation. These findings showed that the piRNA-mediated interaction between the two sex chromosomes is the primary signal for the sex determination cascade in B. mori. In this review, we summarize the history, current status, and perspective of studies on sex determination and related topics in B. mori.}, } @article {pmid29757522, year = {2018}, author = {Morçiçek, B and Taskin, BG and Doğaç, E and Doğaroğlu, T and Taskin, V}, title = {Evidence of natural Wolbachia infections and molecular identification of field populations of Culex pipiens complex (Diptera: Culicidae) mosquitoes in western Turkey.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {43}, number = {1}, pages = {44-51}, doi = {10.1111/jvec.12281}, pmid = {29757522}, issn = {1948-7134}, mesh = {Animals ; Culex/*microbiology ; Cytochromes c/metabolism ; Cytoplasm/metabolism ; Polymerase Chain Reaction ; Turkey ; Wolbachia/*pathogenicity ; }, abstract = {Establishing reliable risk projection information about the distribution pattern of members of the Culex pipiens complex is of particular interest, as these mosquitoes are competent vectors for certain disease-causing pathogens. Wolbachia, a maternally inherited bacterial symbiont, are distributed in various arthropod species and can induce cytoplasmic incompatibility, i.e., reduced egg hatch, in certain crosses. It is being considered as a tool for population control of mosquito disease vectors. The Aegean region is characterized by highly populated, rural, and agricultural areas and is also on the route of the migratory birds. In this study, a fragment of the 658 bp of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene, which includes the barcode region, was employed to differentiate Cx. pipiens complex species found in this region. Moreover, for the first time, the prevalence of Wolbachia endobacteria in these natural populations was examined using PCR amplification of a specific wsp gene. Our results revealed a widespread (more than 90%, n=121) presence of the highly efficient West Nile virus vector Cx. quinquefasciatus in the region. We also found that Wolbachia infection is widespread; the average prevalence was 62% in populations throughout the region. This study provided valuable information about the composition of Cx. pipiens complex mosquitoes and the prevalence of Wolbachia infection in these populations in the Aegean region. This information will be helpful in tracking mosquito-borne diseases and designing and implementing Wolbachia-based control strategies in the region.}, } @article {pmid29751814, year = {2018}, author = {Ant, TH and Sinkins, SP}, title = {A Wolbachia triple-strain infection generates self-incompatibility in Aedes albopictus and transmission instability in Aedes aegypti.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {295}, pmid = {29751814}, issn = {1756-3305}, support = {202888, 108508/WT_/Wellcome Trust/United Kingdom ; MC_PC_15087/MRC_/Medical Research Council/United Kingdom ; ZK/16-021/MRC_/Medical Research Council/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; BB/K004506/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Aedes/genetics/*microbiology/physiology ; Animals ; Female ; Insect Vectors/genetics/microbiology/physiology ; Male ; Reproduction ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Artificially-introduced transinfections of the intracellular bacterium Wolbachia pipientis have the potential to reduce the vectorial capacity of mosquito populations for viruses such as dengue and chikungunya. Aedes albopictus has two native strains of Wolbachia, but their replacement with the non-native wMel strain blocks transmission of both viruses. The pattern of cytoplasmic incompatiiblity generated by wMel with wild-types is bidirectional. Novel-plus-native-strain co-infection is predicted to lead to a more efficient population spread capacity; from a bi-directional to a uni-directional cytoplasmic incompatibility (CI) model.

RESULTS: A novel-plus-native-strain triple-infection in Ae. albopictus (wAlbAwAlbBwMel) was generated. Although triple-infected females were fully reproductively viable with uninfected males, they displayed self-incompatibility. qPCR of specific strains in dissected tissues suggested that this may be due to the displacement of one of the native strains (wAlbA) from the ovaries of triple-infected females. When the triple strain infection was transferred into Aedes aegypti it displayed an unexpectedly low level of transmission fidelity of the three strains in this species.

CONCLUSIONS: These results suggest that combining Wolbachia strains can lead to co-infection interactions that can affect outcomes of CI and maternal transmission.}, } @article {pmid29745300, year = {2018}, author = {Karimian, F and Vatandoost, H and Rassi, Y and Maleki-Ravasan, N and Choubdar, N and Koosha, M and Arzamani, K and Moradi-Asl, E and Veysi, A and Alipour, H and Shirani, M and Oshaghi, MA}, title = {wsp-based analysis of Wolbachia strains associated with Phlebotomus papatasi and P. sergenti (Diptera: Psychodidae) main cutaneous leishmaniasis vectors, introduction of a new subgroup wSerg.}, journal = {Pathogens and global health}, volume = {112}, number = {3}, pages = {152-160}, pmid = {29745300}, issn = {2047-7732}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Cluster Analysis ; Genotype ; Iran ; Phlebotomus/*microbiology ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; *Serogroup ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Sand flies of Phlebotomus papatasi and P. sergenti are the main vectors of cutaneous leishmanisis (CL) in the old world. We aimed to screen Iranian P. papatasi and P. sergenti for their natural infections with Wolbachia and to determine their phylogenetic association with other species. Wolbachia surface protein (wsp) gene was PCR amplified from DNA extracted from Phlebotomus species, sequenced, and were analysed in combination with wsp sequences related to Phelebtominae and other insects. All Wolbachia-infecting Iranian sand flies of P. papatasi and P. sergenti were classified in the Supergroup A., Wolbachia isolated from P. sergenti were clustered in a new subgroup within Supergroup A so-called wSreg. The Wolbachia strains identified from the P. papatasi clustered mainly in the subgroup wPap and partly in wSerg. Multiple Wholbachia infection within a single population of P.papatasi warrants investigation on existence and intensity of cytoplasmic incompatibility between the wPap and wSerg subgroups.}, } @article {pmid29732657, year = {2018}, author = {Zheng, Y and Bi, J and Hou, MY and Shen, W and Zhang, W and Ai, H and Yu, XQ and Wang, YF}, title = {Ocnus is essential for male germ cell development in Drosophila melanogaster.}, journal = {Insect molecular biology}, volume = {27}, number = {5}, pages = {545-555}, doi = {10.1111/imb.12393}, pmid = {29732657}, issn = {1365-2583}, mesh = {Animals ; Drosophila Proteins/*physiology ; Drosophila melanogaster/*physiology ; Fertility ; *Gene Expression Regulation ; Male ; Phosphoric Monoester Hydrolases/*physiology ; *Spermatogenesis ; Testis/growth & development ; Transcriptome ; }, abstract = {The ocnus (ocn) gene encodes a protein abundant in the testes, implying its role in testis development. When Drosophila melanogaster is infected with the endosymbiont wMel Wolbachia, which affects the spermatogenesis of its hosts, ocn is downregulated in the third-instar larval testes, suggesting a role of ocn in spermatogenesis. In this study, we knocked down ocn in the testes and found that the hatch rates of embryos derived from ocn-knockdown males were significantly decreased, and 84.38% of the testes were much smaller in comparison to controls. Analysis of the smaller testes showed no germ cells but they had an extended hub. Using RNA-sequencing (RNA-Seq), we identified 69 genes with at least a twofold change (q-value < 5%) in their expression after ocn knockdown; of these, eight testes-specific and three reproduction-related genes were verified to be significantly downregulated using quantitative reverse transcription-PCR. Three genes (orientation disruptor, p24-2 and CG13541) were also significantly downregulated in the presence of Wolbachia. Furthermore, 98 genes were not expressed when ocn was knocked down in testes. These results suggest that ocn plays a crucial role in male germ cell development in Drosophila, possibly by regulating the expression of multiple spermatogenesis-related genes. Our data provide important information to help understand the molecular regulatory mechanisms underlying spermatogenesis.}, } @article {pmid29725059, year = {2018}, author = {Dittmer, J and Bouchon, D}, title = {Feminizing Wolbachia influence microbiota composition in the terrestrial isopod Armadillidium vulgare.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {6998}, pmid = {29725059}, issn = {2045-2322}, mesh = {Animal Structures/*microbiology ; Animals ; Bacteria/classification/genetics ; Isopoda/*microbiology ; Metagenomics ; Microbial Interactions ; *Microbiota ; Wolbachia/*growth & development ; }, abstract = {Wolbachia are widespread heritable endosymbionts of arthropods notorious for their profound effects on host fitness as well as for providing protection against viruses and eukaryotic parasites, indicating that they can interact with other microorganisms sharing the same host environment. Using the terrestrial isopod crustacean Armadillidium vulgare, its highly diverse microbiota (>200 bacterial genera) and its three feminizing Wolbachia strains (wVulC, wVulM, wVulP) as a model system, the present study demonstrates that Wolbachia can even influence the composition of a diverse bacterial community under both laboratory and natural conditions. While host origin is the major determinant of the taxonomic composition of the microbiota in A. vulgare, Wolbachia infection affected both the presence and, more importantly, the abundance of many bacterial taxa within each host population, possibly due to competitive interactions. Moreover, different Wolbachia strains had different impacts on microbiota composition. As such, infection with wVulC affected a higher number of taxa than infection with wVulM, possibly due to intrinsic differences in virulence and titer between these two strains. In conclusion, this study shows that heritable endosymbionts such as Wolbachia can act as biotic factors shaping the microbiota of arthropods, with as yet unknown consequences on host fitness.}, } @article {pmid29722798, year = {2018}, author = {Li, H and Li, T and Qu, J}, title = {Stochastic processes govern bacterial communities from the blood of pikas and from their arthropod vectors.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {6}, pages = {}, doi = {10.1093/femsec/fiy082}, pmid = {29722798}, issn = {1574-6941}, mesh = {Animals ; Arthropod Vectors/*microbiology ; Bacteria/*classification/genetics/*isolation & purification ; Bacterial Infections/microbiology/*transmission ; Gastrointestinal Microbiome/*physiology ; High-Throughput Nucleotide Sequencing ; Humans ; Lagomorpha/blood/*microbiology ; Microbiota/genetics ; RNA, Ribosomal, 16S/genetics ; Siphonaptera/*microbiology ; Stochastic Processes ; }, abstract = {Vector-borne microbes influence pathogen transmission and blood microbiomes, thereby affecting the emergence of infectious diseases. Thus, understanding the relationship between host and vector microbiomes is of importance. In this study, we investigated the bacterial community composition, diversity and assembly of the flea (Rhadinopsylla dahurica vicina), torsalo (Hypoderma curzonial), and the blood and gut of their shared pika host, Ochotona curzoniae. Bartonella, Sphingomonas and Bradyrhizobium were enriched in blood, while Wolbachia and Fusobacterium were more abundant in fleas and torsaloes. Most of potential pathogenic microbes (belonging to Fusobacterium, Rickettsia, Kingella, Porphyromonas, Bartonella and Mycoplasma) were present in the blood of pikas and their vectors. Blood communities were more similar to those from fleas than other sample types and were independent of host factors or geographical sites. Notably, blood microbes originate mainly from fleas rather than gut or torsaloes. Interestingly, the community assembly of blood, fleas or torsaloes was primarily governed by stochastic processes, while the gut microbiome was determined by deterministic processes. Ecological drift plays a dominant role in the assembly of blood and flea microbiomes. These results reflect the difficulty for predicting and regulating the microbial ecology of fleas for the prevention of potential microbiome-associated diseases.}, } @article {pmid29720714, year = {2018}, author = {Pereira, TN and Rocha, MN and Sucupira, PHF and Carvalho, FD and Moreira, LA}, title = {Wolbachia significantly impacts the vector competence of Aedes aegypti for Mayaro virus.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {6889}, pmid = {29720714}, issn = {2045-2322}, mesh = {Aedes/microbiology/*virology ; Animals ; Cell Line ; Cells, Cultured ; Female ; Humans ; Mosquito Vectors/microbiology/*virology ; Symbiosis ; Togaviridae/pathogenicity/*physiology ; Togaviridae Infections/transmission ; *Virus Replication ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia, an intracellular endosymbiont present in up to 70% of all insect species, has been suggested as a sustainable strategy for the control of arboviruses such as Dengue, Zika and Chikungunya. As Mayaro virus outbreaks have also been reported in Latin American countries, the objective of this study was to evaluate the vector competence of Brazilian field-collected Ae. aegypti and the impact of Wolbachia (wMel strain) upon this virus. Our in vitro studies with Aag2 cells showed that Mayaro virus can rapidly multiply, whereas in wMel-infected Aag2 cells, viral growth was significantly impaired. In addition, C6/36 cells seem to have alterations when infected by Mayaro virus. In vivo experiments showed that field-collected Ae. aegypti mosquitoes are highly permissive to Mayaro virus infection, and high viral prevalence was observed in the saliva. On the other hand, Wolbachia-harboring mosquitoes showed significantly impaired capability to transmit Mayaro virus. Our results suggest that the use of Wolbachia-harboring mosquitoes may represent an effective mechanism for the reduction of Mayaro virus transmission throughout Latin America.}, } @article {pmid29720706, year = {2018}, author = {Pance, A}, title = {Can Wolbachia save the day?.}, journal = {Nature reviews. Microbiology}, volume = {16}, number = {7}, pages = {396}, doi = {10.1038/s41579-018-0021-4}, pmid = {29720706}, issn = {1740-1534}, mesh = {Animals ; Culicidae/*microbiology/*parasitology ; Host-Pathogen Interactions ; Humans ; Malaria, Falciparum/*prevention & control/transmission ; Mosquito Control/*methods ; Plasmodium falciparum ; Wolbachia/*physiology ; }, } @article {pmid29709692, year = {2018}, author = {Steiner, FM and Csősz, S and Markó, B and Gamisch, A and Rinnhofer, L and Folterbauer, C and Hammerle, S and Stauffer, C and Arthofer, W and Schlick-Steiner, BC}, title = {Turning one into five: Integrative taxonomy uncovers complex evolution of cryptic species in the harvester ant Messor "structor".}, journal = {Molecular phylogenetics and evolution}, volume = {127}, number = {}, pages = {387-404}, doi = {10.1016/j.ympev.2018.04.005}, pmid = {29709692}, issn = {1095-9513}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Amplified Fragment Length Polymorphism Analysis ; Animals ; Ants/anatomy & histology/*classification/genetics/microbiology ; *Biological Evolution ; DNA, Mitochondrial/genetics ; Discriminant Analysis ; Ecosystem ; Female ; Male ; Models, Theoretical ; Phylogeny ; Principal Component Analysis ; Species Specificity ; Terminology as Topic ; Wolbachia/physiology ; }, abstract = {Seed harvesting ants are ecosystem engineers that shape vegetation, nutrient cycles, and microclimate. Progress in ecological research is, however, slowed down by poor species delimitation. For example, it has not been resolved to date, how many species the European harvester ant Messor "structor" (Latreille, 1798) represents. Since its first description, splitting into additional taxa was often proposed but not accepted later on due to inconsistent support from morphology and ecology. Here, we took an iterative integrative-taxonomy approach - comparing multiple, independent data sets of the same sample - and used traditional morphometrics, Wolbachia symbionts, mitochondrial DNA, amplified fragment length polymorphism, and ecological niche modelling. Using the complementarity of the data sets applied, we resolved multiple, strong disagreements over the number of species, ranging from four to ten, and the allocation of individuals to species. We consider most plausible a five-species hypothesis and conclude the taxonomic odyssey by redescribing Messor structor, M. ibericus Santschi, 1925, and M. muticus (Nylander, 1849) stat.rev., and by describing two new species, M. ponticus sp.n. and M. mcarthuri sp.n. The evolutionary explanations invoked in resolving the various data conflicts include pronounced morphological crypsis, incomplete lineage-sorting or ongoing cospeciation of endosymbionts, and peripatric speciation - these ants' significance to evolutionary biology parallels that to ecology. The successful solution of this particular problem illustrates the usefulness of the integrative approach to other systematic problems of comparable complexity and the importance of understanding evolution to drawing correct conclusions on species' attributes, including their ecology and biogeography.}, } @article {pmid29701280, year = {2018}, author = {Ote, M and Yamamoto, D}, title = {Enhancing Nanos expression via the bacterial TomO protein is a conserved strategy used by the symbiont Wolbachia to fuel germ stem cell maintenance in infected Drosophila females.}, journal = {Archives of insect biochemistry and physiology}, volume = {98}, number = {3}, pages = {e21471}, doi = {10.1002/arch.21471}, pmid = {29701280}, issn = {1520-6327}, mesh = {Animals ; Animals, Genetically Modified ; Bacterial Proteins/*physiology ; Culex/genetics/microbiology ; Drosophila/*microbiology/physiology ; Drosophila Proteins/*metabolism ; Female ; Germ Cells/*physiology ; Male ; RNA-Binding Proteins/*metabolism ; Structure-Activity Relationship ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The toxic manipulator of oogenesis (TomO) protein has been identified in the wMel strain of Wolbachia that symbioses with the vinegar fly Drosophila melanogaster, as a protein that affects host reproduction. TomO protects germ stem cells (GSCs) from degeneration, which otherwise occurs in ovaries of host females that are mutant for the gene Sex-lethal (Sxl). We isolated the TomO homologs from wPip, a Wolbachia strain from the mosquito Culex quinquefasciatus. One of the homologs, TomOwPip 1, exerted the GSC rescue activity in fly Sxl mutants when lacking its hydrophobic stretches. The GSC-rescuing action of the TomOwPip 1 variant was ascribable to its abilities to associate with Nanos (nos) mRNA and to enhance Nos protein expression. The analysis of structure-activity relationships with TomO homologs and TomO deletion variants revealed distinct modules in the protein that are each dedicated to different functions, i.e., subcellular localization, nos mRNA binding or Nos expression enhancement. We propose that modular reshuffling is the basis for structural and functional diversification of TomO protein members.}, } @article {pmid29694414, year = {2018}, author = {Bridgeman, B and Morgan-Richards, M and Wheeler, D and Trewick, SA}, title = {First detection of Wolbachia in the New Zealand biota.}, journal = {PloS one}, volume = {13}, number = {4}, pages = {e0195517}, pmid = {29694414}, issn = {1932-6203}, mesh = {Animals ; Bacterial Proteins/genetics ; Bayes Theorem ; Biological Evolution ; *Biota/genetics ; Computational Biology ; DNA, Bacterial/analysis ; High-Throughput Nucleotide Sequencing ; New Zealand ; Orthoptera/microbiology ; Phylogeny ; Wasps/microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia is one of the most widespread intracellular bacteria on earth, estimated to infect between 40 and 66% of arthropod species in most ecosystems that have been surveyed. Their significance rests not only in their vast distribution, but also in their ability to modify the reproductive biology of their hosts, which can ultimately affect genetic diversity and speciation of infected populations. Wolbachia has yet to be formally identified in the fauna of New Zealand which has high levels of endemic biodiversity and this represents a gap in our understanding of the global biology of Wolbachia. Using High Throughput Sequencing (HTS) of host DNA in conjunction with traditional molecular techniques we identified six endemic Orthoptera species that were positive for Wolbachia infection. In addition, short-sequence amplification with Wolbachia specific primers applied to New Zealand and introduced invertebrates detected a further 153 individuals positive for Wolbachia. From these short-range DNA amplification products sequence data was obtained for the ftsZ gene region from 86 individuals representing 10 host species. Phylogenetic analysis using the sequences obtained in this study reveals that there are two distinct Wolbachia bacteria lineages in New Zealand hosts belonging to recognised Wolbachia supergroups (A and B). These represent the first described instances of Wolbachia in the New Zealand native fauna, including detection in putative parasitoids of infected Orthoptera suggesting a possible transmission path. Our detection of Wolbachia infections of New Zealand species provides the opportunity to study local transmission of Wolbachia and explore their role in the evolution of New Zealand invertebrates.}, } @article {pmid29691935, year = {2018}, author = {Henry, LP and Newton, ILG}, title = {Mitochondria and Wolbachia titers are positively correlated during maternal transmission.}, journal = {Molecular ecology}, volume = {27}, number = {11}, pages = {2634-2646}, doi = {10.1111/mec.14700}, pmid = {29691935}, issn = {1365-294X}, mesh = {Animals ; Biological Evolution ; Drosophila melanogaster/genetics ; Female ; Gene Expression/genetics ; Genotype ; Infectious Disease Transmission, Vertical ; Mitochondria/*genetics ; NADH Dehydrogenase/genetics ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Mothers provide their offspring with symbionts. Maternally transmitted, intracellular symbionts must disperse from mother to offspring with other cytoplasmic elements, like mitochondria. Here, we investigated how the intracellular symbiont Wolbachia interacts with mitochondria during maternal transmission. Mitochondria and Wolbachia may interact antagonistically and compete as each population tries to ensure its own evolutionary success. Alternatively, mitochondria and Wolbachia may cooperate as both benefit from ensuring the fitness of the mother. We characterized the relationship between mitochondria and Wolbachia titers in ovaries of Drosophila melanogaster. We found that mitochondria and Wolbachia titers are positively correlated in common laboratory genotypes of D. melanogaster. We attempted to perturb this covariation through the introduction of Wolbachia variants that colonize at different titers. We also attempted to perturb the covariation through manipulating the female reproductive tract to disrupt maternal transmission. Finally, we also attempted to disrupt the covariation by knocking down gene expression for two loci involved in mitochondrial metabolism: NADH dehydrogenase and a mitochondrial transporter. Overall, we find that mitochondria and Wolbachia titers are commonly positively correlated, but this positive covariation is disrupted at high titers of Wolbachia. Our results suggest that mitochondria and Wolbachia have likely evolved mechanisms to stably coexist, but the competitive dynamics change at high Wolbachia titers. We provide future directions to better understand how their interaction influences the maintenance of the symbiosis.}, } @article {pmid29689195, year = {2018}, author = {Foray, V and Pérez-Jiménez, MM and Fattouh, N and Landmann, F}, title = {Wolbachia Control Stem Cell Behavior and Stimulate Germline Proliferation in Filarial Nematodes.}, journal = {Developmental cell}, volume = {45}, number = {2}, pages = {198-211.e3}, doi = {10.1016/j.devcel.2018.03.017}, pmid = {29689195}, issn = {1878-1551}, mesh = {Animals ; Brugia malayi/*growth & development/microbiology ; Cell Proliferation ; Female ; Filariasis/metabolism/parasitology/*pathology ; Germ Cells/*cytology/microbiology/physiology ; Helminth Proteins/genetics/*metabolism ; Male ; Stem Cells/cytology/microbiology/*physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Although symbiotic interactions are ubiquitous in the living world, examples of developmental symbioses are still scarce. We show here the crucial role of Wolbachia in the oogenesis of filarial nematodes, a class of parasites of biomedical and veterinary relevance. We applied newly developed techniques to demonstrate the earliest requirements of Wolbachia in the parasite germline preceding the production of faulty embryos in Wolbachia-depleted nematodes. We show that Wolbachia stimulate germline proliferation in a cell-autonomous manner, and not through nucleotide supplementation as previously hypothesized. We also found Wolbachia to maintain the quiescence of a pool of germline stem cells to ensure a constant delivery of about 1,400 eggs per day for many years. The loss of quiescence upon Wolbachia depletion as well as the disorganization of the distal germline suggest that Wolbachia are required to execute the proper germline stem cell developmental program in order to produce viable eggs and embryos.}, } @article {pmid29686946, year = {2018}, author = {Russell, JE and Nunney, L and Saum, M and Stouthamer, R}, title = {Host and symbiont genetic contributions to fitness in a Trichogramma-Wolbachia symbiosis.}, journal = {PeerJ}, volume = {6}, number = {}, pages = {e4655}, pmid = {29686946}, issn = {2167-8359}, abstract = {The fitness effects associated with Wolbachia infection have wide-ranging ecological and evolutionary consequences for host species. How these effects are modulated by the relative influence of host and Wolbachia genomes has been described as a balancing act of genomic cooperation and conflict. For vertically transmitted symbionts, like cytoplasmic Wolbachia, concordant host-symbiont fitness interests would seem to select for genomic cooperation. However, Wolbachia's ability to manipulate host reproductive systems and distort offspring sex ratios presents an evolutionary conflict of interest with infected hosts. In the parthenogenesis-inducing (PI) form of Wolbachia found in many haplodiploid insects, Wolbachia fitness is realized through females and is enhanced by their feminization of male embryos and subsequent parthenogenetic reproduction. In contrast, as long as Wolbachia is not fixed in a population and sexual reproduction persists, fitness for the host species is realized through both male and female offspring production. How these cooperating and competing interests interact and the relative influence of host and Wolbachia genomes were investigated in the egg parasitoid Trichogramma kaykai, where Wolbachia infection has remained at a low frequency in the field. A factorial design in which laboratory cultures of Wolbachia-infected T. kaykai were cured and re-infected with alternative Wolbachia strains was used to determine the relative influence of host and Wolbachia genomes on host fitness values. Our results suggest fitness variation is largely a function of host genetic background, except in the case of offspring sex ratio where a significant interaction between host and Wolbachia genomes was found. We also find a significant effect associated with the horizontal transfer of Wolbachia strains, which we discuss in terms of the potential for coadaptation in PI-Wolbachia symbioses.}, } @article {pmid29686091, year = {2018}, author = {Shropshire, JD and On, J and Layton, EM and Zhou, H and Bordenstein, SR}, title = {One prophage WO gene rescues cytoplasmic incompatibility in Drosophila melanogaster.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {115}, number = {19}, pages = {4987-4991}, pmid = {29686091}, issn = {1091-6490}, support = {R21 HD086833/HD/NICHD NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; S10 OD021630/OD/NIH HHS/United States ; P30 EY008126/EY/NEI NIH HHS/United States ; P30 DK020593/DK/NIDDK NIH HHS/United States ; P30 CA068485/CA/NCI NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; U24 DK059637/DK/NIDDK NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster ; *Embryo, Nonmammalian/metabolism/microbiology ; Male ; *Prophages ; *Spermatozoa/metabolism/microbiology ; *Wolbachia/genetics/metabolism/virology ; }, abstract = {Wolbachia are maternally inherited, intracellular bacteria at the forefront of vector control efforts to curb arbovirus transmission. In international field trials, the cytoplasmic incompatibility (CI) drive system of wMel Wolbachia is deployed to replace target vector populations, whereby a Wolbachia-induced modification of the sperm genome kills embryos. However, Wolbachia in the embryo rescue the sperm genome impairment, and therefore CI results in a strong fitness advantage for infected females that transmit the bacteria to offspring. The two genes responsible for the wMel-induced sperm modification of CI, cifA and cifB, were recently identified in the eukaryotic association module of prophage WO, but the genetic basis of rescue is unresolved. Here we use transgenic and cytological approaches to demonstrate that maternal cifA expression independently rescues CI and nullifies embryonic death caused by wMel Wolbachia in Drosophila melanogaster Discovery of cifA as the rescue gene and previously one of two CI induction genes establishes a "Two-by-One" model that underpins the genetic basis of CI. Results highlight the central role of prophage WO in shaping Wolbachia phenotypes that are significant to arthropod evolution and vector control.}, } @article {pmid29676724, year = {2018}, author = {Monsanto-Hearne, V and Johnson, KN}, title = {Wolbachia-mediated protection of Drosophila melanogaster against systemic infection with its natural viral pathogen Drosophila C virus does not involve changes in levels of highly abundant miRNAs.}, journal = {The Journal of general virology}, volume = {99}, number = {6}, pages = {827-831}, doi = {10.1099/jgv.0.001064}, pmid = {29676724}, issn = {1465-2099}, mesh = {Animals ; Dicistroviridae/*pathogenicity ; Drosophila melanogaster/*genetics/microbiology/virology ; Host-Pathogen Interactions/*genetics ; MicroRNAs/*genetics ; Real-Time Polymerase Chain Reaction ; Symbiosis ; Virus Diseases/microbiology ; Wolbachia/*physiology ; }, abstract = {The presence of Wolbachia confers virus protection to insects. The molecular mechanism underlying Wolbachia-mediated protection in this tripartite host-endosymbiont-virus interaction is not yet fully understood. In the bipartite association between Drosophila melanogaster and Drosophila C virus (DCV), changes in the expression of microRNAs (miRNAs) influence the outcome of viral pathogenesis. Here we examined whether changes in miRNA expression are similarly involved in the Drosophila-Wolbachia-DCV association. The levels of highly abundant miRNAs in D. melanogaster, Wolbachia-mono-infected D. melanogaster, and DCV- and Wolbachia-bi-infected D. melanogaster were quantified using RT-qPCR and compared. The results show that the abundance of the 17 tested D. melanogaster miRNAs is not affected by Wolbachia endosymbiosis or by bi-infection of Wolbachia and DCV. These results suggest that the in vivo protection conferred by Wolbachia to its native host against D. melanogaster's natural pathogen DCV is not likely to be dependent on or associated with changes in the levels of highly expressed miRNAs.}, } @article {pmid29674896, year = {2018}, author = {Mally, R and Huemer, P and Nuss, M}, title = {Deep intraspecific DNA barcode splits and hybridisation in the Udea alpinalis group (Insecta, Lepidoptera, Crambidae) - an integrative revision.}, journal = {ZooKeys}, volume = {}, number = {746}, pages = {51-90}, pmid = {29674896}, issn = {1313-2989}, abstract = {The analysis of mitochondrial COI data for the European-Centroasian montane Udea alpinalis species group finds deep intraspecific splits. Specimens of U. austriacalis and U. rhododendronalis separate into several biogeographical groups. These allopatric groups are not recovered in the analyses of the two nuclear markers wingless and Elongation factor 1-alpha, except for U. austriacalis from the Pyrenees and the French Massif Central. The latter populations are also morphologically distinct and conspecific with Scopula donzelalis Guenée, 1854, which is removed from synonymy and reinstated as Udea donzelalis (Guenée, 1854) stat. rev. Furthermore, Udea altaica (Zerny, 1914), stat. n. from the Mongolian central Altai mountains, U. juldusalis (Zerny, 1914), stat. n. from the Tian Shan mountains of Kazakhstan, Kyrgyzstan and NW China, and U. plumbalis (Zerny, 1914), stat. n. from the Sayan Mountains of Northern Mongolia are raised to species level, and lectotypes are designated. Evidence of introgression of U. alpinalis into U. uliginosalis at three localities in the Central Alps is presented. A screening for Wolbachia using the markers wsp, gatB and ftsZ was negative for the U. alpinalis species group, but Wolbachia was found in single specimens of U. fulvalis and U. olivalis (both in the U. numeralis species group). We do not find evidence for the conjecture of several authors of additional subspecies in U. rhododendronalis, and synonymise U. rhododendronalis luquetalis Leraut, 1996, syn. n. and U. r. ventosalis Leraut, 1996, syn. n. with the nominal U. rhododendronalis (Duponchel, 1834).}, } @article {pmid29666784, year = {2018}, author = {Ahlers, LRH and Goodman, AG}, title = {The Immune Responses of the Animal Hosts of West Nile Virus: A Comparison of Insects, Birds, and Mammals.}, journal = {Frontiers in cellular and infection microbiology}, volume = {8}, number = {}, pages = {96}, pmid = {29666784}, issn = {2235-2988}, support = {R21 AI128103/AI/NIAID NIH HHS/United States ; T32 GM008336/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Birds/genetics/*immunology/virology ; Humans ; Insecta/genetics/*immunology/virology ; Mammals/genetics/*immunology/virology ; West Nile Fever/genetics/*immunology/*veterinary/virology ; West Nile virus/genetics/*physiology ; }, abstract = {Vector-borne diseases, including arboviruses, pose a serious threat to public health worldwide. Arboviruses of the flavivirus genus, such as Zika virus (ZIKV), dengue virus, yellow fever virus (YFV), and West Nile virus (WNV), are transmitted to humans from insect vectors and can cause serious disease. In 2017, over 2,000 reported cases of WNV virus infection occurred in the United States, with two-thirds of cases classified as neuroinvasive. WNV transmission cycles through two different animal populations: birds and mosquitoes. Mammals, particularly humans and horses, can become infected through mosquito bites and represent dead-end hosts of WNV infection. Because WNV can infect diverse species, research on this arbovirus has investigated the host response in mosquitoes, birds, humans, and horses. With the growing geographical range of the WNV mosquito vector and increased human exposure, improved surveillance and treatment of the infection will enhance public health in areas where WNV is endemic. In this review, we survey the bionomics of mosquito species involved in Nearctic WNV transmission. Subsequently, we describe the known immune response pathways that counter WNV infection in insects, birds, and mammals, as well as the mechanisms known to curb viral infection. Moreover, we discuss the bacterium Wolbachia and its involvement in reducing flavivirus titer in insects. Finally, we highlight the similarities of the known immune pathways and identify potential targets for future studies aimed at improving antiviral therapeutic and vaccination design.}, } @article {pmid29662096, year = {2018}, author = {King, JG and Souto-Maior, C and Sartori, LM and Maciel-de-Freitas, R and Gomes, MGM}, title = {Variation in Wolbachia effects on Aedes mosquitoes as a determinant of invasiveness and vectorial capacity.}, journal = {Nature communications}, volume = {9}, number = {1}, pages = {1483}, pmid = {29662096}, issn = {2041-1723}, mesh = {Aedes/*microbiology ; Animals ; Bayes Theorem ; Brazil/epidemiology ; Dengue/epidemiology/*prevention & control/transmission/virology ; Dengue Virus/growth & development/*pathogenicity ; Disease Susceptibility ; Female ; *Host-Parasite Interactions ; Humans ; *Models, Statistical ; Monte Carlo Method ; Mosquito Vectors/*microbiology ; Vietnam/epidemiology ; Viral Load ; }, abstract = {Wolbachia has been introduced into Aedes aegypti mosquitoes to control the spread of arboviruses, such as dengue, chikungunya and Zika. Studies showed that certain Wolbachia strains (such as wMel) reduce replication of dengue viruses in the laboratory, prompting the release of mosquitoes carrying the bacterium into the field, where vectorial capacity can be realistically assessed in relation to native non-carriers. Here we apply a new analysis to two published datasets, and show that wMel increases the mean and the variance in Ae. aegypti susceptibility to dengue infection when introgressed into Brazil and Vietnam genetic backgrounds. In the absence of other processes, higher mean susceptibility should lead to enhanced viral transmission. The increase in variance, however, widens the basis for selection imposed by unexplored natural forces, retaining the potential for reducing transmission overall.}, } @article {pmid29657018, year = {2018}, author = {Kramer, L and Crosara, S and Gnudi, G and Genchi, M and Mangia, C and Viglietti, A and Quintavalla, C}, title = {Wolbachia, doxycycline and macrocyclic lactones: New prospects in the treatment of canine heartworm disease.}, journal = {Veterinary parasitology}, volume = {254}, number = {}, pages = {95-97}, doi = {10.1016/j.vetpar.2018.03.005}, pmid = {29657018}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Dirofilaria immitis/drug effects ; Dirofilariasis/*drug therapy/parasitology ; Dog Diseases/*drug therapy/parasitology ; Dogs ; Doxycycline/*therapeutic use ; Drug Combinations ; Filaricides/*therapeutic use ; Lactones/*therapeutic use ; Macrocyclic Compounds/therapeutic use ; Wolbachia/*drug effects ; }, abstract = {Melarsomine dihydrochloride (Immiticide®, Merial) is the only approved adulticidal drug for the treatment of canine heartworm disease (HWD). However, in cases where arsenical therapy is not possible or is contraindicated, a monthly heartworm preventive along with doxycycline for a 4-week period, which targets the bacterial endosymbiont Wolbachia, might be considered. There are published reports on the efficacy of ivermectin and doxycycline in both experimentally and naturally infected dogs, but no data on the use of other macrocyclic lactones (MLs) with a similar treatment regime. Preliminary results of studies in dogs show that a topical formulation of moxidectin, the only ML currently registered as a microfilaricide, is also adulticidal when combined with doxycycline. It is not yet known if the efficacy of these combination therapies is due to pharmacokinetic synergism. A recent study showed that serum levels of doxycycline in dogs treated with the combination protocol were not statistically different compared to dogs treated with doxycycline alone. However, lungs from dogs treated with the combination therapy showed a marked reduction in T regulatory cells, indicating that treatment efficacy may be due to a heightened immune response against the parasite. Further studies are necessary to evaluate the long-term clinical outcome of combination protocols and to establish the most efficient treatment for HWD in dogs.}, } @article {pmid29643367, year = {2018}, author = {Bonneau, M and Atyame, C and Beji, M and Justy, F and Cohen-Gonsaud, M and Sicard, M and Weill, M}, title = {Author Correction: Culex pipiens crossing type diversity is governed by an amplified and polymorphic operon of Wolbachia.}, journal = {Nature communications}, volume = {9}, number = {1}, pages = {1491}, doi = {10.1038/s41467-018-03799-4}, pmid = {29643367}, issn = {2041-1723}, abstract = {In the originally published HTML and PDF versions of this Article, gel images in Figures 7c and 8c were not prepared as per the Nature journal policy. These figure panels have now been corrected in both the PDF and HTML versions of the Article.In Fig. 7c, the lane labelled 'Ha' was inappropriately duplicated to represent the lane labelled 'Ich13'. The corrected version of Fig. 7c includes PCR-RFLP on DNA from the Ichkeul 13 line, which had been run on a separate gel. The original unprocessed gel images are provided in Supplementary Figure 1 associated with this correction, with the relevant corresponding bands denoted. A repeat experiment of the PCR-RFLP test is also presented as Supplementary Figure 2.In Fig. 8c, the image was assembled from two separate gels without clear demarcation. The corrected Fig. 8c clearly separates lanes from the two gels, and the original unprocessed gel images are provided in the Supplementary Information associated with this correction.These corrections do not alter the original meaning of the experiments, their results, their interpretation, or the conclusions of the paper. We apologize for any confusion this may have caused to the readers of Nature Communications.}, } @article {pmid29641562, year = {2018}, author = {Asad, S and Hussain, M and Hugo, L and Osei-Amo, S and Zhang, G and Watterson, D and Asgari, S}, title = {Suppression of the pelo protein by Wolbachia and its effect on dengue virus in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {4}, pages = {e0006405}, pmid = {29641562}, issn = {1935-2735}, mesh = {Aedes/genetics/metabolism/*microbiology/*virology ; Animals ; Dengue Virus/*physiology ; Down-Regulation ; Female ; Insect Proteins/*genetics/metabolism ; Insect Vectors/*microbiology/*virology ; MicroRNAs/genetics/metabolism ; Nuclear Proteins/*genetics/metabolism ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {The endosymbiont Wolbachia is known to block replication of several important arboviruses, including dengue virus (DENV), in the mosquito vector Aedes aegypti. So far, the exact mechanism of this viral inhibition is not fully understood. A recent study in Drosophila melanogaster has demonstrated an interaction between the pelo gene and Drosophila C virus. In this study, we explored the possible involvement of the pelo protein, that is involved in protein translation, in Wolbachia-mediated antiviral response and mosquito-DENV interaction. We found that pelo is upregulated during DENV replication and its silencing leads to reduced DENV virion production suggesting that it facilities DENV replication. However, in the presence of Wolbachia, specifically in female mosquitoes, the pelo protein is downregulated and its subcellular localization is altered, which could contribute to reduction in DENV replication in Ae. aegypti. In addition, we show that the microRNA aae-miR-2940-5p, whose abundance is highly enriched in Wolbachia-infected mosquitoes, might mediate regulation of pelo. Our data reveals identification of pelo as a host factor that is positively involved in DENV replication, and its suppression in the presence of Wolbachia may contribute to virus blocking exhibited by the endosymbiont.}, } @article {pmid29636736, year = {2018}, author = {Paniagua Voirol, LR and Frago, E and Kaltenpoth, M and Hilker, M and Fatouros, NE}, title = {Bacterial Symbionts in Lepidoptera: Their Diversity, Transmission, and Impact on the Host.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {556}, pmid = {29636736}, issn = {1664-302X}, abstract = {The insect's microbiota is well acknowledged as a "hidden" player influencing essential insect traits. The gut microbiome of butterflies and moths (Lepidoptera) has been shown to be highly variable between and within species, resulting in a controversy on the functional relevance of gut microbes in this insect order. Here, we aim to (i) review current knowledge on the composition of gut microbial communities across Lepidoptera and (ii) elucidate the drivers of the variability in the lepidopteran gut microbiome and provide an overview on (iii) routes of transfer and (iv) the putative functions of microbes in Lepidoptera. To find out whether Lepidopterans possess a core gut microbiome, we compared studies of the microbiome from 30 lepidopteran species. Gut bacteria of the Enterobacteriaceae, Bacillaceae, and Pseudomonadaceae families were the most widespread across species, with Pseudomonas, Bacillus, Staphylococcus, Enterobacter, and Enterococcus being the most common genera. Several studies indicate that habitat, food plant, and age of the host insect can greatly impact the gut microbiome, which contributes to digestion, detoxification, or defense against natural enemies. We mainly focus on the gut microbiome, but we also include some examples of intracellular endosymbionts. These symbionts are present across a broad range of insect taxa and are known to exert different effects on their host, mostly including nutrition and reproductive manipulation. Only two intracellular bacteria genera (Wolbachia and Spiroplasma) have been reported to colonize reproductive tissues of Lepidoptera, affecting their host's reproduction. We explore routes of transmission of both gut microbiota and intracellular symbionts and have found that these microbes may be horizontally transmitted through the host plant, but also vertically via the egg stage. More detailed knowledge about the functions and plasticity of the microbiome in Lepidoptera may provide novel leads for the control of lepidopteran pest species.}, } @article {pmid29634777, year = {2018}, author = {Pike, A and Dimopoulos, G}, title = {Genetic modification of Anopheles stephensi for resistance to multiple Plasmodium falciparum strains does not influence susceptibility to o'nyong'nyong virus or insecticides, or Wolbachia-mediated resistance to the malaria parasite.}, journal = {PloS one}, volume = {13}, number = {4}, pages = {e0195720}, pmid = {29634777}, issn = {1932-6203}, support = {R01 AI061576/AI/NIAID NIH HHS/United States ; RO1AI061576/NH/NIH HHS/United States ; }, mesh = {Alphavirus Infections/genetics/virology ; Animals ; Animals, Genetically Modified ; Anopheles/*genetics/*parasitology ; Biological Control Agents ; Female ; Host-Parasite Interactions/genetics ; Humans ; Insecticide Resistance/genetics ; Malaria, Falciparum/parasitology/prevention & control/transmission ; Male ; Mosquito Vectors/parasitology ; O'nyong-nyong Virus/pathogenicity ; Plasmodium falciparum/*pathogenicity ; Wolbachia/physiology ; }, abstract = {Mosquitoes that have been genetically engineered for resistance to human pathogens are a potential new tool for controlling vector-borne disease. However, genetic modification may have unintended off-target effects that could affect the mosquitoes' utility for disease control. We measured the resistance of five genetically modified Plasmodium-suppressing Anopheles stephensi lines to o'nyong'nyong virus, four classes of insecticides, and diverse Plasmodium falciparum field isolates and characterized the interactions between our genetic modifications and infection with the bacterium Wolbachia. The genetic modifications did not alter the mosquitoes' resistance to either o'nyong'nyong virus or insecticides, and the mosquitoes were equally resistant to all tested P. falciparum strains, regardless of Wolbachia infection status. These results indicate that mosquitoes can be genetically modified for resistance to malaria parasite infection and remain compatible with other vector-control measures without becoming better vectors for other pathogens.}, } @article {pmid29625205, year = {2018}, author = {Li, Y and Liu, X}, title = {A sex-structured model with birth pulse and release strategy for the spread of Wolbachia in mosquito population.}, journal = {Journal of theoretical biology}, volume = {448}, number = {}, pages = {53-65}, doi = {10.1016/j.jtbi.2018.04.001}, pmid = {29625205}, issn = {1095-8541}, mesh = {Aedes/*microbiology ; Animals ; Computer Simulation ; Dengue/*prevention & control/transmission/virology ; Models, Theoretical ; Mosquito Vectors/microbiology/virology ; Pest Control, Biological/*methods ; Wolbachia/*pathogenicity ; }, abstract = {Dengue fever is one of the most important diseases causing illness and death all over the world, which brings tremendous threat to peoples' life and property security, especially in the undeveloped areas. The main vector, Aedes aegypti, must be controlled to prevent the transmission of dengue. There are a variety of methods to control it. Wolbachia is an innovative bacterium which breaks the dengue transmission cycle for its characteristics of cytoplasmic incompatibility and maternal transmission. In this paper, a sex-structured model with birth pulse is established to study the spread of Wolbachia in mosquito population. The results show that if the maternal transmission is perfect, Wolbachia will spread successfully. Moreover, all the mosquitoes will be infected with Wolbachia. If the maternal transmission is imperfect, there are two locally asymptotically stable periodic solutions. One is Wolbachia-extinction periodic solution, and the other is part replacement periodic solution. Numerical simulations show that the initial occupancy of Wolbachia-infected mosquitoes has an important effect on the success of part replacement strategy. If the initial occupancy is relatively large, the part replacement strategy can be successful. Furthermore, in consideration of the fact that the initial occupancy cannot be always large enough in the wild nature, to release Wolbachia-infected mosquitoes artificially into the wild nature becomes necessary. Therefore, we add a release strategy into the sex-structured model with birth pulse for further analysis. The condition to ensure the stability of the Wolbachia total replacement periodic solution is obtained. Finally, the effect of the release quantity is simulated numerically.}, } @article {pmid29618379, year = {2018}, author = {Guo, Y and Song, Z and Luo, L and Wang, Q and Zhou, G and Yang, D and Zhong, D and Zheng, X}, title = {Molecular evidence for new sympatric cryptic species of Aedes albopictus (Diptera: Culicidae) in China: A new threat from Aedes albopictus subgroup?.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {228}, pmid = {29618379}, issn = {1756-3305}, support = {31630011//National Natural Science Foundation of China (CN)/International ; 2013B021800042//Science and Technology Plan Project of Guangdong Province (CN)/International ; 2015A030313784//Natural Science Foundation of Guangdong Province (CN)/International ; }, mesh = {Aedes/*classification/genetics/*growth & development/microbiology ; Animals ; China ; Cluster Analysis ; DNA, Bacterial/analysis/genetics ; DNA, Ribosomal Spacer/chemistry/genetics ; Electron Transport Complex IV/genetics ; *Genetic Variation ; *Genotype ; Mosquito Vectors/*classification/genetics/*growth & development ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/genetics/isolation & purification ; }, abstract = {BACKGROUND: Aedes (Stegomyia) albopictus (Skuse) is an indigenous species and the predominant vector of dengue fever in China. Understanding of genetic diversity and structure of the mosquito would facilitate dengue prevention and vector control. Sympatric cryptic species have been identified in the Ae. albopictus subgroup in Southeast Asia; however, little is known about the presence and distribution of cryptic species in China. This study aimed to examine the genetic diversity, evaluate potential new cryptic sibling species, and assess the prevalence of Wolbachia infections in field populations.

METHODS: Aedes adult female specimens were collected from five provinces in southern and central China during 2015-2016. Morphological identification was performed under dissection microscope. The mitochondrial DNA cytochrome c oxidase subunit 1 (cox1, DNA barcoding) locus and the ribosomal DNA internal transcribed spacer region 2 (ITS2) marker were used to examine the genetic variation, evaluate cryptic sibling species, and population structure in the field populations. Screening for the presence of Wolbachia was performed using multiplex PCR.

RESULTS: A total of 140 individual specimens with morphological characteristics similar to Ae. albopictus were sequenced for DNA barcoding. Among these, 129 specimens (92.1%) were confirmed and identified as Ae. albopictus. The remaining 11 specimens, from 2 provinces, were identified as 2 distinct sequence groups, which were confirmed by ITS2 marker sequencing, suggesting the existence of potential cryptic species of Ae. albopictus. In Ae. albopictus, we found significant genetic differentiation and population structure between populations collected from different climate zones. Medium to high frequencies of Wolbachia infections were observed in natural Ae. albopictus populations, whereas Wolbachia was infrequent or absent in cryptic species populations.

CONCLUSIONS: Our findings highlight the population differentiation by climate zone and the presence of novel, cryptic Aedes species in China. The low prevalence of Wolbachia infections in cryptic species populations could reflect either a recent invasion of Wolbachia in Ae. albopictus or different host immune responses to this symbiont in the cryptic species. The study provides useful information for vector control and host-symbiont coevolution. Further study is needed to investigate the potential for arbovirus infection and disease transmission in the emerged cryptic species.}, } @article {pmid29607029, year = {2018}, author = {Fortin, M and Debenest, C and Souty-Grosset, C and Richard, FJ}, title = {Males prefer virgin females, even if parasitized, in the terrestrial isopod Armadillidium vulgare.}, journal = {Ecology and evolution}, volume = {8}, number = {6}, pages = {3341-3353}, pmid = {29607029}, issn = {2045-7758}, abstract = {In many species, males increase their reproductive success by choosing high-quality females. In natural populations, they interact with both virgin and mated females, which can store sperm in their spermatheca. Therefore, males elaborate strategies to avoid sperm competition. In the terrestrial isopod Armadillidium vulgare, females can store sperm and produce several clutches. Moreover, this species can be parasitized by Wolbachia, which feminizes genetic males, transforming them into functional females. Our study compared attractiveness and mate choice when a male is exposed to both virgin and experienced females (i.e., females who have produced offspring and rested for 6 months), with or without Wolbachia. Our results revealed that males are more attracted to virgin females than experienced females, even if these virgin females are parasitized. Moreover, the chemical analysis highlighted different odors in females according to their reproductive and infection (Wolbachia-free or vertically Wolbachia-infected) status. Males attempted copulation more frequently and for longer with virgin females, even if Wolbachia-infected, while experienced females refused further copulation. The evolutionary consequences of both male choice and female resistance on their fitness are discussed in this study.}, } @article {pmid29603499, year = {2018}, author = {Mariño, YA and Ospina, OE and Verle Rodrigues, JC and Bayman, P}, title = {High diversity and variability in the bacterial microbiota of the coffee berry borer (Coleoptera: Curculionidae), with emphasis on Wolbachia.}, journal = {Journal of applied microbiology}, volume = {125}, number = {2}, pages = {528-543}, doi = {10.1111/jam.13768}, pmid = {29603499}, issn = {1365-2672}, mesh = {Animals ; Coffea/*parasitology ; DNA, Bacterial/analysis/genetics ; Microbiota/*genetics ; Weevils/*microbiology ; Wolbachia/*genetics ; }, abstract = {AIMS: Variation in microbiota of the coffee berry borer (CBB) Hypothenemus hampei was studied. Diversity, structure and function of bacterial communities were compared between eggs vs adults, CBBs from shade coffee vs sun coffee, CBBs from the field vs raised in the laboratory, and CBBs with and without the antibiotic tetracycline.

METHODS AND RESULTS: We sequenced the region V4 of the gene 16 S rRNA. Pseudomonadaceae and Enterobacteriaceae, particularly Pseudomonas and Pantoea, dominated microbiotas of the CBB. Comparative functional inferences with PICRUSt suggested that samples from the field were enriched for genes involved in carbohydrate and protein digestion and absorption, while laboratory-reared samples were higher in genes for melanization and caffeine metabolism.

CONCLUSIONS: Microbiotas of the CBB were diverse and dominated by the genus Pseudomonas, several species of which have been previously associated with caffeine degradation in this insect. Wolbachia was the only endosymbiont detected with known ability to manipulate host reproduction.

This study demonstrates that stage of development and origin of samples affected the structure and function of the CBB's bacterial communities. This is the first attempt to predict functional significance of the CBB microbiota in nutrition, reproduction and defence.}, } @article {pmid29596879, year = {2018}, author = {Ali, H and Muhammad, A and Islam, SU and Islam, W and Hou, Y}, title = {A novel bacterial symbiont association in the hispid beetle, Octodonta nipae (Coleoptera: Chrysomelidae), their dynamics and phylogeny.}, journal = {Microbial pathogenesis}, volume = {118}, number = {}, pages = {378-386}, doi = {10.1016/j.micpath.2018.03.046}, pmid = {29596879}, issn = {1096-1208}, mesh = {Animals ; *Bacterial Physiological Phenomena ; Bacterial Proteins/genetics ; Coleoptera/*microbiology/pathogenicity/physiology ; DNA, Bacterial/analysis ; Female ; Genes, Bacterial/genetics ; Genetic Vectors ; Genome Size ; Genotyping Techniques ; Larva/microbiology ; Male ; Multilocus Sequence Typing ; *Phylogeny ; Plant Diseases/parasitology ; Reproduction ; Sequence Alignment ; *Symbiosis ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {The hispid leaf beetle, Octodonta nipae (Maulik), (Coleoptera: Chrysomelidae), is a devastating pest of palm cultivation worldwide. Endosymbiotic bacteria in the genus Wolbachia are arguably one of the most abundant bacterial group associated with arthropods. Owing to its critical effects on host reproduction, Wolbachia has garnered much attention as a prospective future tool for insect pest management. However, their association, infection dynamics, and functionality remain unknown in this insect pest. Here, we diagnosis for the first time, the infection prevalence, and occurrence of Wolbachia in O. nipae. Experimental evidence by the exploration of wsp gene vindicate that O. nipae is naturally infected with bacterial symbiont of genus Wolbachia, showing a complete maternal inheritance with shared a common Wolbachia strain (wNip). Moreover, MLST (gatB, fbpA, coxA, ftsZ, and hcpA) analysis enabled the detections of new sequence type (ST-484), suggesting a particular genotypic association of O. nipae and Wolbachia. Subsequently, quantitative real-time PCR (qPCR) assay demonstrated variable infection density across different life stages (eggs, larvae, pupae and adult male and female), body parts (head, thorax, abdomen), and tissues (ovaries, testes, and guts). Infection density was higher in egg and female adult stage, as well as abdomen and reproductive tissues as compared to other samples. Interestingly, Wolbachia harbored dominantly in a female than the male adult, while, no significant differences were observed between male and female body parts and tissues. Phylogeny of Wolbachia infection associated with O. nipae rectified from all tested life stages were unique and fall within the same monophyletic supergroup-A of Wolbachia clades. The infection density of symbiont is among the valuable tool to understand their biological influence on hosts, and this latest discovery would facilitate the future investigations to understand the host-symbiont complications and its prospective role as a microbiological agent to reduce pest populations.}, } @article {pmid29593683, year = {2018}, author = {Di Guardo, G}, title = {Commentary: Zika Virus in the Americas-Yet Another Arbovirus Threat.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {435}, pmid = {29593683}, issn = {1664-302X}, } @article {pmid29587626, year = {2018}, author = {Schuler, H and Egan, SP and Hood, GR and Busbee, RW and Driscoe, AL and Ott, JR}, title = {Diversity and distribution of Wolbachia in relation to geography, host plant affiliation and life cycle of a heterogonic gall wasp.}, journal = {BMC evolutionary biology}, volume = {18}, number = {1}, pages = {37}, pmid = {29587626}, issn = {1471-2148}, support = {J-3527-B22//Austrian Science Fund/International ; }, mesh = {Animals ; Bayes Theorem ; DNA, Mitochondrial/genetics ; Electron Transport Complex IV/genetics ; Female ; *Genetic Variation ; Genetics, Population ; *Geography ; Haplotypes/genetics ; *Life Cycle Stages ; Male ; Phylogeny ; Quercus/*parasitology ; United States ; Wasps/*genetics/*microbiology ; Wolbachia/genetics/*growth & development ; }, abstract = {BACKGROUND: The maternally inherited endosymbiont Wolbachia is widespread in arthropods and nematodes and can play an important role in the ecology and evolution of its host through reproductive manipulation. Here, we survey Wolbachia in Belonocnema treatae, a widely distributed North American cynipid gall forming wasp that exhibits regional host specialization on three species of oaks and alternation of sexually and asexually reproducing generations. We investigated whether patterns of Wolbachia infection and diversity in B. treatae are associated with the insect's geographic distribution, host plant association, life cycle, and mitochondrial evolutionary history.

RESULTS: Screening of 463 individuals from 23 populations including sexual and asexual generations from all three host plants across the southern U.S. showed an average infection rate of 56% with three common Wolbachia strains: wTre1-3 and an additional rare variant wTre4. Phylogenetic analysis based on wsp showed that these strains are unrelated and likely independently inherited. We found no difference in Wolbachia infection frequency among host plant associated populations or between the asexual and sexual generations, or between males and females of the sexual generation. Partially incomplete Wolbachia transmission rates might explain the occurrence of uninfected individuals. A parallel analysis of the mitochondrial cytochrome oxidase I gene in B. treatae showed high mtDNA haplotype diversity in both infected and uninfected populations suggesting an ancestral infection by Wolbachia as well as a clear split between eastern and western B. treatae mtDNA clades with a sequence divergence of > 6%. The strain wTre1 was present almost exclusively in the western clade while wTre2 and wTre3 occur almost exclusively in eastern populations. In contrast, the same strains co-occur as double-infections in Georgia and triple-infections in two populations in central Florida.

CONCLUSIONS: The diversity of Wolbachia across geographically and genetically distinct populations of B. treatae and the co-occurrence of the same strains within three populations highlights the complex infection dynamics in this system. Moreover, the association of distinct Wolbachia strains with mitochondrial haplotypes of its host in populations infected by different Wolbachia strains suggests a potential role of the endosymbiont in reproductive isolation in B. treatae.}, } @article {pmid29579215, year = {2018}, author = {Diouf, M and Miambi, E and Mora, P and Frechault, S and Robert, A and Rouland-Lefèvre, C and Hervé, V}, title = {Variations in the relative abundance of Wolbachia in the gut of Nasutitermes arborum across life stages and castes.}, journal = {FEMS microbiology letters}, volume = {365}, number = {7}, pages = {}, doi = {10.1093/femsle/fny046}, pmid = {29579215}, issn = {1574-6968}, mesh = {Animals ; Bacteria/classification/genetics/growth & development/isolation & purification ; *Gastrointestinal Microbiome ; Gastrointestinal Tract/microbiology ; Isoptera/*growth & development/*microbiology ; Larva/growth & development/microbiology ; Life Cycle Stages ; Phylogeny ; Wolbachia/classification/genetics/growth & development/*isolation & purification ; }, abstract = {There are multiple forms of interactions between termites and bacteria. In addition to their gut microbiota, which has been intensively studied, termites host intracellular symbionts such as Wolbachia. These distinct symbioses have been so far approached independently and mostly in adult termites. We addressed the dynamics of Wolbachia and the microbiota of the eggs and gut for various life stages and castes of the wood-feeding termite, Nasutitermes arborum, using deep-sequencing of the 16S rRNA gene. Wolbachia was dominant in eggs as expected. Unexpectedly, it persisted in the gut of nearly all stages and castes, indicating a wide somatic distribution in termites. Wolbachia-related sequences clustered into few operational taxonomic units, but these were within the same genotype, acquired maternally. Wolbachia was largely dominant in DNA extracts from the guts of larvae and pre-soldiers (59.1%-99.1% of reads) where gut-resident lineages were less represented and less diverse. The reverse was true for the adult castes. This is the first study reporting the age-dependency of the relative abundance of Wolbachia in the termite gut and its negative correlation with the diversity of the microbiota. The possible mechanisms underlying this negative interaction are discussed.}, } @article {pmid29573202, year = {2018}, author = {Ren, SL and Li, YH and Ou, D and Guo, YJ and Qureshi, JA and Stansly, PA and Qiu, BL}, title = {Localization and dynamics of Wolbachia infection in Asian citrus psyllid Diaphorina citri, the insect vector of the causal pathogens of Huanglongbing.}, journal = {MicrobiologyOpen}, volume = {7}, number = {3}, pages = {e00561}, pmid = {29573202}, issn = {2045-8827}, mesh = {Animal Structures/microbiology ; Animals ; Asia ; Bacterial Load ; Female ; Hemiptera/*growth & development/*microbiology ; In Situ Hybridization, Fluorescence ; Insect Vectors/*microbiology ; Male ; Nymphaea/microbiology ; Wolbachia/*isolation & purification ; Zygote/microbiology ; }, abstract = {Wolbachia is a group of intracellular bacteria that infect a wide range of arthropods including the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. This insect is the vector of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of Huanglongbing or citrus greening disease. Here, we investigated the localization pattern and infection dynamics of Wolbachia in different developmental stages of ACP. Results revealed that all developmental stages of ACP including egg, 1st-5th instar nymphs, and adults of both gender were infected with Wolbachia. FISH visualization of an ACP egg showed that Wolbachia moved from the egg stalk of newly laid eggs to a randomly distributed pattern throughout the egg prior to hatching. The infection rate varied between nymphal instars. The titers of Wolbachia in fourth and fifth instar nymphs were significantly higher than those in the first and second instar nymphs. Wolbachia were scattered in all nymphal stages, but with highest intensity in the U-shaped bacteriome located in the abdomen of the nymph. Wolbachia was confined to two symmetrical organizations in the abdomen of newly emerged female and male adults. The potential mechanisms of Wolbachia infection dynamics are discussed.}, } @article {pmid29569277, year = {2018}, author = {Dincă, V and Bálint, Z and Vodă, R and Dapporto, L and Hebert, PDN and Vila, R}, title = {Use of genetic, climatic, and microbiological data to inform reintroduction of a regionally extinct butterfly.}, journal = {Conservation biology : the journal of the Society for Conservation Biology}, volume = {32}, number = {4}, pages = {828-837}, doi = {10.1111/cobi.13111}, pmid = {29569277}, issn = {1523-1739}, mesh = {Animals ; *Butterflies ; Conservation of Natural Resources ; DNA, Mitochondrial ; France ; Genetic Variation ; Haplotypes ; Phylogeny ; }, abstract = {Species reintroductions are increasingly used as means of mitigating biodiversity loss. Besides habitat quality at the site targeted for reintroduction, the choice of source population can be critical for success. The butterfly Melanargia russiae (Esper´s marbled white) was extirpated from Hungary over 100 years ago, and a reintroduction program has recently been approved. We used museum specimens of this butterfly, mitochondrial DNA data (mtDNA), endosymbiont screening, and climatic-similarity analyses to determine which extant populations should be used for its reintroduction. The species displayed 2 main mtDNA lineages across its range: 1 restricted to Iberia and southern France (Iberian lineage) and another found throughout the rest of its range (Eurasian lineage). These 2 lineages possessed highly divergent wsp alleles of the bacterial endosymbiont Wolbachia. The century-old Hungarian specimens represented an endemic haplotype belonging to the Eurasian lineage, differing by one mutation from the Balkan and eastern European populations. The Hungarian populations of M. russiae occurred in areas with a colder and drier climate relative to most sites with extant known populations. Our results suggest the populations used for reintroduction to Hungary should belong to the Eurasian lineage, preferably from eastern Ukraine (genetically close and living in areas with the highest climatic similarity). Materials stored in museum collections can provide unique opportunities to document historical genetic diversity and help direct conservation.}, } @article {pmid29569154, year = {2018}, author = {Li, Y and Liu, X and Guo, H}, title = {Variations in Endosymbiont Infection Between Buprofezin-Resistant and Susceptible Strains of Laodelphax striatellus (Fallén).}, journal = {Current microbiology}, volume = {75}, number = {6}, pages = {709-715}, pmid = {29569154}, issn = {1432-0991}, support = {31672027//The National Natural Science Foundation of China/ ; cx(16)1001//The Independent Innovation Fund of Agricultural Science and Technology in Jiangsu province, China/ ; }, mesh = {Serratia/drug effects/metabolism/*physiology ; Thiadiazines/*pharmacology ; Wolbachia/drug effects/metabolism/*physiology ; }, abstract = {The endosymbionts Wolbachia and Rickettsia have been shown to be correlated with the insecticide resistance of mosquito and whitefly. The small brown planthopper (SBPH), Laodelphax striatellus, harbours many species of endosymbionts, and has developed a high resistance to buprofezin in China. In this study, we examined the species and the infection incidences of endosymbionts in a buprofezin-resistant (BR) strain, a buprofezin-susceptible (BS) strain, and the BR strain after exposure to buprofezin, and we also investigated the change in buprofezin susceptibility after removal of Wolbachia from the BR strain. The results showed that Wolbachia infection incidences were 100% in both the BR and BS strains, but the Wolbachia density in the BR strain was significantly higher than that in the BS strain. There were no significant differences in Arsenophonus infection incidence between the two strains. However, the infection incidence of Serratia and double infection incidence of Serratia + Wolbachia in the BR strain were significantly higher than that in the BS strain. After the BR strain was exposed to 1200 mg/L buprofezin, the infection incidence of Arsenophonus in the surviving individuals increased, and the infection rate of Serratia did not differ, but the double infection incidence of Serratia + Wolbachia decreased. And when a Wolbachia-infected line originating from the BR strain was cleared of Wolbachia, its susceptibility to buprofezin increased. The results suggest that Serratia and Wolbachia infection might improve the buprofezin resistance of SBPH.}, } @article {pmid29568706, year = {2018}, author = {Kajtoch, Ł and Kotásková, N}, title = {Current state of knowledge on Wolbachia infection among Coleoptera: a systematic review.}, journal = {PeerJ}, volume = {6}, number = {}, pages = {e4471}, pmid = {29568706}, issn = {2167-8359}, abstract = {BACKGROUND: Despite great progress in studies on Wolbachia infection in insects, the knowledge about its relations with beetle species, populations and individuals, and the effects of bacteria on these hosts, is still unsatisfactory. In this review we summarize the current state of knowledge about Wolbachia occurrence and interactions with Coleopteran hosts.

METHODS: An intensive search of the available literature resulted in the selection of 86 publications that describe the relevant details about Wolbachia presence among beetles. These publications were then examined with respect to the distribution and taxonomy of infected hosts and diversity of Wolbachia found in beetles. Sequences of Wolbachia genes (16S rDNA, ftsZ) were used for the phylogenetic analyses.

RESULTS: The collected publications revealed that Wolbachia has been confirmed in 204 beetle species and that the estimated average prevalence of this bacteria across beetle species is 38.3% and varies greatly across families and genera (0-88% infected members) and is much lower (c. 13%) in geographic studies. The majority of the examined and infected beetles were from Europe and East Asia. The most intensively studied have been two groups of herbivorous beetles: Curculionidae and Chrysomelidae. Coleoptera harbor Wolbachia belonging to three supergroups: F found in only three species, and A and B found in similar numbers of beetles (including some doubly infected); however the latter two were most prevalent in different families. A total of 59% of species with precise data were found to be totally infected. Single infections were found in 69% of species and others were doubly- or multiply-infected. Wolbachia caused numerous effects on its beetle hosts, including selective sweep with host mtDNA (found in 3% of species), cytoplasmic incompatibility (detected in c. 6% of beetles) and other effects related to reproduction or development (like male-killing, possible parthenogenesis or haplodiploidy induction, and egg development). Phylogenetic reconstructions for Wolbachia genes rejected cospeciation between these bacteria and Coleoptera, with minor exceptions found in some Hydraenidae, Curculionidae and Chrysomelidae. In contrast, horizontal transmission of bacteria has been suspected or proven in numerous cases (e.g., among beetles sharing habitats and/or host plants).

DISCUSSION: The present knowledge about Wolbachia infection across beetle species and populations is very uneven. Even the basic data about infection status in species and frequency of infected species across genera and families is very superficial, as only c. 0.15% of all beetle species have been tested so far. Future studies on Wolbachia diversity in Coleoptera should still be based on the Multi-locus Sequence Typing system, and next-generation sequencing technologies will be important for uncovering Wolbachia relations with host evolution and ecology, as well as with other, co-occurring endosymbiotic bacteria.}, } @article {pmid29568248, year = {2018}, author = {Nikolouli, K and Colinet, H and Renault, D and Enriquez, T and Mouton, L and Gibert, P and Sassu, F and Cáceres, C and Stauffer, C and Pereira, R and Bourtzis, K}, title = {Sterile insect technique and Wolbachia symbiosis as potential tools for the control of the invasive species Drosophila suzukii.}, journal = {Journal of pest science}, volume = {91}, number = {2}, pages = {489-503}, pmid = {29568248}, issn = {1612-4758}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Drosophila suzukii, a vinegar fly originated from Southeast Asia, has recently invaded western countries, and it has been recognized as an important threat of a wide variety of several commercial soft fruits. This review summarizes the current information about the biology and dispersal of D. suzukii and discusses the current status and prospects of control methods for the management of this pest. We highlight current knowledge and ongoing research on innovative environmental-friendly control methods with emphasis on the sterile insect technique (SIT) and the incompatible insect technique (IIT). SIT has been successfully used for the containment, suppression or even eradication of populations of insect pests. IIT has been proposed as a stand-alone tool or in conjunction with SIT for insect pest control. The principles of SIT and IIT are reviewed, and the potential value of each approach in the management of D. suzukii is analyzed. We thoroughly address the challenges of SIT and IIT, and we propose the use of SIT as a component of an area-wide integrated pest management approach to suppress D. suzukii populations. As a contingency plan, we suggest a promising alternative avenue through the combination of these two techniques, SIT/IIT, which has been developed and is currently being tested in open-field trials against Aedes mosquito populations. All the potential limiting factors that may render these methods ineffective, as well as the requirements that need to be fulfilled before their application, are discussed.}, } @article {pmid29568046, year = {2018}, author = {Chuchuy, A and Rodriguero, MS and Ferrari, W and Ciota, AT and Kramer, LD and Micieli, MV}, title = {Biological characterization of Aedes albopictus (Diptera: Culicidae) in Argentina: implications for arbovirus transmission.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {5041}, pmid = {29568046}, issn = {2045-2322}, mesh = {Aedes/microbiology/pathogenicity/*virology ; Animals ; Arbovirus Infections/epidemiology/microbiology/*transmission/virology ; Arboviruses/*genetics/pathogenicity ; Argentina/epidemiology ; Asia/epidemiology ; Disease Vectors ; Female ; Humans ; Male ; Mosquito Vectors/microbiology/*physiology/virology ; Wolbachia/pathogenicity ; }, abstract = {Aedes albopictus (Diptera: Culicidae) is an invasive mosquito, native to Asia, that has expanded its range worldwide. It is considered to be a public health threat as it is a competent vector of viruses of medical importance, including dengue, chikungunya, and Zika. Despite its medical importance there is almost no information on biologically important traits of Ae. albopictus in Argentina. We studied life cycle traits, demographic parameters and analyzed the competence of this mosquito as a virus vector. In addition, we determined the prevalence of Wolbachia strains in Ae. albopictus as a first approach to investigate the potential role of this bacteria in modulating vector competence for arboviruses. We observed low hatch rates of eggs, which led to a negative growth rate. We found that Ae. albopictus individuals were infected with Wolbachia in the F1 but while standard superinfection with wAlbA and wAlbB types was found in 66.7% of the females, 16.7% of the females and 62.5% of the males were single-infected with the wAlbB strain. Finally, despite high levels of infection and dissemination, particularly for chikungunya virus, Ae. albopictus from subtropical Argentina were found to be relatively inefficient vectors for transmission of both chikungunya and dengue viruses.}, } @article {pmid29563258, year = {2018}, author = {Harumoto, T and Fukatsu, T and Lemaitre, B}, title = {Common and unique strategies of male killing evolved in two distinct Drosophila symbionts.}, journal = {Proceedings. Biological sciences}, volume = {285}, number = {1875}, pages = {}, pmid = {29563258}, issn = {1471-2954}, support = {339970/ERC_/European Research Council/International ; }, mesh = {Animals ; Apoptosis ; DNA Damage ; Dosage Compensation, Genetic ; Drosophila/*embryology/genetics/*microbiology ; Embryonic Development ; Female ; In Situ Nick-End Labeling ; Male ; Nervous System/microbiology ; Sex Factors ; Spiroplasma/*growth & development/pathogenicity ; *Symbiosis ; Wolbachia/*growth & development/pathogenicity ; }, abstract = {Male killing is a selfish reproductive manipulation caused by symbiotic bacteria, where male offspring of infected hosts are selectively killed. The underlying mechanisms and the process of their evolution are of great interest not only in terms of fundamental biology, but also their potential applications. The two bacterial Drosophila symbionts, Wolbachia and Spiroplasma, have independently evolved male-killing ability. This raises the question whether the underlying mechanisms share some similarities or are specific to each bacterial species. Here, we analyse pathogenic phenotypes of D. bifasciata infected with its natural male-killing Wolbachia strain and compare them with those of D. melanogaster infected with male-killing Spiroplasma We show that male progeny infected with the Wolbachia strain die during embryogenesis with abnormal apoptosis. Interestingly, male-killing Wolbachia infection induces DNA damage and segregation defects in the dosage-compensated chromosome in male embryos, which are reminiscent of the phenotypes caused by male-killing Spiroplasma in D. melanogaster By contrast, host neural development seems to proceed normally unlike male-killing Spiroplasma infection. Our results demonstrate that the dosage-compensated chromosome is a common target of two distinct male killers, yet Spiroplasma uniquely evolved the ability to damage neural tissue of male embryos.}, } @article {pmid29561780, year = {2018}, author = {Lindsey, ARI and Bhattacharya, T and Newton, ILG and Hardy, RW}, title = {Conflict in the Intracellular Lives of Endosymbionts and Viruses: A Mechanistic Look at Wolbachia-Mediated Pathogen-blocking.}, journal = {Viruses}, volume = {10}, number = {4}, pages = {}, pmid = {29561780}, issn = {1999-4915}, support = {R21 AI121849/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Antibiosis ; Biological Transport ; *Coinfection ; Disease Resistance/genetics/immunology ; Genotype ; *Host-Pathogen Interactions ; Humans ; Insecta/microbiology/virology ; Intracellular Space/microbiology/virology ; Protein Biosynthesis ; RNA Interference ; Rickettsiaceae Infections/*microbiology ; Stress, Physiological ; *Symbiosis ; Virulence ; Virus Assembly ; Virus Diseases/*virology ; Virus Internalization ; *Virus Physiological Phenomena ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {At the forefront of vector control efforts are strategies that leverage host-microbe associations to reduce vectorial capacity. The most promising of these efforts employs Wolbachia, a maternally transmitted endosymbiotic bacterium naturally found in 40% of insects. Wolbachia can spread through a population of insects while simultaneously inhibiting the replication of viruses within its host. Despite successes in using Wolbachia-transfected mosquitoes to limit dengue, Zika, and chikungunya transmission, the mechanisms behind pathogen-blocking have not been fully characterized. Firstly, we discuss how Wolbachia and viruses both require specific host-derived structures, compounds, and processes to initiate and maintain infection. There is significant overlap in these requirements, and infection with either microbe often manifests as cellular stress, which may be a key component of Wolbachia's anti-viral effect. Secondly, we discuss the current understanding of pathogen-blocking through this lens of cellular stress and develop a comprehensive view of how the lives of Wolbachia and viruses are fundamentally in conflict with each other. A thorough understanding of the genetic and cellular determinants of pathogen-blocking will significantly enhance the ability of vector control programs to deploy and maintain effective Wolbachia-mediated control measures.}, } @article {pmid29558974, year = {2018}, author = {Altinli, M and Gunay, F and Alten, B and Weill, M and Sicard, M}, title = {Wolbachia diversity and cytoplasmic incompatibility patterns in Culex pipiens populations in Turkey.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {198}, pmid = {29558974}, issn = {1756-3305}, support = {ANR-16-CE02-0006-01//Agence Nationale de la Recherche/International ; }, mesh = {Animals ; Culex/*microbiology ; Cytoplasm/*microbiology/pathology ; DNA, Mitochondrial/genetics ; *Disease Vectors ; *Genetic Variation ; Gram-Negative Bacterial Infections/epidemiology ; Mosquito Vectors/*microbiology ; Polymerase Chain Reaction ; Reproduction ; Symbiosis ; Turkey/epidemiology ; Wolbachia/*genetics/isolation & purification/physiology ; }, abstract = {BACKGROUND: Wolbachia are maternally transmitted bacteria that can manipulate their hosts' reproduction causing cytoplasmic incompatibility (CI). CI is a sperm-egg incompatibility resulting in embryonic death. Due to this sterilising effect on mosquitoes, Wolbachia are considered for vector control strategies. Important vectors for arboviruses, filarial nematodes and avian malaria, mosquitoes of Culex pipiens complex are suitable for Wolbachia-based vector control. They are infected with Wolbachia wPip strains belonging to five genetically distinct groups (wPip-I to V) within the Wolbachia B supergroup. CI properties of wPip strongly correlate with this genetic diversity: mosquitoes infected with wPip strains from a different wPip group are more likely to be incompatible with each other. Turkey is a critical spot for vector-borne diseases due to its unique geographical position as a natural bridge between Asia, Europe and Africa. However, general wPip diversity, distribution and CI patterns in natural Cx. pipiens (s.l.) populations in the region are unknown. In this study, we first identified wPip diversity in Turkish Cx. pipiens (s.l.) populations, by assigning them to one of the five groups within wPip (wPip-Ito V). We further investigated CI properties between different wPip strains from this region.

RESULTS: We showed a wPip fixation in Cx. pipiens (s.l.) populations in Turkey by analysing 753 samples from 59 sampling sites. Three wPip groups were detected in the region: wPip-I, wPip-II and wPip-IV. The most dominant group was wPip-II. While wPip-IV was restricted to only two locations, wPip-I and wPip-II had wider distributions. Individuals infected with wPip-II were found co-existing with individuals infected with wPip-I or wPip-IV in some sampling sites. Two mosquito isofemale lines harbouring either a wPip-I or a wPip-II strain were established from a population in northwestern Turkey. Reciprocal crosses between these lines showed that they were fully compatible with each other but bidirectionally incompatible with wPip-IV Istanbul infected line.

CONCLUSION: Our findings reveal a high diversity of wPip and CI properties in Cx. pipiens (s.l.) populations in Turkey. Knowledge on naturally occurring CI patterns caused by wPip diversity in Turkey might be useful for Cx. pipiens (s.l.) control in the region.}, } @article {pmid29558644, year = {2018}, author = {Siozios, S and Gerth, M and Griffin, JS and Hurst, GDD}, title = {Symbiosis: Wolbachia Host Shifts in the Fast Lane.}, journal = {Current biology : CB}, volume = {28}, number = {6}, pages = {R269-R271}, doi = {10.1016/j.cub.2018.02.008}, pmid = {29558644}, issn = {1879-0445}, mesh = {Animals ; Arthropods/*microbiology ; Drosophila ; Symbiosis ; *Wolbachia ; }, abstract = {The inherited bacterium Wolbachia is an important component of the biology of many arthropods. What makes it so common? An analysis of drosophilids revealed one strain host shifts at a surprisingly high rate, infecting eight species in under 30,000 years.}, } @article {pmid29558464, year = {2018}, author = {Souto-Maior, C and Sylvestre, G and Braga Stehling Dias, F and Gomes, MGM and Maciel-de-Freitas, R}, title = {Model-based inference from multiple dose, time course data reveals Wolbachia effects on infection profiles of type 1 dengue virus in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {3}, pages = {e0006339}, pmid = {29558464}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*virology ; Animals ; Dengue/prevention & control/transmission ; Dengue Virus/*physiology ; Host-Pathogen Interactions ; *Models, Biological ; Mosquito Vectors/*microbiology/*virology ; Symbiosis ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Infection is a complex and dynamic process involving a population of invading microbes, the host and its responses, aimed at controlling the situation. Depending on the purpose and level of organization, infection at the organism level can be described by a process as simple as a coin toss, or as complex as a multi-factorial dynamic model; the former, for instance, may be adequate as a component of a population model, while the latter is necessary for a thorough description of the process beginning with a challenge with an infectious inoculum up to establishment or elimination of the pathogen. Experimental readouts in the laboratory are often static, snapshots of the process, assayed under some convenient experimental condition, and therefore cannot comprehensively describe the system. Different from the discrete treatment of infection in population models, or the descriptive summarized accounts of typical lab experiments, in this manuscript, infection is treated as a dynamic process dependent on the initial conditions of the infectious challenge, viral growth, and the host response along time. Here, experimental data is generated for multiple doses of type 1 dengue virus, and pathogen levels are recorded at different points in time for two populations of mosquitoes: either carrying endosymbiont bacteria Wolbachia or not. A dynamic microbe/host-response mathematical model is used to describe pathogen growth in the face of a host response like the immune system, and to infer model parameters for the two populations of insects, revealing a slight-but potentially important-protection conferred by the symbiont.}, } @article {pmid29530790, year = {2018}, author = {Zhang, X and Tang, S and Liu, Q and Cheke, RA and Zhu, H}, title = {Models to assess the effects of non-identical sex ratio augmentations of Wolbachia-carrying mosquitoes on the control of dengue disease.}, journal = {Mathematical biosciences}, volume = {299}, number = {}, pages = {58-72}, doi = {10.1016/j.mbs.2018.03.003}, pmid = {29530790}, issn = {1879-3134}, support = {//CIHR/Canada ; }, mesh = {Aedes/*microbiology ; Animals ; Dengue/*prevention & control ; Female ; Male ; *Models, Theoretical ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods ; *Sex Ratio ; Wolbachia/*pathogenicity ; }, abstract = {The introduction of endosymbiont Wolbachia into laboratory-reared mosquito populations, which are then released to mix with natural populations to prevent the mosquito vectors from reproducing and thus break the transmission cycle of dengue disease, is an innovative new technology. Field trials of Wolbachia-carrying mosquitoes have now been implemented in many countries where there have been the outbreaks of dengue disease. A mathematical model is proposed to investigate the effects of non-identical sex ratio releases of Wolbachia-carrying mosquitoes on the control of dengue transmission. Firstly, we analyzed the existence and stability of equilibria for the system and proved the existence of forward and backward bifurcations. Secondly, bifurcation diagrams, the basins of attraction of the equilibria and the effects of mosquito augmentation for the system with imperfect and perfect transmission rates were obtained. Thirdly, three possible results for mosquito augmentation were summarized for different parameter regions. Further we explored an uncertainty and sensitivity analysis of solutions to estimate the effects of different parameter values on the success or failure of population replacement. Based on the above analysis, we considered a series of relevant issues such as (a) whether or not mosquito augmentation can ensure the success of population replacement? (b) If not, what are the parameter regions for the success or possible success of population replacement? (c) How does the initial density of natural mosquitoes and the quantity of mosquito augmentations affect the success of population replacement? (d) Whether all population replacements are effective for reducing the spread of dengue virus in the end? The results of this study will be helpful for public health authorities in designing proper strategies of mosquito augmentations for the control of dengue disease.}, } @article {pmid29526588, year = {2018}, author = {Turelli, M and Cooper, BS and Richardson, KM and Ginsberg, PS and Peckenpaugh, B and Antelope, CX and Kim, KJ and May, MR and Abrieux, A and Wilson, DA and Bronski, MJ and Moore, BR and Gao, JJ and Eisen, MB and Chiu, JC and Conner, WR and Hoffmann, AA}, title = {Rapid Global Spread of wRi-like Wolbachia across Multiple Drosophila.}, journal = {Current biology : CB}, volume = {28}, number = {6}, pages = {963-971.e8}, pmid = {29526588}, issn = {1879-0445}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; R35 GM124701/GM/NIGMS NIH HHS/United States ; S10 RR027303/RR/NCRR NIH HHS/United States ; S10 RR029668/RR/NCRR NIH HHS/United States ; /HHMI/Howard Hughes Medical Institute/United States ; }, mesh = {Animals ; Biological Evolution ; DNA, Mitochondrial/analysis/genetics ; Disease Transmission, Infectious/veterinary ; Drosophila/*genetics/microbiology ; Evolution, Molecular ; Genome/genetics ; Infectious Disease Transmission, Vertical/veterinary ; Introduced Species ; Phylogeny ; Symbiosis/genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Maternally transmitted Wolbachia, Spiroplasma, and Cardinium bacteria are common in insects [1], but their interspecific spread is poorly understood. Endosymbionts can spread rapidly within host species by manipulating host reproduction, as typified by the global spread of wRi Wolbachia observed in Drosophila simulans [2, 3]. However, because Wolbachia cannot survive outside host cells, spread between distantly related host species requires horizontal transfers that are presumably rare [4-7]. Here, we document spread of wRi-like Wolbachia among eight highly diverged Drosophila hosts (10-50 million years) over only about 14,000 years (5,000-27,000). Comparing 110 wRi-like genomes, we find ≤0.02% divergence from the wRi variant that spread rapidly through California populations of D. simulans. The hosts include both globally invasive species (D. simulans, D. suzukii, and D. ananassae) and narrowly distributed Australian endemics (D. anomalata and D. pandora) [8]. Phylogenetic analyses that include mtDNA genomes indicate introgressive transfer of wRi-like Wolbachia between closely related species D. ananassae, D. anomalata, and D. pandora but no horizontal transmission within species. Our analyses suggest D. ananassae as the Wolbachia source for the recent wRi invasion of D. simulans and D. suzukii as the source of Wolbachia in its sister species D. subpulchrella. Although six of these wRi-like variants cause strong cytoplasmic incompatibility, two cause no detectable reproductive effects, indicating that pervasive mutualistic effects [9, 10] complement the reproductive manipulations for which Wolbachia are best known. "Super spreader" variants like wRi may be particularly useful for controlling insect pests and vector-borne diseases with Wolbachia transinfections [11].}, } @article {pmid29520067, year = {2018}, author = {Woodford, L and Bianco, G and Ivanova, Y and Dale, M and Elmer, K and Rae, F and Larcombe, SD and Helm, B and Ferguson, HM and Baldini, F}, title = {Vector species-specific association between natural Wolbachia infections and avian malaria in black fly populations.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {4188}, pmid = {29520067}, issn = {2045-2322}, mesh = {Animals ; Birds ; Haemosporida/*physiology ; *Insect Vectors/microbiology/parasitology ; *Malaria, Avian/epidemiology/microbiology/parasitology/transmission ; *Rickettsiaceae Infections/epidemiology/parasitology/transmission ; *Simuliidae/microbiology/parasitology ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Artificial infection of mosquitoes with the endosymbiont bacteria Wolbachia can interfere with malaria parasite development. Therefore, the release of Wolbachia-infected mosquitoes has been proposed as a malaria control strategy. However, Wolbachia effects on vector competence are only partly understood, as indicated by inconsistent effects on malaria infection reported under laboratory conditions. Studies of naturally-occurring Wolbachia infections in wild vector populations could be useful to identify the ecological and evolutionary conditions under which these endosymbionts can block malaria transmission. Here we demonstrate the occurrence of natural Wolbachia infections in three species of black fly (genus Simulium), which is a main vector of the avian malaria parasite Leucocytozoon. Prevalence of Leucocytozoon was high (25%), but the nature and magnitude of its association with Wolbachia differed between black fly species. Wolbachia infection was positively associated with avian malaria infection in S. cryophilum, negatively associated in S. aureum, and unrelated in S. vernum. These differences suggest that Wolbachia interacts with the parasite in a vector host species-specific manner. This provides a useful model system for further study of how Wolbachia influences vector competence. Such knowledge, including the possibility of undesirable positive association, is required to guide endosymbiont based control methods.}, } @article {pmid29519772, year = {2018}, author = {Wattanamethanont, J and Kaewthamasorn, M and Tiawsirisup, S}, title = {Natural infection of questing ixodid ticks with protozoa and bacteria in Chonburi Province, Thailand.}, journal = {Ticks and tick-borne diseases}, volume = {9}, number = {3}, pages = {749-758}, doi = {10.1016/j.ttbdis.2018.02.020}, pmid = {29519772}, issn = {1877-9603}, mesh = {Anaplasma/genetics/isolation & purification ; Animals ; Animals, Wild/microbiology/parasitology ; Babesia/genetics/isolation & purification ; Bacteria/genetics/*isolation & purification/pathogenicity ; DNA, Protozoan ; Humans ; Ixodidae/classification/genetics/*microbiology/*parasitology ; Larva/microbiology/parasitology ; Parasites/genetics/*isolation & purification ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Thailand/epidemiology ; Theileria/genetics/isolation & purification ; Tick-Borne Diseases/*epidemiology/microbiology/parasitology/transmission ; }, abstract = {Ixodid ticks are important vectors of tick-borne disease agents affecting humans and animals, with wildlife often serving as important reservoirs. This study examined protozoal and bacterial infection in questing ticks in forest habitats in Chonburi Province, Thailand in 2015, using PCR and DNA sequencing techniques. A total of 12,184 ticks were morphologically identified to species and a subset of ticks were confirmed by PCR, targeting the tick mitochondrial 16S rRNA gene. Tick species collected included Haemaphysalis lagrangei (92.8%), H. wellingtoni (0.1%), and Rhipicephalus microplus (7.0%). In total, 419 tick pools [ELM(1] [ST2] were examined by PCR amplification of a fragment of the 18S rRNA gene of Babesia and Theileria species, and the 16S rRNA gene of bacteria in the family Anaplasmataceae. Results revealed a tick infection rate for the tick pools of 57.0% (239/419) including four pathogens and one bacterial symbiont. The highest infection rate in H. lagrangei, H. wellingtoni, and R. microplus pools was recorded for Anaplasma spp. at 55.6% (233/419) including three Anaplasma species genotype groups Anaplasma spp. closely related to A. bovis, A. platys, and unidentified Anaplasma spp. Theileria spp. showed a lower infection rate in H. lagrangei at 4.3% (18/419) with three Theileria species genotypes closely related to T. cervi, T. capreoli, and unidentified Theileria spp. Only 0.2% (1/419) of H. lagrangei pools contained Babesia spp., Ehrlichia spp., or Wolbachia spp. [ELM(3] [ST4] These findings provided information on tick species in wildlife habitats and detected protozoa and bacteria in the ticks. The results suggest that these tick species are possible vectors for transmitting tick-borne disease agents in Thailand wildlife habitats.}, } @article {pmid29516174, year = {2018}, author = {Michalik, A and Szwedo, J and Stroiński, A and Świerczewski, D and Szklarzewicz, T}, title = {Symbiotic cornucopia of the monophagous planthopper Ommatidiotus dissimilis (Fallén, 1806) (Hemiptera: Fulgoromorpha: Caliscelidae).}, journal = {Protoplasma}, volume = {255}, number = {5}, pages = {1317-1329}, pmid = {29516174}, issn = {1615-6102}, support = {DS/MND/WBiNoZ/IZ/3/2015//Ministerstwo Nauki i Szkolnictwa Wyższego/ ; }, mesh = {Animals ; Enterobacteriaceae/genetics/physiology ; Hemiptera/*microbiology ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/physiology ; Symbiosis/genetics/physiology ; Wolbachia/genetics/physiology ; }, abstract = {In contrast to Cicadomorpha, in which numerous symbiotic bacteria have been identified and characterized, the symbionts of fulgoromorphans are poorly known. Here, we present the results of histological, ultrastructural, and molecular analyses of the symbiotic system of the planthopper Ommatidiotus dissimilis. Amplification, cloning, and sequencing of bacterial 16S RNA genes have revealed that O. dissimilis is host to five types of bacteria. Apart from bacteria Sulcia and Vidania, which are regarded as ancestral symbionts of Fulgoromorpha, three additional types of bacteria belonging to the genera Sodalis, Wolbachia, and Rickettsia have been detected. Histological and ultrastructural investigations have shown that bacteria Sulcia, Vidania, and Sodalis house separate bacteriocytes, whereas bacteria Wolbachia and Rickettsia are dispersed within various insect tissue. Additionally, bacteria belonging to the genus Vidania occupy the bacteriome localized in the lumen of the hindgut. Both molecular and microscopic analyses have revealed that all the symbionts are transovarially transmitted between generations.}, } @article {pmid29608200, year = {2017}, author = {Sseruwagi, P and Wainaina, J and Ndunguru, J and Tumuhimbise, R and Tairo, F and Guo, JY and Vrielink, A and Blythe, A and Kinene, T and De Marchi, B and Kehoe, MA and Tanz, S and Boykin, LM}, title = {The first transcriptomes from field-collected individual whiteflies (Bemisia tabaci, Hemiptera: Aleyrodidae): a case study of the endosymbiont composition.}, journal = {Gates open research}, volume = {1}, number = {}, pages = {16}, pmid = {29608200}, issn = {2572-4754}, abstract = {Background: Bemisia tabaci species (B. tabaci), or whiteflies, are the world's most devastating insect pests. They cause billions of dollars (US) of damage each year, and are leaving farmers in the developing world food insecure. Currently, all publically available transcriptome data for B. tabaci are generated from pooled samples, which can lead to high heterozygosity and skewed representation of the genetic diversity. The ability to extract enough RNA from a single whitefly has remained elusive due to their small size and technological limitations. Methods: In this study, we optimised a single whitefly RNA extraction procedure, and sequenced the transcriptome of four individual adult Sub-Saharan Africa 1 (SSA1) B. tabaci. Transcriptome sequencing resulted in 39-42 million raw reads. De novo assembly of trimmed reads yielded between 65,000-162,000 Contigs across B. tabaci transcriptomes. Results: Bayesian phylogenetic analysis of mitochondrion cytochrome I oxidase (mtCOI) grouped the four whiteflies within the SSA1 clade. BLASTn searches on the four transcriptomes identified five endosymbionts; the primary endosymbiont Portiera aleyrodidarum and four secondary endosymbionts: Arsenophonus, Wolbachia, Rickettsia, and Cardinium spp. that were predominant across all four SSA1 B. tabaci samples with prevalence levels of between 54.1 to 75%. Amino acid alignments of the NusG gene of P. aleyrodidarum for the SSA1 B. tabaci transcriptomes of samples WF2 and WF2b revealed an eleven amino acid residue deletion that was absent in samples WF1 and WF2a. Comparison of the protein structure of the NusG protein from P. aleyrodidarum in SSA1 with known NusG structures showed the deletion resulted in a shorter D loop. Conclusions: The use of field-collected specimens means time and money will be saved in future studies using single whitefly transcriptomes in monitoring vector and viral interactions. Our method is applicable to any small organism where RNA quantity has limited transcriptome studies.}, } @article {pmid29499213, year = {2018}, author = {Bi, J and Sehgal, A and Williams, JA and Wang, YF}, title = {Wolbachia affects sleep behavior in Drosophila melanogaster.}, journal = {Journal of insect physiology}, volume = {107}, number = {}, pages = {81-88}, doi = {10.1016/j.jinsphys.2018.02.011}, pmid = {29499213}, issn = {1879-1611}, mesh = {Animals ; Arousal ; Circadian Rhythm ; Drosophila melanogaster/*microbiology/*physiology ; Female ; Insect Proteins ; Male ; *Sleep ; Symbiosis ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria present in a wide range of insects. Although their dramatic effects on host reproductive biology have been well studied, the effects of Wolbachia on sleep behavior of insect hosts are not well documented. In this study, we report that Wolbachia infection caused an increase of total sleep time in both male and female Drosophila melanogaster. The increase in sleep was associated with an increase in the number of nighttime sleep bouts or episodes, but not in sleep bout duration. Correspondingly, Wolbachia infection also reduced the arousal threshold of their fly hosts. However, neither circadian rhythm nor sleep rebound following deprivation was influenced by Wolbachia infection. Transcriptional analysis of the dopamine biosynthesis pathway revealed that two essential genes, Pale and Ddc, were significantly upregulated in Wolbachia-infected flies. Together, these results indicate that Wolbachia mediates the expression of dopamine related genes, and decreases the sleep quality of their insect hosts. Our findings help better understand the host-endosymbiont interactions and in particular the Wolbachia's impact on behaviors, and thus on ecology and evolution in insect hosts.}, } @article {pmid29497475, year = {2018}, author = {Hashmi, TR and Devi, SR and Meshram, NM and Prasad, R}, title = {Assessment of bacterial endosymbionts and the host, Bemisia tabaci (Hemiptera: Aleyrodidae), using rRNA and mitochondrial cytochrome oxidase I gene sequences.}, journal = {Communicative & integrative biology}, volume = {11}, number = {1}, pages = {e1433442}, pmid = {29497475}, issn = {1942-0889}, abstract = {Endosymbionts are vital factor for arthropod ecology. The whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a cryptic species complex composed of more than 34 putative species. Moreover to the primary endosymbiont Portiera aleyrodidarum, six secondary endosymbionts Cardinium, Arsenophonus, Rickettsia, Wolbachia, Hamiltonella and Fritschea are known in B. tabaci. Here, we tested four of the six secondary endosymbiont lineages (excluding Fritschea and Hamiltonella) from 180 whitely individuals collected from six host plants belonging to families Solanaceae (Brinjal, Tomato and Potato) and Fabaceae (Soyabean, Mungbean and Subabool). Phylogenetic studies grounded on the mitochondrial cytochrome I gene revealed the presence of Asia 1, Asia II 1 and Asia II 7 genetic groups for B. tabaci. Specific primers targeting 16S rRNA and 23S rRNA gene were used for estimating the bacterial endosymbionts. As a primary endosymbiont Portiera aleyrodidarum was present in all the studied samples; whereas, an uneven distribution of secondary endosymbionts were recorded. Overall our finding exposes the variation and diversity of endosymbionts within the B. tabaci collected from different host plants and outlines the genetic groups of the insect pest. The study delivers a significant information concerning the circulation of secondary endosymbionts with host preferences of B. tabaci and provides suggestion for progressive studies on targeting the specific endosymbionts with respect to host for the control measures.}, } @article {pmid29494679, year = {2018}, author = {Thomas, S and Verma, J and Woolfit, M and O'Neill, SL}, title = {Wolbachia-mediated virus blocking in mosquito cells is dependent on XRN1-mediated viral RNA degradation and influenced by viral replication rate.}, journal = {PLoS pathogens}, volume = {14}, number = {3}, pages = {e1006879}, pmid = {29494679}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*immunology/metabolism/microbiology/virology ; Animals ; Dengue/immunology/*prevention & control/virology ; Dengue Virus/physiology ; Exoribonucleases/genetics/*metabolism ; Humans ; Insect Vectors/immunology/microbiology/virology ; MicroRNAs ; Microtubule-Associated Proteins/genetics/*metabolism ; RNA, Viral/genetics/*metabolism ; Symbiosis ; *Virus Replication ; West Nile Fever/immunology/*prevention & control/virology ; West Nile virus/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is currently being developed as a novel tool to block the transmission of dengue viruses (DENV) by Aedes aegypti. A number of mechanisms have been proposed to explain the DENV-blocking phenotype in mosquitoes, including competition for fatty acids like cholesterol, manipulation of host miRNAs and upregulation of innate immune pathways in the mosquito. We examined the various stages in the DENV infection process to better understand the mechanism of Wolbachia-mediated virus blocking (WMVB). Our results suggest that infection with Wolbachia does not inhibit DENV binding or cell entry, but reduces virus replication. In contrast to a previous report, we also observed a similar reduction in replication of West Nile virus (WNV). This reduced replication is associated with rapid viral RNA degradation in the cytoplasm. We didn't find a role for host miRNAs in WMVB. Further analysis showed that the 3' end of the virus subgenomic RNA was protected and accumulated over time suggesting that the degradation is XRN1-mediated. We also found that sub genomic flavivirus RNA accumulation inactivated XRN1 in mosquito cells in the absence of Wolbachia and led to enhancement of RNA degradation in its presence. Depletion of XRN1 decreased WMVB which was associated with a significant increase in DENV RNA. We also observed that WMVB is influenced by virus MOI and rate of virus replication. A comparatively elevated blocking was observed for slowly replicating DENV, compared to WNV. Similar results were obtained while analysing different DENV serotypes.}, } @article {pmid29487575, year = {2018}, author = {Aydogan, EL and Moser, G and Müller, C and Kämpfer, P and Glaeser, SP}, title = {Long-Term Warming Shifts the Composition of Bacterial Communities in the Phyllosphere of Galium album in a Permanent Grassland Field-Experiment.}, journal = {Frontiers in microbiology}, volume = {9}, number = {}, pages = {144}, pmid = {29487575}, issn = {1664-302X}, abstract = {Global warming is currently a much discussed topic with as yet largely unexplored consequences for agro-ecosystems. Little is known about the warming effect on the bacterial microbiota inhabiting the plant surface (phyllosphere), which can have a strong impact on plant growth and health, as well as on plant diseases and colonization by human pathogens. The aim of this study was to investigate the effect of moderate surface warming on the diversity and composition of the bacterial leaf microbiota of the herbaceous plant Galium album. Leaves were collected from four control and four surface warmed (+2°C) plots located at the field site of the Environmental Monitoring and Climate Impact Research Station Linden in Germany over a 6-year period. Warming had no effect on the concentration of total number of cells attached to the leaf surface as counted by Sybr Green I staining after detachment, but changes in the diversity and phylogenetic composition of the bacterial leaf microbiota analyzed by bacterial 16S rRNA gene Illumina amplicon sequencing were observed. The bacterial phyllosphere microbiota were dominated by Proteobacteria, Bacteroidetes, and Actinobacteria. Warming caused a significant higher relative abundance of members of the Gammaproteobacteria, Actinobacteria, and Firmicutes, and a lower relative abundance of members of the Alphaproteobacteria and Bacteroidetes. Plant beneficial bacteria like Sphingomonas spp. and Rhizobium spp. occurred in significantly lower relative abundance in leaf samples of warmed plots. In contrast, several members of the Enterobacteriaceae, especially Enterobacter and Erwinia, and other potential plant or human pathogenic genera such as Acinetobacter and insect-associated Buchnera and Wolbachia spp. occurred in higher relative abundances in the phyllosphere samples from warmed plots. This study showed for the first time the long-term impact of moderate (+2°C) surface warming on the phyllosphere microbiota on plants. A reduction of beneficial bacteria and an enhancement of potential pathogenic bacteria in the phyllosphere of plants may indicate that this aspect of the ecosystem which has been largely neglected up till now, can be a potential risk for pathogen transmission in agro-ecosystems in the near future.}, } @article {pmid29471864, year = {2018}, author = {Gesto, JSM and Araki, AS and Caragata, EP and de Oliveira, CD and Martins, AJ and Bruno, RV and Moreira, LA}, title = {In tune with nature: Wolbachia does not prevent pre-copula acoustic communication in Aedes aegypti.}, journal = {Parasites & vectors}, volume = {11}, number = {1}, pages = {109}, pmid = {29471864}, issn = {1756-3305}, mesh = {Aedes/microbiology/*physiology ; *Animal Communication ; Animals ; Female ; Genetic Fitness ; Male ; Mosquito Control ; Mosquito Vectors/microbiology/*physiology ; *Sexual Behavior, Animal ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Mosquito-borne diseases are rapidly spreading to vast territories, putting at risk most of the world's population. A key player in this scenario is Aedes aegypti, a hematophagous species which hosts and transmits viruses causing dengue and other serious illnesses. Since vector control strategies relying only on insecticides have proven unsustainable, an alternative method involving the release of Wolbachia-harboring individuals has emerged. Its successful implementation vastly depends on how fit the released individuals are in the natural habitat, being able to mate with wild populations and to spread Wolbachia to subsequent generations. In mosquitoes, an important aspect of reproductive fitness is the acoustic communication between males and females, which translates to interactions between harmonic frequencies in close proximity flight. This study aimed to characterize the flight tone produced by individuals harboring Wolbachia, also evaluating their ability to establish stable acoustic interactions.

METHODS: Wild-type (WT) and Wolbachia-harboring specimens (wMelBr) were thorax-tethered to blunt copper wires and placed at close proximity to sensitive microphones. Wing-beat frequencies (WBFs) were characterized at fundamental and harmonic levels, for both single individuals and couples. Harmonic interactions in homogeneous and heterogeneous couples of WT and wMelBr variants were identified, categorized and quantified accordingly.

RESULTS: In tethered 'solo' flights, individuals harboring Wolbachia developed WBFs, differing slightly, in a sex-dependent way, from those of the WT strain. To test the ability to form harmonic 'duets', tethered couples of wMelBr and WT individuals were shuffled in different sex pairs and had their flight tones analyzed. All couple types, with WT and/or wMelBr individuals, were able to interact acoustically in the frequency range of 1300-1500 Hz, which translates to the convergence between male's second harmonic and female's third. No significant differences were found in the proportions of interacting couples between the pair types. Surprisingly, spectrograms also revealed the convergence between alternative harmonic frequencies, inside and outside the species putative hearing threshold.

CONCLUSIONS: Wolbachia infection leads to small sex-dependent changes on the flight tones of Ae. aegypti, but it does not seem to prevent the stereotyped harmonic interaction between males and females. Therefore, when released in the natural habitat to breed with native individuals, Wolbachia-harboring individuals shall be fit enough to meet the criteria of acoustically-related mating behavior and promote bacteria dispersion effectively.}, } @article {pmid29468305, year = {2018}, author = {Ramalho, MO and Vieira, AS and Pereira, MC and Moreau, CS and Bueno, OC}, title = {Transovarian Transmission of Blochmannia and Wolbachia Endosymbionts in the Neotropical Weaver Ant Camponotus textor (Hymenoptera, Formicidae).}, journal = {Current microbiology}, volume = {75}, number = {7}, pages = {866-873}, pmid = {29468305}, issn = {1432-0991}, support = {007343/2014-00//CAPES Foundation/ ; 157837/2015-7//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; }, mesh = {Animals ; Ants/growth & development/*microbiology ; Enterobacteriaceae/classification/genetics/*isolation & purification/physiology ; Female ; In Situ Hybridization, Fluorescence ; Ovary/growth & development/microbiology ; Ovum/growth & development/microbiology ; *Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Camponotus is a hyper-diverse ant genus that is associated with the obligate endosymbiont Blochmannia, and often also with Wolbachia, but morphological studies on the location of these bacteria in the queen's ovaries during oogenesis remain limited. In the present study, we used the Neotropical weaver ant Camponotus textor to characterize the ovary using histology (HE) techniques, and to document the location of Blochmannia and Wolbachia during oogenesis through fluorescence in situ hybridization (FISH). This is the first morphological report of these two bacteria in the same host with polytrophic meroistic ovaries and reveals that Blochmannia is found inside late-stage oocytes and Wolbachia is associated with the nuclei of the nurse cells. Our results provide insights into the developmental sequence of when these bacteria reach the egg, with Blochmannia establishing itself in the egg first, and Wolbachia only reaching the egg shortly before completing egg development. Studies such as this provide understanding about the mechanisms and timing of the establishment of these endosymbionts in the host.}, } @article {pmid29468014, year = {2018}, author = {Zu Dohna, H and Houry, C and Kambris, Z}, title = {A comparative analysis of Wolbachia-induced host reproductive phenotypes reveals transition rate heterogeneity.}, journal = {Ecology and evolution}, volume = {8}, number = {4}, pages = {1945-1953}, pmid = {29468014}, issn = {2045-7758}, abstract = {The endosymbiotic bacterium Wolbachia infects a wide range of arthropods and their relatives. It is an intracellular parasite transmitted through the egg from mother to offspring. Wolbachia can spread and persist through various means of host reproductive manipulation. How these different mechanisms of host manipulation evolved in Wolbachia is unclear. Which host reproductive phenotype is most likely to be ancestral and whether evolutionary transitions between some host phenotypes are more common than others remain unanswered questions. Recent studies have revealed multiple cases where the same Wolbachia strain can induce different reproductive phenotypes in different hosts, raising the question to what degree the induced host phenotype should be regarded as a trait of Wolbachia. In this study, we constructed a phylogenetic tree of Wolbachia and analyzed the patterns of host phenotypes along that tree. We were able to detect a phylogenetic signal of host phenotypes on the Wolbachia tree, indicating that the induced host phenotype can be regarded as a Wolbachia trait. However, we found no clear support for the previously stated hypothesis that cytoplasmic incompatibility is ancestral to Wolbachia in arthropods. Our analysis provides evidence for heterogeneous transition rates between host phenotypes.}, } @article {pmid29467241, year = {2018}, author = {Kamath, AD and Deehan, MA and Frydman, HM}, title = {Polar cell fate stimulates Wolbachia intracellular growth.}, journal = {Development (Cambridge, England)}, volume = {145}, number = {6}, pages = {}, pmid = {29467241}, issn = {1477-9129}, support = {R56 AI097589/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*microbiology ; Female ; Fluorescent Antibody Technique ; In Situ Hybridization, Fluorescence ; Oogenesis/physiology ; Ovary/cytology/microbiology ; Polar Bodies/*microbiology ; Symbiosis/*physiology ; Viral Tropism/*physiology ; Wolbachia/*pathogenicity ; }, abstract = {Bacteria are crucial partners in the development and evolution of vertebrates and invertebrates. A large fraction of insects harbor Wolbachia, bacterial endosymbionts that manipulate host reproduction to favor their spreading. Because they are maternally inherited, Wolbachia are under selective pressure to reach the female germline and infect the offspring. However, Wolbachia infection is not limited to the germline. Somatic cell types, including stem cell niches, have higher Wolbachia loads compared with the surrounding tissue. Here, we show a novel Wolbachia tropism to polar cells (PCs), specialized somatic cells in the Drosophila ovary. During oogenesis, all stages of PC development are easily visualized, facilitating the investigation of the kinetics of Wolbachia intracellular growth. Wolbachia accumulation is triggered by particular events of PC morphogenesis, including differentiation from progenitors and between stages 8 and 9 of oogenesis. Moreover, induction of ectopic PC fate is sufficient to promote Wolbachia accumulation. We found that Wolbachia PC tropism is evolutionarily conserved across most Drosophila species, but not in Culex mosquitos. These findings highlight the coordination of endosymbiont tropism with host development and cell differentiation.}, } @article {pmid29466376, year = {2018}, author = {Yadav, S and Frazer, J and Banga, A and Pruitt, K and Harsh, S and Jaenike, J and Eleftherianos, I}, title = {Endosymbiont-based immunity in Drosophila melanogaster against parasitic nematode infection.}, journal = {PloS one}, volume = {13}, number = {2}, pages = {e0192183}, pmid = {29466376}, issn = {1932-6203}, mesh = {Animals ; Drosophila melanogaster/*immunology/microbiology/parasitology ; Host-Parasite Interactions ; Nematoda/*physiology ; *Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {Associations between endosymbiotic bacteria and their hosts represent a complex ecosystem within organisms ranging from humans to protozoa. Drosophila species are known to naturally harbor Wolbachia and Spiroplasma endosymbionts, which play a protective role against certain microbial infections. Here, we investigated whether the presence or absence of endosymbionts affects the immune response of Drosophila melanogaster larvae to infection by Steinernema carpocapsae nematodes carrying or lacking their mutualistic Gram-negative bacteria Xenorhabdus nematophila (symbiotic or axenic nematodes, respectively). We find that the presence of Wolbachia alone or together with Spiroplasma promotes the survival of larvae in response to infection with S. carpocapsae symbiotic nematodes, but not against axenic nematodes. We also find that Wolbachia numbers are reduced in Spiroplasma-free larvae infected with axenic compared to symbiotic nematodes, and they are also reduced in Spiroplasma-containing compared to Spiroplasma-free larvae infected with axenic nematodes. We further show that S. carpocapsae axenic nematode infection induces the Toll pathway in the absence of Wolbachia, and that symbiotic nematode infection leads to increased phenoloxidase activity in D. melanogaster larvae devoid of endosymbionts. Finally, infection with either type of nematode alters the metabolic status and the fat body lipid droplet size in D. melanogaster larvae containing only Wolbachia or both endosymbionts. Our results suggest an interaction between Wolbachia endosymbionts with the immune response of D. melanogaster against infection with the entomopathogenic nematodes S. carpocapsae. Results from this study indicate a complex interplay between insect hosts, endosymbiotic microbes and pathogenic organisms.}, } @article {pmid29447357, year = {2019}, author = {Jewell, NP and Dufault, S and Cutcher, Z and Simmons, CP and Anders, KL}, title = {Analysis of cluster-randomized test-negative designs: cluster-level methods.}, journal = {Biostatistics (Oxford, England)}, volume = {20}, number = {2}, pages = {332-346}, pmid = {29447357}, issn = {1468-4357}, support = {R56 AI134724/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/microbiology ; Animals ; Biostatistics/*methods ; Dengue/prevention & control ; Humans ; *Models, Statistical ; Mosquito Vectors ; *Random Allocation ; Randomized Controlled Trials as Topic/*methods ; Wolbachia/pathogenicity ; }, abstract = {Intervention trials of vector control methods often require community level randomization with appropriate inferential methods. For many interventions, the possibility of confounding due to the effects of health-care seeking behavior on disease ascertainment remains a concern. The test-negative design, a variant of the case-control method, was introduced to mitigate this issue in the assessment of the efficacy of influenza vaccination (measured at an individual level) on influenza infection. Here, we introduce a cluster-randomized test-negative design that includes randomization of the intervention at a group level. We propose several methods for estimation and inference regarding the relative risk (RR). The inferential methods considered are based on the randomization distribution induced by permuting intervention assignment across two sets of randomly selected clusters. The motivating example is a current study of the efficacy of randomized releases of Wolbachia-infected Aedes aegypti mosquitoes to reduce the incidence of dengue in Yogyakarta City, Indonesia. Estimation and inference techniques are assessed through a simulation study.}, } @article {pmid29443908, year = {2018}, author = {Lim, EXY and Lee, WS and Madzokere, ET and Herrero, LJ}, title = {Mosquitoes as Suitable Vectors for Alphaviruses.}, journal = {Viruses}, volume = {10}, number = {2}, pages = {}, pmid = {29443908}, issn = {1999-4915}, mesh = {Alphavirus/*physiology ; Alphavirus Infections/*transmission/*virology ; Animals ; Coinfection ; Culicidae/*virology ; Humans ; Mosquito Vectors/*virology ; Mutation ; RNA, Viral ; }, abstract = {Alphaviruses are arthropod-borne viruses and are predominantly transmitted via mosquito vectors. This vector preference by alphaviruses raises the important question of the determinants that contribute to vector competence. There are several tissue barriers of the mosquito that the virus must overcome in order to establish a productive infection. Of importance are the midgut, basal lamina and the salivary glands. Infection of the salivary glands is crucial for virus transmission during the mosquito's subsequent bloodfeed. Other factors that may contribute to vector competence include the microflora and parasites present in the mosquito, environmental conditions, the molecular determinants of the virus to adapt to the vector, as well as the effect of co-infection with other viruses. Though mosquito innate immunity is a contributing factor to vector competence, it will not be discussed in this review. Detailed understanding of these factors will be instrumental in minimising transmission of alphaviral diseases.}, } @article {pmid29435771, year = {2018}, author = {Zélé, F and Weill, M and Magalhães, S}, title = {Identification of spider-mite species and their endosymbionts using multiplex PCR.}, journal = {Experimental & applied acarology}, volume = {74}, number = {2}, pages = {123-138}, pmid = {29435771}, issn = {1572-9702}, support = {FCT-ANR//BIA-EVF/0013/2012//Faculdade de Ciências e Tecnologia (FCT), Universidade de Lisboa (PT)/ ; FCT-TUBITAK/0001/2014//Faculdade de Ciências e Tecnologia (FCT), Universidade de Lisboa (PT)/ ; SFRH/BPD/125020/2016//Faculdade de Ciências e Tecnologia (FCT), Universidade de Lisboa (PT)/ ; }, mesh = {Animals ; Bacteria/*isolation & purification ; Bacterial Physiological Phenomena ; Female ; Male ; *Microbiota ; Multiplex Polymerase Chain Reaction/economics/*methods ; Portugal ; Spain ; *Symbiosis ; Tetranychidae/*classification/genetics/microbiology ; }, abstract = {Spider mites of the genus Tetranychidae are severe crop pests. In the Mediterranean a few species coexist, but they are difficult to identify based on morphological characters. Additionally, spider mites often harbour several species of endosymbiotic bacteria, which may affect the biology of their hosts. Here, we propose novel, cost-effective, multiplex diagnostic methods allowing a quick identification of spider-mite species as well as of the endosymbionts they carry. First, we developed, and successfully multiplexed in a single PCR, primers to identify Tetranychus urticae, T. evansi and T. ludeni, some of the most common tetranychids found in southwest Europe. Moreover, we demonstrated that this method allows detecting multiple species in a single pool, even at low frequencies (up to 1/100), and can be used on entire mites without DNA extraction. Second, we developed another set of primers to detect spider-mite endosymbionts, namely Wolbachia, Cardinium and Rickettsia in a multiplex PCR, along with a generalist spider-mite primer to control for potential failure of DNA amplification in each PCR. Overall, our method represents a simple, cost-effective and reliable method to identify spider-mite species and their symbionts in natural field populations, as well as to detect contaminations in laboratory rearings. This method may easily be extended to other species.}, } @article {pmid29435238, year = {2018}, author = {Zhang, YK and Yang, K and Zhu, YX and Hong, XY}, title = {Symbiont-conferred reproduction and fitness benefits can favour their host occurrence.}, journal = {Ecology and evolution}, volume = {8}, number = {3}, pages = {1626-1633}, pmid = {29435238}, issn = {2045-7758}, abstract = {Double infections of Wolbachia and Spiroplasma are frequent in natural populations of Tetranychus truncatus, a polyphagous mite species that has been a dominant species in China since 2009. However, little is known about the causes and ecological importance of such coexistences. In this study, we established T. truncatus strains with different infection types and then inferred the impact of the two endosymbionts on host reproduction and fitness. Double infection induced cytoplasmic incompatibility, which was demonstrated by reduction in egg hatchability of incompatible crosses. However, doubly infected females produced more eggs relative to other strains. Wolbachia and Spiroplasma did not affect host survival, whereas doubly infected females and males developed faster than other strains. Such reproduction and fitness benefits provided by double infections may be associated with the lower densities of each symbiont, and the quantitative results also confirmed competition between Wolbachia and Spiroplasma in doubly infected females. These symbiont-conferred beneficial effects maintain stable prevalence of the symbionts and also help drive T. truncatus outbreaks in combination with other environmental factors.}, } @article {pmid29429122, year = {2018}, author = {Campo-Duarte, DE and Vasilieva, O and Cardona-Salgado, D and Svinin, M}, title = {Optimal control approach for establishing wMelPop Wolbachia infection among wild Aedes aegypti populations.}, journal = {Journal of mathematical biology}, volume = {76}, number = {7}, pages = {1907-1950}, pmid = {29429122}, issn = {1432-1416}, mesh = {Aedes/*microbiology/pathogenicity ; Animals ; Computational Biology ; Computer Simulation ; Dengue/prevention & control/transmission ; Female ; Host Microbial Interactions/physiology ; Humans ; Male ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/*microbiology/pathogenicity ; Pest Control, Biological/*methods/statistics & numerical data ; Population Dynamics/statistics & numerical data ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Wolbachia-based biocontrol has recently emerged as a potential method for prevention and control of dengue and other vector-borne diseases. Major vector species, such as Aedes aegypti females, when deliberately infected with Wolbachia become less capable of getting viral infections and transmitting the virus to human hosts. In this paper, we propose an explicit sex-structured population model that describes an interaction of uninfected (wild) male and female mosquitoes and those deliberately infected with wMelPop strain of Wolbachia in the same locality. This particular strain of Wolbachia is regarded as the best blocker of dengue and other arboviral infections. However, wMelPop strain of Wolbachia also causes the loss of individual fitness in Aedes aegypti mosquitoes. Our model allows for natural introduction of the decision (or control) variable, and we apply the optimal control approach to simulate wMelPop Wolbachia infestation of wild Aedes aegypti populations. The control action consists in continuous periodic releases of mosquitoes previously infected with wMelPop strain of Wolbachia in laboratory conditions. The ultimate purpose of control is to find a tradeoff between reaching the population replacement in minimum time and with minimum cost of the control effort. This approach also allows us to estimate the number of Wolbachia-carrying mosquitoes to be released in day-by-day control action. The proposed method of biological control is safe to human health, does not contaminate the environment, does not make harm to non-target species, and preserves their interaction with mosquitoes in the ecosystem.}, } @article {pmid29428509, year = {2018}, author = {Sahoo, RK and Lohman, DJ and Wahlberg, N and Müller, CJ and Brattström, O and Collins, SC and Peggie, D and Aduse-Poku, K and Kodandaramaiah, U}, title = {Evolution of Hypolimnas butterflies (Nymphalidae): Out-of-Africa origin and Wolbachia-mediated introgression.}, journal = {Molecular phylogenetics and evolution}, volume = {123}, number = {}, pages = {50-58}, doi = {10.1016/j.ympev.2018.02.001}, pmid = {29428509}, issn = {1095-9513}, mesh = {Africa ; Animals ; Base Sequence ; Bayes Theorem ; Biodiversity ; *Biological Evolution ; Butterflies/*genetics/*microbiology ; DNA, Mitochondrial/genetics ; Haplotypes/genetics ; Larva/physiology ; Likelihood Functions ; Mitochondria/genetics ; Phylogeny ; Phylogeography ; Sequence Analysis, DNA ; Wolbachia/*physiology ; }, abstract = {Hypolimnas butterflies (Nymphalidae), commonly known as eggflies, are a popular model system for studying a wide range of ecological questions including mimicry, polymorphism, wing pattern evolution, and Wolbachia-host interactions. The lack of a time-calibrated phylogeny for this group has precluded understanding its evolutionary history. We reconstruct a species-level phylogeny using a nine gene dataset and estimate species divergence times. Based on the resulting tree, we investigate the taxon's historical biogeography, examine the evolution of host plant preferences, and test the hypothesis that the endosymbiotic bacterium Wolbachia mediates gene transfer between species. Our analyses indicate that the species are grouped within three strongly supported, deeply divergent clades. However, relationships among these three clades are uncertain. In addition, many Hypolimnas species are not monophyletic or monophyletic with weak support, suggesting widespread incomplete lineage sorting and/or introgression. Biogeographic analysis strongly indicates that the genus diverged from its ancestor in Africa and subsequently dispersed to Asia; the strength of this result is not affected by topological uncertainties. While the larvae of African species feed almost exclusively on Urticaceae, larvae of species found further east often feed on several additional families. Interestingly, we found an identical mitochondrial haplotype in two Hypolimnas species, H. bolina and H. alimena, and a strong association between this mitotype and the Wolbachia strain wBol1a. Future investigations should explore the plausibility of Wolbachia-mediated introgression between species.}, } @article {pmid29396201, year = {2018}, author = {Ritchie, SA and van den Hurk, AF and Smout, MJ and Staunton, KM and Hoffmann, AA}, title = {Mission Accomplished? We Need a Guide to the 'Post Release' World of Wolbachia for Aedes-borne Disease Control.}, journal = {Trends in parasitology}, volume = {34}, number = {3}, pages = {217-226}, doi = {10.1016/j.pt.2017.11.011}, pmid = {29396201}, issn = {1471-5007}, mesh = {Aedes/*microbiology/*virology ; Animals ; Mosquito Vectors/*microbiology/*virology ; Pest Control, Biological/standards ; RNA Viruses/*physiology ; Wolbachia/*physiology ; }, abstract = {Historically, sustained control of Aedes aegypti, the vector of dengue, chikungunya, yellow fever, and Zika viruses, has been largely ineffective. Subsequently, two novel 'rear and release' control strategies utilizing mosquitoes infected with Wolbachia are currently being developed and deployed widely. In the incompatible insect technique, male Aedes mosquitoes, infected with Wolbachia, suppress populations through unproductive mating. In the transinfection strategy, both male and female Wolbachia-infected Ae. aegypti mosquitoes rapidly infect the wild population with Wolbachia, blocking virus transmission. It is critical to monitor the long-term stability of Wolbachia in host populations, and also the ability of this bacterium to continually inhibit virus transmission. Ongoing release and monitoring programs must be future-proofed should political support weaken when these vectors are successfully controlled.}, } @article {pmid29390142, year = {2018}, author = {Zélé, F and Santos, I and Olivieri, I and Weill, M and Duron, O and Magalhães, S}, title = {Endosymbiont diversity and prevalence in herbivorous spider mite populations in South-Western Europe.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {4}, pages = {}, doi = {10.1093/femsec/fiy015}, pmid = {29390142}, issn = {1574-6941}, mesh = {Animals ; Bacteroidetes/classification/genetics/*isolation & purification ; Biological Evolution ; Europe ; Herbivory ; Microbiota/genetics ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Rickettsia/classification/genetics/*isolation & purification ; Symbiosis ; Tetranychidae/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Bacterial endosymbionts are known as important players of the evolutionary ecology of their hosts. However, their distribution, prevalence and diversity are still largely unexplored. To this aim, we investigated infections by the most common bacterial reproductive manipulators in herbivorous spider mites of South-Western Europe. Across 16 populations belonging to three Tetranychus species, Wolbachia was the most prevalent (ca. 61%), followed by Cardinium (12%-15%), while only few individuals were infected by Rickettsia (0.9%-3%), and none carried Arsenophonus or Spiroplasma. These endosymbionts are here reported for the first time in Tetranychus evansi and Tetranychus ludeni, and showed variable infection frequencies between and within species, with several cases of coinfections. Moreover, Cardinium was more prevalent in Wolbachia-infected individuals, which suggests facilitation between these symbionts. Finally, sequence comparisons revealed no variation of the Wolbachia wsp and Rickettsia gtlA genes, but some diversity of the Cardinium 16S rRNA, both between and within populations of the three mite species. Some of the Cardinium sequences identified belonged to distantly-related clades, and the lack of association between these sequences and spider mite mitotypes suggests repeated host switching of Cardinium. Overall, our results reveal a complex community of symbionts in this system, opening the path for future studies.}, } @article {pmid29375803, year = {2018}, author = {Thongsripong, P and Chandler, JA and Green, AB and Kittayapong, P and Wilcox, BA and Kapan, DD and Bennett, SN}, title = {Mosquito vector-associated microbiota: Metabarcoding bacteria and eukaryotic symbionts across habitat types in Thailand endemic for dengue and other arthropod-borne diseases.}, journal = {Ecology and evolution}, volume = {8}, number = {2}, pages = {1352-1368}, pmid = {29375803}, issn = {2045-7758}, support = {P20 RR018727/RR/NCRR NIH HHS/United States ; U54 AI065359/AI/NIAID NIH HHS/United States ; }, abstract = {Vector-borne diseases are a major health burden, yet factors affecting their spread are only partially understood. For example, microbial symbionts can impact mosquito reproduction, survival, and vectorial capacity, and hence affect disease transmission. Nonetheless, current knowledge of mosquito-associated microbial communities is limited. To characterize the bacterial and eukaryotic microbial communities of multiple vector species collected from different habitat types in disease endemic areas, we employed next-generation 454 pyrosequencing of 16S and 18S rRNA amplicon libraries, also known as metabarcoding. We investigated pooled whole adult mosquitoes of three medically important vectors, Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus, collected from different habitats across central Thailand where we previously characterized mosquito diversity. Our results indicate that diversity within the mosquito microbiota is low, with the majority of microbes assigned to one or a few taxa. Two of the most common eukaryotic and bacterial genera recovered (Ascogregarina and Wolbachia, respectively) are known mosquito endosymbionts with potentially parasitic and long evolutionary relationships with their hosts. Patterns of microbial composition and diversity appeared to differ by both vector species and habitat for a given species, although high variability between samples suggests a strong stochastic element to microbiota assembly. In general, our findings suggest that multiple factors, such as habitat condition and mosquito species identity, may influence overall microbial community composition, and thus provide a basis for further investigations into the interactions between vectors, their microbial communities, and human-impacted landscapes that may ultimately affect vector-borne disease risk.}, } @article {pmid29370307, year = {2018}, author = {Ant, TH and Herd, CS and Geoghegan, V and Hoffmann, AA and Sinkins, SP}, title = {The Wolbachia strain wAu provides highly efficient virus transmission blocking in Aedes aegypti.}, journal = {PLoS pathogens}, volume = {14}, number = {1}, pages = {e1006815}, pmid = {29370307}, issn = {1553-7374}, support = {MC_PC_15087/MRC_/Medical Research Council/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; EP-D-16-021/EPA/EPA/United States ; }, mesh = {Aedes/*microbiology/*virology ; Alphavirus Infections/microbiology/transmission/virology ; Animals ; Antibiosis/*physiology ; *Dengue Virus/pathogenicity ; Female ; Infectious Disease Transmission, Vertical/*prevention & control/veterinary ; Inheritance Patterns ; Male ; Pest Control, Biological ; Semliki forest virus/pathogenicity/physiology ; Viral Load ; Wolbachia/*physiology ; }, abstract = {Introduced transinfections of the inherited bacteria Wolbachia can inhibit transmission of viruses by Aedes mosquitoes, and in Ae. aegypti are now being deployed for dengue control in a number of countries. Only three Wolbachia strains from the large number that exist in nature have to date been introduced and characterized in this species. Here novel Ae. aegypti transinfections were generated using the wAlbA and wAu strains. In its native Ae. albopictus, wAlbA is maintained at lower density than the co-infecting wAlbB, but following transfer to Ae. aegypti the relative strain density was reversed, illustrating the strain-specific nature of Wolbachia-host co-adaptation in determining density. The wAu strain also reached high densities in Ae. aegypti, and provided highly efficient transmission blocking of dengue and Zika viruses. Both wAu and wAlbA were less susceptible than wMel to density reduction/incomplete maternal transmission resulting from elevated larval rearing temperatures. Although wAu does not induce cytoplasmic incompatibility (CI), it was stably combined with a CI-inducing strain as a superinfection, and this would facilitate its spread into wild populations. Wolbachia wAu provides a very promising new option for arbovirus control, particularly for deployment in hot tropical climates.}, } @article {pmid29363888, year = {2018}, author = {Liu, QQ and Zhang, TS and Li, CX and Gu, JW and Hou, JB and Dong, H}, title = {Decision-making in a bisexual line and a thelytokous Wolbachia-infected line of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) regarding behavior toward their hosts.}, journal = {Pest management science}, volume = {74}, number = {7}, pages = {1720-1727}, doi = {10.1002/ps.4867}, pmid = {29363888}, issn = {1526-4998}, mesh = {Animals ; Decision Making ; Female ; Insect Control ; Moths/*parasitology ; *Pest Control, Biological ; Wasps/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The use of thelytokous Wolbachia-infected Trichogramma (parasitic wasps) has long been considered as a way to enhance the efficacy of biocontrol. However, Wolbachia can affect the host physiology. We compared decision-making between bisexual and thelytokous Wolbachia-infected lines of Trichogramma dendrolimi Matsumura regarding behavior toward fresh and old eggs of Corcyra cephalonica at 25 ± 1 °C and 70 ± 5% relative humidity.

RESULTS: The behavioral patterns and sequences of the two lines were basically the same. The durations of various behavioral patterns and values of fitness indicators of the bisexual line on fresh eggs were generally significantly shorter and better, respectively, than on old eggs, whereas the thelytokous line behaved similarly toward the two types of eggs, and differences in most fitness indicators between fresh and old eggs were not significant. On fresh eggs, the durations of various behaviors in the bisexual line were generally significantly shorter than in the thelytokous line and the fitness indicators were generally significantly better.

CONCLUSION: Wolbachia affected the fitness of T. dendrolimi negatively. The potential of the thelytokous line as a biocontrol agent would not be as good as that of the bisexual line when decision-making only is considered. Therefore, further evaluations need to be carried out before the thelytokous line can be used in practical biocontrol. © 2018 Society of Chemical Industry.}, } @article {pmid29358725, year = {2018}, author = {Schmidt, TL and Filipović, I and Hoffmann, AA and Rašić, G}, title = {Fine-scale landscape genomics helps explain the slow spatial spread of Wolbachia through the Aedes aegypti population in Cairns, Australia.}, journal = {Heredity}, volume = {120}, number = {5}, pages = {386-395}, pmid = {29358725}, issn = {1365-2540}, mesh = {Aedes/*microbiology ; Animals ; Australia ; Female ; Genetics, Population ; *Genomics ; Humans ; Larva ; Male ; Polymorphism, Single Nucleotide/*genetics ; Wolbachia/*genetics/physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia suppresses the capacity for arbovirus transmission in the mosquito Aedes aegypti, and can spread spatially through wild mosquito populations following local introductions. Recent introductions in Cairns, Australia have demonstrated slower than expected spatial spread. Potential reasons for this include: (i) barriers to Ae. aegypti dispersal; (ii) higher incidence of long-range dispersal; and (iii) intergenerational loss of Wolbachia. We investigated these three potential factors using genome-wide single-nucleotide polymorphisms (SNPs) and an assay for the Wolbachia infection wMel in 161 Ae. aegypti collected from Cairns in 2015. We detected a small but significant barrier effect of Cairns highways on Ae. aegypti dispersal using distance-based redundancy analysis and patch-based simulation analysis. We detected a pair of putative full-siblings in ovitraps 1312 m apart, indicating long-distance female movement likely mediated by human transport. We also found a pair of full-siblings of different infection status, indicating intergenerational loss of Wolbachia in the field. These three factors are all expected to contribute to the slow spread of Wolbachia through Ae. aegypti populations, though from our results it is unclear whether Wolbachia loss and long-distance movement are sufficiently common to reduce the speed of spatial spread appreciably. Our findings inform the strategic deployment of Wolbachia-infected mosquitoes during releases, and show how parameter estimates from laboratory studies may differ from those estimated using field data. Our landscape genomics approach can be extended to other host/symbiont systems that are being considered for biocontrol.}, } @article {pmid29358578, year = {2018}, author = {Bonneau, M and Atyame, C and Beji, M and Justy, F and Cohen-Gonsaud, M and Sicard, M and Weill, M}, title = {Culex pipiens crossing type diversity is governed by an amplified and polymorphic operon of Wolbachia.}, journal = {Nature communications}, volume = {9}, number = {1}, pages = {319}, pmid = {29358578}, issn = {2041-1723}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; Crosses, Genetic ; Culex/genetics/*microbiology ; Drosophila melanogaster/genetics/*microbiology ; Female ; *Genome, Bacterial ; Host Specificity ; Male ; *Operon ; Polymorphism, Genetic ; Sequence Alignment ; Sequence Homology, Amino Acid ; Symbiosis/genetics ; Toxin-Antitoxin Systems/genetics ; Wolbachia/*genetics/metabolism ; }, abstract = {Culex pipiens mosquitoes are infected with Wolbachia (wPip) that cause an important diversity of cytoplasmic incompatibilities (CIs). Functional transgenic studies have implicated the cidA-cidB operon from wPip and its homolog in wMel in CI between infected Drosophila males and uninfected females. However, the genetic basis of the CI diversity induced by different Wolbachia strains was unknown. We show here that the remarkable diversity of CI in the C. pipiens complex is due to the presence, in all tested wPip genomes, of several copies of the cidA-cidB operon, which undergoes diversification through recombination events. In 183 isofemale lines of C. pipiens collected worldwide, specific variations of the cidA-cidB gene repertoires are found to match crossing types. The diversification of cidA-cidB is consistent with the hypothesis of a toxin-antitoxin system in which the gene cidB co-diversifies with the gene cidA, particularly in putative domains of reciprocal interactions.}, } @article {pmid29351633, year = {2018}, author = {Lindsey, ARI and Rice, DW and Bordenstein, SR and Brooks, AW and Bordenstein, SR and Newton, ILG}, title = {Evolutionary Genetics of Cytoplasmic Incompatibility Genes cifA and cifB in Prophage WO of Wolbachia.}, journal = {Genome biology and evolution}, volume = {10}, number = {2}, pages = {434-451}, pmid = {29351633}, issn = {1759-6653}, support = {P30 DK058404/DK/NIDDK NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; T32 GM080178/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology/physiology ; Evolution, Molecular ; *Genes, Viral ; Male ; Prophages/*genetics ; Reproduction ; Symbiosis ; Transcriptome ; Wolbachia/genetics/physiology/*virology ; }, abstract = {The bacterial endosymbiont Wolbachia manipulates arthropod reproduction to facilitate its maternal spread through host populations. The most common manipulation is cytoplasmic incompatibility (CI): Wolbachia-infected males produce modified sperm that cause embryonic mortality, unless rescued by embryos harboring the same Wolbachia. The genes underlying CI, cifA and cifB, were recently identified in the eukaryotic association module of Wolbachia's prophage WO. Here, we use transcriptomic and genomic approaches to address three important evolutionary facets of the cif genes. First, we assess whether or not cifA and cifB comprise a classic toxin-antitoxin operon in wMel and show that the two genes exhibit striking, transcriptional differences across host development. They can produce a bicistronic message despite a predicted hairpin termination element in their intergenic region. Second, cifA and cifB strongly coevolve across the diversity of phage WO. Third, we provide new domain and functional predictions across homologs within Wolbachia, and show that amino acid sequences vary substantially across the genus. Finally, we investigate conservation of cifA and cifB and find frequent degradation and loss of the genes in strains that no longer induce CI. Taken together, we demonstrate that cifA and cifB exhibit complex transcriptional regulation in wMel, provide functional annotations that broaden the potential mechanisms of CI induction, and report recurrent erosion of cifA and cifB in non-CI strains, thus expanding our understanding of the most widespread form of reproductive parasitism.}, } @article {pmid29348597, year = {2018}, author = {Aljayyoussi, G and Tyrer, HE and Ford, L and Sjoberg, H and Pionnier, N and Waterhouse, D and Davies, J and Gamble, J and Metuge, H and Cook, DAN and Steven, A and Sharma, R and Guimaraes, AF and Clare, RH and Cassidy, A and Johnston, KL and Myhill, L and Hayward, L and Wanji, S and Turner, JD and Taylor, MJ and Ward, SA}, title = {Author Correction: Short-Course, High-Dose Rifampicin Achieves Wolbachia Depletion Predictive of Curative Outcomes in Preclinical Models of Lymphatic Filariasis and Onchocerciasis.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {1384}, doi = {10.1038/s41598-018-19723-1}, pmid = {29348597}, issn = {2045-2322}, abstract = {A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.}, } @article {pmid29346449, year = {2018}, author = {Chrostek, E and Teixeira, L}, title = {Within host selection for faster replicating bacterial symbionts.}, journal = {PloS one}, volume = {13}, number = {1}, pages = {e0191530}, pmid = {29346449}, issn = {1932-6203}, mesh = {Animals ; Drosophila melanogaster/*microbiology ; Genes, Bacterial ; *Host-Pathogen Interactions ; *Symbiosis ; Virulence/genetics ; Wolbachia/genetics/*growth & development/pathogenicity ; }, abstract = {Wolbachia is a widespread, intracellular symbiont of arthropods, able to induce reproductive distortions and antiviral protection in insects. Wolbachia can also be pathogenic, as is the case with wMelPop, a virulent variant of the endosymbiont of Drosophila melanogaster. An extensive genomic amplification of the 20kb region encompassing eight Wolbachia genes, called Octomom, is responsible for wMelPop virulence. The Octomom copy number in wMelPop can be highly variable between individual D. melanogaster flies, even when comparing siblings arising from a single female. Moreover, Octomom copy number can change rapidly between generations. These data suggest an intra-host variability in Octomom copy number between Wolbachia cells. Since wMelPop Wolbachia with different Octomom copy numbers grow at different rates, we hypothesized that selection could act on this intra-host variability. Here we tested if total Octomom copy number changes during the lifespan of individual Drosophila hosts, revealing selection for different Wolbachia populations. We performed a time course analysis of Octomom amplification in flies whose mothers were controlled for Octomom copy number. We show that despite the Octomom copy number being relatively stable it increases slightly throughout D. melanogaster adult life. This indicates that there is selection acting on the intra-host variation in the Octomom copy number over the lifespan of individual hosts. This within host selection for faster replicating bacterial symbionts may be in conflict with between host selection against highly pathogenic Wolbachia.}, } @article {pmid29336504, year = {2018}, author = {Osei-Amo, S and Hussein, M and Asad, S and Hugo, L and Asgari, S}, title = {Wolbachia-induced transcription factor GATA4 suppresses ovary-specific genes blastoderm-specific protein 25D and imaginal disc growth factor.}, journal = {Insect molecular biology}, volume = {27}, number = {3}, pages = {295-304}, doi = {10.1111/imb.12371}, pmid = {29336504}, issn = {1365-2583}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Blastoderm/growth & development ; Female ; GATA4 Transcription Factor/*genetics/metabolism ; Imaginal Discs/growth & development ; Insect Proteins/*genetics/metabolism ; Ovary/metabolism ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia infects a wide array of insect hosts and has been implicated in a range of biological modifications as a consequence of its infection. Previously, it was shown that the transcription factor GATA4 was significantly induced in Wolbachia wMelPop-CLA strain infected Aedes aegypti whole mosquitoes and cells. Here, we provide evidence that this induction also occurs in mosquito ovaries where the ovary-specific genes blastoderm-specific protein 25D (Bsg25D) and imaginal disc growth factor (Disc) are suppressed by Wolbachia. We further demonstrate that transcriptional depletion of GATA4 results in upregulation of both genes and conversely its overexpression leads to downregulation of the genes, suggesting that Wolbachia-induced GATA4 plays a suppressive regulatory role with regards to Bsg25D and Disc expression in mosquito ovaries. When the Disc gene was silenced in mosquitoes, we did not observe any difference in the number of mature ovarian follicles developed between treatment groups. However, we did find a significant delay in the hatching of eggs that had been laid by Disc knockdown mosquitoes.}, } @article {pmid29334340, year = {2019}, author = {Amala, M and Rajamanikandan, S and Prabhu, D and Surekha, K and Jeyakanthan, J}, title = {Identification of anti-filarial leads against aspartate semialdehyde dehydrogenase of Wolbachia endosymbiont of Brugia malayi: combined molecular docking and molecular dynamics approaches.}, journal = {Journal of biomolecular structure & dynamics}, volume = {37}, number = {2}, pages = {394-410}, doi = {10.1080/07391102.2018.1427633}, pmid = {29334340}, issn = {1538-0254}, mesh = {Amino Acid Sequence ; Animals ; Anthelmintics/*chemistry/pharmacology ; Aspartate-Semialdehyde Dehydrogenase/antagonists & inhibitors/*chemistry ; Binding Sites ; Brugia malayi/*enzymology ; Catalytic Domain ; Chemical Phenomena ; Drug Discovery/methods ; Enzyme Inhibitors/*chemistry/pharmacology ; Humans ; Hydrogen Bonding ; Hydrophobic and Hydrophilic Interactions ; *Molecular Docking Simulation ; *Molecular Dynamics Simulation ; Protein Binding ; }, abstract = {Lymphatic filariasis is a debilitating vector borne parasitic disease that infects human lymphatic system by nematode Brugia malayi. Currently available anti-filarial drugs are effective only on the larval stages of parasite. So far, no effective drugs are available for humans to treat filarial infections. In this regard, aspartate semialdehyde dehydrogenase (ASDase) in lysine biosynthetic pathway from Wolbachia endosymbiont Brugia malayi represents an attractive therapeutic target for the development of novel anti-filarial agents. In this present study, molecular modeling combined with molecular dynamics simulations and structure-based virtual screening were performed to identify potent lead molecules against ASDase. Based on Glide score, toxicity profile, binding affinity and mode of interactions with the ASDase, five potent lead molecules were selected. The molecular docking and dynamics results revealed that the amino acid residues Arg103, Asn133, Cys134, Gln161, Ser164, Lys218, Arg239, His246, and Asn321 plays a crucial role in effective binding of Top leads into the active site of ASDase. The stability of the ASDase-lead complexes was confirmed by running the 30 ns molecular dynamics simulations. The pharmacokinetic properties of the identified lead molecules are in the acceptable range. Furthermore, density functional theory and binding free energy calculations were performed to rank the lead molecules. Thus, the identified lead molecules can be used for the development of anti-filarial agents to combat the pathogenecity of Brugia malayi.}, } @article {pmid29330177, year = {2018}, author = {Zhu, YX and Song, YL and Zhang, YK and Hoffmann, AA and Zhou, JC and Sun, JT and Hong, XY}, title = {Incidence of Facultative Bacterial Endosymbionts in Spider Mites Associated with Local Environments and Host Plants.}, journal = {Applied and environmental microbiology}, volume = {84}, number = {6}, pages = {}, pmid = {29330177}, issn = {1098-5336}, mesh = {Animals ; Bacteroidetes/*physiology ; China ; Environment ; Solanum lycopersicum/growth & development ; Glycine max/growth & development ; Spiroplasma/*physiology ; *Symbiosis ; Tetranychidae/*microbiology/physiology ; Wolbachia/*physiology ; Zea mays/growth & development ; }, abstract = {Spider mites are frequently associated with multiple endosymbionts whose infection patterns often exhibit spatial and temporal variation. However, the association between endosymbiont prevalence and environmental factors remains unclear. Here, we surveyed endosymbionts in natural populations of the spider mite, Tetranychus truncatus, in China, screening 935 spider mites from 21 localities and 12 host plant species. Three facultative endosymbiont lineages, Wolbachia, Cardinium, and Spiroplasma, were detected at different infection frequencies (52.5%, 26.3%, and 8.6%, respectively). Multiple endosymbiont infections were observed in most local populations, and the incidence of individuals with the Wolbachia-Spiroplasma coinfection was higher than expected from the frequency of each infection within a population. Endosymbiont infection frequencies exhibited associations with environmental factors: Wolbachia infection rates increased at localities with higher annual mean temperatures, while Cardinium and Spiroplasma infection rates increased at localities from higher altitudes. Wolbachia was more common in mites from Lycopersicon esculentum and Glycine max compared to those from Zea mays This study highlights that host-endosymbiont interactions may be associated with environmental factors, including climate and other geographically linked factors, as well as the host's food plant.IMPORTANCE The aim of this study was to examine the incidence of endosymbiont distribution and the infection patterns in spider mites. The main findings are that multiple endosymbiont infections were more common than expected and that endosymbiont infection frequencies were associated with environmental factors. This work highlights that host-endosymbiont interactions need to be studied within an environmental and geographic context.}, } @article {pmid29311247, year = {2018}, author = {Mann, M and Fattah-Hosseini, S and Ammar, ED and Stange, R and Warrick, E and Sturgeon, K and Shatters, R and Heck, M}, title = {Diaphorina citri Nymphs Are Resistant to Morphological Changes Induced by "Candidatus Liberibacter asiaticus" in Midgut Epithelial Cells.}, journal = {Infection and immunity}, volume = {86}, number = {4}, pages = {}, pmid = {29311247}, issn = {1098-5522}, mesh = {Animals ; Epithelial Cells/*microbiology ; Female ; Gastrointestinal Microbiome ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Microscopy, Confocal ; Nymph/*microbiology ; Oxidative Stress ; Phenotype ; Rhizobiaceae/*physiology ; }, abstract = {"Candidatus Liberibacter asiaticus" is the causative bacterium associated with citrus greening disease. "Ca Liberibacter asiaticus" is transmitted by Diaphorina citri more efficiently when it is acquired by nymphs rather than adults. Why this occurs is not known. We compared midguts of D. citri insects reared on healthy or "Ca Liberibacter asiaticus"-infected citrus trees using quantitative PCR, confocal microscopy, and mitochondrial superoxide staining for evidence of oxidative stress. Consistent with its classification as propagative, "Ca Liberibacter asiaticus" titers were higher in adults than in nymphs. Our previous work showed that adult D. citri insects have basal levels of karyorrhexis (fragmentation of the nucleus) in midgut epithelial cells, which is increased in severity and frequency in response to "Ca Liberibacter asiaticus." Here, we show that nymphs exhibit lower levels of early-stage karyorrhexis than adults and are refractory to the induction of advanced karyorrhexis by "Ca Liberibacter asiaticus" in the midgut epithelium. MitoSox Red staining showed that guts of infected adults, particularly males, experienced oxidative stress in response to "Ca Liberibacter asiaticus." A positive correlation between the titers of "Ca Liberibacter asiaticus" and the Wolbachia endosymbiont was observed in adult and nymph midguts, suggesting an interplay between these bacteria during development. We hypothesize that the resistance of the nymph midgut to late-stage karyorrhexis through as yet unknown molecular mechanisms benefits "Ca Liberibacter asiaticus" for efficient invasion of midgut epithelial cells, which may be a factor explaining the developmental dependency of "Ca Liberibacter asiaticus" acquisition by the vector.}, } @article {pmid29308790, year = {2018}, author = {Sharma, R and Aljayyoussi, G and Tyrer, HE and Gamble, J and Hayward, L and Guimaraes, AF and Davies, J and Waterhouse, D and Cook, DAN and Myhill, LJ and Clare, RH and Cassidy, A and Steven, A and Johnston, KL and Ford, L and Turner, JD and Ward, SA and Taylor, MJ}, title = {Corrigendum: Minocycline as a re-purposed anti-Wolbachia macrofilaricide: superiority compared with doxycycline regimens in a murine infection model of human lymphatic filariasis.}, journal = {Scientific reports}, volume = {8}, number = {}, pages = {46934}, doi = {10.1038/srep46934}, pmid = {29308790}, issn = {2045-2322}, abstract = {This corrects the article DOI: 10.1038/srep23458.}, } @article {pmid29303740, year = {2018}, author = {De Ambrogi, M}, title = {Highlights from the 66th annual ASTMH meeting.}, journal = {The Lancet. Infectious diseases}, volume = {18}, number = {1}, pages = {32}, doi = {10.1016/S1473-3099(17)30717-X}, pmid = {29303740}, issn = {1474-4457}, mesh = {Antimalarials/*therapeutic use ; Arbovirus Infections/*prevention & control ; Disease Eradication ; *Disease Management ; Drug Therapy, Combination/methods ; Global Health ; HIV Infections/*drug therapy/prevention & control ; Humans ; Lassa Fever/diagnosis/*pathology/therapy ; Malaria/*drug therapy ; Mosquito Control/*methods ; Randomized Controlled Trials as Topic ; Treatment Outcome ; Wolbachia/growth & development ; }, } @article {pmid29301977, year = {2018}, author = {Dunning Hotopp, JC and Klasson, L}, title = {The Complexities and Nuances of Analyzing the Genome of Drosophila ananassae and Its Wolbachia Endosymbiont.}, journal = {G3 (Bethesda, Md.)}, volume = {8}, number = {1}, pages = {373-374}, pmid = {29301977}, issn = {2160-1836}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; }, mesh = {Animals ; Drosophila/genetics ; Drosophila melanogaster ; Genome ; Retroelements ; Wolbachia/*genetics ; }, abstract = {In "Retrotransposons Are the Major Contributors to the Expansion of the Drosophila ananassae Muller F Element," Leung et al. (2017) improved contigs attributed to the Muller F element from the original CAF1 assembly, and used them to conclude that most of the sequence expansion of the fourth chromosome of D. ananassae is due to a higher transposon load than previously thought, but is not due to Wolbachia DNA integrations. While we do not disagree with the first conclusion, the authors base their second conclusion on the lack of homology detected between their improved CAF1 genome assembly attributed to D. ananassae and reference Wolbachia genomes. While the consensus CAF1 genome assembly lacks any sequence similarity to the reference genome of the Wolbachia endosymbiont of Drosophila melanogaster (wMel), numerous studies from multiple laboratories provide experimental support for a large lateral/horizontal gene transfer (LGT) of a Wolbachia genome into this D. ananassae line. As such, we strongly suspect that the original whole genome assembly was either constructed after the removal of all Wolbachia reads, or that Wolbachia sequences were directly removed from the contigs in the CAF1 assembly. Hence, Leung et al. (2017) could not have identified the Wolbachia LGT using the CAF1 assembly. This manuscript by Leung et al. (2017) highlights that an assembly of the Wolbachia sequence reads and their mate pairs was erroneously attributed solely to the Wolbachia endosymbiont, albeit before we understood the extent of LGT in D. ananassae As such, we recommend that the sequences deposited at the National Center for Biotechnology Information (NCBI) under PRJNA13365 should not be attributed to Wolbachia endosymbiont of D. ananassae, but should have their taxonomy reclassified by NCBI as "Unclassified sequences." As our knowledge about genome biology improves, we need to reconsider and reanalyze earlier genomes removing the prejudice introduced from now defunct paradigms.}, } @article {pmid29300732, year = {2018}, author = {Specht, S and Pfarr, KM and Arriens, S and Hübner, MP and Klarmann-Schulz, U and Koschel, M and Sternberg, S and Martin, C and Ford, L and Taylor, MJ and Hoerauf, A}, title = {Combinations of registered drugs reduce treatment times required to deplete Wolbachia in the Litomosoides sigmodontis mouse model.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {1}, pages = {e0006116}, pmid = {29300732}, issn = {1935-2735}, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage ; Disease Models, Animal ; Drug Therapy, Combination/methods ; Filariasis/*drug therapy ; Filarioidea/*microbiology ; Fluoroquinolones/administration & dosage ; Mice ; Moxifloxacin ; Rifampin/administration & dosage/analogs & derivatives ; Tetracyclines/administration & dosage ; Time Factors ; Treatment Outcome ; Wolbachia/*drug effects ; }, abstract = {Filarial parasites can be targeted by antibiotic treatment due to their unique endosymbiotic relationship with Wolbachia bacteria. This finding has led to successful treatment strategies in both, human onchocerciasis and lymphatic filariasis. A 4-6 week treatment course using doxycycline results in long-term sterility and safe macrofilaricidal activity in humans. However, current treatment times and doxycycline contraindications in children and pregnant women preclude widespread administration of doxycycline in public health control programs; therefore, the search for shorter anti-wolbachial regimens is a focus of ongoing research. We have established an in vivo model for compound screening, using mice infected with Litomosoides sigmodontis. We could show that gold standard doxycycline treatment did not only deplete Wolbachia, it also resulted in a larval arrest. In this model, combinations of registered antibiotics were tested for their anti-wolbachial activity. Administration of rifamycins in combination with doxycycline for 7 days successfully depleted Wolbachia by > 2 log (>99% reduction) and thus resulted in a significant reduction of the treatment duration. Using a triple combination of a tetracycline (doxycycline or minocycline), a rifamycin and a fluoroquinolone (moxifloxacin) led to an even greater shortening of the treatment time. Testing all double combinations that could be derived from the triple combinations revealed that the combination of rifapentine (15mg/kg) and moxifloxacin (2 x 200mg/kg) showed the strongest reduction of treatment time in intraperitoneal and also oral administration routes. The rifapentine plus moxifloxacin combination was equivalent to the triple combination with additional doxycycline (>99% Wolbachia reduction). These investigations suggest that it is possible to shorten anti-wolbachial treatment times with combination treatments in order to achieve the target product profile (TPP) requirements for macrofilaricidal drugs of no more than 7-10 days of treatment.}, } @article {pmid29297702, year = {2018}, author = {Schultz, MJ and Connor, JH and Frydman, HM}, title = {Group B Wolbachia Strain-Dependent Inhibition of Arboviruses.}, journal = {DNA and cell biology}, volume = {37}, number = {1}, pages = {2-6}, pmid = {29297702}, issn = {1557-7430}, mesh = {Aedes/virology ; Animals ; Arboviruses/*physiology ; Culicidae/virology ; Dengue Virus/physiology ; Phylogeny ; Virus Replication/physiology ; Wolbachia/*virology ; Zika Virus/physiology ; }, abstract = {Mosquito-borne viruses, including Zika virus (ZIKV) and dengue virus (DENV), are global threats that continue to infect millions annually. Historically, efforts to combat the spread of these diseases have sought to eradicate the mosquito population. This has had limited success. Recent efforts to combat the spread of these diseases have targeted the mosquito population and the mosquito's ability to transmit viruses by altering the mosquito's microbiome. The introduction of particular strains of Wolbachia bacteria into mosquitos suppresses viral growth and blocks disease transmission. This novel strategy is being tested worldwide to reduce DENV and has early indications of success. The Wolbachia genus comprised divergent strains that are divided in major phylogenetic clades termed supergroups. All Wolbachia field trials currently utilize supergroup A Wolbachia in Aedes aegypti mosquitos to limit virus transmission. Here we discuss our studies of Wolbachia strains not yet used in virus control strategies but that show strong potential to reduce ZIKV replication. These strains are important opportunities in the search for novel tools to reduce the levels of mosquito-borne viruses and provide additional models for mechanistic studies.}, } @article {pmid29297293, year = {2017}, author = {Gruntenko, NЕ and Ilinsky, YY and Adonyeva, NV and Burdina, EV and Bykov, RA and Menshanov, PN and Rauschenbach, IY}, title = {Various Wolbachia genotypes differently influence host Drosophila dopamine metabolism and survival under heat stress conditions.}, journal = {BMC evolutionary biology}, volume = {17}, number = {Suppl 2}, pages = {252}, pmid = {29297293}, issn = {1471-2148}, mesh = {Alkaline Phosphatase/metabolism ; Animals ; Arylalkylamine N-Acetyltransferase/metabolism ; Dihydroxyphenylalanine/pharmacology ; Dopamine/*metabolism ; Drosophila Proteins/metabolism ; Drosophila melanogaster/enzymology/genetics/*microbiology ; Female ; Genotype ; *Hot Temperature ; Male ; *Stress, Physiological/drug effects ; Survival Analysis ; Wolbachia/drug effects/*genetics ; }, abstract = {BACKGROUND: One of the most widespread prokaryotic symbionts of invertebrates is the intracellular bacteria of Wolbachia genus which can be found in about 50% of insect species. Wolbachia causes both parasitic and mutualistic effects on its host that include manipulating the host reproductive systems in order to increase their transmission through the female germline, and increasing the host fitness. One of the mechanisms, promoting adaptation in biological organisms, is a non-specific neuroendocrine stress reaction. In insects, this reaction includes catecholamines, dopamine, serotonin and octopamine, which act as neurotransmitters, neuromodulators and neurohormones. The level of dopamine metabolism correlates with heat stress resistance in Drosophila adults.

RESULTS: To examine Wolbachia effect on Drosophila survival under heat stress and dopamine metabolism we used five strains carrying the nuclear background of interbred Bi90 strain and cytoplasmic backgrounds with different genotype variants of Wolbachia (produced by 20 backcrosses of Bi90 males with appropriate source of Wolbachia). Non-infected Bi90 strain (treated with tetracycline for 3 generations) was used as a control group. We demonstrated that two of five investigated Wolbachia variants promote changes in Drosophila heat stress resistance and activity of enzymes that produce and degrade dopamine, alkaline phosphatase and dopamine-dependent arylalkylamine N-acetyltransferase. What is especially interesting, wMelCS genotype of Wolbachia increases stress resistance and the intensity of dopamine metabolism, whereas wMelPop strain decreases them. wMel, wMel2 and wMel4 genotypes of Wolbachia do not show any effect on the survival under heat stress or dopamine metabolism. L-DOPA treatment, known to increase the dopamine content in Drosophila, levels the difference in survival under heat stress between all studied groups.

CONCLUSIONS: The genotype of symbiont determines the effect that the symbiont has on the stress resistance of the host insect.}, } @article {pmid29294302, year = {2018}, author = {Liu, T and Xu, Y and Wang, X and Gu, J and Yan, G and Chen, XG}, title = {Antiviral systems in vector mosquitoes.}, journal = {Developmental and comparative immunology}, volume = {83}, number = {}, pages = {34-43}, doi = {10.1016/j.dci.2017.12.025}, pmid = {29294302}, issn = {1879-0089}, mesh = {Animals ; Arboviruses/*physiology ; Culicidae/*immunology/virology ; Cytokines/metabolism ; Gastrointestinal Microbiome ; Humans ; Insect Proteins/metabolism ; Mammals ; Mosquito Vectors/*virology ; Pathogen-Associated Molecular Pattern Molecules/immunology ; RNA Interference ; RNA, Small Interfering/genetics ; Receptors, Pattern Recognition/metabolism ; Signal Transduction ; Virus Replication ; }, abstract = {Mosquito-borne viral diseases represent a major challenge to human public health. As natural vectors of arboviruses, mosquitoes can be infected by a virus, but they have evolved multiple mechanisms to tolerate constant infection and restrict viral replication via their antiviral immune system. In a state of continuous infection, a mosquito can transmit an arbovirus while obtaining a blood meal from a mammalian host. During infection, the virus is mainly inhibited through a small RNA-mediated interference mechanism. Within mosquitoes, the invaded viruses are recognized based on pathogen-associated molecular patterns, leading to the production of cytokines. These cytokines in turn bind pattern recognition receptors and activate Toll, IMD and other immune signalling pathways to expand the immune response and induce antiviral activity via immune effectors. Interestingly, the gut microbiota and Wolbachia also play a role in mosquito antiviral immunity, which is very similar to acquired immunity. This review describes the advances made in understanding various aspects of mosquito antiviral immune molecular mechanisms in detail and explores some of the unresolved issues related to the mosquito immune system.}, } @article {pmid29287072, year = {2017}, author = {Baughman, T and Peterson, C and Ortega, C and Preston, SR and Paton, C and Williams, J and Guy, A and Omodei, G and Johnson, B and Williams, H and O'Neill, SL and Ritchie, SA and Dobson, SL and Madan, D}, title = {A highly stable blood meal alternative for rearing Aedes and Anopheles mosquitoes.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {12}, pages = {e0006142}, pmid = {29287072}, issn = {1935-2735}, mesh = {Adenosine Triphosphate/blood/*pharmacology ; Aedes/drug effects/microbiology/*physiology ; Animals ; Anopheles/drug effects/microbiology/*physiology ; Blood Substitutes/chemistry ; Diet ; Dietary Supplements ; Female ; Insect Vectors/drug effects/microbiology/*physiology ; Male ; Ovum ; Pest Control, Biological ; Plasma/*chemistry ; Reproduction/drug effects ; Wolbachia/*physiology ; }, abstract = {We investigated alternatives to whole blood for blood feeding of mosquitoes with a focus on improved stability and compatibility with mass rearing programs. In contrast to whole blood, an artificial blood diet of ATP-supplemented plasma was effective in maintaining mosquito populations and was compatible with storage for extended periods refrigerated, frozen, and as a lyophilized powder. The plasma ATP diet supported rearing of both Anopheles and Aedes mosquitoes. It was also effective in rearing Wolbachia-infected Aedes mosquitoes, suggesting compatibility with vector control efforts.}, } @article {pmid29286204, year = {2019}, author = {Chu, CC and Hoffmann, M and Braswell, WE and Pelz-Stelinski, KS}, title = {Genetic variation and potential coinfection of Wolbachia among widespread Asian citrus psyllid (Diaphorina citri Kuwayama) populations.}, journal = {Insect science}, volume = {26}, number = {4}, pages = {671-682}, pmid = {29286204}, issn = {1744-7917}, support = {#2012-51181-20086//United States Department of Agriculture, National Institute for Food and Agriculture Specialty Crop Research/ ; }, mesh = {Animals ; Coinfection ; Genes, Bacterial ; Genes, Insect ; Genes, Mitochondrial ; Genetic Variation ; Geography ; Hemiptera/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia can profoundly influence the survival, reproduction, and defenses of insect hosts. These interactions could potentially be harnessed for managing pests or insect-transmitted diseases. Diaphorina citri Kuwayama is a phloem-feeding pest capable of transmitting the putative causal agent of citrus greening, Candidatus Liberibacter asiaticus (CLas). Like many insects, D. citri is also infected with Wolbachia (wDi). Recent studies indicate that the relative abundance of wDi could be associated with the abundance of CLas, and that wDi may contribute to regulating expression of phage lytic cycle genes in CLas, suggesting the need for better understanding of wDi biology in general. This study investigated the genetic diversity of wDi among D. citri in populations spanning eleven countries and two U.S. territories. Six Wolbachia genes, wsp, coxA, fbpA, ftsZ, gatB, and hcpA, were sequenced and compared across samples. Two prevalent wDi strains were identified across the samples, and screening of clone libraries revealed possible coinfection of wDi strains in specific populations. D. citri mitochondrial cytochrome oxidase subunit I gene (mtCOI) were more divergent between D. citri populations that were infected with different wDi strains or had different infection statuses (single infection vs. coinfection). While we could not eliminate the possibility that maternal transmission may contribute to such patterns, it is also possible that wDi may induce cytoplasmic incompatibility in their host. These findings should contribute to the understanding of wDi population ecology, which may facilitate manipulation of this endosymbiont for management of citrus greening disease worldwide.}, } @article {pmid29282144, year = {2017}, author = {Terradas, G and Allen, SL and Chenoweth, SF and McGraw, EA}, title = {Family level variation in Wolbachia-mediated dengue virus blocking in Aedes aegypti.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {622}, pmid = {29282144}, issn = {1756-3305}, support = {APP1103804//National Health and Medical Research Council/International ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; *Antibiosis ; Dengue Virus/*growth & development/isolation & purification ; Viral Load ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The mosquito vector Aedes aegypti is responsible for transmitting a range of arboviruses including dengue (DENV) and Zika (ZIKV). The global reach of these viruses is increasing due to an expansion of the mosquito's geographic range and increasing urbanization and human travel. Vector control remains the primary means for limiting these diseases. Wolbachia pipientis is an endosymbiotic bacterium of insects that has the ability to block the replication of pathogens, including flaviviruses such as DENV or ZIKV, inside the body of the vector. A strain of Wolbachia called wMel is currently being released into wild mosquito populations to test its potential to limit virus transmission to humans. The mechanism that underpins the virus blocking effect, however, remains elusive.

METHODS: We used a modified full-sib breeding design in conjunction with vector competence assays in wildtype and wMel-infected Aedes aegypti collected from the field. All individuals were injected with DENV-2 intrathoracically at 5-6 days of age. Tissues were dissected 7 days post-infection to allow quantification of DENV and Wolbachia loads.

RESULTS: We show the first evidence of family level variation in Wolbachia-mediated blocking in mosquitoes. This variation may stem from either genetic contributions from the mosquito and Wolbachia genomes or environmental influences on Wolbachia. In these families, we also tested for correlations between strength of blocking and expression level for several insect immunity genes with possible roles in blocking, identifying two genes of interest (AGO2 and SCP-2).

CONCLUSIONS: In this study we show variation in Wolbachia-mediated DENV blocking in Aedes aegypti that may arise from genetic contributions and environmental influences on the mosquito-Wolbachia association. This suggests that Wolbachia-mediated blocking may have the ability to evolve through time or be expressed differentially across environments. The long-term efficacy of Wolbachia in the field will be dependent on the stability of blocking. Understanding the mechanism of blocking will be necessary for successful development of strategies that counter the emergence of evolved resistance or variation in its expression under diverse field conditions.}, } @article {pmid29279375, year = {2018}, author = {Carrington, LB and Tran, BCN and Le, NTH and Luong, TTH and Nguyen, TT and Nguyen, PT and Nguyen, CVV and Nguyen, HTC and Vu, TT and Vo, LT and Le, DT and Vu, NT and Nguyen, GT and Luu, HQ and Dang, AD and Hurst, TP and O'Neill, SL and Tran, VT and Kien, DTH and Nguyen, NM and Wolbers, M and Wills, B and Simmons, CP}, title = {Field- and clinically derived estimates of Wolbachia-mediated blocking of dengue virus transmission potential in Aedes aegypti mosquitoes.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {115}, number = {2}, pages = {361-366}, pmid = {29279375}, issn = {1091-6490}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/microbiology/*virology ; Animals ; Dengue/blood/transmission/*virology ; Dengue Virus/*physiology ; Humans ; Logistic Models ; Mosquito Vectors/microbiology/*virology ; Pest Control, Biological/methods ; Time Factors ; Viremia/blood/virology ; Wolbachia/*physiology ; }, abstract = {The wMel strain of Wolbachia can reduce the permissiveness of Aedes aegypti mosquitoes to disseminated arboviral infections. Here, we report that wMel-infected Ae. aegypti (Ho Chi Minh City background), when directly blood-fed on 141 viremic dengue patients, have lower dengue virus (DENV) transmission potential and have a longer extrinsic incubation period than their wild-type counterparts. The wMel-infected mosquitoes that are field-reared have even greater relative resistance to DENV infection when fed on patient-derived viremic blood meals. This is explained by an increased susceptibility of field-reared wild-type mosquitoes to infection than laboratory-reared counterparts. Collectively, these field- and clinically relevant findings support the continued careful field-testing of wMel introgression for the biocontrol of Ae. aegypti-born arboviruses.}, } @article {pmid29274498, year = {2018}, author = {Kajtoch, Ł and Montagna, M and Wanat, M}, title = {Species delimitation within the Bothryorrhynchapion weevils: Multiple evidence from genetics, morphology and ecological associations.}, journal = {Molecular phylogenetics and evolution}, volume = {120}, number = {}, pages = {354-363}, doi = {10.1016/j.ympev.2017.12.022}, pmid = {29274498}, issn = {1095-9513}, mesh = {Animals ; DNA/chemistry/isolation & purification/metabolism ; Electron Transport Complex IV/classification/genetics ; Female ; Genetic Variation ; Male ; Peptide Elongation Factor 1/classification/genetics ; Phylogeny ; Sequence Analysis, DNA ; Weevils/*classification/genetics/parasitology ; Wolbachia/genetics/physiology ; }, abstract = {Curculionidae is a hyperdiverse group of beetles, whose taxonomy and phylogenetics are still poorly understood, especially at the genus level. The latest work on the evolution of Apionini showed a noticeable "mess" in the subtribe Oxystomatina, where most of the morphology-based genera were found to be polyphyletic or paraphyletic. These discrepancies between classical taxonomy and molecular phylogenetics implied the need for further taxonomic revision of these groups. Here, we used sets of morphological, molecular and ecological characters to verify the taxonomic statuses and disentangle the phylogenetic relations among the Bothryorrhynchapion apionids, which are classified as a subgenus of Cyanapion. Morphological data including morphometrics, and multilocus molecular analyses confirmed the monophyly of the Bothryorrhynchapion and species statuses of five species. The morphological analyses showed that Cyanapion (Bothryorrhynchapion) protractum (Sharp, 1891) from the southeast Palaearctic is a synonym of C. (B.) gyllenhalii (Kirby). Moreover, ecological features (host plant use and presence/absence of the endosymbiotic bacteria Wolbachia) helped to unravel the relations among the examined weevils. The speciation of Bothryorrhynchapion apionids was probably affected by allopatric distribution, shifts in the preferred host plants (Vicia sp. or Lathyrus sp.) of sympatric taxa, and infection by different strains of Wolbachia. The paper presents the first comprehensive description of the species' morphology, biology and ecology, and includes a key to the species.}, } @article {pmid29257089, year = {2017}, author = {Poff, KE and Stever, H and Reil, JB and Seabourn, P and Ching, AJ and Aoki, S and Logan, M and Michalski, JR and Santamaria, J and Adams, JW and Eiben, JA and Yew, JY and Ewing, CP and Magnacca, KN and Bennett, GM}, title = {The Native Hawaiian Insect Microbiome Initiative: A Critical Perspective for Hawaiian Insect Evolution.}, journal = {Insects}, volume = {8}, number = {4}, pages = {}, pmid = {29257089}, issn = {2075-4450}, abstract = {Insects associate with a diversity of microbes that can shape host ecology and diversity by providing essential biological and adaptive services. For most insect groups, the evolutionary implications of host-microbe interactions remain poorly understood. Geographically discrete areas with high biodiversity offer powerful, simplified model systems to better understand insect-microbe interactions. Hawaii boasts a diverse endemic insect fauna (~6000 species) characterized by spectacular adaptive radiations. Despite this, little is known about the role of bacteria in shaping this diversity. To address this knowledge gap, we inaugurate the Native Hawaiian Insect Microbiome Initiative (NHIMI). The NHIMI is an effort intended to develop a framework for informing evolutionary and biological studies in Hawaii. To initiate this effort, we have sequenced the bacterial microbiomes of thirteen species representing iconic, endemic Hawaiian insect groups. Our results show that native Hawaiian insects associate with a diversity of bacteria that exhibit a wide phylogenetic breadth. Several groups show predictable associations with obligate microbes that permit diet specialization. Others exhibit unique ecological transitions that are correlated with shifts in their microbiomes (e.g., transition to carrion feeding from plant-feeding in Nysius wekiuicola). Finally, some groups, such as the Hawaiian Drosophila, have relatively diverse microbiomes with a conserved core of bacterial taxa across multiple species and islands.}, } @article {pmid29250466, year = {2017}, author = {Šochová, E and Husník, F and Nováková, E and Halajian, A and Hypša, V}, title = {Arsenophonus and Sodalis replacements shape evolution of symbiosis in louse flies.}, journal = {PeerJ}, volume = {5}, number = {}, pages = {e4099}, pmid = {29250466}, issn = {2167-8359}, abstract = {Symbiotic interactions between insects and bacteria are ubiquitous and form a continuum from loose facultative symbiosis to greatly intimate and stable obligate symbiosis. In blood-sucking insects living exclusively on vertebrate blood, obligate endosymbionts are essential for hosts and hypothesized to supplement B-vitamins and cofactors missing from their blood diet. The role and distribution of facultative endosymbionts and their evolutionary significance as seeds of obligate symbioses are much less understood. Here, using phylogenetic approaches, we focus on the Hippoboscidae phylogeny as well as the stability and dynamics of obligate symbioses within this bloodsucking group. In particular, we demonstrate a new potentially obligate lineage of Sodalis co-evolving with the Olfersini subclade of Hippoboscidae. We also show several likely facultative Sodalis lineages closely related to Sodalis praecaptivus (HS strain) and suggest repeated acquisition of novel symbionts from the environment. Similar to Sodalis, Arsenophonus endosymbionts also form both obligate endosymbiotic lineages co-evolving with their hosts (Ornithomyini and Ornithoica groups) as well as possibly facultative infections incongruent with the Hippoboscidae phylogeny. Finally, we reveal substantial diversity of Wolbachia strains detected in Hippoboscidae samples falling into three supergroups: A, B, and the most common F. Altogether, our results prove the associations between Hippoboscoidea and their symbiotic bacteria to undergo surprisingly dynamic, yet selective, evolutionary processes strongly shaped by repeated endosymbiont replacements. Interestingly, obligate symbionts only originate from two endosymbiont genera, Arsenophonus and Sodalis, suggesting that the host is either highly selective about its future obligate symbionts or that these two lineages are the most competitive when establishing symbioses in louse flies.}, } @article {pmid29248525, year = {2018}, author = {Huang, M and Luo, J and Hu, L and Zheng, B and Yu, J}, title = {Assessing the efficiency of Wolbachia driven Aedes mosquito suppression by delay differential equations.}, journal = {Journal of theoretical biology}, volume = {440}, number = {}, pages = {1-11}, doi = {10.1016/j.jtbi.2017.12.012}, pmid = {29248525}, issn = {1095-8541}, mesh = {Aedes/*microbiology/parasitology ; Animals ; Female ; Infertility/*microbiology ; Male ; Models, Biological ; *Models, Theoretical ; Mosquito Vectors ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, abstract = {To suppress wild population of Aedes mosquitoes, the primary transmission vector of life-threatening diseases such as dengue, malaria, and Zika, an innovative strategy is to release male mosquitoes carrying the bacterium Wolbachia into natural areas to drive female sterility by cytoplasmic incompatibility. We develop a model of delay differential equations, incorporating the strong density restriction in the larval stage, to assess the delicate impact of life table parameters on suppression efficiency. Through mathematical analysis, we find the sufficient and necessary condition for global stability of the complete suppression state. This condition, combined with the experimental data for Aedes albopictus population in Guangzhou, helps us predict a large range of releasing intensities for suppression success. In particular, we find that if the number of released infected males is no less than four times the number of mosquitoes in wild areas, then the mosquito density in the peak season can be reduced by 95%. We introduce an index to quantify the dependence of suppression efficiency on parameters. The invariance of some quantitative properties of the index values under various perturbations of the same parameter justifies the applicability of this index, and the robustness of our modeling approach. The index yields a ranking of the sensitivity of all parameters, among which the adult mortality has the highest sensitivity and is considerably more sensitive than the natural larvae mortality.}, } @article {pmid29234308, year = {2017}, author = {Chrostek, E and Pelz-Stelinski, K and Hurst, GDD and Hughes, GL}, title = {Horizontal Transmission of Intracellular Insect Symbionts via Plants.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {2237}, pmid = {29234308}, issn = {1664-302X}, support = {R21 AI124452/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; U01CK000512/ACL/ACL HHS/United States ; }, abstract = {Experimental evidence is accumulating that endosymbionts of phytophagous insects may transmit horizontally via plants. Intracellular symbionts known for manipulating insect reproduction and altering fitness (Rickettsia, Cardinium, Wolbachia, and bacterial parasite of the leafhopper Euscelidius variegatus) have been found to travel from infected insects into plants. Other insects, either of the same or different species can acquire the symbiont from the plant through feeding, and in some cases transfer it to their progeny. These reports prompt many questions regarding how intracellular insect symbionts are delivered to plants and how they affect them. Are symbionts passively transported along the insect-plant-insect path, or do they actively participate in the process? How widespread are these interactions? How does symbiont presence influence the plant? And what conditions are required for the new infection to establish in an insect? From an ecological, evolutionary, and applied perspective, this mode of horizontal transmission could have profound implications if occurring frequently enough or if new stable symbiont infections are established. Transmission of symbionts through plants likely represents an underappreciated means of infection, both in terms of symbiont epidemiology and the movement of symbionts to new host species.}, } @article {pmid29234159, year = {2018}, author = {Zug, R and Hammerstein, P}, title = {Evolution of reproductive parasites with direct fitness benefits.}, journal = {Heredity}, volume = {120}, number = {3}, pages = {266-281}, pmid = {29234159}, issn = {1365-2540}, mesh = {Animals ; Biological Evolution ; Female ; Fertility ; *Genetic Fitness ; Male ; *Models, Genetic ; Parasites/*physiology ; *Reproduction ; *Symbiosis ; }, abstract = {Maternally inherited symbionts such as Wolbachia have long been seen mainly as reproductive parasites, with deleterious effects on host fitness. It is becoming clear, however, that, frequently, these symbionts also have beneficial effects on host fitness, either along with reproductive parasitism or not. Using the examples of cytoplasmic incompatibility (CI) and male-killing (MK), we here analyze the effect of direct fitness benefits on the evolution of reproductive parasites. By means of a simple theoretical framework, we synthesize and extend earlier modeling approaches for CI and MK, which usually ignore fitness benefits. Moreover, our framework is not restricted to a particular mechanism underlying the fitness benefit (e.g., protection against pathogens). We derive invasion conditions and equilibrium frequencies for the different infection scenarios. Our results demonstrate the importance of a symbiont's "effective fecundity" (i.e., the product of the relative fecundity of an infected female and her transmission efficiency) for a symbiont's invasion success. In particular, we adopt the concept of effective fecundity to scenarios where CI and MK co-occur in one host population. We confirm that direct fitness benefits substantially facilitate the invasion and spread of infections (for example, by lowering or removing the invasion threshold) or even make invasion possible in the first place (for example, if reproductive parasitism is weak or absent). Finally, we discuss the role of direct fitness benefits in long-term evolutionary dynamics of reproductive phenotypes and highlight their potential to resolve genetic conflicts between maternally inherited symbionts and their hosts.}, } @article {pmid29216317, year = {2017}, author = {Fraser, JE and De Bruyne, JT and Iturbe-Ormaetxe, I and Stepnell, J and Burns, RL and Flores, HA and O'Neill, SL}, title = {Novel Wolbachia-transinfected Aedes aegypti mosquitoes possess diverse fitness and vector competence phenotypes.}, journal = {PLoS pathogens}, volume = {13}, number = {12}, pages = {e1006751}, pmid = {29216317}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/growth & development/*microbiology/physiology/virology ; Animals ; Communicable Disease Control/methods ; Culex/microbiology ; Dengue Virus/isolation & purification/physiology ; Drosophila melanogaster/microbiology ; Drosophila simulans/microbiology ; Female ; Fertility ; Infectious Disease Transmission, Vertical/*veterinary ; Male ; Mosquito Control/methods ; Mosquito Vectors/*microbiology/physiology/virology ; Organ Specificity ; Ovary/microbiology/physiology ; RNA, Viral/isolation & purification ; Salivary Glands/microbiology/physiology ; Sex Characteristics ; Species Specificity ; Survival Analysis ; Viral Tropism ; Wolbachia/*growth & development/isolation & purification ; }, abstract = {Wolbachia pipientis from Drosophila melanogaster (wMel) is an endosymbiotic bacterium that restricts transmission of human pathogenic flaviviruses and alphaviruses, including dengue, Zika, and chikungunya viruses, when introduced into the mosquito vector Aedes aegypti. To date, wMel-infected Ae. aegypti have been released in field trials in 5 countries to evaluate the effectiveness of this strategy for disease control. Despite the success in establishing wMel-infected mosquitoes in wild populations, and the well-characterized antiviral capabilities of wMel, transinfecting different or additional Wolbachia strains into Ae. aegypti may improve disease impact, and perhaps more importantly, could provide a strategy to account for the possible evolution of resistant arboviruses. Here, we report the successful transinfection of Ae. aegypti with the Wolbachia strains wMelCS (D. melanogaster), wRi (D. simulans) and wPip (Culex quinquefasciatus) and assess the effects on Ae. aegypti fitness, cytoplasmic incompatibility, tissue tropism and pathogen blocking in a laboratory setting. The results demonstrate that wMelCS provides a similar degree of protection against dengue virus as wMel following an infectious blood meal, and significantly reduces viral RNA levels beyond that of wMel following a direct challenge with infectious virus in mosquitoes, with no additional fitness cost to the host. The protection provided by wRi is markedly weaker than that of wMelCS, consistent with previous characterisations of these lines in Drosophila, while wPip was found to substantially reduce the fitness of Ae. aegypti. Thus, we determine wMelCS as a key candidate for further testing in field-relevant fitness tests and viremic blood feeding challenges in a clinical setting to determine if it may represent an alternative Wolbachia strain with more desirable attributes than wMel for future field testing.}, } @article {pmid29215202, year = {2018}, author = {Rock, DI and Smith, AH and Joffe, J and Albertus, A and Wong, N and O'Connor, M and Oliver, KM and Russell, JA}, title = {Context-dependent vertical transmission shapes strong endosymbiont community structure in the pea aphid, Acyrthosiphon pisum.}, journal = {Molecular ecology}, volume = {27}, number = {8}, pages = {2039-2056}, doi = {10.1111/mec.14449}, pmid = {29215202}, issn = {1365-294X}, mesh = {Animals ; Aphids/genetics/*microbiology ; Coinfection/genetics/*microbiology ; Coxiellaceae/genetics/pathogenicity ; *Ecology ; Host Specificity/genetics ; Microbiota/genetics ; Pisum sativum/parasitology ; Serratia/genetics/pathogenicity ; Symbiosis/*genetics ; }, abstract = {Animal-associated microbiomes are often comprised of structured, multispecies communities, with particular microbes showing trends of co-occurrence or exclusion. Such structure suggests variable community stability, or variable costs and benefits-possibilities with implications for symbiont-driven host adaptation. In this study, we performed systematic screening for maternally transmitted, facultative endosymbionts of the pea aphid, Acyrthosiphon pisum. Sampling across six locales, with up to 5 years of collection in each, netted significant and consistent trends of community structure. Co-infections between Serratia symbiotica and Rickettsiella viridis were more common than expected, while Rickettsia and X-type symbionts colonized aphids with Hamiltonella defensa more often than expected. Spiroplasma co-infected with other endosymbionts quite rarely, showing tendencies to colonize as a single species monoculture. Field estimates of maternal transmission rates help to explain our findings: while Serratia and Rickettsiella improved each other's transmission, Spiroplasma reduced transmission rates of co-infecting endosymbionts. In summary, our findings show that North American pea aphids harbour recurring combinations of facultative endosymbionts. Common symbiont partners play distinct roles in pea aphid biology, suggesting the creation of "generalist" aphids receiving symbiont-based defence against multiple ecological stressors. Multimodal selection, at the host level, may thus partially explain our results. But more conclusively, our findings show that within-host microbe interactions, and their resulting impacts on transmission rates, are an important determinant of community structure. Widespread distributions of heritable symbionts across plants and invertebrates hint at the far-reaching implications for these findings, and our work further shows the benefits of symbiosis research within a natural context.}, } @article {pmid29214346, year = {2018}, author = {Singhal, K and Mohanty, S}, title = {Comparative genomics reveals the presence of putative toxin-antitoxin system in Wolbachia genomes.}, journal = {Molecular genetics and genomics : MGG}, volume = {293}, number = {2}, pages = {525-540}, pmid = {29214346}, issn = {1617-4623}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/classification/genetics ; Drosophila/microbiology ; Drosophila melanogaster/microbiology ; Genes, Bacterial/genetics ; Genetic Variation ; Genome, Bacterial/*genetics ; Genomics/*methods ; Host-Pathogen Interactions ; Phylogeny ; Sequence Homology, Amino Acid ; Toxin-Antitoxin Systems/*genetics ; Wolbachia/*genetics/isolation & purification/physiology ; }, abstract = {Multiple toxin-antitoxin (TA) systems are housed in different locations within the bacterial genome and are known to be associated with various cellular processes and stress-related adaptation. In endosymbionts, although, the TA system has scarce occurrence but studies have highlighted its presence in enhancing host-symbiont interactions. Wolbachia, an obligate endosymbiont, has recently been proposed as a biocontrol agent which may be helpful in controlling vector-borne diseases. There are reports suggesting the role of TA system in inducing cytoplasmic incompatibility in case of Wolbachia, however, the underlying mechanism is still not known. The present study, therefore, aims at exploring the diversity of TA system in four novel (sourced from India) and three reference genomes (NCBI) of Wolbachia strains. Interestingly, we found several putative toxins and antitoxins of RelEB family of Type II TA system in these Wolbachia genomes. The results show wMel genome possessed more number of putative TA loci than wRi genome. In addition, searching through the other sequenced Wolbachia genomes in NCBI, a complete absence of TA system was observed in Wolbachia-infected nematodes. The sequence-wide analysis of all the putative RelEB proteins present amongst the Wolbachia endosymbiont and within the free-living bacterial genomes reveal strain-specific similarities and conserved sequences. However, large amount of sequence diversity was observed between Wolbachia and free-living bacteria. Understanding this sequence variation may help shed light on the differences between these two forms of bacteria and could also explain their niche preferences.}, } @article {pmid29194776, year = {2018}, author = {Kubiak, K and Sielawa, H and Chen, W and Dzika, E}, title = {Endosymbiosis and its significance in dermatology.}, journal = {Journal of the European Academy of Dermatology and Venereology : JEADV}, volume = {32}, number = {3}, pages = {347-354}, doi = {10.1111/jdv.14721}, pmid = {29194776}, issn = {1468-3083}, mesh = {Animals ; *Host-Pathogen Interactions ; Humans ; Insecta/microbiology ; Skin/*microbiology/*parasitology/virology ; Skin Diseases/*microbiology/*parasitology/virology ; *Symbiosis ; Wolbachia/physiology ; }, abstract = {Proposed at the beginning of the twentieth century to explain the origin of eukaryotic organelles from prokaryotes, endosymbiosis is now medically defined by various interaction patterns between microorganisms and their residing hosts, best exemplified by the bacterial endosymbiont Wolbachia identified in arthropods and filarial nematodes, which can influence normal development, reproduction, survival and transmission of the hosts. Based on the transmission modes, vertical or horizontal, and the function of the endosymbionts, the host-symbiont dependence can be divided into primary or secondary. In dermatology, the role of endosymbionts in skin ectoparasitosis has aroused great interests in the past years. Riesia pediculicola is a primary bacterial endosymbiont in body lice Pediculus humanus, and supplement their hosts with vitamin B, especially pantothenic acid. In cimicosis, the Gram-negative Wolbachia can synthesize biotin and riboflavin, which are crucial for the growth and reproduction of the bedbug Cimex lectularius. In human demodicosis and rosacea, further study is required to prove the pathogenic role of the Gram-negative bacteria Bacillus oleronius or the Gram-positive bacteria Bacillus cereus demonstrated in the Demodex mites. The high infection rate of adult female ticks Ixodes ricinus with the Gram-negative bacteria Midichloria mitochondrii present in the mitochondria in diverse ovarian cells, with the high seroprevalence rate in tick-exposed subjects, raises the possibility that this non-pathogenic endosymbiont may play a role in immune response and successful transmission of the tick-borne pathogen. The anaerobic protozoan Trichomonas vaginalis and bacteria Mycoplasma hominis are two obligate parasites in the urogenital epithelium, with partially overlapping symptoms. Intracellular localization of Mycoplasma hominis can avoid host immune response and penetration of antibiotics, while Trichomonas vaginalis infected with Mycoplasma hominis seems to have a higher cytopathic activity and amoeboid transformation rate. Further study on the biology and pathogenesis of different endosymbionts in dermatological parasitosis will help for the development of new treatment modalities.}, } @article {pmid31014588, year = {2017}, author = {González-Álvarez, VH and Fernández de Mera, IG and Cabezas-Cruz, A and de la Fuente, J and Ortega-Morales, AI and Almazán, C}, title = {Molecular survey of Rickettsial organisms in ectoparasites from a dog shelter in Northern Mexico.}, journal = {Veterinary parasitology, regional studies and reports}, volume = {10}, number = {}, pages = {143-148}, doi = {10.1016/j.vprsr.2017.10.005}, pmid = {31014588}, issn = {2405-9390}, mesh = {Animals ; Base Sequence ; DNA, Bacterial/*genetics ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Ectoparasitic Infestations/epidemiology/parasitology/*veterinary ; Housing, Animal ; Phthiraptera/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*genetics/isolation & purification ; Siphonaptera/microbiology ; Species Specificity ; Ticks/microbiology ; }, abstract = {The objective of this study was to screen and identify rickettsial organisms in ectoparasites collected from dogs in a shelter in Gomez Palacio, Durango, Mexico. One hundred dogs were inspected for ectoparasites. All the dogs were parasitized with Rhipicephalus sanguineus ticks, three with Heterodoxus spiniger lice and one with Ctenocephalides felis fleas. DNA was extracted from the ectoparasites found on each dog, and PCR with the primers for the Anaplasmataceae 16S rRNA and citrate synthase gltA genes were performed. Eight DNA samples obtained from ticks, three from lice and one from fleas were positive to 16S rRNA. Only one sample from C. felis and one from H. spiniger were positive to gltA. Sequence analysis of amplified products from C. felis showed identity to Rickettsia felis, Wolbachia pipientis, and Wolbachia spp., while a sequence from H. spiniger showed identity to Wolbachia spp. Herein we report the molecular detection of R. felis, W. pipientis, and Wolbachia spp. in C. felis and H. spiniger in northern Mexico. These results contribute to the knowledge of the microorganisms present in ectoparasites from dogs in Mexico.}, } @article {pmid29188381, year = {2017}, author = {Rahimi-Kaldeh, S and Ashouri, A and Bandani, A and Tomioka, K}, title = {The effect of Wolbachia on diapause, fecundity, and clock gene expression in Trichogramma brassicae (Hymenoptera: Trichogrammatidae).}, journal = {Development genes and evolution}, volume = {227}, number = {6}, pages = {401-410}, pmid = {29188381}, issn = {1432-041X}, mesh = {Animals ; CLOCK Proteins/*genetics ; *Diapause ; Female ; Gene Expression ; Insect Proteins/*genetics ; Photoperiod ; RNA, Messenger ; Wasps/*genetics/*physiology ; Wolbachia/*physiology ; }, abstract = {The short day lengths of late summer in moderate regions are used to induce diapause in various insects. Many studies have shown the maternal effect of photoperiod on diapause induction of Trichogramma wasps, but there is no study to show the relationship between photoperiodic regimes and clock genes in these useful biological control agents. Here, we investigated the role of photoperiods on diapause, fecundity, and clock gene expression (clk, cyc, cry2, per, and timeout) in asexual and sexual Trichogramma brassicae as a model insect to find any differences between two strains. Asexual strain was infected by Wolbachia, an endosymbiont bacterium. The diapause percentage was significantly higher under short days (8 h in sexual and 12 h in the asexual T. brassicae), although the diapause percentage of the sexual strain was significantly higher than the asexual one in all the photoperiods. The ANOVA revealed no significant changes between different photoperiods in the clock gene expression in the sexual strain but significant photoperiodic changes in clk, cyc, and timeout in the asexual strain. Our results showed that the mRNA levels of clock genes of asexual T. brassicae were significantly lower than those of sexual strain. The fecundity was significantly higher in the asexual strain. These results suggest that Wolbachia infection makes disturbance on the clock gene expression which consequently reduces the percentage of diapause but increases the fecundity in asexual T. brassicae.}, } @article {pmid29187976, year = {2017}, author = {Conner, WR and Blaxter, ML and Anfora, G and Ometto, L and Rota-Stabelli, O and Turelli, M}, title = {Genome comparisons indicate recent transfer of wRi-like Wolbachia between sister species Drosophila suzukii and D. subpulchrella.}, journal = {Ecology and evolution}, volume = {7}, number = {22}, pages = {9391-9404}, pmid = {29187976}, issn = {2045-7758}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, abstract = {Wolbachia endosymbionts may be acquired by horizontal transfer, by introgression through hybridization between closely related species, or by cladogenic retention during speciation. All three modes of acquisition have been demonstrated, but their relative frequency is largely unknown. Drosophila suzukii and its sister species D. subpulchrella harbor Wolbachia, denoted wSuz and wSpc, very closely related to wRi, identified in California populations of D. simulans. However, these variants differ in their induced phenotypes: wRi causes significant cytoplasmic incompatibility (CI) in D. simulans, but CI has not been detected in D. suzukii or D. subpulchrella. Our draft genomes of wSuz and wSpc contain full-length copies of 703 of the 734 single-copy genes found in wRi. Over these coding sequences, wSuz and wSpc differ by only 0.004% (i.e., 28 of 704,883 bp); they are sisters relative to wRi, from which each differs by 0.014%-0.015%. Using published data from D. melanogaster, Nasonia wasps and Nomada bees to calibrate relative rates of Wolbachia versus host nuclear divergence, we conclude that wSuz and wSpc are too similar-by at least a factor of 100-to be plausible candidates for cladogenic transmission. These three wRi-like Wolbachia, which differ in CI phenotype in their native hosts, have different numbers of orthologs of genes postulated to contribute to CI; and the CI loci differ at several nucleotides that may account for the CI difference. We discuss the general problem of distinguishing alternative modes of Wolbachia acquisition, focusing on the difficulties posed by limited knowledge of variation in absolute and relative rates of molecular evolution for host nuclear genomes, mitochondria, and Wolbachia.}, } @article {pmid29186405, year = {2018}, author = {Bleidorn, C and Gerth, M}, title = {A critical re-evaluation of multilocus sequence typing (MLST) efforts in Wolbachia.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {1}, pages = {}, doi = {10.1093/femsec/fix163}, pmid = {29186405}, issn = {1574-6941}, mesh = {Genetic Markers/genetics ; Genetic Variation/genetics ; Genome, Bacterial/*genetics ; Multilocus Sequence Typing/*methods ; Phylogeny ; Wolbachia/*genetics ; }, abstract = {Wolbachia (Alphaproteobacteria, Rickettsiales) is the most common, and arguably one of the most important inherited symbionts. Molecular differentiation of Wolbachia strains is routinely performed with a set of five multilocus sequence typing (MLST) markers. However, since its inception in 2006, the performance of MLST in Wolbachia strain typing has not been assessed objectively. Here, we evaluate the properties of Wolbachia MLST markers and compare it to 252 other single copy loci present in the genome of most Wolbachia strains. Specifically, we investigated how well MLST performs at strain differentiation, at reflecting genetic diversity of strains, and as phylogenetic marker. We find that MLST loci are outperformed by other loci at all tasks they are currently employed for, and thus that they do not reflect the properties of a Wolbachia strain very well. We argue that whole genome typing approaches should be used for Wolbachia typing in the future. Alternatively, if few loci approaches are necessary, we provide a characterisation of 252 single copy loci for a number a criteria, which may assist in designing specific typing systems or phylogenetic studies.}, } @article {pmid29183717, year = {2018}, author = {Dorigatti, I and McCormack, C and Nedjati-Gilani, G and Ferguson, NM}, title = {Using Wolbachia for Dengue Control: Insights from Modelling.}, journal = {Trends in parasitology}, volume = {34}, number = {2}, pages = {102-113}, pmid = {29183717}, issn = {1471-5007}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; U01 GM110721/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; *Biological Control Agents ; Dengue/*prevention & control/transmission ; Dengue Virus/*physiology ; Insect Vectors/microbiology ; *Models, Theoretical ; Wolbachia/*physiology ; }, abstract = {Dengue is the most common arboviral infection of humans, responsible for a substantial disease burden across the tropics. Traditional insecticide-based vector-control programmes have limited effectiveness, and the one licensed vaccine has a complex and imperfect efficacy profile. Strains of the bacterium Wolbachia, deliberately introduced into Aedes aegyptimosquitoes, have been shown to be able to spread to high frequencies in mosquito populations in release trials, and mosquitoes infected with these strains show markedly reduced vector competence. Thus, Wolbachia represents an exciting potential new form of biocontrol for arboviral diseases, including dengue. Here, we review how mathematical models give insight into the dynamics of the spread of Wolbachia, the potential impact of Wolbachia on dengue transmission, and we discuss the remaining challenges in evaluation and development.}, } @article {pmid29181449, year = {2017}, author = {Mann, E and Stouthamer, CM and Kelly, SE and Dzieciol, M and Hunter, MS and Schmitz-Esser, S}, title = {Transcriptome Sequencing Reveals Novel Candidate Genes for Cardinium hertigii-Caused Cytoplasmic Incompatibility and Host-Cell Interaction.}, journal = {mSystems}, volume = {2}, number = {6}, pages = {}, pmid = {29181449}, issn = {2379-5077}, abstract = {Cytoplasmic incompatibility (CI) is an intriguing, widespread, symbiont-induced reproductive failure that decreases offspring production of arthropods through crossing incompatibility of infected males with uninfected females or with females infected with a distinct symbiont genotype. For years, the molecular mechanism of CI remained unknown. Recent genomic, proteomic, biochemical, and cell biological studies have contributed to understanding of CI in the alphaproteobacterium Wolbachia and implicate genes associated with the WO prophage. Besides a recently discovered additional lineage of alphaproteobacterial symbionts only moderately related to Wolbachia, Cardinium (Bacteroidetes) is the only other symbiont known to cause CI, and genomic evidence suggests that it has very little homology with Wolbachia and evolved this phenotype independently. Here, we present the first transcriptomic study of the CI Cardinium strain cEper1, in its natural host, Encarsia suzannae, to detect important CI candidates and genes involved in the insect-Cardinium symbiosis. Highly expressed transcripts included genes involved in manipulating ubiquitination, apoptosis, and host DNA. Female-biased genes encoding ribosomal proteins suggest an increase in general translational activity of Cardinium in female wasps. The results confirm previous genomic analyses that indicated that Wolbachia and Cardinium utilize different genes to induce CI, and transcriptome patterns further highlight expression of some common pathways that these bacteria use to interact with the host and potentially cause this enigmatic and fundamental manipulation of host reproduction. IMPORTANCE The majority of insects carry maternally inherited intracellular bacteria that are important in their hosts' biology, ecology, and evolution. Some of these bacterial symbionts cause a reproductive failure known as cytoplasmic incompatibility (CI). In CI, the mating of symbiont-infected males and uninfected females produces few or no daughters. The CI symbiont then spreads and can have a significant impact on the insect host population. Cardinium, a bacterial endosymbiont of the parasitoid wasp Encarsia in the Bacteroidetes, is the only bacterial lineage known to cause CI outside the Alphaproteobacteria, where Wolbachia and another recently discovered CI symbiont reside. Here, we sought insight into the gene expression of a CI-inducing Cardinium strain in its natural host, Encarsia suzannae. Our study provides the first insights into the Cardinium transcriptome and provides support for the hypothesis that Wolbachia and Cardinium target similar host pathways with distinct and largely unrelated sets of genes.}, } @article {pmid29178391, year = {2018}, author = {Otten, C and Brilli, M and Vollmer, W and Viollier, PH and Salje, J}, title = {Peptidoglycan in obligate intracellular bacteria.}, journal = {Molecular microbiology}, volume = {107}, number = {2}, pages = {142-163}, pmid = {29178391}, issn = {1365-2958}, support = {/WT_/Wellcome Trust/United Kingdom ; MR/N501840/1/MRC_/Medical Research Council/United Kingdom ; 101824/Z/13/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Anaplasma marginale/classification/genetics/immunology/metabolism ; Animals ; *Bacteria/classification/genetics/immunology/metabolism ; Cell Wall/metabolism ; Chlamydia/classification/genetics/immunology/metabolism ; Cytoplasm/metabolism ; Genome, Bacterial/genetics ; *Host Microbial Interactions ; Humans ; Immunity, Innate/immunology ; Intracellular Space/*microbiology ; Orientia tsutsugamushi/classification/genetics/immunology/metabolism ; Peptidoglycan/chemistry/*genetics/*metabolism ; Phylogeny ; Wolbachia/classification/genetics/immunology/metabolism ; }, abstract = {Peptidoglycan is the predominant stress-bearing structure in the cell envelope of most bacteria, and also a potent stimulator of the eukaryotic immune system. Obligate intracellular bacteria replicate exclusively within the interior of living cells, an osmotically protected niche. Under these conditions peptidoglycan is not necessarily needed to maintain the integrity of the bacterial cell. Moreover, the presence of peptidoglycan puts bacteria at risk of detection and destruction by host peptidoglycan recognition factors and downstream effectors. This has resulted in a selective pressure and opportunity to reduce the levels of peptidoglycan. In this review we have analysed the occurrence of genes involved in peptidoglycan metabolism across the major obligate intracellular bacterial species. From this comparative analysis, we have identified a group of predicted 'peptidoglycan-intermediate' organisms that includes the Chlamydiae, Orientia tsutsugamushi, Wolbachia and Anaplasma marginale. This grouping is likely to reflect biological differences in their infection cycle compared with peptidoglycan-negative obligate intracellular bacteria such as Ehrlichia and Anaplasma phagocytophilum, as well as obligate intracellular bacteria with classical peptidoglycan such as Coxiella, Buchnera and members of the Rickettsia genus. The signature gene set of the peptidoglycan-intermediate group reveals insights into minimal enzymatic requirements for building a peptidoglycan-like sacculus and/or division septum.}, } @article {pmid29169377, year = {2017}, author = {Pagès, N and Muñoz-Muñoz, F and Verdún, M and Pujol, N and Talavera, S}, title = {First detection of Wolbachia-infected Culicoides (Diptera: Ceratopogonidae) in Europe: Wolbachia and Cardinium infection across Culicoides communities revealed in Spain.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {582}, pmid = {29169377}, issn = {1756-3305}, support = {FAU2008-0019//INIA/ ; }, mesh = {Animals ; Bacterial Infections/*epidemiology/microbiology ; Bacteroidetes/genetics/*isolation & purification ; Bluetongue/transmission ; Ceratopogonidae/classification/*microbiology ; Disease Vectors ; Europe/epidemiology ; Gram-Negative Bacterial Infections/epidemiology/microbiology ; Insect Control ; Insect Vectors/*microbiology ; Prevalence ; Spain/epidemiology ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) transmit pathogens that cause important diseases. No effective technique has been found to properly control either Culicoides spp. abundance or their likelihood to transmit pathogens. Endosymbionts, particularly Wolbachia, represent powerful alternatives to control arthropods of health interest. In arthropods, Wolbachia can reduce vector fitness and vector's pathogen transmission capacity, thus being a potential target for population reduction and replacement strategies.

RESULTS: The presence of Wolbachia and Cardinium endosymbionts was screened in Spanish Culicoides spp. populations at livestock premises and natural habitats. The first detection of Wolbachia-infected Culicoides spp. in Europe is reported. The putative Palaearctic vectors for bluetongue and Schmallenberg diseases, C. imicola, C. obsoletus (s.s.) and C. pulicaris (s.l.), were infected with Wolbachia. Four genetic clusters of closely-related Wolbachia strains from A and B supergroups were detected infecting Culicoides. Cardinium strain of the C-group was detected in C. obsoletus (s.l.). Both endosymbionts, Wolbachia and Cardinium, were detected in Culicoides species of minor epidemiological relevance as well. Higher prevalence of Wolbachia infection was detected in natural habitats, while livestock premises lead to higher prevalence of Cardinium. Significant differences in the prevalence of Wolbachia, but not Cardinium, were also detected between some Culicoides species and between locations.

CONCLUSIONS: The presence of Wolbachia and Cardinium endosymbionts in Culicoides is expected to trigger new research towards the control of Culicoides-transmitted diseases. The results of the present study could have an impact beyond the Culicoides arena because successful Wolbachia transfection is possible even across genus and species barriers.}, } @article {pmid29165845, year = {2018}, author = {Audsley, MD and Seleznev, A and Joubert, DA and Woolfit, M and O'Neill, SL and McGraw, EA}, title = {Wolbachia infection alters the relative abundance of resident bacteria in adult Aedes aegypti mosquitoes, but not larvae.}, journal = {Molecular ecology}, volume = {27}, number = {1}, pages = {297-309}, doi = {10.1111/mec.14436}, pmid = {29165845}, issn = {1365-294X}, mesh = {Aedes/*microbiology ; Animals ; Australia ; Biodiversity ; Female ; Genes, Bacterial ; High-Throughput Nucleotide Sequencing ; Larva/microbiology ; Microbiota/genetics ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Wolbachia/classification/*physiology ; }, abstract = {Insect-symbiont interactions are known to play key roles in host functions and fitness. The common insect endosymbiont Wolbachia can reduce the ability of several human pathogens, including arboviruses and the malaria parasite, to replicate in insect hosts. Wolbachia does not naturally infect Aedes aegypti, the primary vector of dengue virus, but transinfected Ae. aegypti have antidengue virus properties and are currently being trialled as a dengue biocontrol strategy. Here, we assess the impact of Wolbachia infection of Ae. aegypti on the microbiome of wild mosquito populations (adults and larvae) collected from release sites in Cairns, Australia, by profiling the 16S rRNA gene using next-generation sequencing. Our data indicate that Wolbachia reduces the relative abundance of a large proportion of bacterial taxa in Ae. aegypti adults, that is in accordance with the known pathogen-blocking effects of Wolbachia on a variety of bacteria and viruses. In adults, several of the most abundant bacterial genera were found to undergo significant shifts in relative abundance. However, the genera showing the greatest changes in relative abundance in Wolbachia-infected adults represented a low proportion of the total microbiome. In addition, there was little effect of Wolbachia infection on the relative abundance of bacterial taxa in larvae, or on species diversity (accounting for species richness and evenness together) detected in adults or larvae. These results offer insight into the effects of Wolbachia on the Ae. aegypti microbiome in a native setting, an important consideration for field releases of Wolbachia into the population.}, } @article {pmid29161848, year = {2018}, author = {Zheng, B and Guo, W and Hu, L and Huang, M and Yu, J}, title = {Complex wolbachia infection dynamics in mosquitoes with imperfect maternal transmission.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {15}, number = {2}, pages = {523-541}, doi = {10.3934/mbe.2018024}, pmid = {29161848}, issn = {1551-0018}, mesh = {Aedes/*microbiology ; Algorithms ; Animals ; Australia ; Dengue/*transmission ; Female ; Humans ; Male ; *Models, Theoretical ; Mosquito Vectors/*microbiology ; Oviposition ; *Pest Control, Biological ; Vietnam ; *Wolbachia ; }, abstract = {Dengue, malaria, and Zika are dangerous diseases primarily transmitted by Aedes aegypti, Aedes albopictus, and Anopheles stephensi. In the last few years, a new disease control method, besides pesticide spraying to kill mosquitoes, has been developed by releasing mosquitoes carrying bacterium Wolbachia into the natural areas to infect the wild population of mosquitoes and block disease transmission. The bacterium is transmitted by infected mothers and the maternal transmission was assumed to be perfect in virtually all previous models. However, recent experiments on Aedes aegypti and Anopheles stephensi showed that the transmission can be imperfect. In this work, we develop a model to describe how the imperfect maternal transmission affects the dynamics of Wolbachia spread. We establish two useful identities and employ them to find sufficient and necessary conditions under which the system exhibits monomorphic, bistable, and polymorphic dynamics. These analytical results may help find a plausible explanation for the recent observation that the Wolbachia strain ωMelPop failed to establish in the natural populations in Australia and Vietnam.}, } @article {pmid29160361, year = {2018}, author = {Almeida, RP and Stouthamer, R}, title = {Phylogeny of the Trichogramma endosymbiont Wolbachia, an alpha-proteobacteria (Rickettsiae).}, journal = {Brazilian journal of biology = Revista brasleira de biologia}, volume = {78}, number = {3}, pages = {421-428}, doi = {10.1590/1519-6984.166671}, pmid = {29160361}, issn = {1678-4375}, mesh = {Alphaproteobacteria/*metabolism ; Animals ; Bacterial Outer Membrane Proteins/*metabolism ; DNA Primers/*genetics ; Female ; Genes, Bacterial/*genetics ; Phylogeny ; Reproduction ; Species Specificity ; Symbiosis ; Wasps/genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia (Hertig) endosymbionts are extensively studied in a wide range of organisms and are known to be transmitted through the egg cytoplasm to the offsping. Wolbachia may cause several types of reproductive modifications in arthropods. In Trichogramma species, parthenogenesis-inducing Wolbachia bacteria allow females wasps to produce daughters from unfertilized eggs and these bacteria are present in at least 9% of all Trichogramma species. Phylogenetic studies have led to the subdivision of the Wolbachia clade in five supergroups (A, B, C, D and E) and Wolbachia from Trichogramma belong to supergroup B. Here, using the wsp gene, four groups of Wolbachia that infect Trichogramma species were distinguished and the addition of a new group "Ato" was suggested due to the addition of Wolbachia from Trichogramma atopovirilia (Oatman and Platner). Specific primers were designed and tested for the "Ato" group. Seventy-five percent of all evaluated Wolbachia strains from Trichogramma fell within "Sib" group.}, } @article {pmid29157676, year = {2018}, author = {Asad, S and Parry, R and Asgari, S}, title = {Upregulation of Aedes aegypti Vago1 by Wolbachia and its effect on dengue virus replication.}, journal = {Insect biochemistry and molecular biology}, volume = {92}, number = {}, pages = {45-52}, doi = {10.1016/j.ibmb.2017.11.008}, pmid = {29157676}, issn = {1879-0240}, mesh = {Aedes/*genetics/metabolism/*microbiology ; Animals ; Cell Line ; Dengue Virus/*physiology ; Female ; Gene Knockdown Techniques ; Insect Proteins/*genetics/metabolism ; *Up-Regulation ; *Virus Replication ; Wolbachia/*physiology ; }, abstract = {Dengue infection along with its related disease conditions poses a significant threat to human health. The pathogen responsible for this infection is dengue virus (DENV) which is primarily transmitted to humans through the bites of Aedes aegypti mosquitoes. Unavailability of a potent vaccine has recently sparked renewed research endeavours aimed at vector control. To date, Wolbachia as an endosymbiotic bacterium has shown promise as a novel biocontrol agent to restrict DENV replication in the vector, although the underlying antiviral mechanism remains elusive. Recent studies have demonstrated the potential role of Vago as a novel secretory protein involved in cross-talk between the innate immune pathways in Culex quinquefasciatus mosquitoes to restrict West Nile virus replication. In this study, we have identified two homologs of the Vago protein in Ae. aegypti and looked into their modulation in the case of Wolbachia wMelPop strain infection. Furthermore, we have investigated the role of AeVago1, that is highly induced by Wolbachia, in the context of Wolbachia-mosquito-DENV interactions. Knockdown studies of the AeVago1 gene in Wolbachia-infected cells led to significant increases in DENV replication, with no effect on Wolbachia density. Our results suggest that the Wolbachia-induced AeVago1 in Ae. aegypti may function as a host factor to suppress DENV replication in the mosquito.}, } @article {pmid29150322, year = {2018}, author = {Van Oosten, AR and Duron, O and Heylen, DJA}, title = {Sex ratios of the tick Ixodes arboricola are strongly female-biased, but there are no indications of sex-distorting bacteria.}, journal = {Ticks and tick-borne diseases}, volume = {9}, number = {2}, pages = {307-313}, doi = {10.1016/j.ttbdis.2017.11.004}, pmid = {29150322}, issn = {1877-9603}, mesh = {Animals ; Bacterial Physiological Phenomena ; Belgium ; Coxiellaceae/*physiology ; Ixodes/*microbiology/*physiology ; Rickettsia/*physiology ; *Sex Ratio ; Songbirds/parasitology ; Spiroplasma/*physiology ; }, abstract = {Studies on sex ratio are of fundamental importance for understanding the biology of populations and biological control of pests and pathogens. In most Ixodes tick species, only females feed in the adult stage and, hence, contribute to pathogen transmission. The tree-hole tick Ixodes arboricola infests cavity-nesting birds and has limited dispersal possibilities. It plays an important role in the maintenance of zoonotic disease cycles. Here, we quantified the sex ratio of 718 adult I. arboricola ticks obtained from a laboratory stock at nine distinct periods (cohorts) from 2008 to 2015. In addition, we screened 93 specimens, collected from four study sites in 2011 and 2012, for the presence of six maternally inherited bacterial parasites known to manipulate arthropod sex ratios. We found significantly female-biased sex ratios in seven out of nine cohorts. There were no infections with members of the Wolbachia, Arsenophonus or Cardinium bacterial genera, whereas 96.8% of the screened ticks were infected with Rickettsia vini, 22.6% with Rickettsiella sp., and 14.0% with Spiroplasma ixodetis. Male and female I. arboricola were found equally infected. Our results suggest skewed sex ratios in I. arboricola are not caused by these bacterial infections, although there may be other, untested candidates driving sex ratios. Alternatively, female-biased sex ratios may be an adaptation in females to high local densities and low dispersal, where the production of daughters has a selective advantage because a few sons can fertilise all daughters.}, } @article {pmid29150304, year = {2017}, author = {Lichten, E}, title = {Are the estrogenic hormonal effects of environmental toxins affecting small intestinal bacterial and microfilaria overgrowth?.}, journal = {Medical hypotheses}, volume = {109}, number = {}, pages = {90-94}, doi = {10.1016/j.mehy.2017.09.022}, pmid = {29150304}, issn = {1532-2777}, mesh = {Adult ; Animals ; Anti-Bacterial Agents/therapeutic use ; Bacterial Infections/pathology ; Colitis, Ulcerative/pathology ; Crohn Disease/pathology ; Endocrine Disruptors/*adverse effects ; Environmental Exposure/*adverse effects ; Estrogens/adverse effects ; Female ; Humans ; Intestine, Small/*drug effects/*microbiology/*parasitology ; Male ; Microfilariae/growth & development ; Models, Theoretical ; Nematoda/*growth & development ; Rifamycins/therapeutic use ; Rifaximin ; Testosterone Congeners/pharmacology ; Treatment Outcome ; Wolbachia/growth & development ; }, abstract = {The important role of microfilaria (worms) in human and animal disease remains an area of key disagreement between the naturopathic and allopathic physicians. While microfilaria infections are rampart in undeveloped countries, they rarely rise to identification as a cause of disease in Western countries. New research studies in the diagnosis and treatment of SIBO (Small Intestinal Bacterial Overgrowth) and (IBD) Inflammatory Bowel Diseases of ulcerative colitis, Crohn's Disease and microcytic colitis may make both sides equally correct. A study of rifaximin failures in SIBO positive individuals finds biomarkers of decreased Free Androgen Index (FAI), high incidence of autoimmune disease and elevated Sex Hormone Binding Globulin (SHBG). The author hypothesizes that the underlying pathophysiology is increased exposure to Endocrine Disrupting Chemicals (EDCs) which hormonally act as xeno-estrogens. These xeno-estrogens increase the host production of SHBG, reduce pituitary stimulation of androgen product and result in a shift to estrogen dominance. Estrogen dominance is associated with autoimmune diseases and catabolic states. Treatment with a mixture of anabolic steroids that raises the FAI and lowers SHBG results in dramatic improvement in the signs and symptoms and recovery of the vast percentage of severe SIBO sufferers the author has treated. Similar results have been seen in severe pre-surgical cases of IBD whom fail all pharmaceutical interventions. Based on the recent recognition of the biological importance of Wolbachia in the occurrence of major diseases in the underdeveloped countries such as onchocerciasis, and the sexual nature of Wolbachia's role in helminths reproduction, the author hypothesizes that the EDCs are shifting the host's hormonal milieu in a more estrogenic direction and increasing reproduction of helminths changing the gastrointestinal microbiota. Present allopathic treatment of onchocerciasis utilizes albendazole and avermectin as therapy against the microfilaria larvae and doxycycline as bactericidal for Wolbachia. The allopathic treatments are unacceptable for pregnancy and children. Both naturopathic and allopathic treatments share a common focus on the suppression of the underlying bacterium Wolbachia infestation. The author hypothesizes that treatment of these two very different gastrointestinal diseases involves first establishing a normal, anabolic hormonal milieu and concurrently controlling an underlying yet unrecognized microfilaria overgrowth through naturopathic and allopathic treatments prescribed to the host. A case report of one such critically ill individual is noted. A thorough case controlled observation of symptoms matched with biological culture colony count and concentration of microfilaria in disease before and after the aforementioned anabolic treatment may answer the hypothesis.}, } @article {pmid29146940, year = {2017}, author = {Dutra, HLC and Rodrigues, SL and Mansur, SB and de Oliveira, SP and Caragata, EP and Moreira, LA}, title = {Development and physiological effects of an artificial diet for Wolbachia-infected Aedes aegypti.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {15687}, pmid = {29146940}, issn = {2045-2322}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Cattle ; Cytoplasm/metabolism ; *Diet ; Erythrocytes/metabolism/parasitology ; Feeding Behavior ; Female ; Fertility ; Humans ; Longevity ; Male ; Wolbachia/*physiology ; Zika Virus/physiology ; Zika Virus Infection/virology ; }, abstract = {The endosymbiotic bacterium Wolbachia spreads rapidly through populations of Aedes aegypti mosquitoes, and strongly inhibits infection with key human pathogens including the dengue and Zika viruses. Mosquito control programs aimed at limiting transmission of these viruses are ongoing in multiple countries, yet there is a dearth of mass rearing infrastructure specific to Wolbachia-infected mosquitoes. One example is the lack of a blood meal substitute, which accounts for the Wolbachia-specific physiological changes in infected mosquitoes, that allows the bacterium to spread, and block viral infections. To that end, we have developed a blood meal substitute specifically for mosquitoes infected with the wMel Wolbachia strain. This diet, ADM, contains milk protein, and infant formula, dissolved in a mixture of bovine red blood cells and Aedes physiological saline, with ATP as a phagostimulant. Feeding with ADM leads to high levels of viable egg production, but also does not affect key Wolbachia parameters including, bacterial density, cytoplasmic incompatibility, or resistance to infection with Zika virus. ADM represents an effective substitute for human blood, which could potentially be used for the mass rearing of wMel-infected A. aegypti, and could easily be optimized in the future to improve performance.}, } @article {pmid29143669, year = {2017}, author = {Papich, MG}, title = {Considerations for using minocycline vs doxycycline for treatment of canine heartworm disease.}, journal = {Parasites & vectors}, volume = {10}, number = {Suppl 2}, pages = {493}, pmid = {29143669}, issn = {1756-3305}, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage/pharmacokinetics ; Dirofilaria immitis/*microbiology/physiology ; Dirofilariasis/*drug therapy/parasitology ; Dog Diseases/*drug therapy/parasitology ; Dogs ; Doxycycline/*administration & dosage/pharmacokinetics ; Mice ; Minocycline/*administration & dosage/pharmacokinetics ; Wolbachia/*drug effects/physiology ; }, abstract = {BACKGROUND: Doxycycline has been considered the first drug of choice for treating Wolbachia, a member of the Rickettsiaceae, which has a symbiotic relationship with filarial worms, including heartworms. Wolbachia, is susceptible to tetracyclines, which have been used as adjunctive treatments for heartworm disease. Treatment with doxycycline reduces Wolbachia numbers in all stages of heartworms and improves outcomes and decreased microfilaremia in dogs treated for heartworm disease. The American Heartworm Society recommends treatment with doxycycline in dogs diagnosed with heartworm disease at a dose of 10 mg/kg twice daily for 28 days. If doxycycline is not available, minocycline can be considered as a substitute. However, minocycline has not undergone an evaluation in dogs with heartworm disease, nor has an effective dose been established. Minocycline is an attractive option because of the higher cost of doxycycline and new pharmacokinetic information for dogs that provides guidance for appropriate dosage regimens to achieve pharmacokinetic-pharmacodynamic (PK-PD) targets.

RESULTS: Published reports from the Anti-Wolbachia Consortium (A-WOL) indicate superior in vitro activity of minocycline over doxycycline. Studies performed in mouse models to measure anti-Wolbachia activity showed that minocycline was 1.7 times more effective than doxycycline, despite a 3-fold lower pharmacokinetic exposure. To achieve the same exposure as achieved in the mouse infection model, a pharmacokinetic-pharmacodynamic (PK-PD) analysis was conducted to determine optimal dosages for dogs. The analysis showed that an oral minocycline dose of 3.75 to 5 mg/kg administered twice daily would attain similar targets as observed in mice and predicted for human infections.

CONCLUSIONS: There are potentially several advantages for use of minocycline in animals. It is well absorbed from oral administration, it has less protein binding than doxycycline (65% vs 92%) allowing for better distribution into tissue, and it is approximately two times more lipophilic than doxycycline, which may result in better intracellular penetration. More work is needed to document efficacy of minocycline for treating canine heartworm disease.}, } @article {pmid29143657, year = {2017}, author = {Nelson, CT and Myrick, ES and Nelson, TA}, title = {Clinical benefits of incorporating doxycycline into a canine heartworm treatment protocol.}, journal = {Parasites & vectors}, volume = {10}, number = {Suppl 2}, pages = {515}, pmid = {29143657}, issn = {1756-3305}, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage ; Clinical Protocols ; Dirofilaria immitis/drug effects/physiology ; Dirofilariasis/*drug therapy/parasitology ; Dog Diseases/*drug therapy/parasitology ; Dogs ; Female ; Filaricides/*administration & dosage ; Ivermectin/administration & dosage ; Male ; Wolbachia/*drug effects/physiology ; }, abstract = {BACKGROUND: The objective of heartworm treatment is to improve the clinical condition of the patient and to eliminate pre-cardiac, juvenile, and adult worm stages with minimal complications. Pulmonary thromboembolisms are an inevitable consequence of worm death and can result in severe pulmonary reactions and even death of the patient. To minimize these reactions, various treatment protocols involving melarsomine, the only adulticidal drug approved by the US Food and Drug Administrations (FDA), in conjunction with macrocyclic lactone heartworm preventives and glucocorticosteroids have been advocated. The discovery of the bacterial endosymbiont Wolbachia in Dirofilaria immitis has led to several experimental studies examining the effects of administering doxycycline to reduce or eliminate Wolbachia organism. These studies have shown a decrease in gross and microscopic pathology of pulmonary parenchyma in experimental heartworm infections pretreated with doxycycline before melarsomine administration.

METHODS: Electronic medical records from a large veterinary practice in northeast Alabama were searched to identify dogs treated for heartworms with melarsomine from January 2005 through December 2012. The search was refined further to select for dogs that met the following criteria: 1) received two or three doses of ivermectin heartworm preventive prior to melarsomine injections, 2) received one injection of melarsomine followed by two injections 4 to 8 weeks later, and 3) were treated with prednisone following melarsomine injections. The dogs were then divided into those that also were treated with doxycycline 10 mg/kg BID for 4 weeks (Group A, n = 47) and those that did not receive doxycycline (Group B, n = 47). The medical notes of all 94 cases were then reviewed for comments concerning coughing, dyspnea, or hemoptysis in the history, physical exam template, or from telephone conversations with clients the week following each visit. Any dog that died within one year of treatment from either cardiovascular or pulmonary problems was noted.

RESULTS: Dogs from Group A receiving doxycycline had fewer respiratory complications (6.52%) and heartworm disease-related deaths (0%) than Group B (19.14% and 4.25%, respectively).

CONCLUSIONS: Although there are not enough cases to indicate statistical significance, the results strongly suggest that including doxycycline into canine heartworm treatment protocols decreases post-treatment complications and mortality in naturally infected clinical cases.}, } @article {pmid29141766, year = {2018}, author = {Sarwar, MS and Jahan, N and Shahbaz, F}, title = {Molecular Detection and Characterization of Wolbachia pipientis from Culex quinquefasciatus Collected from Lahore, Pakistan.}, journal = {The American journal of tropical medicine and hygiene}, volume = {98}, number = {1}, pages = {154-161}, pmid = {29141766}, issn = {1476-1645}, mesh = {Animals ; Culex/*microbiology ; DNA, Bacterial/genetics ; Electrophoresis, Agar Gel ; Female ; Male ; Ovary/microbiology ; Pakistan ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {The gram-negative, pleomorphic endosymbiont Wolbachia is known to infect a large number of insects and other arthropods naturally. This bacterium modifies the host biology, mainly causing reproductive alterations including feminization, death of male, parthenogenesis, and importantly cytoplasmic incompatibility. Wolbachia-induced cytoplasmic incompatibility results in nonviable offspring and vector population suppression. In addition, this bacterium rapidly spreads and propagates within the host population. This study is the first report on Wolbachia detection and characterization from Culex quinquefasciatus collected from Lahore, Pakistan. For this purpose, mosquito adults were collected from different localities of Lahore and identified at the species level. A total of 145 pairs of ovaries were individually subjected to DNA isolation, and polymerase chain reaction amplification of three (wsp, 16S rRNA, and ftsZ) genes were investigated. In all, 128 females were found positive, representing 82.3% infection rate. The phylogenetic analysis indicated that the detected endosymbiont had 100% homology with Wolbachia pipientis wPip strain and supergroup B. The detection of the local strain of Wolbachia (wPip) will be useful in investigating its potential for the control of dengue vector (Aedes aegypti) and reducing dengue transmission in Pakistan.}, } @article {pmid29132425, year = {2017}, author = {Zhang, D and Zhang, M and Wu, Y and Gilles, JRL and Yamada, H and Wu, Z and Xi, Z and Zheng, X}, title = {Establishment of a medium-scale mosquito facility: optimization of the larval mass-rearing unit for Aedes albopictus (Diptera: Culicidae).}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {569}, pmid = {29132425}, issn = {1756-3305}, support = {19115//the Coordinated Research Project D44002 from IAEA/ ; R01AI080597//the National Institutes of Health/National Institute of Allergy and Infectious Disease/ ; No. 311030//the Key Project of Chinese Ministry of Education/ ; No. 2011S009//the Guangdong Innovative Research Team Program/ ; No. 2016-2414//the Scientific and Technological Leading Talents of Guangzhou Development District/ ; No. 2016YFC1200500//National Key R & D Program of China/ ; R01 AI080597/AI/NIAID NIH HHS/United States ; 18816//the Coordinated Research Project D42016 from IAEA/ ; }, mesh = {Aedes/genetics/growth & development/microbiology/*physiology ; Animals ; Female ; Larva/growth & development/physiology ; Life Cycle Stages ; Male ; Pupa/growth & development/physiology ; Reproduction ; Wolbachia/physiology ; }, abstract = {BACKGROUND: Standardized larval rearing units for mosquito production are essential for the establishment of a mass-rearing facility. Two larval rearing units, developed respectively by the Guangzhou Wolbaki Biotech Co. Ltd. (Wolbaki) and Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture (FAO/IAEA-IPCL), are tested to assess their potential uses to mass-rear the larval stages of Aedes albopictus in support of the establishment of a medium-scale mosquito facility for the application of mosquito genetic control strategies.

METHODS: The triple Wolbachia-infected Ae. albopictus strain (HC strain) was used in this study. The effects of larval densities of two larval rearing trays (corresponding to 2.4, 3.0 and 3.6 larvae/cm[2]) and tray size/position (top, middle and bottom layers) on the pupae production and larval survival were assessed when trays were stacked within the larval rearing units. The male pupae production, female pupae contamination after sex separation, and male mating competitiveness were also studied by using both larval rearing units in their entirety.

RESULTS: The optimal larval rearing density for Wolbaki-tray (Wol-tray) was 6,600 larvae (equal to 3.0 larvae/cm[2]) and 18,000 larvae (3.6 larvae/cm[2]) for the FAO/IAEA-IPCL tray (IAEA-tray). No significant difference in pupae production was observed when trays were stacked within top, middle or bottom layers for both units. At thirty-four hours after the first pupation, the average male pupae production was (0.89 × 10[5]) for the Wol-unit and (3.16 × 10[5]) for the IAEA-unit. No significant difference was observed in female pupae contamination between these two units. The HC males showed equal male mating competitiveness to wild type males for mating with wild type females in large cages, regardless of whether they were reared in the Wol-unit or IAEA-unit.

CONCLUSIONS: The current study has indicated that both the Wol-unit and IAEA-unit are suitable for larvae mass-rearing for Ae. albopictus. However, the IAEA-unit, with higher male production and less space required compared to the Wol-unit, is recommended to be used in support of the establishment of a medium-sized mosquito facility.}, } @article {pmid29125253, year = {2017}, author = {Tmimi, FZ and Bkhache, M and Mounaji, K and Failloux, AB and Sarih, M}, title = {First report of the endobacteria Wolbachia in natural populations of Culex pipiens in Morocco.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {42}, number = {2}, pages = {349-351}, doi = {10.1111/jvec.12275}, pmid = {29125253}, issn = {1948-7134}, mesh = {Animals ; Culex/*microbiology ; Morocco ; Wolbachia/*physiology ; }, } @article {pmid29122518, year = {2017}, author = {Frentiu, FD}, title = {Lipids and Pathogen Blocking by Wolbachia.}, journal = {Trends in parasitology}, volume = {33}, number = {12}, pages = {916-917}, doi = {10.1016/j.pt.2017.10.007}, pmid = {29122518}, issn = {1471-5007}, mesh = {Aedes/*microbiology/*virology ; Animals ; Dengue/microbiology/prevention & control/transmission/virology ; Dengue Virus/physiology ; Disease Transmission, Infectious/*prevention & control ; Lipids/*physiology ; Microbial Interactions/*physiology ; Mosquito Vectors/microbiology/virology ; Wolbachia/*metabolism/*virology ; Zika Virus/physiology ; Zika Virus Infection/microbiology/prevention & control/transmission/virology ; }, abstract = {Mosquito-borne viruses are major human pathogens. Introducing Wolbachia into mosquitoes could reduce disease burdens because these bacteria block virus transmission. How Wolbachia does this is unclear, but new data show that modulation of host-cell lipids is critical.}, } @article {pmid29116011, year = {2017}, author = {Tsai, CH and Chen, TH and Lin, C and Shu, PY and Su, CL and Teng, HJ}, title = {The impact of temperature and Wolbachia infection on vector competence of potential dengue vectors Aedes aegypti and Aedes albopictus in the transmission of dengue virus serotype 1 in southern Taiwan.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {551}, pmid = {29116011}, issn = {1756-3305}, mesh = {Aedes/anatomy & histology/*microbiology/physiology/*virology ; Animals ; Dengue/epidemiology/transmission/virology ; Dengue Virus/*physiology ; Female ; Mosquito Vectors/*microbiology/physiology/*virology ; Salivary Glands/virology ; Taiwan/epidemiology ; Temperature ; Thorax ; Viral Load ; Virus Replication ; Wolbachia/isolation & purification/*physiology ; }, abstract = {BACKGROUND: We evaluated the impact of temperature and Wolbachia infection on vector competence of the local Aedes aegypti and Ae. albopictus populations of southern Taiwan in the laboratory.

RESULTS: After oral infection with dengue serotype 1 virus (DENV-1), female mosquitoes were incubated at temperatures of 10, 16, 22, 28 and 34 °C. Subsequently, salivary gland, head, and thorax-abdomen samples were analyzed for their virus titer at 0, 5, 10, 15, 20, 25 and 30 days post-infection (dpi) by real-time RT-PCR. The results showed that Ae. aegypti survived significantly longer and that dengue viral genome levels in the thorax-abdomen (10[3.25 ± 0.53]-10[4.09 ± 0.71] PFU equivalents/ml) and salivary gland samples (10[2.67 ± 0.33]-10[3.89 ± 0.58] PFU equivalents/ml) were significantly higher at high temperature (28-34 °C). The survival of Ae. albopictus was significantly better at 16 or 28 °C, but the virus titers from thorax-abdomen (10[0.70]-10[2.39 ± 1.31] PFU equivalents/ml) and salivary gland samples (10[0.12 ± 0.05]-10[1.51 ± 0.31] PFU equivalents/ml) were significantly higher at 22-28 °C. Within viable temperature ranges, the viruses were detectable after 10 dpi in salivary glands and head tissues in Ae. aegypti and after 5-10 dpi in Ae. albopictus. Vector competence was measured in Ae. albopictus with and without Wolbachia at 28 °C. Wolbachia-infected mosquitoes survived significantly better and carried lower virus titers than Wolbachia-free mosquitoes. Wolbachia coinfections (92.8-97.2%) with wAlbA and wAlbB strains were commonly found in a wild population of Ae. albopictus.

CONCLUSIONS: In southern Taiwan, Ae. aegypti is the main vector of dengue and Ae. albopictus has a non-significant role in the transmission of dengue virus due to the high prevalence of Wolbachia infection in the local mosquito population of southern Taiwan.}, } @article {pmid29114401, year = {2017}, author = {Robinson, KM and Hawkins, AS and Santana-Cruz, I and Adkins, RS and Shetty, AC and Nagaraj, S and Sadzewicz, L and Tallon, LJ and Rasko, DA and Fraser, CM and Mahurkar, A and Silva, JC and Dunning Hotopp, JC}, title = {Aligner optimization increases accuracy and decreases compute times in multi-species sequence data.}, journal = {Microbial genomics}, volume = {3}, number = {9}, pages = {e000122}, pmid = {29114401}, issn = {2057-5858}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/genetics ; Chromosome Mapping ; Data Accuracy ; Databases, Genetic ; Datasets as Topic ; Genome, Human ; High-Throughput Nucleotide Sequencing ; Humans ; Metagenomics ; Plasmodium falciparum/genetics ; Sequence Alignment/*methods ; *Sequence Analysis, DNA ; *Software ; Time Factors ; Wolbachia/genetics ; }, abstract = {As sequencing technologies have evolved, the tools to analyze these sequences have made similar advances. However, for multi-species samples, we observed important and adverse differences in alignment specificity and computation time for bwa- mem (Burrows-Wheeler aligner-maximum exact matches) relative to bwa-aln. Therefore, we sought to optimize bwa-mem for alignment of data from multi-species samples in order to reduce alignment time and increase the specificity of alignments. In the multi-species cases examined, there was one majority member (i.e. Plasmodium falciparum or Brugia malayi) and one minority member (i.e. human or the Wolbachia endosymbiont wBm) of the sequence data. Increasing bwa-mem seed length from the default value reduced the number of read pairs from the majority sequence member that incorrectly aligned to the reference genome of the minority sequence member. Combining both source genomes into a single reference genome increased the specificity of mapping, while also reducing the central processing unit (CPU) time. In Plasmodium, at a seed length of 18 nt, 24.1 % of reads mapped to the human genome using 1.7±0.1 CPU hours, while 83.6 % of reads mapped to the Plasmodium genome using 0.2±0.0 CPU hours (total: 107.7 % reads mapping; in 1.9±0.1 CPU hours). In contrast, 97.1 % of the reads mapped to a combined Plasmodium-human reference in only 0.7±0.0 CPU hours. Overall, the results suggest that combining all references into a single reference database and using a 23 nt seed length reduces the computational time, while maximizing specificity. Similar results were found for simulated sequence reads from a mock metagenomic data set. We found similar improvements to computation time in a publicly available human-only data set.}, } @article {pmid29114059, year = {2017}, author = {Gomes, FM and Hixson, BL and Tyner, MDW and Ramirez, JL and Canepa, GE and Alves E Silva, TL and Molina-Cruz, A and Keita, M and Kane, F and Traoré, B and Sogoba, N and Barillas-Mury, C}, title = {Effect of naturally occurring Wolbachia in Anopheles gambiae s.l. mosquitoes from Mali on Plasmodium falciparum malaria transmission.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {114}, number = {47}, pages = {12566-12571}, pmid = {29114059}, issn = {1091-6490}, mesh = {Animals ; Anopheles/*microbiology/parasitology ; Female ; *Host-Parasite Interactions ; Host-Pathogen Interactions ; Insect Vectors/*microbiology/parasitology ; Malaria, Falciparum/epidemiology/parasitology/pathology/*transmission ; Mali/epidemiology ; Oocysts/pathogenicity/physiology ; Phylogeny ; Plasmodium falciparum/*microbiology ; RNA, Ribosomal, 16S/genetics ; Severity of Illness Index ; Sporozoites/pathogenicity/physiology ; Wolbachia/classification/*genetics/isolation & purification ; }, abstract = {A naturally occurring Wolbachia strain (wAnga-Mali) was identified in mosquitoes of the Anopheles gambiae complex collected in the Malian villages of Dangassa and Kenieroba. Phylogenetic analysis of the nucleotide sequence of two 16S rRNA regions showed that wAnga-Mali clusters with Wolbachia strains from supergroup A and has the highest homology to a Wolbachia strain isolated from cat fleas (Ctenocephalides). wAnga-Mali is different from two Wolbachia strains previously reported in A. gambiae from Burkina Faso (wAnga_VK5_STP and wAnga_VK5_3.1a). Quantitative analysis of Wolbachia and Plasmodium sporozoite infection in field-collected mosquitoes indicates that the prevalence and intensity of Plasmodium falciparum sporozoite infection is significantly lower in Wolbachia-infected females. The presence of Wolbachia in females from a laboratory Anopheles coluzzii (A. gambiae, M form) colony experimentally infected with P. falciparum (NF54 strain) gametocyte cultures slightly enhanced oocyst infection. However, Wolbachia infection significantly reduced the prevalence and intensity of sporozoite infection, as observed in the field. This indicates that wAnga-Mali infection does not limit early stages of Plasmodium infection in the mosquito, but it has a strong deleterious effect on sporozoites and reduces malaria transmission.}, } @article {pmid29106651, year = {2018}, author = {Sosa, EJ and Burguener, G and Lanzarotti, E and Defelipe, L and Radusky, L and Pardo, AM and Marti, M and Turjanski, AG and Fernández Do Porto, D}, title = {Target-Pathogen: a structural bioinformatic approach to prioritize drug targets in pathogens.}, journal = {Nucleic acids research}, volume = {46}, number = {D1}, pages = {D413-D418}, pmid = {29106651}, issn = {1362-4962}, mesh = {Amino Acid Sequence ; Anti-Infective Agents/*chemistry/pharmacology ; Binding Sites ; Communicable Diseases/drug therapy ; Computational Biology/*methods ; *Databases, Factual ; Drug Discovery ; *Genome, Bacterial ; *Genome, Fungal ; *Genome, Helminth ; *Genome, Protozoan ; Humans ; Internet ; Metabolic Networks and Pathways/drug effects/genetics ; Models, Molecular ; Molecular Targeted Therapy ; Protein Binding ; Protein Conformation, alpha-Helical ; Protein Conformation, beta-Strand ; Protein Interaction Domains and Motifs ; Sequence Alignment ; Sequence Homology, Amino Acid ; Software ; }, abstract = {Available genomic data for pathogens has created new opportunities for drug discovery and development to fight them, including new resistant and multiresistant strains. In particular structural data must be integrated with both, gene information and experimental results. In this sense, there is a lack of an online resource that allows genome wide-based data consolidation from diverse sources together with thorough bioinformatic analysis that allows easy filtering and scoring for fast target selection for drug discovery. Here, we present Target-Pathogen database (http://target.sbg.qb.fcen.uba.ar/patho), designed and developed as an online resource that allows the integration and weighting of protein information such as: function, metabolic role, off-targeting, structural properties including druggability, essentiality and omic experiments, to facilitate the identification and prioritization of candidate drug targets in pathogens. We include in the database 10 genomes of some of the most relevant microorganisms for human health (Mycobacterium tuberculosis, Mycobacterium leprae, Klebsiella pneumoniae, Plasmodium vivax, Toxoplasma gondii, Leishmania major, Wolbachia bancrofti, Trypanosoma brucei, Shigella dysenteriae and Schistosoma Smanosoni) and show its applicability. New genomes can be uploaded upon request.}, } @article {pmid29099918, year = {2017}, author = {Chung, M and Small, ST and Serre, D and Zimmerman, PA and Dunning Hotopp, JC}, title = {Draft genome sequence of the Wolbachia endosymbiont of Wuchereria bancrofti wWb.}, journal = {Pathogens and disease}, volume = {75}, number = {9}, pages = {}, pmid = {29099918}, issn = {2049-632X}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; T32 AI007024/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cluster Analysis ; Genes, rRNA ; *Genome, Bacterial ; Open Reading Frames ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 23S/genetics ; RNA, Ribosomal, 5S/genetics ; *Sequence Analysis, DNA ; Wolbachia/*genetics/isolation & purification ; Wuchereria bancrofti/*microbiology ; }, abstract = {The draft genome assembly of the Wolbachia endosymbiont of Wuchereria bancrofti (wWb) consists of 1060 850 bp in 100 contigs and contains 961 ORFs, with a single copy of the 5S rRNA, 16S rRNA and 23S rRNA and each of the 34 tRNA genes. Phylogenetic core genome analyses show wWb to cluster with other strains in supergroup D of the Wolbachia phylogeny, while being most closely related to the Wolbachia endosymbiont of Brugia malayi strain TRS (wBm). The wWb and wBm genomes share 779 orthologous clusters with wWb having 101 unclustered genes and wBm having 23 unclustered genes. The higher number of unclustered genes in the wWb genome likely reflects the fragmentation of the draft genome.}, } @article {pmid29099491, year = {2018}, author = {Pan, X and Pike, A and Joshi, D and Bian, G and McFadden, MJ and Lu, P and Liang, X and Zhang, F and Raikhel, AS and Xi, Z}, title = {The bacterium Wolbachia exploits host innate immunity to establish a symbiotic relationship with the dengue vector mosquito Aedes aegypti.}, journal = {The ISME journal}, volume = {12}, number = {1}, pages = {277-288}, pmid = {29099491}, issn = {1751-7370}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/immunology/*microbiology ; Animals ; *Immunity, Innate ; Mosquito Vectors/immunology/*microbiology ; Symbiosis/*physiology ; Toll-Like Receptors/metabolism ; Wolbachia/*physiology ; }, abstract = {A host's immune system plays a central role in shaping the composition of the microbiota and, in return, resident microbes influence immune responses. Symbiotic associations of the maternally transmitted bacterium Wolbachia occur with a wide range of arthropods. It is, however, absent from the dengue and Zika vector mosquito Aedes aegypti in nature. When Wolbachia is artificially forced to form symbiosis with this new mosquito host, it boosts the basal immune response and enhances the mosquito's resistance to pathogens, including dengue, Zika virus and malaria parasites. The mechanisms involved in establishing a symbiotic relationship between Wolbachia and A. aegypti, and the long-term outcomes of this interaction, are not well understood. Here, we have demonstrated that both the immune deficiency (IMD) and Toll pathways are activated by the Wolbachia strain wAlbB upon its introduction into A. aegypti. Silencing the Toll and IMD pathways via RNA interference reduces the wAlbB load. Notably, wAlbB induces peptidoglycan recognition protein (PGRP)-LE expression in the carcass of A. aegypti, and its silencing results in a reduction of symbiont load. Using transgenic mosquitoes with stage-specific induction of the IMD and Toll pathways, we have shown that elevated wAlbB infection in these mosquitoes is maintained via maternal transmission. These results indicate that host innate immunity is utilized to establish and promote host-microbial symbiosis. Our results will facilitate a long-term projection of the stability of the Wolbachia-A. aegypti mosquito system that is being developed to control dengue and Zika virus transmission to humans.}, } @article {pmid29095113, year = {2018}, author = {Moghadam, NN and Thorshauge, PM and Kristensen, TN and de Jonge, N and Bahrndorff, S and Kjeldal, H and Nielsen, JL}, title = {Strong responses of Drosophila melanogaster microbiota to developmental temperature.}, journal = {Fly}, volume = {12}, number = {1}, pages = {1-12}, pmid = {29095113}, issn = {1933-6942}, mesh = {Acetobacter/growth & development/isolation & purification ; Animals ; Drosophila melanogaster/*growth & development/*microbiology ; Female ; Gastrointestinal Microbiome ; Leuconostoc/growth & development/isolation & purification ; Male ; Temperature ; Wolbachia/growth & development/isolation & purification ; }, abstract = {Physiological responses to changes in environmental conditions such as temperature may partly arise from the resident microbial community that integrates a wide range of bio-physiological aspects of the host. In the present study, we assessed the effect of developmental temperature on the thermal tolerance and microbial community of Drosophila melanogaster. We also developed a bacterial transplantation protocol in order to examine the possibility of reshaping the host bacterial composition and assessed its influence on the thermotolerance phenotype. We found that the temperature during development affected thermal tolerance and the microbial composition of male D. melanogaster. Flies that developed at low temperature (13°C) were the most cold resistant and showed the highest abundance of Wolbachia, while flies that developed at high temperature (31°C) were the most heat tolerant and had the highest abundance of Acetobacter. In addition, feeding newly eclosed flies with bacterial suspensions from intestines of flies developed at low temperatures changed the heat tolerance of recipient flies. However, we were not able to link this directly to a change in the host bacterial composition.}, } @article {pmid29094498, year = {2019}, author = {Pierzynowska, K and Skowron Volponi, M and Węgrzyn, G}, title = {Multiple factors correlating with wing malformations in the population of Parnassius apollo (Lepidoptera: Papilionidae) restituted from a low number of individuals: A mini review.}, journal = {Insect science}, volume = {26}, number = {3}, pages = {380-387}, doi = {10.1111/1744-7917.12554}, pmid = {29094498}, issn = {1744-7917}, support = {530-L140-D242-17-1A//University of Gdansk/ ; }, mesh = {Animals ; *Butterflies ; Conservation of Natural Resources ; *Endangered Species ; Insect Proteins/genetics ; Laccase/metabolism ; Wings, Animal/*abnormalities ; Wolbachia ; Yersinia ; }, abstract = {The Apollo butterfly, Parnassius apollo (Linnaeus), was common in Europe over 100 years ago, but currently it is considered as near threatened. Different conservation programs have promoted the persistence of this species; however, it is still endangered. An example of such programs was the action devoted to reestablish the Apollo butterfly population in Pieniny National Park (Poland) from only 20-30 individuals which had survived till the last decade of the 20th century. This reintroduction has been successful; however, unexpected developmental problems appeared. Butterflies with deformed or reduced wings became frequent in the population living in the natural habitat, and particularly among those reared under seminatural conditions (in the same environment, but fenced by a net). Until recently, reasons for these malformations remained unknown. However, reports published during last months indicated that there are genetic, biochemical, and microbiological factors contributing to this phenomenon. In the malformed individuals, lesions in the wingless gene and dysfunctions of laccase 1 and 2 were found to be significantly more frequent than in normal insects. A large fraction of butterflies with deformed or reduced wings was devoid of the prokaryotic symbiont Wolbachia, which was present in most normal individuals. Moreover, Yersinia pseudotuberculosis (Pfeiffer) Smith and Thal, and Serratia sp., bacteria pathogenic to insects, were detected in the biological material from both normal and malformed butterflies from this population. These findings are summarized and discussed in this review, in the light of conservation of insects and restitution of their populations from a low number of individuals.}, } @article {pmid29093481, year = {2017}, author = {Karakuş, M and Karabey, B and Orçun Kalkan, Ş and Özdemir, G and Oğuz, G and Erişöz Kasap, Ö and Alten, B and Töz, S and Özbel, Y}, title = {Midgut Bacterial Diversity of Wild Populations of Phlebotomus (P.) papatasi, the Vector of Zoonotic Cutaneous Leishmaniasis (ZCL) in Turkey.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {14812}, pmid = {29093481}, issn = {2045-2322}, mesh = {Animals ; Bacteria/genetics/*isolation & purification ; Female ; Gastrointestinal Microbiome ; Humans ; Insect Vectors/*microbiology ; Leishmaniasis, Cutaneous/*epidemiology ; Male ; Phlebotomus/*microbiology ; Proteobacteria/genetics/isolation & purification ; Turkey/epidemiology ; Wolbachia/genetics/isolation & purification ; }, abstract = {Phlebotomine sand flies are hematophagous insects that harbor bacterial, viral and parasitic agents like Bartonella sp., Phleboviruses and Leishmania spp., respectively. There are few reports on bacterial microbiota of Phlebotomus (P.) papatasi but no data available for natural populations of Turkey, where leishmaniasis is endemic. Therefore, we aimed to investigate the midgut bacterial flora of different populations of P. papatasi. Sand flies were collected from different towns (Karaburun, Urla, Ayvacik and Başçayır) located in the western part of Turkey. Laboratory reared P. papatasi were included in the study as an insectarium population. After sterile washing steps, sand flies were dissected and guts were separated. Three pools, (males, unfed females and blood-fed females) were generated for each population. Prokaryotic 16 S rRNA gene was amplified and DGGE was performed. Fourteen different organisms belonging to two Phylum (Proteobactericea and Furmicutes) were identified according to sequence results in the studied pools. The presence of Wolbachia sp. was shown for the first time in the wild-caught sand fly populations of Turkey. This is the first report of gut bacterial flora of wild-caught P. papatasi collected in an endemic area for leishmaniasis in Turkey. Microbiome profiling of wild-caught sand flies will be of great help in the investigating of possible vector control candidates for paratransgenic control approach.}, } @article {pmid29093474, year = {2017}, author = {Kaur, R and Siozios, S and Miller, WJ and Rota-Stabelli, O}, title = {Insertion sequence polymorphism and genomic rearrangements uncover hidden Wolbachia diversity in Drosophila suzukii and D. subpulchrella.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {14815}, pmid = {29093474}, issn = {2045-2322}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; DNA Transposable Elements ; Drosophila/*microbiology/physiology ; Fertility ; Multilocus Sequence Typing ; Phylogeny ; *Polymorphism, Genetic ; Wolbachia/classification/*genetics ; }, abstract = {Ability to distinguish between closely related Wolbachia strains is crucial for understanding the evolution of Wolbachia-host interactions and the diversity of Wolbachia-induced phenotypes. A useful model to tackle these issues is the Drosophila suzukii - Wolbachia association. D. suzukii, a destructive insect pest, harbor a non-CI inducing Wolbachia 'wSuz' closely related to the strong CI-inducing wRi strain. Multi locus sequence typing (MLST) suggests presence of genetic homogeneity across wSuz strains infecting European and American D. suzukii populations, although different Wolbachia infection frequencies and host fecundity levels have been observed in both populations. Currently, it is not clear if these differences are due to cryptic wSuz polymorphism, host background, geographical factors or a combination of all of them. Here, we have identified geographical diversity in wSuz in D. suzukii populations from different continents using a highly diagnostic set of markers based on insertion sequence (IS) site polymorphism and genomic rearrangements (GR). We further identified inter-strain diversity between Wolbachia infecting D. suzukii and its sister species D. subpulchrella (wSpc). Based on our results, we speculate that discernible wSuz variants may associate with different observed host phenotypes, a hypothesis that demands future investigation. More generally, our results demonstrate the utility of IS and GRs in discriminating closely related Wolbachia strains.}, } @article {pmid29084217, year = {2017}, author = {Dodson, BL and Andrews, ES and Turell, MJ and Rasgon, JL}, title = {Wolbachia effects on Rift Valley fever virus infection in Culex tarsalis mosquitoes.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {10}, pages = {e0006050}, pmid = {29084217}, issn = {1935-2735}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; R21 AI128918/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culex/*microbiology/*virology ; Host-Pathogen Interactions ; Insect Vectors/microbiology/virology ; Mosquito Control ; Pest Control, Biological/*methods ; Rift Valley fever virus/*physiology ; Wolbachia/*physiology ; }, abstract = {Innovative tools are needed to alleviate the burden of mosquito-borne diseases, and strategies that target the pathogen are being considered. A possible tactic is the use of Wolbachia, a maternally inherited, endosymbiotic bacterium that can (but does not always) suppress diverse pathogens when introduced to naive mosquito species. We investigated effects of somatic Wolbachia (strain wAlbB) infection on Rift Valley fever virus (RVFV) in Culex tarsalis mosquitoes. When compared to Wolbachia-uninfected mosquitoes, there was no significant effect of Wolbachia infection on RVFV infection, dissemination, or transmission frequencies, nor on viral body or saliva titers. Within Wolbachia-infected mosquitoes, there was a modest negative correlation between RVFV body titers and Wolbachia density, suggesting that Wolbachia may slightly suppress RVFV in a density-dependent manner in this mosquito species. These results are contrary to previous work in the same mosquito species, showing Wolbachia-induced enhancement of West Nile virus infection rates. Taken together, these results highlight the importance of exploring the breadth of pathogen modulations induced by Wolbachia.}, } @article {pmid29080766, year = {2017}, author = {Diosdado, A and Gómez, PJ and Morchón, R and Simón, F and González-Miguel, J}, title = {Interaction between Wolbachia and the fibrinolytic system as a possible pathological mechanism in cardiopulmonary dirofilariosis.}, journal = {Veterinary parasitology}, volume = {247}, number = {}, pages = {64-69}, doi = {10.1016/j.vetpar.2017.10.001}, pmid = {29080766}, issn = {1873-2550}, mesh = {Amino Acid Sequence ; Animals ; Dirofilaria immitis/*microbiology ; Dirofilariasis/*parasitology ; Dog Diseases/*parasitology ; Dogs ; Fibrinolysin/metabolism ; Fibrinolysis ; Host-Parasite Interactions ; Humans ; Membrane Proteins/genetics/*metabolism ; Models, Molecular ; Recombinant Proteins ; Sequence Alignment/veterinary ; Symbiosis ; Tissue Plasminogen Activator/*metabolism ; Wolbachia/*physiology ; }, abstract = {Dirofilaria immitis is a mosquito-borne parasite that produces an inflammatory process in the wall of the blood vessels of its definitive host during cardiopulmonary dirofilariosis, known as proliferative endarteritis. Parasite antigens participate in the appearance of this inflammatory event, among other mechanisms through the over-activation of the host fibrinolytic system. Since Wolbachia, endosymbiont bacteria of filarial nematodes, is released into the vertebrate host when worms die, the aim of this work was to analyse the interaction between this bacteria and the host fibrinolytic system to complete the study of this part of the host-parasite relationships. For that purpose, the recombinant form of the major Wolbachia surface protein (rWSP) was cloned, sequenced and expressed and then, its ability to bind plasminogen and enhance the generation of plasmin was assessed. We demonstrated that rWSP is a conserved antigen within the family Onchocercidae with ability to bind plasminogen and stimulate plasmin generation in a tissue-plasminogen activator (t-PA) and lysine residues of the rWSP-dependent manner. These results indicate that the recruitment of plasminogen by Wolbachia and the possible excess of plasmin generated could contribute to exacerbate the pathological events occurred at the vascular level during cardiopulmonary dirofilariosis, as well as in other diseases caused by filarial nematodes that harbour Wolbachia, when the bacteria is released after the death of the worms.}, } @article {pmid29078351, year = {2017}, author = {Turner, JD and Sharma, R and Al Jayoussi, G and Tyrer, HE and Gamble, J and Hayward, L and Priestley, RS and Murphy, EA and Davies, J and Waterhouse, D and Cook, DAN and Clare, RH and Cassidy, A and Steven, A and Johnston, KL and McCall, J and Ford, L and Hemingway, J and Ward, SA and Taylor, MJ}, title = {Albendazole and antibiotics synergize to deliver short-course anti-Wolbachia curative treatments in preclinical models of filariasis.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {114}, number = {45}, pages = {E9712-E9721}, pmid = {29078351}, issn = {1091-6490}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Albendazole/*pharmacology ; Animals ; Anti-Bacterial Agents/*pharmacology ; Benzimidazoles/pharmacology ; Brugia malayi/microbiology ; Drug Synergism ; Female ; Filariasis/*drug therapy ; Male ; Mice ; Mice, Inbred BALB C ; Minocycline/pharmacology ; Rifampin/pharmacology ; Wolbachia/*drug effects ; }, abstract = {Elimination of filariasis requires a macrofilaricide treatment that can be delivered within a 7-day period. Here we have identified a synergy between the anthelmintic albendazole (ABZ) and drugs depleting the filarial endosymbiont Wolbachia, a proven macrofilaricide target, which reduces treatment from several weeks to 7 days in preclinical models. ABZ had negligible effects on Wolbachia but synergized with minocycline or rifampicin (RIF) to deplete symbionts, block embryogenesis, and stop microfilariae production. Greater than 99% Wolbachia depletion following 7-day combination of RIF+ABZ also led to accelerated macrofilaricidal activity. Thus, we provide preclinical proof-of-concept of treatment shortening using antibiotic+ABZ combinations to deliver anti-Wolbachia sterilizing and macrofilaricidal effects. Our data are of immediate public health importance as RIF+ABZ are registered drugs and thus immediately implementable to deliver a 1-wk macrofilaricide. They also suggest that novel, more potent anti-Wolbachia drugs under development may be capable of delivering further treatment shortening, to days rather than weeks, if combined with benzimidazoles.}, } @article {pmid29076805, year = {2018}, author = {Lourenço-de-Oliveira, R and Marques, JT and Sreenu, VB and Atyame Nten, C and Aguiar, ERGR and Varjak, M and Kohl, A and Failloux, AB}, title = {Culex quinquefasciatus mosquitoes do not support replication of Zika virus.}, journal = {The Journal of general virology}, volume = {99}, number = {2}, pages = {258-264}, pmid = {29076805}, issn = {1465-2099}, support = {G0801822/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/12/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; U01 AI115595/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culex/*virology ; Female ; Mosquito Vectors/*virology ; Phenotype ; *Virus Replication ; Wolbachia/*physiology ; Zika Virus/*physiology ; Zika Virus Infection/*transmission/virology ; }, abstract = {The rapid spread of Zika virus (ZIKV) in the Americas raised many questions about the role of Culex quinquefasciatus mosquitoes in transmission, in addition to the key role played by the vector Aedes aegypti. Here we analysed the competence of Cx. quinquefasciatus (with or without Wolbachia endosymbionts) for a ZIKV isolate. We also examined the induction of RNA interference pathways after viral challenge and the production of small virus-derived RNAs. We did not observe any infection nor such small virus-derived RNAs, regardless of the presence or absence of Wolbachia. Thus, Cx. quinquefasciatus does not support ZIKV replication and Wolbachia is not involved in producing this phenotype. In short, these mosquitoes are very unlikely to play a role in transmission of ZIKV.}, } @article {pmid29076641, year = {2017}, author = {Bhattacharya, T and Newton, ILG}, title = {Mi Casa es Su Casa: how an intracellular symbiont manipulates host biology.}, journal = {Environmental microbiology}, volume = {}, number = {}, pages = {}, pmid = {29076641}, issn = {1462-2920}, support = {R21 AI121849/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia pipientis, the most common intracellular infection on the planet, infects 40% of insects as well as nematodes, isopods and arachnids. Wolbachia are obligately intracellular and challenging to study; there are no genetic tools for manipulating Wolbachia nor can they be cultured outside of host cells. Despite these roadblocks, the research community has defined a set of Wolbachia loci involved in host interaction: Wolbachia effectors. Through the use of Drosophila genetics, surrogate systems and biochemistry, the field has begun to define the toolkit Wolbachia use for host manipulation. Below we review recent findings identifying these Wolbachia effectors and point to potential, as yet uncharacterized, links between known phenotypes induced by Wolbachia infection and predicted effectors.}, } @article {pmid29074653, year = {2017}, author = {Ward, CM and Baxter, SW}, title = {Draft Genome Assembly of a Wolbachia Endosymbiont of Plutella australiana.}, journal = {Genome announcements}, volume = {5}, number = {43}, pages = {}, pmid = {29074653}, issn = {2169-8287}, abstract = {Wolbachia spp. are endosymbiotic bacteria that infect around 50% of arthropods and cause a broad range of effects, including manipulating host reproduction. Here, we present the annotated draft genome assembly of Wolbachia strain wAus, which infects Plutella australiana, a cryptic ally of the major Brassica pest Plutella xylostella (diamondback moth).}, } @article {pmid29071168, year = {2017}, author = {Zhang, L and Zhang, G and Yun, Y and Peng, Y}, title = {Bacterial community of a spider, Marpiss magister (Salticidae).}, journal = {3 Biotech}, volume = {7}, number = {6}, pages = {371}, pmid = {29071168}, issn = {2190-572X}, abstract = {Arthropods are associated with various microorganisms which confer benefits to their hosts. Recently, research has been conducted on bacterial communities of insects to provide an insight into the potential interactions of the symbiotic bacteria and their hosts. Spiders are interesting to study as they are perceived to be natural enemies of pests. The effect of endosymbionts on spiders has been reported, but little is known about the overall bacterial communities present in spiders. Here, we report on the characterization of bacterial communities present in the whole body of the spider Marpiss magister using Illumina sequencing of 16S rRNA amplicons. Our study shows that the most abundant phyla of bacteria included Proteobacteria, Tenericutes, Bacteroidetes and Actinobacteria. At the genus level, the most abundant genera included Rickettsia, Wolbachia, Spiroplasma, and Cardinium. Besides these dominant endosymbionts, our study also showed the existence of bacteria in the genera Arthrobacter, Novosphingobium, Acinetobacter, Pseudomonas, Aquabacterium and Sphingomonas at an abundance ranging from 0.65 to 0.84%, and the existence of bacterial in genera Lactobacillus, Sphingobium, Methylobacterium, Bradyrhizobium, Propionibacterium, Brevundimonas, Achromobacter, Microbacterium, Corynebacterium, and Flavobacterium at a slightly lower abundance ranging from 0.1 to 0.5%. Therefore, our finding indicates that endosymbionts are not the only microbiota present in the spider M. magister, and other bacterial taxa also exist in its bacterial community.}, } @article {pmid29067008, year = {2017}, author = {Zhukova, M and Sapountzis, P and Schiøtt, M and Boomsma, JJ}, title = {Diversity and Transmission of Gut Bacteria in Atta and Acromyrmex Leaf-Cutting Ants during Development.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {1942}, pmid = {29067008}, issn = {1664-302X}, support = {323085/ERC_/European Research Council/International ; }, abstract = {The social Hymenoptera have distinct larval and adult stages separated by metamorphosis, which implies striking remodeling of external and internal body structures during the pupal stage. This imposes challenges to gut symbionts as existing cultures are lost and may or may not need to be replaced. To elucidate the extent to which metamorphosis interrupts associations between bacteria and hosts, we analyzed changes in gut microbiota during development and traced the transmission routes of dominant symbionts from the egg to adult stage in the leaf-cutting ants Acromyrmex echinatior and Atta cephalotes, which are both important functional herbivores in the New World tropics. Bacterial density remained similar across the developmental stages of Acromyrmex, but Atta brood had very low bacterial prevalences suggesting that bacterial gut symbionts are not actively maintained. We found that Wolbachia was the absolute dominant bacterial species across developmental stages in Acromyrmex and we confirmed that Atta lacks Wolbachia also in the immature stages, and had mostly Mollicutes bacteria in the adult worker guts. Wolbachia in Acromyrmex appeared to be transovarially transmitted similar to transmission in solitary insects. In contrast, Mollicutes were socially transmitted from old workers to newly emerged callows. We found that larval and pupal guts of both ant species contained Pseudomonas and Enterobacter bacteria that are also found in fungus gardens, but hardly or not in adult workers, suggesting they are beneficial only for larval growth and development. Our results reveal that transmission pathways for bacterial symbionts may be very different both between developmental stages and between sister genera and that identifying the mechanisms of bacterial acquisition and loss will be important to clarify their putative mutualistic functions.}, } @article {pmid29066721, year = {2017}, author = {He, S and Zhang, X and Liang, J and Tang, S}, title = {Multiscale modelling the effects of CI genetic evolution in mosquito population on the control of dengue fever.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {13895}, pmid = {29066721}, issn = {2045-2322}, mesh = {Animals ; Culicidae/*genetics/microbiology/*physiology ; Cytoplasm/*genetics ; Dengue/*prevention & control/transmission ; Evolution, Molecular ; Female ; Gene Frequency ; Genotype ; Male ; *Models, Genetic ; Mosquito Control/*methods ; Reproduction/*genetics ; Wolbachia/genetics ; }, abstract = {Endosymbiotic Wolbachia bacteria are widely applied for the control of dengue fever by manipulating the reproductive mechanism of mosquitoes, including maternal inheritance and cytoplasmic incompatibility (CI). CI means that the offsprings from the matings between Wolbachia infected males and uninfected females can not be hatched. At present, CI effect is assumed as a constant in most of dynamic systems for the spread of Wolbachia. However, their spread may arouse the evolution of mosquitoes to resist CI. Thus, a multiscale model combining a birth-pulse model with a gene-induced discrete model for the frequencies of alleles is proposed to describe the spread of Wolbachia in mosquito population with resistance allele of CI. The main results indicate that the strategy of population eradication can not be realized, while the strategy of population replacement may be realized with the success of sensitive or resistance allele. If appropriate Wolbachia strains can not be selected, then there is a high probability of the failure of population replacement. Moreover, Wolbachia-induced parameters may arouse the catastrophic shifts among stable states of the model. In addition, the demographic parameters and Wolbachia-induced parameters may affect the level and the speed of population replacement and the density of uninfected mosquitoes.}, } @article {pmid29065795, year = {2017}, author = {Al-Abd, NM and Nor, ZM and Junaid, QO and Mansor, M and Hasan, MS and Kassim, M}, title = {Antifilarial activity of caffeic acid phenethyl ester on Brugia pahangi in vitro and in vivo.}, journal = {Pathogens and global health}, volume = {111}, number = {7}, pages = {388-394}, pmid = {29065795}, issn = {2047-7732}, mesh = {Animals ; Brugia pahangi/*drug effects ; Caffeic Acids/administration & dosage/*therapeutic use ; Dose-Response Relationship, Drug ; Filariasis/*drug therapy/parasitology ; Gerbillinae ; Humans ; Male ; Phenylethyl Alcohol/administration & dosage/*analogs & derivatives/therapeutic use ; Wolbachia/drug effects ; }, abstract = {Lymphatic filariasis (LF) is a vector borne disease caused by parasitic worms such as Wuchereria bancrofti, Brugia malayi and B. timori, which are transmitted by mosquitoes. Current therapeutics to treat LF are mainly microfilarcidal, and lack activity against adult worms. This set back, poses a challenge for the control and elimination of filariasis. Thus, in this study the activities of caffeic acid phenethyl ester (CAPE) against the filarial worm B. pahangi and its bacterial endosymbiont, Wolbachia were evaluated. Different concentrations (2, 5, 10, 15, 20 μg/ml) of CAPE were used to assess its effects on motility, viability and microfilarial (mf) production of B. pahangi in vitro. Anti-Wolbachial activity of CAPE was measured in worms by quantification of Wolbachial wsp gene copy number using real-time polymerase chain reaction. Our findings show that CAPE was found to significantly reduce adult worm motility, viability, and mf release both in vitro and in vivo. 20 μg/ml of CAPE halts the release of mf in vitro by day 6 of post treatment. Also, the number of adult worms recovered in vivo were reduced significantly during and after treatment with 50 mg/kg of CAPE relative to control drugs, diethylcarbamazine and doxycycline. Real time PCR based on the Wolbachia ftsZ gene revealed a significant reduction in Wolbachia copy number upon treatment. Anti-Wolbachia and antifilarial properties of CAPE require further investigation as an alternative strategy to treat LF.}, } @article {pmid29063344, year = {2018}, author = {Hornok, S and Ágh, N and Takács, N and Kontschán, J and Hofmann-Lehmann, R}, title = {Haematospirillum and insect Wolbachia DNA in avian blood.}, journal = {Antonie van Leeuwenhoek}, volume = {111}, number = {3}, pages = {479-483}, doi = {10.1007/s10482-017-0961-0}, pmid = {29063344}, issn = {1572-9699}, support = {115854//OTKA/ ; 108571//OTKA/ ; }, mesh = {Alphaproteobacteria/*genetics ; Animals ; Birds/*blood ; *DNA, Bacterial ; Female ; *Genes, Insect ; Male ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {In this study, blood samples of 259 Acrocephalus sp. warblers were molecularly analysed for Anaplasmataceae and Rhodospirillaceae based on PCR amplification of 16S rRNA gene fragments. One bird blood sample (from Reed Warbler, Acrocephalus scirpaceus) yielded a sequence with 99.8% identity to Haematospirillum jordaniae. This is the first molecular evidence for the occurrence of this species in the blood of any vertebrate other than human. Another bird blood sample (from Marsh Warbler: Acrocephalus palustris) yielded a Wolbachia sequence, closely related to a moth endosymbiont with 99.8% identity. A nematode origin of Wolbachia DNA detected here in avian blood can be excluded, because results of phylogenetic analysis showed its closest alignment with insect wolbachiae. This is the first finding of insect Wolbachia DNA in the circulatory system of birds, which can be explained either by the inoculation of wolbachiae by blood-sucking vectors, or passing of Wolbachia DNA from the gut into the blood of this insectivorous bird species.}, } @article {pmid29053633, year = {2017}, author = {Skaljac, M and Kanakala, S and Zanic, K and Puizina, J and Pleic, IL and Ghanim, M}, title = {Diversity and Phylogenetic Analyses of Bacterial Symbionts in Three Whitefly Species from Southeast Europe.}, journal = {Insects}, volume = {8}, number = {4}, pages = {}, pmid = {29053633}, issn = {2075-4450}, abstract = {Bemisia tabaci (Gennadius), Trialeurodes vaporariorum (Westwood), and Siphoninus phillyreae (Haliday) are whitefly species that harm agricultural crops in many regions of the world. These insects live in close association with bacterial symbionts that affect host fitness and adaptation to the environment. In the current study, we surveyed the infection of whitefly populations in Southeast Europe by various bacterial symbionts and performed phylogenetic analyses on the different symbionts detected. Arsenophonus and Hamiltonella were the most prevalent symbionts in all three whitefly species. Rickettsia was found to infect mainly B. tabaci, while Wolbachia mainly infected both B. tabaci and S. phillyreae. Furthermore, Cardinium was rarely found in the investigated whitefly populations, while Fritschea was never found in any of the whitefly species tested. Phylogenetic analyses revealed a diversity of several symbionts (e.g., Hamiltonella, Arsenophonus, Rickettsia), which appeared in several clades. Reproductively isolated B. tabaci and T. vaporariorum shared the same (or highly similar) Hamiltonella and Arsenophonus, while these symbionts were distinctive in S. phillyreae. Interestingly, Arsenophonus from S. phillyreae did not cluster with any of the reported sequences, which could indicate the presence of Arsenophonus, not previously associated with whiteflies. In this study, symbionts (Wolbachia, Rickettsia, and Cardinium) known to infect a wide range of insects each clustered in the same clades independently of the whitefly species. These results indicate horizontal transmission of bacterial symbionts between reproductively isolated whitefly species, a mechanism that can establish new infections that did not previously exist in whiteflies.}, } @article {pmid29042606, year = {2017}, author = {Signor, S}, title = {Population genomics of Wolbachia and mtDNA in Drosophila simulans from California.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {13369}, pmid = {29042606}, issn = {2045-2322}, mesh = {California ; DNA, Mitochondrial/*genetics ; Drosophila simulans/*microbiology ; Evolution, Molecular ; Gene Dosage ; *Genetics, Population ; *Genomics/methods ; Linkage Disequilibrium ; Phylogeny ; Polymorphism, Single Nucleotide ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia pipientis is an intracellular endosymbiont infecting many arthropods and filarial nematodes. Little is known about the short-term evolution of Wolbachia or its interaction with its host. Wolbachia is maternally inherited, resulting in co-inheritance of mitochondrial organelles such as mtDNA. Here I explore the evolution of Wolbachia, and the relationship between Wolbachia and mtDNA, using a large inbred panel of Drosophila simulans. I compare this to the only other large population genomic Wolbachia dataset from D. melanogaster. I find reduced diversity relative to expectation in both Wolbachia and mtDNA, but only mtDNA shows evidence of a recent selective sweep or population bottleneck. I estimate Wolbachia and mtDNA titre in each genotype, and I find considerable variation in both phenotypes, despite low genetic diversity in Wolbachia and mtDNA. A phylogeny of Wolbachia and of mtDNA suggest a recent origin of the infection derived from a single origin. Using Wolbachia and mtDNA titre as a phenotype, I perform the first association analysis using this phenotype with the nuclear genome and find several implicated regions, including one which contains four CAAX-box protein processing genes. CAAX-box protein processing can be an important part of host-pathogen interactions in other systems, suggesting interesting directions for future research.}, } @article {pmid29036401, year = {2017}, author = {Santos-Garcia, D and Silva, FJ and Morin, S and Dettner, K and Kuechler, SM}, title = {The All-Rounder Sodalis: A New Bacteriome-Associated Endosymbiont of the Lygaeoid Bug Henestaris halophilus (Heteroptera: Henestarinae) and a Critical Examination of Its Evolution.}, journal = {Genome biology and evolution}, volume = {9}, number = {10}, pages = {2893-2910}, pmid = {29036401}, issn = {1759-6653}, mesh = {Animals ; DNA, Bacterial ; Databases, Factual ; Enterobacteriaceae/*classification/genetics/*physiology/ultrastructure ; *Evolution, Molecular ; Genome Size ; Genome, Bacterial ; Heteroptera/*microbiology ; Metabolic Networks and Pathways ; *Phylogeny ; Pseudogenes ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Hemipteran insects are well-known in their ability to establish symbiotic relationships with bacteria. Among them, heteropteran insects present an array of symbiotic systems, ranging from the most common gut crypt symbiosis to the more restricted bacteriome-associated endosymbiosis, which have only been detected in members of the superfamily Lygaeoidea and the family Cimicidae so far. Genomic data of heteropteran endosymbionts are scarce and have merely been analyzed from the Wolbachia endosymbiont in bed bug and a few gut crypt-associated symbionts in pentatomoid bugs. In this study, we present the first detailed genomic analysis of a bacteriome-associated endosymbiont of a phytophagous heteropteran, present in the seed bug Henestaris halophilus (Hemiptera: Heteroptera: Lygaeoidea). Using phylogenomics and genomics approaches, we have assigned the newly characterized endosymbiont to the Sodalis genus, named as Candidatus Sodalis baculum sp. nov. strain kilmister. In addition, our findings support the reunification of the Sodalis genus, currently divided into six different genera. We have also conducted comparative analyses between 15 Sodalis species that present different genome sizes and symbiotic relationships. These analyses suggest that Ca. Sodalis baculum is a mutualistic endosymbiont capable of supplying the amino acids tyrosine, lysine, and some cofactors to its host. It has a small genome with pseudogenes but no mobile elements, which indicates middle-stage reductive evolution. Most of the genes in Ca. Sodalis baculum are likely to be evolving under purifying selection with several signals pointing to the retention of the lysine/tyrosine biosynthetic pathways compared with other Sodalis.}, } @article {pmid29021316, year = {2017}, author = {Kageyama, D and Yoshimura, K and Sugimoto, TN and Katoh, TK and Watada, M}, title = {Maternally transmitted non-bacterial male killer in Drosophila biauraria.}, journal = {Biology letters}, volume = {13}, number = {10}, pages = {}, pmid = {29021316}, issn = {1744-957X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Drosophila/*physiology ; Female ; Japan ; Male ; Maternal Inheritance ; Phenotype ; Polymerase Chain Reaction ; Sex Ratio ; Symbiosis ; Tetracycline/pharmacology ; }, abstract = {A maternally inherited, all-female trait is widely found among arthropods, which is caused by bacterial endosymbionts such as Wolbachia, Rickettsia, Spiroplasma and Cardinium We discovered a single female of Drosophila biauraria, collected from Tomakomai, Hokkaido, Japan, that produced all-female offspring. This all-female trait was maternally inherited in the iso-female line (SP12F) by backcrossing with males of a normal line (SP11-20) with a 1 : 1 sex ratio derived from the same population. The all-female trait was not affected by tetracycline treatment performed for two consecutive generations. However, the microinjection of filter-sterilized homogenate of SP12F females into SP11-20 females established all-female matrilines. Our data suggest the role of transmissible agents, most likely viruses, but not bacteria or protists, as the possible cause of the all-female phenotype, which is likely to be achieved by killing of male embryos because egg hatch rates of SP12F were nearly half those of SP11-20. This is the first report in Diptera to demonstrate a maternally inherited virus-like element as the cause of the male-killing phenotype in D. biauraria.}, } @article {pmid28991902, year = {2017}, author = {de Oliveira, S and Villela, DAM and Dias, FBS and Moreira, LA and Maciel de Freitas, R}, title = {How does competition among wild type mosquitoes influence the performance of Aedes aegypti and dissemination of Wolbachia pipientis?.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {10}, pages = {e0005947}, pmid = {28991902}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Host-Pathogen Interactions ; Larva ; Pest Control, Biological/methods ; Population Density ; Species Specificity ; Survival Analysis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia has been deployed in several countries to reduce transmission of dengue, Zika and chikungunya viruses. During releases, Wolbachia-infected females are likely to lay their eggs in local available breeding sites, which might already be colonized by local Aedes sp. mosquitoes. Therefore, there is an urgent need to estimate the deleterious effects of intra and interspecific larval competition on mosquito life history traits, especially on the duration of larval development time, larval mortality and adult size.

Three different mosquito populations were used: Ae. aegypti infected with Wolbachia (wMelBr strain), wild Ae. aegypti and wild Ae. albopictus. A total of 21 treatments explored intra and interspecific larval competition with varying larval densities, species proportions and food levels. Each treatment had eight replicates with two distinct food levels: 0.25 or 0.50 g of Chitosan and fallen avocado leaves. Overall, overcrowding reduced fitness correlates of the three populations. Ae. albopictus larvae presented lower larval mortality, shorter development time to adult and smaller wing sizes than Ae. aegypti. The presence of Wolbachia had a slight positive effect on larval biology, since infected individuals had higher survivorship than uninfected Ae. aegypti larvae.

CONCLUSIONS/SIGNIFICANCE: In all treatments, Ae. albopictus outperformed both wild Ae. aegypti and the Wolbachia-infected group in larval competition, irrespective of larval density and the amount of food resources. The major force that can slow down Wolbachia invasion is the population density of wild mosquitoes. Given that Ae. aegypti currently dominates in Rio, in comparison with Ae. albopictus frequency, additional attention must be given to the population density of Ae. aegypti during releases to increase the likelihood of Wolbachia invasion.}, } @article {pmid28980839, year = {2018}, author = {Akmal, M and Freed, S and Dietrich, CH and Mehmood, M and Razaq, M}, title = {Patterns of genetic differentiation among populations of Amrasca biguttula biguttula (Shiraki) (Cicadellidae: Hemiptera).}, journal = {Mitochondrial DNA. Part A, DNA mapping, sequencing, and analysis}, volume = {29}, number = {6}, pages = {897-904}, doi = {10.1080/24701394.2017.1383405}, pmid = {28980839}, issn = {2470-1408}, mesh = {Animals ; Electron Transport Complex IV/*genetics ; Evolution, Molecular ; *Genetic Speciation ; Genome, Mitochondrial ; Hemiptera/classification/*genetics/microbiology ; India ; Insect Proteins/*genetics ; *Polymorphism, Genetic ; Wolbachia/pathogenicity ; }, abstract = {Cotton leafhopper, Amrasca biguttula biguttula (Shiraki), a serious sucking insect pest of cotton and vegetables is present throughout South and Southeast Asia. Genetic differentiation within A. biguttula biguttula populations collected from 16 cotton growing areas of Punjab, Pakistan, was examined by sequencing the barcode region of the mitochondrial cytochrome oxidase subunit 1 (COI) gene. The dendrogram obtained by neighbour joining analysis of COI sequences confirmed the presence of single species of cotton leafhopper. The overall average pairwise divergence was 0.01. Very little variation was found among populations from cotton growing areas of Punjab, Pakistan and these were most similar to populations from North India. South Indian populations were grouped together and were generally more divergent. Extensive migration of this pest species among cotton-growing areas in the Indian subcontinent may hinder genetic diversification of cotton leafhopper. Four Pakistani samples of cotton leafhopper tested positive for Wolbachia infection but were not clearly differentiated from non-Wolbachia infected samples, suggesting that Wolbachia did not cause reproductive incompatibilities.}, } @article {pmid28977708, year = {2017}, author = {Cariou, M and Duret, L and Charlat, S}, title = {The global impact of Wolbachia on mitochondrial diversity and evolution.}, journal = {Journal of evolutionary biology}, volume = {30}, number = {12}, pages = {2204-2210}, doi = {10.1111/jeb.13186}, pmid = {28977708}, issn = {1420-9101}, mesh = {Animals ; Arthropods/*genetics/microbiology ; *Biological Evolution ; DNA, Mitochondrial/*genetics ; Female ; Genetic Variation ; Male ; Phylogeny ; Wolbachia/*physiology ; }, abstract = {The spread of maternally inherited microorganisms, such as Wolbachia bacteria, can induce indirect selective sweeps on host mitochondria, to which they are linked within the cytoplasm. The resulting reduction in effective population size might lead to smaller mitochondrial diversity and reduced efficiency of natural selection. While documented in several host species, it is currently unclear if such a scenario is common enough to globally impact the diversity and evolution of mitochondria in Wolbachia-infected lineages. Here, we address this question using a mapping of Wolbachia acquisition/extinction events on a large mitochondrial DNA tree, including over 1000 species. Our analyses indicate that on a large phylogenetic scale, other sources of variation, such as mutation rates, tend to hide the effects of Wolbachia. However, paired comparisons between closely related infected and uninfected taxa reveal that Wolbachia is associated with a twofold reduction in silent mitochondrial polymorphism, and a 13% increase in nonsynonymous substitution rates. These findings validate the conjecture that the widespread distribution of Wolbachia infections throughout arthropods impacts the effective population size of mitochondria. These effects might in part explain the disconnection between genetic diversity and demographic population size in mitochondria, and also fuel red-queen-like cytonuclear co-evolution through the fixation of deleterious mitochondrial alleles.}, } @article {pmid28973572, year = {2017}, author = {Grau, T and Brandt, A and DeLeon, S and Meixner, MD and Strauß, JF and Joop, G and Telschow, A}, title = {A Comparison of Wolbachia Infection Frequencies in Varroa With Prevalence of Deformed Wing Virus.}, journal = {Journal of insect science (Online)}, volume = {17}, number = {3}, pages = {}, pmid = {28973572}, issn = {1536-2442}, mesh = {Animals ; Insect Viruses/*isolation & purification ; Varroidae/*virology ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia are widely distributed bacterial endosymbionts of arthropods and filarial nematodes. These bacteria can affect host fitness in a variety of ways, such as protecting hosts against viruses and other pathogens. Here, we investigate the possible role of Wolbachia in the prevalence of the deformed wing virus (DWV), a highly virulent pathogen of honey bees (Apis mellifera) that is transmitted by parasitic Varroa mites (Varroa destructor). About 180 Varroa mites from 18 beehives were tested for infection with Wolbachia and DWV. We first screened for Wolbachia using two standard primers (wsp and 16S rDNA), and found 26% of the mites to be positive for Wolbachia using the wsp primer and 64% of the mites to be positive using the 16S rDNA primer. Using these intermediate Wolbachia frequencies, we then tested for statistical correlations with virus infection frequencies. The analysis revealed a significant positive correlation between DWV and Wolbachia using the wsp primer, but no significant association between DWV and Wolbachia using the 16S rDNA primer. In conclusion, there is no evidence for an anti-pathogenic effect of Wolbachia in V. destructor, but weak evidence for a pro-pathogenic effect. These results encourage further examination of Wolbachia-virus interactions in Varroa mites since an increased vector competence of the mites may significantly impact disease outbreaks in honey bees.}, } @article {pmid28969836, year = {2017}, author = {Khowawisetsut, L and Sarasombath, PT and Thammapalo, S and Loymek, S and Korbarsa, T and Nochote, H and Phuakrod, A and Choochote, W and Wongkamchai, S}, title = {Therapeutic trial of doxycyclin plus ivermectin for the treatment of Brugia malayi naturally infected cats.}, journal = {Veterinary parasitology}, volume = {245}, number = {}, pages = {42-47}, doi = {10.1016/j.vetpar.2017.08.009}, pmid = {28969836}, issn = {1873-2550}, mesh = {Animals ; *Brugia malayi ; Cat Diseases/drug therapy/*parasitology ; Cats ; Doxycycline/administration & dosage/*therapeutic use ; Drug Therapy, Combination ; Filariasis/drug therapy/parasitology/*veterinary ; Ivermectin/administration & dosage/*therapeutic use ; }, abstract = {Lymphatic filariasis (LF) is one of the neglected tropical diseases which causes permanent and long term disability worldwide. LF is caused by filarial nematode parasites, i.e. Wuchereria bancrofti, Brugia malayi, and B. timori. All available antifilarial drugs currently being used have shown a limited adulticidal activity. Discoveries of endosymbiont rickettsia-like bacterium, Wolbachia in filarial nematodes provided a novel approach for antibiotic use in eradication of filarial diseases. The earlier studies revealed the macrofilaricidal efficacy of doxycycline against filarial nematodes. Chemotherapeutic efficiency of doxycycline has been studied against many filarial parasites, but there are still no therapeutic trials of the drug regimens for B. malayi treatment in naturally infected cats. Thus, this study would be the first attempt to study the efficiency of doxycycline (DOXY) alone or in combination with ivermectin (IVM) for treatment of B. malayi in naturally infected cats. A total of 26 B. malayi-infected cats in the endemic areas were recruited and divided into 3 groups, receiving different treatment regimens; a single dose of ivermectin only (IVM), doxycycline only (DOXY) and a combination of ivermectin and doxycycline (DOXY-IVM). The efficacy of each therapatic regimen was evaluated by detecting the presence of microfilaria using parasitological and molecular techniques monthly up to 2 years after starting the treatment. The IVM treated group had a significant rapid reduction of microfilariae in the first month; however, recurrence of microfilaraemia was observed in some cats. By contrast, the DOXY and DOXY-IVM groups showed a better result with a gradual decrease in microfilariae with no recurrence. These 2 groups were not only virtually deprived of infection but also sustained the sterility of infection through the course of study. These results revealed the advantages of using in B. malayi treatment in cats. Doxycycline showed to have both microfilaricidal and adulticidal effects on lymphatic filariae which maintained the long-term response to control of B. malayi infection in cats.}, } @article {pmid28959730, year = {2017}, author = {Johnston, KL and Cook, DAN and Berry, NG and David Hong, W and Clare, RH and Goddard, M and Ford, L and Nixon, GL and O'Neill, PM and Ward, SA and Taylor, MJ}, title = {Identification and prioritization of novel anti-Wolbachia chemotypes from screening a 10,000-compound diversity library.}, journal = {Science advances}, volume = {3}, number = {9}, pages = {eaao1551}, pmid = {28959730}, issn = {2375-2548}, mesh = {Anti-Bacterial Agents/*chemistry/*pharmacology ; Cluster Analysis ; Computational Biology/methods ; Drug Discovery/methods ; Drug Evaluation, Preclinical/*methods ; Humans ; Reproducibility of Results ; *Small Molecule Libraries ; Wolbachia/*drug effects ; Workflow ; }, abstract = {Lymphatic filariasis and onchocerciasis are two important neglected tropical diseases (NTDs) that cause severe disability. Control efforts are hindered by the lack of a safe macrofilaricidal drug. Targeting the Wolbachia bacterial endosymbionts in these parasites with doxycycline leads to a macrofilaricidal outcome, but protracted treatment regimens and contraindications restrict its widespread implementation. The Anti-Wolbachia consortium aims to develop improved anti-Wolbachia drugs to overcome these barriers. We describe the first screening of a large, diverse compound library against Wolbachia. This whole-organism screen, streamlined to reduce bottlenecks, produced a hit rate of 0.5%. Chemoinformatic analysis of the top 50 hits led to the identification of six structurally diverse chemotypes, the disclosure of which could offer interesting avenues of investigation to other researchers active in this field. An example of hit-to-lead optimization is described to further demonstrate the potential of developing these high-quality hit series as safe, efficacious, and selective anti-Wolbachia macrofilaricides.}, } @article {pmid28957585, year = {2017}, author = {Betelman, K and Caspi-Fluger, A and Shamir, M and Chiel, E}, title = {Identification and characterization of bacterial symbionts in three species of filth fly parasitoids.}, journal = {FEMS microbiology ecology}, volume = {93}, number = {9}, pages = {}, doi = {10.1093/femsec/fix107}, pmid = {28957585}, issn = {1574-6941}, mesh = {Animals ; Enterobacteriaceae/genetics/*isolation & purification ; Multilocus Sequence Typing ; Muscidae ; Phylogeny ; Pupa/*growth & development ; Reproduction ; Rickettsia/genetics/*isolation & purification ; Symbiosis/*physiology ; Wasps/*microbiology ; Weevils/*microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Facultative bacterial symbionts are widespread among insects and have diverse effects on their biology. Here, we focused on bacterial symbionts of three ecologically and economically important filth flies parasitoid species-Spalangia cameroni, Spalangia endius and Muscidifurax raptor. Both Spalangia species harbored a Sodalis bacterium that is closely related to Sodalis praecaptivus (a free-living bacterium) and to Sodalis symbionts of weevils. This is the only case of Sodalis infection in the important order Hymenoptera. We also found, for the first time in this parasitoid guild, a Rickettsia infecting the two Spalangia spp., albeit in much higher prevalence in S. cameroni. Molecular and phylogenetic analyses revealed that it is closely related to Rickettsia felis and other Rickettsia species from the 'transitional' group. All three parasitoid species harbored Wolbachia. Using multi-locus sequence typing, we found that M. raptor harbors a single Wolbachia strain whereas the Spalangia spp. have multiple strains. By controlled crossings, we found that Wolbachia infection in S. endius causes incomplete cytoplasmic incompatibility and increased longevity, thereby promoting Wolbachia's spread. In contrast, no effects of Wolbachia on the reproduction and longevity of M. raptor were found. This study underscores the diversity and nature of symbiotic interactions between microbes and insects.}, } @article {pmid28939989, year = {2017}, author = {Kaiser, L and Dupas, S and Branca, A and Herniou, EA and Clarke, CW and Capdevielle Dulac, C and Obonyo, J and Benoist, R and Gauthier, J and Calatayud, PA and Silvain, JF and Le Ru, BP}, title = {The Cotesia sesamiae story: insight into host-range evolution in a Hymenoptera parasitoid and implication for its use in biological control programs.}, journal = {Genetica}, volume = {145}, number = {6}, pages = {455-468}, pmid = {28939989}, issn = {1573-6857}, support = {ANR-12-ADAP-0001//Agence Nationale de la Recherche/ ; }, mesh = {Adaptation, Biological ; Animals ; *Biological Control Agents ; *Biological Evolution ; Genetic Speciation ; Host-Parasite Interactions ; Kenya ; Plants ; Wasps/*physiology ; }, abstract = {This review covers nearly 20 years of studies on the ecology, physiology and genetics of the Hymenoptera Cotesia sesamiae, an African parasitoid of Lepidoptera that reduces populations of common maize borers in East and South Africa. The first part of the review presents studies based on sampling of C. sesamiae from maize crops in Kenya. From this agrosystem including one host plant and three main host borer species, studies revealed two genetically differentiated populations of C. sesamiae species adapted to their local host community, and showed that their differentiation involved the joint evolution of virulence genes and sensory mechanisms of host acceptance, reinforced by reproductive incompatibility due to Wolbachia infection status and natural inbreeding. In the second part, we consider the larger ecosystem of wild Poales plant species hosting many Lepidoptera stem borer species that are potential hosts for C. sesamiae. The hypothesis of other host-adapted C. sesamiae populations was investigated based on a large sampling of stem borer larvae on various Poales across sub-Saharan Africa. The sampling provided information on the respective contribution of local hosts, biogeography and Wolbachia in the genetic structure of C. sesamiae populations. Molecular evolution analyses highlighted that several bracovirus genes were under positive selection, some of them being under different selection pressure in C. sesamiae populations adapted to different hosts. This suggests that C. sesamiae host races result from co-evolution acting at the local scale on different bracovirus genes. The third part considers the mechanisms driving specialization. C. sesamiae host races are more or less host-specialized. This character is crucial for efficient and environmentally-safe use of natural enemies for biological control of pests. One method to get an insight in the evolutionary stability of host-parasite associations is to characterize the phylogenetic relationships between the so-called host-races. Based on the construction of a phylogeny of C. sesamiae samples from various host- and plant species, we revealed three main lineages. Mechanisms of differentiation are discussed with regard to the geography and ecology of the samples. One of the lineage presented all the hallmarks of a distinct species, which has been morphologically described and is now studied in the perspective of being used as biological control agent against Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae), a major maize pest in West Africa and Mediterranean countries (see Benoist et al. 2017). The fourth part reviews past and present use of C. sesamiae in biological control, and points out the interest of such molecular ecology studies to reconcile biodiversity and food security stakes in future biological control.}, } @article {pmid28939962, year = {2018}, author = {Nadin, G and Strugarek, M and Vauchelet, N}, title = {Hindrances to bistable front propagation: application to Wolbachia invasion.}, journal = {Journal of mathematical biology}, volume = {76}, number = {6}, pages = {1489-1533}, pmid = {28939962}, issn = {1432-1416}, mesh = {Aedes/*microbiology/virology ; Animals ; Arbovirus Infections/prevention & control/transmission ; Arboviruses/pathogenicity ; Computational Biology ; Computer Simulation ; Dengue/prevention & control/transmission ; Host Microbial Interactions ; Humans ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/*microbiology/virology ; Pest Control, Biological/statistics & numerical data ; Population Density ; Wolbachia/*pathogenicity/physiology ; }, abstract = {We study the biological situation when an invading population propagates and replaces an existing population with different characteristics. For instance, this may occur in the presence of a vertically transmitted infection causing a cytoplasmic effect similar to the Allee effect (e.g. Wolbachia in Aedes mosquitoes): the invading dynamics we model is bistable. We aim at quantifying the propagules (what does it take for an invasion to start?) and the invasive power (how far can an invading front go, and what can stop it?). We rigorously show that a heterogeneous environment inducing a strong enough population gradient can stop an invading front, which will converge in this case to a stable front. We characterize the critical population jump, and also prove the existence of unstable fronts above the stable (blocking) fronts. Being above the maximal unstable front enables an invading front to clear the obstacle and propagate further. We are particularly interested in the case of artificial Wolbachia infection, used as a tool to fight arboviruses.}, } @article {pmid28939497, year = {2017}, author = {Soni, M and Bhattacharya, C and Sharma, J and Khan, SA and Dutta, P}, title = {Molecular typing and phylogeny of Wolbachia: A study from Assam, North-Eastern part of India.}, journal = {Acta tropica}, volume = {176}, number = {}, pages = {421-426}, doi = {10.1016/j.actatropica.2017.09.005}, pmid = {28939497}, issn = {1873-6254}, mesh = {Aedes/*microbiology ; Animals ; Anopheles/*microbiology ; Culex/*microbiology ; Drosophila melanogaster/*microbiology ; Female ; India ; Molecular Typing ; Phylogeny ; Wolbachia/*classification/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia are maternally inherited endosymbiotic alphaproteobacteria, infecting 40-75% of arthropod species. Knowledge on distribution of native strains infecting mosquito vectors from endemic regions is essential for successful implementation of vector control interventions utilizing potential strains of Wolbachia. Study identified various native strains of Wolbachia inhabiting different mosquito species from field and colonised conditions of Assam. The fly Drosophila melanogaster was also included in our study.

METHODS: Different mosquito species collected from field viz; Aedes albopictus, Aedes aegypti, Anopheles hyrcanus, Anopheles annularis, Culex vishnui, Toxorhynchites splendens, Armegeries obturbans and fly Drosophila melanogaster were included in the study. Anopheles stephensi and Culex quinquefasciatus were obtained from RMRC, Dibrugarh mosquito colony y for Wolbachia screening. DNA was extracted from these species, amplified using group specific wsp primers followed by sequencing and phylogenetic analysis.

RESULTS: Aedes albopictus from Dibrugarh, Tinsukia and Sivasagar district showed superinfection with A and B group of Wolbachia but, Aedes albopictus from Tezpur district presented infection with A group only. Our study reports for the first time natural infection of Wolbachia A and B group from colonised Anopheles stephensi mosquito but reported no infection from field collected Anopheles hyrcanus or Anopheles annularis. Similarly Armigeres obturbans and Culex vishnui presented infection with only B group of Wolbachia. Drosophila melanogaster showed superinfection with A and B group. Toxorhynchites splendens, Aedes aegypti and Culex quinquefasciatus reported no infection with Wolbachia.

CONCLUSION: To the best of our knowledge this is the first study on Wolbachia screening from Northeast part of India and also first report of natural Wolbachia infection from colonised Anopheles stephensi species. The current understanding on distribution of Wolbachia strains naturally present within insect species from this geographical region should aid future Wolbachia mediated vector control strategies.}, } @article {pmid28932814, year = {2017}, author = {Simhadri, RK and Fast, EM and Guo, R and Schultz, MJ and Vaisman, N and Ortiz, L and Bybee, J and Slatko, BE and Frydman, HM}, title = {The Gut Commensal Microbiome of Drosophila melanogaster Is Modified by the Endosymbiont Wolbachia.}, journal = {mSphere}, volume = {2}, number = {5}, pages = {}, pmid = {28932814}, issn = {2379-5042}, abstract = {Endosymbiotic Wolbachia bacteria and the gut microbiome have independently been shown to affect several aspects of insect biology, including reproduction, development, life span, stem cell activity, and resistance to human pathogens, in insect vectors. This work shows that Wolbachia bacteria, which reside mainly in the fly germline, affect the microbial species present in the fly gut in a lab-reared strain. Drosophila melanogaster hosts two main genera of commensal bacteria-Acetobacter and Lactobacillus. Wolbachia-infected flies have significantly reduced titers of Acetobacter. Sampling of the microbiome of axenic flies fed with equal proportions of both bacteria shows that the presence of Wolbachia bacteria is a significant determinant of the composition of the microbiome throughout fly development. However, this effect is host genotype dependent. To investigate the mechanism of microbiome modulation, the effect of Wolbachia bacteria on Imd and reactive oxygen species pathways, the main regulators of immune response in the fly gut, was measured. The presence of Wolbachia bacteria does not induce significant changes in the expression of the genes for the effector molecules in either pathway. Furthermore, microbiome modulation is not due to direct interaction between Wolbachia bacteria and gut microbes. Confocal analysis shows that Wolbachia bacteria are absent from the gut lumen. These results indicate that the mechanistic basis of the modulation of composition of the microbiome by Wolbachia bacteria is more complex than a direct bacterial interaction or the effect of Wolbachia bacteria on fly immunity. The findings reported here highlight the importance of considering the composition of the gut microbiome and host genetic background during Wolbachia-induced phenotypic studies and when formulating microbe-based disease vector control strategies. IMPORTANCEWolbachia bacteria are intracellular bacteria present in the microbiome of a large fraction of insects and parasitic nematodes. They can block mosquitos' ability to transmit several infectious disease-causing pathogens, including Zika, dengue, chikungunya, and West Nile viruses and malaria parasites. Certain extracellular bacteria present in the gut lumen of these insects can also block pathogen transmission. However, our understanding of interactions between Wolbachia and gut bacteria and how they influence each other is limited. Here we show that the presence of Wolbachia strain wMel changes the composition of gut commensal bacteria in the fruit fly. Our findings implicate interactions between bacterial species as a key factor in determining the overall composition of the microbiome and thus reveal new paradigms to consider in the development of disease control strategies.}, } @article {pmid28913925, year = {2018}, author = {Blackwood, JC and Vargas, R and Fauvergue, X}, title = {A cascade of destabilizations: Combining Wolbachia and Allee effects to eradicate insect pests.}, journal = {The Journal of animal ecology}, volume = {87}, number = {1}, pages = {59-72}, doi = {10.1111/1365-2656.12756}, pmid = {28913925}, issn = {1365-2656}, mesh = {Animals ; *Genetic Fitness ; Insecta/microbiology/*physiology ; Models, Biological ; *Pest Control, Biological ; Population Density ; Wolbachia/*physiology ; }, abstract = {The management of insect pests has long been dominated by the use of chemical insecticides, with the aim of instantaneously killing enough individuals to limit their damage. To minimize unwanted consequences, environmentally friendly approaches have been proposed that utilize biological control and take advantage of intrinsic demographic processes to reduce pest populations. We address the feasibility of a novel pest management strategy based on the release of insects infected with Wolbachia, which causes cytoplasmic incompatibilities in its host population, into a population with a pre-existing Allee effect. We hypothesize that the transient decline in population size caused by a successful invasion of Wolbachia can bring the population below its Allee threshold and, consequently, trigger extinction. We develop a stochastic population model that accounts for Wolbachia-induced cytoplasmic incompatibilities in addition to an Allee effect arising from mating failures at low population densities. Using our model, we identify conditions under which cytoplasmic incompatibilities and Allee effects successfully interact to drive insect pest populations towards extinction. Based on our results, we delineate control strategies based on introductions of Wolbachia-infected insects. We extend this analysis to evaluate control strategies that implement successive introductions of two incompatible Wolbachia strains. Additionally, we consider methods that combine Wolbachia invasion with mating disruption tactics to enhance the pre-existing Allee effect. We demonstrate that Wolbachia-induced cytoplasmic incompatibility and the Allee effect act independently from one another: the Allee effect does not modify the Wolbachia invasion threshold, and cytoplasmic incompatibilities only have a marginal effect on the Allee threshold. However, the interaction of these two processes can drive even large populations to extinction. The success of this method can be amplified by the introduction of multiple Wolbachia cytotypes as well as the addition of mating disruption. Our study extends the existing literature by proposing the use of Wolbachia introductions to capitalize on pre-existing Allee effects and consequently eradicate insect pests. More generally, it highlights the importance of transient dynamics, and the relevance of manipulating a cascade of destabilizatons for pest management.}, } @article {pmid28906572, year = {2018}, author = {Leggewie, M and Krumkamp, R and Badusche, M and Heitmann, A and Jansen, S and Schmidt-Chanasit, J and Tannich, E and Becker, SC}, title = {Culex torrentium mosquitoes from Germany are negative for Wolbachia.}, journal = {Medical and veterinary entomology}, volume = {32}, number = {1}, pages = {115-120}, doi = {10.1111/mve.12270}, pmid = {28906572}, issn = {1365-2915}, mesh = {Animals ; Culex/*microbiology ; Female ; Germany ; Species Specificity ; West Nile virus/*isolation & purification ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia (Rickettsiales: Anaplasmataceae) infects a wide range of arthropods, including several mosquito species. The bacterium is known to induce a plethora of phenotypes in its host, examples being the reproductive phenotype cytoplasmic incompatibility or resistance against infection with arboviruses. The latter is especially relevant when assessing the vector competence of mosquito species for emerging arboviruses. Thus, knowledge of Wolbachia infection status is important for the assessment of vector competence. To facilitate Wolbachia screening in mosquito populations, a quantitative polymerase chain reaction (qPCR) assay was developed to enable high-throughput analysis of mosquito samples. Using this assay, the Wolbachia infection status of the two most common Culex mosquito species in Germany, Culex pipiens biotype pipiens Linnaeus (Diptera: Culicidae) and Culex torrentium Martini (Diptera: Culicidae), was assessed. About 93% of all tested C. pipiens biotype pipiens individuals were positive for Wolbachia, whereas none of the C. torrentium samples was found to be infected. Furthermore, other applications of the qPCR assay were explored by assessing a potential link between the levels of Wolbachia and West Nile virus (WNV) infections in German C. pipiens biotype pipiens mosquitoes. No relationship was found between the two variables, indicating that a Wolbachia-induced antiviral phenotype in this mosquito population is not exclusively attributable to the general level of bacterial infection.}, } @article {pmid28905230, year = {2017}, author = {Huber, D and Reil, I and Duvnjak, S and Jurković, D and Lukačević, D and Pilat, M and Beck, A and Mihaljević, Ž and Vojta, L and Polkinghorne, A and Beck, R}, title = {Molecular detection of Anaplasma platys, Anaplasma phagocytophilum and Wolbachia sp. but not Ehrlichia canis in Croatian dogs.}, journal = {Parasitology research}, volume = {116}, number = {11}, pages = {3019-3026}, pmid = {28905230}, issn = {1432-1955}, mesh = {Anaplasma/classification/*isolation & purification ; Anaplasma phagocytophilum/genetics/*isolation & purification ; Anaplasmosis/epidemiology/*microbiology ; Animals ; Base Sequence ; Croatia/epidemiology ; Dog Diseases/epidemiology/*microbiology ; Dogs ; Ehrlichia canis/*isolation & purification ; Ehrlichiosis/veterinary ; Phylogeny ; Polymerase Chain Reaction/veterinary ; RNA, Bacterial ; RNA, Ribosomal, 16S/genetics ; Rhipicephalus sanguineus/microbiology ; Rickettsiaceae Infections/epidemiology/microbiology/*veterinary ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceae-specific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. platys in 2.5% (CI 1.6-3.4%, 27 dogs), A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.}, } @article {pmid28904344, year = {2017}, author = {Geoghegan, V and Stainton, K and Rainey, SM and Ant, TH and Dowle, AA and Larson, T and Hester, S and Charles, PD and Thomas, B and Sinkins, SP}, title = {Perturbed cholesterol and vesicular trafficking associated with dengue blocking in Wolbachia-infected Aedes aegypti cells.}, journal = {Nature communications}, volume = {8}, number = {1}, pages = {526}, pmid = {28904344}, issn = {2041-1723}, support = {/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_PC_15087/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/*metabolism/*microbiology/virology ; Animals ; Cholesterol/*metabolism ; Cytoplasmic Vesicles/*metabolism ; Dengue/*transmission/virology ; Dengue Virus/*physiology ; Humans ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular maternally inherited bacteria that can spread through insect populations and block virus transmission by mosquitoes, providing an important approach to dengue control. To better understand the mechanisms of virus inhibition, we here perform proteomic quantification of the effects of Wolbachia in Aedes aegypti mosquito cells and midgut. Perturbations are observed in vesicular trafficking, lipid metabolism and in the endoplasmic reticulum that could impact viral entry and replication. Wolbachia-infected cells display a differential cholesterol profile, including elevated levels of esterified cholesterol, that is consistent with perturbed intracellular cholesterol trafficking. Cyclodextrins have been shown to reverse lipid accumulation defects in cells with disrupted cholesterol homeostasis. Treatment of Wolbachia-infected Ae. aegypti cells with 2-hydroxypropyl-β-cyclodextrin restores dengue replication in Wolbachia-carrying cells, suggesting dengue is inhibited in Wolbachia-infected cells by localised cholesterol accumulation. These results demonstrate parallels between the cellular Wolbachia viral inhibition phenotype and lipid storage genetic disorders. Wolbachia infection of mosquitoes can block dengue virus infection and is tested in field trials, but the mechanism of action is unclear. Using proteomics, Geoghegan et al. here identify effects of Wolbachia on cholesterol homeostasis and dengue virus replication in Aedes aegypti.}, } @article {pmid28902190, year = {2017}, author = {Lindsey, ARI and Stouthamer, R}, title = {The effects of outbreeding on a parasitoid wasp fixed for infection with a parthenogenesis-inducing Wolbachia symbiont.}, journal = {Heredity}, volume = {119}, number = {6}, pages = {411-417}, pmid = {28902190}, issn = {1365-2540}, mesh = {Animals ; Biological Coevolution ; Crosses, Genetic ; Female ; Fertility ; Heterozygote ; Homozygote ; Male ; Models, Genetic ; *Parthenogenesis ; *Sex Ratio ; Symbiosis ; Wasps/*genetics/microbiology ; *Wolbachia ; }, abstract = {Trichogramma wasps can be rendered asexual by infection with the maternally inherited symbiont Wolbachia. Previous studies indicate the Wolbachia strains infecting Trichogramma wasps are host-specific, inferred by failed horizontal transfer of Wolbachia to novel Trichogramma hosts. Additionally, Trichogramma can become dependent upon their Wolbachia infection for the production of female offspring, leaving them irreversibly asexual, further linking host and symbiont. We hypothesized Wolbachia strains infecting irreversibly asexual, resistant to horizontal transfer Trichogramma would show adaptation to a particular host genetic background. To test this, we mated Wolbachia-dependent females with males from a Wolbachia-naïve population to create heterozygous wasps. We measured sex ratios and fecundity, a proxy for Wolbachia fitness, produced by heterozygous wasps, and by their recombinant offspring. We find a heterozygote advantage, resulting in higher fitness for Wolbachia, as wasps will produce more offspring without any reduction in the proportion of females. While recombinant wasps did not differ in total fecundity after 10 days, recombinants produced fewer offspring early on, leading to an increased female-biased sex ratio for the whole brood. Despite the previously identified barriers to horizontal transfer of Wolbachia to and from Trichogramma pretiosum, there were no apparent barriers for Wolbachia to induce parthenogenesis in these non-native backgrounds. This is likely due to the route of infection being introgression rather than horizontal transfer, and possibly the co-evolution of Wolbachia with the mitochondria rather than the nuclear genome. These results help to elucidate the mechanisms by which Wolbachia adapt to hosts and the evolution of host-symbiont phenotypes.}, } @article {pmid28892518, year = {2017}, author = {Rohrscheib, CE and Frentiu, FD and Horn, E and Ritchie, FK and van Swinderen, B and Weible, MW and O'Neill, SL and Brownlie, JC}, title = {Response to: Comment on Rohrscheib et al. 2016 "Intensity of mutualism breakdown is determined by temperature not amplification of Wolbachia genes".}, journal = {PLoS pathogens}, volume = {13}, number = {9}, pages = {e1006521}, pmid = {28892518}, issn = {1553-7374}, mesh = {Genome, Bacterial ; Phylogeny ; *Symbiosis ; Temperature ; Wolbachia/*genetics ; }, } @article {pmid28892498, year = {2017}, author = {Chrostek, E and Teixeira, L}, title = {Comment on Rohrscheib et al. 2016 "Intensity of mutualism breakdown is determined by temperature not amplification of Wolbachia genes".}, journal = {PLoS pathogens}, volume = {13}, number = {9}, pages = {e1006540}, pmid = {28892498}, issn = {1553-7374}, } @article {pmid28878066, year = {2017}, author = {Gebiola, M and Giorgini, M and Kelly, SE and Doremus, MR and Ferree, PM and Hunter, MS}, title = {Cytological analysis of cytoplasmic incompatibility induced by Cardinium suggests convergent evolution with its distant cousin Wolbachia.}, journal = {Proceedings. Biological sciences}, volume = {284}, number = {1862}, pages = {}, pmid = {28878066}, issn = {1471-2954}, mesh = {Aneuploidy ; Animals ; *Cytophagaceae ; Cytoplasm/*microbiology ; Female ; Male ; Mitosis ; Reproduction ; Symbiosis ; Wasps/*microbiology ; Wolbachia ; }, abstract = {Cytoplasmic incompatibility (CI) is a conditional sterility in numerous arthropods that is caused by inherited, intracellular bacteria such as Wolbachia Matings between males carrying CI-inducing Wolbachia and uninfected females, or between males and females infected with different Wolbachia strains, result in progeny that die during very early embryogenesis. Multiple studies in diploid (Drosophila) and haplodiploid (Nasonia) insects have shown that CI-Wolbachia cause a failure of the paternally derived chromatin from resolving into distinct chromosomes. This leads to the formation of chromatin bridges and other mitotic defects as early as the first mitotic division, and to early mitotic arrest. It is currently unknown if CI-inducing symbionts other than Wolbachia affect similar cellular processes. Here, we investigated CI caused by an unrelated bacterium, Cardinium, which naturally infects a parasitic wasp, Encarsia suzannae CI crosses in this host-symbiont system resulted in early mitotic defects including asynchrony of paternal and maternal chromosome sets as they enter mitosis, chromatin bridges and improper chromosome segregation that spanned across multiple mitotic divisions, triggering embryonic death through accumulated aneuploidy. We highlight small differences with CI-Wolbachia, which could be due to the underlying CI mechanism or host-specific effects. Our results suggest a convergence of CI-related cellular phenotypes between these two unrelated symbionts.}, } @article {pmid28856446, year = {2018}, author = {Bliman, PA and Aronna, MS and Coelho, FC and da Silva, MAHB}, title = {Ensuring successful introduction of Wolbachia in natural populations of Aedes aegypti by means of feedback control.}, journal = {Journal of mathematical biology}, volume = {76}, number = {5}, pages = {1269-1300}, pmid = {28856446}, issn = {1432-1416}, mesh = {Aedes/*microbiology ; Animals ; Arbovirus Infections/prevention & control/transmission ; Computer Simulation ; Dengue/*prevention & control/transmission ; Female ; Humans ; Male ; Mathematical Concepts ; Models, Biological ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods/statistics & numerical data ; Wolbachia/pathogenicity/*physiology ; }, abstract = {The control of the spread of dengue fever by introduction of the intracellular parasitic bacterium Wolbachia in populations of the vector Aedes aegypti, is presently one of the most promising tools for eliminating dengue, in the absence of an efficient vaccine. The success of this operation requires locally careful planning to determine the adequate number of individuals carrying the Wolbachia parasite that need to be introduced into the natural population. The introduced mosquitoes are expected to eventually replace the Wolbachia-free population and guarantee permanent protection against the transmission of dengue to human. In this study, we propose and analyze a model describing the fundamental aspects of the competition between mosquitoes carrying Wolbachia and mosquitoes free of the parasite. We then use feedback control techniques to devise an introduction protocol that is proved to guarantee that the population converges to a stable equilibrium where the totality of mosquitoes carry Wolbachia.}, } @article {pmid28855327, year = {2017}, author = {Sullivan, W}, title = {Wolbachia, bottled water, and the dark side of symbiosis.}, journal = {Molecular biology of the cell}, volume = {28}, number = {18}, pages = {2343-2346}, pmid = {28855327}, issn = {1939-4586}, mesh = {Animals ; Humans ; Symbiosis/*physiology ; }, abstract = {Obligate endosymbiosis is operationally defined when loss or removal of the endosymbiont from the host results in the death of both. Whereas these relationships are typically viewed as mutualistic, molecular and cellular analysis reveals numerous instances in which these symbiotic relationships are established by alternative, nonmutualistic strategies. The endosymbiont usurps or integrates into core host processes, creating a need where none previously existed. Here I discuss examples of these addictive symbiotic relationships and how they are a likely outcome of all complex evolving systems.}, } @article {pmid28854601, year = {2017}, author = {Rice, DW and Sheehan, KB and Newton, ILG}, title = {Large-Scale Identification of Wolbachia pipientis Effectors.}, journal = {Genome biology and evolution}, volume = {9}, number = {7}, pages = {1925-1937}, pmid = {28854601}, issn = {1759-6653}, mesh = {Animals ; Bacterial Proteins/genetics/*metabolism ; Computational Biology/*methods ; Drosophila melanogaster/genetics/metabolism/*microbiology ; Gene Expression Regulation, Bacterial ; Genetic Association Studies ; Host-Pathogen Interactions ; Phylogeny ; Saccharomyces cerevisiae/genetics/growth & development ; Symbiosis ; Wolbachia/genetics/*metabolism ; }, abstract = {Wolbachia pipientis is an intracellular symbiont of arthropods well known for the reproductive manipulations induced in the host and, more recently, for the ability of Wolbachia to block virus replication in insect vectors. Since Wolbachia cannot yet be genetically manipulated, and due to the constraints imposed when working with an intracellular symbiont, little is known about mechanisms used by Wolbachia for host interaction. Here we employed a bioinformatics pipeline and identified 163 candidate effectors, potentially secreted by Wolbachia into the host cell. A total of 84 of these candidates were then subjected to a screen of growth defects induced in yeast upon heterologous expression which identified 14 top candidates likely secreted by Wolbachia. These predicted secreted effectors may function in concert as we find that their native expression is correlated and is highly upregulated at specific time points during Drosophila development. In addition, the evolutionary histories of some of these predicted effectors are also correlated, suggesting they may function together, or in the same pathway, during host infection. Similarly, most of these predicted effectors are limited to one or two Wolbachia strains-perhaps reflecting shared evolutionary history and strain specific functions in host manipulation. Identification of these Wolbachia candidate effectors is the first step in dissecting the mechanisms of symbiont-host interaction in this important system.}, } @article {pmid28845255, year = {2017}, author = {Vi, TT and Kien, DT and Wills, B and Simmons, CP}, title = {A29 Wolbachia for dengue control; will dengue viruses evolve resistance?.}, journal = {Virus evolution}, volume = {3}, number = {Suppl 1}, pages = {}, doi = {10.1093/ve/vew036.028}, pmid = {28845255}, issn = {2057-1577}, } @article {pmid28842593, year = {2017}, author = {Ojha, A and Sinha, DK and Padmakumari, AP and Bentur, JS and Nair, S}, title = {Bacterial Community Structure in the Asian Rice Gall Midge Reveals a Varied Microbiome Rich in Proteobacteria.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {9424}, pmid = {28842593}, issn = {2045-2322}, mesh = {Animals ; *Biodiversity ; Diptera/*microbiology ; Metagenome ; Metagenomics/methods ; *Microbiota ; Proteobacteria/*classification/*genetics ; RNA, Ribosomal, 16S/genetics ; }, abstract = {The Asian rice gall midge (ARGM) has emerged as a model gall forming pest of rice. The ARGM infestation of rice results in failure of panicle formation and economic loss. Understanding the molecular basis of ARGM-rice interactions is very crucial in order to control this devastating pest of rice. The current investigation was devised to identify bacterial communities present in the ARGM and in addition the bacterial diversity in the maggots during their interaction with susceptible or resistant rice varieties. Sequencing of 16S rRNA bacterial gene (V3-V4 region) revealed differences in the microflora of the ARGM maggots feeding on susceptible or resistant rice hosts. Results revealed that Wolbachia was the predominant bacterium in pupae and adults while Pseudomonas was predominant in maggots. Further, we observed that members of proteobacteria were predominant across all the samples. There was high species diversity in maggots isolated from susceptible rice and a high representation of unclassified bacteria in maggots isolated from resistant rice. This is the first study that reports variation of microbiome of the ARGM, based on host phenotype from which it was isolated, and results suggest that these variations could have an important role in host's susceptibility.}, } @article {pmid28841917, year = {2017}, author = {Almeida, F and Suesdek, L}, title = {Effects of Wolbachia on ovarian apoptosis in Culex quinquefasciatus (Say, 1823) during the previtellogenic and vitellogenic periods.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {398}, pmid = {28841917}, issn = {1756-3305}, mesh = {Animals ; *Apoptosis ; Culex/*microbiology/*physiology ; Female ; Microscopy, Confocal ; Oocytes/microbiology/pathology ; Oogenesis ; Ovary/cytology/microbiology/ultrastructure ; Vitellogenesis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Apoptosis is programmed cell death that ordinarily occurs in ovarian follicular cells in various organisms. In the best-studied holometabolous insect, Drosophila, this kind of cell death occurs in all three cell types found in the follicles, sometimes leading to follicular atresia and egg degeneration. On the other hand, egg development, quantity and viability in the mosquito Culex quinquefasciatus are disturbed by the infection with the endosymbiont Wolbachia. Considering that Wolbachia alters reproductive traits, we hypothesised that such infection would also alter the apoptosis in the ovarian cells of this mosquito. The goal of this study was to comparatively describe the occurrence of apoptosis in Wolbachia-infected and uninfected ovaries of Cx. quinquefasciatus during oogenesis and vitellogenesis. For this, we recorded under confocal microscopy the occurrence of apoptosis in all three cell types of the ovarian follicle. In the first five days of adult life we observed oogenesis and, after a blood meal, the initiation step of vitellogenesis.

RESULTS: Apoptoses in follicular cells were found at all observation times during both oogenesis and vitellogenesis, and less commonly in nurse cells and the oocyte, as well as in atretic follicles. Our results suggested that apoptosis in follicular cells occurred in greater numbers in infected mosquitoes than in uninfected ones during the second and third days of adult life and at the initiation step of vitellogenesis.

CONCLUSIONS: The presence of Wolbachia leads to an increase of apoptosis occurrence in the ovaries of Cx. quinquefasciatus. Future studies should investigate if this augmented apoptosis frequency is the cause of the reduction in the number of eggs laid by Wolbachia-infected females. Follicular atresia is first reported in the previtellogenic period of oogenesis. Our findings may have implications for the use of Wolbachia as a mosquito and pathogens control strategy.}, } @article {pmid28835430, year = {2017}, author = {Panda, DK and Mohapatra, DP and Mohapatra, MM}, title = {Filarial breast lump.}, journal = {BMJ case reports}, volume = {2017}, number = {}, pages = {}, pmid = {28835430}, issn = {1757-790X}, mesh = {Adult ; Breast Diseases/*diagnosis/diagnostic imaging/drug therapy/surgery ; Diagnosis, Differential ; Diethylcarbamazine/administration & dosage/therapeutic use ; Elephantiasis, Filarial/*diagnosis/diagnostic imaging/drug therapy/surgery ; Female ; Filaricides/administration & dosage/therapeutic use ; Humans ; Mammography ; Mastectomy, Segmental ; Pain/etiology ; Wolbachia/isolation & purification ; }, abstract = {Lymphatic filariasis is one of the most debilitating and disfiguring scourges among all diseases. This report presents a case of a woman with recurrent breast nodularity after being previously operated for a suspected breast neoplasm. We would like to highlight the issue of similar clinical presentation of a filarial breast lump and other breast lesions leading to inappropriate therapy.}, } @article {pmid28829414, year = {2017}, author = {Ross, PA and Axford, JK and Richardson, KM and Endersby-Harshman, NM and Hoffmann, AA}, title = {Maintaining Aedes aegypti Mosquitoes Infected with Wolbachia.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {126}, pages = {}, pmid = {28829414}, issn = {1940-087X}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/physiology ; Animals ; Female ; Larva ; Male ; Oviposition ; *Wolbachia ; }, abstract = {Aedes aegypti mosquitoes experimentally infected with Wolbachia are being utilized in programs to control the spread of arboviruses such as dengue, chikungunya and Zika. Wolbachia-infected mosquitoes can be released into the field to either reduce population sizes through incompatible matings or to transform populations with mosquitoes that are refractory to virus transmission. For these strategies to succeed, the mosquitoes released into the field from the laboratory must be competitive with native mosquitoes. However, maintaining mosquitoes in the laboratory can result in inbreeding, genetic drift and laboratory adaptation which can reduce their fitness in the field and may confound the results of experiments. To test the suitability of different Wolbachia infections for deployment in the field, it is necessary to maintain mosquitoes in a controlled laboratory environment across multiple generations. We describe a simple protocol for maintaining Ae. aegypti mosquitoes in the laboratory, which is suitable for both Wolbachia-infected and wild-type mosquitoes. The methods minimize laboratory adaptation and implement outcrossing to increase the relevance of experiments to field mosquitoes. Additionally, colonies are maintained under optimal conditions to maximize their fitness for open field releases.}, } @article {pmid28824885, year = {2017}, author = {Wilmes, M and Meier, K and Schiefer, A and Josten, M and Otten, CF and Klöckner, A and Henrichfreise, B and Vollmer, W and Hoerauf, A and Pfarr, K}, title = {AmiD Is a Novel Peptidoglycan Amidase in Wolbachia Endosymbionts of Drosophila melanogaster.}, journal = {Frontiers in cellular and infection microbiology}, volume = {7}, number = {}, pages = {353}, pmid = {28824885}, issn = {2235-2988}, support = {MR/N501840/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Amidohydrolases/genetics/immunology/*metabolism ; Amino Acid Sequence ; Animals ; Arthropods/microbiology ; Cell Wall/metabolism ; Drosophila melanogaster/*microbiology ; Genetic Vectors ; Mutagenesis, Site-Directed ; Nematoda/microbiology ; Peptidoglycan/*biosynthesis/immunology ; Sequence Analysis, Protein ; Symbiosis ; Uridine Diphosphate N-Acetylmuramic Acid/analogs & derivatives/metabolism ; Wolbachia/*enzymology/genetics ; }, abstract = {Wolbachia endobacteria are obligate intracellular bacteria with a highly reduced genome infecting many arthropod and filarial species, in which they manipulate arthropod reproduction to increase their transmission and are essential for nematode development and survival. The Wolbachia genome encodes all enzymes required for the synthesis of the cell wall building block lipid II, although a peptidoglycan-like structure has not been detected. Despite the ability to synthesize lipid II, Wolbachia from arthropods and nematodes have only a subset of genes encoding enzymes involved in the periplasmic processing of lipid II and peptidoglycan recycling, with arthropods having two more than nematodes. We functionally analyzed the activity of the putative cell wall hydrolase AmiD from the Wolbachia endosymbiont of Drosophila melanogaster, an enzyme not encoded by the nematode endobacteria. Wolbachia AmiD has Zn[2+]-dependent amidase activity and cleaves intact peptidoglycan, monomeric lipid II and anhydromuropeptides, substrates that are generated during bacterial growth. AmiD may have been maintained in arthropod Wolbachia to avoid host immune recognition by degrading cell wall fragments in the periplasm. This is the first description of a wolbachial lipid II processing enzyme putatively expressed in the periplasm.}, } @article {pmid28824591, year = {2017}, author = {Jacob, F and Melachio, TT and Njitchouang, GR and Gimonneau, G and Njiokou, F and Abate, L and Christen, R and Reveillaud, J and Geiger, A}, title = {Intestinal Bacterial Communities of Trypanosome-Infected and Uninfected Glossina palpalis palpalis from Three Human African Trypanomiasis Foci in Cameroon.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {1464}, pmid = {28824591}, issn = {1664-302X}, abstract = {Glossina sp. the tsetse fly that transmits trypanosomes causing the Human or the Animal African Trypanosomiasis (HAT or AAT) can harbor symbiotic bacteria that are known to play a crucial role in the fly's vector competence. We hypothesized that other bacteria could be present, and that some of them could also influence the fly's vector competence. In this context the objectives of our work were: (a) to characterize the bacteria that compose the G. palpalis palpalis midgut bacteriome, (b) to evidence possible bacterial community differences between trypanosome-infected and non-infected fly individuals from a given AAT and HAT focus or from different foci using barcoded Illumina sequencing of the hypervariable V3-V4 region of the 16S rRNA gene. Forty G. p. palpalis flies, either infected by Trypanosoma congolense or uninfected were sampled from three trypanosomiasis foci in Cameroon. A total of 143 OTUs were detected in the midgut samples. Most taxa were identified at the genus level, nearly 50% at the species level; they belonged to 83 genera principally within the phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. Prominent representatives included Wigglesworthia (the fly's obligate symbiont), Serratia, and Enterobacter hormaechei. Wolbachia was identified for the first time in G. p. palpalis. The average number of bacterial species per tsetse sample was not significantly different regarding the fly infection status, and the hierarchical analysis based on the differences in bacterial community structure did not provide a clear clustering between infected and non-infected flies. Finally, the most important result was the evidence of the overall very large diversity of intestinal bacteria which, except for Wigglesworthia, were unevenly distributed over the sampled flies regardless of their geographic origin and their trypanosome infection status.}, } @article {pmid28821226, year = {2017}, author = {Saucereau, Y and Valiente Moro, C and Dieryckx, C and Dupuy, JW and Tran, FH and Girard, V and Potier, P and Mavingui, P}, title = {Comprehensive proteome profiling in Aedes albopictus to decipher Wolbachia-arbovirus interference phenomenon.}, journal = {BMC genomics}, volume = {18}, number = {1}, pages = {635}, pmid = {28821226}, issn = {1471-2164}, mesh = {Aedes/*metabolism/microbiology/virology ; Animals ; Arboviruses/*physiology ; Cell Line ; *Proteomics ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Aedes albopictus is a vector of arboviruses that cause severe diseases in humans such as Chikungunya, Dengue and Zika fevers. The vector competence of Ae. albopictus varies depending on the mosquito population involved and the virus transmitted. Wolbachia infection status in believed to be among key elements that determine viral transmission efficiency. Little is known about the cellular functions mobilized in Ae. albopictus during co-infection by Wolbachia and a given arbovirus. To decipher this tripartite interaction at the molecular level, we performed a proteome analysis in Ae. albopictus C6/36 cells mono-infected by Wolbachia wAlbB strain or Chikungunya virus (CHIKV), and bi-infected.

RESULTS: We first confirmed significant inhibition of CHIKV by Wolbachia. Using two-dimensional gel electrophoresis followed by nano liquid chromatography coupled with tandem mass spectrometry, we identified 600 unique differentially expressed proteins mostly related to glycolysis, translation and protein metabolism. Wolbachia infection had greater impact on cellular functions than CHIKV infection, inducing either up or down-regulation of proteins associated with metabolic processes such as glycolysis and ATP metabolism, or structural glycoproteins and capsid proteins in the case of bi-infection with CHIKV. CHIKV infection inhibited expression of proteins linked with the processes of transcription, translation, lipid storage and miRNA pathways.

CONCLUSIONS: The results of our proteome profiling have provided new insights into the molecular pathways involved in tripartite Ae. albopictus-Wolbachia-CHIKV interaction and may help defining targets for the better implementation of Wolbachia-based strategies for disease transmission control.}, } @article {pmid28807085, year = {2018}, author = {Karimi, S and Izadi, H and Askari Seyahooei, M and Bagheri, A and Khodaygan, P}, title = {Variation in bacterial endosymbionts associated with the date palm hopper, Ommatissus lybicus populations.}, journal = {Bulletin of entomological research}, volume = {108}, number = {2}, pages = {271-281}, doi = {10.1017/S0007485317000633}, pmid = {28807085}, issn = {1475-2670}, mesh = {Animals ; Hemiptera/*microbiology ; Phylogeny ; Symbiosis ; }, abstract = {The date palm hopper, Ommatissus lybicus, is a key pest of the date palm, which is expected to be comprised of many allopatric populations. The current study was carried out to determine bacterial endosymbiont diversity in the different populations of this pest. Ten date palm hopper populations were collected from the main date palm growing regions in Iran and an additional four samples from Pakistan, Oman, Egypt and Tunisia for detection of primary and secondary endosymbionts using polymerase chain reaction (PCR) assay with their specific primers. The PCR products were directly sequenced and edited using SeqMan software. The consensus sequences were subjected to a BLAST similarity search. The results revealed the presence of 'Candidatus Sulcia muelleri' (primary endosymbiont) and Wolbachia, Arsenophonus and Enterobacter (secondary endosymbionts) in all populations. This assay failed to detect 'Candidatus Nasuia deltocephalinicola' and Serratia in these populations. 'Ca. S. muelleri' exhibited a 100% infection frequency in populations and Wolbachia, Arsenophonus and Enterobacter demonstrated 100, 93.04 and 97.39% infection frequencies, respectively. The infection rate of Arsenophonus and Enterobacter ranged from 75 to 100% and 62.5 to 100%, respectively, in different populations of the insect. The results demonstrated multiple infections by 'Ca. Sulcia muelleri', Wolbachia, Arsenophonus and Enterobacter in the populations and may suggest significant roles for these endosymbionts on date palm hopper population fitness. This study provides an insight to endosymbiont variation in the date palm hopper populations; however, further investigation is needed to examine how these endosymbionts may affect host fitness.}, } @article {pmid28805637, year = {2017}, author = {Terradas, G and McGraw, EA}, title = {Wolbachia-mediated virus blocking in the mosquito vector Aedes aegypti.}, journal = {Current opinion in insect science}, volume = {22}, number = {}, pages = {37-44}, doi = {10.1016/j.cois.2017.05.005}, pmid = {28805637}, issn = {2214-5753}, mesh = {Aedes/microbiology/*virology ; Animals ; Arbovirus Infections/prevention & control/transmission ; Arboviruses/physiology ; Host Microbial Interactions ; Immunity, Innate ; Mosquito Vectors/immunology/microbiology/virology ; Pest Control, Biological/methods ; Wolbachia/*physiology ; }, abstract = {Viruses transmitted by mosquitoes such as dengue, Zika and West Nile cause a threat to global health due to increased geographical range and frequency of outbreaks. The bacterium Wolbachia pipientis may be the solution reducing disease transmission. Though commonly missing in vector species, the bacterium was artificially and stably introduced into Aedes aegypti to assess its potential for biocontrol. When infected with Wolbachia, mosquitoes become refractory to infection by a range of pathogens, including the aforementioned viruses. How the bacterium is conferring this phenotype remains unknown. Here we discuss current hypotheses in the field for the mechanistic basis of pathogen blocking and evaluate the evidence from mosquitoes and related insects.}, } @article {pmid28805302, year = {2017}, author = {Pilgrim, J and Ander, M and Garros, C and Baylis, M and Hurst, GDD and Siozios, S}, title = {Torix group Rickettsia are widespread in Culicoides biting midges (Diptera: Ceratopogonidae), reach high frequency and carry unique genomic features.}, journal = {Environmental microbiology}, volume = {19}, number = {10}, pages = {4238-4255}, pmid = {28805302}, issn = {1462-2920}, support = {BBS/E/I/00001701/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BBS/E/I/00007036/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BBS/E/I/00007038/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BBS/E/I/00007039/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Base Sequence ; Cell Wall/metabolism ; Ceratopogonidae/*microbiology ; Female ; Genome, Bacterial/*genetics ; Genomics ; Insect Vectors/*microbiology ; Lipopolysaccharides/genetics ; Pentose Phosphate Pathway/genetics ; Phylogeny ; Rickettsia/*classification/*genetics/isolation & purification ; Sequence Analysis, DNA ; Symbiosis/genetics ; }, abstract = {There is increasing interest in the heritable bacteria of invertebrate vectors of disease as they present novel targets for control initiatives. Previous studies on biting midges (Culicoides spp.), known to transmit several RNA viruses of veterinary importance, have revealed infections with the endosymbiotic bacteria, Wolbachia and Cardinium. However, rickettsial symbionts in these vectors are underexplored. Here, we present the genome of a previously uncharacterized Rickettsia endosymbiont from Culicoides newsteadi (RiCNE). This genome presents unique features potentially associated with host invasion and adaptation, including genes for the complete non-oxidative phase of the pentose phosphate pathway, and others predicted to mediate lipopolysaccharides and cell wall modification. Screening of 414 Culicoides individuals from 29 Palearctic or Afrotropical species revealed that Rickettsia represent a widespread but previously overlooked association, reaching high frequencies in midge populations and present in 38% of the species tested. Sequence typing clusters the Rickettsia within the Torix group of the genus, a group known to infect several aquatic and hematophagous taxa. FISH analysis indicated the presence of Rickettsia bacteria in ovary tissue, indicating their maternal inheritance. Given the importance of biting midges as vectors, a key area of future research is to establish the impact of this endosymbiont on vector competence.}, } @article {pmid28803406, year = {2018}, author = {Rahimi-Kaldeh, S and Ashouri, A and Bandani, A}, title = {Does Wolbachia Infection Change the Overwintering Ability of Trichogramma brassicae (Hymenoptera: Trichogrammatidae)?.}, journal = {Neotropical entomology}, volume = {47}, number = {5}, pages = {583-590}, pmid = {28803406}, issn = {1678-8052}, mesh = {Animals ; Iran ; Lepidoptera/parasitology ; Ovum/parasitology ; *Seasons ; Wasps/*microbiology/*physiology ; *Wolbachia ; }, abstract = {The overwintering ability of a biological control agent helps predict potential survival and establishment in natural conditions. Here, we compared the overwintering ability in sexual and asexual Trichogramma brassicae Bezdenko, a key natural enemy of Lepidopteran pests in Iran over two consecutive winters in central Alborz Mountains. Our aim was to investigate the effect of Wolbachia infection on the overwintering rate of T. brassicae after emergence in the subsequent year. We also wanted to know whether there is any difference in the overwintering capacity of T. brassicae populations either in terms of exposure date or emergence date. Parasitized host eggs were exposed under outdoor conditions between 8 October and 27 November 2013 and between 8 October and 11 December 2014. Samples were checked weekly until adult emergence of overwintered individuals in the following spring, and proportion of adult emergence was recorded on each exposure date. Our results demonstrated that both populations were able to overwinter in natural conditions of central Alborz Mountains; however, sexual T. brassicae was more successful in overwintering than asexual one. No differences were observed in the emergence date and maximum emergence rate between two populations. Our study clearly identified a potential negative impact of Wolbachia on fitness of T. brassicae by a lower rate of emergence.}, } @article {pmid28799248, year = {2018}, author = {DA Rocha, NO and Lambert, SM and Dias-Lima, AG and Julião, FS and Souza, BMPS}, title = {Molecular detection of Wolbachia pipientis in natural populations of sandfly vectors of Leishmania infantum in endemic areas: first detection in Lutzomyia longipalpis.}, journal = {Medical and veterinary entomology}, volume = {32}, number = {1}, pages = {111-114}, doi = {10.1111/mve.12255}, pmid = {28799248}, issn = {1365-2915}, mesh = {Animals ; Brazil ; Communicable Disease Control/methods ; Female ; Haplotypes ; Insect Vectors/*microbiology ; Leishmania infantum/physiology ; Leishmaniasis, Visceral/transmission ; Male ; Polymerase Chain Reaction ; Psychodidae/*microbiology ; RNA, Bacterial/*genetics ; RNA, Ribosomal, 16S/*genetics ; Sequence Analysis, RNA ; Wolbachia/*isolation & purification ; }, abstract = {A polymerase chain reaction-based method was used to screen sandflies for infection with Wolbachia (Rickettsiales: Rickettsiaceae), an intracellular bacterial endosymbiont found in many arthropods and filarial hosts. Positive results were obtained in five of 200 field-collected sandflies and were confirmed by sequencing. All sandflies were Lutzomyia longipalpis (Diptera: Psychodidae) captured in a region endemic for visceral leishmaniasis in Brazil. This is the first study to identify Wolbachia infection in this Lutzomyia species, which is the main vector of leishmaniasis in the study area. The low infection rate found in this study (2.5%), together with the lack of detection of Wolbachia in previous studies and the diversity found in the sequences analysed, suggests horizontal transmission to these sandflies.}, } @article {pmid28798323, year = {2017}, author = {Wang, L and Zhao, H and Oliva, SM and Zhu, H}, title = {Modeling the transmission and control of Zika in Brazil.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {7721}, pmid = {28798323}, issn = {2045-2322}, support = {//CIHR/Canada ; }, mesh = {Aedes/microbiology/virology ; Algorithms ; Animals ; Brazil/epidemiology ; Disease Notification ; Female ; Humans ; Male ; Markov Chains ; *Models, Theoretical ; Population Surveillance ; Seasons ; Wolbachia ; *Zika Virus ; Zika Virus Infection/epidemiology/*prevention & control/*transmission/virology ; }, abstract = {Zika virus, a reemerging mosquito-borne flavivirus, started spread across Central and Southern America and more recently to North America. The most serious impacted country is Brazil. Based on the transmission mechanism of the virus and assessment of the limited data on the reported suspected cases, we establish a dynamical model which allows us to estimate the basic reproduction number R 0 = 2.5020. The wild spreading of the virus make it a great challenge to public health to control and prevention of the virus. We formulate two control models to study the impact of releasing transgenosis mosquitoes (introducing bacterium Wolbachia into Aedes aegypti) on the transmission of Zika virus in Brazil. Our models and analysis suggest that simultaneously releasing Wolbachia-harboring female and male mosquitoes will achieve the target of population replacement, while releasing only Wolbachia-harboring male mosquitoes will suppress or even eradicate wild mosquitoes eventually. We conclude that only releasing male Wolbachia mosquitoes is a better strategy for control the spreading of Zika virus in Brazil.}, } @article {pmid28793927, year = {2017}, author = {Duan, DY and Liu, GH and Cheng, TY and Wang, YQ}, title = {Microbial population analysis of the midgut of Melophagus ovinus via high-throughput sequencing.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {382}, pmid = {28793927}, issn = {1756-3305}, mesh = {Acinetobacter/genetics/isolation & purification ; Animals ; Bartonella/genetics/isolation & purification ; DNA, Bacterial/chemistry ; Digestive System/microbiology ; Diptera/*microbiology ; Female ; High-Throughput Nucleotide Sequencing ; Male ; *Microbial Consortia/genetics ; Proteobacteria/classification/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Melophagus ovinus, one of the most common haematophagous ectoparasites of sheep, can cause anaemia and reductions in weight gain, wool growth and hide value. However, no information is available about the microfloral structure of the midgut of this ectoparasite. In the present study, we investigated the microbial community structure of the midgut contents of fully engorged female and male M. ovinus using Illumina HiSeq.

RESULTS: The phylum showing the highest abundance was Proteobacteria (99.9%). The dominant bacterial genera in females and males were Bartonella, Arsenophonus and Wolbachia. Some less abundant bacterial genera were also detected, including Enterobacter, Acinetobacter, Halomonas, Shewanella, Bacillus and Staphylococcus.

CONCLUSIONS: Bartonella, Arsenophonus and Wolbachia were the dominant bacterial genera in the midgut of female and male M. ovinus. Although detected, Enterobacter, Acinetobacter, Halomonas, Shewanella, Bacillus and Staphylococcus showed low abundances. Importantly, this is the first report of the presence of Arsenophonus, Wolbachia, Enterobacter, Halomonas, Shewanella, Bacillus and Staphylococcus in the midgut of M. ovinus.}, } @article {pmid28789848, year = {2017}, author = {Kageyama, D and Wang, CH and Hatakeyama, M}, title = {Wolbachia infections of the butterfly Eurema mandarina interfere with embryonic development of the sawfly Athalia rosae.}, journal = {Journal of invertebrate pathology}, volume = {150}, number = {}, pages = {76-81}, doi = {10.1016/j.jip.2017.08.003}, pmid = {28789848}, issn = {1096-0805}, mesh = {Animals ; Bacterial Infections/*veterinary ; Butterflies/*microbiology ; Embryonic Development/*physiology ; Female ; Hymenoptera/*microbiology ; Male ; Wolbachia ; }, abstract = {Although maternally transmitted microorganisms such as Wolbachia are well known to have a variety of effects on the reproduction of diverse insect species, little is known about the underlying mechanisms of actions. Artificial transfer of Wolbachia between taxonomically distant host species may provide insights into Wolbachia-induced manipulations of hosts. Here we performed a cross-order transfer of feminizing Wolbachia derived from a butterfly, Eurema mandarina. The Wolbachia were propagated in the Eurema hecabe cell line, called NTU-YB, and then used to inject prepupal/pupal females of a Wolbachia-free hymenopteran sawfly, Athalia rosae. The 14 females that emerged as adults looked morphologically and behaviorally healthy, and ovarian development appeared normal on dissection. However, in contrast to the control, none of the 333 eggs harbored by the seven Wolbachia-injected females developed successfully. Similarly, none of the 140 eggs laid on host plant by the four Wolbachia-injected females, which were mated with males, showed any signs of development. Wolbachia infection was detected from whole-body samples of the inoculated individuals, but not from the eggs they produced. Disruption of embryonic development despite the absence of Wolbachia in the egg cytoplasm may represent a new phenotype involving maternal effects that result in female sterility.}, } @article {pmid28786370, year = {2018}, author = {Zélé, F and Denoyelle, J and Duron, O and Rivero, A}, title = {Can Wolbachia modulate the fecundity costs of Plasmodium in mosquitoes?.}, journal = {Parasitology}, volume = {145}, number = {6}, pages = {775-782}, doi = {10.1017/S0031182017001330}, pmid = {28786370}, issn = {1469-8161}, mesh = {Animals ; Coinfection/microbiology/parasitology ; Culex/parasitology ; Culicidae/*microbiology/physiology ; Fertility ; *Host-Parasite Interactions ; Malaria/transmission ; Mosquito Vectors/*microbiology/physiology ; Plasmodium/pathogenicity ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Vertically transmitted parasites (VTPs) such as Wolbachia are expected not only to minimize the damage they inflict on their hosts, but also to protect their hosts against the damaging effects of coinfecting parasites. By modifying the fitness costs of the infection, VTPs can therefore play an important role in the evolution and epidemiology of infectious diseases.Using a natural system, we explore the effects of a Wolbachia-Plasmodium co-infection on mosquito fecundity. While Plasmodium is known to frequently express its virulence by partially castrating its mosquito vectors, the effects of Wolbachia infections on mosquito fecundity are, in contrast, highly variable. Here, we show that Plasmodium drastically decreases the fecundity of mosquitoes by ca. 20%, and we provide the first evidence that this decrease is independent of the parasite's burden. Wolbachia, on the other hand, increases fecundity by roughly 10%, but does not alter the tolerance (fecundity-burden relationship) of mosquitoes to Plasmodium infection.Although Wolbachia-infected mosquitoes fare overall better than uninfected ones, Wolbachia does not confer a sufficiently high reproductive boost to mosquitoes to compensate for the reproductive losses inflicted by Plasmodium. We discuss the potential mechanisms and implications underlying the conflicting effects of these two parasites on mosquito reproduction.}, } @article {pmid28776863, year = {2017}, author = {Ikeda, T and Uchiyama, I and Iwasaki, M and Sasaki, T and Nakagawa, M and Okita, K and Masui, S}, title = {Artificial acceleration of mammalian cell reprogramming by bacterial proteins.}, journal = {Genes to cells : devoted to molecular & cellular mechanisms}, volume = {22}, number = {10}, pages = {918-928}, doi = {10.1111/gtc.12519}, pmid = {28776863}, issn = {1365-2443}, mesh = {Animals ; Cell Line ; Cellular Reprogramming/*genetics ; Cytoskeleton/metabolism ; Fibroblasts/cytology/metabolism ; *Genes, Bacterial ; Kruppel-Like Factor 4 ; Kruppel-Like Transcription Factors/genetics/metabolism ; Mice ; Neural Stem Cells/*cytology/metabolism ; Octamer Transcription Factor-3/genetics/metabolism ; Pluripotent Stem Cells/*cytology/metabolism ; Proto-Oncogene Proteins c-myc/genetics/metabolism ; SOXB1 Transcription Factors/genetics/metabolism ; Wolbachia/genetics ; }, abstract = {The molecular mechanisms of cell reprogramming and differentiation involve various signaling factors. Small molecule compounds have been identified to artificially influence these factors through interacting cellular proteins. Although such small molecule compounds are useful to enhance reprogramming and differentiation and to show the mechanisms that underlie these events, the screening usually requires a large number of compounds to identify only a very small number of hits (e.g., one hit among several tens of thousands of compounds). Here, we show a proof of concept that xenospecific gene products can affect the efficiency of cell reprogramming to pluripotency. Thirty genes specific for the bacterium Wolbachia pipientis were forcibly expressed individually along with reprogramming factors (Oct4, Sox2, Klf4 and c-Myc) that can generate induced pluripotent stem cells in mammalian cells, and eight were found to affect the reprogramming efficiency either positively or negatively (hit rate 26.7%). Mechanistic analysis suggested one of these proteins interacted with cytoskeleton to promote reprogramming. Our results raise the possibility that xenospecific gene products provide an alternative way to study the regulatory mechanism of cell identity.}, } @article {pmid28770863, year = {2017}, author = {Marris, E}, title = {Bacteria could be key to freeing South Pacific of mosquitoes.}, journal = {Nature}, volume = {548}, number = {7665}, pages = {17-18}, doi = {10.1038/548017a}, pmid = {28770863}, issn = {1476-4687}, mesh = {Aedes/*microbiology ; Animals ; Female ; Larva/microbiology ; Male ; Mosquito Vectors/*microbiology ; Pacific Islands ; Survival Rate ; Time Factors ; Wolbachia/classification/*pathogenicity ; }, } @article {pmid28753988, year = {2017}, author = {Cordaux, R and Gilbert, C}, title = {Evolutionary Significance of Wolbachia-to-Animal Horizontal Gene Transfer: Female Sex Determination and the f Element in the Isopod Armadillidium vulgare.}, journal = {Genes}, volume = {8}, number = {7}, pages = {}, pmid = {28753988}, issn = {2073-4425}, abstract = {An increasing number of horizontal gene transfer (HGT) events from bacteria to animals have been reported in the past years, many of which involve Wolbachia bacterial endosymbionts and their invertebrate hosts. Most transferred Wolbachia genes are neutrally-evolving fossils embedded in host genomes. A remarkable case of Wolbachia HGT for which a clear evolutionary significance has been demonstrated is the "f element", a nuclear Wolbachia insert involved in female sex determination in the terrestrial isopod Armadillidium vulgare. The f element represents an instance of bacteria-to-animal HGT that has occurred so recently that it was possible to infer the donor (feminizing Wolbachia closely related to the wVulC Wolbachia strain of A. vulgare) and the mechanism of integration (a nearly complete genome inserted by micro-homology-mediated recombination). In this review, we summarize our current knowledge of the f element and discuss arising perspectives regarding female sex determination, unstable inheritance, population dynamics and the molecular evolution of the f element. Overall, the f element unifies three major areas in evolutionary biology: symbiosis, HGT and sex determination. Its characterization highlights the tremendous impact sex ratio distorters can have on the evolution of sex determination mechanisms and sex chromosomes in animals and plants.}, } @article {pmid28747221, year = {2017}, author = {Montoya-Alonso, JA and Morchón, R and Falcón-Cordón, Y and Falcón-Cordón, S and Simón, F and Carretón, E}, title = {Prevalence of heartworm in dogs and cats of Madrid, Spain.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {354}, pmid = {28747221}, issn = {1756-3305}, mesh = {Animals ; Antigens, Helminth/blood ; Cat Diseases/*epidemiology/parasitology ; Cats/parasitology ; Cities ; Dirofilaria immitis/isolation & purification ; Dirofilariasis/diagnosis/*epidemiology/parasitology ; Dog Diseases/*epidemiology/parasitology ; Dogs/parasitology ; Female ; Male ; Prevalence ; Seroepidemiologic Studies ; Spain/epidemiology ; }, abstract = {BACKGROUND: Dirofilaria immitis causes heartworm disease, a chronic and potentially fatal cardiopulmonary disease which mainly affects dogs and cats. It is present in most of Spain, due to favourable climatic factors. Madrid, located in the centre of the Iberian Peninsula, is the most highly populated city in the country. There is a lack of current data on canine heartworm and there are no published epidemiological data regarding feline heartworm in this region, therefore the aim of this study was to assess the prevalence and current distribution of canine and feline dirofilariosis in the province of Madrid.

METHODS: Serum samples from 1716 dogs and 531 cats, from animals living in the metropolitan area of Madrid and adjacent areas, were studied. All the samples, either from cats and dogs, were tested for circulating D. immitis antigens using a commercial immunochromatographic test kit. Furthermore, to establish the seroprevalence of heartworm infection in cats, serological techniques for anti-D. immitis and anti-Wolbachia antibody detection were used.

RESULTS: Prevalence of D. immitis in the canine population of Madrid was 3%, showing an increase in comparison to previous data. The presence of heartworm in the city centre could be influenced by the presence of Urban Heat Islands, while the positive dogs from metropolitan and adjacent areas were mainly located under the influence of rivers. Regarding cats, 0.2% were positive to the antigens test and 7.3% were seropositive to both anti-D. immitis and Wolbachia surface protein antibodies, which demonstrate the presence of feline heartworm in Madrid. Seropositive cats were present in the same areas where positive dogs were found. Indoor/outdoor cats showed the highest seroprevalence whereas the lowest corresponded to indoor cats, demonstrating that prophylactic treatments should be carried out regardless of lifestyle. Infection was found in 2.2% of dogs and 6.7% of the cats < 1 year-old, which indicates that early preventive campaigns in puppies and kittens should be implemented.

CONCLUSIONS: The results point to the need for adequate prophylactic measures through the administration of macrocyclic lactones in animals living in Madrid. Veterinarians should be aware of the presence of this disease and include heartworm in the differential diagnosis when a pet presents with symptoms compatible with D. immitis.}, } @article {pmid28734268, year = {2017}, author = {Grzywacz, A and Wyborska, D and Piwczyński, M}, title = {DNA barcoding allows identification of European Fanniidae (Diptera) of forensic interest.}, journal = {Forensic science international}, volume = {278}, number = {}, pages = {106-114}, doi = {10.1016/j.forsciint.2017.06.023}, pmid = {28734268}, issn = {1872-6283}, mesh = {Animals ; *DNA Barcoding, Taxonomic ; DNA, Mitochondrial/genetics ; Databases, Genetic ; Diptera/*genetics ; Electron Transport Complex IV/*genetics ; Forensic Sciences ; Polymerase Chain Reaction ; }, abstract = {In forensic entomology practice, species identification is a prerequisite for any further analysis of collected material. Although morphology-based taxonomy may be hindered by a range of factors, these are not obstacles for a molecular identification approach, so-called DNA barcoding. The Fanniidae are a dipteran family that is attracted to and breeds in decomposing animal carrion and dead human bodies. However, morphological identification of fanniids, both at adult and immature stages, is considered to be difficult, particularly for non-experts. We investigated the usefulness of molecular taxonomy methods as an alternative/supplement for morphology-based identification in European Fanniidae of forensic interest. The material used in this study was collected from various regions in Asia, Europe and North America. We sequenced a barcode region of the mitochondrial cytochrome c oxidase subunit I (COI) in 27 species. For 13 species, including some taxa breeding in dead bodies, this study describes COI sequences for the first time. Our analysis revealed that both mini-barcode and full-length COI barcode sequences give very high specimen identification success. Despite the large number of COI barcode sequences referring to Fanniidae in the BOLD and GenBank databases, previous identification of forensically relevant Fanniidae was hindered by uneven taxonomic sampling. The majority of available sequences refer to species that are not of medico-legal interest, and many species of forensic interest are unrepresented or represented only by a single sequence. Because of erroneous data that are present in depository databases, DNA barcoding must be used with caution and cannot be considered to be the sole alternative to other identification methods. Wolbachia infections in the examined material did not disrupt specimen identification. The obtained results will facilitate precise identification of European Fanniidae of forensic interest, badly preserved material with degraded DNA, as well as matching of unidentified females and immature stages to already described specimens.}, } @article {pmid28732048, year = {2017}, author = {Silva, JBL and Magalhães Alves, D and Bottino-Rojas, V and Pereira, TN and Sorgine, MHF and Caragata, EP and Moreira, LA}, title = {Wolbachia and dengue virus infection in the mosquito Aedes fluviatilis (Diptera: Culicidae).}, journal = {PloS one}, volume = {12}, number = {7}, pages = {e0181678}, pmid = {28732048}, issn = {1932-6203}, mesh = {Aedes/*microbiology/*virology ; Animals ; Brazil ; Culicidae/*microbiology/*virology ; Dengue/virology ; Dengue Virus/*pathogenicity ; Gram-Negative Bacterial Infections/microbiology ; Insect Vectors/microbiology/virology ; Pest Control, Biological/methods ; Saliva/microbiology/virology ; Symbiosis/physiology ; Viral Load/physiology ; Virus Replication/physiology ; Wolbachia/*pathogenicity ; }, abstract = {Dengue represents a serious threat to human health, with billions of people living at risk of the disease. Wolbachia pipientis is a bacterial endosymbiont common to many insect species. Wolbachia transinfections in mosquito disease vectors have great value for disease control given the bacterium's ability to spread into wild mosquito populations, and to interfere with infections of pathogens, such as dengue virus. Aedes fluviatilis is a mosquito with a widespread distribution in Latin America, but its status as a dengue vector has not been clarified. Ae. fluviatilis is also naturally infected by the wFlu Wolbachia strain, which has been demonstrated to enhance infection with the avian malarial parasite Plasmodium gallinaceum. We performed experimental infections of Ae. fluviatilis with DENV-2 and DENV-3 isolates from Brazil via injection or oral feeding to provide insight into its competence for the virus. We also examined the effect of the native Wolbachia infection on the virus using a mosquito line where the wFlu infection had been cleared by antibiotic treatment. Through RT-qPCR, we observed that Ae. fluviatilis could become infected with both viruses via either method of infection, although at a lower rate than Aedes aegypti, the primary dengue vector. We then detected DENV-2 and DENV-3 in the saliva of injected mosquitoes, and observed that injection of DENV-3-infected saliva produced subsequent infections in naïve Ae. aegypti. However, across our data we observed no difference in prevalence of infection and viral load between Wolbachia-infected and -uninfected mosquitoes, suggesting that there is no effect of wFlu on dengue virus. Our results highlight that Ae. fluviatilis could potentially serve as a dengue vector under the right circumstances, although further testing is required to determine if this occurs in the field.}, } @article {pmid28724736, year = {2017}, author = {Le Clec'h, W and Dittmer, J and Raimond, M and Bouchon, D and Sicard, M}, title = {Phenotypic shift in Wolbachia virulence towards its native host across serial horizontal passages.}, journal = {Proceedings. Biological sciences}, volume = {284}, number = {1859}, pages = {}, pmid = {28724736}, issn = {1471-2954}, mesh = {Animals ; Disease Transmission, Infectious ; Isopoda/*microbiology ; Phenotype ; *Symbiosis ; Virulence ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Vertical transmission mode is predicted to decrease the virulence of symbionts. However, Wolbachia, a widespread vertically transmitted endosymbiont, exhibits both negative and beneficial effects on arthropod fitness. This 'Jekyll and Hyde' behaviour, as well as its ability to live transiently outside host cells and to establish new infections via horizontal transmission, may reflect the capacity of Wolbachia to exhibit various phenotypes depending on the prevailing environmental constraints. To study the ability of Wolbachia to readily cope with new constraints, we forced this endosymbiont to spread only via horizontal transmission. To achieve this, we performed serial horizontal transfers of haemolymph from Wolbachia-infected to naive individuals of the isopod Armadillidium vulgare. Across passages, we observed phenotypic changes in the symbiotic relationship: (i) The Wolbachia titre increased in both haemolymph and nerve cord but remained stable in ovaries; (ii) Wolbachia infection was benign at the beginning of the experiment, but highly virulent, killing most hosts after only a few passages. Such a phenotypic shift after recurrent horizontal passages demonstrates that Wolbachia can rapidly change its virulence when facing new environmental constraints. We thoroughly discuss the potential mechanism(s) underlying this phenotypic change, which are likely to be crucial for the ongoing radiation of Wolbachia in arthropods.}, } @article {pmid28702705, year = {2018}, author = {Duplouy, A and Brattström, O}, title = {Wolbachia in the Genus Bicyclus: a Forgotten Player.}, journal = {Microbial ecology}, volume = {75}, number = {1}, pages = {255-263}, pmid = {28702705}, issn = {1432-184X}, support = {266021//Suomen Akatemia/ ; }, mesh = {Animals ; Biological Evolution ; Butterflies/growth & development/*microbiology/physiology ; Phylogeny ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Bicyclus butterflies are key species for studies of wing pattern development, phenotypic plasticity, speciation and the genetics of Lepidoptera. One of the key endosymbionts in butterflies, the alpha-Proteobacterium Wolbachia pipientis, is affecting many of these biological processes; however, Bicyclus butterflies have not been investigated systematically as hosts to Wolbachia. In this study, we screen for Wolbachia infection in several Bicyclus species from natural populations across Africa as well as two laboratory populations. Out of the 24 species tested, 19 were found to be infected, and no double infection was found, but both A- and B-supergroup strains colonise this butterfly group. We also show that many of the Wolbachia strains identified in Bicyclus butterflies belong to the ST19 clonal complex. We discuss the importance of our results in regard to routinely screening for Wolbachia when using Bicyclus butterflies as the study organism of research in eco-evolutionary biology.}, } @article {pmid28690147, year = {2017}, author = {Soares, HS and Marcili, A and Barbieri, ARM and Minervino, AHH and Malheiros, AF and Gennari, SM and Labruna, MB}, title = {Novel Anaplasma and Ehrlichia organisms infecting the wildlife of two regions of the Brazilian Amazon.}, journal = {Acta tropica}, volume = {174}, number = {}, pages = {82-87}, doi = {10.1016/j.actatropica.2017.07.006}, pmid = {28690147}, issn = {1873-6254}, mesh = {Anaplasma/*genetics/*isolation & purification ; Animals ; Animals, Wild/*microbiology ; Brazil ; Ehrlichia/*genetics/*isolation & purification ; Genetic Variation ; Geography ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; }, abstract = {During 2009-2012, wild animals were sampled in the Amazon biome of Brazil. Animal tissues and blood were tested by polymerase chain reaction (PCR) assays targeting DNA of the bacterial family Anaplasmataceae (genera Anaplasma, Ehrlichia, Wolbachia) and the genus Borrelia. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. All birds and reptiles were negative by all PCR assays, as well as all mammals for the Borrelia PCR assay. Anaplasmataceae agents were searched by PCR assays targeting two different genes, the ribosomal 16S rRNA gene and the protein-coding dsb gene. Three dsb closely related haplotypes were generated from 3 white-lipped peccaries (Tayassu pecari). In a phylogenetic analysis inferred from dsb partial sequences, these haplotypes grouped with previously reported Ehrlichia haplotypes from jaguar (Panthera onca) and horse from Brazil, suggesting that they could all represent a single species, yet to be properly characterized. A unique dsb haplotype was generated from a sloth (Bradypus tridactylus), and could also represent a different Ehrlichia species. All these dsb haplotypes formed a clade sister to the Ehrlichia ruminantium clade. Three distinct 16S rRNA gene haplotypes were generated from a wild guinea pig (Cavia sp.), a woolly mouse opossum (Micoureus demerarae), and two from robust capuchin monkeys (Sapajus sp.). In a phylogenetic analysis inferred from 16S rRNA gene partial sequence, these haplotypes grouped within the Wolbachia clade, and are likely to represent Wolbachia organisms that were infecting invertebrate metazoarians (e.g., filarids) associated with the sampled mammals. Two deer (Mazama americana) samples yielded two distinct 16S rRNA gene sequences, one identical to several sequences of Anaplasma bovis, and an unique sequence that grouped in a clade with different Anaplasma species. Our results indicate that a variety of genetically distinct Anaplasmataceae organisms, including potentially new Ehrlichia species, circulate under natural conditions in the Amazonian wildlife.}, } @article {pmid28680117, year = {2017}, author = {Doudoumis, V and Blow, F and Saridaki, A and Augustinos, A and Dyer, NA and Goodhead, I and Solano, P and Rayaisse, JB and Takac, P and Mekonnen, S and Parker, AG and Abd-Alla, AMM and Darby, A and Bourtzis, K and Tsiamis, G}, title = {Challenging the Wigglesworthia, Sodalis, Wolbachia symbiosis dogma in tsetse flies: Spiroplasma is present in both laboratory and natural populations.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {4699}, pmid = {28680117}, issn = {2045-2322}, support = {BB/J017698/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/K501773/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Animals, Wild/microbiology/parasitology ; Enterobacteriaceae/classification/genetics/*isolation & purification/physiology ; Female ; High-Throughput Nucleotide Sequencing ; Male ; Multilocus Sequence Typing ; Ovary/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, RNA ; Species Specificity ; Spiroplasma/classification/genetics/*isolation & purification/physiology ; Symbiosis ; Testis/microbiology ; Tissue Distribution ; Tsetse Flies/classification/growth & development/*microbiology/*parasitology ; Wigglesworthia/classification/genetics/*isolation & purification/physiology ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Profiling of wild and laboratory tsetse populations using 16S rRNA gene amplicon sequencing allowed us to examine whether the "Wigglesworthia-Sodalis-Wolbachia dogma" operates across species and populations. The most abundant taxa, in wild and laboratory populations, were Wigglesworthia (the primary endosymbiont), Sodalis and Wolbachia as previously characterized. The species richness of the microbiota was greater in wild than laboratory populations. Spiroplasma was identified as a new symbiont exclusively in Glossina fuscipes fuscipes and G. tachinoides, members of the palpalis sub-group, and the infection prevalence in several laboratory and natural populations was surveyed. Multi locus sequencing typing (MLST) analysis identified two strains of tsetse-associated Spiroplasma, present in G. f. fuscipes and G. tachinoides. Spiroplasma density in G. f. fuscipes larva guts was significantly higher than in guts from teneral and 15-day old male and female adults. In gonads of teneral and 15-day old insects, Spiroplasma density was higher in testes than ovaries, and was significantly higher density in live versus prematurely deceased females indicating a potentially mutualistic association. Higher Spiroplasma density in testes than in ovaries was also detected by fluorescent in situ hybridization in G. f. fuscipes.}, } @article {pmid28667411, year = {2018}, author = {Szklarzewicz, T and Kalandyk-Kołodziejczyk, M and Michalik, K and Jankowska, W and Michalik, A}, title = {Symbiotic microorganisms in Puto superbus (Leonardi, 1907) (Insecta, Hemiptera, Coccomorpha: Putoidae).}, journal = {Protoplasma}, volume = {255}, number = {1}, pages = {129-138}, pmid = {28667411}, issn = {1615-6102}, support = {IP2015050374//Ministerstwo Nauki i Szkolnictwa Wyższego/ ; }, mesh = {Animals ; Hemiptera/*chemistry ; Phylogeny ; Symbiosis ; }, abstract = {The scale insect Puto superbus (Putoidae) lives in mutualistic symbiotic association with bacteria. Molecular phylogenetic analyses have revealed that symbionts of P. superbus belong to the gammaproteobacterial genus Sodalis. In the adult females, symbionts occur both in the bacteriocytes constituting compact bacteriomes and in individual bacteriocytes, which are dispersed among ovarioles. The bacteriocytes also house a few small, rod-shaped Wolbachia bacteria in addition to the numerous large, elongated Sodalis-allied bacteria. The symbiotic microorganisms are transovarially transmitted from generation to generation. In adult females which have choriogenic oocytes in the ovarioles, the bacteriocytes gather around the basal part of the tropharium. Next, the entire bacteriocytes pass through the follicular epithelium surrounding the neck region of the ovariole and enter the space between oocyte and follicular epithelium (perivitelline space). In the perivitelline space, the bacteriocytes assemble extracellularly in the deep depression of the oolemma at the anterior pole of the oocyte, forming a "symbiont ball".}, } @article {pmid28667019, year = {2017}, author = {Leung, W and Shaffer, CD and Chen, EJ and Quisenberry, TJ and Ko, K and Braverman, JM and Giarla, TC and Mortimer, NT and Reed, LK and Smith, ST and Robic, S and McCartha, SR and Perry, DR and Prescod, LM and Sheppard, ZA and Saville, KJ and McClish, A and Morlock, EA and Sochor, VR and Stanton, B and Veysey-White, IC and Revie, D and Jimenez, LA and Palomino, JJ and Patao, MD and Patao, SM and Himelblau, ET and Campbell, JD and Hertz, AL and McEvilly, MF and Wagner, AR and Youngblom, J and Bedi, B and Bettincourt, J and Duso, E and Her, M and Hilton, W and House, S and Karimi, M and Kumimoto, K and Lee, R and Lopez, D and Odisho, G and Prasad, R and Robbins, HL and Sandhu, T and Selfridge, T and Tsukashima, K and Yosif, H and Kokan, NP and Britt, L and Zoellner, A and Spana, EP and Chlebina, BT and Chong, I and Friedman, H and Mammo, DA and Ng, CL and Nikam, VS and Schwartz, NU and Xu, TQ and Burg, MG and Batten, SM and Corbeill, LM and Enoch, E and Ensign, JJ and Franks, ME and Haiker, B and Ingles, JA and Kirkland, LD and Lorenz-Guertin, JM and Matthews, J and Mittig, CM and Monsma, N and Olson, KJ and Perez-Aragon, G and Ramic, A and Ramirez, JR and Scheiber, C and Schneider, PA and Schultz, DE and Simon, M and Spencer, E and Wernette, AC and Wykle, ME and Zavala-Arellano, E and McDonald, MJ and Ostby, K and Wendland, P and DiAngelo, JR and Ceasrine, AM and Cox, AH and Docherty, JEB and Gingras, RM and Grieb, SM and Pavia, MJ and Personius, CL and Polak, GL and Beach, DL and Cerritos, HL and Horansky, EA and Sharif, KA and Moran, R and Parrish, S and Bickford, K and Bland, J and Broussard, J and Campbell, K and Deibel, KE and Forka, R and Lemke, MC and Nelson, MB and O'Keeffe, C and Ramey, SM and Schmidt, L and Villegas, P and Jones, CJ and Christ, SL and Mamari, S and Rinaldi, AS and Stity, G and Hark, AT and Scheuerman, M and Silver Key, SC and McRae, BD and Haberman, AS and Asinof, S and Carrington, H and Drumm, K and Embry, T and McGuire, R and Miller-Foreman, D and Rosen, S and Safa, N and Schultz, D and Segal, M and Shevin, Y and Svoronos, P and Vuong, T and Skuse, G and Paetkau, DW and Bridgman, RK and Brown, CM and Carroll, AR and Gifford, FM and Gillespie, JB and Herman, SE and Holtcamp, KL and Host, MA and Hussey, G and Kramer, DM and Lawrence, JQ and Martin, MM and Niemiec, EN and O'Reilly, AP and Pahl, OA and Quintana, G and Rettie, EAS and Richardson, TL and Rodriguez, AE and Rodriguez, MO and Schiraldi, L and Smith, JJ and Sugrue, KF and Suriano, LJ and Takach, KE and Vasquez, AM and Velez, X and Villafuerte, EJ and Vives, LT and Zellmer, VR and Hauke, J and Hauser, CR and Barker, K and Cannon, L and Parsamian, P and Parsons, S and Wichman, Z and Bazinet, CW and Johnson, DE and Bangura, A and Black, JA and Chevee, V and Einsteen, SA and Hilton, SK and Kollmer, M and Nadendla, R and Stamm, J and Fafara-Thompson, AE and Gygi, AM and Ogawa, EE and Van Camp, M and Kocsisova, Z and Leatherman, JL and Modahl, CM and Rubin, MR and Apiz-Saab, SS and Arias-Mejias, SM and Carrion-Ortiz, CF and Claudio-Vazquez, PN and Espada-Green, DM and Feliciano-Camacho, M and Gonzalez-Bonilla, KM and Taboas-Arroyo, M and Vargas-Franco, D and Montañez-Gonzalez, R and Perez-Otero, J and Rivera-Burgos, M and Rivera-Rosario, FJ and Eisler, HL and Alexander, J and Begley, SK and Gabbard, D and Allen, RJ and Aung, WY and Barshop, WD and Boozalis, A and Chu, VP and Davis, JS and Duggal, RN and Franklin, R and Gavinski, K and Gebreyesus, H and Gong, HZ and Greenstein, RA and Guo, AD and Hanson, C and Homa, KE and Hsu, SC and Huang, Y and Huo, L and Jacobs, S and Jia, S and Jung, KL and Wai-Chee Kong, S and Kroll, MR and Lee, BM and Lee, PF and Levine, KM and Li, AS and Liu, C and Liu, MM and Lousararian, AP and Lowery, PB and Mallya, AP and Marcus, JE and Ng, PC and Nguyen, HP and Patel, R and Precht, H and Rastogi, S and Sarezky, JM and Schefkind, A and Schultz, MB and Shen, D and Skorupa, T and Spies, NC and Stancu, G and Vivian Tsang, HM and Turski, AL and Venkat, R and Waldman, LE and Wang, K and Wang, T and Wei, JW and Wu, DY and Xiong, DD and Yu, J and Zhou, K and McNeil, GP and Fernandez, RW and Menzies, PG and Gu, T and Buhler, J and Mardis, ER and Elgin, SCR}, title = {Retrotransposons Are the Major Contributors to the Expansion of the Drosophila ananassae Muller F Element.}, journal = {G3 (Bethesda, Md.)}, volume = {7}, number = {8}, pages = {2439-2460}, pmid = {28667019}, issn = {2160-1836}, support = {U54 HG003079/HG/NHGRI NIH HHS/United States ; 52007051/HHMI/Howard Hughes Medical Institute/United States ; }, mesh = {Animals ; Base Composition/genetics ; Base Sequence ; Chromosomes/*genetics ; Codon/genetics ; Drosophila/*genetics ; Female ; Gene Expression Profiling ; Genes, Insect ; Histones/metabolism ; Protein Processing, Post-Translational/genetics ; Retroelements/*genetics ; Wolbachia/genetics ; }, abstract = {The discordance between genome size and the complexity of eukaryotes can partly be attributed to differences in repeat density. The Muller F element (∼5.2 Mb) is the smallest chromosome in Drosophila melanogaster, but it is substantially larger (>18.7 Mb) in D. ananassae To identify the major contributors to the expansion of the F element and to assess their impact, we improved the genome sequence and annotated the genes in a 1.4-Mb region of the D. ananassae F element, and a 1.7-Mb region from the D element for comparison. We find that transposons (particularly LTR and LINE retrotransposons) are major contributors to this expansion (78.6%), while Wolbachia sequences integrated into the D. ananassae genome are minor contributors (0.02%). Both D. melanogaster and D. ananassae F-element genes exhibit distinct characteristics compared to D-element genes (e.g., larger coding spans, larger introns, more coding exons, and lower codon bias), but these differences are exaggerated in D. ananassae Compared to D. melanogaster, the codon bias observed in D. ananassae F-element genes can primarily be attributed to mutational biases instead of selection. The 5' ends of F-element genes in both species are enriched in dimethylation of lysine 4 on histone 3 (H3K4me2), while the coding spans are enriched in H3K9me2. Despite differences in repeat density and gene characteristics, D. ananassae F-element genes show a similar range of expression levels compared to genes in euchromatic domains. This study improves our understanding of how transposons can affect genome size and how genes can function within highly repetitive domains.}, } @article {pmid28663939, year = {2017}, author = {Lindsey, ARI and Stouthamer, R}, title = {Penetrance of symbiont-mediated parthenogenesis is driven by reproductive rate in a parasitoid wasp.}, journal = {PeerJ}, volume = {5}, number = {}, pages = {e3505}, pmid = {28663939}, issn = {2167-8359}, abstract = {Trichogramma wasps are tiny parasitoids of lepidopteran eggs, used extensively for biological control. They are often infected with the bacterial symbiont Wolbachia, which converts Trichogramma to an asexual mode of reproduction, whereby females develop from unfertilized eggs. However, this Wolbachia-induced parthenogenesis is not always complete, and previous studies have noted that infected females will produce occasional males in the lab. The conditions that reduce penetrance of the parthenogenesis phenotype are not well understood. We hypothesized that more ecologically relevant conditions of limited host access will sustain female-biased sex ratios. After restricting access to host eggs, we found a strong relationship between reproductive rate and sex ratio. By limiting reproduction to one hour a day, wasps could sustain up to 100% effective parthenogenesis for one week, with no significant impact on total fecundity. Reproductive output in the first 24-hours appears to be critical to the total sex ratio of the entire brood. Limiting oviposition in that period resulted in more effective parthenogenesis after one week, again without any significant impact on total fecundity. Our data suggest that this phenomenon may be due to the depletion of Wolbachia when oviposition occurs continuously, whereas Wolbachia titers may recover when offspring production is limited. In addition to the potential to improve mass rearing of Trichogramma for biological control, findings from this study help elucidate the context-dependent nature of a pervasive symbiotic relationship.}, } @article {pmid28659179, year = {2017}, author = {Faddeeva-Vakhrusheva, A and Kraaijeveld, K and Derks, MFL and Anvar, SY and Agamennone, V and Suring, W and Kampfraath, AA and Ellers, J and Le Ngoc, G and van Gestel, CAM and Mariën, J and Smit, S and van Straalen, NM and Roelofs, D}, title = {Coping with living in the soil: the genome of the parthenogenetic springtail Folsomia candida.}, journal = {BMC genomics}, volume = {18}, number = {1}, pages = {493}, pmid = {28659179}, issn = {1471-2164}, mesh = {Animals ; Anti-Bacterial Agents/biosynthesis ; Arthropods/*genetics/metabolism/*physiology ; Gene Rearrangement ; Gene Transfer, Horizontal ; *Genomics ; Multigene Family/genetics ; Phylogeny ; *Soil ; }, abstract = {BACKGROUND: Folsomia candida is a model in soil biology, belonging to the family of Isotomidae, subclass Collembola. It reproduces parthenogenetically in the presence of Wolbachia, and exhibits remarkable physiological adaptations to stress. To better understand these features and adaptations to life in the soil, we studied its genome in the context of its parthenogenetic lifestyle.

RESULTS: We applied Pacific Bioscience sequencing and assembly to generate a reference genome for F. candida of 221.7 Mbp, comprising only 162 scaffolds. The complete genome of its endosymbiont Wolbachia, was also assembled and turned out to be the largest strain identified so far. Substantial gene family expansions and lineage-specific gene clusters were linked to stress response. A large number of genes (809) were acquired by horizontal gene transfer. A substantial fraction of these genes are involved in lignocellulose degradation. Also, the presence of genes involved in antibiotic biosynthesis was confirmed. Intra-genomic rearrangements of collinear gene clusters were observed, of which 11 were organized as palindromes. The Hox gene cluster of F. candida showed major rearrangements compared to arthropod consensus cluster, resulting in a disorganized cluster.

CONCLUSIONS: The expansion of stress response gene families suggests that stress defense was important to facilitate colonization of soils. The large number of HGT genes related to lignocellulose degradation could be beneficial to unlock carbohydrate sources in soil, especially those contained in decaying plant and fungal organic matter. Intra- as well as inter-scaffold duplications of gene clusters may be a consequence of its parthenogenetic lifestyle. This high quality genome will be instrumental for evolutionary biologists investigating deep phylogenetic lineages among arthropods and will provide the basis for a more mechanistic understanding in soil ecology and ecotoxicology.}, } @article {pmid28655666, year = {2017}, author = {Pimenta de Oliveira, S and Dantas de Oliveira, C and Viana Sant'Anna, MR and Carneiro Dutra, HL and Caragata, EP and Moreira, LA}, title = {Wolbachia infection in Aedes aegypti mosquitoes alters blood meal excretion and delays oviposition without affecting trypsin activity.}, journal = {Insect biochemistry and molecular biology}, volume = {87}, number = {}, pages = {65-74}, doi = {10.1016/j.ibmb.2017.06.010}, pmid = {28655666}, issn = {1879-0240}, mesh = {Aedes/*microbiology/physiology ; Animals ; Blood ; Female ; Food Deprivation ; Gastrointestinal Tract/metabolism/physiology ; Gene Expression Profiling ; Humans ; Oviposition/*physiology ; Trypsin/metabolism ; *Wolbachia ; }, abstract = {Blood feeding in Aedes aegypti is essential for reproduction, but also permits the mosquito to act as a vector for key human pathogens such as the Zika and dengue viruses. Wolbachia pipientis is an endosymbiotic bacterium that can manipulate the biology of Aedes aegypti mosquitoes, making them less competent hosts for many pathogens. Yet while Wolbachia affects other aspects of host physiology, it is unclear whether it influences physiological processes associated with blood meal digestion. To that end, we examined the effects of wMel Wolbachia infection in Ae. aegypti, on survival post-blood feeding, blood meal excretion, rate of oviposition, expression levels of key genes involved in oogenesis, and activity levels of trypsin blood digestion enzymes. We observed that wMel infection altered the rate and duration of blood meal excretion, delayed the onset of oviposition and was associated with a greater number of eggs being laid later. wMel-infected Ae. aegypti also had lower levels of key yolk protein precursor genes necessary for oogenesis. However, all of these effects occurred without a change in trypsin activity. These results suggest that Wolbachia infection may disrupt normal metabolic processes associated with blood feeding and reproduction in Ae. aegypti.}, } @article {pmid28653334, year = {2017}, author = {Joanne, S and Vythilingam, I and Teoh, BT and Leong, CS and Tan, KK and Wong, ML and Yugavathy, N and AbuBakar, S}, title = {Vector competence of Malaysian Aedes albopictus with and without Wolbachia to four dengue virus serotypes.}, journal = {Tropical medicine & international health : TM & IH}, volume = {22}, number = {9}, pages = {1154-1165}, doi = {10.1111/tmi.12918}, pmid = {28653334}, issn = {1365-3156}, mesh = {*Aedes/microbiology/virology ; Animals ; Bacterial Infections/complications ; Dengue/*transmission/virology ; Dengue Virus/classification/*pathogenicity ; Humans ; *Insect Vectors ; Polymerase Chain Reaction ; *Serogroup ; *Virus Replication ; *Wolbachia ; }, abstract = {OBJECTIVE: To determine the susceptibility status of Aedes albopictus with and without Wolbachia to the four dengue virus serotypes.

METHODS: Two newly colonised colonies of Ae. albopictus from the wild were used for the study. One colony was naturally infected with Wolbachia while in the other Wolbachia was removed by tetracycline treatment. Both colonies were orally infected with dengue virus-infected fresh blood meal. Dengue virus load was measured using quantitative RT-PCR at four-time intervals in the salivary glands, midguts and ovaries.

RESULTS: Wolbachia did not significantly affect Malaysian Ae. albopictus dengue infection or the dissemination rate for all four dengue virus serotypes. Malaysian Ae. albopictus had the highest replication kinetics for DENV-1 and the highest salivary gland and midgut infection rate for DENV-4.

CONCLUSION: Wolbachia, which naturally exists in Malaysian Ae. albopictus, does not significantly affect dengue virus replication. Malaysian Ae. albopictus is susceptible to dengue virus infections and capable of transmitting dengue virus, especially DENV-1 and DENV-4. Removal of Wolbachia from Malaysian Ae. albopictus would not reduce their susceptibility status.}, } @article {pmid28645313, year = {2017}, author = {Dahmani, M and Davoust, B and Tahir, D and Raoult, D and Fenollar, F and Mediannikov, O}, title = {Molecular investigation and phylogeny of Anaplasmataceae species infecting domestic animals and ticks in Corsica, France.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {302}, pmid = {28645313}, issn = {1756-3305}, mesh = {Anaplasmataceae/*classification/genetics ; Anaplasmataceae Infections/epidemiology/microbiology/transmission/*veterinary ; Animals ; Arachnid Vectors/*microbiology ; Cattle ; Cattle Diseases/epidemiology/microbiology/parasitology ; Chaperonin 60/genetics ; DNA, Bacterial/chemistry/isolation & purification ; Dogs ; Female ; France/epidemiology ; Goat Diseases/epidemiology/microbiology ; Goats ; Horses ; Ixodidae/genetics/*microbiology ; Male ; Neglected Diseases/epidemiology/microbiology/veterinary ; *Phylogeny ; Prevalence ; RNA, Ribosomal/genetics ; RNA, Ribosomal, 23S/genetics ; Rhipicephalus/genetics/microbiology ; Sheep ; Sheep Diseases/epidemiology/microbiology ; Tick Infestations/epidemiology/parasitology/*veterinary ; }, abstract = {BACKGROUNDS: Corsica is a French island situated in the Mediterranean Sea. The island provides suitable natural conditions to study disease ecology, especially tick-borne diseases and emerging diseases in animals and ticks. The family Anaplasmataceae is a member of the order Rickettsiales; it includes the genera Anaplasma, Ehrlichia, Neorickettsia and Wolbachia. Anaplasmosis and ehrlichiosis traditionally refer to diseases caused by obligate intracellular bacteria of the genera Anaplasma and Ehrlichia. The aim of this study was to identify and estimate the prevalence of Anaplasmataceae species infecting domestic animals and ticks in Corsica.

METHODS: In this study, 458 blood samples from sheep, cattle, horses, goats, dogs, and 123 ticks removed from cattle, were collected in Corsica. Quantitative real-time PCR screening and genetic characterisation of Anaplasmataceae bacteria were based on the 23S rRNA, rpoB and groEl genes.

RESULTS: Two tick species were collected in the present study: Rhipicephalus bursa (118) and Hyalomma marginatum marginatum (5). Molecular investigation showed that 32.1% (147/458) of blood samples were positive for Anaplasmataceae infection. Anaplasma ovis was identified in 42.3% (93/220) of sheep. Anaplasma marginale was amplified from 100% (12/12) of cattle and two R. bursa (2/123). Several potentially new species were also identified: Anaplasma cf. ovis, "Candidatus Anaplasma corsicanum", "Candidatus Anaplasma mediterraneum" were amplified from 17.3% (38/220) of sheep, and Anaplasma sp. marginale-like was amplified from 80% (4/5) of goats. Finally, one R. bursa tick was found to harbour the DNA of E. canis. All samples from horses and dogs were negative for Anaplasmataceae infection.

CONCLUSIONS: To our knowledge, this study is the first epidemiological survey on Anaplasmataceae species infecting animals and ticks in Corsica and contributes toward the identification of current Anaplasmataceae species circulating in Corsica.}, } @article {pmid28642878, year = {2017}, author = {Simón, F and González-Miguel, J and Diosdado, A and Gómez, PJ and Morchón, R and Kartashev, V}, title = {The Complexity of Zoonotic Filariasis Episystem and Its Consequences: A Multidisciplinary View.}, journal = {BioMed research international}, volume = {2017}, number = {}, pages = {6436130}, pmid = {28642878}, issn = {2314-6141}, mesh = {Animals ; Climate ; Culicidae/microbiology/parasitology ; Dirofilaria/microbiology/pathogenicity ; Dog Diseases/epidemiology/*microbiology/parasitology/transmission ; Dogs ; Filariasis/epidemiology/microbiology/*parasitology/transmission ; Humans ; Wolbachia/pathogenicity ; Zoonoses/epidemiology/microbiology/*parasitology/transmission ; }, abstract = {Vector-borne transmitted helminthic zoonosis affects the health and economy of both developing and developed countries. The concept of episystem includes the set of biological, environmental, and epidemiological elements of these diseases in defined geographic and temporal scales. Dirofilariasis caused by different species of the genus Dirofilaria is a disease affecting domestic and wild canines and felines and man, transmitted by different species of culicid mosquitoes. This complexity is increased because Dirofilaria species harbor intracellular symbiont Wolbachia bacteriae, which play a key role in the embryogenesis and development of dirofilariae and in the inflammatory pathology of the disease. In addition, the vector transmission makes the dirofilariasis susceptible to the influence of the climate and its variations. The present review addresses the analysis of dirofilariasis from the point of view of the episystem, analyzing the complex network of interactions established between biological components, climate, and factors related to human activity, as well as the different problems they pose. The progress of knowledge on human and animal dirofilariasis is largely due to the multidisciplinary approach. Nevertheless, different aspects of the disease need to continue being investigated and cooperation between countries and specialists involved should be intensified.}, } @article {pmid28640457, year = {2017}, author = {Ferla, MP and Brewster, JL and Hall, KR and Evans, GB and Patrick, WM}, title = {Primordial-like enzymes from bacteria with reduced genomes.}, journal = {Molecular microbiology}, volume = {105}, number = {4}, pages = {508-524}, doi = {10.1111/mmi.13737}, pmid = {28640457}, issn = {1365-2958}, mesh = {Alanine/metabolism ; Amino Acid Sequence ; Catalytic Domain ; Enzymes/*genetics ; Escherichia coli/genetics ; Genome/genetics ; Genome, Bacterial/genetics ; Lyases/*genetics/metabolism ; Metabolic Networks and Pathways ; Thermotoga maritima/genetics ; Wolbachia/genetics ; }, abstract = {The first cells probably possessed rudimentary metabolic networks, built using a handful of multifunctional enzymes. The promiscuous activities of modern enzymes are often assumed to be relics of this primordial era; however, by definition these activities are no longer physiological. There are many fewer examples of enzymes using a single active site to catalyze multiple physiologically-relevant reactions. Previously, we characterized the promiscuous alanine racemase (ALR) activity of Escherichia coli cystathionine β-lyase (CBL). Now we have discovered that several bacteria with reduced genomes lack alr, but contain metC (encoding CBL). We characterized the CBL enzymes from three of these: Pelagibacter ubique, the Wolbachia endosymbiont of Drosophila melanogaster (wMel) and Thermotoga maritima. Each is a multifunctional CBL/ALR. However, we also show that CBL activity is no longer required in these bacteria. Instead, the wMel and T. maritima enzymes are physiologically bi-functional alanine/glutamate racemases. They are not highly active, but they are clearly sufficient. Given the abundance of the microorganisms using them, we suggest that much of the planet's biochemistry is carried out by enzymes that are quite different from the highly-active exemplars usually found in textbooks. Instead, primordial-like enzymes may be an essential part of the adaptive strategy associated with streamlining.}, } @article {pmid28636152, year = {2017}, author = {Zhang, H and Guiguet, A and Dubreuil, G and Kisiala, A and Andreas, P and Emery, RJN and Huguet, E and Body, M and Giron, D}, title = {Dynamics and origin of cytokinins involved in plant manipulation by a leaf-mining insect.}, journal = {Insect science}, volume = {24}, number = {6}, pages = {1065-1078}, doi = {10.1111/1744-7917.12500}, pmid = {28636152}, issn = {1744-7917}, mesh = {Animals ; Cytokinins/*metabolism ; *Host-Parasite Interactions ; Larva/physiology ; Malus/*metabolism ; Moths/*physiology ; Plant Leaves/metabolism ; }, abstract = {Several herbivorous insects and plant-associated microorganisms control the phytohormonal balance, thus enabling them to successfully exploit the plant by inhibiting plant defenses and withdrawing plant resources for their own benefit. The leaf-mining moth Phyllonorycter blancardella modifies the cytokinin (CK) profile of mined leaf-tissues, and the insect symbiotic bacteria Wolbachia is involved in the plant manipulation to the benefit of the insect host. To gain a deeper understanding into the possible origin and dynamics of CKs, we conducted an extensive characterization of CKs in larvae and in infected apple leaves. Our results show the enhanced CK levels in mines, both on green and yellow leaves, allowing insects to control their nutritional supply under fluctuating environmental conditions. The spatial distribution of CKs within the mined leaves shows that hormone manipulation is strictly limited to the mine suggesting the absence of CK translocation from distant leaf areas toward the insect feeding site. Mass spectrometry analyses reveal that major CK types accumulating in mines and larvae are similar to what is observed for most gall-inducers, suggesting that strategies underlying the plant manipulation may be shared between herbivorous insects with distinct life histories. Results further show that CKs are detected in the highest levels in larvae, reinforcing our hypothesis that CKs accumulating in the mines originate from the insect itself. Presence of bacteria-specific methylthio-CKs is consistent with previous results suggesting that insect bacterial symbionts contribute to the observed phenotype. Our study provides key findings toward the understanding of molecular mechanisms underlying this intricate plant-insect-microbe interaction.}, } @article {pmid28632769, year = {2017}, author = {Kruse, A and Fattah-Hosseini, S and Saha, S and Johnson, R and Warwick, E and Sturgeon, K and Mueller, L and MacCoss, MJ and Shatters, RG and Cilia Heck, M}, title = {Combining 'omics and microscopy to visualize interactions between the Asian citrus psyllid vector and the Huanglongbing pathogen Candidatus Liberibacter asiaticus in the insect gut.}, journal = {PloS one}, volume = {12}, number = {6}, pages = {e0179531}, pmid = {28632769}, issn = {1932-6203}, support = {P30 DK048520/DK/NIDDK NIH HHS/United States ; }, mesh = {Animals ; Apoptosis/genetics ; Citric Acid Cycle/genetics ; Citrus/microbiology ; Digestive System/metabolism/microbiology ; Down-Regulation ; Hemiptera/genetics/*microbiology ; Insect Vectors/microbiology ; Microscopy, Confocal ; Mitochondria/metabolism ; Plant Diseases/microbiology ; Proteome/*analysis ; *Proteomics ; RNA/chemistry/isolation & purification/metabolism ; Rhizobiaceae/*physiology ; Sequence Analysis, RNA ; *Transcriptome ; Up-Regulation ; }, abstract = {Huanglongbing, or citrus greening disease, is an economically devastating bacterial disease of citrus. It is associated with infection by the gram-negative bacterium Candidatus Liberibacter asiaticus (CLas). CLas is transmitted by Diaphorina citri, the Asian citrus psyllid (ACP). For insect transmission to occur, CLas must be ingested during feeding on infected phloem sap and cross the gut barrier to gain entry into the insect vector. To investigate the effects of CLas exposure at the gut-pathogen interface, we performed RNAseq and mass spectrometry-based proteomics to analyze the transcriptome and proteome, respectively, of ACP gut tissue. CLas exposure resulted in changes in pathways involving the TCA cycle, iron metabolism, insecticide resistance and the insect's immune system. We identified 83 long non-coding RNAs that are responsive to CLas, two of which appear to be specific to the ACP. Proteomics analysis also enabled us to determine that Wolbachia, a symbiont of the ACP, undergoes proteome regulation when CLas is present. Fluorescent in situ hybridization (FISH) confirmed that Wolbachia and CLas inhabit the same ACP gut cells, but do not co-localize within those cells. Wolbachia cells are prevalent throughout the gut epithelial cell cytoplasm, and Wolbachia titer is more variable in the guts of CLas exposed insects. CLas is detected on the luminal membrane, in puncta within the gut epithelial cell cytoplasm, along actin filaments in the gut visceral muscles, and rarely, in association with gut cell nuclei. Our study provides a snapshot of how the psyllid gut copes with CLas exposure and provides information on pathways and proteins for targeted disruption of CLas-vector interactions at the gut interface.}, } @article {pmid28623959, year = {2017}, author = {Pacidônio, EC and Caragata, EP and Alves, DM and Marques, JT and Moreira, LA}, title = {The impact of Wolbachia infection on the rate of vertical transmission of dengue virus in Brazilian Aedes aegypti.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {296}, pmid = {28623959}, issn = {1756-3305}, mesh = {Aedes/*microbiology/virology ; Animals ; Dengue/microbiology/*transmission/virology ; Dengue Virus/*physiology ; Female ; Humans ; *Infectious Disease Transmission, Vertical ; Insect Vectors/microbiology/virology ; Larva ; Likelihood Functions ; Viral Load ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia pipientis is a common endosymbiotic bacterium of arthropods that strongly inhibits dengue virus (DENV) infection and transmission in the primary vector, the mosquito Aedes aegypti. For that reason, Wolbachia-infected Ae. aegypti are currently being released into the field as part of a novel strategy to reduce DENV transmission. However, there is evidence that DENV can be transmitted vertically from mother to progeny, and this may help the virus persist in nature in the absence of regular human transmission. The effect of Wolbachia infection on this process had not previously been examined.

RESULTS: We challenged Ae. aegypti with different Brazilian DENV isolates either by oral feeding or intrathoracic injection to ensure disseminated infection. We examined the effect of Wolbachia infection on the prevalence of DENV infection, and viral load in the ovaries. For orally infected mosquitoes, Wolbachia decreased the prevalence of infection by 71.29%, but there was no such effect when the virus was injected. Interestingly, regardless of the method of infection, Wolbachia infection strongly reduced DENV load in the ovaries. We then looked at the effect of Wolbachia on vertical transmission, where we observed only very low rates of vertical transmission. There was a trend towards lower rates in the presence of Wolbachia, with overall maximum likelihood estimate of infection rates of 5.04 per 1000 larvae for mosquitoes without Wolbachia, and 1.93 per 1000 larvae for Wolbachia-infected mosquitoes, after DENV injection. However, this effect was not statistically significant.

CONCLUSIONS: Our data support the idea that vertical transmission of DENV is rare in nature, even in the absence of Wolbachia. Indeed, we observed that vertical transmission rates were low even when the midgut barrier was bypassed, which might help to explain why we only observed a trend towards lower vertical transmission rates in the presence of Wolbachia. Nevertheless, the low prevalence of disseminated DENV infection and lower DENV load in the ovaries supports the hypothesis that the presence of Wolbachia in Ae. aegypti would have an effect on the vertical transmission of DENV in the field.}, } @article {pmid28618183, year = {2017}, author = {Hussain, M and Akutse, KS and Ravindran, K and Lin, Y and Bamisile, BS and Qasim, M and Dash, CK and Wang, L}, title = {Effects of different temperature regimes on survival of Diaphorina citri and its endosymbiotic bacterial communities.}, journal = {Environmental microbiology}, volume = {19}, number = {9}, pages = {3439-3449}, doi = {10.1111/1462-2920.13821}, pmid = {28618183}, issn = {1462-2920}, mesh = {Animals ; China ; Citrus/*parasitology ; Halomonadaceae/genetics/*growth & development ; Hemiptera/*microbiology ; Hot Temperature ; Real-Time Polymerase Chain Reaction ; Symbiosis/*physiology ; Wolbachia/genetics/*growth & development ; }, abstract = {The Asian citrus psyllid, Diaphorina citri, is a major pest of citrus and vector of citrus greening (huanglongbing) in Asian. In our field-collected psyllid samples, we discovered that Fuzhou (China) and Faisalabad (Pakistan), populations harbored an obligate primary endosymbiont Candidatus Carsonella (gen. nov.) with a single species, Candidatus Carsonella ruddii (sp. nov.) and a secondary endosymbiont, Wolbachia surface proteins (WSP) which are intracellular endosymbionts residing in the bacteriomes. Responses of these symbionts to different temperatures were examined and their host survival assessed. Diagnostic PCR assays showed that the endosymbionts infection rates were not significantly reduced in both D. citri populations after 24 h exposure to cold or heat treatments. Although quantitative PCR assays showed significant reduction of WSP relative densities at 40°C for 24 h, a substantial decrease occurred as the exposure duration increased beyond 3 days. Under the same temperature regimes, Ca. C. ruddii density was initially less affected during the first exposure day, but rapidly reduced at 3-5 days compared to WSP. However, the mortality of the psyllids increased rapidly as exposure time to heat treatment increased. The responses of the two symbionts to unfavorable temperature regimes highlight the complex host-symbionts interactions between D. citri and its associated endosymbionts.}, } @article {pmid28617844, year = {2017}, author = {Bhattacharya, T and Newton, ILG and Hardy, RW}, title = {Wolbachia elevates host methyltransferase expression to block an RNA virus early during infection.}, journal = {PLoS pathogens}, volume = {13}, number = {6}, pages = {e1006427}, pmid = {28617844}, issn = {1553-7374}, support = {R21 AI121849/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA (Cytosine-5-)-Methyltransferases/genetics/*metabolism ; Drosophila Proteins/genetics/*metabolism ; Drosophila melanogaster/*enzymology/microbiology/physiology/*virology ; Host-Pathogen Interactions ; Sindbis Virus/*physiology ; Symbiosis ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis is an intracellular endosymbiont known to confer host resistance against RNA viruses in insects. However, the causal mechanism underlying this antiviral defense remains poorly understood. To this end, we have established a robust arthropod model system to study the tripartite interaction involving Sindbis virus and Wolbachia strain wMel within its native host, Drosophila melanogaster. By leveraging the power of Drosophila genetics and a parallel, highly tractable D. melanogaster derived JW18 cell culture system, we determined that in addition to reducing infectious virus production, Wolbachia negatively influences Sindbis virus particle infectivity. This is further accompanied by reductions in viral transcript and protein levels. Interestingly, unchanged ratio of proteins to viral RNA copies suggest that Wolbachia likely does not influence the translational efficiency of viral transcripts. Additionally, expression analyses of candidate host genes revealed D. melanogaster methyltransferase gene Mt2 as an induced host factor in the presence of Wolbachia. Further characterization of viral resistance in Wolbachia-infected flies lacking functional Mt2 revealed partial recovery of virus titer relative to wild-type, accompanied by complete restoration of viral RNA and protein levels, suggesting that Mt2 acts at the stage of viral genome replication. Finally, knockdown of Mt2 in Wolbachia uninfected JW18 cells resulted in increased virus infectivity, thus demonstrating its previously unknown role as an antiviral factor against Sindbis virus. In conclusion, our findings provide evidence supporting the role of Wolbachia-modulated host factors towards RNA virus resistance in arthropods, alongside establishing Mt2's novel antiviral function against Sindbis virus in D. melanogaster.}, } @article {pmid28612849, year = {2017}, author = {Hansen, AK and Skidmore, IH}, title = {Psyllids, It's What's on the Inside That Counts: Community Cross Talk Facilitates Prophage Interactions.}, journal = {mSphere}, volume = {2}, number = {3}, pages = {}, pmid = {28612849}, issn = {2379-5042}, abstract = {Despite the availability of massive microbial community data sets (e.g., metagenomes), there is still a lack of knowledge on what molecular mechanisms facilitate cross talk between microbes and prophage within a community context. A study published in mSphere by Jain and colleagues (M. Jain, L. A. Fleites, and D. W. Gabriel, mSphere 2:e00171-17, 2017, https://doi.org/10.1128/mSphereDirect.00171-17) reports on an intriguing new twist of how a prophage of the bacterium "Candidatus Liberibacter asiaticus" may have its lytic cycle suppressed partly because of a protein that is expressed by a cooccurring bacterium, Wolbachia. Both of these microbes coexist along with other microbial tenants inside their sap-feeding insect host, a psyllid. Although these results are still preliminary and alternative hypotheses need to be tested, these results suggest an interesting new dimension on how regulation of microbial genomes occurs in a community context.}, } @article {pmid28611731, year = {2017}, author = {Crainey, JL and Hurst, J and Lamberton, PHL and Cheke, RA and Griffin, CE and Wilson, MD and de Araújo, CPM and Basáñez, MG and Post, RJ}, title = {The Genomic Architecture of Novel Simulium damnosum Wolbachia Prophage Sequence Elements and Implications for Onchocerciasis Epidemiology.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {852}, pmid = {28611731}, issn = {1664-302X}, support = {/WT_/Wellcome Trust/United Kingdom ; 77615/MRC_/Medical Research Council/United Kingdom ; }, abstract = {Research interest in Wolbachia is growing as new discoveries and technical advancements reveal the public health importance of both naturally occurring and artificial infections. Improved understanding of the Wolbachia bacteriophages (WOs) WOcauB2 and WOcauB3 [belonging to a sub-group of four WOs encoding serine recombinases group 1 (sr1WOs)], has enhanced the prospect of novel tools for the genetic manipulation of Wolbachia. The basic biology of sr1WOs, including host range and mode of genomic integration is, however, still poorly understood. Very few sr1WOs have been described, with two such elements putatively resulting from integrations at the same Wolbachia genome loci, about 2 kb downstream from the FtsZ cell-division gene. Here, we characterize the DNA sequence flanking the FtsZ gene of wDam, a genetically distinct line of Wolbachia isolated from the West African onchocerciasis vector Simulium squamosum E. Using Roche 454 shot-gun and Sanger sequencing, we have resolved >32 kb of WO prophage sequence into three contigs representing three distinct prophage elements. Spanning ≥36 distinct WO open reading frame gene sequences, these prophage elements correspond roughly to three different WO modules: a serine recombinase and replication module (sr1RRM), a head and base-plate module and a tail module. The sr1RRM module contains replication genes and a Holliday junction recombinase and is unique to the sr1 group WOs. In the extreme terminal of the tail module there is a SpvB protein homolog-believed to have insecticidal properties and proposed to have a role in how Wolbachia parasitize their insect hosts. We propose that these wDam prophage modules all derive from a single WO genome, which we have named here sr1WOdamA1. The best-match database sequence for all of our sr1WOdamA1-predicted gene sequences was annotated as of Wolbachia or Wolbachia phage sourced from an arthropod. Clear evidence of exchange between sr1WOdamA1 and other Wolbachia WO phage sequences was also detected. These findings provide insights into how Wolbachia could affect a medically important vector of onchocerciasis, with potential implications for future control methods, as well as supporting the hypothesis that Wolbachia phages do not follow the standard model of phage evolution.}, } @article {pmid28609446, year = {2017}, author = {Murray, RL and Herridge, EJ and Ness, RW and Bussière, LF}, title = {Are sex ratio distorting endosymbionts responsible for mating system variation among dance flies (Diptera: Empidinae)?.}, journal = {PloS one}, volume = {12}, number = {6}, pages = {e0178364}, pmid = {28609446}, issn = {1932-6203}, mesh = {Animals ; Biological Evolution ; Diptera/classification/*microbiology ; Female ; Host-Pathogen Interactions ; Linear Models ; Male ; Reproduction ; Rickettsia/*physiology ; Sex Ratio ; Species Specificity ; Spiroplasma/*physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Maternally inherited bacterial endosymbionts are common in many arthropod species. Some endosymbionts cause female-biased sex ratio distortion in their hosts that can result in profound changes to a host's mating behaviour and reproductive biology. Dance flies (Diptera: Empidinae) are well known for their unusual reproductive biology, including species with female-specific ornamentation and female-biased lek-like swarming behaviour. The cause of the repeated evolution of female ornaments in these flies remains unknown, but is probably associated with female-biased sex ratios in individual species. In this study we assessed whether dance flies harbour sex ratio distorting endosymbionts that might have driven these mating system evolutionary changes. We measured the incidence and prevalence of infection by three endosymbionts that are known to cause female-biased sex ratios in other insect hosts (Wolbachia, Rickettsia and Spiroplasma) across 20 species of dance flies. We found evidence of widespread infection by all three symbionts and variation in sex-specific prevalence across the taxa sampled. However, there was no relationship between infection prevalence and adult sex ratio measures and no evidence that female ornaments are associated with high prevalences of sex-biased symbiont infections. We conclude that the current distribution of endosymbiont infections is unlikely to explain the diversity in mating systems among dance fly species.}, } @article {pmid28608866, year = {2017}, author = {Jain, M and Fleites, LA and Gabriel, DW}, title = {A Small Wolbachia Protein Directly Represses Phage Lytic Cycle Genes in "Candidatus Liberibacter asiaticus" within Psyllids.}, journal = {mSphere}, volume = {2}, number = {3}, pages = {}, pmid = {28608866}, issn = {2379-5042}, abstract = {Huanglongbing (HLB) is a severe disease of citrus caused by an uncultured alphaproteobacterium "Candidatus Liberibacter asiaticus" and transmitted by Asian citrus psyllids (Diaphorina citri). Two prophage genomes, SC1 and SC2, integrated in "Ca. Liberibacter asiaticus" strain UF506 were described previously, and very similar prophages are found resident in the majority of "Ca. Liberibacter asiaticus" strains described worldwide. The SC1 lytic cycle is marked by upregulation of prophage late genes, including a functional holin (SC1_gp110); these late genes are activated when "Ca. Liberibacter asiaticus" is in planta, but not when infecting the psyllid host. We previously reported that the holin promoter is strongly and constitutively active in Liberibacter crescens (a cultured proxy for uncultured "Ca. Liberibacter asiaticus") but is suppressed in a dose-dependent manner by crude aqueous extracts from D. citri applied exogenously. Here we report that the suppressor activity of the crude psyllid extract was heat labile and abolished by proteinase K treatment, indicating a proteinaceous repressor and of a size smaller than 30 kDa. The repressor was affinity captured from D. citri aqueous extracts using biotinylated holin promoter DNA immobilized on magnetic beads and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS). Protein database interrogation was used to identify a small DNA-binding protein encoded by a gene carried by Wolbachia strain wDi, a resident endosymbiont of D. citri as the repressor. The in vitro-translated Wolbachia repressor protein was able to penetrate L. crescens cells, bind to "Ca. Liberibacter asiaticus" promoter DNA, and partially suppress holin promoter-driven β-glucuronidase (GUS) activity, indicating potential involvement of an additional interacting partner(s) or posttranslational modification(s) for complete suppression. Expression of the Wolbachia repressor protein appeared to be constitutive irrespective of "Ca. Liberibacter asiaticus" infection status of the insect host. IMPORTANCE Host acquisition of a new microbial species can readily perturb the dynamics of preexisting microbial associations. Molecular cross talk between microbial associates may be necessary for efficient resource allocation and enhanced survival. Classic examples involve quorum sensing (QS), which detects population densities and is both used and coopted to control expression of bacterial genes, including host adaptation factors. We report that a 56-amino-acid repressor protein made by the resident psyllid endosymbiont Wolbachia can enter cells of Liberibacter crescens, a cultured proxy for the uncultured psyllid endosymbiont "Ca. Liberibacter asiaticus" and repress "Ca. Liberibacter asiaticus" phage lytic cycle genes. Such repression in "Ca. Liberibacter asiaticus" may be critical to survival of both endosymbionts, since phage-mediated lysis would likely breach the immunogenic threshold of the psyllid, invoking a systemic and nonspecific innate immune reaction.}, } @article {pmid28606944, year = {2017}, author = {Emerson, KJ and Glaser, RL}, title = {Cytonuclear Epistasis Controls the Density of Symbiont Wolbachia pipientis in Nongonadal Tissues of Mosquito Culex quinquefasciatus.}, journal = {G3 (Bethesda, Md.)}, volume = {7}, number = {8}, pages = {2627-2635}, pmid = {28606944}, issn = {2160-1836}, mesh = {Animals ; Chromosome Mapping ; Culex/*microbiology ; *Epistasis, Genetic ; Female ; Gonads/*microbiology ; Inheritance Patterns/genetics ; Male ; Quantitative Trait Loci/genetics ; Symbiosis/*genetics ; Wolbachia/*genetics/*growth & development ; }, abstract = {Wolbachia pipientis, a bacterial symbiont infecting arthropods and nematodes, is vertically transmitted through the female germline and manipulates its host's reproduction to favor infected females. Wolbachia also infects somatic tissues where it can cause nonreproductive phenotypes in its host, including resistance to viral pathogens. Wolbachia-mediated phenotypes are strongly associated with the density of Wolbachia in host tissues. Little is known, however, about how Wolbachia density is regulated in native or heterologous hosts. Here, we measure the broad-sense heritability of Wolbachia density among families in field populations of the mosquito Culex pipiens, and show that densities in ovary and nongonadal tissues of females in the same family are not correlated, suggesting that Wolbachia density is determined by distinct mechanisms in the two tissues. Using introgression analysis between two different strains of the closely related species C. quinquefasciatus, we show that Wolbachia densities in ovary tissues are determined primarily by cytoplasmic genotype, while densities in nongonadal tissues are determined by both cytoplasmic and nuclear genotypes and their epistatic interactions. Quantitative-trait-locus mapping identified two major-effect quantitative-trait loci in the C. quinquefasciatus genome explaining a combined 23% of variance in Wolbachia density, specifically in nongonadal tissues. A better understanding of how Wolbachia density is regulated will provide insights into how Wolbachia density can vary spatiotemporally in insect populations, leading to changes in Wolbachia-mediated phenotypes such as viral pathogen resistance.}, } @article {pmid28603242, year = {2017}, author = {Torres-Román, JS and Bazalar-Palacios, J and Ruiz, EF and Avilez, JL and Rodríguez-Morales, AJ}, title = {New strategies for the eradication of Aedes aegypti: what challenges do we face in Latin America?.}, journal = {Le infezioni in medicina}, volume = {25}, number = {2}, pages = {193-194}, pmid = {28603242}, issn = {2532-8689}, mesh = {*Aedes/genetics/microbiology ; Animals ; Animals, Genetically Modified ; Arbovirus Infections/prevention & control/transmission ; Endemic Diseases ; Humans ; *Insect Vectors ; Latin America ; Mosquito Control/economics/*methods ; Pest Control, Biological/economics/*methods ; Wolbachia/physiology ; }, abstract = {Not available.}, } @article {pmid28600214, year = {2017}, author = {Globisch, D and Eubanks, LM and Shirey, RJ and Pfarr, KM and Wanji, S and Debrah, AY and Hoerauf, A and Janda, KD}, title = {Validation of onchocerciasis biomarker N-acetyltyramine-O-glucuronide (NATOG).}, journal = {Bioorganic & medicinal chemistry letters}, volume = {27}, number = {15}, pages = {3436-3440}, pmid = {28600214}, issn = {1464-3405}, mesh = {Animals ; Biomarkers/urine ; Chromatography, Liquid/methods ; Glucuronides/metabolism/*urine ; Humans ; Limit of Detection ; Mass Spectrometry/*methods ; Metabolomics/methods ; Neglected Diseases/metabolism/*urine ; Onchocerca volvulus/*isolation & purification/metabolism ; Onchocerciasis/metabolism/*urine ; Tyramine/*analogs & derivatives/metabolism/urine ; }, abstract = {The Neglected Tropical Disease onchocerciasis is a parasitic disease. Despite many control programmes by the World Health Organization (WHO), large communities in West and Central Africa are still affected. Besides logistic challenges during biannual mass drug administration, the lack of a robust, point-of-care diagnostic is limiting successful eradication of onchocerciasis. Towards the implementation of a non-invasive and point-of-care diagnostic, we have recently reported the discovery of the biomarker N-acetyltyramine-O-glucuronide (NATOG) in human urine samples using a metabolomics-mining approach. NATOG's biomarker value was enhanced during an investigation in a rodent model. Herein, we further detail the specificity of NATOG in active onchocerciasis infections as well as the co-infecting parasites Loa loa and Mansonella perstans. Our results measured by liquid chromatography coupled with mass spectrometry (LC-MS) reveal elevated NATOG values in mono- and co-infection samples only in the presence of the nematode Onchocerca volvulus. Metabolic pathway investigation of l-tyrosine/tyramine in all investigated nematodes uncovered an important link between the endosymbiotic bacterium Wolbachia and O. volvulus for the biosynthesis of NATOG. Based on these extended studies, we suggest NATOG as a biomarker for tracking active onchocerciasis infections and provide a threshold concentration value of NATOG for future diagnostic tool development.}, } @article {pmid28599659, year = {2017}, author = {Duchemin, JB and Mee, PT and Lynch, SE and Vedururu, R and Trinidad, L and Paradkar, P}, title = {Zika vector transmission risk in temperate Australia: a vector competence study.}, journal = {Virology journal}, volume = {14}, number = {1}, pages = {108}, pmid = {28599659}, issn = {1743-422X}, mesh = {Animals ; Australia ; Climate ; Disease Transmission, Infectious ; Gastrointestinal Tract/*virology ; Humans ; Mosquito Vectors/*virology ; RNA, Viral/*analysis/genetics ; Real-Time Polymerase Chain Reaction ; Risk Assessment ; Saliva/*virology ; Zika Virus/genetics/*isolation & purification ; Zika Virus Infection/transmission ; }, abstract = {BACKGROUND: Zika virus is an emerging pathogen of global importance. It has been responsible for recent outbreaks in the Americas and in the Pacific region. This study assessed five different mosquito species from the temperate climatic zone in Australia and included Aedes albopictus as a potentially invasive species.

METHODS: Mosquitoes were orally challenged by membrane feeding with Zika virus strain of Cambodia 2010 origin, belonging to the Asian clade. Virus infection and dissemination were assessed by quantitative PCR on midgut and carcass after dissection. Transmission was assessed by determination of cytopathogenic effect of saliva (CPE) on Vero cells, followed by determination of 50% tissue culture infectious dose (TCID50) for CPE positive samples. Additionally, the presence of Wolbachia endosymbiont infection was assessed by qPCR and standard PCR.

RESULTS: Culex mosquitoes were found unable to present Zika virus in saliva, as demonstrated by molecular as well as virological methods. Aedes aegypti, was used as a positive control for Zika infection and showed a high level of virus infection, dissemination and transmission. Local Aedes species, Ae. notoscriptus and, to a lesser degree, Ae. camptorhynchus were found to expel virus in their saliva and contained viral nucleic acid within the midgut. Molecular assessment identified low or no dissemination for these species, possibly due to low virus loads. Ae. albopictus from Torres Strait islands origin was shown as an efficient vector. Cx quinquefasciatus was shown to harbour Wolbachia endosymbionts at high prevalence, whilst no Wolbachia was found in Cx annulirostris. The Australian Ae. albopictus population was shown to harbour Wolbachia at high frequency.

CONCLUSIONS: The risk of local Aedes species triggering large Zika epidemics in the southern parts of Australia is low. The potentially invasive Ae. albopictus showed high prevalence of virus in the saliva and constitutes a potential threat if this mosquito species becomes established in mainland Australia. Complete risk analysis of Zika transmission in the temperate zone would require an assessment of the impact of temperature on Zika virus replication within local and invasive mosquito species.}, } @article {pmid28596296, year = {2017}, author = {Camacho, M and Oliva, M and Serbus, LR}, title = {Dietary saccharides and sweet tastants have differential effects on colonization of Drosophila oocytes by Wolbachia endosymbionts.}, journal = {Biology open}, volume = {6}, number = {7}, pages = {1074-1083}, pmid = {28596296}, issn = {2046-6390}, support = {R25 GM061347/GM/NIGMS NIH HHS/United States ; }, abstract = {Wolbachia bacteria are widespread, maternally transmitted endosymbionts of insects. Maintenance of sufficient Wolbachia titer in maternal germline cells is required for transmission efficacy. The mechanisms that regulate Wolbachia titer are not well understood; however, dietary sucrose was reported to elevate oocyte Wolbachia titer in Drosophila melanogaster whereas dietary yeast decreased oocyte titer. To further investigate how oocyte Wolbachia titer is controlled, this study analyzed the response of wMel Wolbachia to diets enriched in an array of natural sugars and other sweet tastants. Confocal imaging of D. melanogaster oocytes showed that food enriched in dietary galactose, lactose, maltose and trehalose elevated Wolbachia titer. However, oocyte Wolbachia titers were unaffected by exposure to the sweet tastants lactulose, erythritol, xylitol, aspartame and saccharin as compared to the control. Oocyte size was generally non-responsive to the nutrient-altered diets. Ovary size, however, was consistently smaller in response to all sugar- and sweetener-enriched diets. Furthermore, most dietary sugars administered in tandem with dietary yeast conferred complete rescue of oocyte titer suppression by yeast. All diets dually enriched in yeast and sugar also rescued yeast-associated ovary volume changes. This indicates oocyte colonization by Wolbachia to be a nutritionally sensitive process regulated by multiple mechanistic inputs.}, } @article {pmid28592678, year = {2017}, author = {Gupta, V and Vasanthakrishnan, RB and Siva-Jothy, J and Monteith, KM and Brown, SP and Vale, PF}, title = {The route of infection determines Wolbachia antibacterial protection in Drosophila.}, journal = {Proceedings. Biological sciences}, volume = {284}, number = {1856}, pages = {}, pmid = {28592678}, issn = {1471-2954}, mesh = {Animals ; Bacterial Infections/*microbiology ; Disease Resistance ; Drosophila Proteins/physiology ; Drosophila melanogaster/*microbiology ; Male ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Bacterial symbionts are widespread among metazoans and provide a range of beneficial functions. Wolbachia-mediated protection against viral infection has been extensively demonstrated in Drosophila. In mosquitoes that are artificially transinfected with Drosophila melanogaster Wolbachia (wMel), protection from both viral and bacterial infections has been demonstrated. However, no evidence for Wolbachia-mediated antibacterial protection has been demonstrated in Drosophila to date. Here, we show that the route of infection is key for Wolbachia-mediated antibacterial protection. Drosophila melanogaster carrying Wolbachia showed reduced mortality during enteric-but not systemic-infection with the opportunist pathogen Pseudomonas aeruginosaWolbachia-mediated protection was more pronounced in male flies and is associated with increased early expression of the antimicrobial peptide Attacin A, and also increased expression of a reactive oxygen species detoxification gene (Gst D8). These results highlight that the route of infection is important for symbiont-mediated protection from infection, that Wolbachia can protect hosts by eliciting a combination of resistance and disease tolerance mechanisms, and that these effects are sexually dimorphic. We discuss the importance of using ecologically relevant routes of infection to gain a better understanding of symbiont-mediated protection.}, } @article {pmid28587661, year = {2017}, author = {Morrow, JL and Hall, AAG and Riegler, M}, title = {Symbionts in waiting: the dynamics of incipient endosymbiont complementation and replacement in minimal bacterial communities of psyllids.}, journal = {Microbiome}, volume = {5}, number = {1}, pages = {58}, pmid = {28587661}, issn = {2049-2618}, mesh = {Alphaproteobacteria/classification/isolation & purification ; Animals ; Bacteria/*classification/isolation & purification ; Gammaproteobacteria/classification/isolation & purification ; Hemiptera/classification/*growth & development/microbiology ; High-Throughput Nucleotide Sequencing/*methods ; Host Specificity ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA/*methods ; Symbiosis ; }, abstract = {BACKGROUND: Obligate bacterial primary (P-) endosymbionts that are maternally inherited and codiverge with hosts are widespread across insect lineages with nutritionally restricted diets. Secondary (S-) endosymbionts are mostly facultative, but in some hosts, they complement P-endosymbiont function and therefore become obligate. Phylogenetic evidence exists for host switching and replacement of S-endosymbionts. The community dynamics that precede endosymbiont replacement and complementation have been little studied across host species, yet they are fundamental to the evolution of endosymbiosis.

RESULTS: We performed bacterial 16S rRNA gene amplicon sequencing of 25 psyllid species (Hemiptera, Psylloidea) across different developmental stages and ecological niches by focusing on the characterisation of the bacteria other than the universally present P-endosymbiont Carsonella (Gammaproteobacteria). Most species harboured only one dominant representative of diverse gammaproteobacterial S-endosymbionts that was consistently detected across all host individuals and populations (Arsenophonus in eight species, Sodalis or Sodalis-like bacteria in four species, unclassified Enterobacteriaceae in eight species). The identity of this dominant obligate S-endosymbiont varied across closely related host species. Unexpectedly, five psyllid species had two or three co-occurring endosymbiont species other than Carsonella within all host individuals, including a Rickettsiella-like bacterium (Gammaproteobacteria) in one psyllid species. Based on standard and quantitative PCR, all psyllids carried Carsonella, at higher titres than their dominant S-endosymbionts. Some psyllids also had Alphaproteobacteria (Lariskella, Rickettsia, Wolbachia) at varying prevalence. Incidence of other bacteria, including known plant pathogens, was low. Ecological niche of gall-forming, lerp-forming and free-living psyllid species did not impact endosymbiont communities. Two flush-feeding psyllid species had population-specific differences, and this was attributable to the higher endosymbiont diversity in native ranges and the absence of some endosymbionts in invasive ranges.

CONCLUSIONS: Our data support the hypothesis of strict vertical transmission of minimal core communities of bacteria in psyllids. We also found evidence for S-endosymbiont replacement across closely related psyllid species. Multiple dominant S-endosymbionts present in some host species, including at low titre, constitute potential examples of incipient endosymbiont complementation or replacement. Our multiple comparisons of deep-sequenced minimal insect bacterial communities exposed the dynamics involved in shaping insect endosymbiosis.}, } @article {pmid28586142, year = {2018}, author = {Rull, J and Tadeo, E and Lasa, R and Rodríguez, CL and Altuzar-Molina, A and Aluja, M}, title = {Experimental hybridization and reproductive isolation between two sympatric species of tephritid fruit flies in the Anastrepha fraterculus species group.}, journal = {Insect science}, volume = {25}, number = {6}, pages = {1045-1055}, doi = {10.1111/1744-7917.12489}, pmid = {28586142}, issn = {1744-7917}, support = {//Mexican Campaña Nacional Contra Moscas de la Fruta/ ; }, mesh = {Animals ; *Hybridization, Genetic ; *Reproductive Isolation ; *Sympatry ; Tephritidae/*genetics/*physiology ; Zygote/physiology ; }, abstract = {Among tephritid fruit flies, hybridization has been found to produce local adaptation and speciation, and in the case of pest species, induce behavioral and ecological alterations that can adversely impact efficient pest management. The fraterculus species group within Anastrepha (Diptera: Tephritidae), is a rapidly radiating aggregate, which includes cryptic species complexes, numerous sister species, and several pest species. Molecular studies have highlighted the possibility of introgression between A. fraterculus and A. obliqua. Reproductive isolation has been studied among morphotypes of the A. fraterculus species complex as a tool for species delimitation. Here we examined the existence and strength of prezygotic and postzygotic isolation between sympatric populations of two closely related species within the highly derived fraterculus group (A. fraterculus and A. obliqua), coexisting in nature. Although adults of both species showed a strong tendency for assortative mating, a small proportion of hybrid pairings in both directions were observed. We also observed asymmetric postzygotic isolation, with one hybrid cross displaying a strong reduction in fecundity and F1 egg fertility. Survival was greater for the progeny of homotypic and hybrid crosses in the maternal host. There was a marked female biased sex ratio distortion for both F1 hybrid adults. Hybridization between A. fraterculus and A. obliqua in nature may be difficult but possible; these two species display stronger reproductive isolation than all pairs of species previously examined in the A. fraterculus species complex. Asymmetric postzygotic isolation is suggestive of Wolbachia mediated cytoplasmic incompatibilities that may be exploited in area-wide pest management.}, } @article {pmid28579427, year = {2017}, author = {Telschow, A and Grziwotz, F and Crain, P and Miki, T and Mains, JW and Sugihara, G and Dobson, SL and Hsieh, CH}, title = {Infections of Wolbachia may destabilize mosquito population dynamics.}, journal = {Journal of theoretical biology}, volume = {428}, number = {}, pages = {98-105}, doi = {10.1016/j.jtbi.2017.05.016}, pmid = {28579427}, issn = {1095-8541}, mesh = {Aedes/microbiology ; Animals ; Culicidae/*microbiology ; Gram-Negative Bacterial Infections/*microbiology ; Models, Biological ; Nonlinear Dynamics ; Population Dynamics ; Time Factors ; Wolbachia/*growth & development ; }, abstract = {Recent efforts in controlling mosquito-borne diseases focus on biocontrol strategies that incapacitate pathogens inside mosquitoes by altering the mosquito's microbiome. A case in point is the introduction of Wolbachia into natural mosquito populations in order to eliminate Dengue virus. However, whether this strategy can successfully control vector-borne diseases is debated; particularly, how artificial infection affects population dynamics of hosts remains unclear. Here, we show that natural Wolbachia infections are associated with unstable mosquito population dynamics by contrasting Wolbachia-infected versus uninfected cage populations of the Asian tiger mosquito (Aedes albopictus). By analyzing weekly data of adult mosquito abundances, we found that the variability of the infected populations is significantly higher than that of the uninfected. The elevated population variability is explained by increased instability in dynamics, as quantified by system nonlinearity (i.e., state-dependence). In addition, predictability of infected populations is substantially lower. A mathematical model analysis suggests that Wolbachia may alter mosquito population dynamics by modifying larval competition of hosts. These results encourage examination for effects of artificial Wolbachia establishment on mosquito populations, because an enhancement of population variability with reduced predictability could pose challenges in management. Our findings have implications for application of microbiome alterations in biocontrol programs.}, } @article {pmid28577575, year = {2017}, author = {Minard, G and Tran Van, V and Tran, FH and Melaun, C and Klimpel, S and Koch, LK and Ly Huynh Kim, K and Huynh Thi Thuy, T and Tran Ngoc, H and Potier, P and Mavingui, P and Valiente Moro, C}, title = {Identification of sympatric cryptic species of Aedes albopictus subgroup in Vietnam: new perspectives in phylosymbiosis of insect vector.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {276}, pmid = {28577575}, issn = {1756-3305}, mesh = {Aedes/anatomy & histology/*classification/genetics/*microbiology ; Animals ; Bacteria/*classification/genetics/isolation & purification ; Biodiversity ; Culicidae ; Insect Vectors/classification/*microbiology ; Microbiota ; Mitochondria ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; Vietnam ; Wolbachia/classification/pathogenicity ; }, abstract = {BACKGROUND: The Aedes (Stegomyia) albopictus subgroup includes 11 cryptic species of which Ae. albopictus is the most widely distributed. Its global expansion associated with a documented vector competence for several emerging arboviruses raise obvious concerns in the recently colonized regions. While several studies have provided important insights regarding medical importance of Ae. albopicus, the investigations of the other sibling species are scarce. In Asia, indigenous populations within the Ae. albopictus subgroup can be found in sympatry. In the present study, we aimed to describe and compare molecular, morphological and bacterial symbionts composition among sympatric individuals from the Ae. albopictus subgroup inhabiting a Vietnamese protected area.

RESULTS: Based on morphological structure of the cibarial armarture, we identified a cryptic species in the forest park at Bù Gia Mập in the south-eastern region of Vietnam. Analysis of nuclear (ITS1-5.8S-ITS2) and mitochondrial (cox1, nad5) markers confirmed the divergence between the cryptic species and Ae. albopictus. Analysis of midgut bacterial microbiota revealed a strong similarity among the two species with a notable difference; contrary to Ae. albopictus, the cryptic species did not harbour any Wolbachia infection.

CONCLUSIONS: These results could reflect either a recent invasion of Wolbachia in Ae. albopictus or alternatively a loss of this symbiont in the cryptic species. We argue that neglected species of the Ae. albopictus subgroup are of main importance in order to estimate variation of host-symbionts interactions across evolution.}, } @article {pmid28573466, year = {2018}, author = {Zheng, B and Tang, M and Yu, J and Qiu, J}, title = {Wolbachia spreading dynamics in mosquitoes with imperfect maternal transmission.}, journal = {Journal of mathematical biology}, volume = {76}, number = {1-2}, pages = {235-263}, pmid = {28573466}, issn = {1432-1416}, support = {201409945004//China Scholarship Council/International ; 11301103//National Natural Science Foundation of China/International ; 91230104//National Natural Science Foundation of China/International ; 11471085//National Natural Science Foundation of China/International ; 11631005//National Natural Science Foundation of China/International ; 11626246//National Natural Science Foundation of China/International ; Y12013131//Department of Education of Guangdong Province/International ; IRT_16R16//Program for Changjiang Scholars and Innovative Research Team in University/International ; }, mesh = {Aedes/growth & development/microbiology/virology ; Animals ; Computational Biology ; Dengue/prevention & control/transmission/virology ; Female ; Humans ; Infectious Disease Transmission, Vertical/*prevention & control/statistics & numerical data ; Longevity ; Male ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/growth & development/*microbiology/virology ; Pest Control, Biological/*methods/statistics & numerical data ; Population Dynamics/statistics & numerical data ; Wolbachia/*physiology ; }, abstract = {Mosquitoes are primary vectors of life-threatening diseases such as dengue, malaria, and Zika. A new control method involves releasing mosquitoes carrying bacterium Wolbachia into the natural areas to infect wild mosquitoes and block disease transmission. In this work, we use differential equations to describe Wolbachia spreading dynamics, focusing on the poorly understood effect of imperfect maternal transmission. We establish two useful identities and employ them to prove that the system exhibits monomorphic, bistable, and polymorphic dynamics, and give sufficient and necessary conditions for each case. The results suggest that the largest maternal transmission leakage rate supporting Wolbachia spreading does not necessarily increase with the fitness of infected mosquitoes. The bistable dynamics is defined by the existence of two stable equilibria, whose basins of attraction are divided by the separatrix of a saddle point. By exploring the analytical property of the separatrix with some sharp estimates, we find that Wolbachia in a completely infected population could be wiped out ultimately if the initial population size is small. Surprisingly, when the infection shortens the lifespan of infected females that would impede Wolbachia spreading, such a reversion phenomenon does not occur.}, } @article {pmid28570608, year = {2017}, author = {Jiggins, FM}, title = {The spread of Wolbachia through mosquito populations.}, journal = {PLoS biology}, volume = {15}, number = {6}, pages = {e2002780}, pmid = {28570608}, issn = {1545-7885}, mesh = {Aedes/growth & development/immunology/*microbiology/virology ; Animal Distribution ; Animals ; Biological Control Agents/adverse effects/*immunology/isolation & purification ; California ; Colony Collapse ; Dengue/prevention & control/transmission ; Drosophila simulans/growth & development/immunology/microbiology ; Female ; *Global Health ; Humans ; Male ; Mosquito Vectors/immunology/*microbiology/virology ; Queensland ; Rickettsiaceae Infections/immunology/microbiology/transmission ; *Symbiosis ; Wolbachia/growth & development/*immunology/isolation & purification/pathogenicity ; Zika Virus Infection/prevention & control/transmission ; }, abstract = {In many regions of the world, mosquito-borne viruses pose a growing threat to human health. As an alternative to traditional control measures, the bacterial symbiont Wolbachia has been transferred from Drosophila into the mosquito Aedes aegypti, where it can block the transmission of dengue and Zika viruses. A recent paper has reported large-scale releases of Wolbachia-infected Ae. aegypti in the city of Cairns, Australia. Wolbachia, which is maternally transmitted, invaded and spread through the populations due to a sperm-egg incompatibility called cytoplasmic incompatibility. Over a period of 2 years, a wave of Wolbachia infection slowly spread out from 2 release sites, demonstrating that it will be possible to deploy this strategy in large urban areas. In line with theoretical predictions, Wolbachia infection at a third, smaller release site collapsed due to the immigration of Wolbachia-free mosquitoes from surrounding areas. This remarkable field experiment has both validated theoretical models of Wolbachia population dynamics and demonstrated that this is a viable strategy to modify mosquito populations.}, } @article {pmid28566542, year = {2017}, author = {Miyata, M and Konagaya, T and Yukuhiro, K and Nomura, M and Kageyama, D}, title = {Wolbachia-induced meiotic drive and feminization is associated with an independent occurrence of selective mitochondrial sweep in a butterfly.}, journal = {Biology letters}, volume = {13}, number = {5}, pages = {}, pmid = {28566542}, issn = {1744-957X}, mesh = {Animals ; Butterflies ; DNA, Mitochondrial ; Female ; Feminization ; Male ; Mitochondria ; Sex Ratio ; *Wolbachia ; }, abstract = {Maternally inherited Wolbachia endosymbionts manipulate arthropod reproduction in various ways. In the butterfly Eurema mandarina, a cytoplasmic incompatibility-inducing Wolbachia strain wCI and the associated mtDNA haplotypes are known to originate from the sister species Eurema hecabe, which offered a good case study for microbe-mediated hybrid introgression. Besides wCI, some females with the Z0 karyotype harbour a distinct Wolbachia strain wFem, which causes all-female production by meiotic drive and feminization. We report that a considerable proportion of E. mandarina females (65.7%) were infected with both wCI and wFem (CF) on Tanegashima Island. While females singly infected with wCI (C) produced offspring at a 1 : 1 sex ratio, CF females produced only females. Although Z-linked sequence polymorphism showed no signs of divergence between C and CF females, mtDNA split into two discrete clades; one consisted of C females and the other CF females, both of which formed a clade with E. hecabe but not with uninfected E. mandarina This suggests that CF matrilines also, but independently, experienced a selective sweep after hybrid introgression from E. hecabe Distinct evolutionary forces were suggested to have caused C and CF matrilines to diverge, which would be irreversible because of the particular phenotype of wFem.}, } @article {pmid28558804, year = {2017}, author = {Buhagiar, TS and Devine, GJ and Ritchie, SA}, title = {Metofluthrin: investigations into the use of a volatile spatial pyrethroid in a global spread of dengue, chikungunya and Zika viruses.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {270}, pmid = {28558804}, issn = {1756-3305}, mesh = {Aedes/*drug effects/physiology/virology ; Animals ; Chikungunya Fever/*prevention & control/transmission/virology ; Chikungunya virus/physiology ; Cyclopropanes/chemistry/*pharmacology ; Dengue/prevention & control/*transmission/virology ; Dengue Virus/physiology ; Female ; Fluorobenzenes/chemistry/*pharmacology ; Insect Repellents/*pharmacology ; Insect Vectors/*drug effects/physiology/virology ; Mosquito Control/instrumentation/*methods ; Zika Virus/physiology ; Zika Virus Infection/*prevention & control/transmission/virology ; }, abstract = {BACKGROUND: Metofluthrin reduces biting activity in Aedes aegypti through the confusion, knockdown, and subsequent kill of a mosquito. A geographical spread in dengue, chikungunya, and Zika viruses, increases intervention demands. Response to a Zika outbreak may require a different strategy than dengue, as high-risk individuals, specifically pregnant women, need to be targeted.

METHODS: In semi-field conditions within a residential property in Cairns, Queensland, the impacts of metofluthrin on biting behaviour of free-flying Wolbachia-infected Ae. aegypti were evaluated.

RESULTS: Mortality in Ae. aegypti exposed to metofluthrin over a 22 h period was 100% compared to 2.7% in an untreated room. No biting activity was observed in mosquitoes up to 5 m from the emanator after 10 min of metofluthrin exposure. Use of metofluthrin reduced biting activity up to 8 m, regardless of the host's proximity (near or far) to a dark harbourage area (HA) (P < 0.0001 and P = 0.006), respectively. In the presence or absence of the metofluthrin emanator, the host was most likely bitten when located immediately next to a HA (within 1 m) versus 8 m away from the HA (P = 0.006). The addition of a ceiling fan (0.8 m/s airflow) prevented all biting activity after 10 min of metofluthrin exposure. Previously unexposed Ae. aegypti were less likely to reach the host in a metofluthrin-treated room [Formula: see text]= 31%) compared to an untreated room ([Formula: see text]) (P < 0.0001). In a treated room, if the mosquito had not reached the host within 30 s, they never would. Upon activation, the time required for metofluthrin to infiltrate protected locations within a room causing knockdown in caged mosquitoes, required more time than exposed locations (P < 0.003); however exposed and protected locations do eventually reach equilibrium, affecting mosquitoes equally throughout the room.

CONCLUSION: Metofluthrin is effective in interrupting indoor host-seeking in Ae. aegypti. Metofluthrin's efficacy is increased by centrally locating the emanator in the room, and by using a fan to increase airflow. Newly treated rooms may require a period of 2-4 h for sufficient distribution of the metofluthrin into protected locations where mosquitoes may be resting.}, } @article {pmid28557993, year = {2017}, author = {Schmidt, TL and Barton, NH and Rašić, G and Turley, AP and Montgomery, BL and Iturbe-Ormaetxe, I and Cook, PE and Ryan, PA and Ritchie, SA and Hoffmann, AA and O'Neill, SL and Turelli, M}, title = {Local introduction and heterogeneous spatial spread of dengue-suppressing Wolbachia through an urban population of Aedes aegypti.}, journal = {PLoS biology}, volume = {15}, number = {5}, pages = {e2001894}, pmid = {28557993}, issn = {1545-7885}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/growth & development/*microbiology/physiology/virology ; Animals ; *Biological Control Agents/isolation & purification ; Colony Collapse/microbiology/virology ; Computer Heuristics ; Dengue/*prevention & control/transmission/virology ; Dengue Virus/growth & development/isolation & purification/*physiology ; Disease Vectors ; Female ; Global Health ; Health Transition ; Humans ; Infection Control ; Male ; *Models, Biological ; Parks, Recreational ; Queensland ; Spatio-Temporal Analysis ; *Urbanization ; Wolbachia/growth & development/isolation & purification/*physiology ; }, abstract = {Dengue-suppressing Wolbachia strains are promising tools for arbovirus control, particularly as they have the potential to self-spread following local introductions. To test this, we followed the frequency of the transinfected Wolbachia strain wMel through Ae. aegypti in Cairns, Australia, following releases at 3 nonisolated locations within the city in early 2013. Spatial spread was analysed graphically using interpolation and by fitting a statistical model describing the position and width of the wave. For the larger 2 of the 3 releases (covering 0.97 km2 and 0.52 km2), we observed slow but steady spatial spread, at about 100-200 m per year, roughly consistent with theoretical predictions. In contrast, the smallest release (0.11 km2) produced erratic temporal and spatial dynamics, with little evidence of spread after 2 years. This is consistent with the prediction concerning fitness-decreasing Wolbachia transinfections that a minimum release area is needed to achieve stable local establishment and spread in continuous habitats. Our graphical and likelihood analyses produced broadly consistent estimates of wave speed and wave width. Spread at all sites was spatially heterogeneous, suggesting that environmental heterogeneity will affect large-scale Wolbachia transformations of urban mosquito populations. The persistence and spread of Wolbachia in release areas meeting minimum area requirements indicates the promise of successful large-scale population transformation.}, } @article {pmid28542621, year = {2017}, author = {Tamura, M and Kageyama, D and Honda, N and Fujimoto, H and Kato, A}, title = {Enzymatic activity necessary to restore the lethality due to Escherichia coli RNase E deficiency is distributed among bacteria lacking RNase E homologues.}, journal = {PloS one}, volume = {12}, number = {5}, pages = {e0177915}, pmid = {28542621}, issn = {1932-6203}, mesh = {Animals ; Computer Simulation ; Endoribonucleases/chemistry/deficiency/genetics/*metabolism ; Escherichia coli/enzymology/*genetics ; Female ; *Genetic Engineering ; Mutation ; Phenotype ; *Sequence Homology, Amino Acid ; Symbiosis ; Wolbachia/enzymology ; }, abstract = {Escherichia coli RNase E (Eco-RNase E), encoded by rne (Eco-rne), is considered the global RNA decay initiator. Although Eco-RNase E is an essential gene product in E. coli, some bacterial species, such as Bacillus subtilis, do not possess Eco-RNase E sequence homologues. B. subtilis instead possesses RNase J1/J2 (Bsu-RNase J1/J2) and RNase Y (Bsu-RNase Y) to execute RNA decay. Here we found that E. coli lacking the Eco-rne gene (Δrne E. coli) was viable conditional on M9 minimal media by introducing Bsu-RNase J1/J2 or Bsu-RNase Y. We also cloned an extremely short Eco-RNase E homologue (Wpi-RNase E) and a canonical sized Bsu-RNase J1/J2 homologue (Wpi-RNase J) from Wolbachia pipientis, an α-proteobacterial endosymbiont of arthropods. We found that Wpi-RNase J restored the colony-forming ability (CFA) of Δrne E. coli, whereas Wpi-RNase E did not. Unexpectedly, Wpi-RNase E restored defective CFA due to lack of Eco-RNase G, a paralogue of Eco-RNase E. Our results indicate that bacterial species that lack Eco-RNase E homologues or bacterial species that possess Eco-RNase E homologues which lack Eco-RNase E-like activities have a modest Eco-RNase E-like function using RNase J and/or RNase Y. These results suggest that Eco-RNase E-like activities might distribute among a wide array of bacteria and that functions of RNases may have changed dynamically during evolutionary divergence of bacterial lineages.}, } @article {pmid28542240, year = {2017}, author = {Tan, CH and Wong, PJ and Li, MI and Yang, H and Ng, LC and O'Neill, SL}, title = {wMel limits zika and chikungunya virus infection in a Singapore Wolbachia-introgressed Ae. aegypti strain, wMel-Sg.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {5}, pages = {e0005496}, pmid = {28542240}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*virology ; Animals ; Bacterial Infections ; Chikungunya Fever/*epidemiology ; Chikungunya virus/*isolation & purification ; Female ; Gastrointestinal Tract/virology ; Humans ; Male ; Microbial Interactions ; Salivary Glands/virology ; Singapore/epidemiology ; Viral Load ; Wolbachia/*growth & development ; Zika Virus/*isolation & purification ; Zika Virus Infection/*epidemiology ; }, abstract = {BACKGROUND: Zika (ZIKV) and Chikungunya (CHIKV) viruses are emerging Aedes-borne viruses that are spreading outside their known geographic range and causing wide-scale epidemics. It has been reported that these viruses can be transmitted efficiently by Ae. aegypti. Recent studies have shown that Ae. aegypti when transinfected with certain Wolbachia strains shows a reduced replication and dissemination of dengue (DENV), Chikungunya (CHIKV), and Yellow Fever (YFV) viruses. The aim of this study was to determine whether the wMel strain of Wolbachia introgressed onto a Singapore Ae. aegypti genetic background was able to limit ZIKV and CHIKV infection in the mosquito.

Five to seven-day old mosquitoes either infected or uninfected with wMel Wolbachia were orally infected with a Ugandan strain of ZIKV and several outbreak strains of CHIKV. The midgut and salivary glands of each mosquito were sampled at days 6, 9 and 13 days post infectious blood meal to determine midgut infection and salivary glands dissemination rates, respectively. In general, all wild type Ae. aegypti were found to have high ZIKV and CHIKV infections in their midguts and salivary glands, across all sampling days, compared to Wolbachia infected counterparts. Median viral titre for all viruses in Wolbachia infected mosquitoes were significantly lower across all time points when compared to wild type mosquitoes. Most significantly, all but two and one of the wMel infected mosquitoes had no detectable ZIKV and CHIKV, respectively, in their salivary glands at 14 days post-infectious blood meal.

CONCLUSIONS: Our results showed that wMel limits both ZIKV and CHIKV infection when introgressed into a Singapore Ae. aegypti genetic background. These results also strongly suggest that female Aedes aegypti carrying Wolbachia will have a reduced capacity to transmit ZIKV and CHIKV.}, } @article {pmid28534089, year = {2017}, author = {Hubert, J and Erban, T and Kopecky, J and Sopko, B and Nesvorna, M and Lichovnikova, M and Schicht, S and Strube, C and Sparagano, O}, title = {Comparison of Microbiomes between Red Poultry Mite Populations (Dermanyssus gallinae): Predominance of Bartonella-like Bacteria.}, journal = {Microbial ecology}, volume = {74}, number = {4}, pages = {947-960}, pmid = {28534089}, issn = {1432-184X}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Bartonella/classification/genetics/isolation & purification ; Czech Republic ; DNA Barcoding, Taxonomic ; *Microbiota ; Mites/growth & development/*microbiology ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Species Specificity ; }, abstract = {Blood feeding red poultry mites (RPM) serve as vectors of pathogenic bacteria and viruses among vertebrate hosts including wild birds, poultry hens, mammals, and humans. The microbiome of RPM has not yet been studied by high-throughput sequencing. RPM eggs, larvae, and engorged adult/nymph samples obtained in four poultry houses in Czechia were used for microbiome analyses by Illumina amplicon sequencing of the 16S ribosomal RNA (rRNA) gene V4 region. A laboratory RPM population was used as positive control for transcriptome analysis by pyrosequencing with identification of sequences originating from bacteria. The samples of engorged adult/nymph stages had 100-fold more copies of 16S rRNA gene copies than the samples of eggs and larvae. The microbiome composition showed differences among the four poultry houses and among observed developmental stadia. In the adults' microbiome 10 OTUs comprised 90 to 99% of all sequences. Bartonella-like bacteria covered between 30 and 70% of sequences in RPM microbiome and 25% bacterial sequences in transcriptome. The phylogenetic analyses of 16S rRNA gene sequences revealed two distinct groups of Bartonella-like bacteria forming sister groups: (i) symbionts of ants; (ii) Bartonella genus. Cardinium, Wolbachia, and Rickettsiella sp. were found in the microbiomes of all tested stadia, while Spiroplasma eriocheiris and Wolbachia were identified in the laboratory RPM transcriptome. The microbiomes from eggs, larvae, and engorged adults/nymphs differed. Bartonella-like symbionts were found in all stadia and sampling sites. Bartonella-like bacteria was the most diversified group within the RPM microbiome. The presence of identified putative pathogenic bacteria is relevant with respect to human and animal health issues while the identification of symbiontic bacteria can lead to new control methods targeting them to destabilize the arthropod host.}, } @article {pmid28533374, year = {2017}, author = {Takano, SI and Tuda, M and Takasu, K and Furuya, N and Imamura, Y and Kim, S and Tashiro, K and Iiyama, K and Tavares, M and Amaral, AC}, title = {Unique clade of alphaproteobacterial endosymbionts induces complete cytoplasmic incompatibility in the coconut beetle.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {114}, number = {23}, pages = {6110-6115}, pmid = {28533374}, issn = {1091-6490}, mesh = {Alphaproteobacteria/genetics/metabolism/*pathogenicity ; Animals ; Arthropods/genetics ; Bacteroidetes/genetics ; Biological Control Agents ; Coleoptera/metabolism/*microbiology ; Cytoplasm/microbiology ; Extrachromosomal Inheritance ; Genetic Speciation ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Reproduction ; Reproductive Isolation ; Symbiosis/physiology ; Wolbachia/metabolism ; }, abstract = {Maternally inherited bacterial endosymbionts in arthropods manipulate host reproduction to increase the fitness of infected females. Cytoplasmic incompatibility (CI) is one such manipulation, in which uninfected females produce few or no offspring when they mate with infected males. To date, two bacterial endosymbionts, Wolbachia and Cardinium, have been reported as CI inducers. Only Wolbachia induces complete CI, which causes 100% offspring mortality in incompatible crosses. Here we report a third CI inducer that belongs to a unique clade of Alphaproteobacteria detected within the coconut beetle, Brontispa longissima This beetle comprises two cryptic species, the Asian clade and the Pacific clade, which show incompatibility in hybrid crosses. Different bacterial endosymbionts, a unique clade of Alphaproteobacteria in the Pacific clade and Wolbachia in the Asian clade, induced bidirectional CI between hosts. The former induced complete CI (100% mortality), whereas the latter induced partial CI (70% mortality). Illumina MiSeq sequencing and denaturing gradient gel electrophoresis patterns showed that the predominant bacterium detected in the Pacific clade of B. longissima was this unique clade of Alphaproteobacteria alone, indicating that this endosymbiont was responsible for the complete CI. Sex distortion did not occur in any of the tested crosses. The 1,160 bp of 16S rRNA gene sequence obtained for this endosymbiont had only 89.3% identity with that of Wolbachia, indicating that it can be recognized as a distinct species. We discuss the potential use of this bacterium as a biological control agent.}, } @article {pmid28506794, year = {2017}, author = {Suh, E and Mercer, DR and Dobson, SL}, title = {Life-shortening Wolbachia infection reduces population growth of Aedes aegypti.}, journal = {Acta tropica}, volume = {172}, number = {}, pages = {232-239}, pmid = {28506794}, issn = {1873-6254}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology ; Animals ; Host-Pathogen Interactions ; Larva/microbiology ; Population Growth ; Survival Analysis ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are being introduced into natural populations of vector mosquitoes, with the goal of reducing the transmission of human diseases such as Zika and dengue fever. The successful establishment of Wolbachia infection is largely dependent on the effects of Wolbachia infection to host fitness, but the effects of Wolbachia infection on the individual life-history traits of immature mosquitoes can vary. Here, the effects of life-shortening Wolbachia (wMelPop) on population growth of infected individuals were evaluated by measuring larval survival, developmental time and adult size of Aedes aegypti in intra- (infected or uninfected only) and inter-group (mixed with infected and uninfected) larval competition assays. At low larval density conditions, the population growth of wMelPop infected and uninfected individuals was similar. At high larval densities, wMelPop infected individuals had a significantly reduced population growth rate relative to uninfected individuals, regardless of competition type. We discuss the results in relation to the invasion of the wMelPop Wolbachia infection into naturally uninfected populations.}, } @article {pmid28505369, year = {2017}, author = {Tsugeno, Y and Koyama, H and Takamatsu, T and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Identification of an Early Male-Killing Agent in the Oriental Tea Tortrix, Homona magnanima.}, journal = {The Journal of heredity}, volume = {108}, number = {5}, pages = {553-560}, doi = {10.1093/jhered/esx049}, pmid = {28505369}, issn = {1465-7333}, mesh = {Animals ; Female ; Japan ; Male ; Moths/drug effects/*microbiology/*physiology ; RNA, Ribosomal, 16S ; *Sex Ratio ; Spiroplasma/genetics/*physiology ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/physiology ; }, abstract = {Arthropods are frequently infected with inherited symbionts, which sometimes confer fitness benefits on female hosts or manipulate host reproduction. Early male killing, in which infected males die during embryogenesis, is induced by some bacteria, such as Wolbachia and Spiroplasma. A female-biased sex ratio has been found in Homona magnanima, collected from a tea plantation in Japan. Here, we examined the male-killing trait in H. magnanima and identified the agent that induces early male killing. The sex ratio distortion (SR) strain produced only females and no males, and its egg hatch rate was significantly lower than that of the normal (N) strain. The N strain was infected with only Wolbachia, whereas the SR strain was infected with both Wolbachia and Spiroplasma. Antibiotic treatment with 0.10% tetracycline restored the 1:1 sex ratio in the SR strain. Females treated with 0.05% tetracycline were positive for Spiroplasma but not for Wolbachia and showed a female-biased sex ratio, whereas Wolbachia-positive females did not revert to male killing. When inoculated with a homogenate of the SR strain female, females infected with only Spiroplasma produced female-biased offspring. Sequence analysis of the 16S rRNA gene revealed that Spiroplasma sp. of H. magnanima belonged to the ixodetis clade. These results indicate that Spiroplasma was responsible for male killing in H. magnanima. Late male killing is induced in H. magnanima by an RNA-like virus, and therefore this is the first case in which different male-killing agents expressed at different times in the life cycle have been found within one host species.}, } @article {pmid28505238, year = {2017}, author = {Karut, K and Mete Karaca, M and Döker, I and Kazak, C}, title = {Analysis of Species, Subgroups, and Endosymbionts of Bemisia tabaci (Hemiptera: Aleyrodidae) From Southwestern Cotton Fields in Turkey.}, journal = {Environmental entomology}, volume = {46}, number = {4}, pages = {1035-1040}, doi = {10.1093/ee/nvx093}, pmid = {28505238}, issn = {1938-2936}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; *Bacterial Physiological Phenomena ; Female ; Gossypium/growth & development ; Hemiptera/*classification/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; *Symbiosis ; Turkey ; }, abstract = {Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is one of the most important insect pests worldwide including Turkey. Although there are substantial data regarding species composition of Turkish B. tabaci populations, the situation is still not clear and further investigations are needed. Therefore, in this study, species and subgroups of B. tabaci collected from cotton fields in southwestern part of Turkey (Antalya, Aydın, Denizli, and Muğla) were determined using microsatellite analysis, AluI-based mtCOI polymerase chain reaction-random length polymorphism, and sequencing. Secondary endosymbionts were also determined using diagnostic species-specific PCR. Middle East Asia Minor 1 (MEAM1), Mediterranean (MED) Q1, and MED Q2 were the species and subgroups found in this study. The MED species (85.3%) were found to be more dominant than MEAM1. Species status of B. tabaci varied depending on the location. Although all samples collected from Aydın were found to be Q1, three species and subgroups were found in Muğla. Secondary endosymbionts varied according to species and subgroups. Arsenophonus was found only from Q2, while Hamiltonella was detected in MEAM1 and Q1. In addition, high Rickettsia and low Wolbachia infections were detected in MEAM1 and Q1 populations, respectively. In conclusion, for the first time, we report the presence and symbiotic communities of Q1 from Turkey. We also found that the symbiont complement of the Q1 is more congruent with Q1 from Greece than other regions of the world, which may have some interesting implications for movement of this invasive subgroup.}, } @article {pmid28490339, year = {2017}, author = {Nugapola, NWNP and De Silva, WAPP and Karunaratne, SHPP}, title = {Distribution and phylogeny of Wolbachia strains in wild mosquito populations in Sri Lanka.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {230}, pmid = {28490339}, issn = {1756-3305}, mesh = {Aedes/microbiology ; Animals ; Anopheles/microbiology ; Bacterial Outer Membrane Proteins/genetics ; Chaperonin 60/genetics ; Culex/microbiology ; Culicidae/*microbiology ; Mosquito Vectors/*microbiology ; Phylogeny ; Sri Lanka ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia are a group of maternally inherited intracellular bacteria known to be widespread among arthropods. Infections with Wolbachia cause declines of host populations, and also induce host resistance to a wide range of pathogens. Over the past few decades, researchers were curious to use Wolbachia as a biological tool to control mosquito vectors. During the present study, assessment of the prevalence of Wolbachia infections among wild mosquito populations in Sri Lanka where mosquito-borne diseases are a major health concern, was carried out for the first time. DNA was extracted from the abdomens of mosquitoes, collected from seven provinces, and screened for the presence of Wolbachia by PCR using wsp and groE primers. Group-specific and strain-specific primers were used to classify Wolbachia into the supergroups A and B, and into the strains Mel, AlbA and Pip.

RESULTS: A total of 330 individual mosquitoes belonging to 22 species and 7 genera were screened. Eighty-seven mosquitoes (26.36%) belonging to four species (i.e. Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Mansonia uniformis) were positive for Wolbachia infections. Primary vector of the dengue fever, Ae. aegypti was negative for Wolbachia infections while the secondary vector, Ae. albopictus, showed a very high infection rate. The filarial vector C. quinquefasciatus had a relatively high rate of infection. Japanese encephalitis vectors C. gelidus and C. triteaneorynchus, and the Anopheles vectors of malaria were negative for Wolbachia infections. Nine sequences of Wolbachia-positive PCR products were deposited in the GenBank and compared with other available data. Aedes albopictus was infected with both Wolbachia strains A (AlbA) and B (Pip) supergroups. Phylogenetic analysis of the wsp sequences showed two major branches confirming identities obtained from the PCR screening with strain-specific primers.

CONCLUSION: Wolbachia infections were found only among four mosquito species in Sri Lanka: Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Mansonia uniformis. Sequence data showed high haplotype diversity among the Wolbachia strains.}, } @article {pmid28486424, year = {2017}, author = {Fenner, J and Seltzer, J and Peyton, S and Sullivan, H and Tolson, P and Walsh, RP and Hill, J and Counterman, BA}, title = {Demographic Variation of Wolbachia Infection in the Endangered Mitchell's Satyr Butterfly.}, journal = {Insects}, volume = {8}, number = {2}, pages = {}, pmid = {28486424}, issn = {2075-4450}, abstract = {The Mitchell's satyr, Neonympha mitchellii, is an endangered species that is limited to highly isolated habitats in the northern and southern United States. Conservation strategies for isolated endangered species often implement captive breeding and translocation programs for repopulation. However, these programs risk increasing the spread of harmful pathogens, such as the bacterial endosymbiont Wolbachia. Wolbachia can manipulate the host's reproduction leading to incompatibilities between infected and uninfected hosts. This study uses molecular methods to screen for Wolbachia presence across the distribution of the Mitchell's satyr and its subspecies, St. Francis satyr, which are both federally listed as endangered and are considered two of the rarest butterflies in North America. The screens confirmed the presence of Wolbachia in the northern and newly discovered southern populations of the Mitchell's satyr, but not in the St. Francis satyr population. These results combined with previous reports of Wolbachia in N. mitchellii, highlight that Wolbachia infection varies both geographically and temporally in satyr populations. The temporal variance shows the importance of continued monitoring of Wolbachia infection during conservation programs. To reduce the risk of reproductive incompatibilities, it is advised that all individuals collected for conservation purposes be screened for Wolbachia and recommended to avoid the use of infected individuals for captive breeding and translocation programs.}, } @article {pmid28477267, year = {2017}, author = {Toribio-Fernández, R and Bella, JL and Martínez-Rodríguez, P and Funkhouser-Jones, LJ and Bordenstein, SR and Pita, M}, title = {Chromosomal localization of Wolbachia inserts in the genomes of two subspecies of Chorthippus parallelus forming a Pyrenean hybrid zone.}, journal = {Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology}, volume = {25}, number = {3-4}, pages = {215-225}, pmid = {28477267}, issn = {1573-6849}, support = {P30 DK058404/DK/NIDDK NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; *Chromosomes, Bacterial ; *Genome, Insect ; Grasshoppers/*genetics ; Heterochromatin ; *Hybridization, Genetic ; In Situ Hybridization, Fluorescence ; Male ; *Mutagenesis, Insertional ; *Polytene Chromosomes ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {Wolbachia are endosymbiotic bacteria of arthropods and nematodes that can manipulate the reproduction of various host organisms to facilitate their own maternal transmission. Moreover, Wolbachia's presence in host germ cells may contribute to the many cases of lateral gene transfer from Wolbachia to host genomes that have been described. A previous study in Chorthippus parallelus, a well-known orthopteroid forming a hybrid zone in the Pyrenees, identified Wolbachia sequences from two major supergroups in the genomes of infected and uninfected Chorthippus parallelus parallelus (Cpp) and Chorthippus parallelus erythropus (Cpe) subspecies. In this study, we map the Wolbachia genomic inserts to specific regions on the chromosomes of Cpp and Cpe by fluorescent in situ hybridization (FISH) using tyramides to increase the accuracy and detection of these insertions. Additionally, we consider some of the possible roles that these bacterial inserts play in the organization and function of the grasshopper genome, as well as how they can serve as markers for phylogenetic relationships of these organisms.}, } @article {pmid28471518, year = {2017}, author = {Li, S and Zhou, C and Chen, G and Zhou, Y}, title = {Bacterial microbiota in small brown planthopper populations with different rice viruses.}, journal = {Journal of basic microbiology}, volume = {57}, number = {7}, pages = {590-596}, doi = {10.1002/jobm.201700004}, pmid = {28471518}, issn = {1521-4028}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Hemiptera/*microbiology/virology ; High-Throughput Nucleotide Sequencing ; Insect Vectors/*microbiology/virology ; *Microbiota/genetics ; Oryza ; Plant Viruses/isolation & purification/physiology ; RNA, Ribosomal, 16S ; *Symbiosis ; Tenuivirus/isolation & purification/physiology ; Wolbachia/genetics/isolation & purification ; }, abstract = {The small brown planthopper (SBPH) is an important virus vector, transmitting Rice stripe virus (RSV), and Rice black-streaked dwarf virus (RBSDV). Insect symbionts play an essential role in the insect fitness, however, it is still unclear about their contributions to viral transmission by SBPH. Here, we investigated endosymbiont communities in non-viruliferous, RSV-infected, and RBSDV-infected SBPH populations using Illumina 16S rRNA gene MiSeq sequencing. In total, 281,803 effective sequences of the 16S rRNA gene were generated from different samples. Sequence analysis revealed the percentages of these bacterial groups in different SBPH populations on several taxonomic levels ranging from phyla to genera. The extremely consistent bacterial diversity and abundance indicated that RSV or RBSDV infection did not affect the composition and abundance of symbionts in SBPH. It was notable that Wolbachia was dominant in all populations. The symbiosis between Wolbachia and SBPH might be potentially studied and utilized to control pest SBPH in the future.}, } @article {pmid28464440, year = {2017}, author = {Martinez, J and Tolosana, I and Ok, S and Smith, S and Snoeck, K and Day, JP and Jiggins, FM}, title = {Symbiont strain is the main determinant of variation in Wolbachia-mediated protection against viruses across Drosophila species.}, journal = {Molecular ecology}, volume = {26}, number = {15}, pages = {4072-4084}, pmid = {28464440}, issn = {1365-294X}, support = {//Wellcome Trust/United Kingdom ; 281668/ERC_/European Research Council/International ; }, mesh = {Animals ; *Disease Resistance ; Drosophila/genetics/*microbiology/virology ; Genome, Bacterial ; Genome, Insect ; Phenotype ; *Symbiosis ; Viruses/pathogenicity ; Wolbachia/*genetics ; }, abstract = {Wolbachia is a common heritable bacterial symbiont in insects. Its evolutionary success lies in the diverse phenotypic effects it has on its hosts coupled to its propensity to move between host species over evolutionary timescales. In a survey of natural host-symbiont associations in a range of Drosophila species, we found that 10 of 16 Wolbachia strains protected their hosts against viral infection. By moving Wolbachia strains between host species, we found that the symbiont genome had a much greater influence on the level of antiviral protection than the host genome. The reason for this was that the level of protection depended on the density of the symbiont in host tissues, and Wolbachia rather than the host-controlled density. The finding that virus resistance and symbiont density are largely under the control of symbiont genes in this system has important implications both for the evolution of these traits and for public health programmes using Wolbachia to prevent mosquitoes from transmitting disease.}, } @article {pmid28446677, year = {2017}, author = {Schultz, MJ and Isern, S and Michael, SF and Corley, RB and Connor, JH and Frydman, HM}, title = {Variable Inhibition of Zika Virus Replication by Different Wolbachia Strains in Mosquito Cell Cultures.}, journal = {Journal of virology}, volume = {91}, number = {14}, pages = {}, pmid = {28446677}, issn = {1098-5514}, support = {R01 AI099210/AI/NIAID NIH HHS/United States ; R21 NS101151/NS/NINDS NIH HHS/United States ; R56 AI097589/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes ; Animals ; *Antibiosis ; Cell Line ; Protein Biosynthesis ; RNA, Viral/biosynthesis ; Transcription, Genetic ; Viral Nonstructural Proteins/biosynthesis ; *Virus Replication ; Wolbachia/*physiology ; Zika Virus/*physiology ; }, abstract = {Mosquito-borne arboviruses are a major source of human disease. One strategy to reduce arbovirus disease is to reduce the mosquito's ability to transmit virus. Mosquito infection with the bacterial endosymbiont Wolbachia pipientis wMel is a novel strategy to reduce Aedes mosquito competency for flavivirus infection. However, experiments investigating cyclic environmental temperatures have shown a reduction in maternal transmission of wMel, potentially weakening the integration of this strain into a mosquito population relative to that of other Wolbachia strains. Consequently, it is important to investigate additional Wolbachia strains. All Zika virus (ZIKV) suppression studies are limited to the wMel Wolbachia strain. Here we show ZIKV inhibition by two different Wolbachia strains: wAlbB (isolated from Aedes albopictus mosquitoes) and wStri (isolated from the planthopper Laodelphax striatellus) in mosquito cells. Wolbachia strain wStri inhibited ZIKV most effectively. Single-cycle infection experiments showed that ZIKV RNA replication and nonstructural protein 5 translation were reduced below the limits of detection in wStri-containing cells, demonstrating early inhibition of virus replication. ZIKV replication was rescued when Wolbachia was inhibited with a bacteriostatic antibiotic. We observed a partial rescue of ZIKV growth when Wolbachia-infected cells were supplemented with cholesterol-lipid concentrate, suggesting competition for nutrients as one of the possible mechanisms of Wolbachia inhibition of ZIKV. Our data show that wAlbB and wStri infection causes inhibition of ZIKV, making them attractive candidates for further in vitro mechanistic and in vivo studies and future vector-centered approaches to limit ZIKV infection and spread.IMPORTANCE Zika virus (ZIKV) has swiftly spread throughout most of the Western Hemisphere. This is due in large part to its replication in and spread by a mosquito vector host. There is an urgent need for approaches that limit ZIKV replication in mosquitoes. One exciting approach for this is to use a bacterial endosymbiont called Wolbachia that can populate mosquito cells and inhibit ZIKV replication. Here we show that two different strains of Wolbachia, wAlbB and wStri, are effective at repressing ZIKV in mosquito cell lines. Repression of virus growth is through the inhibition of an early stage of infection and requires actively replicating Wolbachia Our findings further the understanding of Wolbachia viral inhibition and provide novel tools that can be used in an effort to limit ZIKV replication in the mosquito vector, thereby interrupting the transmission and spread of the virus.}, } @article {pmid28439127, year = {2017}, author = {Becking, T and Giraud, I and Raimond, M and Moumen, B and Chandler, C and Cordaux, R and Gilbert, C}, title = {Diversity and evolution of sex determination systems in terrestrial isopods.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {1084}, pmid = {28439127}, issn = {2045-2322}, mesh = {Animals ; *Evolution, Molecular ; Gene Expression Profiling ; *Genetic Variation ; Isopoda/classification/*genetics ; Phylogeny ; *Sex Characteristics ; }, abstract = {Sex determination systems are highly variable in many taxa, sometimes even between closely related species. Yet the number and direction of transitions between these systems have seldom been characterized, and the underlying mechanisms are still poorly understood. Here we generated transcriptomes for 19 species of terrestrial isopod crustaceans, many of which are infected by Wolbachia bacterial endosymbionts. Using 88 single-copy orthologous genes, we reconstructed a fully resolved and dated phylogeny of terrestrial isopods. An original approach involving crossings of sex-reversed individuals allowed us to characterize the heterogametic systems of five species (one XY/XX and four ZW/ZZ). Mapping of these and previously known heterogametic systems onto the terrestrial isopod phylogeny revealed between 3 and 13 transitions of sex determination systems during the evolution of these taxa, most frequently from female to male heterogamety. Our results support that WW individuals are viable in many species, suggesting sex chromosomes are at an incipient stage of their evolution. Together, these data are consistent with the hypothesis that nucleo-cytoplasmic conflicts generated by Wolbachia endosymbionts triggered recurrent turnovers of sex determination systems in terrestrial isopods. They further establish terrestrial isopods as a model to study evolutionary transitions in sex determination systems and pave the way to molecularly characterize these systems.}, } @article {pmid28435138, year = {2017}, author = {Baldridge, G and Higgins, L and Witthuhn, B and Markowski, T and Baldridge, A and Armien, A and Fallon, A}, title = {Proteomic analysis of a mosquito host cell response to persistent Wolbachia infection.}, journal = {Research in microbiology}, volume = {168}, number = {7}, pages = {609-625}, pmid = {28435138}, issn = {1769-7123}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/metabolism/*microbiology/virology ; Amino Acids/metabolism ; Animals ; Carbohydrate Metabolism ; Cell Line ; Cytoskeletal Proteins ; Energy Metabolism ; Flavivirus/genetics/physiology ; Insect Proteins/genetics/*metabolism ; Lipid Metabolism ; Membrane Proteins/metabolism ; Nucleotides/metabolism ; Proteome ; Proteomics ; Secondary Metabolism ; Signal Transduction ; Wolbachia/isolation & purification/*metabolism/virology ; }, abstract = {Wolbachia pipientis, an obligate intracellular bacterium associated with arthropods and filarial worms, is a target for filarial disease treatment and provides a gene drive agent for insect vector population suppression/replacement. We compared proteomes of Aedes albopictus mosquito C/wStr1 cells persistently infected with Wolbachia strain wStr, relative to uninfected C7-10 control cells. Among approximately 2500 proteins, iTRAQ data identified 815 differentially abundant proteins. As functional classes, energy and central intermediary metabolism proteins were elevated in infected cells, while suppressed proteins with roles in host DNA replication, transcription and translation suggested that Wolbachia suppresses pathways that support host cell growth and proliferation. Vacuolar ATPase subunits were strongly elevated, consistent with high densities of Wolbachia contained individually within vacuoles. Other differential level proteins had roles in ROS neutralization, protein modification/degradation and signaling, including hypothetical proteins whose functions in Wolbachia infection can potentially be manipulated by RNAi interference or transfection. Detection of flavivirus proteins supports further analysis of poorly understood, insect-specific flaviviruses and their potential interactions with Wolbachia, particularly in mosquitoes transinfected with Wolbachia. This study provides a framework for future attempts to manipulate pathways in insect cell lines that favor production of Wolbachia for eventual genetic manipulation, transformation and transinfection of vector species.}, } @article {pmid28426794, year = {2017}, author = {Monnin, D and Kremer, N and Desouhant, E and Vavre, F}, title = {Impact of Wolbachia on oxidative stress sensitivity in the parasitic wasp Asobara japonica.}, journal = {PloS one}, volume = {12}, number = {4}, pages = {e0175974}, pmid = {28426794}, issn = {1932-6203}, mesh = {Animals ; Host-Pathogen Interactions ; *Oxidative Stress ; Symbiosis ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {The oxidative homeostasis is the balance between reactive oxygen species and antioxidant molecules. In addition to be considered as a key factor underlying life-history traits evolution, the oxidative homeostasis has been shown to be involved in many host-symbiont associations. Previous studies suggest an interaction between the bacterial endosymbiont Wolbachia and the oxidative homeostasis of some insect hosts. This interaction is likely to exert a strong influence on the host evolution, as it has been proposed in the wasp Asobara tabida, whose dependence upon Wolbachia is due to the evolutionary loss of its ability to regulate the oxidative homeostasis in the absence of the symbiont. Although such cases of complete dependence are rare, cases of insects having lost only a part of their autonomy over the control of the oxidative homeostasis might be more common. If so, one can expect that insects having coevolved with Wolbachia will be more sensitive to oxidative stress when cured of their symbionts. We tested this hypothesis by studying the effects of an experimentally-induced oxidative stress on various life-history traits of Asobara japonica, a species closely related to A. tabida. For most of the life-history traits studied, the sensitivity of the wasps to oxidative stress did not correlate with their infection status. The only exception was the parasitic success. However, contrarily to our expectation, the sensitivity to oxidative stress was increased, rather than decreased, when Wolbachia was present. This result suggests that Wolbachia does not participate to mitigate oxidative stress in A. japonica, and that on the contrary its presence might still be costly in stressful environments.}, } @article {pmid28421042, year = {2017}, author = {Novakova, E and Woodhams, DC and Rodríguez-Ruano, SM and Brucker, RM and Leff, JW and Maharaj, A and Amir, A and Knight, R and Scott, J}, title = {Mosquito Microbiome Dynamics, a Background for Prevalence and Seasonality of West Nile Virus.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {526}, pmid = {28421042}, issn = {1664-302X}, abstract = {Symbiotic microbial communities augment host phenotype, including defense against pathogen carriage and infection. We sampled the microbial communities in 11 adult mosquito host species from six regions in southern Ontario, Canada over 3 years. Of the factors examined, we found that mosquito species was the largest driver of the microbiota, with remarkable phylosymbiosis between host and microbiota. Seasonal shifts of the microbiome were consistently repeated over the 3-year period, while region had little impact. Both host species and seasonal shifts in microbiota were associated with patterns of West Nile virus (WNV) in these mosquitoes. The highest prevalence of WNV, with a seasonal spike each year in August, was in the Culex pipiens/restuans complex, and high WNV prevalence followed a decrease in relative abundance of Wolbachia in this species. Indeed, mean temperature, but not precipitation, was significantly correlated with Wolbachia abundance. This suggests that at higher temperatures Wolbachia abundance is reduced leading to greater susceptibility to WNV in the subsequent generation of C. pipiens/restuans hosts. Different mosquito genera harbored significantly different bacterial communities, and presence or abundance of Wolbachia was primarily associated with these differences. We identified several operational taxonomic units (OTUs) of Wolbachia that drive overall microbial community differentiation among mosquito taxa, locations and timepoints. Distinct Wolbachia OTUs were consistently found to dominate microbiomes of Cx. pipiens/restuans, and of Coquilletidia perturbans. Seasonal fluctuations of several other microbial taxa included Bacillus cereus, Enterococcus, Methylobacterium, Asaia, Pantoea, Acinetobacter johnsonii, Pseudomonas, and Mycoplasma. This suggests that microbiota may explain some of the variation in vector competence previously attributed to local environmental processes, especially because Wolbachia is known to affect carriage of viral pathogens.}, } @article {pmid28420428, year = {2017}, author = {Armoo, S and Doyle, SR and Osei-Atweneboana, MY and Grant, WN}, title = {Significant heterogeneity in Wolbachia copy number within and between populations of Onchocerca volvulus.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {188}, pmid = {28420428}, issn = {1756-3305}, mesh = {Animals ; *DNA Copy Number Variations ; Ecotype ; Forests ; *Genetic Heterogeneity ; *Genome, Bacterial ; Genome, Helminth ; Grassland ; High-Throughput Nucleotide Sequencing ; Ivermectin/pharmacology ; Onchocerca volvulus/genetics/*microbiology ; Real-Time Polymerase Chain Reaction ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Wolbachia are intracellular bacteria found in arthropods and several filarial nematode species. The filarial Wolbachia have been proposed to be involved in the immunopathology associated with onchocerciasis. Higher Wolbachia-to-nematode ratios have been reported in the savannah-ecotype compared to the forest-ecotype, and have been interpreted as consistent with a correlation between Wolbachia density and disease severity. However, factors such as geographic stratification and ivermectin drug exposure can lead to significant genetic heterogeneity in the nematode host populations, so we investigated whether Wolbachia copy number variation is also associated with these underlying factors.

METHODS: Genomic DNA was prepared from single adult nematodes representing forest and savannah ecotypes sampled from Togo, Ghana, Côte d'Ivoire and Mali. A qPCR assay was developed to measure the number of Wolbachia genome(s) per nematode genome. Next-generation sequencing (NGS) was also used to measure relative Wolbachia copy number, and independently verify the qPCR assay.

RESULTS: Significant variation was observed within the forest (range: 0.02 to 452.99; median: 10.58) and savannah (range: 0.01 to 1106.25; median: 9.10) ecotypes, however, no significant difference between ecotypes (P = 0.645) was observed; rather, strongly significant Wolbachia variation was observed within and between the nine study communities analysed (P = 0.021), independent of ecotype. Analysis of ivermectin-treated and untreated nematodes by qPCR showed no correlation (P = 0.869); however, an additional analysis of a subset of the nematodes by qPCR and NGS revealed a correlation between response to ivermectin treatment and Wolbachia copy number (P = 0.020).

CONCLUSIONS: This study demonstrates that extensive within and between population variation exists in the Wolbachia content of individual adult O. volvulus. The origin and functional significance of such variation (up to ~ 100,000-fold between worms; ~10 to 100-fold between communities) in the context of the proposed mutualistic relationship between the worms and the bacteria, and between the presence of Wolbachia and clinical outcome of infection, remains unclear. These data do not support a correlation between Wolbachia copy number and forest or savannah ecotype, and may have implications for the development of anti-Wolbachia drugs as a macrofilaricidal treatment of onchocerciasis. The biological significance of a correlation between variation in Wolbachia copy number and ivermectin response remains unexplained.}, } @article {pmid28419609, year = {2017}, author = {Russell, JA and Oliver, KM and Hansen, AK}, title = {Band-aids for Buchnera and B vitamins for all.}, journal = {Molecular ecology}, volume = {26}, number = {8}, pages = {2199-2203}, doi = {10.1111/mec.14047}, pmid = {28419609}, issn = {1365-294X}, mesh = {Animals ; Aphids/genetics ; Buchnera/*genetics ; Phylogeny ; Symbiosis/genetics ; *Vitamin B Complex ; }, abstract = {Evolution lacks foresight, and hence, key adaptations may produce major challenges over the long run. The natural world is rife with examples of long-term 'side effects' associated with quick-fix tinkering, including blind spots in vertebrate eyes. An important question is how nature compensates for imperfections once evolution has set a course. The symbioses associated with sap-feeding insects present a fascinating opportunity to address this issue. On one hand, the substantial diversity and biomass of sap-feeding insects are largely due to ancient acquisitions of nutrient-provisioning bacterial symbionts. Yet, the insularity and small population sizes enforced by intracellular life and strict maternal transfer inevitably result in the degradation of symbiont genomes and, often, the beneficial services that symbionts provide. Stabilization through lateral transfer of bacterial genes into the host nucleus (often from exogenous sources) or replacement of the long-standing symbiont with a new partner are potential solutions to this evolutionary dilemma (Bennett & Moran). A third solution is adoption of a cosymbiont that compensates for specific losses in the original resident. Ancient 'co-obligate' symbiont pairs in mealybugs, leafhoppers, cicadas and spittlebugs show colocalization, codiversification, metabolite exchange and generally nonredundant nutrient biosynthesis (Bennett & Moran). But in this issue, Meseguer et al. () report on a different flavour of cosymbiosis among conifer-feeding Cinara aphids.}, } @article {pmid28412513, year = {2017}, author = {Ju, JF and Hoffmann, AA and Zhang, YK and Duan, XZ and Guo, Y and Gong, JT and Zhu, WC and Hong, XY}, title = {Wolbachia-induced loss of male fertility is likely related to branch chain amino acid biosynthesis and iLvE in Laodelphax striatellus.}, journal = {Insect biochemistry and molecular biology}, volume = {85}, number = {}, pages = {11-20}, doi = {10.1016/j.ibmb.2017.04.002}, pmid = {28412513}, issn = {1879-0240}, mesh = {Amino Acids/*biosynthesis ; Animals ; Copulation ; Female ; Fertility ; Hemiptera/metabolism/*microbiology ; *Host-Pathogen Interactions ; Insect Proteins/metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; Testis/metabolism ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbionts that infect many species of arthropods and nematodes. Wolbachia-induced cytoplasmic incompatibility (CI) is the most common phenotype in affected hosts, involving embryonic lethality in crosses between Wolbachia-infected males and uninfected females. The molecular mechanisms underlying this phenomenon are currently unclear. Here we examine the molecular correlates of the Wolbachia infection in Laodelphax striatellus (Fallén), an important rice pest, where embryonic lethality is strong and almost complete. We compared the gene expression of 4-day-old Wolbachia-infected and uninfected L. striatellus testes to identify candidate genes for paternal-effect embryonic lethality induction. Based on microarray analysis, iLvE was the most down-regulated gene; this gene mediates branched-chain amino acid (BCAA) biosynthesis and participates in many processes related to reproductive performance. After knocking down iLvE by RNAi in uninfected male L. striatellus, male fertility was reduced, leading to a decrease in embryo hatching rates, but fertility was rescued in crosses between these males and Wolbachia-infected females. Removal of BCAA in chemically-defined diets of uninfected males also led to a loss of male fertility. Low amino acid nutrition may enhance exposure time of sperm to Wolbachia in the testes to affect adult reproduction in L. striatellus by reducing the number of sperm transferred per mating by males. These results indicate that Wolbachia may decrease male fertility in L. striatellus by acting on iLvE, a key factor of BCAA biosynthesis, and delaying sperm maturation.}, } @article {pmid28411063, year = {2017}, author = {Turelli, M and Barton, NH}, title = {Deploying dengue-suppressing Wolbachia : Robust models predict slow but effective spatial spread in Aedes aegypti.}, journal = {Theoretical population biology}, volume = {115}, number = {}, pages = {45-60}, pmid = {28411063}, issn = {1096-0325}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/growth & development/*microbiology/physiology/virology ; Animals ; *Biological Control Agents ; Dengue/*prevention & control/transmission/virology ; Dengue Virus/growth & development/isolation & purification/*physiology ; Disease Vectors ; Female ; Humans ; Infection Control ; Male ; *Models, Biological ; Spatio-Temporal Analysis ; Wolbachia/growth & development/isolation & purification/*physiology ; }, abstract = {A novel strategy for controlling the spread of arboviral diseases such as dengue, Zika and chikungunya is to transform mosquito populations with virus-suppressing Wolbachia. In general, Wolbachia transinfected into mosquitoes induce fitness costs through lower viability or fecundity. These maternally inherited bacteria also produce a frequency-dependent advantage for infected females by inducing cytoplasmic incompatibility (CI), which kills the embryos produced by uninfected females mated to infected males. These competing effects, a frequency-dependent advantage and frequency-independent costs, produce bistable Wolbachia frequency dynamics. Above a threshold frequency, denoted pˆ, CI drives fitness-decreasing Wolbachia transinfections through local populations; but below pˆ, infection frequencies tend to decline to zero. If pˆ is not too high, CI also drives spatial spread once infections become established over sufficiently large areas. We illustrate how simple models provide testable predictions concerning the spatial and temporal dynamics of Wolbachia introductions, focusing on rate of spatial spread, the shape of spreading waves, and the conditions for initiating spread from local introductions. First, we consider the robustness of diffusion-based predictions to incorporating two important features of wMel-Aedes aegypti biology that may be inconsistent with the diffusion approximations, namely fast local dynamics induced by complete CI (i.e., all embryos produced from incompatible crosses die) and long-tailed, non-Gaussian dispersal. With complete CI, our numerical analyses show that long-tailed dispersal changes wave-width predictions only slightly; but it can significantly reduce wave speed relative to the diffusion prediction; it also allows smaller local introductions to initiate spatial spread. Second, we use approximations for pˆ and dispersal distances to predict the outcome of 2013 releases of wMel-infected Aedes aegypti in Cairns, Australia, Third, we describe new data from Ae. aegypti populations near Cairns, Australia that demonstrate long-distance dispersal and provide an approximate lower bound on pˆ for wMel in northeastern Australia. Finally, we apply our analyses to produce operational guidelines for efficient transformation of vector populations over large areas. We demonstrate that even very slow spatial spread, on the order of 10-20 m/month (as predicted), can produce area-wide population transformation within a few years following initial releases covering about 20-30% of the target area.}, } @article {pmid28405268, year = {2017}, author = {Ma, Y and Chen, WJ and Li, ZH and Zhang, F and Gao, Y and Luan, YX}, title = {Revisiting the phylogeny of Wolbachia in Collembola.}, journal = {Ecology and evolution}, volume = {7}, number = {7}, pages = {2009-2017}, pmid = {28405268}, issn = {2045-7758}, abstract = {The endosymbiont Wolbachia has been detected in a few parthenogenetic collembolans sampled in Europe and America, including three species of Poduromorpha, two species of Entomobryomorpha, and two species of Neelipleona. Based on 16S rRNA and ftsZ gene sequences, most of the Wolbachia infecting parthenogenetic collembolans were characterized as members of supergroup E and showed concordant phylogeny with their hosts. However, the two neelipleonan symbionts form another unique group, indicating that Wolbachia has infected parthenogenetic collembolans multiple times. In this study, five parthenogenetic collembolan species were identified as hosts of Wolbachia, and four new Wolbachia strains were reported for four collembolan species sampled in China, respectively, including a neelipleonan strain from Megalothorax incertus (wMinc). Our results demonstrated that the Wolbachia multilocus sequence typing (MLST) system is superior to the 16S rRNA + ftsZ approach for phylogenetic analyses of collembolan Wolbachia. The MLST system assigned these Wolbachia of parthenogenetic collembolans to supergroup E as a unique clade, which included wMinc, supporting the monophyletic origin of Wolbachia in parthenogenetic collembolan species. Moreover, our data suggested supergroup E as one of the most divergent lineages in Wolbachia and revealed the discrepancy between the phylogenies of Wolbachia from parthenogenetic collembolans and their hosts, which may result from the high level of genetic divergence between collembolan Wolbachia, in association with the geographic differentiation of their hosts, or the possible horizontal transmission of Wolbachia between different collembolan species.}, } @article {pmid28405008, year = {2017}, author = {Dunning Hotopp, JC and Slatko, BE and Foster, JM}, title = {Targeted Enrichment and Sequencing of Recent Endosymbiont-Host Lateral Gene Transfers.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {857}, pmid = {28405008}, issn = {2045-2322}, support = {DP2 OD007372/OD/NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Genome, Helminth ; Nematoda/genetics/microbiology ; *Symbiosis ; Wolbachia/genetics/pathogenicity ; }, abstract = {Lateral gene transfer (LGT) from microbial symbionts to invertebrate animals is described at an increasing rate, particularly between Wolbachia endosymbionts and their diverse invertebrate hosts. We sought to assess the use of a capture system to cost-effectively sequence such LGT from the host genome. The sequencing depth of Illumina paired end data obtained with a Wolbachia capture system correlated well with that for an Illumina paired end data set used to detect LGT in Wolbachia-depleted B. malayi (p-value: <2e-16). Using a sequencing depth threshold of two or three standard deviations above the mean, 96.9% or 96.7% of positions, respectively, are predicted in the same manner between the two datasets, with 24.7% or 42.5% of the known 49.0 kbp of LGT sequence predicted correctly, respectively. Prior qPCR results for nuwts showed similar correlations for both datasets supporting our conclusion that oligonucleotide-based capture methods can be used to obtain sequences from Wolbachia-host LGT. However, at least 121 positions had a minority of the reads supporting the endosymbiont reference base call using the capture data, illustrating that sequence reads from endosymbiont-host LGTs can confound endosymbiont genome projects, erroneously altering the called consensus genome, a problem that is irrespective to the sequencing technology or platform.}, } @article {pmid28403439, year = {2017}, author = {Ritchie, SA and Johnson, BJ}, title = {Advances in Vector Control Science: Rear-and-Release Strategies Show Promise… but Don't Forget the Basics.}, journal = {The Journal of infectious diseases}, volume = {215}, number = {suppl_2}, pages = {S103-S108}, doi = {10.1093/infdis/jiw575}, pmid = {28403439}, issn = {1537-6613}, mesh = {Aedes/*physiology/virology ; Animals ; Chikungunya Fever/*prevention & control/transmission ; Dengue/*prevention & control/transmission ; Humans ; Insect Vectors/*physiology/virology ; *Mosquito Control ; Reproduction ; }, abstract = {Both chikungunya and Zika viruses have recently swept from Africa across the Pacific to the Americas, causing major outbreaks of disease in humans. In the meantime, dengue epidemics continue throughout the tropics. Traditional vector control programs based on strategies from 1950s and 1960s have been relatively ineffective in combating recent epidemics. In response, new methods involving the rearing and releasing of large numbers of mosquitoes to eliminate or modify local Aedes populations are being developed, with several currently conducting field releases in high-risk countries. These advances, include the release of Wolbachia-infected Aedes aegypti and Aedes albopictus, for either its virus-blocking capabilities, sterilization by cytoplasmic incompatibility, or both; the release of Aedes carrying dominant lethal genes, such as the OX513A strain of A. aegypti; and other emerging techniques, such as advancing gene-drive technologies, are summarized, as well as current stages of development and primary operational and regulatory hurdles. Although these technologies show great promise, none are ready for widespread rollout for cities of millions of people. Thus, efforts should be made to avoid methods such as space sprays that have failed and improve existing technologies to increase their efficacy.}, } @article {pmid28403227, year = {2017}, author = {Sumi, T and Miura, K and Miyatake, T}, title = {Wolbachia density changes seasonally amongst populations of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae).}, journal = {PloS one}, volume = {12}, number = {4}, pages = {e0175373}, pmid = {28403227}, issn = {1932-6203}, mesh = {Animals ; Butterflies/genetics/*microbiology/physiology ; DNA, Mitochondrial/genetics ; Female ; Haplotypes ; *Host-Pathogen Interactions ; Japan ; Male ; Phylogeny ; Reproduction ; Rickettsiaceae Infections/veterinary ; Seasons ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Previous studies showed that the survival rate of Wolbachia decreases under high temperature in incubators. It is also known that a high density of Wolbachia in the host body reduces the host emergence rate, while low densities fail to change reproduction rates. However, few studies have examined the density of Wolbachia in hosts in the field. Here, we focus on Wolbachia infection of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae), which is distributed throughout the Japanese islands. We examined the rate and density of Wolbachia infection in the bodies of butterflies at thirteen locations in Japan. At seven of these places, we collected butterflies in different seasons to determine seasonal differences in the infection rate and density and found that Wolbachia density has seasonal differences within the same population. Moreover, to determine whether Wolbachia density has a geographical cline, we compared the infection density of Wolbachia amongst all geographical populations. In addition, we determined the sequences of Wolbachia wsp and host mtDNA CO1 haplotypes of all populations. The results showed that Wolbachia density increased in early summer and decreased in autumn. Further, the density of Wolbachia infecting the same strain of Z. maha varied amongst populations, although no tendency in geographical cline was observed.}, } @article {pmid28401481, year = {2017}, author = {Carvalho, FD and Moreira, LA}, title = {Why is Aedes aegypti Linnaeus so Successful as a Species?.}, journal = {Neotropical entomology}, volume = {46}, number = {3}, pages = {243-255}, pmid = {28401481}, issn = {1678-8052}, mesh = {*Aedes/microbiology/virology ; Animals ; Brazil ; Humans ; *Insect Vectors ; Insecticides ; Population Dynamics ; Wolbachia ; Zika Virus ; Zika Virus Infection ; }, abstract = {Diseases transmitted by mosquitoes impose enormous burden towards human morbidity and mortality. Over the last three decades, Brazil has suffered from severe Dengue epidemics. In September 2014, this situation is further complicated by the introduction of two other viruses, Zika and Chikungunya, placing Brazil in a triple epidemic. In this article, we discuss the biology of Aedes aegypti Linnaeus, and the principal initiatives currently used to control mosquito populations and the diseases they transmit. Aedes aegypti has broad global distribution and is involved in the transmission of various arboviral diseases such as Dengue, Zika, and Chikungunya. Several factors contribute to the success of the species, particularly behavioral plasticity, rapid development, desiccation-resistant eggs, resistance to the principle insecticide classes currently available on the market, preference for the urban environment, and proximity to humans. Vector control programs are the best way to reduce the burden of mosquito-borne diseases. Chemical control is most commonly used in recent times, and unfortunately, the results have not been satisfactory but instead, there is increased vector dispersal and, subsequently, the spread of disease epidemics. Investigations of alternative control methods such as release of Wolbachia-infected mosquitoes for blocking vector-borne pathogens, release of transgenic mosquitoes carrying a lethal gene for offspring, and the use of insecticide-dispersing mosquitoes are under way in Brazil, and some have shown promising results. Special emphasis should be placed on integrated management of all available tactics, so as to maximize efforts towards mosquito control. Finally, we emphasize that continuous actions and community participation control initiatives are critically important for success.}, } @article {pmid28386769, year = {2017}, author = {Fromont, C and Riegler, M and Cook, JM}, title = {Relative Abundance and Strain Diversity in the Bacterial Endosymbiont Community of a Sap-Feeding Insect Across Its Native and Introduced Geographic Range.}, journal = {Microbial ecology}, volume = {74}, number = {3}, pages = {722-734}, pmid = {28386769}, issn = {1432-184X}, support = {HSF 12/10//The Hermon Slade Foundation/ ; }, mesh = {Animals ; Australia ; Bacteria/*classification/genetics ; *Bacterial Physiological Phenomena ; Female ; Ficus/growth & development ; Hemiptera/growth & development/*microbiology ; Herbivory ; High-Throughput Nucleotide Sequencing ; Male ; *Microbiota ; New Zealand ; Nymph/growth & development/microbiology ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; }, abstract = {Most insects are associated with bacterial symbionts. The bacterial diversity and community composition within hosts may play an important role in shaping insect population biology, ecology and evolution. We focussed on the bacterial microbiome of the Australian fig homotomid Mycopsylla fici (Hemiptera: Psylloidea), which can cause defoliation of its only host tree, Ficus macrophylla. This sap-feeding insect is native to mainland Australia and Lord Howe Island (LHI) but also occurs where its host has been planted, notably in New Zealand. By using a high-throughput 16S rDNA amplicon sequencing approach, we compared the bacterial diversity and community composition in individual adult males of four host populations, Sydney, Brisbane, LHI and Auckland. We also compared males, females and nymphs of the Sydney population. The microbiome of M. fici was simple and consisted mostly of the following three maternally inherited endosymbiont species: the primary endosymbiont Carsonella, a secondary (S-) endosymbiont and Wolbachia. However, the relative abundance of their sequence reads varied between host populations, except for similarities between Sydney and Auckland. In addition, insects from Sydney and Auckland had identical bacterial strains supporting the hypothesis that Sydney is the source population for Auckland. In contrast, mainland and LHI populations harboured the same S-endosymbiont, co-diverged Carsonella but different Wolbachia strains. Besides detecting endosymbiont-specific patterns of either co-evolution or horizontal acquisition, our study highlights that relative abundance of maternally inherited endosymbionts should also be taken into account when studying bacterial communities across host populations, as variations in bacterial density may impact host biology and ecology.}, } @article {pmid28386418, year = {2017}, author = {Ramsey, JS and Chavez, JD and Johnson, R and Hosseinzadeh, S and Mahoney, JE and Mohr, JP and Robison, F and Zhong, X and Hall, DG and MacCoss, M and Bruce, J and Cilia, M}, title = {Protein interaction networks at the host-microbe interface in Diaphorina citri, the insect vector of the citrus greening pathogen.}, journal = {Royal Society open science}, volume = {4}, number = {2}, pages = {160545}, pmid = {28386418}, issn = {2054-5703}, abstract = {The Asian citrus psyllid (Diaphorina citri) is the insect vector responsible for the worldwide spread of 'Candidatus Liberibacter asiaticus' (CLas), the bacterial pathogen associated with citrus greening disease. Developmental changes in the insect vector impact pathogen transmission, such that D. citri transmission of CLas is more efficient when bacteria are acquired by nymphs when compared with adults. We hypothesize that expression changes in the D. citri immune system and commensal microbiota occur during development and regulate vector competency. In support of this hypothesis, more proteins, with greater fold changes, were differentially expressed in response to CLas in adults when compared with nymphs, including insect proteins involved in bacterial adhesion and immunity. Compared with nymphs, adult insects had a higher titre of CLas and the bacterial endosymbionts Wolbachia, Profftella and Carsonella. All Wolbachia and Profftella proteins differentially expressed between nymphs and adults are upregulated in adults, while most differentially expressed Carsonella proteins are upregulated in nymphs. Discovery of protein interaction networks has broad applicability to the study of host-microbe relationships. Using protein interaction reporter technology, a D. citri haemocyanin protein highly upregulated in response to CLas was found to physically interact with the CLas coenzyme A (CoA) biosynthesis enzyme phosphopantothenoylcysteine synthetase/decarboxylase. CLas pantothenate kinase, which catalyses the rate-limiting step of CoA biosynthesis, was found to interact with a D. citri myosin protein. Two Carsonella enzymes involved in histidine and tryptophan biosynthesis were found to physically interact with D. citri proteins. These co-evolved protein interaction networks at the host-microbe interface are highly specific targets for controlling the insect vector responsible for the spread of citrus greening.}, } @article {pmid28381207, year = {2017}, author = {Ramalho, MO and Bueno, OC and Moreau, CS}, title = {Microbial composition of spiny ants (Hymenoptera: Formicidae: Polyrhachis) across their geographic range.}, journal = {BMC evolutionary biology}, volume = {17}, number = {1}, pages = {96}, pmid = {28381207}, issn = {1471-2148}, mesh = {Animals ; Ants/genetics/*microbiology ; Bacteria/classification/genetics/*isolation & purification ; Biological Evolution ; DNA, Bacterial ; High-Throughput Nucleotide Sequencing ; *Microbiota ; Phylogeny ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {BACKGROUND: Symbiotic relationships between insects and bacteria are found across almost all insect orders, including Hymenoptera. However there are still many remaining questions about these associations including what factors drive host-associated bacterial composition. To better understand the evolutionary significance of this association in nature, further studies addressing a diversity of hosts across locations and evolutionary history are necessary. Ants of the genus Polyrhachis (spiny ants) are distributed across the Old World and exhibit generalist diets and habits. Using Next Generation Sequencing (NGS) and bioinformatics tools, this study explores the microbial community of >80 species of Polyrhachis distributed across the Old World and compares the microbiota of samples and related hosts across different biogeographic locations and in the context of their phylogenetic history.

RESULTS: The predominant bacteria across samples were Enterobacteriaceae (Blochmannia - with likely many new strains), followed by Wolbachia (with multiple strains), Lactobacillus, Thiotrichaceae, Acinetobacter, Nocardia, Sodalis, and others. We recovered some exclusive strains of Enterobacteriaceae as specific to some subgenera of Polyrhachis, corroborating the idea of coevolution between host and bacteria for this bacterial group. Our correlation results (partial mantel and mantel tests) found that host phylogeny can influence the overall bacterial community, but that geographic location had no effect.

CONCLUSIONS: Our work is revealing important aspects of the biology of hosts in structuring the diversity and abundance of these host-associated bacterial communities including the role of host phylogeny and shared evolutionary history.}, } @article {pmid28358880, year = {2017}, author = {Grote, A and Voronin, D and Ding, T and Twaddle, A and Unnasch, TR and Lustigman, S and Ghedin, E}, title = {Defining Brugia malayi and Wolbachia symbiosis by stage-specific dual RNA-seq.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {3}, pages = {e0005357}, pmid = {28358880}, issn = {1935-2735}, support = {R56 AI101372/AI/NIAID NIH HHS/United States ; R56 AI118936/AI/NIAID NIH HHS/United States ; T32 AI007180/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*growth & development/*microbiology ; Female ; *Gene Expression Profiling ; Sequence Analysis, RNA ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Filarial nematodes currently infect up to 54 million people worldwide, with millions more at risk for infection, representing the leading cause of disability in the developing world. Brugia malayi is one of the causative agents of lymphatic filariasis and remains the only human filarial parasite that can be maintained in small laboratory animals. Many filarial nematode species, including B. malayi, carry an obligate endosymbiont, the alpha-proteobacteria Wolbachia, which can be eliminated through antibiotic treatment. Elimination of the endosymbiont interferes with development, reproduction, and survival of the worms within the mamalian host, a clear indicator that the Wolbachia are crucial for survival of the parasite. Little is understood about the mechanism underlying this symbiosis.

To better understand the molecular interplay between these two organisms we profiled the transcriptomes of B. malayi and Wolbachia by dual RNA-seq across the life cycle of the parasite. This helped identify functional pathways involved in this essential symbiotic relationship provided by the co-expression of nematode and bacterial genes. We have identified significant stage-specific and gender-specific differential expression in Wolbachia during the nematode's development. For example, during female worm development we find that Wolbachia upregulate genes involved in ATP production and purine biosynthesis, as well as genes involved in the oxidative stress response.

CONCLUSIONS/ SIGNIFICANCE: This global transcriptional analysis has highlighted specific pathways to which both Wolbachia and B. malayi contribute concurrently over the life cycle of the parasite, paving the way for the development of novel intervention strategies.}, } @article {pmid28357366, year = {2016}, author = {Caragata, EP and Dutra, HL and Moreira, LA}, title = {Inhibition of Zika virus by Wolbachia in Aedes aegypti.}, journal = {Microbial cell (Graz, Austria)}, volume = {3}, number = {7}, pages = {293-295}, doi = {10.15698/mic2016.07.513}, pmid = {28357366}, issn = {2311-2638}, abstract = {Through association with cases of microcephaly in 2015, Zika virus (ZIKV) has transitioned from a relatively unknown mosquito-transmitted pathogen to a global health emergency, emphasizing the need to improve existing mosquito control programs to prevent future disease outbreaks. The response to Zika must involve a paradigm shift from traditional to novel methods of mosquito control, and according to the World Health Organization should incorporate the release of mosquitoes infected with the bacterial endosymbiont Wolbachiapipientis. In our recent paper [Dutra, HLC et al., Cell Host & Microbe 2016] we investigated the potential of Wolbachia infections in Aedes aegypti to restrict infection and transmission of Zika virus recently isolated in Brazil. Wolbachia is now well known for its ability to block or reduce infection with a variety of pathogens in different mosquito species including the dengue (DENV), yellow fever, and chikungunya viruses, and malaria-causing Plasmodium, and consequently has great potential to control mosquito-transmitted diseases across the globe. Our results demonstrated that the wMel Wolbachia strain in Brazilian Ae. aegypti is a strong inhibitor of ZIKV infection, and furthermore appears to prevent transmission of infectious viral particles in mosquito saliva, which highlights the bacterium's suitability for more widespread use in Zika control.}, } @article {pmid28344097, year = {2017}, author = {Lefoulon, E and Giannelli, A and Makepeace, BL and Mutafchiev, Y and Townson, S and Uni, S and Verocai, GG and Otranto, D and Martin, C}, title = {Whence river blindness? The domestication of mammals and host-parasite co-evolution in the nematode genus Onchocerca.}, journal = {International journal for parasitology}, volume = {47}, number = {8}, pages = {457-470}, doi = {10.1016/j.ijpara.2016.12.009}, pmid = {28344097}, issn = {1879-0135}, mesh = {Animals ; Animals, Domestic/genetics/*parasitology ; *Biological Coevolution ; Electron Transport Complex IV/genetics/metabolism ; Gene Expression Regulation, Enzymologic ; Humans ; Mammals/*parasitology ; Onchocerca/*genetics/*physiology ; }, abstract = {The genus Onchocerca includes 34 described species and represents one of the largest genera of the filarial nematodes within the family Onchocercidae. Representative members of this genus are mainly parasites of ungulates, with some exceptions such as Onchocerca lupi and Onchocerca volvulus, infecting carnivores and/or humans. For a long time, the evolutionary relationships amongst onchocercids remained poorly studied, as the systematics of this genus was impaired by the high morphological variability of species included in the taxon. Although some molecular phylogenies were developed, these studies were mainly focused on bovine Onchocerca spp. and O. volvulus, including assessments of Wolbachia endosymbionts. In the present study, we analysed 13 Onchocerca spp. from a larger host spectrum using a panel of seven different genes. Analysis of the coxI marker supports its usefulness for the identification of species within the genus. The evolutionary history of the genus has been herein revised by multi-gene phylogenies, presenting three strongly supported clades of Onchocerca spp. Analyses of co-evolutionary scenarios between Onchocerca and their vertebrate hosts underline the effect of domestication on Onchocerca speciation. Our study indicates that a host switch event occurred between Bovidae, Canidae and humans. Cophylogenetic analyses between Onchocerca and the endosymbiotic bacterium Wolbachia indicate the strongest co-evolutionary pattern ever registered within the filarial nematodes. Finally, this dataset indicates that the clade composed by O. lupi, Onchocerca gutturosa, Onchocerca lienalis, Onchocerca ochengi and O. volvulus derived from recent speciation.}, } @article {pmid28337184, year = {2017}, author = {Joshi, D and Pan, X and McFadden, MJ and Bevins, D and Liang, X and Lu, P and Thiem, S and Xi, Z}, title = {The Maternally Inheritable Wolbachia wAlbB Induces Refractoriness to Plasmodium berghei in Anopheles stephensi.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {366}, pmid = {28337184}, issn = {1664-302X}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; }, abstract = {The endosymbiont Wolbachia wAlbB induces refractoriness to Plasmodium falciparum in Anopheles stephensi, the primary mosquito vector of human malaria in the Middle East and South Asia. However, it remains unknown whether such refractoriness can be extended to other malaria species. In particular, it was reported that under very specific conditions, wAlbB can enhance Plasmodium infection in some hosts. Here, we measured the impact of wAlbB on the rodent malaria parasite Plasmodium berghei in A. stephensi by comparing the load of oocysts and sporozoites in midguts and salivary glands, respectively, between wAlbB-infected and -uninfected mosquitoes. To investigate whether wAlbB modulated mosquito immune defense against parasites, we compared the expression of the immune genes, which were previously reported to involve in antimalarial response, in both midguts and the remaining carcass tissues of mosquitoes. The stable association of wAlbB with A. stephensi resulted in reduction of parasites by more than half at the oocyst stage, and up to 91.8% at the sporzoite stage. The anti-plasmodium immune genes, including TEP1, LRIM1, Toll pathway gene Rel1 and the effector Defensin 1, were induced by wAlbB in different mosquito body tissues. These findings suggest that immune priming is a potential cause of wAlbB-mediated antimalarial response in A. stephensi. More importantly, no evidence was found for any enhancement of Plasmodium infection in A. stephensi stably infected with wAlbB. We discuss these findings with possible implementations of Wolbachia for malaria control in disease endemic areas.}, } @article {pmid28334388, year = {2017}, author = {Cooper, WR and Garczynski, SF and Horton, DR and Unruh, TR and Beers, EH and Peter, WS and Hilton, RJ}, title = {Bacterial Endosymbionts of the Psyllid Cacopsylla pyricola (Hemiptera: Psyllidae) in the Pacific Northwestern United States.}, journal = {Environmental entomology}, volume = {46}, number = {2}, pages = {393-402}, doi = {10.1093/ee/nvx031}, pmid = {28334388}, issn = {1938-2936}, mesh = {Animals ; Bacteria/classification/genetics ; *Bacterial Physiological Phenomena ; Female ; Hemiptera/*microbiology ; Male ; Oregon ; Pyrus/growth & development ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Seasons ; Sequence Analysis, RNA ; *Symbiosis ; Washington ; }, abstract = {Insects often have facultative associations with bacterial endosymbionts, which can alter the insects' susceptibility to parasitism, pathogens, plant defenses, and certain classes of insecticides. We collected pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae), from pear orchards in Washington and Oregon, and surveyed them for the presence of bacterial endosymbionts. Adult psyllids were collected on multiple dates to allow us to assay specimens of both the summer ("summerform") and the overwintering ("winterform") morphotypes. Two endosymbionts, Arsenophonus and Phytoplasma pyri, were detected in psyllids of both morphotypes in both states. A separate survey revealed similar associations present in psyllids collected in 1987. Arsenophonus was present in 80-100% of psyllids in all growing regions. A slightly lower proportion of summerform than winterform psyllids harbored the bacterium. Arsenophonus was present in the bacteriomes and developing oocytes of most psyllids, indicating that this endosymbiont is transovarially transmitted. This bacterium was also observed in the salivary glands and midguts of some psyllids. Phytoplasma pyri was present in a greater proportion of pear psylla from orchards near Yakima, WA, than from other regions, and was present in a higher proportion of winterforms than summerforms. We did not detect Wolbachia, Profftella, or Liberibacter europaeus, which are associated with other psyllid pests, including other species of Cacopsylla. Our study is the first to survey North American populations of C. pyricola for endosymbionts, and provides a foundation for further research on how bacterial associations may influence the ecology and management of this pest.}, } @article {pmid28334194, year = {2017}, author = {Clarke, EL and Sundararaman, SA and Seifert, SN and Bushman, FD and Hahn, BH and Brisson, D}, title = {swga: a primer design toolkit for selective whole genome amplification.}, journal = {Bioinformatics (Oxford, England)}, volume = {33}, number = {14}, pages = {2071-2077}, pmid = {28334194}, issn = {1367-4811}, support = {R01 HL113252/HL/NHLBI NIH HHS/United States ; R01 HL087115/HL/NHLBI NIH HHS/United States ; R37 AI050529/AI/NIAID NIH HHS/United States ; T32 AI007532/AI/NIAID NIH HHS/United States ; P30 AI045008/AI/NIAID NIH HHS/United States ; R01 AI091595/AI/NIAID NIH HHS/United States ; R01 AI076342/AI/NIAID NIH HHS/United States ; R01 AI050529/AI/NIAID NIH HHS/United States ; R01 AI097137/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA Primers ; Drosophila melanogaster/microbiology ; Genetics, Population/*methods ; Genome, Bacterial ; Genomics/methods ; Humans ; Mycobacterium tuberculosis/genetics ; Sequence Analysis, DNA/*methods ; *Software ; Wolbachia/genetics ; }, abstract = {MOTIVATION: Population genomic analyses are often hindered by difficulties in obtaining sufficient numbers of genomes for analysis by DNA sequencing. Selective whole-genome amplification (SWGA) provides an efficient approach to amplify microbial genomes from complex backgrounds for sequence acquisition. However, the process of designing sets of primers for this method has many degrees of freedom and would benefit from an automated process to evaluate the vast number of potential primer sets.

RESULTS: Here, we present swga , a program that identifies primer sets for SWGA and evaluates them for efficiency and selectivity. We used swga to design and test primer sets for the selective amplification of Wolbachia pipientis genomic DNA from infected Drosophila melanogaster and Mycobacterium tuberculosis from human blood. We identify primer sets that successfully amplify each against their backgrounds and describe a general method for using swga for arbitrary targets. In addition, we describe characteristics of primer sets that correlate with successful amplification, and present guidelines for implementation of SWGA to detect new targets.

Source code and documentation are freely available on https://www.github.com/eclarke/swga . The program is implemented in Python and C and licensed under the GNU Public License.

CONTACT: ecl@mail.med.upenn.edu.

SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.}, } @article {pmid28332822, year = {2017}, author = {Khoshmanesh, A and Christensen, D and Perez-Guaita, D and Iturbe-Ormaetxe, I and O'Neill, SL and McNaughton, D and Wood, BR}, title = {Screening of Wolbachia Endosymbiont Infection in Aedes aegypti Mosquitoes Using Attenuated Total Reflection Mid-Infrared Spectroscopy.}, journal = {Analytical chemistry}, volume = {89}, number = {10}, pages = {5285-5293}, doi = {10.1021/acs.analchem.6b04827}, pmid = {28332822}, issn = {1520-6882}, mesh = {Aedes/chemistry/*microbiology ; Aging ; Animals ; Discriminant Analysis ; Female ; Least-Squares Analysis ; Male ; Sex Factors ; Spectroscopy, Fourier Transform Infrared/*methods ; Symbiosis ; Wolbachia/*pathogenicity/physiology ; }, abstract = {Dengue fever is the most common mosquito transmitted viral infection afflicting humans, estimated to generate around 390 million infections each year in over 100 countries. The introduction of the endosymbiotic bacterium Wolbachia into Aedes aegypti mosquitoes has the potential to greatly reduce the public health burden of the disease. This approach requires extensive polymerase chain reaction (PCR) testing of the Wolbachia-infection status of mosquitoes in areas where Wolbachia-A. aegypti are released. Here, we report the first example of small organism mid-infrared spectroscopy where we have applied attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectroscopy and multivariate modeling methods to determine sex, age, and the presence of Wolbachia (wMel strain) in laboratory mosquitoes and sex and age in field mosquitoes. The prediction errors using partial least squares discriminant analysis (PLS-DA) discrimination models for laboratory studies on independent test sets ranged from 0 to 3% for age and sex grading and 3% to 5% for Wolbachia infection diagnosis using dry mosquito abdomens while field study results using an artificial neural network yielded a 10% error. The application of FT-IR analysis is inexpensive, easy to use, and portable and shows significant potential to replace the reliance on more expensive and laborious PCR assays.}, } @article {pmid28303186, year = {2017}, author = {Sazama, EJ and Bosch, MJ and Shouldis, CS and Ouellette, SP and Wesner, JS}, title = {Incidence of Wolbachia in aquatic insects.}, journal = {Ecology and evolution}, volume = {7}, number = {4}, pages = {1165-1169}, pmid = {28303186}, issn = {2045-7758}, abstract = {Wolbachia is a genus of intracellular bacteria typically found within the reproductive systems of insects that manipulates those systems of their hosts. While current estimates of Wolbachia incidence suggest that it infects approximately half of all arthropod species, these estimates are based almost entirely on terrestrial insects. No systematic survey of Wolbachia in aquatic insects has been performed. To estimate Wolbachia incidence among aquatic insect species, we combined field-collected samples from the Missouri River (251 samples from 58 species) with a global database from previously published surveys. The final database contained 5,598 samples of 2,687 total species (228 aquatic and 2,459 terrestrial). We estimate that 52% (95% CrIs: 44%-60%) of aquatic insect species carry Wolbachia, compared to 60% (58%-63%) of terrestrial insects. Among aquatic insects, infected orders included Odonata, Coleoptera, Trichoptera, Ephemeroptera, Diptera, Hemiptera, and Plecoptera. Incidence was highest within aquatic Diptera and Hemiptera (69%), Odonata (50%), and Coleoptera (53%), and was lowest within Ephemeroptera (13%). These results indicate that Wolbachia is common among aquatic insects, but incidence varies widely across orders and is especially uncertain in those orders with low sample sizes such as Ephemeroptera, Plecoptera, and Trichoptera.}, } @article {pmid28303006, year = {2017}, author = {Aljayyoussi, G and Tyrer, HE and Ford, L and Sjoberg, H and Pionnier, N and Waterhouse, D and Davies, J and Gamble, J and Metuge, H and Cook, DAN and Steven, A and Sharma, R and Guimaraes, AF and Clare, RH and Cassidy, A and Johnston, KL and Myhill, L and Hayward, L and Wanji, S and Turner, JD and Taylor, MJ and Ward, SA}, title = {Short-Course, High-Dose Rifampicin Achieves Wolbachia Depletion Predictive of Curative Outcomes in Preclinical Models of Lymphatic Filariasis and Onchocerciasis.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {210}, pmid = {28303006}, issn = {2045-2322}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Administration, Oral ; Animals ; Anti-Bacterial Agents/*administration & dosage/pharmacology ; Brugia malayi/drug effects/microbiology/physiology ; DNA, Bacterial/drug effects ; Disease Models, Animal ; Elephantiasis, Filarial/*drug therapy/parasitology ; Embryonic Development/drug effects ; Filarioidea/drug effects/*microbiology/physiology ; Humans ; Mice ; Onchocerca volvulus/drug effects/microbiology/physiology ; Onchocerciasis/*drug therapy/parasitology ; Rifampin/*administration & dosage/pharmacology ; Treatment Outcome ; Wolbachia/*drug effects/genetics ; Wuchereria bancrofti/drug effects/microbiology/physiology ; }, abstract = {Lymphatic filariasis (LF) and onchocerciasis are priority neglected tropical diseases targeted for elimination. The only safe drug treatment with substantial curative activity against the filarial nematodes responsible for LF (Brugia malayi, Wuchereria bancrofti) or onchocerciasis (Onchocerca volvulus) is doxycycline. The target of doxycycline is the essential endosymbiont, Wolbachia. Four to six weeks doxycycline therapy achieves >90% depletion of Wolbachia in worm tissues leading to blockade of embryogenesis, adult sterility and premature death 18-24 months post-treatment. Long treatment length and contraindications in children and pregnancy are obstacles to implementing doxycycline as a public health strategy. Here we determine, via preclinical infection models of Brugia malayi or Onchocerca ochengi that elevated exposures of orally-administered rifampicin can lead to Wolbachia depletions from filariae more rapidly than those achieved by doxycycline. Dose escalation of rifampicin achieves >90% Wolbachia depletion in time periods of 7 days in B. malayi and 14 days in O. ochengi. Using pharmacokinetic-pharmacodynamic modelling and mouse-human bridging analysis, we conclude that clinically relevant dose elevations of rifampicin, which have recently been determined as safe in humans, could be administered as short courses to filariasis target populations with potential to reduce anti-Wolbachia curative therapy times to between one and two weeks.}, } @article {pmid28299861, year = {2017}, author = {Ma, WJ and Schwander, T}, title = {Patterns and mechanisms in instances of endosymbiont-induced parthenogenesis.}, journal = {Journal of evolutionary biology}, volume = {30}, number = {5}, pages = {868-888}, doi = {10.1111/jeb.13069}, pmid = {28299861}, issn = {1420-9101}, mesh = {Animals ; Diploidy ; Female ; Hymenoptera ; *Parthenogenesis ; *Symbiosis ; *Wolbachia/genetics/physiology ; }, abstract = {Female-producing parthenogenesis can be induced by endosymbionts that increase their transmission by manipulating host reproduction. Our literature survey indicates that such endosymbiont-induced parthenogenesis is known or suspected in 124 host species from seven different arthropod taxa, with Wolbachia as the most frequent endosymbiont (in 56-75% of host species). Most host species (81%, 100 out of 124) are characterized by haplo-diploid sex determination, but a strong ascertainment bias likely underestimates the frequency of endosymbiont-induced parthenogenesis in hosts with other sex determination systems. In at least one taxon, hymenopterans, endosymbionts are a significant driver of transitions from sexual to parthenogenetic reproduction, with one-third of lineages being parthenogenetic as a consequence of endosymbiont infection. Endosymbiont-induced parthenogenesis appears to facilitate the maintenance of reproductive polymorphism: at least 50% of species comprise both sexual (uninfected) and parthenogenetic (infected) strains. These strains feature distribution differences similar to the ones documented for lineages with genetically determined parthenogenesis, with endosymbiont-induced parthenogens occurring at higher latitudes than their sexual relatives. Finally, although gamete duplication is often considered as the main mechanism for endosymbiont-induced parthenogenesis, it underlies parthenogenesis in only half of the host species studied thus far. We point out caveats in the methods used to test for endosymbiont-induced parthenogenesis and suggest specific approaches that allow for firm conclusions about the involvement of endosymbionts in the origin of parthenogenesis.}, } @article {pmid28291780, year = {2017}, author = {Luck, AN and Slatko, BE and Foster, JM}, title = {Removing the needle from the haystack: Enrichment of Wolbachia endosymbiont transcripts from host nematode RNA by Cappable-seq™.}, journal = {PloS one}, volume = {12}, number = {3}, pages = {e0173186}, pmid = {28291780}, issn = {1932-6203}, mesh = {Animals ; Nematoda/*genetics/*microbiology ; RNA, Messenger/genetics ; RNA, Transfer/genetics ; Sequence Analysis, RNA/*methods ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Efficient transcriptomic sequencing of microbial mRNA derived from host-microbe associations is often compromised by the much lower relative abundance of microbial RNA in the mixed total RNA sample. One solution to this problem is to perform extensive sequencing until an acceptable level of transcriptome coverage is obtained. More cost-effective methods include use of prokaryotic and/or eukaryotic rRNA depletion strategies, sometimes in conjunction with depletion of polyadenylated eukaryotic mRNA. Here, we report use of Cappable-seq™ to specifically enrich, in a single step, Wolbachia endobacterial mRNA transcripts from total RNA prepared from the parasitic filarial nematode, Brugia malayi. The obligate Wolbachia endosymbiont is a proven drug target for many human filarial infections, yet the precise nature of its symbiosis with the nematode host is poorly understood. Insightful analysis of the expression levels of Wolbachia genes predicted to underpin the mutualistic association and of known drug target genes at different life cycle stages or in response to drug treatments is typically challenged by low transcriptomic coverage. Cappable-seq resulted in up to ~ 5-fold increase in the number of reads mapping to Wolbachia. On average, coverage of Wolbachia transcripts from B. malayi microfilariae was enriched ~40-fold by Cappable-seq. Additionally, this method has an additional benefit of selectively removing abundant prokaryotic ribosomal RNAs.The deeper microbial transcriptome sequencing afforded by Cappable-seq facilitates more detailed characterization of gene expression levels of pathogens and symbionts present in animal tissues.}, } @article {pmid28288761, year = {2017}, author = {Serrano-Parreño, B and Carretón, E and Caro-Vadillo, A and Falcón-Cordón, S and Falcón-Cordón, Y and Montoya-Alonso, JA}, title = {Pulmonary hypertension in dogs with heartworm before and after the adulticide protocol recommended by the American Heartworm Society.}, journal = {Veterinary parasitology}, volume = {236}, number = {}, pages = {34-37}, doi = {10.1016/j.vetpar.2017.02.001}, pmid = {28288761}, issn = {1873-2550}, mesh = {Animals ; Antinematodal Agents/*administration & dosage ; Arsenicals/administration & dosage ; Dirofilaria immitis/physiology ; Dirofilariasis/drug therapy/*parasitology/physiopathology ; Dog Diseases/drug therapy/parasitology/*physiopathology ; Dogs ; Doxycycline/administration & dosage ; Echocardiography/veterinary ; Endarteritis/drug therapy/parasitology/physiopathology/*veterinary ; Female ; Hypertension, Pulmonary/drug therapy/parasitology/physiopathology/*veterinary ; Injections, Intramuscular/veterinary ; Ivermectin/administration & dosage ; Male ; Spain ; Triazines/administration & dosage ; }, abstract = {Pulmonary hypertension (pH) is a frequent and severe phenomenon in heartworm disease (Dirofilaria immitis). There is a lack of studies assessing the evolution of the proliferative endarteritis and pH caused by D. immitis after the death of the parasites, so this study evaluated the influence that the elimination of the worms exerts over the pulmonary pressure and therefore evolution of the endarteritis, through the evaluation of the Right Pulmonary Artery Distensibility (RPAD) Index and other echocardiographic measurements in 2D mode, M-mode and Doppler echocardiography in 34 dogs naturally infected by D. immitis on day 0, and one month after the last adulticide dose (day 120). pH, based on the determination of the RPAD Index, was present in 68% of the dogs (n=23) on day 0 and on day 120. No significant differences were observed between the RPAD Index between the two measurements, and only significant differences were found in pulmonary deceleration time, ejection time, and left ventricular internal diameter in telediastole when measurements from day 0 and day 120 were compared. There was not any worsening in the development of pH after the elimination of the parasites, independently of the parasite burden. During the adulticide treatment, the death of the worms causes thromboembolism and tends to worsen the vascular damage and presence of pH . It seems that following the adulticide protocol recommended by the American Heartworm Society with the previous elimination of Wolbachia and reduction of microfilariae followed by the stepped death of the worms did not cause a significant aggravation of the pulmonary damage of the treated dogs. Neither is present any significant improvement in the RPAD Index on day 120; probably, more time is needed before appreciating some positive changes after the elimination of the worms and Wolbachia from the vasculature and further studies are necessary.}, } @article {pmid28286698, year = {2017}, author = {Yakob, L and Funk, S and Camacho, A and Brady, O and Edmunds, WJ}, title = {Aedes aegypti Control Through Modernized, Integrated Vector Management.}, journal = {PLoS currents}, volume = {9}, number = {}, pages = {}, pmid = {28286698}, issn = {2157-3999}, support = {206471/Z/17/Z/WT_/Wellcome Trust/United Kingdom ; MC_PC_15097/MRC_/Medical Research Council/United Kingdom ; MR/J01432X/1/MRC_/Medical Research Council/United Kingdom ; MR/K021680/1/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; }, abstract = {INTRODUCTION: In the context of the ongoing, unprecedented Zika virus outbreak in the Americas, the World Health Organization has expressed its support for developing and up-scaling three novel approaches to controlling the Aedes aegypti mosquito: the Sterile Insect Technique (SIT), the Release of Insects carrying Dominant Lethal genes (RIDL) and the release of Wolbachia-infected mosquitoes. Whereas the former two approaches are temporary insect population suppression strategies, Wolbachia infection is a self-sustaining, invasive strategy that uses inherited endosymbiotic bacteria to render natural mosquito populations arbovirus resistant.

METHODS: A mathematical model is parameterised with new, Brazilian field data informing the mating competitiveness of mass-reared, released insects; and simulations compare and contrast projections of vector control achieved with the alternative approaches.

RESULTS: Important disadvantages of Wolbachia and SIT are identified: both strategies result in mosquitoes ovipositing non-viable eggs and, by alleviating intense larval competition, can cause an overall increase in survival to the adult stage. However, it is demonstrated that strategically combining the suppression methods with Wolbachia can generate a sustained control while mitigating the risks of inadvertent exacerbation of the wild mosquito population.

DISCUSSION: This initial analysis demonstrates potential for good synergy when combining novel mosquito approaches in a modernized, integrated vector control programme.}, } @article {pmid28286343, year = {2017}, author = {Hofer, U}, title = {Symbiosis: Wolbachia's matchmaking secret revealed.}, journal = {Nature reviews. Microbiology}, volume = {15}, number = {4}, pages = {194-195}, pmid = {28286343}, issn = {1740-1534}, mesh = {Humans ; *Symbiosis ; *Wolbachia ; }, } @article {pmid28267749, year = {2017}, author = {Audsley, MD and Ye, YH and McGraw, EA}, title = {The microbiome composition of Aedes aegypti is not critical for Wolbachia-mediated inhibition of dengue virus.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {3}, pages = {e0005426}, pmid = {28267749}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*virology ; Animals ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Dengue Virus/*growth & development ; *Gastrointestinal Microbiome ; *Microbial Interactions ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/*growth & development ; }, abstract = {BACKGROUND: Dengue virus (DENV) is primarily vectored by the mosquito Aedes aegypti, and is estimated to cause 390 million human infections annually. A novel method for DENV control involves stable transinfection of Ae. aegypti with the common insect endosymbiont Wolbachia, which mediates an antiviral effect. However, the mechanism by which Wolbachia reduces the susceptibility of Ae. aegypti to DENV is not fully understood. In this study we assessed the potential of resident microbiota, which can play important roles in insect physiology and immune responses, to affect Wolbachia-mediated DENV blocking.

METHODOLOGY/FINDINGS: The microbiome of Ae. aegypti stably infected with Wolbachia strain wMel was compared to that of Ae. aegypti without Wolbachia, using 16s rDNA profiling. Our results indicate that although Wolbachia affected the relative abundance of several genera, the microbiome of both the Wolbachia-infected and uninfected mosquitoes was dominated by Elizabethkingia and unclassified Enterobacteriaceae. To assess the potential of the resident microbiota to affect the Wolbachia-mediated antiviral effect, we used antibiotic treatment before infection with DENV by blood-meal. In spite of a significant shift in the microbiome composition in response to the antibiotics, we detected no effect of antibiotic treatment on DENV infection rates, or on the DENV load of infected mosquitoes.

CONCLUSIONS/SIGNIFICANCE: Our findings indicate that stable infection with Wolbachia strain wMel produces few effects on the microbiome of laboratory-reared Ae. aegypti. Moreover, our findings suggest that the microbiome can be significantly altered without affecting the fundamental DENV blocking phenotype in these mosquitoes. Since Ae. aegypti are likely to encounter diverse microbiota in the field, this is a particularly important result in the context of using Wolbachia as a method for DENV control.}, } @article {pmid28266601, year = {2017}, author = {Wang, XX and Qi, LD and Jiang, R and Du, YZ and Li, YX}, title = {Incomplete removal of Wolbachia with tetracycline has two-edged reproductive effects in the thelytokous wasp Encarsia formosa (Hymenoptera: Aphelinidae).}, journal = {Scientific reports}, volume = {7}, number = {}, pages = {44014}, pmid = {28266601}, issn = {2045-2322}, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage ; Female ; Fertility/drug effects ; Gene Expression ; Male ; Oocytes/drug effects ; Reproduction ; Tetracycline/*administration & dosage ; Vitellogenins/genetics ; Wasps/*drug effects/*physiology ; Wolbachia/*drug effects/isolation & purification/pathogenicity ; }, abstract = {Wolbachia pipientis are intracellular endosymbionts that induce parthenogenesis in the parasitoid Encarsia formosa. Previous studies that focused on effects of Wolbachia on the wasp usually used tetracycline to remove Wolbachia without concern for the joint influences of tetracycline and Wolbachia. Here we treated the wasps (F0 lines) with tetracycline to produce offspring (F1 lines) which were not fed tetracycline to avoid antibiotic influence. The quantitative data and fluorescence in situ hybridization showed that Wolbachia titers were reduced but not totally removed. The Wolbachia that infected the male offspring were unpredictably detected. Low dose tetracycline enhanced the fertility of 2-day-old F0 wasps after 24 h of treatment; however, compared with controls, the oocyte load of 3- to 6-day-old tetracycline-treated wasps decreased day by day, and tetracycline reduced the longevity of the wasps. The fecundity of controls was significantly higher than that of the treated F1-10 and F1-20 generations. Gene expression of vitellogenin reflected the same trend as that of wasp fecundities in both F0 and F1 lines. Moreover, female offspring proportions of F0 and F1 lines were related to the titer of infected Wolbachia, demonstrating that Wolbachia titer affected the sex determination of E. formosa.}, } @article {pmid28262718, year = {2017}, author = {Terradas, G and Joubert, DA and McGraw, EA}, title = {The RNAi pathway plays a small part in Wolbachia-mediated blocking of dengue virus in mosquito cells.}, journal = {Scientific reports}, volume = {7}, number = {}, pages = {43847}, pmid = {28262718}, issn = {2045-2322}, mesh = {Aedes/immunology/microbiology/virology ; Animals ; Antibiosis/genetics/immunology ; Cell Line ; Dengue Virus/genetics/*immunology/physiology ; Gene Expression/immunology ; Host-Pathogen Interactions/genetics/immunology ; Mosquito Vectors/*immunology/microbiology/virology ; RNA Interference/*immunology ; Virus Replication/genetics/immunology ; Wolbachia/genetics/*immunology/physiology ; }, abstract = {Wolbachia pipientis is an insect endosymbiont known to limit the replication of viruses including dengue and Zika in their primary mosquito vector, Aedes aegypti. Wolbachia is being released into mosquito populations globally in a bid to control the diseases caused by these viruses. It is theorized that Wolbachia's priming of the insect immune system may confer protection against subsequent viral infection. Other hypotheses posit a role for competition between Wolbachia and viruses for host cellular resources. Using an A. aegypti cell line infected with Wolbachia, we tested the effects of targeting siRNAs against the major innate immune pathways on dengue virus loads. We show that while Wolbachia infection induces genes in the Toll, JAK/STAT and RNAi pathways, only reduced expression of RNAi leads to a rebound of dengue virus loads in Wolbachia-infected cells. The magnitude of the effect explained less than 10% of the total DENV load, demonstrating that blocking must be dependent on other factors in addition to the expression of RNAi. The findings bode well for the long-term stability of blocking given that immunity gene expression would likely be highly plastic and susceptible to rapid evolution.}, } @article {pmid28261755, year = {2017}, author = {Ramalho, MO and Martins, C and Silva, LM and Martins, VG and Bueno, OC}, title = {Intracellular Symbiotic Bacteria of Camponotus textor, Forel (Hymenoptera, Formicidae).}, journal = {Current microbiology}, volume = {74}, number = {5}, pages = {589-597}, pmid = {28261755}, issn = {1432-0991}, mesh = {Animals ; Ants/*microbiology ; *Bacteria/classification/genetics ; *Bacterial Physiological Phenomena ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; Wolbachia/classification/genetics ; }, abstract = {This study focuses on the weaver ant, Camponotus textor, Forel which occurs in some areas of the Brazilian Cerrado and Atlantic Forest, and its symbionts: Blochmannia, an obligate symbiont of Camponotus, and Wolbachia, known for causing reproductive alterations in their hosts. The main goal of this study was to investigate the presence, frequency of occurrence, and diversity of Wolbachia and Blochmannia strains in C. textor colonies. We found high infection rates (100%) and the occurrence of at least two distinct strains of Blochmannia (H_1 or H_7) in the same species. The observed haplotype variation within a single species may result from the high mutation rate of the symbiont. Similarly, the Wolbachia was found in all colonies with different rates of infections and a new strain (supergroup A) was deposited in the MLST database. The diversity found in the present study shows that there is still much to explore to understand about these symbiotic interactions.}, } @article {pmid28248294, year = {2017}, author = {Beckmann, JF and Ronau, JA and Hochstrasser, M}, title = {A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility.}, journal = {Nature microbiology}, volume = {2}, number = {}, pages = {17007}, pmid = {28248294}, issn = {2058-5276}, support = {R01 GM053756/GM/NIGMS NIH HHS/United States ; UL1 TR001863/TR/NCATS NIH HHS/United States ; }, mesh = {Animals ; Culex/*microbiology/*physiology ; Deubiquitinating Enzymes/*metabolism ; Drosophila/*microbiology/*physiology ; Male ; Reproduction ; Spermatozoa/microbiology/physiology ; Wolbachia/*enzymology/growth & development/metabolism ; }, abstract = {Wolbachia are obligate intracellular bacteria[1] that infect arthropods, including approximately two-thirds of insect species[2]. Wolbachia manipulate insect reproduction by enhancing their inheritance through the female germline. The most common alteration is cytoplasmic incompatibility (CI)[3-5], where eggs from uninfected females fail to develop when fertilized by sperm from Wolbachia-infected males. By contrast, if female and male partners are both infected, embryos are viable. CI is a gene-drive mechanism impacting population structure[6] and causing reproductive isolation[7], but its molecular mechanism has remained unknown. We show that a Wolbachia deubiquitylating enzyme (DUB) induces CI. The CI-inducing DUB, CidB, cleaves ubiquitin from substrates and is encoded in a two-gene operon, and the other protein, CidA, binds CidB. Binding is strongest between cognate partners in cidA-cidB homologues. In transgenic Drosophila, the cidA-cidB operon mimics CI when sperm introduce it into eggs, and a catalytically inactive DUB does not induce sterility. Toxicity is recapitulated in yeast by CidB alone; this requires DUB activity but is rescued by coexpressed CidA. A paralogous operon involves a putative nuclease (CinB) rather than a DUB. Analogous binding, toxicity and rescue in yeast were observed. These results identify a CI mechanism involving interacting proteins that are secreted into germline cells by Wolbachia, and suggest new methods for insect control.}, } @article {pmid28245239, year = {2017}, author = {Muturi, EJ and Ramirez, JL and Rooney, AP and Kim, CH}, title = {Comparative analysis of gut microbiota of mosquito communities in central Illinois.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {2}, pages = {e0005377}, pmid = {28245239}, issn = {1935-2735}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Culicidae/classification/*microbiology/physiology ; *Gastrointestinal Microbiome ; Gastrointestinal Tract/microbiology ; Illinois ; Insect Vectors/classification/*microbiology/physiology ; }, abstract = {BACKGROUND: The composition and structure of microbial communities that inhabit the mosquito midguts are poorly understood despite their well-documented potential to impede pathogen transmission.

We used MiSeq sequencing of the 16S rRNA gene to characterize the bacterial communities of field-collected populations of 12 mosquito species. After quality filtering and rarefaction, the remaining sequences were assigned to 181 operational taxonomic units (OTUs). Approximately 58% of these OTUs occurred in at least two mosquito species but only three OTUs: Gluconobacter (OTU 1), Propionibacterium (OTU 9), and Staphylococcus (OTU 31) occurred in all 12 mosquito species. Individuals of different mosquito species shared similar gut microbiota and it was common for individuals of the same species from the same study site and collection date to harbor different gut microbiota. On average, the microbiota of Aedes albopictus was the least diverse and significantly less even compared to Anopheles crucians, An. quadrimaculatus, Ae. triseriatus, Ae. vexans, Ae. japonicus, Culex restuans, and Culiseta inornata. The microbial community of Cx. pipiens and Ae. albopictus differed significantly from all other mosquitoes species and was primarily driven by the dominance of Wolbachia.

CONCLUSION AND SIGNIFICANCE: These findings expand the range of mosquito species whose gut microbiota has been characterized and sets the foundation for further studies to determine the influence of these microbiota on vector susceptibility to pathogens.}, } @article {pmid28241782, year = {2017}, author = {Vivero, RJ and Cadavid-Restrepo, G and Herrera, CX and Soto, SI}, title = {Molecular detection and identification of Wolbachia in three species of the genus Lutzomyia on the Colombian Caribbean coast.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {110}, pmid = {28241782}, issn = {1756-3305}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Caribbean Region ; Colombia ; Genotype ; Insect Vectors/microbiology ; Leishmaniasis/transmission ; Polymerase Chain Reaction ; Psychodidae/classification/genetics/*microbiology ; Wolbachia/*genetics/*isolation & purification ; }, abstract = {BACKGROUND: The hematophagous habits of insects belonging to the genus Lutzomyia (Diptera: Psychodidae), as well as their role as biological vectors of Leishmania species, make their presence an indication of infection risk. In the present study, seven species of Lutzomyia were identified and screened for natural infections with Wolbachia.

METHODS: Collection of sand flies was done in an endemic focus of leishmaniasis on the Colombian Caribbean coast (Department of Sucre, Ovejas municipality). DNA collected from Lutzomyia species was evaluated with PCR for wsp gene amplification to screen for bacterial infection.

RESULTS: Endosymbiotic Wolbachia was found in three species: Lutzomyia c. cayennensis, Lutzomyia dubitans and Lutzomyia evansi. Two Wolbachia strains (genotypes) were found in Lutzomyia spp. These genotypes were previously unknown in dipteran insects. The wLev strain was found in Lutzomyia dubitans, L. c. cayennensis and L. evansi and the wLcy strain was found only in L. c. cayennensis.

CONCLUSIONS: Genetic analysis indicated that the Wolbachia strains wLcy and wLev belong to the B Supergroup. This study provides evidence of infections of more than one strain of Wolbachia in L. c. cayennensis.}, } @article {pmid28241146, year = {2017}, author = {LePage, DP and Metcalf, JA and Bordenstein, SR and On, J and Perlmutter, JI and Shropshire, JD and Layton, EM and Funkhouser-Jones, LJ and Beckmann, JF and Bordenstein, SR}, title = {Prophage WO genes recapitulate and enhance Wolbachia-induced cytoplasmic incompatibility.}, journal = {Nature}, volume = {543}, number = {7644}, pages = {243-247}, pmid = {28241146}, issn = {1476-4687}, support = {R21 HD086833/HD/NICHD NIH HHS/United States ; P60 DK020593/DK/NIDDK NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; P30 EY008126/EY/NEI NIH HHS/United States ; U24 DK059637/DK/NIDDK NIH HHS/United States ; S10 OD021630/OD/NIH HHS/United States ; U2C DK059637/DK/NIDDK NIH HHS/United States ; T32 GM007347/GM/NIGMS NIH HHS/United States ; P30 DK020593/DK/NIDDK NIH HHS/United States ; P30 CA068485/CA/NCI NIH HHS/United States ; R01 AI081322/AI/NIAID NIH HHS/United States ; T32 GM008554/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified ; *Biological Control Agents ; Crosses, Genetic ; Cytoplasm/*genetics/pathology ; Drosophila melanogaster/*cytology/embryology/*microbiology/physiology ; Female ; Genes, Viral/*genetics ; *Host-Pathogen Interactions ; Male ; Prophages/*genetics ; Reproduction ; Sex Ratio ; Symbiosis ; Wolbachia/classification/*genetics/physiology/virology ; }, abstract = {The genus Wolbachia is an archetype of maternally inherited intracellular bacteria that infect the germline of numerous invertebrate species worldwide. They can selfishly alter arthropod sex ratios and reproductive strategies to increase the proportion of the infected matriline in the population. The most common reproductive manipulation is cytoplasmic incompatibility, which results in embryonic lethality in crosses between infected males and uninfected females. Females infected with the same Wolbachia strain rescue this lethality. Despite more than 40 years of research and relevance to symbiont-induced speciation, as well as control of arbovirus vectors and agricultural pests, the bacterial genes underlying cytoplasmic incompatibility remain unknown. Here we use comparative and transgenic approaches to demonstrate that two differentially transcribed, co-diverging genes in the eukaryotic association module of prophage WO from Wolbachia strain wMel recapitulate and enhance cytoplasmic incompatibility. Dual expression in transgenic, uninfected males of Drosophila melanogaster crossed to uninfected females causes embryonic lethality. Each gene additively augments embryonic lethality in crosses between infected males and uninfected females. Lethality associates with embryonic defects that parallel those of wild-type cytoplasmic incompatibility and is notably rescued by wMel-infected embryos in all cases. The discovery of cytoplasmic incompatibility factor genes cifA and cifB pioneers genetic studies of prophage WO-induced reproductive manipulations and informs the continuing use of Wolbachia to control dengue and Zika virus transmission to humans.}, } @article {pmid28235879, year = {2017}, author = {Erban, T and Ledvinka, O and Nesvorna, M and Hubert, J}, title = {Experimental Manipulation Shows a Greater Influence of Population than Dietary Perturbation on the Microbiome of Tyrophagus putrescentiae.}, journal = {Applied and environmental microbiology}, volume = {83}, number = {9}, pages = {}, pmid = {28235879}, issn = {1098-5336}, mesh = {Acaridae/*microbiology ; Animals ; Bacteria/*classification/*genetics ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Diet/*methods ; Feeding Behavior ; *Gastrointestinal Microbiome ; RNA, Ribosomal, 16S/genetics ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA ; }, abstract = {Tyrophagus putrescentiae is inhabited by bacteria that differ among mite populations (strains) and diets. Here, we investigated how the microbiome and fitness of Tputrescentiae are altered by dietary perturbations and mite populations. Four T. putrescentiae populations, referred to as dog, Koppert, laboratory, and Phillips, underwent a perturbation, i.e., a dietary switch from a rearing diet to two experimental diets. The microbiome was investigated by sequencing the V1-V3 portion of the 16S rRNA gene, and selected bacterial taxa were quantified by quantitative PCR (qPCR) using group/taxon-specific primers. The parameters observed were the changes in mite population growth and nutritional status, i.e., the total glycogen, lipid, saccharide, and protein contents in mites. The effect of diet perturbation on the variability of the microbiome composition and population growth was lower than the effect induced by mite population. In contrast, the diet perturbation showed a greater effect on nutritional status of mites than the mite population. The endosymbionts exhibited high variations among T. putrescentiae populations, including Cardinium in the laboratory population, Blattabacterium-like bacteria in the dog population, and Wolbachia in the dog and Phillips populations. Solitalea-like and Bartonella-like bacteria were present in the dog, Koppert, and Phillips populations in different proportions. The T. putrescentiae microbiome is dynamic and varies based on both the mite population and perturbation; however, the mites remain characterized by robust bacterial communities. Bacterial endosymbionts were found in all populations but represented a dominant portion of the microbiome in only some populations.IMPORTANCE We addressed the question of whether population origin or perturbation exerts a more significant influence on the bacterial community of the stored product mite Tyrophagus putrescentiae The microbiomes of four populations of T. putrescentiae insects subjected to diet perturbation were compared. Based on our results, the bacterial community was more affected by the mite population than by diet perturbation. This result can be interpreted as indicating high stability of the putative intracellular symbionts in response to dietary perturbation. The changes in the absolute and relative numbers of Wolbachia, Blattabacterium-like, Solitalea-like, and Cardinium bacteria in the T. putrescentiae populations can also be caused by neutral processes other than perturbation. When nutritional status is considered, the effect of population appeared less important than the perturbation. We hypothesize that differences in the proportions of the endosymbiotic bacteria result in changes in mite population growth.}, } @article {pmid28208639, year = {2017}, author = {Huang, YS and Higgs, S and Vanlandingham, DL}, title = {Biological Control Strategies for Mosquito Vectors of Arboviruses.}, journal = {Insects}, volume = {8}, number = {1}, pages = {}, pmid = {28208639}, issn = {2075-4450}, abstract = {Historically, biological control utilizes predatory species and pathogenic microorganisms to reduce the population of mosquitoes as disease vectors. This is particularly important for the control of mosquito-borne arboviruses, which normally do not have specific antiviral therapies available. Although development of resistance is likely, the advantages of biological control are that the resources used are typically biodegradable and ecologically friendly. Over the past decade, the advancement of molecular biology has enabled optimization by the manipulation of genetic materials associated with biological control agents. Two significant advancements are the discovery of cytoplasmic incompatibility induced by Wolbachia bacteria, which has enhanced replacement programs, and the introduction of dominant lethal genes into local mosquito populations through the release of genetically modified mosquitoes. As various arboviruses continue to be significant public health threats, biological control strategies have evolved to be more diverse and become critical tools to reduce the disease burden of arboviruses.}, } @article {pmid28204477, year = {2016}, author = {Lo, WS and Huang, YY and Kuo, CH}, title = {Winding paths to simplicity: genome evolution in facultative insect symbionts.}, journal = {FEMS microbiology reviews}, volume = {40}, number = {6}, pages = {855-874}, pmid = {28204477}, issn = {1574-6976}, mesh = {Animals ; Enterobacteriaceae/genetics ; *Evolution, Molecular ; Gene Transfer, Horizontal/genetics ; Genome, Bacterial/*genetics ; Insecta/*genetics/*microbiology ; Proteobacteria/genetics ; Spiroplasma/genetics ; Symbiosis/*genetics ; }, abstract = {Symbiosis between organisms is an important driving force in evolution. Among the diverse relationships described, extensive progress has been made in insect–bacteria symbiosis, which improved our understanding of the genome evolution in host-associated bacteria. Particularly, investigations on several obligate mutualists have pushed the limits of what we know about the minimal genomes for sustaining cellular life. To bridge the gap between those obligate symbionts with extremely reduced genomes and their non-host-restricted ancestors, this review focuses on the recent progress in genome characterization of facultative insect symbionts. Notable cases representing various types and stages of host associations, including those from multiple genera in the family Enterobacteriaceae (class Gammaproteobacteria), Wolbachia (Alphaproteobacteria) and Spiroplasma (Mollicutes), are discussed. Although several general patterns of genome reduction associated with the adoption of symbiotic relationships could be identified, extensive variation was found among these facultative symbionts. These findings are incorporated into the established conceptual frameworks to develop a more detailed evolutionary model for the discussion of possible trajectories. In summary, transitions from facultative to obligate symbiosis do not appear to be a universal one-way street; switches between hosts and lifestyles (e.g. commensalism, parasitism or mutualism) occur frequently and could be facilitated by horizontal gene transfer.}, } @article {pmid28201740, year = {2017}, author = {Bailly-Bechet, M and Martins-Simões, P and Szöllosi, GJ and Mialdea, G and Sagot, MF and Charlat, S}, title = {How Long Does Wolbachia Remain on Board?.}, journal = {Molecular biology and evolution}, volume = {34}, number = {5}, pages = {1183-1193}, doi = {10.1093/molbev/msx073}, pmid = {28201740}, issn = {1537-1719}, mesh = {Animals ; Arthropods/genetics ; DNA, Mitochondrial/genetics ; Evolution, Molecular ; Genetic Variation ; Genetics, Population ; Haplotypes ; Phylogeny ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Wolbachia bacteria infect about half of all arthropods, with diverse and extreme consequences ranging from sex-ratio distortion and mating incompatibilities to protection against viruses. These phenotypic effects, combined with efficient vertical transmission from mothers to offspring, satisfactorily explain the invasion dynamics of Wolbachia within species. However, beyond the species level, the lack of congruence between the host and symbiont phylogenetic trees indicates that Wolbachia horizontal transfers and extinctions do happen and underlie its global distribution. But how often do they occur? And has the Wolbachia pandemic reached its equilibrium? Here, we address these questions by inferring recent acquisition/loss events from the distribution of Wolbachia lineages across the mitochondrial DNA tree of 3,600 arthropod specimens, spanning 1,100 species from Tahiti and surrounding islands. We show that most events occurred within the last million years, but are likely attributable to individual level variation (e.g., imperfect maternal transmission) rather than population level variation (e.g., Wolbachia extinction). At the population level, we estimate that mitochondria typically accumulate 4.7% substitutions per site during an infected episode, and 7.1% substitutions per site during the uninfected phase. Using a Bayesian time calibration of the mitochondrial tree, these numbers translate into infected and uninfected phases of approximately 7 and 9 million years. Infected species thus lose Wolbachia slightly more often than uninfected species acquire it, supporting the view that its present incidence, estimated here slightly below 0.5, represents an epidemiological equilibrium.}, } @article {pmid28181227, year = {2017}, author = {Nguyen, DT and Morrow, JL and Spooner-Hart, RN and Riegler, M}, title = {Independent cytoplasmic incompatibility induced by Cardinium and Wolbachia maintains endosymbiont coinfections in haplodiploid thrips populations.}, journal = {Evolution; international journal of organic evolution}, volume = {71}, number = {4}, pages = {995-1008}, doi = {10.1111/evo.13197}, pmid = {28181227}, issn = {1558-5646}, mesh = {Animals ; Bacteroidetes/*physiology ; Reproduction ; *Symbiosis ; Thysanoptera/*microbiology ; Wolbachia/*physiology ; }, abstract = {Cardinium and Wolbachia are common maternally inherited reproductive parasites that can coinfect arthropods, yet interactions between both bacterial endosymbionts are rarely studied. For the first time, we report their independent expression of complete cytoplasmic incompatibility (CI) in a coinfected host, and CI in a species of the haplodiploid insect order Thysanoptera. In Pezothrips kellyanus, Cardinium-induced CI resulted in a combination of male development (MD) and embryonic female mortality (FM) of fertilized eggs. In contrast, Wolbachia-induced CI resulted in FM together with postembryonic mortality not previously reported as a CI outcome. Both endosymbionts appeared to not influence fecundity but virgins produced more offspring than mated females. In coinfected individuals, Wolbachia density was higher than Cardinium. Wolbachia removal did not impact Cardinium density, suggesting a lack of competition within hosts. Maternal transmission was complete for Wolbachia and high for Cardinium. Our data support theoretical predictions and empirical detection of high endosymbiont prevalence in field populations of the native range of this pest thrips. However, previous findings of more frequent loss of Wolbachia than Cardinium, particularly in field populations of the host's invasive range, suggest that genetic diversity or varying environmental factors between field populations also play a role in shaping host-endosymbiont dynamics.}, } @article {pmid28172869, year = {2016}, author = {Kraaijeveld, K and Anvar, SY and Frank, J and Schmitz, A and Bast, J and Wilbrandt, J and Petersen, M and Ziesmann, T and Niehuis, O and de Knijff, P and den Dunnen, JT and Ellers, J}, title = {Decay of Sexual Trait Genes in an Asexual Parasitoid Wasp.}, journal = {Genome biology and evolution}, volume = {8}, number = {12}, pages = {3685-3695}, pmid = {28172869}, issn = {1759-6653}, mesh = {Animals ; Female ; *Genes, Insect ; Male ; Mutation ; Phenotype ; Phylogeny ; Polymorphism, Single Nucleotide ; *Reproduction, Asexual ; Wasps/*genetics/physiology ; }, abstract = {Trait loss is a widespread phenomenon with pervasive consequences for a species’ evolutionary potential. The genetic changes underlying trait loss have only been clarified in a small number of cases. None of these studies can identify whether the loss of the trait under study was a result of neutral mutation accumulation or negative selection. This distinction is relatively clear-cut in the loss of sexual traits in asexual organisms. Male-specific sexual traits are not expressed and can only decay through neutral mutations, whereas female-specific traits are expressed and subject to negative selection. We present the genome of an asexual parasitoid wasp and compare it to that of a sexual lineage of the same species. We identify a short-list of 16 genes for which the asexual lineage carries deleterious SNP or indel variants, whereas the sexual lineage does not. Using tissue-specific expression data from other insects, we show that fifteen of these are expressed in male-specific reproductive tissues. Only one deleterious variant was found that is expressed in the female-specific spermathecae, a trait that is heavily degraded and thought to be under negative selection in L. clavipes. Although the phenotypic decay of male-specific sexual traits in asexuals is generally slow compared with the decay of female-specific sexual traits, we show that male-specific traits do indeed accumulate deleterious mutations as expected by theory. Our results provide an excellent starting point for detailed study of the genomics of neutral and selected trait decay.}, } @article {pmid28168034, year = {2017}, author = {Johannesen, J}, title = {Tracing the history and ecological context of Wolbachia double infection in a specialist host (Urophora cardui)-parasitoid (Eurytoma serratulae) system.}, journal = {Ecology and evolution}, volume = {7}, number = {3}, pages = {986-996}, pmid = {28168034}, issn = {2045-7758}, abstract = {The endosymbiotic bacterium Wolbachia is the most widespread bacteria in insects, yet the ecology of novel acquisitions in natural host populations is poorly understood. Using temporal data separated by 12 years, I tested the hypothesis that immigration of a parasitoid wasp led to transmission of its Wolbachia strain to its dipteran host, resulting in double-strain infection, and I used geographic and community surveys to explore the history of transmission in fly and parasitoid. Double infection in the fly host was present before immigration of the parasitoid. Equal prevalence of double infection in males and females, constant prevalence before and after immigration in two regions, and increase in one region of immigration indicate little if no competition between strains. Double infection was present throughout the fly's distribution range, but proportions varied highly (0-0.71, mean = 0.26). Two fly-specific MLST strains, observed in Eastern and Western Europe, respectively, differed at hcpA only. Flies with either fly-strain could be double infected with the parasitoid's strain. The geographic distribution of double infection implies that it is older than the fly host's extent distribution range and that different proportions of double infection are caused by demographic fluctuations in the fly. The geographic data in combination with community surveys of infections and strains further suggest that the parasitoid strain was the fly's ancestral strain that was transmitted to the parasitoid, that is, the reverse transmission route as first hypothesized. Based on these findings together with a comparison of oviposition strategies of other hosts harboring related Wolbachia strains, I hypothesize that trans-infection during an insect host's puparial metamorphosis might be important in promoting horizontal transmission among diverse holometabolic taxa.}, } @article {pmid28159754, year = {2017}, author = {White, PM and Serbus, LR and Debec, A and Codina, A and Bray, W and Guichet, A and Lokey, RS and Sullivan, W}, title = {Reliance of Wolbachia on High Rates of Host Proteolysis Revealed by a Genome-Wide RNAi Screen of Drosophila Cells.}, journal = {Genetics}, volume = {205}, number = {4}, pages = {1473-1488}, pmid = {28159754}, issn = {1943-2631}, support = {R01 GM104486/GM/NIGMS NIH HHS/United States ; T34 GM007910/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cell Line ; Drosophila/*genetics/metabolism/microbiology ; Drosophila Proteins/genetics/metabolism ; Endoplasmic Reticulum-Associated Degradation ; *Genome, Insect ; Host-Pathogen Interactions/*genetics ; Lipid Metabolism ; Mitochondria/metabolism ; *Proteolysis ; RNA Interference ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia are gram-negative, obligate, intracellular bacteria carried by a majority of insect species worldwide. Here we use a Wolbachia-infected Drosophila cell line and genome-wide RNA interference (RNAi) screening to identify host factors that influence Wolbachia titer. By screening an RNAi library targeting 15,699 transcribed host genes, we identified 36 candidate genes that dramatically reduced Wolbachia titer and 41 that increased Wolbachia titer. Host gene knockdowns that reduced Wolbachia titer spanned a broad array of biological pathways including genes that influenced mitochondrial function and lipid metabolism. In addition, knockdown of seven genes in the host ubiquitin and proteolysis pathways significantly reduced Wolbachia titer. To test the in vivo relevance of these results, we found that drug and mutant inhibition of proteolysis reduced levels of Wolbachia in the Drosophila oocyte. The presence of Wolbachia in either cell lines or oocytes dramatically alters the distribution and abundance of ubiquitinated proteins. Functional studies revealed that maintenance of Wolbachia titer relies on an intact host Endoplasmic Reticulum (ER)-associated protein degradation pathway (ERAD). Accordingly, electron microscopy studies demonstrated that Wolbachia is intimately associated with the host ER and dramatically alters the morphology of this organelle. Given Wolbachia lack essential amino acid biosynthetic pathways, the reliance of Wolbachia on high rates of host proteolysis via ubiquitination and the ERAD pathways may be a key mechanism for provisioning Wolbachia with amino acids. In addition, the reliance of Wolbachia on the ERAD pathway and disruption of ER morphology suggests a previously unsuspected mechanism for Wolbachia's potent ability to prevent RNA virus replication.}, } @article {pmid28156001, year = {2017}, author = {Joob, B and Wiwanitkit, V}, title = {Zika virus infection and Wolbachia (comment on DOI: 10.1002/bies.201600175).}, journal = {BioEssays : news and reviews in molecular, cellular and developmental biology}, volume = {39}, number = {4}, pages = {}, doi = {10.1002/bies.201600267}, pmid = {28156001}, issn = {1521-1878}, mesh = {Humans ; *Wolbachia ; Zika Virus ; *Zika Virus Infection ; }, } @article {pmid28152011, year = {2017}, author = {Tsujimoto, H and Hanley, KA and Sundararajan, A and Devitt, NP and Schilkey, FD and Hansen, IA}, title = {Dengue virus serotype 2 infection alters midgut and carcass gene expression in the Asian tiger mosquito, Aedes albopictus.}, journal = {PloS one}, volume = {12}, number = {2}, pages = {e0171345}, pmid = {28152011}, issn = {1932-6203}, support = {P20 GM103451/GM/NIGMS NIH HHS/United States ; R15 AI113628/AI/NIAID NIH HHS/United States ; SC1 AI109055/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/microbiology/*virology ; Animals ; *Dengue Virus ; Female ; Gastrointestinal Tract/metabolism/*virology ; Gene Expression Regulation, Viral ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Wolbachia ; }, abstract = {BACKGROUND: The Asian tiger mosquito, Aedes albopictus is currently an important vector for dengue, chikungunya and Zika virus, and its role in transmission of arthropod-borne viruses (arboviruses) may increase in the future due to its ability to colonize temperate regions. In contrast to Aedes aegypti, the dominant vector of dengue, chikungunya and Zika virus, genetic responses of Ae. albopictus upon infection with an arbovirus are not well characterized. Here we present a study of the changes in transcript expression in Ae. albopictus exposed to dengue virus serotype 2 via feeding on an artificial bloodmeal.

We isolated midguts and midgut-free carcasses of Ae. albopictus fed on bloodmeals containing dengue virus as well as controls fed on virus-free control meals at day 1 and day 5 post-feeding. We confirmed infection of midguts from mosquitoes sampled on day 5 post-feeding via RT-PCR. RNAseq analysis revealed dynamic modulation of the expression of several putative immunity and dengue virus-responsive genes, some of whose expression was verified by qRT-PCR. For example, a serine protease gene was up-regulated in the midgut at 1 day post infection, which may potentially enhance mosquito susceptibility to dengue infection, while 14 leucine-rich repeat genes, previously shown to be involved in mosquito antiviral defenses, were down-regulated in the carcass at 5 days post infection. The number of significantly modulated genes decreased over time in midguts and increased in carcasses.

CONCLUSION/SIGNIFICANCE: Dengue virus exposure results in the modulation of genes in a time- and site-specific manner. Previous literature on the interaction between mosquitoes and mosquito-borne pathogens suggests that most of the changes that occurred in Ae. albopictus exposed to DENV would favor virus infection. Many genes identified in this study warrant further characterization to understand their role in viral manipulation of and antiviral response of Ae. albopictus.}, } @article {pmid28135281, year = {2017}, author = {Medeiros, MC and Boothe, EC and Roark, EB and Hamer, GL}, title = {Dispersal of male and female Culex quinquefasciatus and Aedes albopictus mosquitoes using stable isotope enrichment.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {1}, pages = {e0005347}, pmid = {28135281}, issn = {1935-2735}, mesh = {Aedes/*physiology ; *Animal Distribution ; Animals ; Culex/*physiology ; Female ; Isotope Labeling ; Larva/physiology ; Male ; Mosquito Vectors/*physiology ; Oviposition/physiology ; Texas ; }, abstract = {The dispersal patterns of mosquito vectors are important drivers of vector-borne infectious disease dynamics and understanding movement patterns is pivotal to devise successful intervention strategies. Here, we investigate the dispersal patterns of two globally important mosquito vectors, Aedes albopictus and Culex quinquefasciatus, by marking naturally-occurring larvae with stable isotopes (13C or 15N). Marked individuals were captured with 32 CDC light trap, 32 gravid trap, and 16 BG Sentinel at different locations within two-kilometer radii of six larval habitats enriched with either 13C or 15N. In total, 720 trap nights from July to August 2013 yielded a total of 32,140 Cx. quinquefasciatus and 7,722 Ae. albopictus. Overall, 69 marked female mosquitoes and 24 marked male mosquitoes were captured throughout the study period. The distance that Cx. quinquefasciatus females traveled differed for host-seeking and oviposition-seeking traps, with females seeking oviposition sites traveling further than those seeking hosts. Our analysis suggests that 41% of Cx. quinquefasciatus females that were host-seeking occurred 1-2 kilometer from their respective natal site, while 59% remained within a kilometer of their natal site. In contrast, 59% of Cx. quinquefasciatus females that were seeking oviposition sites occurred between 1-2 kilometer away from their larval habitat, while 15% occurred > 2 kilometer away from their natal site. Our analysis estimated that approximately 100% of Ae. albopictus females remained within 1 km of their respective natal site, with 79% occurring within 250m. In addition, we found that male Ae. albopictus dispersed farther than females, suggesting male-biased dispersal in this Ae. albopictus population. This study provides important insights on the dispersal patterns of two globally relevant vector species, and will be important in planning next generation vector control strategies that mitigate mosquito-borne disease through sterile insect techniques, novel Wolbachia infection, and gene drive strategies.}, } @article {pmid28127053, year = {2017}, author = {Pekas, A and Palevsky, E and Sumner, JC and Perotti, MA and Nesvorna, M and Hubert, J}, title = {Comparison of bacterial microbiota of the predatory mite Neoseiulus cucumeris (Acari: Phytoseiidae) and its factitious prey Tyrophagus putrescentiae (Acari: Acaridae).}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {2}, pmid = {28127053}, issn = {2045-2322}, mesh = {Acari/*microbiology ; Animals ; Bacteria/*classification/*genetics ; Metagenomics ; *Microbiota ; Symbiosis ; }, abstract = {Neoseiulus cucumeris is a predatory mite used for biological control of arthropod pests. Mass-reared predators are fed with factitious prey mites such as Tyrophagus putrescentiae. Although some information on certain endosymbionts of N. cucumeris and T. putrescentiae exists, it is unclear whether both species share bacterial communities. The bacterial communities in populations of predator and prey mites, as well as the occurence of potential acaropathogenic bacteria were analyzed. The comparisons were based on the following groups: (i) N. cucumeris mass-production; (ii) N. cucumeris laboratory population with disease symptoms; (iii) T. putrescentiae pure populations and; (iv) T. putrescentiae from rearing units of N. cucumeris. Only 15% of OTUs were present in all samples from predatory and prey mite populations (core OTUs): the intracellular symbionts Wolbachia, Cardinium, plus other Blattabacterium-like, Solitalea-like, and Bartonella-like symbionts. Environmental bacteria were more abundant in predatory mites, while symbiotic bacteria prevailed in prey mites. Relative numbers of certain bacterial taxa were significantly different between the microbiota of prey mites reared with and without N. cucumeris. No significant differences were found in the bacterial communities of healthy N. cucumeris compared to N. cucumeris showing disease symptoms. We did not identify any confirmed acaropathogenic bacteria among microbiota.}, } @article {pmid28118850, year = {2017}, author = {Jha, R and Gangwar, M and Chahar, D and Setty Balakrishnan, A and Negi, MP and Misra-Bhattacharya, S}, title = {Humans from Wuchereria bancrofti endemic area elicit substantial immune response to proteins of the filarial parasite Brugia malayi and its endosymbiont Wolbachia.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {40}, pmid = {28118850}, issn = {1756-3305}, mesh = {Animals ; Antibodies, Bacterial/analysis/immunology ; Antibodies, Helminth/analysis/immunology ; Bacterial Proteins/analysis/*immunology ; Brugia malayi/genetics/*immunology/*microbiology ; Elephantiasis, Filarial/*immunology/parasitology ; Enzyme-Linked Immunosorbent Assay ; Female ; Filariasis/*immunology/parasitology ; Helminth Proteins/analysis/*immunology ; Humans ; Immunity, Humoral ; Immunoblotting ; Leukocytes, Mononuclear/immunology/parasitology ; Symbiosis ; Wolbachia/*immunology/physiology ; Wuchereria bancrofti/genetics/*immunology ; }, abstract = {BACKGROUND: In the past, immune responses to several Brugia malayi immunodominant antigens have been characterized in filaria-infected populations; however, little is known regarding Wolbachia proteins. We earlier cloned and characterized few B. malayi (trehalose-6-phosphate phosphatase, Bm-TPP and heavy chain myosin, BmAF-Myo) and Wolbachia (translation initiation factor-1, Wol Tl IF-1 and NAD[+]-dependent DNA ligase, wBm-LigA) proteins and investigated the immune responses, which they triggered in animal models. The current study emphasizes on immunological characteristics of these proteins in three major categories of filarial endemic zones: endemic normal (EN, asymptomatic, amicrofilaraemic; putatively immune), microfilariae carriers (MF, asymptomatic but microfilaraemic), and chronic filarial patients (CP, symptomatic and mostly amicrofilaraemic).

METHODS: Immunoblotting and ELISA were carried out to measure IgG and isotype antibodies against these recombinant proteins in various clinical categories. Involvement of serum antibodies in infective larvae killing was assessed by antibody-dependent cellular adhesion and cytotoxicity assay. Cellular immune response was investigated by in vitro proliferation of peripheral blood mononuclear cells (PBMCs) and reactive oxygen species (ROS) generation in these cells after stimulation.

RESULTS: Immune responses of EN and CP displayed almost similar level of IgG to Wol Tl IF-1 while other three proteins had higher serum IgG in EN individuals only. Specific IgA, IgG1, IgG3 and IgM to Bm-TPP were high in EN subjects, while BmAF-Myo additionally showed elevated IgG2. Enhanced IgA and IgG3 were detected in both EN and CP individuals in response to Wol Tl IF-1 antigen, but IgG1 and IgM were high only in EN individuals. wBm-LigA and BmAF-Myo exhibited almost similar pattern of antibody responses. PBMC isolated from EN subjects exhibited higher proliferation and ROS generation when stimulated with all three proteins except for Wol Tl IF-1.

CONCLUSIONS: Overall, these findings display high immunogenicity of all four proteins in human subjects and revealed that the EN population was exposed to both B. malayi and Wolbachia proteins simultaneously. In addition, immune responses to Wol Tl IF-1 suggest possible role of this factor in Wolbachia-induced pathological responses while immune responses to other three proteins suggest that these can be explored further as vaccine candidates.}, } @article {pmid28118586, year = {2017}, author = {Pontier, SM and Schweisguth, F}, title = {Response to "Does pupal communication influence Wolbachia-mediated cytoplasmic incompatibility?".}, journal = {Current biology : CB}, volume = {27}, number = {2}, pages = {R55-R56}, doi = {10.1016/j.cub.2016.12.013}, pmid = {28118586}, issn = {1879-0445}, mesh = {Animals ; Drosophila ; Drosophila melanogaster ; Female ; Germ Cells ; Male ; Pupa ; *Wolbachia ; }, abstract = {In a recent Current Biology paper [1], we reported that pheromone communication occurred during metamorphosis in Drosophila melanogaster. Female pheromones appeared to influence various aspects of the physiology and development of adult males. In particular, we observed that this communication regulated testis development and had a positive impact on reproduction, as measured by a difference in the % of eggs developing into larvae in crosses involving adult male flies that had developed at metamorphosis with or without female pupae [1].}, } @article {pmid28118585, year = {2017}, author = {Jacquet, A and Horard, B and Loppin, B}, title = {Does pupal communication influence Wolbachia-mediated cytoplasmic incompatibility?.}, journal = {Current biology : CB}, volume = {27}, number = {2}, pages = {R53-R55}, doi = {10.1016/j.cub.2016.12.012}, pmid = {28118585}, issn = {1879-0445}, mesh = {Animals ; Cytoplasm ; Drosophila ; Drosophila melanogaster ; Female ; Male ; Pupa ; *Wolbachia ; }, abstract = {Wolbachia are widespread endosymbiotic bacteria found in terrestrial arthropods and filarial nematodes [1]. In insects, Wolbachia generally rely on diverse strategies to manipulate their host's reproduction and favor their own vertical transmission through infected eggs [2]. One such mechanism is a sterility syndrome called 'cytoplasmic incompatibility'. Cytoplasmic incompatibility occurs at fertilization, when a spermatozoon from a Wolbachia-infected male fertilizes an uninfected egg. In this case, sperm-derived chromosomes fail to separate normally at the first zygotic division, thus preventing the development of a diploid embryo [3]. Moreover, the presence of Wolbachia in females rescues the integration of paternal chromosomes in the zygote and allows the development of a viable, infected individual. Although the molecular basis of cytoplasmic incompatibility is still unknown, a current model implies the existence of Wolbachia-induced reversible modifications on sperm DNA or chromatin that must be eliminated or neutralized shortly after fertilization by rescuing Wolbachia factors present in infected eggs [4]. In a recent Current Biology paper [5], Stéphanie Pontier and François Schweisguth recently challenged this model by proposing that Wolbachia perturbs a pheromone-based communication between male and female pupae in Drosophila melanogaster and Drosophila simulans, which controls the "compatibility range" of male and female gametes. However, we fail to detect any influence of pupal communication on cytoplasmic incompatibility in Drosophila simulans as well as in the parasitoid wasp Nasonia vitripennis. Our results thus question the robustness of their model.}, } @article {pmid28106771, year = {2017}, author = {Staudacher, H and Schimmel, BC and Lamers, MM and Wybouw, N and Groot, AT and Kant, MR}, title = {Independent Effects of a Herbivore's Bacterial Symbionts on Its Performance and Induced Plant Defences.}, journal = {International journal of molecular sciences}, volume = {18}, number = {1}, pages = {}, pmid = {28106771}, issn = {1422-0067}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Bacteria/*drug effects ; Female ; *Herbivory/drug effects ; Solanum lycopersicum/immunology/*microbiology/*parasitology ; Male ; Movement ; Plant Growth Regulators/analysis ; Survival Analysis ; *Symbiosis/drug effects ; Tetranychidae/drug effects/*physiology ; }, abstract = {It is well known that microbial pathogens and herbivores elicit defence responses in plants. Moreover, microorganisms associated with herbivores, such as bacteria or viruses, can modulate the plant's response to herbivores. Herbivorous spider mites can harbour different species of bacterial symbionts and exert a broad range of effects on host-plant defences. Hence, we tested the extent to which such symbionts affect the plant's defences induced by their mite host and assessed if this translates into changes in plant resistance. We assessed the bacterial communities of two strains of the common mite pest Tetranychus urticae. We found that these strains harboured distinct symbiotic bacteria and removed these using antibiotics. Subsequently, we tested to which extent mites with and without symbiotic bacteria induce plant defences in terms of phytohormone accumulation and defence gene expression, and assessed mite oviposition and survival as a measure for plant resistance. We observed that the absence/presence of these bacteria altered distinct plant defence parameters and affected mite performance but we did not find indications for a causal link between the two. We argue that although bacteria-related effects on host-induced plant defences may occur, these do not necessarily affect plant resistance concomitantly.}, } @article {pmid28100819, year = {2017}, author = {Longdon, B and Day, JP and Schulz, N and Leftwich, PT and de Jong, MA and Breuker, CJ and Gibbs, M and Obbard, DJ and Wilfert, L and Smith, SC and McGonigle, JE and Houslay, TM and Wright, LI and Livraghi, L and Evans, LC and Friend, LA and Chapman, T and Vontas, J and Kambouraki, N and Jiggins, FM}, title = {Vertically transmitted rhabdoviruses are found across three insect families and have dynamic interactions with their hosts.}, journal = {Proceedings. Biological sciences}, volume = {284}, number = {1847}, pages = {}, pmid = {28100819}, issn = {1471-2954}, support = {281668/ERC_/European Research Council/International ; BB/K000489/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 109356/Z/15/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Butterflies/virology ; Ceratitis capitata/virology ; Drosophila/virology ; *Infectious Disease Transmission, Vertical ; Insecta/*virology ; *Rhabdoviridae ; }, abstract = {A small number of free-living viruses have been found to be obligately vertically transmitted, but it remains uncertain how widespread vertically transmitted viruses are and how quickly they can spread through host populations. Recent metagenomic studies have found several insects to be infected with sigma viruses (Rhabdoviridae). Here, we report that sigma viruses that infect Mediterranean fruit flies (Ceratitis capitata), Drosophila immigrans, and speckled wood butterflies (Pararge aegeria) are all vertically transmitted. We find patterns of vertical transmission that are consistent with those seen in Drosophila sigma viruses, with high rates of maternal transmission, and lower rates of paternal transmission. This mode of transmission allows them to spread rapidly in populations, and using viral sequence data we found the viruses in D. immigrans and C. capitata had both recently swept through host populations. The viruses were common in nature, with mean prevalences of 12% in C. capitata, 38% in D. immigrans and 74% in P. aegeria We conclude that vertically transmitted rhabdoviruses may be widespread in a broad range of insect taxa, and that these viruses can have dynamic interactions with their hosts.}, } @article {pmid28097419, year = {2017}, author = {Farkas, JZ and Gourley, SA and Liu, R and Yakubu, AA}, title = {Modelling Wolbachia infection in a sex-structured mosquito population carrying West Nile virus.}, journal = {Journal of mathematical biology}, volume = {75}, number = {3}, pages = {621-647}, pmid = {28097419}, issn = {1432-1416}, mesh = {Aedes/*microbiology/*virology ; Animals ; Female ; Humans ; Insect Vectors/*microbiology/*virology ; Male ; Models, Biological ; Rickettsiaceae Infections/prevention & control/transmission ; Sex Factors ; West Nile Fever/prevention & control/transmission ; West Nile virus/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is possibly the most studied reproductive parasite of arthropod species. It appears to be a promising candidate for biocontrol of some mosquito borne diseases. We begin by developing a sex-structured model for a Wolbachia infected mosquito population. Our model incorporates the key effects of Wolbachia infection including cytoplasmic incompatibility and male killing. We also allow the possibility of reduced reproductive output, incomplete maternal transmission, and different mortality rates for uninfected/infected male/female individuals. We study the existence and local stability of equilibria, including the biologically relevant and interesting boundary equilibria. For some biologically relevant parameter regimes there may be multiple coexistence steady states including, very importantly, a coexistence steady state in which Wolbachia infected individuals dominate. We also extend the model to incorporate West Nile virus (WNv) dynamics, using an SEI modelling approach. Recent evidence suggests that a particular strain of Wolbachia infection significantly reduces WNv replication in Aedes aegypti. We model this via increased time spent in the WNv-exposed compartment for Wolbachia infected female mosquitoes. A basic reproduction number [Formula: see text] is computed for the WNv infection. Our results suggest that, if the mosquito population consists mainly of Wolbachia infected individuals, WNv eradication is likely if WNv replication in Wolbachia infected individuals is sufficiently reduced.}, } @article {pmid28087534, year = {2017}, author = {White, PM and Pietri, JE and Debec, A and Russell, S and Patel, B and Sullivan, W}, title = {Mechanisms of Horizontal Cell-to-Cell Transfer of Wolbachia spp. in Drosophila melanogaster.}, journal = {Applied and environmental microbiology}, volume = {83}, number = {7}, pages = {}, pmid = {28087534}, issn = {1098-5336}, support = {R01 GM104486/GM/NIGMS NIH HHS/United States ; R25 GM056765/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Clathrin/metabolism ; Drosophila melanogaster/*cytology/*microbiology/physiology ; Dynamins/metabolism ; Germ Cells/microbiology ; In Situ Hybridization, Fluorescence ; Wolbachia/cytology/*physiology ; }, abstract = {Wolbachia is an intracellular endosymbiont present in most arthropod and filarial nematode species. Transmission between hosts is primarily vertical, taking place exclusively through the female germ line, although horizontal transmission has also been documented. The results of several studies indicate that Wolbachia spp. can undergo transfer between somatic and germ line cells during nematode development and in adult flies. However, the mechanisms underlying horizontal cell-to-cell transfer remain largely unexplored. Here, we establish a tractable system for probing horizontal transfer of Wolbachia cells between Drosophila melanogaster cells in culture using fluorescence in situ hybridization (FISH). First, we show that horizontal transfer is independent of cell-to-cell contact and can efficiently take place through the culture medium within hours. Further, we demonstrate that efficient transfer utilizes host cell phagocytic and clathrin/dynamin-dependent endocytic machinery. Lastly, we provide evidence that this process is conserved between species, showing that horizontal transfer from mosquito to Drosophila cells takes place in a similar fashion. Altogether, our results indicate that Wolbachia utilizes host internalization machinery during infection, and this mechanism is conserved across insect species.IMPORTANCE Our work has broad implications for the control and treatment of tropical diseases. Wolbachia can confer resistance against a variety of human pathogens in mosquito vectors. Elucidating the mechanisms of horizontal transfer will be useful for efforts to more efficiently infect nonnatural insect hosts with Wolbachia as a biological control agent. Further, as Wolbachia is essential for the survival of filarial nematodes, understanding horizontal transfer might provide new approaches to treating human infections by targeting Wolbachia Finally, this work provides a key first step toward the genetic manipulation of Wolbachia.}, } @article {pmid28085049, year = {2017}, author = {Mariño, YA and Verle Rodrigues, JC and Bayman, P}, title = {Wolbachia Affects Reproduction and Population Dynamics of the Coffee Berry Borer (Hypothenemus hampei): Implications for Biological Control.}, journal = {Insects}, volume = {8}, number = {1}, pages = {}, pmid = {28085049}, issn = {2075-4450}, support = {P20 GM103475/GM/NIGMS NIH HHS/United States ; P20 RR016470/RR/NCRR NIH HHS/United States ; }, abstract = {Wolbachia are widely distributed endosymbiotic bacteria that influence the reproduction and fitness of their hosts. In recent years the manipulation of Wolbachia infection has been considered as a potential tool for biological control. The coffee berry borer (CBB), Hypothenemus hampei, is the most devastating coffee pest worldwide. Wolbachia infection in the CBB has been reported, but until now the role of Wolbachia in CBB reproduction and fitness has not been tested. To address this issue we reared the CBB in artificial diets with and without tetracycline (0.1% w/v) for ten generations. Tetracycline reduced significantly the relative proportion of Wolbachia in the CBB microbiota from 0.49% to 0.04%. This reduction affected CBB reproduction: females fed with tetracycline had significantly fewer progeny, lower fecundity, and fewer eggs per female. Tetracycline also reduced the population growth rate (λ), net reproductive rate (R0), and mean generation time (T) in CBB; the reduction in population growth was mostly due to variation in fertility, according to life time response experiments (LTREs) analysis. Our results suggest that Wolbachia contribute to the reproductive success of the CBB and their manipulation represents a possible approach to CBB biocontrol mediated by microbiome management.}, } @article {pmid28082006, year = {2017}, author = {Ilinsky, Y and Kosterin, OE}, title = {Molecular diversity of Wolbachia in Lepidoptera: Prevalent allelic content and high recombination of MLST genes.}, journal = {Molecular phylogenetics and evolution}, volume = {109}, number = {}, pages = {164-179}, doi = {10.1016/j.ympev.2016.12.034}, pmid = {28082006}, issn = {1095-9513}, mesh = {*Alleles ; Animals ; Butterflies/microbiology ; *Genes, Bacterial ; Genetic Loci ; *Genetic Variation ; Geography ; Lepidoptera/*microbiology ; *Multilocus Sequence Typing ; Nucleotides/genetics ; Phylogeny ; Recombination, Genetic/genetics ; Species Specificity ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia are common endosymbiotic bacteria of Arthropoda and Nematoda that are ordinarily transmitted vertically in host lineages through the egg cytoplasm. Despite the great interest in the Wolbachia symbiont, many issues of its biology remain unclear, including its evolutionary history, routes of transfer among species, and the molecular mechanisms underlying the symbiont's effect on its host. In this report, we present data relating to Wolbachia infection in 120 species of 13 Lepidoptera families, mostly butterflies, from West Siberian localities based on Multilocus sequence typing (MLST) and the wsp locus and perform a comprehensive survey of the distribution of Wolbachia and its genetic diversity in Lepidoptera worldwide. We observed a high infection incidence in the studied region; this finding is probably also true for other temperate latitude regions because many studied species have broad Palearctic and even Holarctic distribution. Although 40 new MLST alleles and 31 new STs were described, there was no noticeable difference in the MLST allele content in butterflies and probably also in moths worldwide. A genetic analysis of Wolbachia strains revealed the MLST allele core in lepidopteran hosts worldwide, viz. the ST-41 allele content. The key finding of our study was the detection of rampant recombination among MLST haplotypes. High rates of homologous recombination between Wolbachia strains indicate a substantial contribution of genetic exchanges to the generation of new STs. This finding should be considered when discussing issues related to the reconstruction of Wolbachia evolution, divergence time, and the routes of Wolbachia transmission across arthropod hosts.}, } @article {pmid28074180, year = {2016}, author = {Nakao, R and Abe, T and Funayama, S and Sugimoto, C}, title = {Horizontally Transferred Genetic Elements in the Tsetse Fly Genome: An Alignment-Free Clustering Approach Using Batch Learning Self-Organising Map (BLSOM).}, journal = {BioMed research international}, volume = {2016}, number = {}, pages = {3164624}, pmid = {28074180}, issn = {2314-6141}, mesh = {Animals ; Bacillus/genetics ; Chromosome Mapping/methods ; *Evolution, Molecular ; *Gene Transfer, Horizontal ; *Genetic Loci ; *Genome ; Tsetse Flies/*genetics/microbiology ; Wolbachia/genetics ; }, abstract = {Tsetse flies (Glossina spp.) are the primary vectors of trypanosomes, which can cause human and animal African trypanosomiasis in Sub-Saharan African countries. The objective of this study was to explore the genome of Glossina morsitans morsitans for evidence of horizontal gene transfer (HGT) from microorganisms. We employed an alignment-free clustering method, that is, batch learning self-organising map (BLSOM), in which sequence fragments are clustered based on the similarity of oligonucleotide frequencies independently of sequence homology. After an initial scan of HGT events using BLSOM, we identified 3.8% of the tsetse fly genome as HGT candidates. The predicted donors of these HGT candidates included known symbionts, such as Wolbachia, as well as bacteria that have not previously been associated with the tsetse fly. We detected HGT candidates from diverse bacteria such as Bacillus and Flavobacteria, suggesting a past association between these taxa. Functional annotation revealed that the HGT candidates encoded loci in various functional pathways, such as metabolic and antibiotic biosynthesis pathways. These findings provide a basis for understanding the coevolutionary history of the tsetse fly and its microbes and establish the effectiveness of BLSOM for the detection of HGT events.}, } @article {pmid28057823, year = {2017}, author = {McLaughlin, RN and Malik, HS}, title = {Genetic conflicts: the usual suspects and beyond.}, journal = {The Journal of experimental biology}, volume = {220}, number = {Pt 1}, pages = {6-17}, pmid = {28057823}, issn = {1477-9145}, support = {/HHMI/Howard Hughes Medical Institute/United States ; R01 GM074108/GM/NIGMS NIH HHS/United States ; K99 GM112941/GM/NIGMS NIH HHS/United States ; P50 GM107632/GM/NIGMS NIH HHS/United States ; R00 GM112941/GM/NIGMS NIH HHS/United States ; }, mesh = {Adaptation, Physiological ; Animals ; Biological Evolution ; DNA Transposable Elements ; *Evolution, Molecular ; Gene-Environment Interaction ; *Genome ; Host-Pathogen Interactions ; Humans ; Meiosis ; Models, Genetic ; Selection, Genetic ; }, abstract = {Selfishness is pervasive and manifests at all scales of biology, from societies, to individuals, to genetic elements within a genome. The relentless struggle to seek evolutionary advantages drives perpetual cycles of adaptation and counter-adaptation, commonly referred to as Red Queen interactions. In this review, we explore insights gleaned from molecular and genetic studies of such genetic conflicts, both extrinsic (between genomes) and intrinsic (within genomes or cells). We argue that many different characteristics of selfish genetic elements can be distilled into two types of advantages: an over-replication advantage (e.g. mobile genetic elements in genomes) and a transmission distortion advantage (e.g. meiotic drivers in populations). These two general categories may help classify disparate types of selfish genetic elements.}, } @article {pmid28056065, year = {2017}, author = {Ross, PA and Wiwatanaratanabutr, I and Axford, JK and White, VL and Endersby-Harshman, NM and Hoffmann, AA}, title = {Wolbachia Infections in Aedes aegypti Differ Markedly in Their Response to Cyclical Heat Stress.}, journal = {PLoS pathogens}, volume = {13}, number = {1}, pages = {e1006006}, pmid = {28056065}, issn = {1553-7374}, mesh = {Aedes/*microbiology ; Animals ; Environment ; Hot Temperature ; Insect Vectors/microbiology ; Pest Control, Biological/*methods ; Wolbachia/*growth & development ; }, abstract = {Aedes aegypti mosquitoes infected with Wolbachia bacteria are currently being released for arbovirus suppression around the world. Their potential to invade populations and persist will depend on interactions with environmental conditions, particularly as larvae are often exposed to fluctuating and extreme temperatures in the field. We reared Ae. aegypti larvae infected with different types of Wolbachia (wMel, wAlbB and wMelPop-CLA) under diurnal cyclical temperatures. Rearing wMel and wMelPop-CLA-infected larvae at 26-37°C reduced the expression of cytoplasmic incompatibility, a reproductive manipulation induced by Wolbachia. We also observed a sharp reduction in the density of Wolbachia in adults. Furthermore, the wMel and wMelPop-CLA infections were not transmitted to the next generation when mosquitoes were exposed to 26-37°C across all life stages. In contrast, the wAlbB infection was maintained at a high density, exhibited complete cytoplasmic incompatibility, and was transmitted from mother to offspring with a high fidelity under this temperature cycle. These findings have implications for the potential success of Wolbachia interventions across different environments and highlight the importance of temperature control in rearing.}, } @article {pmid28056021, year = {2017}, author = {Klarmann-Schulz, U and Specht, S and Debrah, AY and Batsa, L and Ayisi-Boateng, NK and Osei-Mensah, J and Mubarik, Y and Konadu, P and Ricchiuto, A and Fimmers, R and Arriens, S and Dubben, B and Ford, L and Taylor, M and Hoerauf, A}, title = {Comparison of Doxycycline, Minocycline, Doxycycline plus Albendazole and Albendazole Alone in Their Efficacy against Onchocerciasis in a Randomized, Open-Label, Pilot Trial.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {1}, pages = {e0005156}, pmid = {28056021}, issn = {1935-2735}, mesh = {Adolescent ; Adult ; Albendazole/*administration & dosage ; Animals ; Anthelmintics/*administration & dosage ; Doxycycline/*administration & dosage ; Drug Therapy, Combination ; Female ; Ghana ; Humans ; Male ; Middle Aged ; Minocycline/*administration & dosage ; Onchocerca volvulus/drug effects/genetics/isolation & purification/physiology ; Onchocerciasis/*drug therapy/parasitology ; Pilot Projects ; Treatment Outcome ; Young Adult ; }, abstract = {UNLABELLED: The search for new macrofilaricidal drugs against onchocerciasis that can be administered in shorter regimens than required for doxycycline (DOX, 200mg/d given for 4-6 weeks), identified minocycline (MIN) with superior efficacy to DOX. Further reduction in the treatment regimen may be achieved with co-administration with standard anti-filarial drugs. Therefore a randomized, open-label, pilot trial was carried out in an area in Ghana endemic for onchocerciasis, comprising 5 different regimens: the standard regimen DOX 200mg/d for 4 weeks (DOX 4w, N = 33), the experimental regimens MIN 200mg/d for 3 weeks (MIN 3w; N = 30), DOX 200mg/d for 3 weeks plus albendazole (ALB) 800mg/d for 3 days (DOX 3w + ALB 3d, N = 32), DOX 200mg/d for 3 weeks (DOX 3w, N = 31) and ALB 800mg for 3 days (ALB 3d, N = 30). Out of 158 randomized participants, 116 (74.4%) were present for the follow-up at 6 months of whom 99 participants (63.5%) followed the treatment per protocol and underwent surgery. Histological analysis of the adult worms in the extirpated nodules revealed absence of Wolbachia in 98.8% (DOX 4w), 81.4% (DOX 3w + ALB 3d), 72.7% (MIN 3w), 64.1% (DOX 3w) and 35.2% (ALB 3d) of the female worms. All 4 treatment regimens showed superiority to ALB 3d (p < 0.001, p < 0.001, p = 0.002, p = 0.008, respectively), which was confirmed by real-time PCR. Additionally, DOX 4w showed superiority to all other treatment arms. Furthermore DOX 4w and DOX 3w + ALB 3d showed a higher amount of female worms with degenerated embryogenesis compared to ALB 3d (p = 0.028, p = 0.042, respectively). These results confirm earlier studies that DOX 4w is sufficient for Wolbachia depletion and the desired parasitological effects. The data further suggest that there is an additive effect of ALB (3 days) on top of that of DOX alone, and that MIN shows a trend for stronger potency than DOX. These latter two results are preliminary and need confirmation in a fully randomized controlled phase 2 trial.

TRIAL REGISTRATION: ClinicalTrials.gov #06010453.}, } @article {pmid28052065, year = {2017}, author = {Joubert, DA and O'Neill, SL}, title = {Comparison of Stable and Transient Wolbachia Infection Models in Aedes aegypti to Block Dengue and West Nile Viruses.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {1}, pages = {e0005275}, pmid = {28052065}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Dengue/microbiology/transmission ; Dengue Virus/*physiology ; Female ; Insect Vectors ; Male ; Models, Theoretical ; Virus Replication/*physiology ; West Nile Fever/microbiology/transmission ; West Nile virus/*physiology ; Wolbachia/*physiology ; }, abstract = {Pathogen replication and transmission in Wolbachia infected insects are currently studied using three Wolbachia infection systems: naturally infected Wolbachia hosts, hosts transinfected with Wolbachia (stably maintained and inherited infections) and hosts transiently infected with Wolbachia. All three systems have been used to test the effect of Wolbachia on mosquito transmitted pathogens such as dengue virus (DENV), West Nile virus (WNV) and Plasmodium. From these studies it is becoming increasingly clear that the interaction between a particular pathogen and Wolbachia is heavily influenced by the host-Wolbachia interaction and the model of infection. In particular, there is some evidence that under very specific conditions, Wolbachia can enhance pathogen infection in some hosts. In this study, we compared the effect of Wolbachia in two infection models (stable transinfected and transiently infected) on the replication, infection- and transmission rates of two flaviviruses, DENV and WNV (Kunjin strain). Our results indicate that Wolbachia had similar blocking effects in both stable and transient models of infection, however, the magnitude of the blocking effect was significantly lower in mosquitoes transiently infected with Wolbachia. More importantly, no evidence was found for any enhancement of either DENV or WNV (Kunjin strain) infection in Ae. aegypti infected with Wolbachia, supporting a role for Wolbachia as an effective and safe means for restricting transmission of these viruses.}, } @article {pmid28049921, year = {2017}, author = {Takashima, Y and Onoda, I and Chiou, SP and Kitoh, K}, title = {In vitro canine platelet aggregation caused by Dirofilaria immitis extract.}, journal = {The Journal of veterinary medical science}, volume = {79}, number = {2}, pages = {387-392}, pmid = {28049921}, issn = {1347-7439}, mesh = {Animals ; Complex Mixtures/pharmacology ; Dirofilaria immitis/*pathogenicity ; *Dogs ; Female ; Hot Temperature ; In Vitro Techniques ; Male ; *Platelet Aggregation/drug effects ; Prostaglandin-Endoperoxide Synthases/metabolism ; Receptors, Purinergic P2/metabolism ; }, abstract = {Platelet function hyper-activity has been reported in Dirofilaria immitis (heartworm, HW)-infected dogs. Although the mechanism of increased platelet hyper-activity has not yet been elucidated, it is suggested to be mediated by unknown factors, which may be related to adult HW components. This study aims to determine whether adult male HW whole body extract induces canine platelet aggregation in vitro. The results indicate that HW extract caused an aggregation of canine platelets in a concentration-dependent manner. This aggregation ability of the HW extract was not mediated by the adenosine diphosphate receptor. In addition, the mechanisms of aggregation did not require cyclooxygenase-dependent pathways, and the aggregating activity of substances contained in the HW extract was heat stable; therefore, the active substances may be different from collagen. Furthermore, the platelet aggregating activity remained within the molecular weight (MW)≥100,000 fraction obtained by ultrafiltrating the HW extract. In contrast, the MW <100,000 fraction also had a platelet aggregation ability, but the aggregation pattern was reversible and the maximum extent decreased, compared with the MW≥100,000 fraction response. Our experiments have been conducted using a whole body extract from adult HWs to determine with certainty the aggregating activity of HW elements on canine platelets. More studies are necessary to evaluate the effects of the metabolic products released from live adult worms in pulmonary arteries and the symbiont bacterium Wolbachia-derived antigens on canine platelet aggregation.}, } @article {pmid28049478, year = {2017}, author = {Caragata, EP and Pais, FS and Baton, LA and Silva, JB and Sorgine, MH and Moreira, LA}, title = {The transcriptome of the mosquito Aedes fluviatilis (Diptera: Culicidae), and transcriptional changes associated with its native Wolbachia infection.}, journal = {BMC genomics}, volume = {18}, number = {1}, pages = {6}, pmid = {28049478}, issn = {1471-2164}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Computational Biology/methods ; Contig Mapping ; *Gene Expression Profiling ; Gene Expression Regulation ; Gene Ontology ; High-Throughput Nucleotide Sequencing ; Host-Pathogen Interactions/*genetics ; Molecular Sequence Annotation ; *Transcriptome ; *Wolbachia ; }, abstract = {BACKGROUND: Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquito Aedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection.

RESULTS: In total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes.

CONCLUSIONS: While the scale of transcriptional changes associated with wFlu infection might be small, the scope is rather large, which confirms that native Wolbachia infections maintain intricate molecular relationships with their mosquito hosts even after lengthy periods of co-evolution. We have also identified several potential means through which wFlu infection might influence Plasmodium infection in Ae. fluviatilis, and these genes should form the basis of future investigation into the enhancement of Plasmodium by Wolbachia.}, } @article {pmid30721615, year = {2017}, author = {Bogacheva, AS and Shaikevich, EV and Rakova, VM and Ganushkina, LA}, title = {[(THE FAUNA OF BLOODSUCKING MOSQUITOES IN THE NIZHNY NOVGOROD REGION, THEIR INFECTION WITH DIROFILARIA AND ENDOSYMBIOTIC BACTERIA)].}, journal = {Meditsinskaia parazitologiia i parazitarnye bolezni}, volume = {1}, number = {1}, pages = {43-47}, pmid = {30721615}, issn = {0025-8326}, mesh = {Aedes/microbiology/parasitology ; Animals ; DNA, Helminth/genetics ; Dirofilaria immitis/microbiology/pathogenicity ; Dirofilaria repens/microbiology/pathogenicity ; Dirofilariasis/*microbiology/*parasitology ; Dogs ; Humans ; Mosquito Vectors/genetics/*microbiology/*parasitology ; *Symbiosis ; Wolbachia/isolation & purification/pathogenicity ; }, abstract = {The fauna of bloodsucking mosquitoes in the Nizhny Novgorod Region is represented by 11 species from 5 genera of the family Culicidae. During 2014-2015, the predominant species were Ochlerotatus cantans and Aedes cinereus mosqui- toes in both a population aggregate and woodland. The infected mosquitoes accounted for 1.3% of their total number and were registered only in the village of Fokino. The investigators identified two human pathogenic nematode species: Diro- filaria immits and Dirofilaria repens (0.9% and 0.4% respectively). The effective carriers of Dirofilaria in the examined area can be Ae.cinereus and Och.cantans as only these species were found to have an invasive stage of the parasite. The symbiotic bacterium Wolbachia was detected in the mosquitoes that were not infected with dirofilariasis. This is the first study in Russia to investigate the effects of Wolbachia on the susceptibility of dirofilariasis vectors to infection.}, } @article {pmid28032096, year = {2016}, author = {Simsek, S and Ciftci, AT}, title = {Serological and Molecular Detection of Dirofilaria Species in Stray Dogs and Investigation of Wolbachia DNA by PCR in Turkey.}, journal = {Journal of arthropod-borne diseases}, volume = {10}, number = {4}, pages = {445-453}, pmid = {28032096}, issn = {2322-1984}, abstract = {BACKGROUND: Dirofilaria immitis and Dirofilaria repens are the most common species of filarial nematodes described in the dogs. A single-step multiplex PCR was applied to detect and differentiate simultaneously and unequivocally D. immitis and D. repens on DNA extracted from canine peripheral blood and besides to detect the seroprevalance of D. immitis by ELISA in Elazig Province, Turkey. A PCR detection of the Wolbachia, which plays an important role in D. immitis biology and contributes to the inflammatory pathology of the heartworm, was also applied for the first time in Turkey.

METHODS: A total of 161 whole blood and sera samples were collected from stray dogs and stored at -20 °C until used. After DNA extraction, all samples were processed with Dirofilaria primers by multiplex-PCR and Wolbachia primers by conventional PCR besides ELISA for serology. The amplification was performed using a set of primers designed on a portion of the small subunit ribosomal RNA gene of the mitochondrion (12S rDNA).

RESULTS: Three of the examined dogs (1.8%) were found to be infected with only D. immitis, one (0.6%) with D. repens and three (1.8%) with both parasites. Besides, 10 out of 161 dogs (6.2%) were found infected with Wolbachia sp. Finaly, the seroprevalence of dirofilariosis in the examined dogs was found to be 3.7% (6/161).

CONCLUSION: Although dirofilariosis is not a serious problem in the region, the stray dogs still continue to be a source of infection.}, } @article {pmid28028309, year = {2017}, author = {Hofer, U}, title = {Symbiosis: New horizons for Wolbachia.}, journal = {Nature reviews. Microbiology}, volume = {15}, number = {2}, pages = {66-67}, pmid = {28028309}, issn = {1740-1534}, mesh = {Genome, Bacterial ; Sex Chromosomes ; Symbiosis ; *Wolbachia ; }, } @article {pmid28028089, year = {2016}, author = {Schuler, H and Kern, P and Arthofer, W and Vogt, H and Fischer, M and Stauffer, C and Riegler, M}, title = {Wolbachia in Parasitoids Attacking Native European and Introduced Eastern Cherry Fruit Flies in Europe.}, journal = {Environmental entomology}, volume = {45}, number = {6}, pages = {1424-1431}, doi = {10.1093/ee/nvw137}, pmid = {28028089}, issn = {1938-2936}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Austria ; DNA, Bacterial/genetics ; Electron Transport Complex IV/genetics ; Germany ; Insect Proteins/genetics ; Introduced Species ; Multilocus Sequence Typing ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Species Specificity ; Tephritidae/microbiology/*parasitology ; Wasps/*microbiology/*physiology ; Wolbachia/genetics/*physiology ; }, abstract = {The eastern cherry fruit fly, Rhagoletis cingulata Loew (Diptera: Tephritidae), is an economically important pest of cherries in North America. In 1983 it was first reported in Europe where it shares its ecological niche with the native European cherry fruit fly, Rhagoletis cerasi L. (Diptera: Tephritidae). Their coexistence in Europe led to the recent horizontal transmission of the Wolbachia strain wCer1 from R. cerasi to R. cingulata Horizontal Wolbachia transmission is mediated by either sharing of ecological niches or by interacting species such as parasitoids. Here we describe for the first time that two braconid wasps, Psyttalia rhagoleticola Sachtleben (Hymenoptera: Braconidae) and Utetes magnus Fischer (Hymenoptera: Braconidae), naturally parasitizing R. cerasi, use the invasive R. cingulata in Europe as a new host. In contrast, no parasitoids that parasitize R. cingulata in its native American range were detected in the introduced European range. Diagnostic Wolbachia PCR screening and sequence analyses demonstrated that all P. rhagoleticola individuals were infected with the newly described Wolbachia strain wRha while all U. magnus individuals were uninfected. wRha is different from wCer1 but had an Wolbachia surface protein (wsp) gene sequence that was identical to wCer2 of R. cerasi and wCin2 of R. cingulata. However, multi locus sequence typing revealed differences in all loci between wRha and the tephritid's strains. The horizontal transmission of wCer1 between the two tephritid species did not result in fixed heritable infections in the parasitoids. However, the parasitoids may have acted as a transient wCer1 vector.}, } @article {pmid28026036, year = {2017}, author = {Dutra, HL and Caragata, EP and Moreira, LA}, title = {The re-emerging arboviral threat: Hidden enemies: The emergence of obscure arboviral diseases, and the potential use of Wolbachia in their control.}, journal = {BioEssays : news and reviews in molecular, cellular and developmental biology}, volume = {39}, number = {2}, pages = {}, doi = {10.1002/bies.201600175}, pmid = {28026036}, issn = {1521-1878}, mesh = {Animals ; Arbovirus Infections/*therapy ; Arthropod Vectors/*microbiology ; Biological Control Agents/*therapeutic use ; Humans ; *Wolbachia ; }, abstract = {Mayaro, Oropouche, and O'Nyong-Nyong share many traits with more prominent arboviruses, like dengue and yellow fever, chikungunya, and Zika. These include severe clinical symptoms, multiple animal hosts, and widespread vector species living in close proximity to human habitats, all of which constitute significant risk factors for more frequent outbreaks in the future, greatly increasing the potential of these hidden enemies to follow Zika and become the next wave of global arboviral threats. Critically, the current dearth of knowledge on these arboviruses might impede the success of future control efforts, including the potential application of Wolbachia pipientis. This bacterium inherently possesses broad anti-pathogen properties and a means of genetic drive that allows it to eliminate or replace target vector populations. We conclude that control of obscure arboviruses with Wolbachia might be possible, but successful implementation will be critically dependent on the ability to transinfect key vector species.}, } @article {pmid28009851, year = {2016}, author = {Gonzales, KK and Hansen, IA}, title = {Artificial Diets for Mosquitoes.}, journal = {International journal of environmental research and public health}, volume = {13}, number = {12}, pages = {}, pmid = {28009851}, issn = {1660-4601}, support = {R25 GM061222/GM/NIGMS NIH HHS/United States ; SC1 AI109055/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*growth & development ; Animals ; Diet/*methods ; Female ; Humans ; Insect Vectors/*growth & development ; *Research ; }, abstract = {Mosquito-borne diseases are responsible for more than a million human deaths every year. Modern mosquito control strategies such as sterile insect technique (SIT), release of insects carrying a dominant lethal (RIDL), population replacement strategies (PR), and Wolbachia-based strategies require the rearing of large numbers of mosquitoes in culture for continuous release over an extended period of time. Anautogenous mosquitoes require essential nutrients for egg production, which they obtain through the acquisition and digestion of a protein-rich blood meal. Therefore, mosquito mass production in laboratories and other facilities relies on vertebrate blood from live animal hosts. However, vertebrate blood is expensive to acquire and hard to store for longer times especially under field conditions. This review discusses older and recent studies that were aimed at the development of artificial diets for mosquitoes in order to replace vertebrate blood.}, } @article {pmid28005061, year = {2016}, author = {Gerth, M and Bleidorn, C}, title = {Comparative genomics provides a timeframe for Wolbachia evolution and exposes a recent biotin synthesis operon transfer.}, journal = {Nature microbiology}, volume = {2}, number = {}, pages = {16241}, doi = {10.1038/nmicrobiol.2016.241}, pmid = {28005061}, issn = {2058-5276}, abstract = {The genus Wolbachia (Alphaproteobacteria) comprises the most abundant inherited intracellular bacteria[1]. Despite their relevance as manipulators of human pathogen transmission[2] and arthropod reproduction[3], many aspects of their evolutionary history are not well understood[4]. In arthropods, Wolbachia infections are typically transient on evolutionary timescales[5,6] and co-divergence between hosts and Wolbachia is supposedly rare. Consequently, much of our knowledge of Wolbachia genome evolution derives from very recently diverged strains, and a timescale for Wolbachia is lacking. Here, we investigated the genomes of four Wolbachia strains that have persisted within and co-diverged with their host lineage for ∼2 million years. Although the genomes showed very little evolutionary change on a nucleotide level, we found evidence for a recent lateral transfer of a complete biotin synthesis operon that has the potential to transform Wolbachia-host relationships[7]. Furthermore, this evolutionary snapshot enabled us to calibrate the divergence times of the supergroup A and B Wolbachia lineages using genome-wide data sets and relaxed molecular clock models. We estimated the origin of Wolbachia supergroups A and B to be ∼200 million years ago (Ma), which is considerably older than previously appreciated. This age coincides with the diversification of many insect lineages[8] that represent most of Wolbachia's host spectrum.}, } @article {pmid28004792, year = {2016}, author = {Ajendra, J and Specht, S and Ziewer, S and Schiefer, A and Pfarr, K and Parčina, M and Kufer, TA and Hoerauf, A and Hübner, MP}, title = {NOD2 dependent neutrophil recruitment is required for early protective immune responses against infectious Litomosoides sigmodontis L3 larvae.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {39648}, pmid = {28004792}, issn = {2045-2322}, mesh = {Acetylmuramyl-Alanyl-Isoglutamine/pharmacology ; Animals ; Cell Wall/immunology ; Cytokines/immunology ; Female ; Filariasis/*immunology ; Filarioidea ; Gene Expression Profiling ; HEK293 Cells ; Humans ; Immune System ; Larva ; Ligands ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred NOD ; Mice, Knockout ; Microbiota ; *Neutrophil Infiltration ; Neutrophils/immunology ; Nod2 Signaling Adaptor Protein/*immunology ; Signal Transduction ; Skin/microbiology/parasitology ; Thoracic Cavity/parasitology ; }, abstract = {Nucleotide-binding oligomerization domain-containing protein 2 (NOD2) recognizes muramyl dipeptide (MDP) of bacterial cell walls, triggering NFκB-induced pro-inflammation. As most human pathogenic filariae contain Wolbachia endobacteria that synthesize the MDP-containing cell wall precursor lipid II, NOD2's role during infection with the rodent filaria Litomosoides sigmodontis was investigated. In NFκB reporter-cells, worm-extract containing Wolbachia induced NOD2 and NOD1. NOD2-deficient mice infected with L. sigmodontis had significantly more worms than wildtype controls early in infection. Increased worm burden was not observed after subcutaneous infection, suggesting that protective NOD2-dependent immune responses occur within the skin. Flow cytometry demonstrated that neutrophil recruitment to the skin was impaired in NOD2[-/-] mice after intradermal injection of third stage larvae (L3), and blood neutrophil numbers were reduced after L. sigmodontis infection. PCR array supported the requirement of NOD2 for recruitment of neutrophils to the skin, as genes associated with neutrophil recruitment and activation were downregulated in NOD2[-/-] mice after intradermal L3 injection. Neutrophil depletion before L. sigmodontis infection increased worm recovery in wildtype mice, confirming that neutrophils are essential against invading L3 larvae. This study indicates that NOD-like receptors are implemented in first-line protective immune responses against filarial nematodes.}, } @article {pmid27990558, year = {2017}, author = {Gill, TA and Chu, C and Pelz-Stelinski, KS}, title = {Comparative proteomic analysis of hemolymph from uninfected and Candidatus Liberibacter asiaticus-infected Diaphorina citri.}, journal = {Amino acids}, volume = {49}, number = {2}, pages = {389-406}, doi = {10.1007/s00726-016-2373-2}, pmid = {27990558}, issn = {1438-2199}, mesh = {Animals ; Hemiptera/*microbiology ; Hemolymph/*metabolism/microbiology ; Host-Pathogen Interactions ; Insect Proteins/*analysis/metabolism ; Proteomics/*methods ; Rhizobiaceae/*pathogenicity ; Symbiosis ; Wolbachia/metabolism ; }, abstract = {Hemolymph was characterized from Diaphorina citri adults infected with the phytopathogen, Candidatus Liberibacter asiaticus (CLas), and compared with that from uninfected psyllids. This study identified 5531 and 3220 peptides within infected and uninfected hemolymph using nano-LC-MS/MS. A reduced number of proteins were detected for D. citri and all known endosymbionts within infected hemolymph as compared to uninfected hemolymph. A large number of immune defense proteins were absent from D. citri hemolymph; however, a single recognition protein (PGRP), two serine protease inhibitors, three prophenoloxidase (proPO) enzymes, and a single serine protease in an uninfected D. citri were detected. The hemolymph is nearly devoid of nutrient storage proteins. This is the first proteomic analysis of D. citri hemolymph that also analyses the components contributed by all the endosymbionts. By comparing the contribution of each endosymbiont (CCR, CPA, and WB) in the presence and absence of CLas infection, this study offers initial insights regarding the hemolymph response to microbial community shifts associated with D. citri infection status. Our data also present potential protein targets for analysis and disruption of CLas transmission that may facilitate management of huanglongbing (HLB) caused by CLas in citrus.}, } @article {pmid27988830, year = {2017}, author = {Espinosa, MS and Virla, EG and Cuozzo, S}, title = {Wolbachia Infections Responsible for Thelytoky in Dryinid Wasps. The Case of Gonatopus bonaerensis Virla (Hymenoptera: Dryinidae).}, journal = {Neotropical entomology}, volume = {46}, number = {4}, pages = {409-413}, pmid = {27988830}, issn = {1678-8052}, mesh = {Animals ; Female ; Male ; *Parthenogenesis ; Reproduction ; Wasps/*microbiology ; *Wolbachia/genetics/pathogenicity ; }, abstract = {We studied the occurrence of Wolbachia in the parasitoid Gonatopus bonaerensis Virla (Hymenoptera: Dryinidae). In order to verify the existence of natural infections in the parasitoid, a field survey was conducted. Identification of Wolbachia was performed on the basis of 16S rDNA, wsp_F1, and wsp_R1-sequences. After the detection of the bacteria, infected specimens of G. bonaerensis were treated with a solution of tetracycline. In Tucumán, parasitoids hold Wolbachia endosymbiont, which seems to control the wasp's reproduction in the nature turning it into thelytokous. The symbiont was identified as the Wolbachia sp. wRi strain. The cure of infected unfertilized females determined the normal arrhenotokous parthenogenesis and the production of male offspring. As a consequence of this procedure, the male of G. bonaerensis is described for the first time.}, } @article {pmid27982076, year = {2016}, author = {Zhou, XF and Li, ZX}, title = {Establishment of the cytoplasmic incompatibility-inducing Wolbachia strain wMel in an important agricultural pest insect.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {39200}, pmid = {27982076}, issn = {2045-2322}, mesh = {Animals ; DNA, Bacterial/chemistry/isolation & purification/metabolism ; Drosophila melanogaster/*microbiology ; Female ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Multilocus Sequence Typing ; Ovum/microbiology ; *Pest Control, Biological ; Sequence Analysis, DNA ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {The wMel Wolbachia strain was known for cytoplasmic incompatibility (CI)-induction and blocking the transmission of dengue. However, it is unknown whether it can establish and induce CI in a non-dipteran host insect. Here we artificially transferred wMel from Drosophila melanogaster into the whitefly Bemisia tabaci. Fluorescence in situ hybridisation demonstrated that wMel had successfully transfected the new host. Reciprocal crossing was conducted with wMel-transfected and wild-type isofemale lines, indicating that wMel could induce a strong CI without imposing significant cost on host fecundity. We then determined the maternal transmission efficiency of wMel in the offspring generations, showing a fluctuating trend over a period of 12 generations. We thus detected the titre of wMel during different developmental stages and in different generations by using real-time quantitative PCR, revealing a similar fluctuating mode, but it was not significantly correlated with the dynamics of transmission efficiency. These results suggest that wMel can be established in B.tabaci, a distantly related pest insect of agricultural importance; moreover, it can induce a strong CI phenotype in the recipient host insect, suggesting a potential for its use in biological control of B. tabaci.}, } @article {pmid27973604, year = {2016}, author = {Chaplinska, M and Gerritsma, S and Dini-Andreote, F and Falcao Salles, J and Wertheim, B}, title = {Bacterial Communities Differ among Drosophila melanogaster Populations and Affect Host Resistance against Parasitoids.}, journal = {PloS one}, volume = {11}, number = {12}, pages = {e0167726}, pmid = {27973604}, issn = {1932-6203}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Bacteria/*classification ; Disease Resistance/*genetics ; Drosophila melanogaster/*microbiology/*parasitology ; Feeding Behavior ; Female ; Founder Effect ; Genetics, Population ; Genotype ; Geography ; Host-Parasite Interactions ; Larva/parasitology ; Microbial Consortia ; *Microbiota ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Species Specificity ; Wasps/physiology ; Wolbachia ; }, abstract = {In Drosophila, diet is considered a prominent factor shaping the associated bacterial community. However, the host population background (e.g. genotype, geographical origin and founder effects) is a factor that may also exert a significant influence and is often overlooked. To test for population background effects, we characterized the bacterial communities in larvae of six genetically differentiated and geographically distant D. melanogaster lines collected from natural populations across Europe. The diet for these six lines had been identical for ca. 50 generations, thus any differences in the composition of the microbiome originates from the host populations. We also investigated whether induced shifts in the microbiome-in this case by controlled antibiotic administration-alters the hosts' resistance to parasitism. Our data revealed a clear signature of population background on the diversity and composition of D. melanogaster microbiome that differed across lines, even after hosts had been maintained at the same diet and laboratory conditions for over 4 years. In particular, the number of bacterial OTUs per line ranged from 8 to 39 OTUs. Each line harboured 2 to 28 unique OTUs, and OTUs that were highly abundant in some lines were entirely missing in others. Moreover, we found that the response to antibiotic treatment differed among the lines and significantly altered the host resistance to the parasitoid Asobara tabida in one of the six lines. Wolbachia, a widespread intracellular endosymbiont associated with parasitoid resistance, was lacking in this line, suggesting that other components of the Drosophila microbiome caused a change in host resistance. Collectively, our results revealed that lines that originate from different population backgrounds show significant differences in the established Drosophila microbiome, outpacing the long-term effect of diet. Perturbations on these naturally assembled microbiomes to some degree influenced the hosts' resistance against natural parasites.}, } @article {pmid27966221, year = {2017}, author = {Duan, D and Cheng, T}, title = {Determination of the microbial community features of Haemaphysalis flava in different developmental stages by high-throughput sequencing.}, journal = {Journal of basic microbiology}, volume = {57}, number = {4}, pages = {302-308}, doi = {10.1002/jobm.201600557}, pmid = {27966221}, issn = {1521-4028}, mesh = {Acinetobacter/genetics/isolation & purification ; Adult ; Animals ; Bacteria/*classification/*genetics/isolation & purification ; Coxiella/genetics/isolation & purification ; DNA, Bacterial/genetics ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Ixodidae/*growth & development/*microbiology ; Larva/microbiology ; Microbial Consortia/*genetics ; Nymph/microbiology ; Ovum/microbiology ; Phylogeny ; Pseudomonas/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; }, abstract = {In this study, we analyzed the in vivo microbial community structure and diversity differences of Haemaphysalis flava (H. flava) in four developmental stages (egg, larva, nymph, and adult) to determine which bacterial genera could be propagated through transovarial transmission and transmitted by transstadial transmission during different developmental stages of H. flava. Paired-end sequencing of the V3 region of the 16S ribosomal DNA (16S rDNA) of H. flava in four developmental stage samples was conducted using the Illumina MiSeq sequencing platform. The following operational taxonomic units (OTUs) of H. flava were obtained during the four developmental stages: 89 of egg, 111 of larva, 104 of nymph, and 106 of female adult tick. Sixty-four of these OTUs had high similarity in the four developmental stages of H. flava. Eight bacterial genera had the highest abundances in all developmental stages, namely, Rickettsia, Coxiella, Pseudomonas, Ehrlichia, Escherichia, Acinetobacter, Citrobacter, and Cupriavidus. The nymph had the highest abundance of Coxiella, and the female adult tick had the highest abundance of Rickettsia. Staphylococcus and Wolbachia were detected in all developmental stages except the egg.}, } @article {pmid27965627, year = {2016}, author = {Wang, GH and Sun, BF and Xiong, TL and Wang, YK and Murfin, KE and Xiao, JH and Huang, DW}, title = {Bacteriophage WO Can Mediate Horizontal Gene Transfer in Endosymbiotic Wolbachia Genomes.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1867}, pmid = {27965627}, issn = {1664-302X}, support = {T32 HL007974/HL/NHLBI NIH HHS/United States ; }, abstract = {Phage-mediated horizontal gene transfer (HGT) is common in free-living bacteria, and many transferred genes can play a significant role in their new bacterial hosts. However, there are few reports concerning phage-mediated HGT in endosymbionts (obligate intracellular bacteria within animal or plant hosts), such as Wolbachia. The Wolbachia-infecting temperate phage WO can actively shift among Wolbachia genomes and has the potential to mediate HGT between Wolbachia strains. In the present study, we extend previous findings by validating that the phage WO can mediate transfer of non-phage genes. To do so, we utilized bioinformatic, phylogenetic, and molecular analyses based on all sequenced Wolbachia and phage WO genomes. Our results show that the phage WO can mediate HGT between Wolbachia strains, regardless of whether the transferred genes originate from Wolbachia or other unrelated bacteria.}, } @article {pmid27935594, year = {2017}, author = {Li, SJ and Ahmed, MZ and Lv, N and Shi, PQ and Wang, XM and Huang, JL and Qiu, BL}, title = {Plantmediated horizontal transmission of Wolbachia between whiteflies.}, journal = {The ISME journal}, volume = {11}, number = {4}, pages = {1019-1028}, pmid = {27935594}, issn = {1751-7370}, mesh = {Animals ; Cucumis sativus/microbiology ; Hemiptera/*microbiology ; Host-Pathogen Interactions ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Vigna/microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Maternal transmission is the main transmission pathway of facultative bacterial endosymbionts, but phylogenetically distant insect hosts harbor closely related endosymbionts, suggesting that horizontal transmission occurs in nature. Here we report the first case of plant-mediated horizontal transmission of Wolbachia between infected and uninfected Bemisia tabaci AsiaII7 whiteflies. After infected whiteflies fed on cotton leaves, Wolbachia was visualized, both in the phloem vessels and in some novel 'reservoir' spherules along the phloem by fluorescence in situ hybridization using Wolbachia-specific 16S rRNA probes and transmission electron microscopy. Wolbachia persisted in the plant leaves for at least 50 days. When the Wolbachia-free whiteflies fed on the infected plant leaves, the majority of them became infected with the symbiont and vertically transmitted it to their progeny. Multilocus sequence typing and sequencing of the wsp (Wolbachia surface protein) gene confirmed that the sequence type of Wolbachia in the donor whiteflies, cotton phloem and the recipient whiteflies are all identical (sequence type 388). These results were replicated using cowpea and cucumber plants, suggesting that horizontal transmission is also possible through other plant species. Our findings may help explain why Wolbachia bacteria are so abundant in arthropods, and suggest that in some species, Wolbachia may be maintained in populations by horizontal transmission.}, } @article {pmid27934910, year = {2016}, author = {Guo, WP and Tian, JH and Lin, XD and Ni, XB and Chen, XP and Liao, Y and Yang, SY and Dumler, JS and Holmes, EC and Zhang, YZ}, title = {Extensive genetic diversity of Rickettsiales bacteria in multiple mosquito species.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {38770}, pmid = {27934910}, issn = {2045-2322}, mesh = {Animals ; Culicidae/*microbiology ; DNA, Bacterial/genetics ; *Genetic Variation ; Phylogeny ; Polymerase Chain Reaction ; Rickettsia/classification/*genetics ; Species Specificity ; }, abstract = {Rickettsiales are important zoonotic pathogens, causing severe disease in humans globally. Although mosquitoes are an important vector for diverse pathogens, with the exception of members of the genus Wolbachia little is known about their role in the transmission of Rickettsiales. Herein, Rickettsiales were identified by PCR in five species of mosquitoes (Anopheles sinensis, Armigeres subalbatus, Aedes albopictus, Culex quinquefasciatus and Cu. tritaeniorhynchus) collected from three Chinese provinces during 2014-2015. Subsequent phylogenetic analyses of the rrs, groEL and gltA genes revealed the presence of Anaplasma, Ehrlichia, Candidatus Neoehrlichia, and Rickettsia bacteria in mosquitoes, comprising nine documented and five tentative species bacteria, as well as three symbionts/endosybionts. In addition, bacteria were identified in mosquito eggs, larvae, and pupae sampled from aquatic environments. Hence, these data suggest that Rickettsiales circulate widely in mosquitoes in nature. Also of note was that Ehrlichia and Rickettsia bacteria were detected in each life stage of laboratory cultured mosquitoes, suggesting that Rickettsiales may be maintained in mosquitoes through both transstadial and transovarial transmission. In sum, these data indicate that mosquitoes may have played an important role in the transmission and evolution of Rickettsiales in nature.}, } @article {pmid27930295, year = {2016}, author = {Leclercq, S and Thézé, J and Chebbi, MA and Giraud, I and Moumen, B and Ernenwein, L and Grève, P and Gilbert, C and Cordaux, R}, title = {Birth of a W sex chromosome by horizontal transfer of Wolbachia bacterial symbiont genome.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {113}, number = {52}, pages = {15036-15041}, pmid = {27930295}, issn = {1091-6490}, mesh = {Animals ; Biological Evolution ; Crosses, Genetic ; Cytoplasm/metabolism ; Female ; *Gene Transfer, Horizontal ; *Genome, Bacterial ; Genotype ; Isopoda/microbiology ; Male ; Microscopy, Electron, Transmission ; Phylogeny ; *Sex Chromosomes ; Sex Determination Processes ; Sex Ratio ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {Sex determination is a fundamental developmental pathway governing male and female differentiation, with profound implications for morphology, reproductive strategies, and behavior. In animals, sex differences between males and females are generally determined by genetic factors carried by sex chromosomes. Sex chromosomes are remarkably variable in origin and can differ even between closely related species, indicating that transitions occur frequently and independently in different groups of organisms. The evolutionary causes underlying sex chromosome turnover are poorly understood, however. Here we provide evidence indicating that Wolbachia bacterial endosymbionts triggered the evolution of new sex chromosomes in the common pillbug Armadillidium vulgare We identified a 3-Mb insert of a feminizing Wolbachia genome that was recently transferred into the pillbug nuclear genome. The Wolbachia insert shows perfect linkage to the female sex, occurs in a male genetic background (i.e., lacking the ancestral W female sex chromosome), and is hemizygous. Our results support the conclusion that the Wolbachia insert is now acting as a female sex-determining region in pillbugs, and that the chromosome carrying the insert is a new W sex chromosome. Thus, bacteria-to-animal horizontal genome transfer represents a remarkable mechanism underpinning the birth of sex chromosomes. We conclude that sex ratio distorters, such as Wolbachia endosymbionts, can be powerful agents of evolutionary transitions in sex determination systems in animals.}, } @article {pmid27920393, year = {2017}, author = {Ahmad, NA and Vythilingam, I and Lim, YAL and Zabari, NZAM and Lee, HL}, title = {Detection of Wolbachia in Aedes albopictus and Their Effects on Chikungunya Virus.}, journal = {The American journal of tropical medicine and hygiene}, volume = {96}, number = {1}, pages = {148-156}, pmid = {27920393}, issn = {1476-1645}, mesh = {Aedes/*microbiology/*virology ; Animals ; Chikungunya virus/*isolation & purification ; Female ; Male ; Mosquito Control ; Phylogeny ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia-based vector control strategies have been proposed as a means to augment the currently existing measures for controlling dengue and chikungunya vectors. Prior to utilizing Wolbachia as a novel vector control strategy, it is crucial to understand the Wolbachia-mosquito interactions. In this study, field surveys were conducted to screen for the infection status of Wolbachia in field-collected Aedes albopictus The effects of Wolbachia in its native host toward the replication and dissemination of chikungunya virus (CHIKV) was also studied. The prevalence of Wolbachia-infected field-collected Ae. albopictus was estimated to be 98.6% (N = 142) for females and 95.1% (N = 102) for males in the population studied. The Ae. albopictus were naturally infected with both wAlbA and wAlbB strains. We also found that the native Wolbachia has no impact on CHIKV infection and minimal effect on CHIKV dissemination to secondary organs.}, } @article {pmid27914738, year = {2017}, author = {Prezotto, LF and Perondini, AL and Hernández-Ortiz, V and Marino, CL and Selivon, D}, title = {Wolbachia strains in cryptic species of the Anastrepha fraterculus complex (Diptera, Tephritidae) along the Neotropical Region.}, journal = {Systematic and applied microbiology}, volume = {40}, number = {1}, pages = {59-67}, doi = {10.1016/j.syapm.2016.11.002}, pmid = {27914738}, issn = {1618-0984}, mesh = {Americas ; Animals ; Cluster Analysis ; *Genetic Variation ; Genotype ; Molecular Typing ; Phylogeny ; Recombination, Genetic ; Tephritidae/*microbiology ; Tropical Climate ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Infection by Wolbachia was described previously in eleven species of Anastrepha fruit flies some of which are important pests of fruticulture. One such species is the nominal Anastrepha fraterculus, the South American fruit fly, which actually comprises a complex of cryptic species. The suggestions of using Wolbachia for the control of these pest species, make imperative a more precise characterization of the existing strains of the bacteria. In this study, population samples of the A. fraterculus complex from Brazil, Argentina, Peru, Ecuador, Colombia, Guatemala and Mexico were analyzed for Wolbachia infection. The bacteria were genotyped by the MLST and WSP Typing methodologies. All samples were infected with Wolbachia of supergroup "A". For each of the five MLST genes, unique as well as already known alleles were detected. Nineteen sequence types for the concatenated sequences of the five MLST genes, and twenty wsp alleles were found in the samples. Host-specific haplotypes, shared strains among distinct hosts, and more than one strain of Wolbachia were found in some population samples. Recombination among the MLST genes and intragenic recombination between wsp haplotypes was rare. Phylogenetic analysis showed a great similarity among the Wolbachia strains in the A. fraterculus complex. However, some strains of Wolbachia are found throughout the Neotropical Region and there are specific strains in determined geographical areas.}, } @article {pmid27905665, year = {2017}, author = {Valette, V and Durand, S and Bech, N and Grandjean, F and Beltran-Bech, S}, title = {Multiple paternity in a wild population of Armadillidium vulgare: influence of infection with Wolbachia?.}, journal = {Journal of evolutionary biology}, volume = {30}, number = {2}, pages = {235-243}, doi = {10.1111/jeb.13009}, pmid = {27905665}, issn = {1420-9101}, mesh = {Animals ; Female ; Isopoda/*parasitology ; Male ; Microsatellite Repeats ; *Paternity ; *Sex Ratio ; Wolbachia/*pathogenicity ; }, abstract = {Female multiple mating has been extensively studied to understand how nonobvious benefits, generally thought to be of genetic nature, could overcome heavy costs such as an increased risk of infection during mating. However, the impact of infection itself on multiple mating has rarely been addressed. The interaction between the bacterium Wolbachia and its terrestrial crustacean host, Armadillidium vulgare, is a relevant model to investigate this question. In this association, Wolbachia is able to turn genetic males into functional females (i.e. feminization), thereby distorting the sex ratio and decreasing the number of available males at the population scale. Moreover, in A. vulgare, females have been shown to mate multiply under laboratory conditions and males prefer uninfected females over infected ones. Additionally, different Wolbachia strains are known to infect A. vulgare and these strains differ in their transmission rate and virulence. All these elements suggest a potential impact of different Wolbachia strains on multiple mating. To investigate this assumption, we collected gravid females in a wild A. vulgare population harbouring both uninfected females and females infected with one of two different Wolbachia strains (wVulM and wVulC) and performed paternity analyses on the obtained broods using microsatellite markers. We demonstrate that (i) multiple paternity is common in this wild population of A. vulgare, with a mean number of fathers of 4.48 ± 1.24 per brood and (ii) females infected with wVulC produced broods with a lower multiple paternity level compared with females infected with wVulM and uninfected ones. This work improves our knowledge of the impact of infections on reproductive strategies.}, } @article {pmid27902358, year = {2016}, author = {Zhang, G and Etebari, K and Asgari, S}, title = {Wolbachia suppresses cell fusing agent virus in mosquito cells.}, journal = {The Journal of general virology}, volume = {97}, number = {12}, pages = {3427-3432}, doi = {10.1099/jgv.0.000653}, pmid = {27902358}, issn = {1465-2099}, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Cell Fusion ; Cell Line ; Flavivirus/*physiology ; *Virus Replication ; Wolbachia/*physiology ; }, abstract = {The genus Flavivirus contains a large number of positive-sense ssRNA viruses. While some are transmitted by mosquitoes or other arthropods and are pathogenic to humans and animals (e.g. dengue and Zika viruses), some are insect-specific and do not replicate in vertebrate cells. These are known as insect-specific flaviviruses (ISFs). Cell fusing agent virus (CFAV) was the first described ISF, which was detected in an Aedes aegypti cell line, Aag2. Here, we investigated the effect of Wolbachia, a widespread endosymbiont of many insect species, that is known to block replication of several pathogenic flaviviruses, on CFAV. Our results demonstrated that, in mosquito cells, Wolbachia vastly suppresses replication of CFAV, with significantly less CFAV viral interfering small RNAs produced in the cells. However, removal of Wolbachia with tetracycline led to increased CFAV replication. These results suggest that Wolbachia is also able to suppress an ISF.}, } @article {pmid27902186, year = {2017}, author = {Cicala, F and Moore, JD and Cáceres-Martínez, J and Del Río-Portilla, MA and Hernández-Rodríguez, M and Vásquez-Yeomans, R and Rocha-Olivares, A}, title = {Multigenetic characterization of 'Candidatus Xenohaliotis californiensis'.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {67}, number = {1}, pages = {42-49}, doi = {10.1099/ijsem.0.001563}, pmid = {27902186}, issn = {1466-5034}, mesh = {Anaplasmataceae/*classification/genetics ; Animals ; Bacterial Typing Techniques ; Bayes Theorem ; DNA, Bacterial/genetics ; Gastropoda/*microbiology ; Genes, Bacterial ; Likelihood Functions ; Models, Genetic ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {'Candidatus Xenohaliotis californiensis' (or Ca.Xc) is the aetiological agent of withering syndrome, a chronic wasting disease affecting most if not all North American species of abalone, and has been described as a Rickettsiales-like prokaryote. Genetic data regarding this species are limited to the 16S rRNA gene. The inability to grow it axenically has hindered its genetic and genomic characterization and, in consequence, a thorough analysis of its systematics. Here, we amplified and sequenced five genes (16S rRNA, 23S rRNA, ftsZ, virD4 and virB11) of Ca.Xc from infected abalone to analyse its phylogenetic position. Phylogenies from concatenated DNA and amino acid sequences with representative genera of most Rickettsiales unequivocally place Ca.Xc in the family Anaplasmataceae. Furthermore, the family has two reciprocally monophyletic lineages: one leading to (Neorickettsia, Ca.Xc) and the other to ((Ehrlichia, Anaplasma), Wolbachia)). A molecular-clock Bayesian reconstruction places Ca.Xc as the most basal lineage in Anaplasmataceae. These phylogenetic hypotheses shed light on patterns of host evolution and of ecological transitions. Specifically, Neorickettsia and Ca.Xc inhabit aquatic hosts whereas the remaining Anaplasmataceae are found in terrestrial hosts. Additionally, our evolutionary timeline places the directly transmitted marine Ca.Xc as the basal Anaplasmataceae, ancestral to both freshwater and terrestrial species with adaptations leading to more complex life cycles involving intermediate vectors or reservoir species; this supports the hypothesis of a marine origin for this bacterial family.}, } @article {pmid27893736, year = {2016}, author = {Caragata, EP and Rezende, FO and Simões, TC and Moreira, LA}, title = {Diet-Induced Nutritional Stress and Pathogen Interference in Wolbachia-Infected Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {11}, pages = {e0005158}, pmid = {27893736}, issn = {1935-2735}, mesh = {Aedes/*microbiology/parasitology/*physiology/virology ; Animals ; Dengue Virus/physiology ; Feeding Behavior ; Female ; Insect Vectors/*microbiology/parasitology/*physiology/virology ; Male ; Pest Control, Biological ; Plasmodium gallinaceum/physiology ; Stress, Physiological ; Wolbachia/*physiology ; }, abstract = {The pathogen interference phenotype greatly restricts infection with dengue virus (DENV) and other pathogens in Wolbachia-infected Aedes aegypti, and is a vital component of Wolbachia-based mosquito control. Critically, the phenotype's causal mechanism is complex and poorly understood, with recent evidence suggesting that the cause may be species specific. To better understand this important phenotype, we investigated the role of diet-induced nutritional stress on interference against DENV and the avian malarial parasite Plasmodium gallinaceum in Wolbachia-infected Ae. aegypti, and on physiological processes linked to the phenotype. Wolbachia-infected mosquitoes were fed one of four different concentrations of sucrose, and then challenged with either P. gallinaceum or DENV. Interference against P. gallinaceum was significantly weakened by the change in diet however there was no effect on DENV interference. Immune gene expression and H2O2 levels have previously been linked to pathogen interference. These traits were assayed for mosquitoes on each diet using RT-qPCR and the Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit, and it was observed that the change in diet did not significantly affect immune expression, but low carbohydrate levels led to a loss of ROS induction in Wolbachia-infected mosquitoes. Our data suggest that host nutrition may not influence DENV interference for Wolbachia-infected mosquitoes, but Plasmodium interference may be linked to both nutrition and oxidative stress. This pathogen-specific response to nutritional change highlights the complex nature of interactions between Wolbachia and pathogens in mosquitoes.}, } @article {pmid27881553, year = {2016}, author = {Bennuru, S and Cotton, JA and Ribeiro, JM and Grote, A and Harsha, B and Holroyd, N and Mhashilkar, A and Molina, DM and Randall, AZ and Shandling, AD and Unnasch, TR and Ghedin, E and Berriman, M and Lustigman, S and Nutman, TB}, title = {Stage-Specific Transcriptome and Proteome Analyses of the Filarial Parasite Onchocerca volvulus and Its Wolbachia Endosymbiont.}, journal = {mBio}, volume = {7}, number = {6}, pages = {}, pmid = {27881553}, issn = {2150-7511}, support = {ZIA AI000512/AI/NIAID NIH HHS/United States ; R21 AI126466/AI/NIAID NIH HHS/United States ; R24 AG042328/AG/NIA NIH HHS/United States ; T32 AI007180/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; 098051/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Onchocerca volvulus/chemistry/*genetics/*growth & development ; *Proteome ; *Symbiosis ; *Transcriptome ; Wolbachia/chemistry/*genetics/*growth & development ; }, abstract = {UNLABELLED: Onchocerciasis (river blindness) is a neglected tropical disease that has been successfully targeted by mass drug treatment programs in the Americas and small parts of Africa. Achieving the long-term goal of elimination of onchocerciasis, however, requires additional tools, including drugs, vaccines, and biomarkers of infection. Here, we describe the transcriptome and proteome profiles of the major vector and the human host stages (L1, L2, L3, molting L3, L4, adult male, and adult female) of Onchocerca volvulus along with the proteome of each parasitic stage and of its Wolbachia endosymbiont (wOv). In so doing, we have identified stage-specific pathways important to the parasite's adaptation to its human host during its early development. Further, we generated a protein array that, when screened with well-characterized human samples, identified novel diagnostic biomarkers of O. volvulus infection and new potential vaccine candidates. This immunomic approach not only demonstrates the power of this postgenomic discovery platform but also provides additional tools for onchocerciasis control programs.

IMPORTANCE: The global onchocerciasis (river blindness) elimination program will have to rely on the development of new tools (drugs, vaccines, biomarkers) to achieve its goals by 2025. As an adjunct to the completed genomic sequencing of O. volvulus, we used a comprehensive proteomic and transcriptomic profiling strategy to gain a comprehensive understanding of both the vector-derived and human host-derived parasite stages. In so doing, we have identified proteins and pathways that enable novel drug targeting studies and the discovery of novel vaccine candidates, as well as useful biomarkers of active infection.}, } @article {pmid27871813, year = {2017}, author = {Ye, YH and Seleznev, A and Flores, HA and Woolfit, M and McGraw, EA}, title = {Gut microbiota in Drosophila melanogaster interacts with Wolbachia but does not contribute to Wolbachia-mediated antiviral protection.}, journal = {Journal of invertebrate pathology}, volume = {143}, number = {}, pages = {18-25}, doi = {10.1016/j.jip.2016.11.011}, pmid = {27871813}, issn = {1096-0805}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Drosophila melanogaster/*microbiology ; *Gastrointestinal Microbiome/drug effects ; *Host-Pathogen Interactions ; Insect Viruses ; RNA Viruses ; Virus Diseases/*microbiology ; *Wolbachia ; }, abstract = {Animals experience near constant infection with microorganisms. A significant proportion of these microbiota reside in the alimentary tract. There is a growing appreciation for the roles gut microbiota play in host biology. The gut microbiota of insects, for example, have been shown to help the host overcome pathogen infection either through direct competition or indirectly by stimulating host immunity. These defenses may also be supplemented by coinfecting maternally inherited microbes such as Wolbachia. The presence of Wolbachia in a host can delay and/or reduce death caused by RNA viruses. Whether the gut microbiota of the host interacts with Wolbachia, or vice versa, the precise role of Wolbachia in antiviral protection is not known. In this study, we used 16S rDNA sequencing to characterise changes in gut microbiota composition in Drosophila melanogaster associated with Wolbachia infection and antibiotic treatment. We subsequently tested whether changes in gut composition via antibiotic treatment altered Wolbachia-mediated antiviral properties. We found that both antibiotics and Wolbachia significantly reduced the biodiversity of the gut microbiota without changing the total microbial load. We also showed that changing the gut microbiota composition with antibiotic treatment enhanced Wolbachia density but did not confer greater antiviral protection against Drosophila C virus to the host. We concluded there are significant interactions between Wolbachia and gut microbiota, but changing gut microbiota composition is not likely to be a means through which Wolbachia conveys antiviral protection to its host.}, } @article {pmid27869792, year = {2016}, author = {Choi, YJ and Tyagi, R and McNulty, SN and Rosa, BA and Ozersky, P and Martin, J and Hallsworth-Pepin, K and Unnasch, TR and Norice, CT and Nutman, TB and Weil, GJ and Fischer, PU and Mitreva, M}, title = {Genomic diversity in Onchocerca volvulus and its Wolbachia endosymbiont.}, journal = {Nature microbiology}, volume = {2}, number = {}, pages = {16207}, pmid = {27869792}, issn = {2058-5276}, support = {//Wellcome Trust/United Kingdom ; R01 AI081803/AI/NIAID NIH HHS/United States ; R01 GM097435/GM/NIGMS NIH HHS/United States ; U54 HG003079/HG/NHGRI NIH HHS/United States ; }, mesh = {Africa, Western ; Animals ; Ecuador ; Gene Flow ; *Genetic Variation ; Genotype ; Onchocerca volvulus/*classification/*genetics/isolation & purification/microbiology ; Phylogeography ; Uganda ; Wolbachia/*classification/*genetics ; }, abstract = {Ongoing elimination efforts have altered the global distribution of Onchocerca volvulus, the agent of river blindness, and further population restructuring is expected as efforts continue. Therefore, a better understanding of population genetic processes and their effect on biogeography is needed to support elimination goals. We describe O. volvulus genome variation in 27 isolates from the early 1990s (before widespread mass treatment) from four distinct locales: Ecuador, Uganda, the West African forest and the West African savanna. We observed genetic substructuring between Ecuador and West Africa and between the West African forest and savanna bioclimes, with evidence of unidirectional gene flow from savanna to forest strains. We identified forest:savanna-discriminatory genomic regions and report a set of ancestry informative loci that can be used to differentiate between forest, savanna and admixed isolates, which has not previously been possible. We observed mito-nuclear discordance possibly stemming from incomplete lineage sorting. The catalogue of the nuclear, mitochondrial and endosymbiont DNA variants generated in this study will support future basic and translational onchocerciasis research, with particular relevance for ongoing control programmes, and boost efforts to characterize drug, vaccine and diagnostic targets.}, } @article {pmid27869790, year = {2016}, author = {Cotton, JA and Bennuru, S and Grote, A and Harsha, B and Tracey, A and Beech, R and Doyle, SR and Dunn, M and Hotopp, JC and Holroyd, N and Kikuchi, T and Lambert, O and Mhashilkar, A and Mutowo, P and Nursimulu, N and Ribeiro, JM and Rogers, MB and Stanley, E and Swapna, LS and Tsai, IJ and Unnasch, TR and Voronin, D and Parkinson, J and Nutman, TB and Ghedin, E and Berriman, M and Lustigman, S}, title = {The genome of Onchocerca volvulus, agent of river blindness.}, journal = {Nature microbiology}, volume = {2}, number = {}, pages = {16216}, pmid = {27869790}, issn = {2058-5276}, support = {DP2 OD007372/OD/NIH HHS/United States ; R01 AI078314/AI/NIAID NIH HHS/United States ; T32 AI007180/AI/NIAID NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Genome, Bacterial ; *Genome, Helminth ; Onchocerca volvulus/*genetics ; Onchocerciasis, Ocular/*parasitology ; Wolbachia/genetics ; }, abstract = {Human onchocerciasis is a serious neglected tropical disease caused by the filarial nematode Onchocerca volvulus that can lead to blindness and chronic disability. Control of the disease relies largely on mass administration of a single drug, and the development of new drugs and vaccines depends on a better knowledge of parasite biology. Here, we describe the chromosomes of O. volvulus and its Wolbachia endosymbiont. We provide the highest-quality sequence assembly for any parasitic nematode to date, giving a glimpse into the evolution of filarial parasite chromosomes and proteomes. This resource was used to investigate gene families with key functions that could be potentially exploited as targets for future drugs. Using metabolic reconstruction of the nematode and its endosymbiont, we identified enzymes that are likely to be essential for O. volvulus viability. In addition, we have generated a list of proteins that could be targeted by Federal-Drug-Agency-approved but repurposed drugs, providing starting points for anti-onchocerciasis drug development.}, } @article {pmid27863487, year = {2016}, author = {Cheke, RA}, title = {Analyses of density-dependent effects are needed to understand how and when Wolbachia can control dengue vectors.}, journal = {BMC biology}, volume = {14}, number = {1}, pages = {99}, pmid = {27863487}, issn = {1741-7007}, mesh = {Aedes ; Animals ; Dengue ; *Dengue Virus ; Humans ; Insect Vectors ; Pest Control, Biological ; *Wolbachia ; }, abstract = {Releases of Wolbachia-infected mosquitoes have been shown to be an effective method of controlling Aedes aegypti, the main vector of dengue fever, in Australia. A study in BMC Biology from Penelope Hancock and others shows that incorporation of density-dependent effects into population models can provide major improvements in understanding how and when the infected populations can become established.See research article: https://bmcbiol.biomedcentral.com/articles/10.1186/s12915-016-0319-5 .}, } @article {pmid27863033, year = {2017}, author = {Hawlitschek, O and Morinière, J and Lehmann, GUC and Lehmann, AW and Kropf, M and Dunz, A and Glaw, F and Detcharoen, M and Schmidt, S and Hausmann, A and Szucsich, NU and Caetano-Wyler, SA and Haszprunar, G}, title = {DNA barcoding of crickets, katydids and grasshoppers (Orthoptera) from Central Europe with focus on Austria, Germany and Switzerland.}, journal = {Molecular ecology resources}, volume = {17}, number = {5}, pages = {1037-1053}, doi = {10.1111/1755-0998.12638}, pmid = {27863033}, issn = {1755-0998}, mesh = {Animals ; Austria ; *DNA Barcoding, Taxonomic ; DNA, Bacterial/genetics ; Electron Transport Complex IV/genetics ; Germany ; International Cooperation ; Orthoptera/*classification/*genetics ; Switzerland ; Wolbachia/genetics ; }, abstract = {We present a DNA barcoding study on the insect order Orthoptera that was generated in collaboration between four barcoding projects in three countries, viz. Barcoding Fauna Bavarica (Germany), German Barcode of Life, Austrian Barcode of Life and Swiss Barcode of Life. Our data set includes 748 COI sequences from 127 of the 162 taxa (78.4%) recorded in the three countries involved. Ninety-three of these 122 species (76.2%, including all Ensifera) can be reliably identified using DNA barcodes. The remaining 26 caeliferan species (families Acrididae and Tetrigidae) form ten clusters that share barcodes among up to five species, in three cases even across different genera, and in six cases even sharing individual barcodes. We discuss incomplete lineage sorting and hybridization as most likely causes of this phenomenon, as the species concerned are phylogenetically young and hybridization has been previously observed. We also highlight the problem of nuclear mitochondrial pseudogenes (numts), a known problem in the barcoding of orthopteran species, and the possibility of Wolbachia infections. Finally, we discuss the possible taxonomic implications of our barcoding results and point out future research directions.}, } @article {pmid27862998, year = {2017}, author = {Adam, N and Erler, T and Kallenbach, M and Kaltenpoth, M and Kunert, G and Baldwin, IT and Schuman, MC}, title = {Sex ratio of mirid populations shifts in response to hostplant co-infestation or altered cytokinin signaling .}, journal = {Journal of integrative plant biology}, volume = {59}, number = {1}, pages = {44-59}, pmid = {27862998}, issn = {1744-7909}, support = {293926/ERC_/European Research Council/International ; }, mesh = {Animals ; Cytokinins/*metabolism ; Female ; Hemiptera/microbiology/*physiology ; Host-Parasite Interactions ; Male ; Nutritional Physiological Phenomena ; Oviposition ; Plant Diseases/*parasitology ; Population Dynamics ; Reproduction ; *Sex Ratio ; *Signal Transduction ; Symbiosis ; Nicotiana/*parasitology ; Wolbachia/physiology ; }, abstract = {Herbivore species sharing a host plant often compete. In this study, we show that host plant-mediated interaction between two insect herbivores - a generalist and a specialist - results in a sex ratio shift of the specialist's offspring. We studied demographic parameters of the specialist Tupiocoris notatus (Hemiptera: Miridae) when co-infesting the host plant Nicotiana attenuata (Solanaceae) with the generalist leafhopper Empoasca sp. (Hemiptera: Cicadellidae). We show that the usually female-biased sex ratio of T. notatus shifts toward a higher male proportion in the offspring on plants co-infested by Empoasca sp. This sex ratio change did not occur after oviposition, nor is it due differential mortality of female and male nymphs. Based on pyrosequencing and PCR of bacterial 16S rRNA amplicons, we concluded that sex ratio shifts were unlikely to be due to infection with Wolbachia or other known sex ratio-distorting endosymbionts. Finally, we used transgenic lines of N. attenuata to evaluate if the sex ratio shift could be mediated by changes in general or specialized host plant metabolites. We found that the sex ratio shift occurred on plants deficient in two cytokinin receptors (irCHK2/3). Thus, cytokinin-regulated traits can alter the offspring sex ratio of the specialist T. notatus.}, } @article {pmid27859964, year = {2017}, author = {Pontieri, L and Schmidt, AM and Singh, R and Pedersen, JS and Linksvayer, TA}, title = {Artificial selection on ant female caste ratio uncovers a link between female-biased sex ratios and infection by Wolbachia endosymbionts.}, journal = {Journal of evolutionary biology}, volume = {30}, number = {2}, pages = {225-234}, doi = {10.1111/jeb.13012}, pmid = {27859964}, issn = {1420-9101}, mesh = {Animals ; Female ; *Ants/parasitology ; *Sex Ratio ; *Symbiosis ; *Wolbachia/pathogenicity ; }, abstract = {Social insect sex and caste ratios are well-studied targets of evolutionary conflicts, but the heritable factors affecting these traits remain unknown. To elucidate these factors, we carried out a short-term artificial selection study on female caste ratio in the ant Monomorium pharaonis. Across three generations of bidirectional selection, we observed no response for caste ratio, but sex ratios rapidly became more female-biased in the two replicate high selection lines and less female-biased in the two replicate low selection lines. We hypothesized that this rapid divergence for sex ratio was caused by changes in the frequency of infection by the heritable bacterial endosymbiont Wolbachia, because the initial breeding stock varied for Wolbachia infection, and Wolbachia is known to cause female-biased sex ratios in other insects. Consistent with this hypothesis, the proportions of Wolbachia-infected colonies in the selection lines changed rapidly, mirroring the sex ratio changes. Moreover, the estimated effect of Wolbachia on sex ratio (~13% female bias) was similar in colonies before and during artificial selection, indicating that this Wolbachia effect is likely independent of the effects of artificial selection on other heritable factors. Our study provides evidence for the first case of endosymbiont sex ratio manipulation in a social insect.}, } @article {pmid27855218, year = {2016}, author = {Amuzu, HE and McGraw, EA}, title = {Wolbachia-Based Dengue Virus Inhibition Is Not Tissue-Specific in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {11}, pages = {e0005145}, pmid = {27855218}, issn = {1935-2735}, mesh = {Aedes/immunology/*microbiology/*virology ; Animals ; Antibiosis ; Dengue/prevention & control/virology ; Dengue Virus/*physiology ; Fat Body/microbiology/virology ; Malpighian Tubules/microbiology/virology ; Mosquito Vectors/microbiology/virology ; Organ Specificity ; Salivary Glands/microbiology/virology ; *Symbiosis ; Viral Load ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Dengue fever, caused by the dengue virus (DENV), is now the most common arbovirus transmitted disease globally. One novel approach to control DENV is to use the endosymbiotic bacterium, Wolbachia pipientis, to limit DENV replication inside the primary mosquito vector, Aedes aegypti. Wolbachia that is naturally present in a range of insects reduces the capacity for viruses, bacteria, parasites and fungi to replicate inside insects. Wolbachia's mode of action is not well understood but may involve components of immune activation or competition with pathogens for limited host resources. The strength of Wolbachia-based anti DENV effects appear to correlate with bacterial density in the whole insect and in cell culture. Here we aimed to determine whether particular tissues, especially those with high Wolbachia densities or immune activity, play a greater role in mediating the anti DENV effect.

METHODOLOGY/FINDINGS: Ae. aegypti mosquito lines with and without Wolbachia (Wildtype) were orally fed DENV 3 and their viral loads subsequently measured over two time points post infection in the midgut, head, salivary glands, Malpighian tubules, fat body and carcass. We did not find correlations between Wolbachia densities and DENV loads in any tissue, nor with DENV loads in salivary glands, the endpoint of infection. This is in contrast with strong positive correlations between DENV loads in a range of tissues and salivary gland loads for Wildtype mosquitoes. Lastly, there was no evidence of a heightened role for tissues with known immune function including the fat body and the Malpighian tubules in Wolbachia's limitation of DENV.

CONCLUSION/SIGNIFICANCE: We conclude that the efficacy of DENV blocking in Wolbachia infected mosquitoes is not reliant on any particular tissue. This work therefore suggests that the mechanism of Wolbachia-based antiviral effects is either systemic or acts locally via processes that are fundamental to diverse cell types. We further conclude that the relationship between DENV blocking and Wolbachia density is not linear in mosquito tissues.}, } @article {pmid27849636, year = {2017}, author = {Kamtchum-Tatuene, J and Makepeace, BL and Benjamin, L and Baylis, M and Solomon, T}, title = {The potential role of Wolbachia in controlling the transmission of emerging human arboviral infections.}, journal = {Current opinion in infectious diseases}, volume = {30}, number = {1}, pages = {108-116}, pmid = {27849636}, issn = {1473-6527}, support = {IS-HPU-1112-10117/DH_/Department of Health/United Kingdom ; MC_PC_15090/MRC_/Medical Research Council/United Kingdom ; RP-PG-0108-10048/DH_/Department of Health/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/virology ; Animals ; Arbovirus Infections/*prevention & control/transmission/virology ; Dengue/prevention & control/transmission ; Dengue Virus/*physiology ; Humans ; Insect Vectors/*microbiology/virology ; *Virus Replication ; Wolbachia/*physiology ; }, abstract = {PURPOSE OF REVIEW: Wolbachia is a genus of Gram-negative intracellular bacteria that is naturally found in more than half of all arthropod species. These bacteria cannot only reduce the fitness and the reproductive capacities of arthropod vectors, but also increase their resistance to arthropod-borne viruses (arboviruses). This article reviews the evidence supporting a Wolbachia-based strategy for controlling the transmission of dengue and other arboviral infections.

RECENT FINDINGS: Studies conducted 1 year after the field release of Wolbachia-infected mosquitoes in Australia have demonstrated the suppression of dengue virus (DENV) replication in and dissemination by mosquitoes. Recent mathematical models show that this strategy could reduce the transmission of DENV by 70%. Consequently, the WHO is encouraging countries to boost the development and implementation of Wolbachia-based prevention strategies against other arboviral infections. However, the evidence regarding the efficacy of Wolbachia to prevent the transmission of other arboviral infections is still limited to an experimental framework with conflicting results in some cases. There is a need to demonstrate the efficacy of such strategies in the field under various climatic conditions, to select the Wolbachia strain that has the best pathogen interference/spread trade-off, and to continue to build community acceptance.

SUMMARY: Wolbachia represents a promising tool for controlling the transmission of arboviral infections that needs to be developed further. Long-term environmental monitoring will be necessary for timely detection of potential changes in Wolbachia/vector/virus interactions.}, } @article {pmid27843186, year = {2016}, author = {Mustafa, MS and Rastogi, V and Gupta, RK and Jain, S and Singh, PM and Gupta, A}, title = {Wolbachia: The selfish Trojan Horse in dengue control.}, journal = {Medical journal, Armed Forces India}, volume = {72}, number = {4}, pages = {373-376}, pmid = {27843186}, issn = {0377-1237}, abstract = {Dengue fever has re-emerged as a major public health challenge. Of late, several promising attempts have been made to control the disease with limited success. An innovative method of biological control of dengue is the use of the bacterium Wolbachia. Selected strains of Wolbachia have been introduced into Aedes aegypti to prevent transmission of dengue viruses by the vector. Wolbachia prevents dengue transmission by either directly blocking the virus or by decreasing the lifespan of the vector. The mechanism by which it causes these effects is not clearly understood. The main concern of this technique is the emergence of a new dengue virus serotype which may evade the protection offered by Wolbachia. The technique is environment friendly and holds promise for control of other vector borne diseases.}, } @article {pmid27834855, year = {2016}, author = {Dietzgen, RG and Mann, KS and Johnson, KN}, title = {Plant Virus-Insect Vector Interactions: Current and Potential Future Research Directions.}, journal = {Viruses}, volume = {8}, number = {11}, pages = {}, pmid = {27834855}, issn = {1999-4915}, mesh = {Animals ; Disease Transmission, Infectious ; Entomology/trends ; *Host-Pathogen Interactions ; Insect Vectors/*physiology/*virology ; Plant Diseases/virology ; Plant Viruses/*physiology ; Virology/trends ; }, abstract = {Acquisition and transmission by an insect vector is central to the infection cycle of the majority of plant pathogenic viruses. Plant viruses can interact with their insect host in a variety of ways including both non-persistent and circulative transmission; in some cases, the latter involves virus replication in cells of the insect host. Replicating viruses can also elicit both innate and specific defense responses in the insect host. A consistent feature is that the interaction of the virus with its insect host/vector requires specific molecular interactions between virus and host, commonly via proteins. Understanding the interactions between plant viruses and their insect host can underpin approaches to protect plants from infection by interfering with virus uptake and transmission. Here, we provide a perspective focused on identifying novel approaches and research directions to facilitate control of plant viruses by better understanding and targeting virus-insect molecular interactions. We also draw parallels with molecular interactions in insect vectors of animal viruses, and consider technical advances for their control that may be more broadly applicable to plant virus vectors.}, } @article {pmid27831765, year = {2016}, author = {Caragata, EP and Dutra, HL and O'Neill, SL and Moreira, LA}, title = {Zika control through the bacterium Wolbachia pipientis.}, journal = {Future microbiology}, volume = {11}, number = {}, pages = {1499-1502}, doi = {10.2217/fmb-2016-0177}, pmid = {27831765}, issn = {1746-0921}, mesh = {Aedes/*microbiology ; Animals ; Humans ; Insect Vectors ; *Mosquito Control ; Wolbachia/*physiology ; Zika Virus Infection/*prevention & control/transmission ; }, } @article {pmid27827425, year = {2016}, author = {Asad, S and Hall-Mendelin, S and Asgari, S}, title = {Downregulation of Aedes aegypti chromodomain helicase DNA binding protein 7/Kismet by Wolbachia and its effect on dengue virus replication.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {36850}, pmid = {27827425}, issn = {2045-2322}, mesh = {Aedes/enzymology/*microbiology/virology ; Animals ; Cell Line ; DNA Helicases/*genetics ; Dengue Virus/*physiology ; *Down-Regulation ; Female ; Gene Silencing ; Insect Proteins/genetics ; Sex Factors ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Dengue virus (DENV) is a mosquito-transmitted virus imposing a significant burden on human health around the world. Since current control strategies are not sufficient, there is an urgent need to find alternative methods to control DENV transmission. It has been demonstrated that introduction of Wolbachia pipientis in Aedes aegypti mosquitoes can impede DENV transmission with the mechanism(s) not fully understood. Recently, a number of studies have found the involvement of chromodomain DNA binding helicases in case of Human Immunodeficiency virus (HIV) and Influenza A virus infection. In this study, we have identified three chromodomain helicase DNA binding protein (CHD) genes in Ae. aegypti and looked at their response in the case of Wolbachia and DENV infections. Foremost amongst them we have found that AeCHD7/Kismet is significantly downregulated in the presence of Wolbachia infection only in female mosquitoes. Furthermore, AeCHD7 levels showed significant increase during DENV infection, and AeCHD7 depletion led to severe reduction in the replication of DENV. Our data have identified AeCHD7 as a novel Ae. aegypti host factor that is important for DENV replication, and Wolbachia downregulates it, which may contribute towards the mechanism(s) of limiting DENV replication.}, } @article {pmid27825343, year = {2016}, author = {Hancock, PA and White, VL and Ritchie, SA and Hoffmann, AA and Godfray, HC}, title = {Predicting Wolbachia invasion dynamics in Aedes aegypti populations using models of density-dependent demographic traits.}, journal = {BMC biology}, volume = {14}, number = {1}, pages = {96}, pmid = {27825343}, issn = {1741-7007}, mesh = {Aedes/*microbiology ; Animals ; Bayes Theorem ; *Models, Theoretical ; Wolbachia/*pathogenicity ; Zika Virus/pathogenicity ; }, abstract = {BACKGROUND: Arbovirus transmission by the mosquito Aedes aegypti can be reduced by the introduction and establishment of the endosymbiotic bacteria Wolbachia in wild populations of the vector. Wolbachia spreads by increasing the fitness of its hosts relative to uninfected mosquitoes. However, mosquito fitness is also strongly affected by population size through density-dependent competition for limited food resources. We do not understand how this natural variation in fitness affects symbiont spread, which limits our ability to design successful control strategies.

RESULTS: We develop a mathematical model to predict A. aegypti-Wolbachia dynamics that incorporates larval density-dependent variation in important fitness components of infected and uninfected mosquitoes. Our model explains detailed features of the mosquito-Wolbachia dynamics observed in two independent experimental A. aegypti populations, allowing the combined effects on dynamics of multiple density-dependent fitness components to be characterized. We apply our model to investigate Wolbachia field release dynamics, and show how invasion outcomes can depend strongly on the severity of density-dependent competition at the release site. Specifically, the ratio of released relative to wild mosquitoes required to attain a target infection frequency (at the end of a release program) can vary by nearly an order of magnitude. The time taken for Wolbachia to become established following releases can differ by over 2 years. These effects depend on the relative fitness of field and insectary-reared mosquitoes.

CONCLUSIONS: Models of Wolbachia invasion incorporating density-dependent demographic variation in the host population explain observed dynamics in experimental A. aegypti populations. These models predict strong effects of density-dependence on Wolbachia dynamics in field populations, and can assist in the effective use of Wolbachia to control the transmission of arboviruses such as dengue, chikungunya and zika.}, } @article {pmid27821433, year = {2017}, author = {Cooper, BS and Ginsberg, PS and Turelli, M and Matute, DR}, title = {Wolbachia in the Drosophila yakuba Complex: Pervasive Frequency Variation and Weak Cytoplasmic Incompatibility, but No Apparent Effect on Reproductive Isolation.}, journal = {Genetics}, volume = {205}, number = {1}, pages = {333-351}, pmid = {27821433}, issn = {1943-2631}, support = {F32 AI114176/AI/NIAID NIH HHS/United States ; R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Africa, Western ; Animals ; Crosses, Genetic ; Cytoplasm/metabolism/microbiology ; Drosophila melanogaster/genetics/metabolism/*microbiology ; Female ; Host-Pathogen Interactions ; Hybridization, Genetic/*genetics ; Male ; Reproduction/genetics ; Reproductive Isolation ; Sexual Behavior, Animal/physiology ; Species Specificity ; Wolbachia/genetics/*physiology ; }, abstract = {Three hybridizing species-the clade [(Drosophila yakuba, D. santomea), D. teissieri]-comprise the yakuba complex in the D. melanogaster subgroup. Their ranges overlap on Bioko and São Tomé, islands off west Africa. All three species are infected with Wolbachia-maternally inherited, endosymbiotic bacteria, best known for manipulating host reproduction to favor infected females. Previous analyses reported no cytoplasmic incompatibility (CI) in these species. However, we discovered that Wolbachia from each species cause intraspecific and interspecific CI. In D teissieri, analyses of F1 and backcross genotypes show that both host genotype and Wolbachia variation modulate CI intensity. Wolbachia-infected females seem largely protected from intraspecific and interspecific CI, irrespective of Wolbachia and host genotypes. Wolbachia do not affect host mating behavior or female fecundity, within or between species. The latter suggests little apparent effect of Wolbachia on premating or gametic reproductive isolation (RI) between host species. In nature, Wolbachia frequencies varied spatially for D. yakuba in 2009, with 76% (N = 155) infected on São Tomé, and only 3% (N = 36) infected on Bioko; frequencies also varied temporally in D. yakuba and D. santomea on São Tomé between 2009 and 2015. These temporal frequency fluctuations could generate asymmetries in interspecific mating success, and contribute to postzygotic RI. However, the fluctuations in Wolbachia frequencies that we observe also suggest that asymmetries are unlikely to persist. Finally, we address theoretical questions that our empirical findings raise about Wolbachia persistence when conditions fluctuate, and about the stable coexistence of Wolbachia and host variants that modulate Wolbachia effects.}, } @article {pmid27819272, year = {2016}, author = {Li, K and Chen, H and Jiang, J and Li, X and Xu, J and Ma, Y}, title = {Diversity of bacteriome associated with Phlebotomus chinensis (Diptera: Psychodidae) sand flies in two wild populations from China.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {36406}, pmid = {27819272}, issn = {2045-2322}, support = {SC1 AI112786/AI/NIAID NIH HHS/United States ; SC1 GM109326/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Biodiversity ; Ecosystem ; Enterobacteriaceae/classification/genetics/isolation & purification ; *Microbiota ; Phylogeny ; Principal Component Analysis ; Pseudomonas/classification/genetics/isolation & purification ; Psychodidae/*microbiology ; RNA, Ribosomal, 16S/isolation & purification/metabolism ; Rickettsia/classification/genetics/isolation & purification ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/isolation & purification ; }, abstract = {Sand fly Phlebotomus chinensis is a primary vector of transmission of visceral leishmaniasis in China. The sand flies have adapted to various ecological niches in distinct ecosystems. Characterization of the microbial structure and function will greatly facilitate the understanding of the sand fly ecology, which would provide critical information for developing intervention strategy for sand fly control. In this study we compared the bacterial composition between two populations of Ph. chinensis from Henan and Sichuan, China. The phylotypes were taxonomically assigned to 29 genera of 19 families in 9 classes of 5 phyla. The core bacteria include Pseudomonas and enterobacteria, both are shared in the sand flies in the two regions. Interestingly, the endosymbionts Wolbachia and Rickettsia were detected only in Henan, while the Rickettsiella and Diplorickettsia only in Sichuan. The intracellular bacteria Rickettsia, Rickettsiella and Diplorickettsia were reported for the first time in sand flies. The influence of sex and feeding status on the microbial structure was also detected in the two populations. The findings suggest that the ecological diversity of sand fly in Sichuan and Henan may contribute to shaping the structure of associated microbiota. The structural classification paves the way to function characterization of the sand fly associated microbiome.}, } @article {pmid27811722, year = {2016}, author = {Mock, A and Tajovský, K and Žurovcová, M and Jarošová, A and Kocourek, P and Gruber, J and Angyal, D and Spelda, J}, title = {Hungarosoma bokori Verhoeff, 1928 (Diplopoda: Chordeumatida): new insights into its taxonomy, systematics, molecular genetics, biogeography and ecology.}, journal = {Zootaxa}, volume = {4178}, number = {2}, pages = {234-256}, doi = {10.11646/zootaxa.4178.2.4}, pmid = {27811722}, issn = {1175-5334}, mesh = {Animals ; Arthropods/*anatomy & histology/*classification/genetics/microbiology ; Caves ; DNA Barcoding, Taxonomic ; Ecosystem ; Female ; Hungary ; Male ; Parthenogenesis ; Species Specificity ; Wolbachia/genetics ; }, abstract = {Hungarosoma bokori Verhoeff, 1928 is a millipede species which was originally classified solely on the basis of a female specimen. Subsequently, a long history of field searching for and surmising about the systematic position of this small, enigmatic species followed. In April 2013, 85 years after its first description, a series of nine specimens were sampled in the type locality, the Abaliget Cave, in southern Hungary. An adult male was collected for the first time, along with females and juveniles. Descriptions of the gonopods and the female vulvae, both important for considerations of the systematic position of the species, are presented for the first time. Revision and re-designation of the type material was made.The cryptic life of the species is connected with its activity in winter, and its known fragmented distribution corresponds with its presence in undisturbed microhabitats having a specific microclimate, often in the soil at cave entrances.Molecular methods showed a positive detection of the intracellular prokaryotic parasite Wolbachia in H. bokori, reflecting its highly probable parthenogenetic character in the main part of its known area of occurrence. This is the first demonstration of Wolbachia in a millipede.The legitimacy of the family Hungarosomatidae Ceuca, 1974, as a separate taxon was analysed using morphological and molecular approaches. Results of both methods confirmed the existence of a distinct phyletic line. DNA barcoding has shown its closest position to Attemsiidae Verhoeff, 1899, or Neoatractosomatidae Verhoeff, 1901. Based on records from Austria, the Czech Republic, Hungary and Slovakia, the residual circum-pannonic distribution that the whole genus (family) probably represents is proposed.}, } @article {pmid27808208, year = {2016}, author = {Callaway, E}, title = {Rio fights Zika with biggest release yet of bacteria-infected mosquitoes.}, journal = {Nature}, volume = {539}, number = {7627}, pages = {17-18}, doi = {10.1038/nature.2016.20878}, pmid = {27808208}, issn = {1476-4687}, mesh = {Aedes/*microbiology/*physiology/virology ; Animals ; Australia ; Brazil/epidemiology ; Chikungunya virus/growth & development ; China ; Cities/epidemiology ; Colombia ; Dengue/epidemiology/prevention & control/transmission/virology ; Female ; Fertility ; Humans ; Indonesia ; Male ; Polynesia ; Wolbachia/*pathogenicity ; Zika Virus/*growth & development ; Zika Virus Infection/epidemiology/*prevention & control/*transmission/virology ; }, } @article {pmid27806047, year = {2016}, author = {Rašić, G and Filipović, I and Callahan, AG and Stanford, D and Chan, A and Lam-Phua, SG and Tan, CH and Hoffmann, AA}, title = {The queenslandensis and the type Form of the Dengue Fever Mosquito (Aedes aegypti L.) Are Genomically Indistinguishable.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {11}, pages = {e0005096}, pmid = {27806047}, issn = {1935-2735}, mesh = {Aedes/classification/*genetics ; Animals ; Dengue/*transmission ; Female ; Genetic Variation ; Genomics ; Genotype ; Humans ; Insect Vectors/classification/*genetics ; Male ; Phylogeny ; Polymorphism, Single Nucleotide ; Queensland ; Singapore ; }, abstract = {BACKGROUND: The mosquito Aedes aegypti (L.) is a major vector of viral diseases like dengue fever, Zika and chikungunya. Aedes aegypti exhibits high morphological and behavioral variation, some of which is thought to be of epidemiological significance. Globally distributed domestic Ae. aegypti have often been grouped into (i) the very pale variety queenslandensis and (ii) the type form. Because the two color forms co-occur across most of their range, there is interest in understanding how freely they interbreed. This knowledge is particularly important for control strategies that rely on mating compatibilities between the release and target mosquitoes, such as Wolbachia releases and SIT. To address this question, we analyzed nuclear and mitochondrial genome-wide variation in the co-occurring pale and type Ae. aegypti from northern Queensland (Australia) and Singapore.

METHODS/FINDINGS: We typed 74 individuals at a 1170 bp-long mitochondrial sequence and at 16,569 nuclear SNPs using a customized double-digest RAD sequencing. 11/29 genotyped individuals from Singapore and 11/45 from Queensland were identified as var. queenslandensis based on the diagnostic scaling patterns. We found 24 different mitochondrial haplotypes, seven of which were shared between the two forms. Multivariate genetic clustering based on nuclear SNPs corresponded to individuals' geographic location, not their color. Several family groups consisted of both forms and three queenslandensis individuals were Wolbachia infected, indicating previous breeding with the type form which has been used to introduce Wolbachia into Ae. aegypti populations.

CONCLUSION: Aedes aegypti queenslandensis are genomically indistinguishable from the type form, which points to these forms freely interbreeding at least in Australia and Singapore. Based on our findings, it is unlikely that the presence of very pale Ae. aegypti will affect the success of Aedes control programs based on Wolbachia-infected, sterile or RIDL mosquitoes.}, } @article {pmid27802941, year = {2016}, author = {Mayor, S}, title = {Sixty seconds on . . . Wolbachia.}, journal = {BMJ (Clinical research ed.)}, volume = {355}, number = {}, pages = {i5847}, doi = {10.1136/bmj.i5847}, pmid = {27802941}, issn = {1756-1833}, mesh = {Aedes/*microbiology/virology ; Animals ; Chikungunya Fever/transmission ; Dengue/transmission ; Humans ; Insect Vectors/*microbiology/virology ; Virus Diseases/*transmission ; Virus Replication/physiology ; Wolbachia/*physiology ; Zika Virus Infection/transmission ; }, } @article {pmid27802919, year = {2017}, author = {Budachetri, K and Williams, J and Mukherjee, N and Sellers, M and Moore, F and Karim, S}, title = {The microbiome of neotropical ticks parasitizing on passerine migratory birds.}, journal = {Ticks and tick-borne diseases}, volume = {8}, number = {1}, pages = {170-173}, pmid = {27802919}, issn = {1877-9603}, support = {P20 GM103476/GM/NIGMS NIH HHS/United States ; P20 RR016476/RR/NCRR NIH HHS/United States ; }, mesh = {*Animal Migration ; Animals ; Bacteria/genetics/*isolation & purification ; Bird Diseases/*parasitology ; DNA, Bacterial/genetics ; *Microbiota ; Passeriformes/*parasitology ; Tick Infestations/parasitology/*veterinary ; }, abstract = {Seasonal migration of passerine birds between temperate North America and tropical Central and South America is an ecological phenomenon. Migration of birds has been associated with the introduction of ectoparasites like ticks or tick-borne pathogens across the avian migration routes. In this study, the microbial diversity was determined in the ticks and bird DNA samples using 454 pyrosequencing of bacterial 16S rRNA gene. Tick DNA samples showed the dominance of genera Lactococcus, Francisella, Raoultella, Wolbachia and Rickettsia across all the ticks, but birds DNA did not share common microbial diversity with ticks. Furthermore, "Candidatus Rickettsia amblyommii" infection in the 91 ticks collected off the songbirds was also quantified by qPCR assay. Interestingly, "Candidatus R. amblyommii" was tested positive in 24 ticks (26% infection), and infection varied from as low as three copies to thousands of copies, but bird blood samples showed no amplification. Our results provide evidence that songbirds serve as transport carrier for immature ticks, and less likely to be a reservoir for "Candidatus R. amblyommii".}, } @article {pmid27718295, year = {2016}, author = {Coon, KL and Brown, MR and Strand, MR}, title = {Mosquitoes host communities of bacteria that are essential for development but vary greatly between local habitats.}, journal = {Molecular ecology}, volume = {25}, number = {22}, pages = {5806-5826}, pmid = {27718295}, issn = {1365-294X}, support = {R01 AI106892/AI/NIAID NIH HHS/United States ; T32 GM007103/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Bacteria/*classification ; Ecosystem ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Southeastern United States ; }, abstract = {Mosquitoes are insects of interest because several species vector disease-causing pathogens to humans and other vertebrates. We previously reported that mosquitoes from long-term laboratory cultures require living bacteria in their gut to develop, but development does not depend on particular species of bacteria. Here, we focused on three distinct but interrelated areas of study to better understand the role of bacteria in mosquito development by studying field and laboratory populations of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus from the southeastern United States. Sequence analysis of bacterial 16S rRNA gene amplicons showed that bacterial community composition differed substantially in larvae from different collection sites, whereas larvae from the same site shared similarities. Although previously unknown to be infected by Wolbachia, results also indicated that Ae. aegypti from one field site hosted a dual infection. Regardless of collection site or factors like Wolbachia infection, however, each mosquito species required living bacteria in their digestive tract to develop. Results also identified several concerns in using antibiotics to eliminate the bacterial community in larvae in order to study its developmental consequences. Altogether, our results indicate that several mosquito species require living bacteria for development. We also hypothesize these species do not rely on particular bacteria because larvae do not reliably encounter the same bacteria in the aquatic habitats they develop in.}, } @article {pmid27784636, year = {2016}, author = {Puggioli, A and Calvitti, M and Moretti, R and Bellini, R}, title = {wPip Wolbachia contribution to Aedes albopictus SIT performance: Advantages under intensive rearing.}, journal = {Acta tropica}, volume = {164}, number = {}, pages = {473-481}, doi = {10.1016/j.actatropica.2016.10.014}, pmid = {27784636}, issn = {1873-6254}, mesh = {Aedes/*physiology ; Animals ; Female ; Infertility/physiopathology ; Male ; Mosquito Vectors/*physiology ; Pupa ; Reproduction/*drug effects ; Sexual Behavior, Animal/*drug effects ; Wolbachia/*physiology ; }, abstract = {As a part of a project aiming at the suppression of the mosquito vector Aedes albopictus, a specific Ae. albopictus line producing sterile males, ARwP, was tested for its suitability to intense rearing conditions compatible with mass production and field release. This line was developed by the Italian National Agency for New Technologies, Energy and Sustainable Economic Development thanks to the artificial infection with a heterologous Wolbachia strain, resulting in a bidirectional incompatibility pattern with wild-type Ae. albopictus. ARwP was reared under Standard Operating Procedures at the Centro Agricoltura Ambiente and compared with a wild-type strain in terms of time of pupation onset, production of male pupae in the following 24h and mechanical sexing efficacy. Mating competitiveness of ARwP males was also evaluated in comparison with irradiated wild-type males in large field enclosures. ARwP males demonstrated a significantly shorter time of pupation onset, a higher rate of production of male pupae in the following 24h and a lower percentage of residual contaminant females when applying mechanical sexing procedures. In addition, ARwP males were more efficient than wild-types in competing for wild-type females in large enclosures, thus inducing a level of sterility significantly higher than that expected for an equal mating competitiveness. These results encourage the use of this Ae. albopictus strain as suppression tool against Ae. albopictus based on considerations thoroughly discussed in the manuscript.}, } @article {pmid27770026, year = {2016}, author = {Lin, Y and Chen, ZX and Oliver, B and Harbison, ST}, title = {Microenvironmental Gene Expression Plasticity Among Individual Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {6}, number = {12}, pages = {4197-4210}, pmid = {27770026}, issn = {2160-1836}, mesh = {Animals ; Cluster Analysis ; Drosophila melanogaster/*genetics ; *Environment ; Female ; Gene Expression Profiling ; *Gene Expression Regulation ; *Gene-Environment Interaction ; Genetic Variation ; Genotype ; High-Throughput Nucleotide Sequencing ; Male ; Sex Characteristics ; Sex Factors ; Transcriptome ; }, abstract = {Differences in phenotype among genetically identical individuals exposed to the same environmental condition are often noted in genetic studies. Despite this commonplace observation, little is known about the causes of this variability, which has been termed microenvironmental plasticity. One possibility is that stochastic or technical sources of variance produce these differences. A second possibility is that this variation has a genetic component. We have explored gene expression robustness in the transcriptomes of 730 individual Drosophila melanogaster of 16 fixed genotypes, nine of which are infected with Wolbachia Three replicates of flies were grown, controlling for food, day/night cycles, humidity, temperature, sex, mating status, social exposure, and circadian timing of RNA extraction. Despite the use of inbred genotypes, and carefully controlled experimental conditions, thousands of genes were differentially expressed, revealing a unique and dynamic transcriptional signature for each individual fly. We found that 23% of the transcriptome was differentially expressed among individuals, and that the variability in gene expression among individuals is influenced by genotype. This transcriptional variation originated from specific gene pathways, suggesting a plastic response to the microenvironment; but there was also evidence of gene expression differences due to stochastic fluctuations. These observations reveal previously unappreciated genetic sources of variability in gene expression among individuals, which has implications for complex trait genetics and precision medicine.}, } @article {pmid27742209, year = {2016}, author = {Wu, CH and Zong, Q and Du, AL and Zhang, W and Yao, HC and Yu, XQ and Wang, YF}, title = {Knockdown of Dynamitin in testes significantly decreased male fertility in Drosophila melanogaster.}, journal = {Developmental biology}, volume = {420}, number = {1}, pages = {79-89}, doi = {10.1016/j.ydbio.2016.10.007}, pmid = {27742209}, issn = {1095-564X}, mesh = {Animals ; Cell Nucleus/metabolism ; Drosophila Proteins/*metabolism ; Drosophila melanogaster/*physiology ; Dyneins/metabolism ; Fertility ; Gene Expression Regulation ; *Gene Knockdown Techniques ; Intermediate Filament Proteins/*metabolism ; Male ; Mitochondria/metabolism ; Spermatids/metabolism ; Spermatogenesis ; Testis/*physiology ; Tubulin/metabolism ; }, abstract = {Dynamitin (Dmn) is a major component of dynactin, a multiprotein complex playing important roles in a variety of intracellular motile events. We previously found that Wolbachia bacterial infection resulted in a reduction of Dmn protein. As Wolbachia may modify sperm in male hosts, we speculate that Dmn may have a function in male fertility. Here we used nosGal4 to drive Dmn knock down in testes of Drosophila melanogaster to investigate the functions of Dmn in spermatogenesis. We found that knockdown of Dmn in testes dramatically decreased male fertility, overexpression of Dmn in Wolbachia-infected males significantly rescued male fertility, indicating an important role of Dmn in inducing male fertility defects following Wolbachia infection. Some scattered immature sperm with late canoe-shaped head distributed in the end of Dmn knockdown testis and only about half mature sperm were observed in the Dmn knockdown testis relative to those in the control. Transmission electron microscopy (TEM) exhibited fused spermatids in cysts and abnormal mitochondrial derivatives. Immunofluorescence staining showed significantly less abundance of tubulin around the nucleus of spermatid and scattered F-actin cones to different extents in the individualization complex (IC) during spermiogenesis in Dmn knockdown testes, which may disrupt the nuclear condensation and sperm individualization. Since dynein-dynactin complex has been shown to mediate transport of many cellular components, including mRNAs and organelles, these results suggest that Dmn may play an important role in Drosophila spermiogenesis by affecting transport of many important cytoplasmic materials.}, } @article {pmid27702765, year = {2016}, author = {Fromont, C and Riegler, M and Cook, JM}, title = {Phylogeographic analyses of bacterial endosymbionts in fig homotomids (Hemiptera: Psylloidea) reveal codiversification of both primary and secondary endosymbionts.}, journal = {FEMS microbiology ecology}, volume = {92}, number = {12}, pages = {}, doi = {10.1093/femsec/fiw205}, pmid = {27702765}, issn = {1574-6941}, mesh = {Animals ; Base Composition/genetics ; Biological Evolution ; DNA, Mitochondrial/genetics ; Enterobacteriaceae/classification/genetics/isolation & purification ; Evolution, Molecular ; Ficus ; Halomonadaceae/genetics/*isolation & purification ; Hemiptera/*microbiology ; Phylogeny ; Phylogeography ; Symbiosis/*genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {While obligate primary (P-) endosymbionts usually cospeciate with their insect hosts, less is known about codiversification of secondary (S-) endosymbionts that are generally considered facultative. Typically, insects of the superfamily Psylloidea harbour one P- (Carsonella) and at least one S-endosymbiont, thought to compensate for Carsonella genome reduction. Most codiversification studies have used phylogenies of psyllids and their endosymbionts across and within host families or genera, but few have explored patterns within species. We focussed on P- and S-endosymbionts of three Mycopsylla (Homotomidae) species to explore whether they have congruent phylogenies and within-species geographic structures. The P-endosymbiont Carsonella, a S-endosymbiont and Wolbachia all had 100% prevalence, while Arsenophonus was only found in one species at low prevalence. Congruent phylogenies of Mycopsylla and P-endosymbionts across populations and species support strict cospeciation. S-endosymbiont phylogenies were also congruent across host species but low genetic variation in the S-endosymbiont was not correlated with host phylogeography, possibly due to a shorter evolutionary association. Between species, Wolbachia and Mycopsylla phylogenies were incongruent, probably due to horizontal transmission events. Our study is the first to explore endosymbionts of Mycopsylla and further supports the codivergence of Psylloidea hosts and P-endosymbionts, with obligate host interactions for both P- and S-endosymbionts.}, } @article {pmid27768689, year = {2016}, author = {Sikulu-Lord, MT and Milali, MP and Henry, M and Wirtz, RA and Hugo, LE and Dowell, FE and Devine, GJ}, title = {Near-Infrared Spectroscopy, a Rapid Method for Predicting the Age of Male and Female Wild-Type and Wolbachia Infected Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {10}, pages = {e0005040}, pmid = {27768689}, issn = {1935-2735}, mesh = {Aedes/*growth & development/*microbiology ; Animals ; Female ; Insect Control ; Insect Vectors/*growth & development/*microbiology ; Male ; Pest Control, Biological ; Spectroscopy, Near-Infrared/*methods ; Wolbachia/*physiology ; }, abstract = {Estimating the age distribution of mosquito populations is crucial for assessing their capacity to transmit disease and for evaluating the efficacy of available vector control programs. This study reports on the capacity of the near-infrared spectroscopy (NIRS) technique to rapidly predict the ages of the principal dengue and Zika vector, Aedes aegypti. The age of wild-type males and females, and males and females infected with wMel and wMelPop strains of Wolbachia pipientis were characterized using this method. Calibrations were developed using spectra collected from their heads and thoraces using partial least squares (PLS) regression. A highly significant correlation was found between the true and predicted ages of mosquitoes. The coefficients of determination for wild-type females and males across all age groups were R2 = 0.84 and 0.78, respectively. The coefficients of determination for the age of wMel and wMelPop infected females were 0.71 and 0.80, respectively (P< 0.001 in both instances). The age of wild-type female Ae. aegypti could be identified as < or ≥ 8 days old with an accuracy of 91% (N = 501), whereas female Ae. aegypti infected with wMel and wMelPop were differentiated into the two age groups with an accuracy of 83% (N = 284) and 78% (N = 229), respectively. Our results also indicate NIRS can distinguish between young and old male wild-type, wMel and wMelPop infected Ae. aegypti with accuracies of 87% (N = 253), 83% (N = 277) and 78% (N = 234), respectively. We have demonstrated the potential of NIRS as a predictor of the age of female and male wild-type and Wolbachia infected Ae. aegypti mosquitoes under laboratory conditions. After field validation, the tool has the potential to offer a cheap and rapid alternative for surveillance of dengue and Zika vector control programs.}, } @article {pmid27756651, year = {2016}, author = {Wiwatanaratanabutr, I and Grandjean, F}, title = {Impacts of temperature and crowding on sex ratio, fecundity and Wolbachia infection intensity in the copepod, Mesocyclops thermocyclopoides.}, journal = {Journal of invertebrate pathology}, volume = {141}, number = {}, pages = {18-23}, doi = {10.1016/j.jip.2016.10.003}, pmid = {27756651}, issn = {1096-0805}, mesh = {Animals ; Copepoda/*microbiology ; Female ; Fertility ; Gram-Negative Bacterial Infections/*veterinary ; Hot Temperature ; Male ; Real-Time Polymerase Chain Reaction ; Sex Ratio ; *Wolbachia ; }, abstract = {Wolbachia are a group of intracellular bacteria that cause reproductive alterations in arthropods. Here, we describe the effects of two environmental factors (crowding and temperature) on phenotypic expression of feminization, the host's fecundity and Wolbachia infection intensity among life cycle stages in the naturally Wolbachia-infected copepod, Mesocyclops thermocyclopoides. The copepod was first found to be co-infected with Wolbachia A- and B-supergroups Wolbachia strains based on wsp primers. The relative Wolbachia infection intensity within individuals was determined using quantitative real-time PCR and was significantly higher in the B-supergroup than in the A-supergroup. Experimental results of temperature effect on bacterial density in each developmental stage revealed a significant decrease in Wolbachia infection intensity following exposure to high temperature (37°C) in both sexes and implied that Wolbachia might survive in room temperature (25°C) better than in high temperature. Experimental results of crowding effects on Wolbachia infection intensity suggested a negative correlation between copepod nauplii and Wolbachia infection intensity. No effect of rearing temperature on the sex ratio was reported although the fecundity was significantly decreased by high temperature. The results showed that Wolbachia infection intensity to be correlated with crowding conditions and was decreased following exposure of elevated temperature.}, } @article {pmid27760540, year = {2016}, author = {Jiggins, FM}, title = {Open questions: how does Wolbachia do what it does?.}, journal = {BMC biology}, volume = {14}, number = {1}, pages = {92}, pmid = {27760540}, issn = {1741-7007}, mesh = {Aedes/microbiology ; Animals ; Female ; *Host-Pathogen Interactions ; Lepidoptera/microbiology ; Male ; Sex Ratio ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {A common symbiont of insects, the bacterium Wolbachia has been implicated in phenomena as diverse as sex determination, pathogen defence and speciation and is being used in public health programs to prevent mosquitoes transmitting disease. Despite decades of research, we know remarkably little about how it exerts its effects.}, } @article {pmid27760142, year = {2016}, author = {Etebari, K and Asad, S and Zhang, G and Asgari, S}, title = {Identification of Aedes aegypti Long Intergenic Non-coding RNAs and Their Association with Wolbachia and Dengue Virus Infection.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {10}, pages = {e0005069}, pmid = {27760142}, issn = {1935-2735}, mesh = {Aedes/*genetics/microbiology/virology ; Animals ; Dengue Virus/*physiology ; Genome, Insect ; *Host-Pathogen Interactions/genetics ; Mosquito Vectors/*genetics/microbiology/virology ; Pest Control, Biological ; RNA Interference ; RNA, Long Noncoding/*genetics/isolation & purification/metabolism ; Transcriptome ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Long intergenic non-coding RNAs (lincRNAs) are appearing as an important class of regulatory RNAs with a variety of biological functions. The aim of this study was to identify the lincRNA profile in the dengue vector Aedes aegypti and evaluate their potential role in host-pathogen interaction. The majority of previous RNA-Seq transcriptome studies in Ae. aegypti have focused on the expression pattern of annotated protein coding genes under different biological conditions. Here, we used 35 publically available RNA-Seq datasets with relatively high depth to screen the Ae. aegypti genome for lincRNA discovery. This led to the identification of 3,482 putative lincRNAs. These lincRNA genes displayed a slightly lower GC content and shorter transcript lengths compared to protein-encoding genes. Ae. aegypti lincRNAs also demonstrate low evolutionary sequence conservation even among closely related species such as Culex quinquefasciatus and Anopheles gambiae. We examined their expression in dengue virus serotype 2 (DENV-2) and Wolbachia infected and non-infected adult mosquitoes and Aa20 cells. The results revealed that DENV-2 infection increased the abundance of a number of host lincRNAs, from which some suppress viral replication in mosquito cells. RNAi-mediated silencing of lincRNA_1317 led to enhancement in viral replication, which possibly indicates its potential involvement in the host anti-viral defense. A number of lincRNAs were also differentially expressed in Wolbachia-infected mosquitoes. The results will facilitate future studies to unravel the function of lncRNAs in insects and may prove to be beneficial in developing new ways to control vectors or inhibit replication of viruses in them.}, } @article {pmid27734242, year = {2016}, author = {Zhang, X and Tang, S and Cheke, RA and Zhu, H}, title = {Modeling the Effects of Augmentation Strategies on the Control of Dengue Fever With an Impulsive Differential Equation.}, journal = {Bulletin of mathematical biology}, volume = {78}, number = {10}, pages = {1968-2010}, doi = {10.1007/s11538-016-0208-7}, pmid = {27734242}, issn = {1522-9602}, support = {//CIHR/Canada ; }, mesh = {Aedes/microbiology/virology ; Animals ; *Biological Control Agents ; Computer Simulation ; Dengue/*prevention & control/transmission ; Female ; Humans ; Male ; Mathematical Concepts ; Models, Biological ; Mosquito Vectors/microbiology/virology ; Sex Ratio ; Wolbachia/physiology ; }, abstract = {Dengue fever has rapidly become the world's most common vector-borne viral disease. Use of endosymbiotic Wolbachia is an innovative technology to prevent vector mosquitoes from reproducing and so break the cycle of dengue transmission. However, strategies such as population eradication and replacement will only succeed if appropriate augmentations with Wolbachia-infected mosquitoes that take account of a variety of factors are carried out. Here, we describe the spread of Wolbachia in mosquito populations using an impulsive differential system with four state variables, incorporating the effects of cytoplasmic incompatibility and the augmentation of Wolbachia-infected mosquitoes with different sex ratios. We then evaluated (a) how each parameter value contributes to the success of population replacement; (b) how different release quantities of infected mosquitoes with different sex ratios affect the success of population suppression or replacement; and (c) how the success of these two strategies can be realized to block the transmission of dengue fever. Analysis of the system's stability, bifurcations and sensitivity reveals the existence of forward and backward bifurcations, multiple attractors and the contribution of each parameter to the success of the strategies. The results indicate that the initial density of mosquitoes, the quantities of mosquitoes released in augmentations and their sex ratios have impacts on whether or not the strategies of population suppression or replacement can be achieved. Therefore, successful strategies rely on selecting suitable strains of Wolbachia and carefully designing the mosquito augmentation program.}, } @article {pmid27752109, year = {2016}, author = {Tamarozzi, F and Turner, JD and Pionnier, N and Midgley, A and Guimaraes, AF and Johnston, KL and Edwards, SW and Taylor, MJ}, title = {Wolbachia endosymbionts induce neutrophil extracellular trap formation in human onchocerciasis.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {35559}, pmid = {27752109}, issn = {2045-2322}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Cell-Derived Microparticles/metabolism ; Doxycycline/pharmacology/therapeutic use ; Extracellular Traps/drug effects/*metabolism ; Female ; Fluorescent Antibody Technique ; Humans ; Lipopeptides/metabolism ; Mice ; Mice, Inbred C57BL ; Microspheres ; Neutrophil Infiltration/drug effects ; Neutrophils/drug effects/*metabolism ; Onchocerciasis/drug therapy/*microbiology/pathology ; *Symbiosis/drug effects ; Toll-Like Receptor 6/metabolism ; Wolbachia/drug effects/*physiology ; }, abstract = {The endosymbiotic bacteria, Wolbachia, induce neutrophilic responses to the human helminth pathogen Onchocerca volvulus. The formation of Neutrophil Extracellular Traps (NETs), has been implicated in anti-microbial defence, but has not been identified in human helminth infection. Here, we demonstrate NETs formation in human onchocerciasis. Extracellular NETs and neutrophils were visualised around O. volvulus in nodules excised from untreated patients but not in nodules from patients treated with the anti-Wolbachia drug, doxycycline. Whole Wolbachia or microspheres coated with a synthetic Wolbachia lipopeptide (WoLP) of the major nematode Wolbachia TLR2/6 ligand, peptidoglycan associated lipoprotein, induced NETosis in human neutrophils in vitro. TLR6 dependency of Wolbachia and WoLP NETosis was demonstrated using purified neutrophils from TLR6 deficient mice. Thus, we demonstrate for the first time that NETosis occurs during natural human helminth infection and demonstrate a mechanism of NETosis induction via Wolbachia endobacteria and direct ligation of Wolbachia lipoprotein by neutrophil TLR2/6.}, } @article {pmid27748992, year = {2017}, author = {Hunter, MS and Asiimwe, P and Himler, AG and Kelly, SE}, title = {Host nuclear genotype influences phenotype of a conditional mutualist symbiont.}, journal = {Journal of evolutionary biology}, volume = {30}, number = {1}, pages = {141-149}, doi = {10.1111/jeb.12993}, pmid = {27748992}, issn = {1420-9101}, mesh = {Animals ; Female ; Genetic Variation ; *Genotype ; *Hemiptera ; *Phenotype ; *Rickettsia ; *Symbiosis ; }, abstract = {Arthropods commonly carry maternally inherited intracellular bacterial symbionts that may profoundly influence host biology and evolution. The intracellular symbiont Rickettsia sp. nr. bellii swept rapidly into populations of the sweetpotato whitefly Bemisia tabaci in the south-western USA. Previous laboratory experiments showed female-bias and fitness benefits were associated with Rickettsia infection, potentially explaining the high frequencies of infection observed in field populations, but the effects varied with whitefly genetic line. Here, we explored whether host extranuclear or nuclear genes influenced the variation in the Rickettsia-host phenotype in two genetic lines of the whitefly host, each with Rickettsia-infected and uninfected sublines. Introgression between the Rickettsia-infected subline of one genetic line and the Rickettsia-uninfected subline of the other was used to create two new sublines, each with the maternally inherited extranuclear genetic lineages of one line (Rickettsia, two other symbionts and the mitochondria) and the nuclear genotype of the other. Performance assays comparing the original and new lines showed that in addition to Rickettsia, the interaction of Rickettsia infection with host nuclear genotype influenced development time and the sex ratio of the progeny, whereas the extranuclear genotype did not. Host nuclear genotype, but not extranuclear genotype, also influenced the titre of Rickettsia. Our results support the hypothesis that differences in host nuclear genotype alone may explain considerable within-population variation in host-symbiont phenotype and may contribute to the observed variation in Rickettsia-whitefly interactions worldwide.}, } @article {pmid27738153, year = {2016}, author = {Servick, K}, title = {Winged warriors.}, journal = {Science (New York, N.Y.)}, volume = {354}, number = {6309}, pages = {164-167}, doi = {10.1126/science.354.6309.164}, pmid = {27738153}, issn = {1095-9203}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Animals, Genetically Modified/*genetics ; Brazil ; Chikungunya Fever/prevention & control ; Communicable Disease Control/*methods ; Dengue/prevention & control ; Female ; Humans ; Male ; Mosquito Control/*methods ; Wolbachia ; Zika Virus Infection/prevention & control ; }, } @article {pmid27734894, year = {2016}, author = {Brown, AM and Wasala, SK and Howe, DK and Peetz, AB and Zasada, IA and Denver, DR}, title = {Genomic evidence for plant-parasitic nematodes as the earliest Wolbachia hosts.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {34955}, pmid = {27734894}, issn = {2045-2322}, mesh = {Animals ; Biological Evolution ; DNA, Bacterial/genetics ; Female ; Gene Library ; *Genome, Bacterial ; Genomics ; Homeostasis ; In Situ Hybridization, Fluorescence ; Iron/metabolism ; Likelihood Functions ; Male ; Nematoda/*microbiology ; Phenotype ; Phylogeny ; Plants/*parasitology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia, one of the most widespread endosymbionts, is a target for biological control of mosquito-borne diseases (malaria and dengue virus), and antibiotic elimination of infectious filarial nematodes. We sequenced and analyzed the genome of a new Wolbachia strain (wPpe) in the plant-parasitic nematode Pratylenchus penetrans. Phylogenomic analyses placed wPpe as the earliest diverging Wolbachia, suggesting two evolutionary invasions into nematodes. The next branches comprised strains in sap-feeding insects, suggesting Wolbachia may have first evolved as a nutritional mutualist. Genome size, protein content, %GC, and repetitive DNA allied wPpe with mutualistic Wolbachia, whereas gene repertoire analyses placed it between parasite (A, B) and mutualist (C, D, F) groups. Conservation of iron metabolism genes across Wolbachia suggests iron homeostasis as a potential factor in its success. This study enhances our understanding of this globally pandemic endosymbiont, highlighting genetic patterns associated with host changes. Combined with future work on this strain, these genomic data could help provide potential new targets for plant-parasitic nematode control.}, } @article {pmid27731327, year = {2017}, author = {Kemp, DJ and Thomson, FE and Edwards, W and Iturbe-Ormaetxe, I}, title = {Incomplete offspring sex bias in Australian populations of the butterfly Eurema hecabe.}, journal = {Heredity}, volume = {118}, number = {3}, pages = {284-292}, pmid = {27731327}, issn = {1365-2540}, mesh = {Animals ; Australia ; Butterflies/*genetics/microbiology ; Female ; Male ; Pedigree ; *Sex Ratio ; Wolbachia ; }, abstract = {Theory predicts unified sex ratios for most organisms, yet biases may be engendered by selfish genetic elements such as endosymbionts that kill or feminize individuals with male genotypes. Although rare, feminization is established for Wolbachia-infected Eurema butterflies. This paradigm is presently confined to islands in the southern Japanese archipelago, where feminized phenotypes produce viable all-daughter broods. Here, we characterize sex bias for E. hecabe in continental Australia. Starting with 186 wild-caught females, we reared >6000 F1-F3 progeny in pedigree designs that incorporated selective antibiotic treatments. F1 generations expressed a consistent bias across 2 years and populations that was driven by an ~5% incidence of broods comprising ⩾80% daughters. Females from biased lineages continued to overproduce daughters over two generations of outcrossing to wild males. Treatment with antibiotics of differential strength influenced sex ratio only in biased lineages by inducing an equivalent incomplete degree of son overproduction. Brood sex ratios were nevertheless highly variable within lineages and across generations. Intriguingly, the cytogenetic signature of female karyotype was uniformly absent, even among phenotypic females in unbiased lineages. Molecular evidence supported the existence of a single Wolbachia strain at high prevalence, yet this was not clearly linked to brood sex bias. In sum, we establish an inherited, experimentally reversible tendency for incomplete offspring bias. Key features of our findings clearly depart from the Japanese feminization paradigm and highlight the potential for more subtle degrees of sex distortion in arthropods.}, } @article {pmid27727237, year = {2016}, author = {Bordenstein, SR and Bordenstein, SR}, title = {Eukaryotic association module in phage WO genomes from Wolbachia.}, journal = {Nature communications}, volume = {7}, number = {}, pages = {13155}, pmid = {27727237}, issn = {2041-1723}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {Ankyrin Repeat ; Bacteriophages/*genetics ; Conserved Sequence ; Eukaryota/*metabolism ; Furin/metabolism ; Gene Transfer, Horizontal ; Genes, Viral ; *Genome, Viral ; Models, Biological ; Phylogeny ; Spider Venoms/chemistry ; Tetratricopeptide Repeat ; Viral Proteins/metabolism ; Wolbachia/*virology ; }, abstract = {Viruses are trifurcated into eukaryotic, archaeal and bacterial categories. This domain-specific ecology underscores why eukaryotic viruses typically co-opt eukaryotic genes and bacteriophages commonly harbour bacterial genes. However, the presence of bacteriophages in obligate intracellular bacteria of eukaryotes may promote DNA transfers between eukaryotes and bacteriophages. Here we report a metagenomic analysis of purified bacteriophage WO particles of Wolbachia and uncover a eukaryotic association module in the complete WO genome. It harbours predicted domains, such as the black widow latrotoxin C-terminal domain, that are uninterrupted in bacteriophage genomes, enriched with eukaryotic protease cleavage sites and combined with additional domains to forge one of the largest bacteriophage genes to date (14,256 bp). To the best of our knowledge, these eukaryotic-like domains have never before been reported in packaged bacteriophages and their phylogeny, distribution and sequence diversity imply lateral transfers between bacteriophage/prophage and animal genomes. Finally, the WO genome sequences and identification of attachment sites will potentially advance genetic manipulation of Wolbachia.}, } @article {pmid27726483, year = {2017}, author = {Leite, GL and de Paulo, PD and Zanuncio, JC and Tavares, WS and Alvarenga, AC and Dourado, LR and Bispo, EP and Soares, MA}, title = {Herbicide toxicity, selectivity and hormesis of nicosulfuron on 10 Trichogrammatidae (Hymenoptera) species parasitizing Anagasta (= Ephestia) kuehniella (Lepidoptera: Pyralidae) eggs.}, journal = {Journal of environmental science and health. Part. B, Pesticides, food contaminants, and agricultural wastes}, volume = {52}, number = {1}, pages = {70-76}, doi = {10.1080/03601234.2016.1229476}, pmid = {27726483}, issn = {1532-4109}, mesh = {Animals ; Female ; Herbicides/*toxicity ; Hormesis/*drug effects ; Hymenoptera/*drug effects ; Lepidoptera/drug effects/*parasitology ; Male ; Moths ; Ovum/drug effects/parasitology ; Pest Control, Biological ; Pyridines/*toxicity ; Sex Ratio ; Sulfonylurea Compounds/*toxicity ; }, abstract = {Selective agrochemicals including herbicides that do not affect non-target organisms such as natural enemies are important in the integrated pest management (IPM) programs. The aim of this study was to evaluate the herbicide toxicity, selectivity and hormesis of nicosulfuron, recommended for the corn Zea mays L. (Poaceae) crop, on 10 Trichogrammatidae (Hymenoptera) species. A female of each Trichogramma spp. or Trichogrammatoidea annulata De Santis, 1972 was individually placed in plastic test tubes (no choice) with a cardboard containing 45 flour moth Anagasta (= Ephestia) kuehniella Zeller, 1879 (Lepidoptera: Pyralidae) eggs. Parasitism by these natural enemies was allowed for 48 h and the cardboards were sprayed with the herbicide nicosulfuron at 1.50 L.ha[-1], along with the control (only distilled water). Nicosulfuron reduced the emergence rate of Trichogramma bruni Nagaraja, 1983 females, but increased that of Trichogramma pretiosum Riley, 1879, Trichogramma acacioi Brun, Moraes and Smith, 1984 and T. annulata females. Conversely, this herbicide increased the emergence rate of Trichogramma brasiliensis Ashmead, 1904, T. bruni, Trichogramma galloi Zucchi, 1988 and Trichogramma soaresi Nagaraja, 1983 males and decreased those of T. acacioi, Trichogramma atopovilia Oatman and Platner, 1983 and T. pretiosum males. In addition, nicosulfuron reduced the sex ratio of T. galloi, Trichogramma bennetti Nagaraja and Nagarkatti, 1973 and T. pretiosum and increased that of T. acacioi, T. bruni, T. annulata, Trichogramma demoraesi Nagaraja, 1983, T. soaresi and T. brasiliensis. The herbicide nicosulfuron was "harmless" (class 1, <30% reduction) for females and the sex ratio of all Trichogrammatidae species based on the International Organization for Biological Control (IOBC) classification. The possible hormesis effect of nicosulfuron on Trichogrammatidae species and on the bacterium Wolbachia sp. (Rickettsiales: Rickettsiaceae) was also discussed.}, } @article {pmid27721807, year = {2016}, author = {Dittmer, J and van Opstal, EJ and Shropshire, JD and Bordenstein, SR and Hurst, GD and Brucker, RM}, title = {Disentangling a Holobiont - Recent Advances and Perspectives in Nasonia Wasps.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1478}, pmid = {27721807}, issn = {1664-302X}, abstract = {The parasitoid wasp genus Nasonia (Hymenoptera: Chalcidoidea) is a well-established model organism for insect development, evolutionary genetics, speciation, and symbiosis. The host-microbiota assemblage which constitutes the Nasonia holobiont (a host together with all of its associated microbes) consists of viruses, two heritable bacterial symbionts and a bacterial community dominated in abundance by a few taxa in the gut. In the wild, all four Nasonia species are systematically infected with the obligate intracellular bacterium Wolbachia and can additionally be co-infected with Arsenophonus nasoniae. These two reproductive parasites have different transmission modes and host manipulations (cytoplasmic incompatibility vs. male-killing, respectively). Pioneering studies on Wolbachia in Nasonia demonstrated that closely related Nasonia species harbor multiple and mutually incompatible Wolbachia strains, resulting in strong symbiont-mediated reproductive barriers that evolved early in the speciation process. Moreover, research on host-symbiont interactions and speciation has recently broadened from its historical focus on heritable symbionts to the entire microbial community. In this context, each Nasonia species hosts a distinguishable community of gut bacteria that experiences a temporal succession during host development and members of this bacterial community cause strong hybrid lethality during larval development. In this review, we present the Nasonia species complex as a model system to experimentally investigate questions regarding: (i) the impact of different microbes, including (but not limited to) heritable endosymbionts, on the extended phenotype of the holobiont, (ii) the establishment and regulation of a species-specific microbiota, (iii) the role of the microbiota in speciation, and (iv) the resilience and adaptability of the microbiota in wild populations subjected to different environmental pressures. We discuss the potential for easy microbiota manipulations in Nasonia as a promising experimental approach to address these fundamental aspects.}, } @article {pmid27721806, year = {2016}, author = {Bouchon, D and Zimmer, M and Dittmer, J}, title = {The Terrestrial Isopod Microbiome: An All-in-One Toolbox for Animal-Microbe Interactions of Ecological Relevance.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1472}, pmid = {27721806}, issn = {1664-302X}, abstract = {Bacterial symbionts represent essential drivers of arthropod ecology and evolution, influencing host traits such as nutrition, reproduction, immunity, and speciation. However, the majority of work on arthropod microbiota has been conducted in insects and more studies in non-model species across different ecological niches will be needed to complete our understanding of host-microbiota interactions. In this review, we present terrestrial isopod crustaceans as an emerging model organism to investigate symbiotic associations with potential relevance to ecosystem functioning. Terrestrial isopods comprise a group of crustaceans that have evolved a terrestrial lifestyle and represent keystone species in terrestrial ecosystems, contributing to the decomposition of organic matter and regulating the microbial food web. Since their nutrition is based on plant detritus, it has long been suspected that bacterial symbionts located in the digestive tissues might play an important role in host nutrition via the provisioning of digestive enzymes, thereby enabling the utilization of recalcitrant food compounds (e.g., cellulose or lignins). If this were the case, then (i) the acquisition of these bacteria might have been an important evolutionary prerequisite for the colonization of land by isopods, and (ii) these bacterial symbionts would directly mediate the role of their hosts in ecosystem functioning. Several bacterial symbionts have indeed been discovered in the midgut caeca of terrestrial isopods and some of them might be specific to this group of animals (i.e., Candidatus Hepatoplasma crinochetorum, Candidatus Hepatincola porcellionum, and Rhabdochlamydia porcellionis), while others are well-known intracellular pathogens (Rickettsiella spp.) or reproductive parasites (Wolbachia sp.). Moreover, a recent investigation of the microbiota in Armadillidium vulgare has revealed that this species harbors a highly diverse bacterial community which varies between host populations, suggesting an important share of environmental microbes in the host-associated microbiota. In this review, we synthesize our current knowledge on the terrestrial isopod microbiome and identify future directions to (i) fully understand the functional roles of particular bacteria (both intracellular or intestinal symbionts and environmental gut passengers), and (ii) whether and how the host-associated microbiota could influence the performance of terrestrial isopods as keystone species in soil ecosystems.}, } @article {pmid27713560, year = {2016}, author = {Kumar, N and Lin, M and Zhao, X and Ott, S and Santana-Cruz, I and Daugherty, S and Rikihisa, Y and Sadzewicz, L and Tallon, LJ and Fraser, CM and Dunning Hotopp, JC}, title = {Efficient Enrichment of Bacterial mRNA from Host-Bacteria Total RNA Samples.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {34850}, pmid = {27713560}, issn = {2045-2322}, support = {DP2 OD007372/OD/NIH HHS/United States ; R01 AI047885/AI/NIAID NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/microbiology ; Drosophila/microbiology ; Ehrlichia chaffeensis/genetics ; *Genetic Techniques ; High-Throughput Nucleotide Sequencing/methods ; Host-Pathogen Interactions/*genetics ; Microfluidic Analytical Techniques ; Poly A/chemistry ; RNA, Bacterial/chemistry/*isolation & purification ; RNA, Messenger/chemistry/*isolation & purification ; RNA, Ribosomal/chemistry ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Symbiosis ; Wolbachia/genetics/pathogenicity ; Wolves/microbiology ; }, abstract = {Despite numerous advances in genomics and bioinformatics, technological hurdles remain to examine host-microbe transcriptomics. Sometimes the transcriptome of either or both can be ascertained merely by generating more sequencing reads. However, many cases exist where bacterial mRNA needs to be enriched further to enable cost-effective sequencing of the pathogen or endosymbiont. While a suitable method is commercially available for mammalian samples of this type, development of such methods has languished for invertebrate samples. Furthermore, a common method across multiple taxa would facilitate comparisons between bacteria in invertebrate vectors and their vertebrate hosts. Here, a method is described to concurrently remove polyadenylated transcripts, prokaryotic rRNA, and eukaryotic rRNA, including those with low amounts of starting material (e.g. 100 ng). In a Wolbachia-Drosophila system, this bacterial mRNA enrichment yielded a 3-fold increase in Wolbachia mRNA abundance and a concomitant 3.3-fold increase in the percentage of transcripts detected. More specifically, 70% of the genome could be recovered by transcriptome sequencing compared to 21% in the total RNA. Sequencing of similar bacterial mRNA-enriched samples generated from Ehrlichia-infected canine cells covers 93% of the Ehrlichia genome, suggesting ubiquitous transcription across the entire Ehrlichia chaffeensis genome. This technique can potentially be used to enrich bacterial mRNA in many studies of host-microbe interactions.}, } @article {pmid27708633, year = {2016}, author = {Chen, S and Zhao, J and Joshi, D and Xi, Z and Norman, B and Walker, ED}, title = {Persistent Infection by Wolbachia wAlbB Has No Effect on Composition of the Gut Microbiota in Adult Female Anopheles stephensi.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1485}, pmid = {27708633}, issn = {1664-302X}, support = {R37 AI021884/AI/NIAID NIH HHS/United States ; }, abstract = {The bacteria in the midgut of Anopheles stephensi adult females from laboratory colonies were studied by sequencing the V4 region of 16S rRNA genes, with respect to three experimental factors: stable or cured Wolbachia infection; sugar or blood diet; and age. Proteobacteria and Bacteroidetes dominated the community [>90% of operational taxonomic units (OTUs)]; most taxa were in the classes Flavobacteriia, Gammaproteobacteria, and Alphaproteobacteria, and were assigned to Elizabethkingia (46.9%), Asaia (6.4%) and Pseudomonas (6.0%), or unclassified Enterobacteriaceae (37.2%). Bacterial communities were similar between Wolbachia-cured and Wolbachia-infected mosquito lines, indicating that the gut microbiota were not dysregulated in the presence of Wolbachia. The proportion of Enterobacteriaceae was higher in mosquitoes fed a blood meal compared to those provided a sugar meal. Collectively, the bacterial community had a similar structure in older Wolbachia-infected mosquitoes 8 days after the blood meal, as in younger Wolbachia-infected mosquitoes before a blood meal, except that older mosquitoes had a higher proportion of Enterobacteriaceae and lower proportion of Elizabethkingia. Consistent presence of certain predominant bacteria (Elizabethkingia, Asaia, Pseudomonas, and Enterobacteriaceae) suggests they would be useful for paratransgenesis to control malaria infection, particularly when coupled to a Wolbachia-based intervention strategy.}, } @article {pmid27707863, year = {2016}, author = {van Nouhuys, S and Kohonen, M and Duplouy, A}, title = {Wolbachia increases the susceptibility of a parasitoid wasp to hyperparasitism.}, journal = {The Journal of experimental biology}, volume = {219}, number = {Pt 19}, pages = {2984-2990}, doi = {10.1242/jeb.140699}, pmid = {27707863}, issn = {1477-9145}, mesh = {Animals ; Butterflies/parasitology ; Disease Resistance ; Estonia ; Finland ; Host-Pathogen Interactions ; Larva/parasitology ; Parasites/*microbiology ; Species Specificity ; Virulence ; Wasps/*microbiology/pathogenicity ; Wolbachia/*physiology ; }, abstract = {The success of maternally transmitted endosymbiotic bacteria, such as Wolbachia, is directly linked to their host reproduction but in direct conflict with other parasites that kill the host before it reaches reproductive maturity. Therefore, symbionts that have evolved strategies to increase their host's ability to evade lethal parasites may have high penetrance, while detrimental symbionts would be selected against, leading to lower penetrance or extinction from the host population. In a natural population of the parasitoid wasp Hyposoter horticola in the Åland Islands (Finland), the Wolbachia strain wHho persists at an intermediate prevalence (∼50%). Additionally, there is a negative correlation between the prevalence of Wolbachia and a hyperparasitoid wasp, Mesochorus cf. stigmaticus, in the landscape. Using a manipulative field experiment, we addressed the persistence of Wolbachia at this intermediate level, and tested whether the observed negative correlation could be due to Wolbachia inducing either susceptibility or resistance to parasitism. We show that infection with Wolbachia does not influence the ability of the wasp to parasitize its butterfly host, Melitaea cinxia, but that hyperparasitism of the wasp increases in the presence of wHho. Consequently, the symbiont is detrimental, and in order to persist in the host population, must also have a positive effect on fitness that outweighs the costly burden of susceptibility to widespread parasitism.}, } @article {pmid27706105, year = {2016}, author = {Benelli, G and Jeffries, CL and Walker, T}, title = {Biological Control of Mosquito Vectors: Past, Present, and Future.}, journal = {Insects}, volume = {7}, number = {4}, pages = {}, pmid = {27706105}, issn = {2075-4450}, support = {//Wellcome Trust/United Kingdom ; 101285//Wellcome Trust/United Kingdom ; }, abstract = {Mosquitoes represent the major arthropod vectors of human disease worldwide transmitting malaria, lymphatic filariasis, and arboviruses such as dengue virus and Zika virus. Unfortunately, no treatment (in the form of vaccines or drugs) is available for most of these diseases andvectorcontrolisstillthemainformofprevention. Thelimitationsoftraditionalinsecticide-based strategies, particularly the development of insecticide resistance, have resulted in significant efforts to develop alternative eco-friendly methods. Biocontrol strategies aim to be sustainable and target a range of different mosquito species to reduce the current reliance on insecticide-based mosquito control. In thisreview, weoutline non-insecticide basedstrategiesthat havebeenimplemented orare currently being tested. We also highlight the use of mosquito behavioural knowledge that can be exploited for control strategies.}, } @article {pmid27703857, year = {2016}, author = {García-Vázquez, D and Ribera, I}, title = {The origin of widespread species in a poor dispersing lineage (diving beetle genus Deronectes).}, journal = {PeerJ}, volume = {4}, number = {}, pages = {e2514}, pmid = {27703857}, issn = {2167-8359}, abstract = {In most lineages, most species have restricted geographic ranges, with only few reaching widespread distributions. How these widespread species reached their current ranges is an intriguing biogeographic and evolutionary question, especially in groups known to be poor dispersers. We reconstructed the biogeographic and temporal origin of the widespread species in a lineage with particularly poor dispersal capabilities, the diving beetle genus Deronectes (Dytiscidae). Most of the ca. 60 described species of Deronectes have narrow ranges in the Mediterranean area, with only four species with widespread European distributions. We sequenced four mitochondrial and two nuclear genes of 297 specimens of 109 different populations covering the entire distribution of the four lineages of Deronectes, including widespread species. Using Bayesian probabilities with an a priori evolutionary rate, we performed (1) a global phylogeny/phylogeography to estimate the relationships of the main lineages within each group and root them, and (2) demographic analyses of the best population coalescent model for each species group, including a reconstruction of the geographical history estimated from the distribution of the sampled localities. We also selected 56 specimens to test for the presence of Wolbachia, a maternally transmitted parasite that can alter the patterns of mtDNA variability. All species of the four studied groups originated in the southern Mediterranean peninsulas and were estimated to be of Pleistocene origin. In three of the four widespread species, the central and northern European populations were nested within those in the northern areas of the Anatolian, Balkan and Iberian peninsulas respectively, suggesting a range expansion at the edge of the southern refugia. In the Mediterranean peninsulas the widespread European species were replaced by vicariant taxa of recent origin. The fourth species (D. moestus) was proven to be a composite of unrecognised lineages with more restricted distributions around the Western and Central Mediterranean. The analysis of Wolbachia showed a high prevalence of infection among Deronectes, especially in the D. aubei group, where all sequenced populations were infected with the only exception of the Cantabrian Mountains, the westernmost area of distribution of the lineage. In this group there was a phylogenetic incongruence between the mitochondrial and the nuclear sequence, although no clear pattern links this discordance to the Wolbachia infection. Our results suggest that, in different glacial cycles, populations that happened to be at the edge of the newly deglaciated areas took advantage of the optimal ecological conditions to expand their ranges to central and northern Europe. Once this favourable ecological window ended populations become isolated, resulting in the presence of closely related but distinct species in the Mediterranean peninsulas.}, } @article {pmid27693536, year = {2016}, author = {Kuesel, AC}, title = {Research for new drugs for elimination of onchocerciasis in Africa.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {6}, number = {3}, pages = {272-286}, pmid = {27693536}, issn = {2211-3207}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Africa/epidemiology ; Animals ; *Disease Eradication ; Drug Approval ; Drug Discovery/*trends ; Drug Therapy/*methods ; Filaricides/*isolation & purification/*therapeutic use ; Onchocerciasis/*epidemiology/*prevention & control ; }, abstract = {Onchocerciasis is a parasitic, vector borne disease caused by the filarial nematode Onchocerca volvulus. More than 99% of the population at risk of infection live in Africa. Onchocerciasis control was initiated in West Africa in 1974 with vector control, later complemented by ivermectin mass drug administration and in the other African endemic countries in 1995 with annual community directed treatment with ivermectin (CDTI.) This has significantly reduced infection prevalence. Together with proof-of-concept for onchocerciasis elimination with annual CDTI from foci in Senegal and Mali, this has resulted in targeting onchocerciasis elimination in selected African countries by 2020 and in 80% of African countries by 2025. The challenges for meeting these targets include the number of endemic countries where conflict has delayed or interrupted control programmes, cross-border foci, potential emergence of parasite strains with low susceptibility to ivermectin and co-endemicity of loiasis, another parasitic vector borne disease, which slows down or prohibits CDTI implementation. Some of these challenges could be addressed with new drugs or drug combinations with a higher effect on Onchocerca volvulus than ivermectin. This paper reviews the path from discovery of new compounds to their qualification for large scale use and the support regulatory authorities provide for development of drugs for neglected tropical diseases. The status of research for new drugs or treatment regimens for onchocerciasis along the path to regulatory approval and qualification for large scale use is reviewed. This research includes new regimens and combinations of ivermectin and albendazole, antibiotics targeting the O. volvulus endosymbiont Wolbachia, flubendazole, moxidectin and emodepside and discovery of new compounds.}, } @article {pmid27692043, year = {2016}, author = {Schnettler, E and Sreenu, VB and Mottram, T and McFarlane, M}, title = {Wolbachia restricts insect-specific flavivirus infection in Aedes aegypti cells.}, journal = {The Journal of general virology}, volume = {97}, number = {11}, pages = {3024-3029}, pmid = {27692043}, issn = {1465-2099}, support = {MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/1/MRC_/Medical Research Council/United Kingdom ; G0801822/MRC_/Medical Research Council/United Kingdom ; MC_UP_A550_1031/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/12/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Drosophila/microbiology ; Flavivirus/*physiology ; Insect Vectors/microbiology/virology ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Mosquito-borne viruses are known to cause disease in humans and livestock and are often difficult to control due to the lack of specific antivirals and vaccines. The Wolbachia endosymbiont has been widely studied for its ability to restrict positive-strand RNA virus infection in mosquitoes, although little is known about the precise antiviral mechanism. In recent years, a variety of insect-specific viruses have been discovered in mosquitoes and an interaction with mosquito-borne viruses has been reported for some of them; however, nothing is known about the effect of Wolbachia on insect-specific virus infection in mosquitoes. Here, we show that transinfection of the Drosophila-derived wMelPop Wolbachia strain into Aedes aegypti-derived cells resulted in inhibition and even clearance of the persistent cell-fusing agent flavivirus infection in these cells. This broadens the antiviral activity of Wolbachia from acute infections to persistent infections and from arboviruses to mosquito-specific viruses. In contrast, no effect on the Phasi Charoen-like bunyavirus persistent infection in these cells was observed, suggesting a difference in Wolbachia inhibition between positive- and negative-strand RNA viruses.}, } @article {pmid30387572, year = {2016}, author = {Rakova, VM}, title = {DIROFILARIASIS: CURRENT ASPECTS OF STUDIES.}, journal = {Meditsinskaia parazitologiia i parazitarnye bolezni}, volume = {4}, number = {4}, pages = {48-52}, pmid = {30387572}, issn = {0025-8326}, mesh = {Animals ; Dirofilaria/classification/*physiology ; Dirofilariasis/*epidemiology/*parasitology/therapy ; Europe/epidemiology ; *Geographic Information Systems ; Humans ; }, abstract = {The review touches upon the main directions and some of the most important results of an investigation of dirofilariasis the species of the genus Dirofilaria, their spread, and hosts. It describes cases of human infection with Candidatus Dirofilaria hongkongensis, a study of the prevalence of filariasis using a geographic information system, data on the contamination of Dirofilaria in Europe, and current views on the endosymbiotic relations of Dirofilaria with the Wolbachia bacterium.}, } @article {pmid27684942, year = {2016}, author = {Faria, VG and Martins, NE and Magalhães, S and Paulo, TF and Nolte, V and Schlötterer, C and Sucena, É and Teixeira, L}, title = {Drosophila Adaptation to Viral Infection through Defensive Symbiont Evolution.}, journal = {PLoS genetics}, volume = {12}, number = {9}, pages = {e1006297}, pmid = {27684942}, issn = {1553-7404}, support = {P 27630/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Microbial symbionts can modulate host interactions with biotic and abiotic factors. Such interactions may affect the evolutionary trajectories of both host and symbiont. Wolbachia protects Drosophila melanogaster against several viral infections and the strength of the protection varies between variants of this endosymbiont. Since Wolbachia is maternally transmitted, its fitness depends on the fitness of its host. Therefore, Wolbachia populations may be under selection when Drosophila is subjected to viral infection. Here we show that in D. melanogaster populations selected for increased survival upon infection with Drosophila C virus there is a strong selection coefficient for specific Wolbachia variants, leading to their fixation. Flies carrying these selected Wolbachia variants have higher survival and fertility upon viral infection when compared to flies with the other variants. These findings demonstrate how the interaction of a host with pathogens shapes the genetic composition of symbiont populations. Furthermore, host adaptation can result from the evolution of its symbionts, with host and symbiont functioning as a single evolutionary unit.}, } @article {pmid27681542, year = {2016}, author = {Mohanty, I and Rath, A and Mahapatra, N and Hazra, RK}, title = {Wolbachia: A biological control strategy against arboviral diseases.}, journal = {Journal of vector borne diseases}, volume = {53}, number = {3}, pages = {199-207}, pmid = {27681542}, issn = {0972-9062}, mesh = {Animals ; Antibiosis ; Arbovirus Infections/epidemiology/*prevention & control ; Arboviruses/*growth & development ; Culicidae/*microbiology ; Disease Transmission, Infectious/*prevention & control ; Humans ; Mosquito Control/*methods ; Wolbachia/*growth & development ; }, abstract = {Vector-borne diseases particularly those transmitted by mosquitoes like Dengue are among the leading causes of mortality and morbidity in human population. There are no effective vaccines or treatment against dengue fever till date and the control methods are limited. So, new approaches are urgently in need to reverse these trends. Vector control is currently the primary intervention tool. Strategies that reduce or block pathogen transmission by mosquitoes have been proposed as a means of augmenting current control measures to reduce the growing burden of vector-borne diseases. Wolbachia an endosymbiont of arthropod vectors is being explored as a novel ecofriendly control strategy. Studies in Drosophila have shown that Wolbachia can confer resistance to diverse RNA viruses and protect flies from virus-induced mortality. This review was focused on biology of the Wolbachia and its implication as a control measure for arboviral diseases mainly Dengue and Chikungunya.}, } @article {pmid27680689, year = {2017}, author = {Gunadi, A and Bansal, R and Finer, JJ and Michel, A}, title = {Establishment of in vitro soybean aphids, Aphis glycines (Hemiptera: Aphididae): a tool to facilitate studies of aphid symbionts, plant-insect interactions and insecticide efficacy.}, journal = {Pest management science}, volume = {73}, number = {6}, pages = {1229-1235}, doi = {10.1002/ps.4448}, pmid = {27680689}, issn = {1526-4998}, mesh = {Animals ; Aphids/drug effects/microbiology/*physiology ; Buchnera ; Enterobacteriaceae ; Insecticides/*pharmacology ; Neonicotinoids/pharmacology ; Nitro Compounds/pharmacology ; Oxazines/pharmacology ; Glycine max/*physiology ; Symbiosis ; Thiamethoxam ; Thiazoles/pharmacology ; Wolbachia ; ortho-Aminobenzoates/pharmacology ; }, abstract = {BACKGROUND: Studies on plant-insect interactions of the soybean aphid, Aphis glycines (Matsumura), can be influenced by environmental fluctuations, status of the host plant and variability in microbial populations. Maintenance of aphids on in vitro-grown plants minimizes environmental fluctuations, provides uniform host materials and permits the selective elimination of aphid-associated microbes for more standardized controls in aphid research.

RESULTS: Aphids were reared on sterile, in vitro-grown soybean seedlings germinated on plant tissue culture media amended with a mixture of antimicrobials. For initiation and maintenance of in vitro aphid colonies, single aphids were inoculated onto single in vitro seedlings. After three rounds of transfer of 'clean' aphids to fresh in vitro seedlings, contamination was no longer observed, and aphids performed equally well when compared with those reared on detached leaves. The addition of the insecticides thiamethoxam and chlorantraniliprole to the culture medium confirmed uptake and caused significant mortality to the in vitro aphids. The use of the antimicrobial mixture removed the associated bacteria Arsenophonus but retained Buchnera and Wolbachia within the in vitro aphids.

CONCLUSION: The in vitro aphid system is a novel and highly useful tool to understand insecticidal efficacy and expand our knowledge of tritrophic interactions among plants, insects and symbionts. © 2016 Society of Chemical Industry.}, } @article {pmid27677378, year = {2016}, author = {Kohl, A and Pondeville, E and Schnettler, E and Crisanti, A and Supparo, C and Christophides, GK and Kersey, PJ and Maslen, GL and Takken, W and Koenraadt, CJ and Oliva, CF and Busquets, N and Abad, FX and Failloux, AB and Levashina, EA and Wilson, AJ and Veronesi, E and Pichard, M and Arnaud Marsh, S and Simard, F and Vernick, KD}, title = {Advancing vector biology research: a community survey for future directions, research applications and infrastructure requirements.}, journal = {Pathogens and global health}, volume = {110}, number = {4-5}, pages = {164-172}, pmid = {27677378}, issn = {2047-7732}, support = {323173/ERC_/European Research Council/International ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Arbovirus Infections/*prevention & control/transmission ; Arboviruses/physiology ; Arthropod Vectors/*physiology ; Culicidae/*physiology ; Europe ; Host-Pathogen Interactions ; Humans ; Malaria/*prevention & control/transmission ; *Pest Control, Biological ; Plasmodium/physiology ; Research ; Surveys and Questionnaires ; Ticks/*physiology ; Wolbachia/physiology ; }, abstract = {Vector-borne pathogens impact public health, animal production, and animal welfare. Research on arthropod vectors such as mosquitoes, ticks, sandflies, and midges which transmit pathogens to humans and economically important animals is crucial for development of new control measures that target transmission by the vector. While insecticides are an important part of this arsenal, appearance of resistance mechanisms is increasingly common. Novel tools for genetic manipulation of vectors, use of Wolbachia endosymbiotic bacteria, and other biological control mechanisms to prevent pathogen transmission have led to promising new intervention strategies, adding to strong interest in vector biology and genetics as well as vector-pathogen interactions. Vector research is therefore at a crucial juncture, and strategic decisions on future research directions and research infrastructure investment should be informed by the research community. A survey initiated by the European Horizon 2020 INFRAVEC-2 consortium set out to canvass priorities in the vector biology research community and to determine key activities that are needed for researchers to efficiently study vectors, vector-pathogen interactions, as well as access the structures and services that allow such activities to be carried out. We summarize the most important findings of the survey which in particular reflect the priorities of researchers in European countries, and which will be of use to stakeholders that include researchers, government, and research organizations.}, } @article {pmid27666778, year = {2017}, author = {Dahmani, M and Davoust, B and Rousseau, F and Raoult, D and Fenollar, F and Mediannikov, O}, title = {Natural Anaplasmataceae infection in Rhipicephalus bursa ticks collected from sheep in the French Basque Country.}, journal = {Ticks and tick-borne diseases}, volume = {8}, number = {1}, pages = {18-24}, doi = {10.1016/j.ttbdis.2016.09.009}, pmid = {27666778}, issn = {1877-9603}, mesh = {Anaplasmataceae/*physiology ; Animals ; Female ; France/epidemiology ; Male ; Rhipicephalus/*microbiology ; Sheep ; Sheep Diseases/epidemiology/*parasitology ; Tick Infestations/epidemiology/*veterinary ; }, abstract = {Rhipicephalus bursa is one of 79 species of the genus Rhipicephalus in the family of Ixodidae. In this study, we investigated Anaplasmataceae bacteria associated with R. bursa collected after an epizootic outbreak of ovine anaplasmosis. 76 adult ticks, (60 male and 16 female ticks), were removed from sheep in two farms and all identified as R. bursa, all females were partially engorged. We found that 50% of the ticks were positive in the initial Anaplasmataceae qPCR screening. Bacterial species was identified by analyzing the sequences of amplicons of 23S rRNA, groEL and rpoB genes. 22.4% of ticks contained DNA of Anaplasma phagocytophilum and 7.9% the DNA of Anaplasma ovis. Based on 23S rRNA and groEL genes analysis, we found that 19.7% of ticks contained a potentially new species of Ehrlichia. We propose the status of Candidatus for this uncultured species and we provisionally name it Candidatus Ehrlichia urmitei. No Wolbachia were identified. These results show that R. bursa can be a carrier of Anaplasmataceae bacteria.}, } @article {pmid27665524, year = {2016}, author = {Makepeace, BL and Tanya, VN}, title = {25 Years of the Onchocerca ochengi Model.}, journal = {Trends in parasitology}, volume = {32}, number = {12}, pages = {966-978}, doi = {10.1016/j.pt.2016.08.013}, pmid = {27665524}, issn = {1471-5007}, mesh = {Animals ; Host-Parasite Interactions ; Humans ; Insect Vectors/parasitology ; Onchocerca/physiology ; Onchocerciasis/drug therapy/*parasitology/*prevention & control ; Simuliidae/parasitology ; }, abstract = {Although of limited veterinary significance, Onchocerca ochengi has become famous as a natural model or 'analogue' of human onchocerciasis (river blindness), which is caused by Onchocerca volvulus. On the basis of both morphological and molecular criteria, O. ochengi is the closest extant relative of O. volvulus and shares several key natural history traits with the human pathogen. These include exploitation of the same group of insect vectors (blackflies of the Simulium damnosum complex) and formation of collagenous nodules with a similar histological structure to human nodules. Here, we review the contribution of this natural system to drug and vaccine discovery efforts, as well as to our basic biological understanding of Onchocerca spp., over the past quarter-century.}, } @article {pmid27661080, year = {2016}, author = {Rohrscheib, CE and Frentiu, FD and Horn, E and Ritchie, FK and van Swinderen, B and Weible, MW and O'Neill, SL and Brownlie, JC}, title = {Intensity of Mutualism Breakdown Is Determined by Temperature Not Amplification of Wolbachia Genes.}, journal = {PLoS pathogens}, volume = {12}, number = {9}, pages = {e1005888}, pmid = {27661080}, issn = {1553-7374}, abstract = {Wolbachia are maternally transmitted intracellular bacterial symbionts that infect approximately 40% of all insect species. Though several strains of Wolbachia naturally infect Drosophila melanogaster and provide resistance against viral pathogens, or provision metabolites during periods of nutritional stress, one virulent strain, wMelPop, reduces fly lifespan by half, possibly as a consequence of over-replication. While the mechanisms that allow wMelPop to over-replicate are still of debate, a unique tandem repeat locus in the wMelPop genome that contains eight genes, referred to as the "Octomom" locus has been identified and is thought to play an important regulatory role. Estimates of Octomom locus copy number correlated increasing copy number to both Wolbachia bacterial density and increased pathology. Here we demonstrate that infected fly pathology is not dependent on an increased Octomom copy number, but does strongly correlate with increasing temperature. When measured across developmental time, we also show Octomom copy number to be highly variable across developmental time within a single generation. Using a second pathogenic strain of Wolbachia, we further demonstrate reduced insect lifespan can occur independently of a high Octomom locus copy number. Taken together, this data demonstrates that the mechanism/s of wMelPop virulence is more complex than has been previously described.}, } @article {pmid27659038, year = {2016}, author = {Mains, JW and Brelsfoard, CL and Rose, RI and Dobson, SL}, title = {Female Adult Aedes albopictus Suppression by Wolbachia-Infected Male Mosquitoes.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {33846}, pmid = {27659038}, issn = {2045-2322}, support = {R43 AI098179/AI/NIAID NIH HHS/United States ; R44 AI098179/AI/NIAID NIH HHS/United States ; }, abstract = {Dengue, chikungunya and zika viruses are pathogens with an increasing global impact. In the absence of an approved vaccine or therapy, their management relies on controlling the mosquito vectors. But traditional controls are inadequate, and the range of invasive species such as Aedes albopictus (Asian Tiger Mosquito) is expanding. Genetically modified mosquitoes are being tested, but their use has encountered regulatory barriers and public opposition in some countries. Wolbachia bacteria can cause a form of conditional sterility, which can provide an alternative to genetic modification or irradiation. It is unknown however, whether openly released, artificially infected male Ae. albopictus can competitively mate and sterilize females at a level adequate to suppress a field population. Also, the unintended establishment of Wolbachia at the introduction site could result from horizontal transmission or inadvertent female release. In 2014, an Experimental Use Permit from the United States Environmental Protection Agency approved a pilot field trial in Lexington, Kentucky, USA. Here, we present data showing localized reduction of both egg hatch and adult female numbers. The artificial Wolbachia type was not observed to establish in the field. The results are discussed in relation to the applied use of Wolbachia-infected males as a biopesticide to suppress field populations of Ae. albopictus.}, } @article {pmid27648768, year = {2016}, author = {Mascarenhas, RO and Prezotto, LF and Perondini, AL and Marino, CL and Selivon, D}, title = {Wolbachia in guilds of Anastrepha fruit flies (Tephritidae) and parasitoid wasps (Braconidae).}, journal = {Genetics and molecular biology}, volume = {39}, number = {4}, pages = {600-610}, pmid = {27648768}, issn = {1415-4757}, abstract = {The endosymbiont Wolbachia is efficiently transmitted from females to their progenies, but horizontal transmission between different taxa is also known to occur. Aiming to determine if horizontal transmission might have occurred between Anastrepha fruit flies and associated braconid wasps, infection by Wolbachia was screened by amplification of a fragment of the wsp gene. Eight species of the genus Anastrepha were analyzed, from which six species of associated parasitoid wasps were recovered. The endosymbiont was found in seven Anastrepha species and in five species of braconids. The WSP Typing methodology detected eight wsp alleles belonging to Wolbachia supergroup A. Three were already known and five were new ones, among which four were found to be putative recombinant haplotypes. Two samples of Anastrepha obliqua and one sample of Doryctobracon brasiliensis showed multiple infection. Single infection by Wolbachia was found in the majority of samples. The distribution of Wolbachia harboring distinct alleles differed significantly between fruit flies and wasps. However, in nine samples of fruit flies and associated wasps, Wolbachia harbored the same wsp allele. These congruences suggest that horizontal transfer of Wolbachia might have occurred in the communities of fruit flies and their braconid parasitoids.}, } @article {pmid27628851, year = {2017}, author = {Xue, L and Manore, CA and Thongsripong, P and Hyman, JM}, title = {Two-sex mosquito model for the persistence of Wolbachia.}, journal = {Journal of biological dynamics}, volume = {11}, number = {sup1}, pages = {216-237}, doi = {10.1080/17513758.2016.1229051}, pmid = {27628851}, issn = {1751-3766}, mesh = {Aedes/*microbiology ; Animals ; Basic Reproduction Number ; Female ; Male ; *Models, Biological ; Wolbachia/*physiology ; }, abstract = {We develop and analyse an ordinary differential equation model to investigate the transmission dynamics of releasing Wolbachia-infected mosquitoes to establish an endemic infection in a population of wild uninfected mosquitoes. Wolbachia is a genus of endosymbiotic bacteria that can infect mosquitoes and reduce their ability to transmit some viral mosquito-transmitted diseases, including dengue fever, chikungunya, and Zika. Although the bacterium is transmitted vertically from infected mothers to their offspring, it can be difficult to establish an endemic infection in a wild mosquito population. Our transmission model for the adult and aquatic-stage mosquitoes takes into account Wolbachia-induced fitness change and cytoplasmic incompatibility. We show that, for a wide range of realistic parameter values, the basic reproduction number, [Formula: see text], is less than one. Hence, the epidemic will die out if only a few Wolbachia-infected mosquitoes are introduced into the wild population. Even though the basic reproduction number is less than one, an endemic Wolbachia infection can be established if a sufficient number of infected mosquitoes are released. This threshold effect is created by a backward bifurcation with three coexisting equilibria: a stable zero-infection equilibrium, an intermediate-infection unstable endemic equilibrium, and a high-infection stable endemic equilibrium. We analyse the impact of reducing the wild mosquito population before introducing the infected mosquitoes and observed that the most effective approach to establish the infection in the wild is based on reducing mosquitoes in both the adult and aquatic stages.}, } @article {pmid27623259, year = {2016}, author = {Sullivan, W}, title = {Endosymbiosis: The Remarkable Healing Powers of Wolbachia.}, journal = {Current biology : CB}, volume = {26}, number = {17}, pages = {R797-9}, doi = {10.1016/j.cub.2016.07.045}, pmid = {27623259}, issn = {1879-0445}, mesh = {Animals ; Arthropods ; Germ Cells ; Reproduction ; *Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia, a maternally transmitted bacterium globally present in arthropods, favors its own transmission by producing dramatic changes in host reproduction. Insight into the underlying molecular and cellular mechanisms comes from the identification of the Wolbachia effector protein TomO, which maintains host germline stem cells in an undifferentiated state.}, } @article {pmid27618991, year = {2016}, author = {Vivero, RJ and Jaramillo, NG and Cadavid-Restrepo, G and Soto, SI and Herrera, CX}, title = {Structural differences in gut bacteria communities in developmental stages of natural populations of Lutzomyia evansi from Colombia's Caribbean coast.}, journal = {Parasites & vectors}, volume = {9}, number = {1}, pages = {496}, pmid = {27618991}, issn = {1756-3305}, mesh = {Animal Distribution ; Animals ; Bacteria/*classification/*isolation & purification ; Colombia ; Female ; Gastrointestinal Tract/*microbiology ; Life Cycle Stages ; Male ; Psychodidae/*microbiology ; }, abstract = {BACKGROUND: Lutzomyia evansi, a phlebotomine insect endemic to Colombia's Caribbean coast, is considered to be the main vector of visceral and cutaneous leishmaniasis in the region. Although insects of this species can harbor pathogenic and non-pathogenic microorganisms in their intestinal microbiota, there is little information available about the diversity of gut bacteria present in Lutzomyia evansi. In this study, conventional microbiological methods and molecular tools were used to assess the composition of bacterial communities associated with Lutzomyia evansi guts in immature and adult stages of natural populations from the department of Sucre (Caribbean coast of Colombia).

METHODS: Sand flies were collected from two locations (peri-urban and jungle biotype) in the Department of Sucre (Caribbean coast of Colombia). A total of 752 Lutzomyia evansi intestines were dissected. In this study, 125 bacterial strains were isolated from different culture media (LB Agar, MacConkey Agar). Different methods were used for bacterial identification, including ribosomal intergenic spacer analysis (RISA) and analysis of the 16S rRNA and gyrB gene sequences. The genetic profiles of the bacterial populations were generated and temporal temperature gradient gel electrophoresis (TTGE) was used to compare them with total gut DNA. We also used PCR and DNA sequence analysis to determine the presence of Wolbachia endosymbiont bacteria and Leishmania parasites.

RESULTS: The culture-dependent technique showed that the dominant intestinal bacteria isolated belong to Acinetobacter, Enterobacter, Pseudomonas, Ochrobactrum, Shinella and Paenibacillus in the larval stage; Lysobacter, Microbacterium, Streptomyces, Bacillus and Rummeliibacillus in the pupal stage; and Staphylococcus, Streptomyces, Brevibacterium, Acinetobacter, Enterobacter and Pantoea in the adult stage. Statistical analysis revealed significant differences between the fingerprint patterns of the PCR-TTGE bands in bacterial communities from immature and adult stages. Additionally, differences were found in bacterial community structure in fed females, unfed females, males and larvae. The intestinal bacteria detected by PCR-TTGE were Enterobacter cloacae and Bacillus thuringiensis, which were present in different life stages of Lu. evansi, and Burkholderia cenocepacia and Bacillus gibsonii, which were detected only in the larval stage. Wolbachia and Leishmania were not detected in gut samples of Lutzomyia evansi.

CONCLUSIONS: The analyses conducted using microbiological and molecular approaches indicated significant variations in the bacterial communities associated with the gut of Lu. evansi, depending on the developmental stage and food source. We propose that these elements affect microbial diversity in L. evansi guts and may in turn influence pathogen transmission to humans bitten by this insect.}, } @article {pmid27611575, year = {2016}, author = {Shi, P and He, Z and Li, S and An, X and Lv, N and Ghanim, M and Cuthbertson, AG and Ren, SX and Qiu, BL}, title = {Wolbachia Has Two Different Localization Patterns in Whitefly Bemisia tabaci AsiaII7 Species.}, journal = {PloS one}, volume = {11}, number = {9}, pages = {e0162558}, pmid = {27611575}, issn = {1932-6203}, mesh = {Animals ; Eggs/microbiology ; Female ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Wolbachia/*physiology ; }, abstract = {The whitefly Bemisia tabaci is a cosmopolitan insect species complex that harbors the obligate primary symbiont Portiera aleyrodidarum and several facultative secondary symbionts including Wolbachia, which have diverse influences on the host biology. Here, for the first time, we revealed two different localization patterns of Wolbachia present in the immature and adult stages of B. tabaci AsiaII7 cryptic species. In the confined pattern, Wolbachia was restricted to the bacteriocytes, while in the scattered pattern Wolbachia localized in the bacteriocytes, haemolymph and other organs simultaneously. Our results further indicated that, the proportion of B. tabaci AsiaII7 individuals with scattered Wolbachia were significantly lower than that of confined Wolbachia, and the distribution patterns of Wolbachia were not associated with the developmental stage or sex of whitefly host. This study will provide a new insight into the various transmission routes of Wolbachia in different whitefly species.}, } @article {pmid27596624, year = {2016}, author = {Hunter, P}, title = {Challenges and options for disease vector control: The outbreak of Zika virus in South America and increasing insecticide resistance among mosquitoes have rekindled efforts for controlling disease vectors.}, journal = {EMBO reports}, volume = {17}, number = {10}, pages = {1370-1373}, pmid = {27596624}, issn = {1469-3178}, mesh = {Animals ; Culicidae/microbiology/*virology ; Disease Outbreaks/*prevention & control ; Gene Drive Technology ; Humans ; *Insecticide Resistance ; *Mosquito Control ; Mosquito Vectors/microbiology/*virology ; South America/epidemiology ; Wolbachia/physiology ; Zika Virus/physiology ; Zika Virus Infection/*epidemiology/prevention & control/transmission/virology ; }, abstract = {Vector control that targets disease‐transmitting insects has been used very efficiently to halt the spread of malaria and dengue fever. Increasing resistance to insecticides and new outbreaks of mosquito‐transmitted diseases has put the focus on both proven and novel sophisticated technologies to control disease vectors.[Image: see text]}, } @article {pmid27595920, year = {2016}, author = {Bajer, A and Rodo, A and Mierzejewska, EJ and Tołkacz, K and Welc-Faleciak, R}, title = {The prevalence of Dirofilaria repens in cats, healthy dogs and dogs with concurrent babesiosis in an expansion zone in central Europe.}, journal = {BMC veterinary research}, volume = {12}, number = {1}, pages = {183}, pmid = {27595920}, issn = {1746-6148}, mesh = {Animals ; Babesiosis/*complications/epidemiology ; Cat Diseases/epidemiology/*parasitology ; Cats ; Coinfection ; Dirofilaria repens/*isolation & purification ; Dirofilariasis/epidemiology/*parasitology ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Poland/epidemiology ; Prevalence ; Wolbachia/isolation & purification ; }, abstract = {BACKGROUND: Dirofilaria repens is a mosquito-transmitted, filarial nematode parasitizing dogs, cats and other carnivores. Recently, this parasite has spread in central Europe, including Poland. The aim of the present study was to estimate the prevalence of D. repens in cats and dogs in different regions of the country and to investigate the occurrence and consequences of co-infection with another fast-spreading vector-borne parasite, Babesia canis.

RESULTS: In the period 2013-2015, 147 blood samples from cats from central Poland and 257 blood samples from dogs from central, northern, southern and western Poland were collected. Prevalence of D. repens was determined by amplification and sequencing of the 12S rDNA gene fragment. Among dogs, 94 samples originated from clinically healthy dogs from central Poland (Masovia) and 58 samples originated from dogs that were infected with B. canis. Prevalence of D. repens was compared between these two groups of dogs. For the first time D. repens was identified in a cat from central Europe (0.7 % [95 % CL: 0-4.1 %]). The DNA of the filarial endosymbiotic bacterium Wolbachia was detected in two cats (1.4 % [95 % CL: 0-5.5 %]). In dogs, the parasite was detected only in samples from central Poland (Masovia) (local prevalence = 38 % [95 % CL: 25.9-51.8 %]). Prevalence of D. repens was significantly higher in dogs with babesiosis (90 % [95 % CL: 81.6-94.5 %]). Co-infections of D. repens and B. canis were confirmed by sequencing in 30 dogs with babesiosis, but no co-infections were identified in healthy dogs from Masovia. Statistical analyses of blood parameters revealed that dogs with co-infections suffered more severe anemia and thrombocytopenia, but presented milder changes in biochemical parameters (i.e. less elevated concentration of alkaline phosphatase [ALP] and serum urea) suggesting lower risk of hepatic or renal failure in comparison to dogs infected only with B. canis.

CONCLUSIONS: These findings are important due to the spread of dirofilariosis and babesiosis in central Europe, as microfilaraemic dogs seem to be more prone to babesiosis. The possible protective effect of the nematode infection against hepatic or renal failure in canine babesiosis and its mechanisms require further investigations.}, } @article {pmid27591564, year = {2016}, author = {Dumas, E and Atyame, CM and Malcolm, CA and Le Goff, G and Unal, S and Makoundou, P and Pasteur, N and Weill, M and Duron, O}, title = {Molecular data reveal a cryptic species within the Culex pipiens mosquito complex.}, journal = {Insect molecular biology}, volume = {25}, number = {6}, pages = {800-809}, doi = {10.1111/imb.12264}, pmid = {27591564}, issn = {1365-2583}, mesh = {Animals ; *Biological Evolution ; Cell Nucleus/genetics ; Culex/*classification/*genetics/growth & development/microbiology ; DNA/genetics ; DNA, Mitochondrial/genetics ; Haplotypes ; Larva/classification/genetics ; Multilocus Sequence Typing ; Phylogeny ; Pupa/classification/genetics ; Symbiosis ; Wolbachia/physiology ; }, abstract = {The Culex pipiens mosquito complex is a group of evolutionarily closely related species including C. pipiens and Culex quinquefasciatus, both infected by the cytoplasmically inherited Wolbachia symbiont. A Wolbachia-uninfected population of C. pipiens was however described in South Africa and was recently proposed to represent a cryptic species. In this study, we reconsidered the existence of this species by undertaking an extensive screening for the presence of Wolbachia-uninfected C. pipiens specimens and by characterizing their genetic relatedness with known members of the complex. We first report on the presence of Wolbachia-uninfected specimens in several breeding sites. We next confirm that these uninfected specimens unambiguously belong to the C. pipiens complex. Remarkably, all uninfected specimens harbour mitochondrial haplotypes that are either novel or identical to those previously found in South Africa. In all cases, these mitochondrial haplotypes are closely related, but different, to those found in other C. pipiens complex members known to be infected by Wolbachia. Altogether, these results corroborate the presence of a widespread cryptic species within the C. pipiens species complex. The potential role of this cryptic C. pipiens species in the transmission of pathogens remains however to be determined. The designation 'Culex juppi nov. sp.' is proposed for this mosquito species.}, } @article {pmid27570308, year = {2016}, author = {Saurav, GK and Daimei, G and Rana, VS and Popli, S and Rajagopal, R}, title = {Detection and Localization of Wolbachia in Thrips palmi Karny (Thysanoptera: Thripidae).}, journal = {Indian journal of microbiology}, volume = {56}, number = {2}, pages = {167-171}, pmid = {27570308}, issn = {0046-8991}, abstract = {Thrips palmi Karny is a globally distributed polyphagous agricultural pest. It causes huge economic loss by its biological behaviors like feeding, reproduction and transmission of tospoviruses. Since T. palmi shows close morphological similarities with other thrips species, we employed mitochondrial cytochrome oxidase 1 (mtCO1) gene as a molecular marker. BLAST analysis of this sequence helped us to identify the collected specimen as T. palmi. We observed the female to male ratio of about 3:1 from collected samples and suspected the presence of Wolbachia. The presence of Wolbachia was detected by PCR using genus specific primers of 16S rRNA gene. Further confirmation of Wolbachia strain was achieved by conducting PCR amplification of three ubiquitous genes ftsZ, gatB and groEL. A phylogenetic tree was constructed with concatenated sequences of ftsZ and gatB gene to assign supergroup to Wolbachia. Finally, we localized Wolbachia in abdominal region of the insect using fluorescent in situ hybridization with the help of confocal microscope. Our result confirmed the presence of Wolbachia supergroup B strain for the first time in T. palmi.}, } @article {pmid27567683, year = {2016}, author = {Ren, T and Wu, M}, title = {PhyloCore: A phylogenetic approach to identifying core taxa in microbial communities.}, journal = {Gene}, volume = {593}, number = {2}, pages = {330-333}, pmid = {27567683}, issn = {1879-0038}, support = {R01 GM108501/GM/NIGMS NIH HHS/United States ; }, mesh = {*Algorithms ; Microbiota/*genetics ; *Phylogeny ; }, abstract = {BACKGROUND: Identifying core microbiota is an important step for understanding the key components of microbial communities. Traditional approach that identifies core taxa at the OTU level ignores potential ecological coherence of higher rank taxa. There is a need to develop software that can systematically identify core taxa at and above the species level.

RESULTS: Here we developed PhyloCore, an application that uses a phylogeny-based algorithm to identify core taxa at the proper taxonomic levels. It incorporates a number of features that users can set according to their needs. Using multiple gut microbiota as test cases, we demonstrate that PhyloCore is more powerful and flexible than OTU-based approaches.

CONCLUSIONS: PhyloCore is a flexible and fast application that identifies core taxa at proper taxonomic levels, making it useful to sequence-based microbial ecology studies. The software is freely available at http://wolbachia.biology.virginia.edu/WuLab/Software.html.}, } @article {pmid27551975, year = {2017}, author = {Chen, F and Coates, B and He, KL and Bai, SX and Zhang, TT and Wang, ZY}, title = {Effects of Wolbachia on mitochondrial DNA variation in populations of Athetis lepigone (Lepidoptera: Noctuidae) in China.}, journal = {Mitochondrial DNA. Part A, DNA mapping, sequencing, and analysis}, volume = {28}, number = {6}, pages = {826-834}, doi = {10.1080/24701394.2016.1197216}, pmid = {27551975}, issn = {2470-1408}, mesh = {Animals ; Bacterial Infections/*genetics ; China ; *Genetic Variation ; Genome, Insect ; *Genome, Mitochondrial ; Haplotypes ; Moths/*genetics/microbiology ; Phylogeny ; Sequence Analysis, DNA ; *Wolbachia ; }, abstract = {Wolbachia are endosymbiotic bacteria that infect arthropods and incompatibility among strains can affect gene flow within host insect populations that can significantly result in host mitochondrial DNA (MtDNA) variation within or among populations. The effects of Wolbachia infection on Athetis lepigone MtDNA variation were studied across 12 geographical populations sampled from North China. Mitochondrial COI gene and Wolbachia wsp gene were both obtained from 423 individuals, including 46 Wolbachia infected and 377 uninfected individuals. Average A. lepigone infection rate was 11% (range: 0 to 26%), and wsp sequence variation defined three distinct Wolbachia strains; wLep1, wLep2, and wLep3, respectively. A haplotype network showed no relationship between A. lepigone MtDNA haplotype and Wolbachia infection. Furthermore, haplotype variation mainly exists within populations, although the proportion of variation is higher within uninfected compared to infected individuals. The lack of correlation between A. lepigone haplotype and Wolbachia strain, suggests symbiont infection may not affect mating compatibility or gene flow within host populations in China.}, } @article {pmid27543297, year = {2016}, author = {Ramírez-Puebla, ST and Ormeño-Orrillo, E and Vera-Ponce de León, A and Lozano, L and Sanchez-Flores, A and Rosenblueth, M and Martínez-Romero, E}, title = {Genomes of Candidatus Wolbachia bourtzisii wDacA and Candidatus Wolbachia pipientis wDacB from the Cochineal Insect Dactylopius coccus (Hemiptera: Dactylopiidae).}, journal = {G3 (Bethesda, Md.)}, volume = {6}, number = {10}, pages = {3343-3349}, pmid = {27543297}, issn = {2160-1836}, mesh = {Animals ; Bacterial Secretion Systems/genetics/metabolism ; Biological Transport ; Energy Metabolism ; Female ; Genetic Variation ; *Genome, Bacterial ; *Genomics/methods ; Hemiptera/*microbiology ; High-Throughput Nucleotide Sequencing ; RNA, Bacterial ; RNA, Ribosomal, 16S ; Stress, Physiological/genetics ; Symbiosis ; Virulence Factors ; Wolbachia/classification/*genetics/isolation & purification/metabolism ; }, abstract = {Dactylopius species, known as cochineal insects, are the source of the carminic acid dye used worldwide. The presence of two Wolbachia strains in Dactylopius coccus from Mexico was revealed by PCR amplification of wsp and sequencing of 16S rRNA genes. A metagenome analysis recovered the genome sequences of Candidatus Wolbachia bourtzisii wDacA (supergroup A) and Candidatus Wolbachia pipientis wDacB (supergroup B). Genome read coverage, as well as 16S rRNA clone sequencing, revealed that wDacB was more abundant than wDacA. The strains shared similar predicted metabolic capabilities that are common to Wolbachia, including riboflavin, ubiquinone, and heme biosynthesis, but lacked other vitamin and cofactor biosynthesis as well as glycolysis, the oxidative pentose phosphate pathway, and sugar uptake systems. A complete tricarboxylic acid cycle and gluconeogenesis were predicted as well as limited amino acid biosynthesis. Uptake and catabolism of proline were evidenced in Dactylopius Wolbachia strains. Both strains possessed WO-like phage regions and type I and type IV secretion systems. Several efflux systems found suggested the existence of metal toxicity within their host. Besides already described putative virulence factors like ankyrin domain proteins, VlrC homologs, and patatin-like proteins, putative novel virulence factors related to those found in intracellular pathogens like Legionella and Mycobacterium are highlighted for the first time in Wolbachia Candidate genes identified in other Wolbachia that are likely involved in cytoplasmic incompatibility were found in wDacB but not in wDacA.}, } @article {pmid27538532, year = {2016}, author = {Ferdy, JB and Liu, N and Sicard, M}, title = {Transmission modes and the evolution of feminizing symbionts.}, journal = {Journal of evolutionary biology}, volume = {29}, number = {12}, pages = {2395-2409}, doi = {10.1111/jeb.12963}, pmid = {27538532}, issn = {1420-9101}, mesh = {Animals ; *Biological Evolution ; Female ; Male ; *Reproduction ; Sex Characteristics ; Virulence ; *Wolbachia ; }, abstract = {Vertically transmitted symbionts can distort their host's reproduction to increase their own transmission. In Wolbachia and some other symbionts, a particular distortion of this sort is feminization, whereby genetic males, which cannot transmit symbionts, are converted during development into functional females, which do transmit symbionts when they reproduce. In this work, we propose a model to study how feminization intensity (i.e. penetrance) can evolve under different ecological constraints in WZ/ZZ hosts. More specifically, our model incorporates both imperfect vertical and horizontal transmission modes. The model shows that for most parameter values feminizing symbionts drive genetic females to extinction, which in turn favours the evolution of maximum feminization penetrance. Once genetic females are extinct, the actual value of feminization penetrance never depends on the efficiency of vertical transmission. Instead, the model shows that in conditions where the reproductive rate is high at demographic equilibrium, higher feminization levels are favoured. One consequence of this can be, for example, that evolutionarily stable feminization penetrance increases with the host's natural death rate, just as the virulence is predicted to do with the host's natural death rate in classic epidemiological models. Finally, we found that horizontal transmission had no impact on how feminization penetrance evolved when genetic females were extinct. However, horizontal transmission can permit genetic females to coexist with symbionts and, in this case, we demonstrate that the presence of genetic females selects symbionts for lower feminization penetrance.}, } @article {pmid27515668, year = {2016}, author = {Zurita, A and Gutiérrez, SG and Cutillas, C}, title = {Infection Rates of Wolbachia sp. and Bartonella sp. in Different Populations of Fleas.}, journal = {Current microbiology}, volume = {73}, number = {5}, pages = {704-713}, pmid = {27515668}, issn = {1432-0991}, mesh = {Animals ; Bartonella/genetics/isolation & purification/*physiology ; Cat Diseases/parasitology ; Cats ; Dog Diseases/parasitology ; Dogs ; Female ; Flea Infestations/parasitology/*veterinary ; Male ; Siphonaptera/*microbiology ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {In the present study, a molecular detection of Bartonella sp. and Wolbachia sp. in Ctenocephalides felis (Siphonaptera: Pulicidae) isolated from Canis lupus familiaris from different geographical areas of Spain, Iran and South Africa, and in Stenoponia tripectinata tripectinata isolated from Mus musculus from the Canary Islands has been carried out by amplification of the 16S ribosomal RNA partial gene of Wolbachia sp. and intergenic spacer region (its region) of Bartonella sp. A total of 70 % of C. felis analysed were infected by W. pipientis. This percentage of prevalence was considerably higher in female fleas than in male fleas. Bartonella DNA was not detected in C. felis from dogs, while Bartonella elizabethae was detected and identified in S. t. tripectinata from M. musculus from the Canary Islands representing 43.75 % prevalence. This report is the first to identify B. elizabethae in S. t. tripectinata collected in M. musculus from the Canary Islands. Thus, our results demonstrate that this flea is a potential vector of B. elizabethae and might play roles in human infection. The zoonotic character of this bartonellosis emphasizes the need to alert public health authorities and the veterinary community of the risk of infection.}, } @article {pmid27503142, year = {2016}, author = {Sim, AD and Wheeler, D}, title = {The venom gland transcriptome of the parasitoid wasp Nasonia vitripennis highlights the importance of novel genes in venom function.}, journal = {BMC genomics}, volume = {17}, number = {}, pages = {571}, pmid = {27503142}, issn = {1471-2164}, mesh = {Animals ; Cluster Analysis ; Computational Biology/methods ; Female ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Ovary/metabolism ; Sequence Analysis, RNA ; *Transcriptome ; Wasp Venoms/*genetics ; Wasps/*genetics ; }, abstract = {BACKGROUND: Prior to egg laying the parasitoid wasp Nasonia vitripennis envenomates its pupal host with a complex mixture of venom peptides. This venom induces several dramatic changes in the host, including developmental arrest, immunosuppression, and altered metabolism. The diverse and potent bioactivity of N. vitripennis venom provides opportunities for the development of novel acting pharmaceuticals based on these molecules. However, currently very little is known about the specific functions of individual venom peptides or what mechanisms underlie the hosts response to envenomation. Many of the venom peptides also lack bioinformatically derived annotations because no homologs can be identified in the sequences databases. The RNA interference system of N. vitripennis provides a method for functional characterisation of venom protein encoding genes, however working with the current list of 79 candidates represents a daunting task. For this reason we were interested in determining the expression levels of venom encoding genes in the venom gland, as this information could be used to rank candidates for further study. To do this we carried out deep transcriptome sequencing of the venom gland and ovary tissue and used RNA-seq to rank the venom protein encoding genes by expression level. The generation of a specific venom gland transcriptome dataset also provides further opportunities to investigate novel features of this specialised organ.

RESULTS: RNA-seq revealed that the highest expressed venom encoding gene in the venom gland was 'Venom protein Y'. The highest expressed annotated gene in this tissue was serine protease Nasvi2EG007167, which has previously been implicated in the apoptotic activity of N. vitripennis venom. As expected the RNA-seq confirmed that venom encoding genes are almost exclusively expressed in the venom gland relative to the neighbouring ovary tissue. Novel genes appear to perform key roles in N. vitripennis venom function, with over half of the 15 highest expressed venom encoding loci lacking bioinformatic annotations. The high throughput sequencing data also provided evidence for the existence of an additional 472 previously undescribed transcribed regions in the N. vitripennis genome. Finally, metatranscriptomic analysis of the venom gland transcriptome finds little evidence for the role of Wolbachia in the venom system.

CONCLUSIONS: The expression level information provided here for the N. vitripennis venom protein encoding genes represents a valuable dataset that can be used by the research community to rank candidates for further functional characterisation. These candidates represent bioactive peptides valuable in the development of new pharmaceuticals.}, } @article {pmid27498563, year = {2016}, author = {Ote, M and Ueyama, M and Yamamoto, D}, title = {Wolbachia Protein TomO Targets nanos mRNA and Restores Germ Stem Cells in Drosophila Sex-lethal Mutants.}, journal = {Current biology : CB}, volume = {26}, number = {17}, pages = {2223-2232}, doi = {10.1016/j.cub.2016.06.054}, pmid = {27498563}, issn = {1879-0445}, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Drosophila Proteins/*genetics/metabolism ; Drosophila melanogaster/*genetics/growth & development/microbiology ; Female ; *Gene Expression Regulation ; Germ Cells/metabolism ; RNA, Messenger/genetics/metabolism ; RNA-Binding Proteins/*genetics/metabolism ; Stem Cells/*metabolism ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia, endosymbiotic bacteria prevalent in invertebrates, manipulate their hosts in a variety of ways: they induce cytoplasmic incompatibility, male lethality, male-to-female transformation, and parthenogenesis. However, little is known about the molecular basis for host manipulation by these bacteria. In Drosophila melanogaster, Wolbachia infection makes otherwise sterile Sex-lethal (Sxl) mutant females capable of producing mature eggs. Through a functional genomic screen for Wolbachia genes with growth-inhibitory effects when expressed in cultured Drosophila cells, we identified the gene WD1278 encoding a novel protein we call toxic manipulator of oogenesis (TomO), which phenocopies some of the Wolbachia effects in Sxl mutant D. melanogaster females. We demonstrate that TomO enhances the maintenance of germ stem cells (GSCs) by elevating Nanos (Nos) expression via its interaction with nos mRNA, ultimately leading to the restoration of germ cell production in Sxl mutant females that are otherwise without GSCs.}, } @article {pmid27492563, year = {2016}, author = {Akhoundi, M and Cannet, A and Loubatier, C and Berenger, JM and Izri, A and Marty, P and Delaunay, P}, title = {Molecular characterization of Wolbachia infection in bed bugs (Cimex lectularius) collected from several localities in France.}, journal = {Parasite (Paris, France)}, volume = {23}, number = {}, pages = {31}, pmid = {27492563}, issn = {1776-1042}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Bedbugs/*microbiology ; DNA, Bacterial/chemistry/isolation & purification ; France ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia symbionts are maternally inherited intracellular bacteria that have been detected in numerous insects including bed bugs. The objective of this study, the first epidemiological study in Europe, was to screen Wolbachia infection among Cimex lectularius collected in the field, using PCR targeting the surface protein gene (wsp), and to compare obtained Wolbachia strains with those reported from laboratory colonies of C. lectularius as well as other Wolbachia groups. For this purpose, 284 bed bug specimens were caught and studied from eight different regions of France including the suburbs of Paris, Bouches-du-Rhône, Lot-et-Garonne, and five localities in Alpes-Maritimes. Among the samples, 166 were adults and the remaining 118 were considered nymphs. In all, 47 out of 118 nymphs (40%) and 61 out of 166 adults (37%) were found positive on wsp screening. Among the positive cases, 10 samples were selected randomly for sequencing. The sequences had 100% homology with wsp sequences belonging to the F-supergroup strains of Wolbachia. Therefore, we confirm the similarity of Wolbachia strains detected in this epidemiological study to Wolbachia spp. reported from laboratory colonies of C. lectularius.}, } @article {pmid27479050, year = {2016}, author = {Garcia, Gde A and Dos Santos, LM and Villela, DA and Maciel-de-Freitas, R}, title = {Using Wolbachia Releases to Estimate Aedes aegypti (Diptera: Culicidae) Population Size and Survival.}, journal = {PloS one}, volume = {11}, number = {8}, pages = {e0160196}, pmid = {27479050}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Animals ; Brazil ; Female ; *Models, Theoretical ; Pest Control, Biological ; Population Density ; Wolbachia/*physiology ; }, abstract = {Mosquitoes carrying the endosymbiont bacterium Wolbachia have been deployed in field trials as a biological control intervention due to Wolbachia effects on reducing transmission of arboviruses. We performed mark, release and recapture (MRR) experiments using Wolbachia as an internal marker with daily collections with BG-Traps during the first two weeks of releases in Rio de Janeiro, Brazil. The MRR design allowed us to investigate two critical parameters that determine whether Wolbachia would successful invade a field population: the probability of daily survival (PDS) of Wolbachia-infected Aedes aegypti females, and the wild population density during releases. Released females had a PDS of 0.82 and 0.89 in the first and second weeks, respectively, immediately after releases, which is well within the range of previous estimates of survivorship of wild mosquitoes in Rio de Janeiro. Abundance estimation of wild population varied up to 10-fold higher depending on the estimation method used (634-3565 females on the average-difference model to 6365-16188 females according to Lincoln-Petersen). Wolbachia-released mosquitoes were lower than the density estimation of their wild counterparts, irrespectively of the model used. Individually screening mosquitoes for the presence of Wolbachia reduced uncertainty on abundance estimations due to fluctuation in capturing per week. A successful invasion into local population requires Ae. aegypti fitness is unaffected by Wolbachia presence, but also reliable estimates on the population size of wild mosquitoes.}, } @article {pmid32260037, year = {2016}, author = {}, title = {Short news.}, journal = {Virologie (Montrouge, France)}, volume = {20}, number = {4}, pages = {248-252}, doi = {10.1684/vir.2016.0662}, pmid = {32260037}, issn = {1267-8694}, } @article {pmid27468908, year = {2016}, author = {Gofton, AW and Doggett, S and Ratchford, A and Ryan, U and Irwin, P}, title = {Phylogenetic characterisation of two novel Anaplasmataceae from Australian Ixodes holocyclus ticks: 'Candidatus Neoehrlichia australis' and 'Candidatus Neoehrlichia arcana'.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {66}, number = {10}, pages = {4256-4261}, doi = {10.1099/ijsem.0.001344}, pmid = {27468908}, issn = {1466-5034}, mesh = {Anaplasmataceae/*classification/genetics/isolation & purification ; Animals ; Australia ; Bacterial Typing Techniques ; Base Composition ; DNA, Bacterial/genetics ; Genes, Bacterial ; Ixodes/*microbiology ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {Recently, two novel species of Anaplasmataceae were detected in the Australian paralysis tick, Ixodes holocyclus, by 16S rRNA gene metabarcoding. Analysis of these sequences suggested that these novel organisms are closely related to the genus 'Candidatus Neoehrlichia'. In this study, phylogenetic analysis of 16S rRNA (1264 bp), groESL (1047 bp) and gltA (561 bp) gene sequences, and concatenated (2872 bp) sequences, all concur that these novel species belong in the genus 'Candidatus Neoehrlichia' and are most closely related to, but distinct from the only other recognised members of this genus, 'Candidatus Neoehrlichia mikurensis' and 'Candidatus Neoehrlichia lotoris'. Based on their unique molecular signature, we propose to designate these species 'Candidatus Neoehrlichia australis' (reference strain HT41R) and 'Candidatus Neoehrlichia arcana' (reference strain HT94R). Identical 'Candidatus Neoehrlichia australis' 16S rRNA, groESL and gltA sequences were detected in 34/391 (8.7 %) individual Ixodes holocyclus ticks, and sequences were most similar to 'Candidatus Neoehrlichia lotoris' (96.2 %, 83.1 % and 67.2 %, respectively) and 'Candidatus Neoehrlichia mikurensis' (96.2 %, 84 % and 68.4 % respectively). Likewise, identical 'Candidatus Neoehrlichia arcana' 16S rRNA, groESL and gltA sequences were detected in 12/391 (3.1 %) Ixodes holocyclus ticks, and sequences were most similar to 'Candidatus Neoehrlichia lotoris' (98.5 %, 88.7 % and 79.3 %, respectively) and 'Candidatus Neoehrlichia mikurensis' (96.3 %, 84 % and 67.4 % respectively). These new species are the first Anaplasmataceae (except Wolbachia spp.) to be found to be endemic to Australia. The pathogenic consequences of these organisms are yet to be determined.}, } @article {pmid27462300, year = {2016}, author = {Erban, T and Klimov, PB and Smrz, J and Phillips, TW and Nesvorna, M and Kopecky, J and Hubert, J}, title = {Populations of Stored Product Mite Tyrophagus putrescentiae Differ in Their Bacterial Communities.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1046}, pmid = {27462300}, issn = {1664-302X}, abstract = {BACKGROUND: Tyrophagus putrescentiae colonizes different human-related habitats and feeds on various post-harvest foods. The microbiota acquired by these mites can influence the nutritional plasticity in different populations. We compared the bacterial communities of five populations of T. putrescentiae and one mixed population of T. putrescentiae and T. fanetzhangorum collected from different habitats.

MATERIAL: The bacterial communities of the six mite populations from different habitats and diets were compared by Sanger sequencing of cloned 16S rRNA obtained from amplification with universal eubacterial primers and using bacterial taxon-specific primers on the samples of adults/juveniles or eggs. Microscopic techniques were used to localize bacteria in food boli and mite bodies. The morphological determination of the mite populations was confirmed by analyses of CO1 and ITS fragment genes.

RESULTS: The following symbiotic bacteria were found in compared mite populations: Wolbachia (two populations), Cardinium (five populations), Bartonella-like (five populations), Blattabacterium-like symbiont (three populations), and Solitalea-like (six populations). From 35 identified OTUs97, only Solitalea was identified in all populations. The next most frequent and abundant sequences were Bacillus, Moraxella, Staphylococcus, Kocuria, and Microbacterium. We suggest that some bacterial species may occasionally be ingested with food. The bacteriocytes were observed in some individuals in all mite populations. Bacteria were not visualized in food boli by staining, but bacteria were found by histological means in ovaria of Wolbachia-infested populations.

CONCLUSION: The presence of Blattabacterium-like, Cardinium, Wolbachia, and Solitalea-like in the eggs of T. putrescentiae indicates mother to offspring (vertical) transmission. RESULTS of this study indicate that diet and habitats influence not only the ingested bacteria but also the symbiotic bacteria of T. putrescentiae.}, } @article {pmid27461737, year = {2016}, author = {Pietri, JE and DeBruhl, H and Sullivan, W}, title = {The rich somatic life of Wolbachia.}, journal = {MicrobiologyOpen}, volume = {5}, number = {6}, pages = {923-936}, pmid = {27461737}, issn = {2045-8827}, support = {R01 GM104486/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology ; *Disease Transmission, Infectious ; Host-Pathogen Interactions/*physiology ; Nematoda/*microbiology ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is an intracellular endosymbiont infecting most arthropod and some filarial nematode species that is vertically transmitted through the maternal lineage. Due to this primary mechanism of transmission, most studies have focused on Wolbachia interactions with the host germline. However, over the last decade many studies have emerged highlighting the prominence of Wolbachia in somatic tissues, implicating somatic tissue tropism as an important aspect of the life history of this endosymbiont. Here, we review our current understanding of Wolbachia-host interactions at both the cellular and organismal level, with a focus on Wolbachia in somatic tissues.}, } @article {pmid27459519, year = {2016}, author = {Ulrich, JN and Beier, JC and Devine, GJ and Hugo, LE}, title = {Heat Sensitivity of wMel Wolbachia during Aedes aegypti Development.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {7}, pages = {e0004873}, pmid = {27459519}, issn = {1935-2735}, mesh = {Aedes/*growth & development/*microbiology ; Animals ; Bacterial Proteins/genetics/metabolism ; Female ; Hot Temperature ; Humans ; Insect Vectors/*growth & development/*microbiology ; Larva/growth & development/microbiology ; Male ; Wolbachia/genetics/*physiology ; }, abstract = {The wMel strain of Wolbachia bacteria is known to prevent dengue and Zika virus transmission in the mosquito vector Aedes aegypti. Accordingly, the release of wMel-infected A. aegypti in endemic regions has been recommended by the World Health Organization as a potential strategy for controlling dengue and Zika outbreaks. However, the utility of this approach could be limited if high temperatures in the aquatic habitats where A. aegypti develop are detrimental to Wolbachia. We exposed wMel-infected A. aegypti eggs and larvae to fluctuating daily temperatures of 30-40°C for three, five, or seven days during their development. We found that Wolbachia levels in females emerging from heat treatments were significantly lower than in the controls that had developed at 20-30°C. Notably, seven days of high temperatures starting at the egg stage reduced Wolbachia levels in emerging females to less than 0.1% of the wMel control levels. However, after adult females returned to 20-30°C for 4-7 days, they experienced differing degrees of Wolbachia recovery. Our findings suggest that the spread of Wolbachia in wild A. aegypti populations and any consequent protection from dengue and Zika viruses might be limited in ecosystems that experience periods of extreme heat, but Wolbachia levels recover partially after temperatures return to normal.}, } @article {pmid27438898, year = {2016}, author = {Mazur, MA and Holecová, M and Lachowska-Cierlik, D and Lis, A and Kubisz, D and Kajtoch, Ł}, title = {Selective sweep of Wolbachia and parthenogenetic host genomes - the example of the weevil Eusomus ovulum.}, journal = {Insect molecular biology}, volume = {25}, number = {6}, pages = {701-711}, doi = {10.1111/imb.12255}, pmid = {27438898}, issn = {1365-2583}, mesh = {Animals ; Cell Nucleus/genetics/metabolism ; DNA, Ribosomal Spacer/genetics/metabolism ; Electron Transport Complex IV/genetics/metabolism ; Female ; *Genes, Bacterial ; *Genetic Variation ; *Genome, Insect ; Insect Proteins/*genetics/metabolism ; *Linkage Disequilibrium ; Mitochondrial Proteins/genetics/metabolism ; Multilocus Sequence Typing ; Parthenogenesis ; Peptide Elongation Factor 1/genetics/metabolism ; Phylogeography ; Sequence Analysis, DNA ; Weevils/*genetics/microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Most parthenogenetic weevil species are postulated to have originated via hybridization, but Wolbachia has also been speculated to play a role via the induction of parthenogenesis. Here, we examine the molecular diversity of Wolbachia and parthenogenetic host genomes. The host species studied here, Eusomus ovulum, is known to be exclusively parthenogenetic and triploid. The E. ovulum populations that we examined had a low genetic diversity of mitochondrial (cytochrome oxidase I gene) and nuclear markers (internal transcribed spacer 2 and elongation factor 1-α gene), and they all were infected by only single bacteria strains (genotyped for five genes according to the multilocus sequence typing system). We found significant signs of linkage disequilibrium and a lack of recombination amongst all of the examined genomes (bacteria and host), which strongly indicates a selective sweep. The lack of heterozygosity in host nuclear genes, missing bisexual populations and selective sweep between the parthenogenetic host and bacteria genomes suggest that parthenogenesis in this species could have originated as a result of infection rather than hybridization. However, the finding that highly similar Wolbachia strains are also present in other parthenogenetic weevils from the same habitat suggests the opposite scenario: bacteria may have infected the already parthenogenetic lineage and taken advantage of the host's unisexual reproduction.}, } @article {pmid27432829, year = {2016}, author = {Whitaker, N and Berry, TM and Rosenthal, N and Gordon, JE and Gonzalez-Rivera, C and Sheehan, KB and Truchan, HK and VieBrock, L and Newton, IL and Carlyon, JA and Christie, PJ}, title = {Chimeric Coupling Proteins Mediate Transfer of Heterologous Type IV Effectors through the Escherichia coli pKM101-Encoded Conjugation Machine.}, journal = {Journal of bacteriology}, volume = {198}, number = {19}, pages = {2701-2718}, pmid = {27432829}, issn = {1098-5530}, support = {F32 AI114182/AI/NIAID NIH HHS/United States ; R01 GM048746/GM/NIGMS NIH HHS/United States ; T32 AI055449/AI/NIAID NIH HHS/United States ; }, mesh = {Conjugation, Genetic/*physiology ; DNA, Bacterial ; Escherichia coli/genetics/*physiology ; Escherichia coli Proteins/genetics/*metabolism ; Gene Expression Regulation, Bacterial/physiology ; Protein Domains ; *Recombinant Fusion Proteins ; Type IV Secretion Systems/*physiology ; }, abstract = {UNLABELLED: Bacterial type IV secretion systems (T4SSs) are composed of two major subfamilies, conjugation machines dedicated to DNA transfer and effector translocators for protein transfer. We show here that the Escherichia coli pKM101-encoded conjugation system, coupled with chimeric substrate receptors, can be repurposed for transfer of heterologous effector proteins. The chimeric receptors were composed of the N-terminal transmembrane domain of pKM101-encoded TraJ fused to soluble domains of VirD4 homologs functioning in Agrobacterium tumefaciens, Anaplasma phagocytophilum, or Wolbachia pipientis A chimeric receptor assembled from A. tumefaciens VirD4 (VirD4At) mediated transfer of a MOBQ plasmid (pML122) and A. tumefaciens effector proteins (VirE2, VirE3, and VirF) through the pKM101 transfer channel. Equivalent chimeric receptors assembled from the rickettsial VirD4 homologs similarly supported the transfer of known or candidate effectors from rickettsial species. These findings establish a proof of principle for use of the dedicated pKM101 conjugation channel, coupled with chimeric substrate receptors, to screen for translocation competency of protein effectors from recalcitrant species. Many T4SS receptors carry sequence-variable C-terminal domains (CTDs) with unknown function. While VirD4At and the TraJ/VirD4At chimera with their CTDs deleted supported pML122 transfer at wild-type levels, ΔCTD variants supported transfer of protein substrates at strongly diminished or elevated levels. We were unable to detect binding of VirD4At's CTD to the VirE2 effector, although other VirD4At domains bound this substrate in vitro We propose that CTDs evolved to govern the dynamics of substrate presentation to the T4SS either through transient substrate contacts or by controlling substrate access to other receptor domains.

IMPORTANCE: Bacterial type IV secretion systems (T4SSs) display striking versatility in their capacity to translocate DNA and protein substrates to prokaryotic and eukaryotic target cells. A hexameric ATPase, the type IV coupling protein (T4CP), functions as a substrate receptor for nearly all T4SSs. Here, we report that chimeric T4CPs mediate transfer of effector proteins through the Escherichia coli pKM101-encoded conjugation system. Studies with these repurposed conjugation systems established a role for acidic C-terminal domains of T4CPs in regulating substrate translocation. Our findings advance a mechanistic understanding of T4CP receptor activity and, further, support a model in which T4SS channels function as passive conduits for any DNA or protein substrates that successfully engage with and pass through the T4CP specificity checkpoint.}, } @article {pmid27419175, year = {2016}, author = {Dickens, BL and Yang, J and Cook, AR and Carrasco, LR}, title = {Time to Empower Release of Insects Carrying a Dominant Lethal and Wolbachia Against Zika.}, journal = {Open forum infectious diseases}, volume = {3}, number = {2}, pages = {ofw103}, pmid = {27419175}, issn = {2328-8957}, abstract = {RIDL (release of insects with dominant lethality) and Wolbachia are 2 potentially powerful tools in the fight against Zika, but their technological advancement is being hampered by policy barriers. In this study, we discuss what could be done to overcome these regulatory deadlocks.}, } @article {pmid27417081, year = {2016}, author = {Al-Abd, NM and Nor, ZM and Mansor, M and Hasan, MS and Kassim, M}, title = {Antifilarial and Antibiotic Activities of Methanolic Extracts of Melaleuca cajuputi Flowers.}, journal = {The Korean journal of parasitology}, volume = {54}, number = {3}, pages = {273-280}, pmid = {27417081}, issn = {1738-0006}, mesh = {Aedes ; Animals ; Anti-Bacterial Agents/isolation & purification/*pharmacology ; Biological Assay ; Brugia pahangi/*drug effects ; Cell Line ; Female ; Filaricides/isolation & purification/*pharmacology ; Flowers/*chemistry ; Locomotion/drug effects ; Male ; Melaleuca/*chemistry ; Methanol ; Microscopy, Electron ; Plant Extracts/isolation & purification/*pharmacology ; Polymerase Chain Reaction ; Solvents ; Symbiosis/drug effects ; Wolbachia/*drug effects ; }, abstract = {We evaluated the activity of methanolic extracts of Melaleuca cajuputi flowers against the filarial worm Brugia pahangi and its bacterial endosymbiont Wolbachia. Anti-Wolbachia activity was measured in worms and in Aedes albopictus Aa23 cells by PCR, electron microscopy, and other biological assays. In particular, microfilarial release, worm motility, and viability were determined. M. cajuputi flower extracts were found to significantly reduce Wolbachia endosymbionts in Aa23 cells, Wolbachia surface protein, and microfilarial release, as well as the viability and motility of adult worms. Anti-Wolbachia activity was further confirmed by observation of degraded and phagocytized Wolbachia in worms treated with the flower extracts. The data provided in vitro and in vivo evidence that M. cajuputi flower extracts inhibit Wolbachia, an activity that may be exploited as an alternative strategy to treat human lymphatic filariasis.}, } @article {pmid27417080, year = {2016}, author = {Dyab, AK and Galal, LA and Mahmoud, AE and Mokhtar, Y}, title = {Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate.}, journal = {The Korean journal of parasitology}, volume = {54}, number = {3}, pages = {265-272}, pmid = {27417080}, issn = {1738-0006}, mesh = {Animals ; Culicidae/*microbiology/parasitology ; DNA, Bacterial/*analysis/genetics ; Dirofilaria immitis/*microbiology ; Dirofilaria repens/*microbiology ; Egypt ; Female ; Multiplex Polymerase Chain Reaction ; Wolbachia/genetics/*isolation & purification ; Wuchereria bancrofti/*microbiology ; }, abstract = {Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites.}, } @article {pmid27406568, year = {2016}, author = {Paredes, JC and Herren, JK and Schüpfer, F and Lemaitre, B}, title = {The Role of Lipid Competition for Endosymbiont-Mediated Protection against Parasitoid Wasps in Drosophila.}, journal = {mBio}, volume = {7}, number = {4}, pages = {}, pmid = {27406568}, issn = {2150-7511}, mesh = {Animals ; Drosophila melanogaster/metabolism/*microbiology/*parasitology ; Hemolymph/chemistry ; Host-Parasite Interactions ; Hymenoptera/*growth & development/*metabolism ; *Lipid Metabolism ; Lipids/analysis ; Spiroplasma/*growth & development/*metabolism ; Symbiosis ; }, abstract = {UNLABELLED: Insects commonly harbor facultative bacterial endosymbionts, such as Wolbachia and Spiroplasma species, that are vertically transmitted from mothers to their offspring. These endosymbiontic bacteria increase their propagation by manipulating host reproduction or by protecting their hosts against natural enemies. While an increasing number of studies have reported endosymbiont-mediated protection, little is known about the mechanisms underlying this protection. Here, we analyze the mechanisms underlying protection from parasitoid wasps in Drosophila melanogaster mediated by its facultative endosymbiont Spiroplasma poulsonii Our results indicate that S. poulsonii exerts protection against two distantly related wasp species, Leptopilina boulardi and Asobara tabida S. poulsonii-mediated protection against parasitoid wasps takes place at the pupal stage and is not associated with an increased cellular immune response. In this work, we provide three important observations that support the notion that S. poulsonii bacteria and wasp larvae compete for host lipids and that this competition underlies symbiont-mediated protection. First, lipid quantification shows that both S. poulsonii and parasitoid wasps deplete D. melanogaster hemolymph lipids. Second, the depletion of hemolymphatic lipids using the Lpp RNA interference (Lpp RNAi) construct reduces wasp success in larvae that are not infected with S. poulsonii and blocks S. poulsonii growth. Third, we show that the growth of S. poulsonii bacteria is not affected by the presence of the wasps, indicating that when S. poulsonii is present, larval wasps will develop in a lipid-depleted environment. We propose that competition for host lipids may be relevant to endosymbiont-mediated protection in other systems and could explain the broad spectrum of protection provided.

IMPORTANCE: Virtually all insects, including crop pests and disease vectors, harbor facultative bacterial endosymbionts. They are vertically transmitted from mothers to their offspring, and some protect their host against pathogens. Here, we studied the mechanism of protection against parasitoid wasps mediated by the Drosophila melanogaster endosymbiont Spiroplasma poulsonii Using genetic manipulation of the host, we provide strong evidence supporting the hypothesis that competition for host lipids underlies S. poulsonii-mediated protection against parasitoid wasps. We propose that lipid competition-based protection may not be restricted to Spiroplasma bacteria but could also apply other endosymbionts, notably Wolbachia bacteria, which can suppress human disease-causing viruses in insect hosts.}, } @article {pmid27393640, year = {2016}, author = {Hernández-Roldán, JL and Dapporto, L and Dincă, V and Vicente, JC and Hornett, EA and Šíchová, J and Lukhtanov, VA and Talavera, G and Vila, R}, title = {Integrative analyses unveil speciation linked to host plant shift in Spialia butterflies.}, journal = {Molecular ecology}, volume = {25}, number = {17}, pages = {4267-4284}, doi = {10.1111/mec.13756}, pmid = {27393640}, issn = {1365-294X}, mesh = {Africa, Northern ; Altitude ; Animals ; Butterflies/*classification ; DNA, Mitochondrial/genetics ; Europe, Eastern ; *Genetic Speciation ; Larva ; *Phylogeny ; Wolbachia ; }, abstract = {Discovering cryptic species in well-studied areas and taxonomic groups can have profound implications in understanding eco-evolutionary processes and in nature conservation because such groups often involve research models and act as flagship taxa for nature management. In this study, we use an array of techniques to study the butterflies in the Spialia sertorius species group (Lepidoptera, Hesperiidae). The integration of genetic, chemical, cytogenetic, morphological, ecological and microbiological data indicates that the sertorius species complex includes at least five species that differentiated during the last three million years. As a result, we propose the restitution of the species status for two taxa often treated as subspecies, Spialia ali (Oberthür, 1881) stat. rest. and Spialia therapne (Rambur, 1832) stat. rest., and describe a new cryptic species Spialia rosae Hernández-Roldán, Dapporto, Dincă, Vicente & Vila sp. nov. Spialia sertorius (Hoffmannsegg, 1804) and S. rosae are sympatric and synmorphic, but show constant differences in mitochondrial DNA, chemical profiles and ecology, suggesting that S. rosae represents a case of ecological speciation involving larval host plant and altitudinal shift, and apparently associated with Wolbachia infection. This study exemplifies how a multidisciplinary approach can reveal elusive cases of hidden diversity.}, } @article {pmid27381293, year = {2016}, author = {Sheehan, KB and Martin, M and Lesser, CF and Isberg, RR and Newton, IL}, title = {Identification and Characterization of a Candidate Wolbachia pipientis Type IV Effector That Interacts with the Actin Cytoskeleton.}, journal = {mBio}, volume = {7}, number = {4}, pages = {}, pmid = {27381293}, issn = {2150-7511}, support = {R01 AI064285/AI/NIAID NIH HHS/United States ; }, mesh = {Actin Cytoskeleton/*metabolism ; Animals ; Bacterial Proteins/*genetics/*metabolism ; Drosophila melanogaster/*microbiology ; Protein Binding ; Protein Interaction Mapping ; Recombinant Proteins/genetics/metabolism ; Saccharomyces cerevisiae/genetics/growth & development ; Wolbachia/*genetics/*metabolism ; }, abstract = {UNLABELLED: Many bacteria live as intracellular symbionts, causing persistent infections within insects. One extraordinarily common infection is that of Wolbachia pipientis, which infects 40% of insect species and induces reproductive effects. The bacteria are passed from generation to generation both vertically (through the oocyte) and horizontally (by environmental transmission). Maintenance of the infection within Drosophila melanogaster is sensitive to the regulation of actin, as Wolbachia inefficiently colonizes strains hemizygous for the profilin or villin genes. Therefore, we hypothesized that Wolbachia must depend on the host actin cytoskeleton. In this study, we identify and characterize a Wolbachia protein (WD0830) that is predicted to be secreted by the bacterial parasite. Expression of WD0830 in a model eukaryote (the yeast Saccharomyces cerevisiae) induces a growth defect associated with the appearance of aberrant, filamentous structures which colocalize with rhodamine-phalloidin-stained actin. Purified WD0830 bundles actin in vitro and cosediments with actin filaments, suggesting a direct interaction of the two proteins. We characterized the expression of WD0830 throughout Drosophila development and found it to be upregulated in third-instar larvae, peaking in early pupation, during the critical formation of adult tissues, including the reproductive system. In transgenic flies, heterologously expressed WD0830 localizes to the developing oocyte. Additionally, overexpression of WD0830 results in increased Wolbachia titers in whole flies, in stage 9 and 10 oocytes, and in embryos, compared to controls, suggesting that the protein may facilitate Wolbachia's replication or transmission. Therefore, this candidate secreted effector may play a role in Wolbachia's infection of and persistence within host niches.

IMPORTANCE: The obligate intracellular Wolbachia pipientis is a ubiquitous alphaproteobacterial symbiont of arthropods and nematodes and is related to the rickettsial pathogens Ehrlichia spp. and Anaplasma spp. Studies of Wolbachia cell biology suggest that this bacterium relies on host actin for efficient proliferation and transmission between generations. Here, we identified and characterized a Wolbachia protein that localizes to and manipulates the eukaryotic actin cytoskeleton, is expressed by Wolbachia during host development, and alters Wolbachia titers and localization in transgenic fruit flies. We hypothesize that WD0830 may be utilized by the bacterium to facilitate replication in or invasion of different niches during host development.}, } @article {pmid27364935, year = {2016}, author = {Aliota, MT and Peinado, SA and Velez, ID and Osorio, JE}, title = {The wMel strain of Wolbachia Reduces Transmission of Zika virus by Aedes aegypti.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {28792}, pmid = {27364935}, issn = {2045-2322}, support = {R21 AI117413/AI/NIAID NIH HHS/United States ; T32 AI007414/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/virology ; Animals ; Chlorocebus aethiops ; Host-Pathogen Interactions ; Humans ; Insect Vectors/*microbiology/virology ; Pest Control, Biological/*methods ; Vero Cells ; Wolbachia/*physiology ; Zika Virus/physiology ; Zika Virus Infection/*prevention & control/transmission/virology ; }, abstract = {Zika virus (ZIKV) is causing an explosive outbreak of febrile disease in the Americas. There are no effective antiviral therapies or licensed vaccines for this virus, and mosquito control strategies have not been adequate to contain the virus. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against other arboviruses. At present, it is unknown whether or not ZIKV can infect, disseminate, and be transmitted by Wolbachia-infected Ae. aegypti. Using Ae. aegypti infected with the wMel strain of Wolbachia that are being released in Medellin, Colombia, we report that these mosquitoes have reduced vector competence for ZIKV. These results support the use of Wolbachia biocontrol as a multivalent strategy against Ae. aegypti-transmitted viruses.}, } @article {pmid27362709, year = {2016}, author = {Sikulu-Lord, MT and Maia, MF and Milali, MP and Henry, M and Mkandawile, G and Kho, EA and Wirtz, RA and Hugo, LE and Dowell, FE and Devine, GJ}, title = {Rapid and Non-destructive Detection and Identification of Two Strains of Wolbachia in Aedes aegypti by Near-Infrared Spectroscopy.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {6}, pages = {e0004759}, pmid = {27362709}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Female ; Host-Parasite Interactions ; Male ; Mosquito Control ; Mosquito Vectors/*microbiology ; Regression Analysis ; Spectroscopy, Near-Infrared/*methods ; Time Factors ; Wolbachia/*classification/*isolation & purification/physiology ; }, abstract = {The release of Wolbachia infected mosquitoes is likely to form a key component of disease control strategies in the near future. We investigated the potential of using near-infrared spectroscopy (NIRS) to simultaneously detect and identify two strains of Wolbachia pipientis (wMelPop and wMel) in male and female laboratory-reared Aedes aegypti mosquitoes. Our aim is to find faster, cheaper alternatives for monitoring those releases than the molecular diagnostic techniques that are currently in use. Our findings indicate that NIRS can differentiate females and males infected with wMelPop from uninfected wild type samples with an accuracy of 96% (N = 299) and 87.5% (N = 377), respectively. Similarly, females and males infected with wMel were differentiated from uninfected wild type samples with accuracies of 92% (N = 352) and 89% (N = 444). NIRS could differentiate wMelPop and wMel transinfected females with an accuracy of 96.6% (N = 442) and males with an accuracy of 84.5% (N = 443). This non-destructive technique is faster than the standard polymerase chain reaction diagnostic techniques. After the purchase of a NIRS spectrometer, the technique requires little sample processing and does not consume any reagents.}, } @article {pmid27353950, year = {2017}, author = {Strunov, A and Schneider, DI and Albertson, R and Miller, WJ}, title = {Restricted distribution and lateralization of mutualistic Wolbachia in the Drosophila brain.}, journal = {Cellular microbiology}, volume = {19}, number = {1}, pages = {}, doi = {10.1111/cmi.12639}, pmid = {27353950}, issn = {1462-5822}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Brain/*microbiology ; Drosophila/*microbiology/physiology ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Sexual Behavior, Animal ; *Symbiosis ; Wolbachia/*isolation & purification/*physiology ; }, abstract = {Microbial symbionts are universal entities of all living organisms that can significantly affect host fitness traits in manifold ways but, even more fascinating, also their behaviour. Although better known from parasitic symbionts, we currently lack any cases where 'neurotrophic' symbionts have co-evolved mutualistic behavioural interactions from which both partners profit. By theory, most mutualistic associations have originated from ancestral parasitic ones during their long-term co-evolution towards a cost-benefit equilibrium. To manipulate host behaviour in a way where both partners benefit in a reciprocal manner, the symbiont has to target and remain restricted to defined host brain regions to minimize unnecessary fitness costs. By using the classic Drosophila paulistorum model system we demonstrate that (i) mutualistic Wolbachia are restricted to various Drosophila brain areas, (ii) form bacteriocyte-like structures within the brain, (iii) exhibit strictly lateral tropism, and (iv) finally propose that their selective neuronal infection affects host sexual behaviour adaptively.}, } @article {pmid27342560, year = {2016}, author = {Christensen, S and Pérez Dulzaides, R and Hedrick, VE and Momtaz, AJ and Nakayasu, ES and Paul, LN and Serbus, LR}, title = {Wolbachia Endosymbionts Modify Drosophila Ovary Protein Levels in a Context-Dependent Manner.}, journal = {Applied and environmental microbiology}, volume = {82}, number = {17}, pages = {5354-5363}, pmid = {27342560}, issn = {1098-5336}, support = {T34 GM083688/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila Proteins/genetics/*metabolism ; Drosophila melanogaster/genetics/*microbiology/*physiology ; Female ; Host-Pathogen Interactions ; Ovary/metabolism/microbiology ; Proteomics ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {UNLABELLED: Endosymbiosis is a unique form of interaction between organisms, with one organism dwelling inside the other. One of the most widespread endosymbionts is Wolbachia pipientis, a maternally transmitted bacterium carried by insects, crustaceans, mites, and filarial nematodes. Although candidate proteins that contribute to maternal transmission have been identified, the molecular basis for maternal Wolbachia transmission remains largely unknown. To investigate transmission-related processes in response to Wolbachia infection, ovarian proteomes were analyzed from Wolbachia-infected Drosophila melanogaster and D. simulans. Endogenous and variant host-strain combinations were investigated. Significant and differentially abundant ovarian proteins were detected, indicating substantial regulatory changes in response to Wolbachia Variant Wolbachia strains were associated with a broader impact on the ovary proteome than endogenous Wolbachia strains. The D. melanogaster ovarian environment also exhibited a higher level of diversity of proteomic responses to Wolbachia than D. simulans. Overall, many Wolbachia-responsive ovarian proteins detected in this study were consistent with expectations from the experimental literature. This suggests that context-specific changes in protein abundance contribute to Wolbachia manipulation of transmission-related mechanisms in oogenesis.

IMPORTANCE: Millions of insect species naturally carry bacterial endosymbionts called Wolbachia. Wolbachia bacteria are transmitted by females to their offspring through a robust egg-loading mechanism. The molecular basis for Wolbachia transmission remains poorly understood at this time, however. This proteomic study identified specific fruit fly ovarian proteins as being upregulated or downregulated in response to Wolbachia infection. The majority of these protein responses correlated specifically with the type of host and Wolbachia strain involved. This work corroborates previously identified factors and mechanisms while also framing the broader context of ovarian manipulation by Wolbachia.}, } @article {pmid27335421, year = {2016}, author = {Martinez, J and Cogni, R and Cao, C and Smith, S and Illingworth, CJ and Jiggins, FM}, title = {Addicted? Reduced host resistance in populations with defensive symbionts.}, journal = {Proceedings. Biological sciences}, volume = {283}, number = {1833}, pages = {}, pmid = {27335421}, issn = {1471-2954}, support = {281668/ERC_/European Research Council/International ; 101239/Z/13/Z/WT_/Wellcome Trust/United Kingdom ; WT094664MA/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Alleles ; Animals ; Disease Resistance/*genetics ; Drosophila melanogaster/*microbiology/*virology ; Evolution, Molecular ; *Genes, Insect ; Insect Viruses ; *Selection, Genetic ; *Symbiosis ; Wolbachia ; }, abstract = {Heritable symbionts that protect their hosts from pathogens have been described in a wide range of insect species. By reducing the incidence or severity of infection, these symbionts have the potential to reduce the strength of selection on genes in the insect genome that increase resistance. Therefore, the presence of such symbionts may slow down the evolution of resistance. Here we investigated this idea by exposing Drosophila melanogaster populations to infection with the pathogenic Drosophila C virus (DCV) in the presence or absence of Wolbachia, a heritable symbiont of arthropods that confers protection against viruses. After nine generations of selection, we found that resistance to DCV had increased in all populations. However, in the presence of Wolbachia the resistant allele of pastrel-a gene that has a major effect on resistance to DCV-was at a lower frequency than in the symbiont-free populations. This finding suggests that defensive symbionts have the potential to hamper the evolution of insect resistance genes, potentially leading to a state of evolutionary addiction where the genetically susceptible insect host mostly relies on its symbiont to fight pathogens.}, } @article {pmid27313167, year = {2016}, author = {Suh, E and Fu, Y and Mercer, DR and Dobson, SL}, title = {Interaction of Wolbachia and Bloodmeal Type in Artificially Infected Aedes albopictus (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {53}, number = {5}, pages = {1156-1162}, pmid = {27313167}, issn = {1938-2928}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; }, abstract = {Maternally inherited Wolbachia bacteria are being introduced into vector mosquito populations, with the goal of reducing the transmission of diseases such as dengue fever. The infection dynamics of Wolbachia depends upon the ability of Wolbachia to manipulate host reproduction as well as any fitness costs imposed upon the host. Some vector mosquito species are opportunistic blood feeders, utilizing both human and nonhuman vertebrate hosts, and the effects of bloodmeal source on Wolbachia phenotype is not well understood. Here we transfer wMelPop Wolbachia from Drosophila melanogaster (Meigen) into wild-type Aedes albopictus (Skuse) and characterize the resulting triple infection by examining for an effect of human and mouse blood on the Wolbachia infection persistence and phenotypes. When provided with human blood, the triple Wolbachia infection was persistent, with high maternal inheritance and relatively little fecundity cost, and a pattern of imperfect unidirectional cytoplasmic incompatibility was observed in mating experiments between wild-type and triply infected individuals. With mouse blood, reduced female fecundity and low maternal inheritance were observed in wMelPop-infected females, which affected the typical pattern of unidirectional CI. Our findings indicate the interactive effects of Wolbachia infection and blood source drive distinct shifts in the Wolbachia-host symbiotic association.}, } @article {pmid27308293, year = {2016}, author = {Karami, M and Moosa-Kazemi, SH and Oshaghi, MA and Vatandoost, H and Sedaghat, MM and Rajabnia, R and Hosseini, M and Maleki-Ravasan, N and Yahyapour, Y and Ferdosi-Shahandashti, E}, title = {Wolbachia Endobacteria in Natural Populations of Culex pipiens of Iran and Its Phylogenetic Congruence.}, journal = {Journal of arthropod-borne diseases}, volume = {10}, number = {3}, pages = {347-363}, pmid = {27308293}, issn = {2322-1984}, abstract = {BACKGROUND: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mosquitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases.

METHODS: Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these endosymbiont bacteria in Culex pipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene.

RESULTS: Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phylogenetic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world.

CONCLUSION: It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evolutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population reproductive incompatibility pattern and its usefulness as a bio-agent control measure.}, } @article {pmid27296468, year = {2016}, author = {Chen, YT and Zhang, YK and Du, WX and Jin, PY and Hong, XY}, title = {Geography has a greater effect than Wolbachia infection on population genetic structure in the spider mite, Tetranychus pueraricola.}, journal = {Bulletin of entomological research}, volume = {106}, number = {5}, pages = {685-694}, doi = {10.1017/S0007485316000444}, pmid = {27296468}, issn = {1475-2670}, mesh = {Animals ; DNA, Mitochondrial/chemistry ; *Genetic Variation ; Geography ; Haplotypes ; Multilocus Sequence Typing ; Phylogeny ; Phylogeography ; Sequence Analysis, DNA ; Social Isolation ; Tetranychidae/*genetics/microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an intracellular symbiotic bacterium that infects various spider mite species and is associated with alterations in host reproduction, which indicates the potential role in mite evolution. However, studies of Wolbachia infections in the spider mite Tetranychus pueraricola, a major agricultural pest, are limited. Here, we used multilocus sequence typing to determine Wolbachia infection status and examined the relationship between Wolbachia infection status and mitochondrial diversity in T. pueraricola from 12 populations in China. The prevalence of Wolbachia ranged from 2.8 to 50%, and three strains (wTpue1, wTpue2, and wTpue3) were identified. We also found double infections (wTpue1 + wTpue3) within the same individuals. Furthermore, the wTpue1 strain caused weak cytoplasmic incompatibility (CI) (egg hatchability ~55%), whereas another widespread strain, wTpue3, did not induce CI. There was no reduction in mitochondrial DNA (mtDNA) or nuclear DNA diversity among infected individuals, and mtDNA haplotypes did not correspond to specific Wolbachia strains. Phylogenetic analysis and analysis of molecular variance revealed that the distribution of mtDNA and nuclear DNA haplotypes were significantly associated with geography. These findings indicate that Wolbachia infection in T. pueraricola is complex, but T. pueraricola genetic differentiation likely resulted from substantial geographic isolation.}, } @article {pmid27291078, year = {2016}, author = {Zhang, YK and Chen, YT and Yang, K and Qiao, GX and Hong, XY}, title = {Screening of spider mites (Acari: Tetranychidae) for reproductive endosymbionts reveals links between co-infection and evolutionary history.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {27900}, pmid = {27291078}, issn = {2045-2322}, mesh = {Animals ; Bacteroidetes/classification/genetics/*physiology ; Biological Evolution ; Coinfection ; Electron Transport Complex IV/chemistry/genetics ; Host Specificity ; Phylogeny ; RNA, Ribosomal, 18S/chemistry/genetics ; RNA, Ribosomal, 28S/chemistry/genetics ; Reproduction ; Rickettsia/classification/genetics/*physiology ; Sequence Analysis, DNA ; Spiroplasma/classification/genetics/*physiology ; Symbiosis ; Tetranychidae/classification/genetics/*microbiology ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Reproductive endosymbionts have been shown to have wide-ranging effects on many aspects of their hosts' biology. A first step to understanding how these endosymbionts interact with their hosts is to determine their incidences. Here, we screened for four reproductive endosymbionts (Wolbachia, Cardinium, Spiroplasma and Rickettsia) in 28 populations of spider mites (Acari: Tetranychidae) representing 12 species. Each of the four endosymbionts were identified in at least some of the tested specimens, and their infection patterns showed variations at the species-level and population-level, suggesting their distributions can be correlated with both the phylogeny and ecology of the hosts. Co-infections of unrelated bacteria, especially double infections of Wolbachia and Cardinium within the same individuals were common. Spiroplasma and Rickettsia infections were specific to particular host species, respectively. Further, the evolutionary histories of these endosymbionts were inferred by comparing the phylogenies of them and their hosts. These findings can help to clarify the interactions between endosymbionts and arthropods.}, } @article {pmid27282489, year = {2016}, author = {Richardson, KM and Schiffer, M and Griffin, PC and Lee, SF and Hoffmann, AA}, title = {Tropical Drosophila pandora carry Wolbachia infections causing cytoplasmic incompatibility or male killing.}, journal = {Evolution; international journal of organic evolution}, volume = {70}, number = {8}, pages = {1791-1802}, pmid = {27282489}, issn = {1558-5646}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/physiology ; Drosophila/*microbiology/*physiology ; Female ; Male ; Queensland ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Wolbachia infections have been described in several Drosophila species, but relatively few have been assessed for phenotypic effects. Cytoplasmic incompatibility (CI) is the most common phenotypic effect that has been detected, while some infections cause male killing or feminization, and many Wolbachia infections have few host effects. Here, we describe two new infections in a recently described species, Drosophila pandora, one of which causes near-complete CI and near-perfect maternal transmission (the "CI" strain). The other infection is a male killer (the "MK" strain), which we confirm by observing reinitiation of male production following tetracycline treatment. No incompatibility was detected in crosses between CI strain males and MK strain females, and rare MK males do not cause CI. Molecular analyses indicate that the CI and MK infections are distantly related and the CI infection is closely related to the wRi infection of Drosophila simulans. Two population surveys indicate that all individuals are infected with Wolbachia, but the MK infection is uncommon. Given patterns of incompatibility among the strains, the infection dynamics is expected to be governed by the relative fitness of the females, suggesting that the CI infection should have a higher fitness. This was evidenced by changes in infection frequencies and sex ratios in population cages initiated at different starting frequencies of the infections.}, } @article {pmid27279628, year = {2016}, author = {Yañez, O and Gauthier, L and Chantawannakul, P and Neumann, P}, title = {Endosymbiotic bacteria in honey bees: Arsenophonus spp. are not transmitted transovarially.}, journal = {FEMS microbiology letters}, volume = {363}, number = {14}, pages = {}, pmid = {27279628}, issn = {1574-6968}, mesh = {Animal Diseases/microbiology/transmission ; Animals ; *Bacteria/classification/genetics ; Bees/*microbiology ; Enterobacteriaceae/classification/genetics ; Phylogeny ; RNA, Bacterial ; RNA, Ribosomal, 16S ; *Symbiosis ; }, abstract = {Intracellular endosymbiotic bacteria are common and can play a crucial role for insect pathology. Therefore, such bacteria could be a potential key to our understanding of major losses of Western honey bees (Apis mellifera) colonies. However, the transmission and potential effects of endosymbiotic bacteria in A. mellifera and other Apis spp. are poorly understood. Here, we explore the prevalence and transmission of the genera Arsenophonus, Wolbachia, Spiroplasma and Rickettsia in Apis spp. Colonies of A. mellifera (N = 33, with 20 eggs from worker brood cells and 100 adult workers each) as well as mated honey bee queens of A. cerana, A. dorsata and A. florea (N = 12 each) were screened using PCR. While Wolbachia, Spiroplasma and Rickettsia were not detected, Arsenophonus spp. were found in 24.2% of A. mellifera colonies and respective queens as well as in queens of A. dorsata (8.3%) and A. florea (8.3%), but not in A. cerana The absence of Arsenophonus spp. from reproductive organs of A. mellifera queens and surface-sterilized eggs does not support transovarial vertical transmission. Instead, horizontal transmission is most likely.}, } @article {pmid27279553, year = {2016}, author = {DE Pinho Mixão, V and Mendes, AM and Maurício, IL and Calado, MM and Novo, MT and Belo, S and Almeida, AP}, title = {Molecular detection of Wolbachia pipientis in natural populations of mosquito vectors of Dirofilaria immitis from continental Portugal: first detection in Culex theileri.}, journal = {Medical and veterinary entomology}, volume = {30}, number = {3}, pages = {301-309}, doi = {10.1111/mve.12179}, pmid = {27279553}, issn = {1365-2915}, mesh = {Animals ; Culex/*microbiology/parasitology ; DNA, Bacterial/*genetics ; Dirofilaria immitis/isolation & purification ; Dirofilariasis/parasitology/transmission ; Haplotypes ; Mosquito Vectors/*microbiology/parasitology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Portugal ; RNA, Ribosomal, 16S/*genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia pipientis (Rickettsiales: Rickettsiaceae) protects mosquitoes from infections with arboviruses and parasites. However, the effect of its co-infection on vector competence for Dirofilaria immitis (Spirurida: Onchocercidae) in the wild has not been investigated. This study aimed to screen vectors of D. immitis for wPip, to characterize these, and to investigate a possible association between the occurrence of W. pipientis and that of the nematode. The presence of W. pipientis was assessed in the five mosquito potential vectors of D. immitis in Portugal. Polymerase chain reaction (PCR) products were sequenced, and wPip haplotypes were determined by PCR-restricted fragment length polymorphism (RFLP). Results showed that wPip was detected in 61.5% of Culex pipiens (Diptera: Culicidae) pools and 6.3% of Culex theileri pools. wPip 16s rRNA sequences found in Cx. theileri exactly match those from Cx. pipiens, confirming a mosquito origin, rather than a nematode origin, as some specimens were infected with D. immitis. Only wPip haplotype I was found. No association was found between the presence of wPip and D. immitis in mosquitoes and hence a role for this endosymbiont in influencing vectorial competence is yet to be identified. This study contributes to understanding of wPip distribution in mosquito populations and, to the best of the authors' knowledge, is the first report of natural infections by wPip in Cx. theileri.}, } @article {pmid27270667, year = {2016}, author = {Perotti, MA and Young, DK and Braig, HR}, title = {The ghost sex-life of the paedogenetic beetle Micromalthus debilis.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {27364}, pmid = {27270667}, issn = {2045-2322}, support = {BB/N001443/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Coleoptera/*physiology/*radiation effects ; *Hot Temperature ; Parthenogenesis/radiation effects ; Rickettsia/growth & development ; Wolbachia/growth & development ; }, abstract = {Genetic and sexual systems can be evolutionarily dynamic within and among clades. However, identifying the processes responsible for switches between, for instance, sexual and asexual reproduction, or cyclic and non-cyclic life histories remains challenging. When animals evolve parthenogenetic reproduction, information about the sexual mating system becomes lost. Here we report an extraordinary case where we have been able to resurrect sexual adults in a species of beetle that reproduces by parthenogenetic paedogenesis, without the production of adults. Via heat treatment, we were able to artificially induce adult beetles of Micromalthus debilis in order to describe its pre-paedogenetic mating system. Adults showed a highly female biased sex ratio, out-breeding behaviour, and sex-role reversal. Paedogenetic larvae of Micromalthus are infected with the endosymbiotic bacteria Rickettsia and Wolbachia. Clear signs of vestigialization in adults are concurrent with the loss of adults. Our data suggest an ancient female sex ratio bias that predates the loss of adults, perhaps associated with endosymbionts. We propose a model for the transition from a haplodiploid cyclical parthenogenetic life history to parthenogenetic paedogenesis. Paedogenetic development induces a new mechanism of sex ratio bias in midges, wasps and beetles.}, } @article {pmid27270383, year = {2016}, author = {Starkey, LA and Newton, K and Brunker, J and Crowdis, K and Edourad, EJP and Meneus, P and Little, SE}, title = {Prevalence of vector-borne pathogens in dogs from Haiti.}, journal = {Veterinary parasitology}, volume = {224}, number = {}, pages = {7-12}, doi = {10.1016/j.vetpar.2016.04.017}, pmid = {27270383}, issn = {1873-2550}, mesh = {Animals ; Antibodies, Bacterial/blood ; Antibodies, Helminth/blood ; Bacteria/genetics ; Coinfection/epidemiology/veterinary ; Dog Diseases/*epidemiology/microbiology/parasitology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Haiti/epidemiology ; Parasites/genetics ; Polymerase Chain Reaction ; Prevalence ; Tick-Borne Diseases/epidemiology/microbiology/parasitology/*veterinary ; Ticks/microbiology/parasitology ; }, abstract = {Canine vector-borne pathogens are common on some Caribbean islands, but survey data in Haiti are lacking. To determine the prevalence of selected vector-borne pathogens in dogs from Haiti, we tested blood samples collected from 210 owned dogs, 28 (13.3%) of which were infested with Rhipicephalus sanguineus ticks at the time of blood collection. No other tick species were identified on these dogs. A commercially available ELISA identified antibodies to Ehrlichia spp. in 69 (32.9%), antibodies to Anaplasma spp. in 37 (17.6%), and antigen of Dirofilaria immitis in 55 (26.2%); antibodies to Borrelia burgdorferi were not detected in any sample. Molecular assays of whole blood from 207 of the dogs confirmed infection with Ehrlichia canis (15; 7.2%), Anaplasma platys (13; 6.3%), D. immitis (46; 22.2%), Wolbachia spp. (45; 21.7%), Babesia vogeli (16; 7.7%), and Hepatozoon canis (40; 19.3%), but Anaplasma phagocytophilum, Babesia canis, Babesia rossi, Babesia gibsoni, Ehrlichia chaffeensis, Ehrlichia ewingii, or Hepatozoon americanum were not detected. Co-infection with two or more vector-borne pathogens was detected by serology in 42 (20.0%) dogs and by molecular assays in 22 (10.6%) dogs; one dog was co-infected with B. vogeli and E. canis as detected by PCR with D. immitis detected by serology (antigen). Overall, evidence of past or current infection with at least one vector-borne pathogen was identified in 142/210 (67.6%) dogs in this study, underscoring the common nature of these pathogens, some of which are zoonotic, in Haiti.}, } @article {pmid27267358, year = {2016}, author = {Kang, JG and Won, S and Kim, HW and Kim, BJ and Park, BK and Park, TS and Seo, HY and Chae, JS}, title = {Molecular detection of Bartonella spp. in terrestrial leeches (Haemadipsa rjukjuana) feeding on human and animal blood in Gageo-do, Republic of Korea.}, journal = {Parasites & vectors}, volume = {9}, number = {1}, pages = {326}, pmid = {27267358}, issn = {1756-3305}, mesh = {Animals ; Bartonella/genetics/*isolation & purification ; Birds/blood ; DNA, Bacterial/genetics ; DNA, Intergenic/genetics ; Feeding Behavior ; Genotype ; Humans ; Islands ; Leeches/*microbiology ; Mice ; Mustelidae/blood ; Polymerase Chain Reaction ; Republic of Korea ; }, abstract = {BACKGROUND: Leeches can transmit pathogens and are therefore potentially hazardous to human and animal health. However, only a few studies of diseases transmitted by land leeches have been reported. The purpose of the present study was to analyse which pathogens are carried in Haemadipsa rjukjuana, the first recorded sanguivorous land leech in the Republic of Korea (ROK).

FINDINGS: A total of 173 H. rjukjuana were collected from Mt. Dock-Sil on Gageo-do Island, ROK during July 2011. Conventional PCR was conducted for analysis of the origin of blood meal, as well as for detection of species of Anaplasma, Bartonella, Borrelia, Ehrlichia, Rickettsia, and Wolbachia in the leech specimens. Bartonella DNA was detected in eight of the specimens studied based on partial ITS sequence analysis. Seven of the DNA samples were closely related to Bartonella grahamii (99.6-100 % similarity), and one sample exhibited a 90.6 % similarity with Bartonella sp. from Taiwan. Sequences of the mitochondrial cytochrome b gene were generated for a total of 35 of the 173 leech internal organ samples. These included sequences of human (n = 10), mouse (n = 8), weasel (n = 6) and bird (n = 11) origin. Of these 35 sequences, 68.5 % were from mammals, including humans, and 31.4 % were from migratory birds that pass through Gageo-do, ROK.

CONCLUSIONS: Although the present study does not provide evidence that leeches indeed transmit Bartonella species to hosts directly, to our knowledge this is the first report on Bartonella DNA being detected from leeches. Therefore, further studies are needed to explore the possibility of zoonotic pathogen transmission by land leeches.}, } @article {pmid27258532, year = {2016}, author = {Bili, M and Cortesero, AM and Mougel, C and Gauthier, JP and Ermel, G and Simon, JC and Outreman, Y and Terrat, S and Mahéo, F and Poinsot, D}, title = {Bacterial Community Diversity Harboured by Interacting Species.}, journal = {PloS one}, volume = {11}, number = {6}, pages = {e0155392}, pmid = {27258532}, issn = {1932-6203}, mesh = {Animals ; Biodiversity ; Coleoptera/*microbiology ; Diptera/*microbiology/parasitology ; Microbiota ; }, abstract = {All animals are infected by microbial partners that can be passengers or residents and influence many biological traits of their hosts. Even if important factors that structure the composition and abundance of microbial communities within and among host individuals have been recently described, such as diet, developmental stage or phylogeny, few studies have conducted cross-taxonomic comparisons, especially on host species related by trophic relationships. Here, we describe and compare the microbial communities associated with the cabbage root fly Delia radicum and its three major parasitoids: the two staphylinid beetles Aleochara bilineata and A. bipustulata and the hymenopteran parasitoid Trybliographa rapae. For each species, two populations from Western France were sampled and microbial communities were described through culture independent methods (454 pyrosequencing). Each sample harbored at least 59 to 261 different bacterial phylotypes but was strongly dominated by one or two. Microbial communities differed markedly in terms of composition and abundance, being mainly influenced by phylogenetic proximity but also geography to a minor extent. Surprisingly, despite their strong trophic interaction, parasitoids shared a very low proportion of microbial partners with their insect host. Three vertically transmitted symbionts from the genus Wolbachia, Rickettsia, and Spiroplasma were found in this study. Among them, Wolbachia and Spiroplasma were found in both the cabbage fly and at least one of its parasitoids, which could result from horizontal transfers through trophic interactions. Phylogenetic analysis showed that this hypothesis may explain some but not all cases. More work is needed to understand the dynamics of symbiotic associations within trophic network and the effect of these bacterial communities on the fitness of their hosts.}, } @article {pmid27243367, year = {2016}, author = {Shaw, WR and Marcenac, P and Childs, LM and Buckee, CO and Baldini, F and Sawadogo, SP and Dabiré, RK and Diabaté, A and Catteruccia, F}, title = {Wolbachia infections in natural Anopheles populations affect egg laying and negatively correlate with Plasmodium development.}, journal = {Nature communications}, volume = {7}, number = {}, pages = {11772}, pmid = {27243367}, issn = {2041-1723}, support = {R21 AI117313/AI/NIAID NIH HHS/United States ; U54 GM088558/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Anopheles/microbiology/parasitology/*physiology ; Burkina Faso/epidemiology ; DNA, Bacterial/isolation & purification ; Disease Transmission, Infectious/prevention & control ; Female ; Host-Pathogen Interactions ; Larva/growth & development ; Malaria/epidemiology/*transmission ; Male ; Maternal Inheritance ; Models, Biological ; Mosquito Vectors/microbiology/parasitology/*physiology ; Oviposition/*physiology ; Plasmodium/*growth & development/pathogenicity ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Symbiosis/physiology ; Wolbachia/isolation & purification/*physiology ; }, abstract = {The maternally inherited alpha-proteobacterium Wolbachia has been proposed as a tool to block transmission of devastating mosquito-borne infectious diseases like dengue and malaria. Here we study the reproductive manipulations induced by a recently identified Wolbachia strain that stably infects natural mosquito populations of a major malaria vector, Anopheles coluzzii, in Burkina Faso. We determine that these infections significantly accelerate egg laying but do not induce cytoplasmic incompatibility or sex-ratio distortion, two parasitic reproductive phenotypes that facilitate the spread of other Wolbachia strains within insect hosts. Analysis of 221 blood-fed A. coluzzii females collected from houses shows a negative correlation between the presence of Plasmodium parasites and Wolbachia infection. A mathematical model incorporating these results predicts that infection with these endosymbionts may reduce malaria prevalence in human populations. These data suggest that Wolbachia may be an important player in malaria transmission dynamics in Sub-Saharan Africa.}, } @article {pmid27237947, year = {2016}, author = {Gerth, M}, title = {Classification of Wolbachia (Alphaproteobacteria, Rickettsiales): No evidence for a distinct supergroup in cave spiders.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {43}, number = {}, pages = {378-380}, doi = {10.1016/j.meegid.2016.05.034}, pmid = {27237947}, issn = {1567-7257}, mesh = {Animals ; Caves/*microbiology ; Genes, Bacterial ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Spiders/*microbiology ; Wolbachia/*classification/genetics/isolation & purification ; }, } @article {pmid27233666, year = {2016}, author = {Ahmed, MZ and Breinholt, JW and Kawahara, AY}, title = {Evidence for common horizontal transmission of Wolbachia among butterflies and moths.}, journal = {BMC evolutionary biology}, volume = {16}, number = {1}, pages = {118}, pmid = {27233666}, issn = {1471-2148}, mesh = {Animals ; Biodiversity ; Butterflies/*microbiology ; Gene Regulatory Networks ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Geography ; Likelihood Functions ; Moths/*microbiology ; Phylogeny ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Wolbachia is one of the most widespread bacteria on Earth. Previous research on Wolbachia-host interactions indicates that the bacterium is typically transferred vertically, from mother to offspring, through the egg cytoplasm. Although horizontal transmission of Wolbachia from one species to another is reported to be common in arthropods, limited direct ecological evidence is available. In this study, we examine horizontal transmission of Wolbachia using a multilocus sequence typing (MLST) strains dataset and used Wolbachia and Lepidoptera genomes to search for evidence for lateral gene transfer (LGT) in Lepidoptera, one of the most diverse cosmopolitan insect orders. We constructed a phylogeny of arthropod-associated MLST Wolbachia strains and calibrated the age of Wolbachia strains associated with lepidopteran species.

RESULTS: Our results reveal inter-specific, inter-generic, inter-familial, and inter-ordinal horizontal transmission of Wolbachia strains, without discernible geographic patterns. We found at least seven probable cases of horizontal transmission among 31 species within Lepidoptera and between Lepidoptera and other arthropod hosts. The divergence time analysis revealed that Wolbachia is recently (22.6-4.7 mya, 95 % HPD) introduced in Lepidoptera. Analysis of nine Lepidoptera genomes (Bombyx mori, Danaus plexippus, Heliconius melpomene, Manduca sexta, Melitaea cinxia, Papilio glaucus, P. polytes, P. xuthus and Plutella xylostella) yielded one possible instance of Wolbachia LGT.

CONCLUSIONS: Our results provide evidence of high incidence of identical and multiple strains of Wolbachia among butterflies and moths, adding Lepidoptera to the growing body of evidence for common horizontal transmission of Wolbachia. This study demonstrates interesting dynamics of this remarkable and influential microorganism.}, } @article {pmid27230747, year = {2016}, author = {Maistrenko, OM and Serga, SV and Vaiserman, AM and Kozeretska, IA}, title = {Longevity-modulating effects of symbiosis: insights from Drosophila-Wolbachia interaction.}, journal = {Biogerontology}, volume = {17}, number = {5-6}, pages = {785-803}, doi = {10.1007/s10522-016-9653-9}, pmid = {27230747}, issn = {1573-6768}, mesh = {Aging/*physiology ; Animals ; Bacterial Proteins/metabolism ; Drosophila/*microbiology/*physiology ; Drosophila Proteins/metabolism ; Longevity/*physiology ; Models, Biological ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {Microbial communities are known to significantly affect various fitness components and survival of their insect hosts, including Drosophila. The composition of symbiotic microbiota has been shown to change with the host's aging. It is unclear whether these changes are caused by the aging process or, vice versa, they affect the host's aging and longevity. Recent findings indicate that fitness and lifespan of Drosophila are affected by endosymbiotic bacteria Wolbachia. These effects, however, are inconsistent and have been reported both to extend and shorten longevity. The main molecular pathways underlying the lifespan-modulating effects of Wolbachia remain unclear, however insulin/insulin-like growth factor, immune deficiency, ecdysteroid synthesis and signaling and c-Jun N-terminal kinase pathways as well as heat shock protein synthesis and autophagy have been proposed to play a role. Here we revise the current evidence that elucidates the impact of Wolbachia endosymbionts on the aging processes in Drosophila.}, } @article {pmid27226403, year = {2016}, author = {Armstrong, SD and Xia, D and Bah, GS and Krishna, R and Ngangyung, HF and LaCourse, EJ and McSorley, HJ and Kengne-Ouafo, JA and Chounna-Ndongmo, PW and Wanji, S and Enyong, PA and Taylor, DW and Blaxter, ML and Wastling, JM and Tanya, VN and Makepeace, BL}, title = {Stage-specific Proteomes from Onchocerca ochengi, Sister Species of the Human River Blindness Parasite, Uncover Adaptations to a Nodular Lifestyle.}, journal = {Molecular & cellular proteomics : MCP}, volume = {15}, number = {8}, pages = {2554-2575}, pmid = {27226403}, issn = {1535-9484}, mesh = {Animals ; Cattle ; Disease Models, Animal ; Female ; Gene Expression Regulation, Developmental ; Host-Parasite Interactions ; Humans ; Male ; Onchocerca/metabolism/*physiology ; Onchocerciasis/*parasitology/veterinary ; Phylogeny ; Protein Interaction Maps ; Proteomics/*methods ; Protozoan Proteins/*metabolism ; }, abstract = {Despite 40 years of control efforts, onchocerciasis (river blindness) remains one of the most important neglected tropical diseases, with 17 million people affected. The etiological agent, Onchocerca volvulus, is a filarial nematode with a complex lifecycle involving several distinct stages in the definitive host and blackfly vector. The challenges of obtaining sufficient material have prevented high-throughput studies and the development of novel strategies for disease control and diagnosis. Here, we utilize the closest relative of O. volvulus, the bovine parasite Onchocerca ochengi, to compare stage-specific proteomes and host-parasite interactions within the secretome. We identified a total of 4260 unique O. ochengi proteins from adult males and females, infective larvae, intrauterine microfilariae, and fluid from intradermal nodules. In addition, 135 proteins were detected from the obligate Wolbachia symbiont. Observed protein families that were enriched in all whole body extracts relative to the complete search database included immunoglobulin-domain proteins, whereas redox and detoxification enzymes and proteins involved in intracellular transport displayed stage-specific overrepresentation. Unexpectedly, the larval stages exhibited enrichment for several mitochondrial-related protein families, including members of peptidase family M16 and proteins which mediate mitochondrial fission and fusion. Quantification of proteins across the lifecycle using the Hi-3 approach supported these qualitative analyses. In nodule fluid, we identified 94 O. ochengi secreted proteins, including homologs of transforming growth factor-β and a second member of a novel 6-ShK toxin domain family, which was originally described from a model filarial nematode (Litomosoides sigmodontis). Strikingly, the 498 bovine proteins identified in nodule fluid were strongly dominated by antimicrobial proteins, especially cathelicidins. This first high-throughput analysis of an Onchocerca spp. proteome across the lifecycle highlights its profound complexity and emphasizes the extremely close relationship between O. ochengi and O. volvulus The insights presented here provide new candidates for vaccine development, drug targeting and diagnostic biomarkers.}, } @article {pmid27225098, year = {2016}, author = {Waltz, E}, title = {US reviews plan to infect mosquitoes with bacteria to stop disease.}, journal = {Nature}, volume = {533}, number = {7604}, pages = {450-451}, doi = {10.1038/533450a}, pmid = {27225098}, issn = {1476-4687}, mesh = {Aedes/genetics/immunology/*microbiology/*virology ; Animals ; Dengue/prevention & control/transmission ; Dengue Virus/growth & development/immunology ; Evaluation Studies as Topic ; Female ; Humans ; *Insect Vectors/genetics/immunology/microbiology/virology ; Male ; Population Dynamics ; Reproducibility of Results ; Reproduction ; United States ; United States Environmental Protection Agency ; United States Food and Drug Administration ; Wolbachia/*immunology/*pathogenicity ; Zika Virus/growth & development/immunology ; Zika Virus Infection/*prevention & control/*transmission ; }, } @article {pmid27217229, year = {2016}, author = {Ndii, MZ and Allingham, D and Hickson, RI and Glass, K}, title = {The effect of Wolbachia on dengue outbreaks when dengue is repeatedly introduced.}, journal = {Theoretical population biology}, volume = {111}, number = {}, pages = {9-15}, doi = {10.1016/j.tpb.2016.05.003}, pmid = {27217229}, issn = {1096-0325}, mesh = {Aedes ; Animals ; Dengue/*epidemiology/prevention & control/transmission ; Disease Outbreaks/*prevention & control ; Humans ; *Models, Theoretical ; *Pest Control, Biological ; Wolbachia/*physiology ; }, abstract = {Use of the Wolbachia bacterium is a proposed new strategy to reduce dengue transmission, which results in around 390 million individuals infected annually. In places with strong variations in climatic conditions such as temperature and rainfall, dengue epidemics generally occur only at a certain time of the year. Where dengue is not endemic, the time of year in which imported cases enter the population plays a crucial role in determining the likelihood of outbreak occurrence. We use a mathematical model to study the effects of Wolbachia on dengue transmission dynamics and dengue seasonality. We focus in regions where dengue is not endemic but can spread due to the presence of a dengue vector and the arrival of people with dengue on a regular basis. Our results show that the time-window in which outbreaks can occur is reduced in the presence of Wolbachia-carrying Aedes aegypti mosquitoes by up to six weeks each year. We find that Wolbachia reduces overall case numbers by up to 80%. The strongest effect is obtained when the amplitude of the seasonal forcing is low (0.02-0.30). The benefits of Wolbachia also depend on the transmission rate, with the bacteria most effective at moderate transmission rates ranging between 0.08-0.12. Such rates are consistent with fitted estimates for Cairns, Australia.}, } @article {pmid27199034, year = {2016}, author = {Berasategui, A and Axelsson, K and Nordlander, G and Schmidt, A and Borg-Karlson, AK and Gershenzon, J and Terenius, O and Kaltenpoth, M}, title = {The gut microbiota of the pine weevil is similar across Europe and resembles that of other conifer-feeding beetles.}, journal = {Molecular ecology}, volume = {25}, number = {16}, pages = {4014-4031}, doi = {10.1111/mec.13702}, pmid = {27199034}, issn = {1365-294X}, mesh = {Animals ; Coleoptera/*microbiology ; *Ecosystem ; Enterobacteriaceae/isolation & purification ; Europe ; Firmicutes/isolation & purification ; *Gastrointestinal Microbiome ; Pinus ; Symbiosis ; Weevils/*microbiology ; Wolbachia/isolation & purification ; }, abstract = {The pine weevil (Hylobius abietis, Coleoptera: Curculionidae) is an important pest of conifer seedlings in Europe. Despite its economic importance, little is known about the composition of its gut microbial community and the role it plays in mediating the weevil's ability to utilize conifers as a food source. Here, we characterized the gut bacterial communities of different populations of H. abietis across Europe and compared them to those of other beetles that occupy similar ecological niches. We demonstrate that the microbial community of H. abietis is similar at higher taxonomic levels (family and genus) across locations in Europe, with Wolbachia as the dominant microbe, followed by Enterobacteria and Firmicutes. Despite this similarity, we observed consistent differences between countries and locations, but not sexes. Our meta-analysis demonstrates that the gut bacterial community of the pine weevil is very similar to that of bark beetles that also exploit conifers as a food source. The Enterobacteriaceae symbionts of both host taxa are especially closely related phylogenetically. Conversely, the microbiota of H. abietis is distinct from that of closely related weevils feeding on nonconifer food sources, suggesting that the microbial community of the pine weevil is determined by the environment and may be relevant to host ecology. Furthermore, several H. abietis-associated members of the Enterobacteriaceae family are known to contain genes involved in terpenoid degradation. As such, we hypothesize that the gut microbial community is important for the utilization of conifer seedlings as a food source, either through the detoxification of plant secondary metabolites or through the supplementation of essential nutrients.}, } @article {pmid27198860, year = {2016}, author = {Kawasaki, Y and Schuler, H and Stauffer, C and Lakatos, F and Kajimura, H}, title = {Wolbachia endosymbionts in haplodiploid and diploid scolytine beetles (Coleoptera: Curculionidae: Scolytinae).}, journal = {Environmental microbiology reports}, volume = {8}, number = {5}, pages = {680-688}, doi = {10.1111/1758-2229.12425}, pmid = {27198860}, issn = {1758-2229}, support = {//Institute for Fermentation/ ; I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; 18405012, 20405025 and 26292083 to H.K.//Aid for Scientific Research/ ; J 3527/FWF_/Austrian Science Fund FWF/Austria ; //Showa-Houkoukai (Ito Chube'e) Foundation/ ; J-3527-B22//Austrian Science Fund FWF project/ ; }, abstract = {Haplodiploidy is a sex determination system in which fertilized diploid eggs develop into females and unfertilized haploid eggs develop into males. The evolutionary explanations for this phenomenon include the possibility that haplodiploidy can be reinforced by infection with endosymbiotic bacteria, such as Wolbachia. The subfamily Scolytinae contains species with haplodiploid and diploid sex determination systems. Thus, we studied the association with Wolbachia in 12 diploid and 11 haplodiploid scolytine beetles by analyzing wsp and multilocus sequence typing (MLST) of five loci in this endosymbiont. Wolbachia genotypes were compared with mitochondrial (COI) and nuclear (EF) genotypes in the scolytines. Eight of the 23 scolytine species were infected with Wolbachia, with haplodiploids at significantly higher rates than diploid species. Cloning and sequencing detected multiple infections with up to six Wolbachia strains in individual species. Phylogenetic analyses of wsp and five MLST genes revealed different Wolbachia strains in scolytines. Comparisons between the beetle and Wolbachia phylogenies revealed that closely related beetles were infected with genetically different Wolbachia strains. These results suggest the horizontal transmission of multiple Wolbachia strains between scolytines. We discuss these results in terms of the evolution of different sex determination systems in scolytine beetles.}, } @article {pmid27194801, year = {2016}, author = {Lindsey, AR and Werren, JH and Richards, S and Stouthamer, R}, title = {Comparative Genomics of a Parthenogenesis-Inducing Wolbachia Symbiont.}, journal = {G3 (Bethesda, Md.)}, volume = {6}, number = {7}, pages = {2113-2123}, pmid = {27194801}, issn = {2160-1836}, support = {U54 HG003273/HG/NHGRI NIH HHS/United States ; }, mesh = {Animals ; Ankyrin Repeat ; Bacterial Proteins/*genetics ; Biological Evolution ; Culicidae/microbiology ; Drosophila/microbiology ; Frameshift Mutation ; *Genome, Bacterial ; Genomics ; Host Specificity ; Open Reading Frames ; Parthenogenesis/*genetics ; Phylogeny ; Symbiosis/*genetics ; Wasps/genetics/*microbiology ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia is an intracellular symbiont of invertebrates responsible for inducing a wide variety of phenotypes in its host. These host-Wolbachia relationships span the continuum from reproductive parasitism to obligate mutualism, and provide a unique system to study genomic changes associated with the evolution of symbiosis. We present the genome sequence from a parthenogenesis-inducing Wolbachia strain (wTpre) infecting the minute parasitoid wasp Trichogramma pretiosum The wTpre genome is the most complete parthenogenesis-inducing Wolbachia genome available to date. We used comparative genomics across 16 Wolbachia strains, representing five supergroups, to identify a core Wolbachia genome of 496 sets of orthologous genes. Only 14 of these sets are unique to Wolbachia when compared to other bacteria from the Rickettsiales. We show that the B supergroup of Wolbachia, of which wTpre is a member, contains a significantly higher number of ankyrin repeat-containing genes than other supergroups. In the wTpre genome, there is evidence for truncation of the protein coding sequences in 20% of ORFs, mostly as a result of frameshift mutations. The wTpre strain represents a conversion from cytoplasmic incompatibility to a parthenogenesis-inducing lifestyle, and is required for reproduction in the Trichogramma host it infects. We hypothesize that the large number of coding frame truncations has accompanied the change in reproductive mode of the wTpre strain.}, } @article {pmid27189996, year = {2016}, author = {Newton, IL and Clark, ME and Kent, BN and Bordenstein, SR and Qu, J and Richards, S and Kelkar, YD and Werren, JH}, title = {Comparative Genomics of Two Closely Related Wolbachia with Different Reproductive Effects on Hosts.}, journal = {Genome biology and evolution}, volume = {8}, number = {5}, pages = {1526-1542}, pmid = {27189996}, issn = {1759-6653}, support = {R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; Arthropods/genetics/microbiology/parasitology ; Cytoplasm/genetics ; Drosophila/*genetics/microbiology/parasitology ; Genome, Bacterial ; Genomics ; Host Specificity/*genetics ; Male ; Mutation ; Phylogeny ; Reproduction/genetics ; Selection, Genetic/*genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia pipientis are obligate intracellular bacteria commonly found in many arthropods. They can induce various reproductive alterations in hosts, including cytoplasmic incompatibility, male-killing, feminization, and parthenogenetic development, and can provide host protection against some viruses and other pathogens. Wolbachia differ from many other primary endosymbionts in arthropods because they undergo frequent horizontal transmission between hosts and are well known for an abundance of mobile elements and relatively high recombination rates. Here, we compare the genomes of two closely related Wolbachia (with 0.57% genome-wide synonymous divergence) that differ in their reproductive effects on hosts. wVitA induces a sperm-egg incompatibility (also known as cytoplasmic incompatibility) in the parasitoid insect Nasonia vitripennis, whereas wUni causes parthenogenetic development in a different parasitoid, Muscidifurax uniraptor Although these bacteria are closely related, the genomic comparison reveals rampant rearrangements, protein truncations (particularly in proteins predicted to be secreted), and elevated substitution rates. These changes occur predominantly in the wUni lineage, and may be due in part to adaptations by wUni to a new host environment, or its phenotypic shift to parthenogenesis induction. However, we conclude that the approximately 8-fold elevated synonymous substitution rate in wUni is due to a either an elevated mutation rate or a greater number of generations per year in wUni, which occurs in semitropical host species. We identify a set of genes whose loss or pseudogenization in the wUni lineage implicates them in the phenotypic shift from cytoplasmic incompatibility to parthenogenesis induction. Finally, comparison of these closely related strains allows us to determine the fine-scale mutation patterns in Wolbachia Although Wolbachia are AT rich, mutation probabilities estimated from 4-fold degenerate sites are not AT biased, and predict an equilibrium AT content much less biased than observed (57-50% AT predicted vs. 76% current content at degenerate sites genome wide). The contrast suggests selection for increased AT content within Wolbachia genomes.}, } @article {pmid27183820, year = {2016}, author = {Dutra, HL and Lopes da Silva, V and da Rocha Fernandes, M and Logullo, C and Maciel-de-Freitas, R and Moreira, LA}, title = {The influence of larval competition on Brazilian Wolbachia-infected Aedes aegypti mosquitoes.}, journal = {Parasites & vectors}, volume = {9}, number = {1}, pages = {282}, pmid = {27183820}, issn = {1756-3305}, mesh = {Aedes/growth & development/*microbiology/physiology ; Animals ; Body Size ; Brazil ; Competitive Behavior ; Dengue/*prevention & control/transmission ; Female ; Humans ; Insect Vectors/growth & development/*microbiology/physiology ; Larva ; Male ; Pest Control, Biological/methods ; Phenotype ; Population Density ; Wings, Animal ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: With field releases starting in Brazil, particular interest must be given to understanding how the endosymbiotic bacterium Wolbachia pipientis affects Aedes aegypti mosquitoes with a Brazilian genetic background. Currently, there is limited information on how the bacterium affects phenotypic traits such as larval development rate, metabolic reserves and morphometric parameters in Ae. aegypti. Here, we analyze for the first time, the effect of Wolbachia on these key phenotypes and consider how this might impact the potential of the bacterium as a disease control agent in Brazil.

METHODS: We examined the influence of the wMel strain of Wolbachia in laboratory Ae. aegypti with a Brazilian genetic background, reared under different larval densities. Pupae formation was counted daily to assess differences in development rates. Levels of metabolic reserves and morphometric parameters were assessed in adults resulting from each larval condition.

RESULTS: wMel infection led to more rapid larval development at higher densities for both males and females, with no effect under less crowded conditions in females. Infection also led to reduced body size at both high and low density, but not at intermediate density, although the scale of this difference was maintained regardless of larval density, in comparison to uninfected individuals. Wing shape also varied significantly between infected and uninfected mosquitoes due to larval density. Glycogen levels in uninfected mosquitoes decreased under higher larval density, but were consistently high with Wolbachia infection, regardless of larval density.

CONCLUSIONS: We demonstrate that the wMel Wolbachia strain can positively influence some important host fitness traits, and that this interaction is directly linked to the conditions in which the host is reared. Combined with previously published data, these results suggest that this Wolbachia strain could be successfully used as part of the Eliminate Dengue Program in Brazil.}, } @article {pmid27177475, year = {2016}, author = {Lauzi, S and Maia, JP and Epis, S and Marcos, R and Pereira, C and Luzzago, C and Santos, M and Puente-Payo, P and Giordano, A and Pajoro, M and Sironi, G and Faustino, A}, title = {Molecular detection of Anaplasma platys, Ehrlichia canis, Hepatozoon canis and Rickettsia monacensis in dogs from Maio Island of Cape Verde archipelago.}, journal = {Ticks and tick-borne diseases}, volume = {7}, number = {5}, pages = {964-969}, doi = {10.1016/j.ttbdis.2016.05.001}, pmid = {27177475}, issn = {1877-9603}, mesh = {Anaplasma/genetics/isolation & purification ; Anaplasmosis/epidemiology ; Animals ; Cabo Verde/epidemiology ; Coccidiosis/epidemiology/*veterinary ; Coinfection/epidemiology/microbiology/veterinary ; Dog Diseases/*epidemiology/microbiology/prevention & control/transmission ; Dogs ; Ehrlichia canis/genetics/isolation & purification ; Ehrlichiosis/epidemiology/*veterinary ; Eucoccidiida/genetics/isolation & purification ; Female ; Humans ; Male ; Rickettsia/genetics/isolation & purification ; Rickettsia Infections/epidemiology/*veterinary ; Tick-Borne Diseases/epidemiology/*microbiology/prevention & control/transmission ; Wolbachia/genetics/isolation & purification ; }, abstract = {Tick-borne diseases are emerging worldwide and have an important zoonotic relevance. Dogs play an important role in the epidemiology of several zoonotic tick-borne pathogens acting as sentinels and/or reservoirs. This study focused on the molecular identification of tick-borne pathogens in blood samples of 153 autochthonous asymptomatic dogs in Maio Island, Cape Verde archipelago. Eighty-four (54.9%) dogs were positive for one or more pathogens. Fifty-five (35.9%) dogs were infected with Hepatozoon canis, 53 (34.6%) with Anaplasma platys, five (3.3%) with Ehrlichia canis and Rickettsia monacensis, an emerging human pathogen, was also identified in a single dog (0.7%). The former three pathogens cause important canine tick-borne diseases that are transmitted or potentially transmitted by Rhipicephalus sanguineus s.l., the only hard tick identified in Cape Verde. Furthermore, Wolbachia spp. was amplified from the blood of one dog. None of the dogs were positive for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Midichloria mitochondrii, Bartonella spp., Babesia spp. or Theileria spp. Fifty-four (35.3%) animals showed single infections and 30 (19.6%) co-infections, with A. platys and H. canis co-infection being the most frequent (28 dogs, 18.3%). The frequency of E. canis infection was statistically different among age groups (P=0.017), being higher among dogs older than 4 years compared to younger dogs. Infection by A. platys was also statistically different among age groups (P=0.031), being higher in dogs younger than 2 years compared to older dogs. The statistical analyses showed no significant association of PCR positivity with gender or location. The frequency of tick-borne pathogens detected in dogs in Maio Island, including R. monacensis, highlights the need to improve diagnosis and control in order to prevent the risk of transmission of these pathogens among dogs and humans living in or travelling to this touristic island.}, } @article {pmid27160085, year = {2016}, author = {Maia, C and Altet, L and Serrano, L and Cristóvão, JM and Tabar, MD and Francino, O and Cardoso, L and Campino, L and Roura, X}, title = {Molecular detection of Leishmania infantum, filariae and Wolbachia spp. in dogs from southern Portugal.}, journal = {Parasites & vectors}, volume = {9}, number = {1}, pages = {170}, pmid = {27160085}, issn = {1756-3305}, mesh = {Animals ; Dirofilaria immitis/genetics/isolation & purification ; Dirofilariasis/epidemiology ; Dog Diseases/*epidemiology/microbiology/parasitology ; Dogs ; Female ; Filariasis/epidemiology/parasitology/*veterinary ; Filarioidea/genetics/isolation & purification/microbiology ; Humans ; Leishmania infantum/genetics/*isolation & purification ; Leishmaniasis, Visceral/epidemiology/parasitology/*veterinary ; Male ; Portugal/epidemiology ; Rickettsiaceae Infections/epidemiology/microbiology/*veterinary ; Wolbachia/genetics/*isolation & purification ; Zoonoses ; }, abstract = {BACKGROUND: Leishmaniosis caused by the protozoan Leishmania infantum and dirofilariosis caused by the nematodes Dirofilaria immitis or Dirofilaria repens are vector-borne zoonoses widely present in the Mediterranean basin. In addition, some studies reported that the endosymbiont Wolbachia spp. play a role in the biology and pathogenesis of filarial parasites. The aim of this work was to evaluate the frequency of mono- and co-infections by L. infantum, filariae and Wolbachia spp. and their association with clinical signs in dogs from the south of Portugal. Leishmanial, filarial and Wolbachia spp. DNA were evaluated by specific real-time polymerase chain reaction (qPCR) assays in blood samples from 230 dogs.

FINDINGS: One hundred and thirty-nine (60.4 %) dogs were qPCR-positive for L. infantum and 26 (11.3 %) for filariae (24 for D. immitis only, one D. immitis and for Acanthocheilonema dracunculoides and another one for Acanthocheilonema reconditum only). Wolbachia spp. DNA was amplified from 16 (64.0 %) out of the 25 D. immitis-positive dogs. Nineteen (8.3 %) dogs were co-infected with L. infantum and D. immitis, including the one (0.4 %) A. drancunculoides-positive animal. In dogs without clinical signs consistent with leishmaniosis and/or dirofilariosis, L. infantum prevalence was 69 %, whereas in those dogs with at least one clinical manifestation compatible with any of the two parasitoses prevalence was 42.7 %. Leishmania prevalence was significantly higher in apparently healthy mongrels (77.2 %) and pets (76.9 %) than in defined-breed dogs (including crosses; 58.8 %) and in dogs with an aptitude other than pet (i.e. farm, guard, hunting, shepherd or stray), respectively, whereas in those dogs with at least one clinical sign, the detection of L. infantum DNA was higher in males (53.3 %) and in those dogs not receiving insect repellents (52.8 %).

CONCLUSIONS: The molecular detection of canine vector-borne disease (CVBD) agents, some of which are zoonotic, reinforces the need to implement efficient prophylactic measures, such as insect repellents and macrocyclic lactones (including compliance to administration), in the geographical areas where these agents are distributed, with the view to prevent infection and disease among mammalian hosts including humans.}, } @article {pmid27159727, year = {2017}, author = {Gaikwad, S and Warudkar, A and Shouche, Y}, title = {Efficacy of DNA barcoding for the species identification of spiders from Western Ghats of India.}, journal = {Mitochondrial DNA. Part A, DNA mapping, sequencing, and analysis}, volume = {28}, number = {5}, pages = {638-644}, doi = {10.3109/24701394.2016.1166219}, pmid = {27159727}, issn = {2470-1408}, mesh = {Animals ; DNA Barcoding, Taxonomic/*methods ; Electron Transport Complex IV/genetics ; Genetic Variation ; India ; Phylogeny ; Sequence Analysis, DNA/*methods ; Species Specificity ; Spiders/*classification/enzymology/genetics ; }, abstract = {DNA barcoding has emerged as an additional tool for taxonomy and as an aid to taxonomic impediments. Due to their extensive morphological variation, spiders are taxonomically challenging. Therefore, all over the world, attempts are being made to DNA barcode species of spiders. Till now no attempts were made to DNA barcode Indian spiders despite their rich diversity. We have generated DNA barcodes for 60 species (n = 112) of spiders for the first time from India. Although only 17 species were correctly identified at the species level, DNA barcoding correctly discriminated 99% of the species studied here. We have also found high intraspecies nucleotide divergence in Plexippus paykulli suggesting cryptic diversity that needs to be studied in detail. Our study also showed non-specific amplification of the Cytochrome Oxidase I (COI) gene of endosymbiont bacteria Wolbachia. However, these cases are very rare and could be resolved by the use of modified or group specific primers.}, } @article {pmid27159401, year = {2016}, author = {, }, title = {Correction: Wolbachia Blocks Viral Genome Replication Early in Infection without a Transcriptional Response by the Endosymbiont or Host Small RNA Pathways.}, journal = {PLoS pathogens}, volume = {12}, number = {5}, pages = {e1005639}, pmid = {27159401}, issn = {1553-7374}, abstract = {[This corrects the article DOI: 10.1371/journal.ppat.1005536.].}, } @article {pmid27156023, year = {2016}, author = {Dutra, HL and Rocha, MN and Dias, FB and Mansur, SB and Caragata, EP and Moreira, LA}, title = {Wolbachia Blocks Currently Circulating Zika Virus Isolates in Brazilian Aedes aegypti Mosquitoes.}, journal = {Cell host & microbe}, volume = {19}, number = {6}, pages = {771-774}, pmid = {27156023}, issn = {1934-6069}, mesh = {Aedes/*microbiology/*virology ; Animals ; Antibiosis ; Brazil ; Disease Transmission, Infectious/prevention & control ; Female ; Insect Vectors/microbiology/virology ; Male ; Saliva/microbiology/virology ; Symbiosis ; Wolbachia/*physiology ; Zika Virus/isolation & purification/*physiology ; Zika Virus Infection/microbiology/prevention & control/transmission/virology ; }, abstract = {The recent association of Zika virus with cases of microcephaly has sparked a global health crisis and highlighted the need for mechanisms to combat the Zika vector, Aedes aegypti mosquitoes. Wolbachia pipientis, a bacterial endosymbiont of insect, has recently garnered attention as a mechanism for arbovirus control. Here we report that Aedes aegypti harboring Wolbachia are highly resistant to infection with two currently circulating Zika virus isolates from the recent Brazilian epidemic. Wolbachia-harboring mosquitoes displayed lower viral prevalence and intensity and decreased disseminated infection and, critically, did not carry infectious virus in the saliva, suggesting that viral transmission was blocked. Our data indicate that the use of Wolbachia-harboring mosquitoes could represent an effective mechanism to reduce Zika virus transmission and should be included as part of Zika control strategies.}, } @article {pmid27151484, year = {2016}, author = {Suroowan, S and Mahomoodally, F and Ragoo, L}, title = {Management and Treatment of Dengue and Chikungunya - Natural Products to the Rescue.}, journal = {Combinatorial chemistry & high throughput screening}, volume = {19}, number = {7}, pages = {554-564}, doi = {10.2174/1386207319666160506123401}, pmid = {27151484}, issn = {1875-5402}, mesh = {Aedes/virology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal/therapeutic use ; Antiviral Agents/*chemistry/*pharmacology ; Biological Products/*chemistry ; Chikungunya Fever/*drug therapy/epidemiology/etiology/transmission ; Dengue/*drug therapy/epidemiology/transmission ; Humans ; Mosquito Control ; Mosquito Vectors/virology ; Neglected Diseases/drug therapy ; Plant Extracts/chemistry/pharmacology ; }, abstract = {Neglected tropical diseases (NTDs) flourish mostly in impoverished developing nations of the world. It is estimated that NTDs plague up to 1 billion people every year thereby inducing a massive economic and health burden worldwide. Following explosive outbreaks mostly in Asia, Latin America, Europe and the Indian Ocean, two common NTDs namely, Chikungunya and Dengue both transmitted by an infected mosquito vector principally Aedes aegypti have emerged as a major public health threat. Given the limitations of conventional medicine in specifically targeting the Chikungunya and Dengue virus (CHIKV and DENV), natural products present an interesting avenue to explore in the quest of developing novel anti; mosquito, CHIKV and DENV agents. In this endeavor, a number of plant extracts, isolated phytochemicals, essential oils and seaweeds have shown promising larvicidal and insecticidal activity against some mosquito vectors as well as anti CHIKV and DENV activity invitro. Other natural products that have depicted good potential against these diseases include; the symbiotic bacterial genus Wolbachia which can largely reduce the life span and infectivity of mosquito vectors and the marine Cyanobacterium Trichodesmium erythraeum which has shown anti- CHIKV activity at minimal cytotoxic level. The impetus of modern drug discovery approaches such as high throughput screening, drug repositioning, synthesis and computer-aided drug design will undeniably enhance the process of developing more stable lead molecules from natural products which have shown promising antiviral activity in-vitro.}, } @article {pmid27144810, year = {2016}, author = {Cattel, J and Martinez, J and Jiggins, F and Mouton, L and Gibert, P}, title = {Wolbachia-mediated protection against viruses in the invasive pest Drosophila suzukii.}, journal = {Insect molecular biology}, volume = {25}, number = {5}, pages = {595-603}, doi = {10.1111/imb.12245}, pmid = {27144810}, issn = {1365-2583}, mesh = {Animals ; Dicistroviridae/*physiology ; Drosophila/*microbiology/virology ; Insect Viruses/*physiology ; Introduced Species ; Nodaviridae/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The maternally inherited bacterium Wolbachia is well known for spreading in natural populations by manipulating the reproduction of its arthropod hosts, but can also have mutualist effects that increase host fitness. In mosquitoes and Drosophila some Wolbachia strains can lead to an increase in survival of virus-infected insects, and in most cases this is associated with reduced accumulation of the virus in host tissues. We investigated if the Wolbachia strain wSuz, which naturally infects Drosophila suzukii, is able to confer protection against Drosophila C virus and Flock House virus in different host genetic backgrounds. We found that this strain can increase host survival upon infection with these two viruses. In some cases this effect was associated with lower viral titres, suggesting that it confers resistance to the viruses rather than allowing the flies to tolerate infection. Our results indicate that, in D. suzukii, the antiviral protection provided by Wolbachia is not correlated to its density as found in other Drosophila species. This study demonstrates a phenotypic effect induced by wSuz on its native host which could explain its maintenance in natural populations of D. suzukii.}, } @article {pmid27132518, year = {2016}, author = {Khoo, JJ and Chen, F and Kho, KL and Ahmad Shanizza, AI and Lim, FS and Tan, KK and Chang, LY and AbuBakar, S}, title = {Bacterial community in Haemaphysalis ticks of domesticated animals from the Orang Asli communities in Malaysia.}, journal = {Ticks and tick-borne diseases}, volume = {7}, number = {5}, pages = {929-937}, doi = {10.1016/j.ttbdis.2016.04.013}, pmid = {27132518}, issn = {1877-9603}, mesh = {Anaplasma/genetics/isolation & purification ; Animals ; *Animals, Domestic ; Bacillus/genetics/isolation & purification ; Bacteria/*isolation & purification ; Cats/parasitology ; Chickens/parasitology ; Coxiella/genetics/isolation & purification ; DNA, Bacterial/genetics ; Dogs/parasitology ; Ehrlichia/genetics/isolation & purification ; Ixodidae/*microbiology ; Malaysia/epidemiology ; *Microbiota ; Mycobacterium/genetics/isolation & purification ; Phylogeny ; Polymerase Chain Reaction ; Pseudomonas/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/isolation & purification ; Sphingomonas/genetics/isolation & purification ; Tick Infestations/epidemiology/*veterinary ; }, abstract = {Ticks are vectors in the transmission of many important infectious diseases in human and animals. Ticks can be readily found in the semi-forested areas such as the settlements of the indigenous people in Malaysia, the Orang Asli. There is still minimal information available on the bacterial agents associated with ticks found in Malaysia. We performed a survey of the bacterial communities associated with ticks collected from domestic animals found in two Orang Asli villages in Malaysia. We collected 62 ticks, microscopically and molecularly identified as related to Haemaphysalis wellingtoni, Haemaphysalis hystricis and Haemaphysalis bispinosa. Bacterial 16s rRNA hypervariable region (V6) amplicon libraries prepared from the tick samples were sequenced on the Ion Torrent PGM platform. We detected a total of 392 possible bacterial genera after pooling and sequencing 20 samples, indicating a diverse bacterial community profile. Dominant taxa include the potential tick endosymbiont, Coxiella. Other dominant taxa include the tick-associated pathogen, Rickettsia, and environmental bacteria such as Bacillus, Mycobacterium, Sphingomonas and Pseudomonas. Other known tick-associated bacteria were also detected, including Anaplasma, Ehrlichia, Rickettsiella and Wolbachia, albeit at very low abundance. Specific PCR was performed on selected samples to identify Rickettsia and Coxiella. Sequence of Rickettsia felis, which causes spotted fever in human and cats, was identified in one sample. Coxiella endosymbionts were detected in three samples. This study provides the baseline knowledge of the microbiome of ticks in Malaysia, focusing on tick-associated bacteria affecting the Orang Asli communities. The role of the herein found Coxiella and Rickettsia in tick physiology or disease transmission merits further investigation.}, } @article {pmid27126029, year = {2016}, author = {Dobson, SL and Bordenstein, SR and Rose, RI}, title = {Wolbachia mosquito control: Regulated.}, journal = {Science (New York, N.Y.)}, volume = {352}, number = {6285}, pages = {526-527}, doi = {10.1126/science.352.6285.526-b}, pmid = {27126029}, issn = {1095-9203}, mesh = {Animals ; Culicidae/*microbiology ; Dengue/*prevention & control ; Mosquito Control/*methods ; *Wolbachia ; }, } @article {pmid29491905, year = {2016}, author = {Humphreys, DP and Rundle, HD and Dyer, KA}, title = {Patterns of reproductive isolation in the Drosophila subquinaria complex: can reinforced premating isolation cascade to other species?.}, journal = {Current zoology}, volume = {62}, number = {2}, pages = {183-191}, pmid = {29491905}, issn = {1674-5507}, abstract = {The reinforcement of premating barriers due to reduced hybrid fitness in sympatry may cause secondary sexual isolation within a species as a by-product. Consistent with this, in the fly Drosophila subquinaria, females that are sympatric with D. recens mate at very low rates not only with D. recens, but also with conspecific D. subquinaria males from allopatry. Here, we ask if these effects of reinforcement cascade more broadly to affect sexual isolation with other closely related species. We assay reproductive isolation of these species with D. transversa and find that choosy D. subquinaria females from the region sympatric with D. recens discriminate strongly against male D. transversa, whereas D. subquinaria from the allopatric region do not. This increased sexual isolation cannot be explained by natural selection to avoid mating with this species, as they are allopatric in geographic range and we do not identify any intrinsic postzygotic isolation between D. subquinaria and D. transversa. Variation in epicuticular hydrocarbons, which are used as mating signals in D. subquinaria, follow patterns of premating isolation: D. transversa and allopatric D. subquinaria are most similar to each other and differ from sympatric D. subquinaria, and those of D. recens are distinct from the other two species. We suggest that the secondary effects of reinforcement may cascade to strengthen reproductive isolation with other species that were not a target of selection. These effects may enhance the divergence that occurs in allopatry to help explain why some species are already sexually isolated upon secondary contact.}, } @article {pmid29588003, year = {2015}, author = {Lees, RS and Gilles, JR and Hendrichs, J and Vreysen, MJ and Bourtzis, K}, title = {Back to the future: the sterile insect technique against mosquito disease vectors.}, journal = {Current opinion in insect science}, volume = {10}, number = {}, pages = {156-162}, doi = {10.1016/j.cois.2015.05.011}, pmid = {29588003}, issn = {2214-5753}, abstract = {With the global burden of mosquito-borne diseases increasing, and some conventional vector control tools losing effectiveness, the sterile insect technique (SIT) is a potential new tool in the arsenal. Equipment and protocols have been developed and validated for efficient mass-rearing, irradiation and release of Aedines and Anophelines that could be useful for several control approaches. Assessment of male quality is becoming more sophisticated, and several groups are well advanced in pilot site selection and population surveillance. It will not be long before SIT feasibility has been evaluated in various settings. Until perfect sexing mechanisms exist, combination of Wolbachia-induced phenotypes, such as cytoplasmic incompatibility and pathogen interference, and irradiation may prove to be the safest solution for population suppression.}, } @article {pmid27517272, year = {2008}, author = {Hoerauf, A and Specht, S and Büttner, M and Pfarr, K and Mand, S and Fimmers, R and Marfo-Debrekyei, Y and Konadu, P and Debrah, AY and Bandi, C and Brattig, N and Albers, A and Larbi, J and Batsa, L and Taylor, MJ and Adjei, O and Büttner, DW}, title = {Wolbachia endobacteria depletion by doxycycline as antifilarial therapy has macrofilaricidal activity in onchocerciasis: a randomized placebo-controlled study.}, journal = {Medical microbiology and immunology}, volume = {197}, number = {3}, pages = {335}, doi = {10.1007/s00430-007-0072-z}, pmid = {27517272}, issn = {0300-8584}, } @article {pmid28565520, year = {1999}, author = {Shoemaker, DD and Katju, V and Jaenike, J}, title = {WOLBACHIA AND THE EVOLUTION OF REPRODUCTIVE ISOLATION BETWEEN DROSOPHILA RECENS AND DROSOPHILA SUBQUINARIA.}, journal = {Evolution; international journal of organic evolution}, volume = {53}, number = {4}, pages = {1157-1164}, doi = {10.1111/j.1558-5646.1999.tb04529.x}, pmid = {28565520}, issn = {1558-5646}, abstract = {Endosymbiotic bacteria of the genus Wolbachia are widespread among insects and in many cases cause cytoplasmic incompatibility in crosses between infected males and uninfected females. Such findings have been used to argue that Wolbachia have played an important role in insect speciation. Theoretical models, however, indicate that Wolbachia alone are unlikely to lead to stable reproductive isolation between two formerly conspecific populations. Here we analyze the components of reproductive isolation between Drosophila recens, which is infected with Wolbachia, and its uninfected sister species Drosophila subquinaria. Laboratory pairings demonstrated that gene flow via matings between D. recens females and D. subquinaria males is hindered by behavioral isolation. Matings readily occurred in the reciprocal cross (D. quinaria females × D. recens males), but very few viable progeny were produced. The production of viable hybrids via this route was restored by antibiotic curing of D. recens of their Wolbachia symbionts, indicating that hybrid offspring production is greatly reduced by cytoplasmic incompatibility in the crosses involving infected D. recens males. Thus, behavioral isolation and Wolbachia-induced cytoplasmic incompatibility act as complementary asymmetrical isolating mechanisms between these two species. In accordance with Haldane's rule, hybrid females were fertile, whereas hybrid males invariably were sterile. Levels of mtDNA variation in D. recens are much lower than in either D. subquinaria or D. falleni, neither of which is infected with Wolbachia. The low haplotype diversity in D. recens is likely due to an mtDNA sweep associated with the spread of Wolbachia. Nevertheless, the existence of several mtDNA haplotypes in this species indicates that Wolbachia have been present as a potential isolating mechanism for a substantial period of evolutionary time. Finally, we argue that although Wolbachia by themselves are unlikely to bring about speciation, they can increase the rate of speciation in insects.}, } @article {pmid28565514, year = {1999}, author = {Belshaw, R and Quicke, DLJ and Völkl, W and Godfray, HCJ}, title = {MOLECULAR MARKERS INDICATE RARE SEX IN A PREDOMINANTLY ASEXUAL PARASITOID WASP.}, journal = {Evolution; international journal of organic evolution}, volume = {53}, number = {4}, pages = {1189-1199}, doi = {10.1111/j.1558-5646.1999.tb04532.x}, pmid = {28565514}, issn = {1558-5646}, abstract = {The parasitoid wasp genus Lysiphlebus (Hymenoptera: Braconidae: Aphidiinae) contains a taxonomically poorly resolved group of both sexual (arrhenotokous) species and asexual (thelytokous) clones. Maximum-parsimony and maximum-likelihood analyses of mitochondrial DNA sequence data from specimens collected across Western Europe showed that asexuality, which does not appear to be caused by the bacterium Wolbachia, is concentrated in two geographically widespread lineages, the older of which diverged from the closest extant sexual taxa approximately 0.5 million years ago. However, the DNA sequences of a nuclear intron (elongation factor-1α) showed no congruence with this pattern, and a much higher frequency of heterozygotes with very high allelic diversity was observed among the asexual females compared to that among females from the sexual species. This pattern is consistent with maternally inherited asexuality coupled with a history of rare sex with members of several closely related sexual populations or species. Our observations reinforce recent arguments that rare sex may be more important for the persistence of otherwise asexual lineages than hitherto appreciated.}, } @article {pmid28568785, year = {1997}, author = {Poinsot, D and Merçot, H}, title = {WOLBACHIA INFECTION IN DROSOPHILA SIMULANS: DOES THE FEMALE HOST BEAR A PHYSIOLOGICAL COST?.}, journal = {Evolution; international journal of organic evolution}, volume = {51}, number = {1}, pages = {180-186}, doi = {10.1111/j.1558-5646.1997.tb02399.x}, pmid = {28568785}, issn = {1558-5646}, abstract = {Fitness traits of three Drosophila simulans strains infected by endocellular bacteria belonging to the genus Wolbachia have been compared with those of replicate stocks previously cured from the infection by an antibiotic treatment. The traits measured were development time, egg-to-adult viability, egg hatch, productivity, fecundity, and the number of functional ovarioles. Individuals of the first strain were bi-infected by two Wolbachia variants, wHa and wNo. The second strain was infected by wHa, the third one by wNo. The Wolbachia studied here cause cytoplasmic incompatibility (CI), a high embryonic mortality (70% to > 90%) when an infected male is crossed with an uninfected female. Three generations after antibiotic treatment, we observed in all strains a significant drop in productivity in the cured stocks. This drop was not due to antibiotic toxicity and was associated with the loss of the Wolbachia. However the effect had disappeared in two of the three strains five generations after treatment, and could not be found in the third strain in a third measurement carried out 14 generations after treatment. The temporary nature of the productivity difference indicates that Wolbachia do not enhance productivity in infected strains. On the other hand, in all traits measured, our results show the absence of any negative effects of the Wolbachia on their host. This could be explained when considering Wolbachia evolution, as maternally transmitted parasites bear a strong selective pressure not to harm their female host. However, CI would allow the bacteria to be maintained even when harming the female. The apparent absence of deleterious effects caused by these Wolbachia might result from a trade-off, where a relatively low bacteria density would advantage the Wolbachia by suppressing any deleterious effects on the female host, at the cost of a weaker maternal transmission rate of the infection.}, } @article {pmid28568927, year = {1996}, author = {Orr, MR}, title = {LIFE-HISTORY ADAPTATION AND REPRODUCTIVE ISOLATION IN A GRASSHOPPER HYBRID ZONE.}, journal = {Evolution; international journal of organic evolution}, volume = {50}, number = {2}, pages = {704-716}, doi = {10.1111/j.1558-5646.1996.tb03880.x}, pmid = {28568927}, issn = {1558-5646}, abstract = {Patterns of life-history adaptation and reproductive isolation were investigated in the acridid grasshoppers Melanoplus sanguinipes and M. devastator, which hybridize along an altitudinal gradient in the Sierra Nevada of California. Melanoplus sanguinipes females crossed with M. devastator males produced eggs that were approximately half as viable as eggs from other crosses. Diminished viability was not attributable either to infection by Wolbachia pipientis or to failure of sperm transfer. When offered an opportunity to choose a mate, females from all populations discriminated against males of the other species, whereas in no-choice tests measuring copulation duration only females from the tails of the clines showed preferences. Melanoplus sanguinipes, found at high elevations where the growing season is short, exhibited faster egg hatch, faster larval development, smaller adult body sizes, and smaller clutch sizes than M. devastator. Melanoplus devastator, from California's Central Valley, endured a hot and dry summer in a reproductive diapause that was absent in M. sanguinipes. Clines in reproductive diapause and clutch size coincided with the region of reproductive incompatibility. Development time, body size, and hatch time also changed across the hybrid zone, but the regions of largest transitions in these traits were either difficult to locate using the limited populations studied here or were not coincident with the zone's center. A method is described for combining ecological and phylogenetic analyses to address the unknown issue of whether life-history divergence has conributed to reproductive isolation in this system.}, } @article {pmid28568404, year = {1994}, author = {Turelli, M}, title = {EVOLUTION OF INCOMPATIBILITY-INDUCING MICROBES AND THEIR HOSTS.}, journal = {Evolution; international journal of organic evolution}, volume = {48}, number = {5}, pages = {1500-1513}, doi = {10.1111/j.1558-5646.1994.tb02192.x}, pmid = {28568404}, issn = {1558-5646}, abstract = {In many insect species, males infected with microbes related to Wolbachia pipientis are "incompatible" with uninfected females. Crosses between infected males and uninfected females produce significantly fewer adult progeny than the other three possible crosses. The incompatibility-inducing microbes are usually maternally transmitted. Thus, incompatibility tends to confer a reproductive advantage on infected females in polymorphic populations, allowing these infections to spread. This paper analyzes selection on parasite and host genes that affect such incompatibility systems. Selection among parasite variants does not act directly on the level of incompatibility with uninfected females. In fact, selection favors rare parasite variants that increase the production of infected progeny by infected mothers, even if these variants reduce incompatibility with uninfected females. However, productivity-reducing parasites that cause partial incompatibility with hosts harboring alternative variants can be favored once they become sufficiently abundant locally. Thus, they may spread spatially by a process analogous to the spread of underdominant chromosome rearrangements. The dynamics of modifier alleles in the host are more difficult to predict, because such alleles will occur in both infected and uninfected individuals. Nevertheless, the relative fecundity of infected females compared to uninfected females, the efficiency of maternal transmission and the mutual compatibility of infected individuals all tend to increase under within-population selection on both host and parasite genes. In addition, selection on host genes favors increased compatibility between infected males and uninfected females. Although vertical transmission tends to harmonize host and parasite evolution, competition among parasite variants will tend to maintain incompatibility.}, } @article {pmid28568258, year = {1994}, author = {Prout, T}, title = {SOME EVOLUTIONARY POSSIBILITIES FOR A MICROBE THAT CAUSES INCOMPATIBILITY IN ITS HOST.}, journal = {Evolution; international journal of organic evolution}, volume = {48}, number = {3}, pages = {909-911}, doi = {10.1111/j.1558-5646.1994.tb01371.x}, pmid = {28568258}, issn = {1558-5646}, } @article {pmid28568710, year = {1993}, author = {Kambhampati, S and Rai, KS and Burgun, SJ}, title = {UNIDIRECTIONAL CYTOPLASMIC INCOMPATIBILITY IN THE MOSQUITO, AEDES ALBOPICTUS.}, journal = {Evolution; international journal of organic evolution}, volume = {47}, number = {2}, pages = {673-677}, doi = {10.1111/j.1558-5646.1993.tb02121.x}, pmid = {28568710}, issn = {1558-5646}, } @article {pmid27126028, year = {2016}, author = {O'Neill, SL}, title = {Wolbachia mosquito control: Tested.}, journal = {Science (New York, N.Y.)}, volume = {352}, number = {6285}, pages = {526}, doi = {10.1126/science.352.6285.526-a}, pmid = {27126028}, issn = {1095-9203}, mesh = {Animals ; Culicidae/*microbiology ; Dengue/*prevention & control ; Mosquito Control/*methods ; *Wolbachia ; }, } @article {pmid27124663, year = {2016}, author = {Aliota, MT and Walker, EC and Uribe Yepes, A and Velez, ID and Christensen, BM and Osorio, JE}, title = {The wMel Strain of Wolbachia Reduces Transmission of Chikungunya Virus in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {4}, pages = {e0004677}, pmid = {27124663}, issn = {1935-2735}, support = {R21 AI117413/AI/NIAID NIH HHS/United States ; #AI117413-01/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; *Antibiosis ; Chikungunya Fever/prevention & control/*transmission ; Chikungunya virus/*isolation & purification ; Disease Transmission, Infectious/*prevention & control ; Insect Vectors ; Saliva/virology ; Survival Analysis ; Wolbachia/*growth & development/physiology ; }, abstract = {BACKGROUND: New approaches to preventing chikungunya virus (CHIKV) are needed because current methods are limited to controlling mosquito populations, and they have not prevented the invasion of this virus into new locales, nor have they been sufficient to control the virus upon arrival. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against CHIKV. Although this approach holds much promise for limiting virus transmission, at present our understanding of the ability of CHIKV to infect, disseminate, and be transmitted by wMel-infected Ae. aegypti currently being used at Wolbachia release sites is limited.

Using Ae. aegypti infected with the wMel strain of Wolbachia that are being released in Medellin, Colombia, we report that these mosquitoes have reduced vector competence for CHIKV, even with extremely high viral titers in the bloodmeal. In addition, we examined the dynamics of CHIKV infection over the course of four to seven days post feeding. Wolbachia-infected mosquitoes remained non-infective over the duration of seven days, i.e., no infectious virus was detected in the saliva when exposed to bloodmeals of moderate viremia, but CHIKV-exposed, wild type mosquitoes did have viral loads in the saliva consistent with what has been reported elsewhere. Finally, the presence of wMel infection had no impact on the lifespan of mosquitoes as compared to wild type mosquitoes following CHIKV infection.

CONCLUSIONS/SIGNIFICANCE: These results could have an impact on vector control strategies in areas where Ae. aegypti are transmitting both DENV and CHIKV; i.e., they argue for further exploration, both in the laboratory and the field, on the feasibility of expanding this technology beyond DENV.}, } @article {pmid27122079, year = {2016}, author = {Wallau, GL and da Rosa, MT and De Ré, FC and Loreto, EL}, title = {Wolbachia from Drosophila incompta: just a hitchhiker shared by Drosophila in the New and Old World?.}, journal = {Insect molecular biology}, volume = {25}, number = {4}, pages = {487-499}, doi = {10.1111/imb.12237}, pmid = {27122079}, issn = {1365-2583}, mesh = {Animals ; Brazil ; Drosophila/*microbiology ; *Evolution, Molecular ; *Genome, Bacterial ; Phylogeny ; Wolbachia/*genetics/physiology ; }, abstract = {Wolbachia are intracellular endosymbionts that infect arthropods and filarial nematodes, occasionally causing a wide variety of modifications in host biology, such as male-killing and cytoplasmic incompatibility (CI), amongst others. This study assembled draft genomes for Wolbachia infecting Drosophila incompta, a species that uses flowers as exclusive breeding and feeding sites, in two distinct Brazilian populations. The absence of four genes involved in CI from this genome, together with literature reports of low frequencies of infected flies in wild populations that contain high mitogenome polymorphism, suggests that this bacterium does not induce CI in D. incompta. Phylogenomic analysis placed Wolbachia infecting D. incompta as closely related to the wMel strain which received such name since it was originally detected in Drosophila melanogaster. In addition, phylogenetic analysis using the Wolbachia surface protein gene and five genes used for multilocus sequence typing of Wolbachia found infecting Drosophila and other arthropod species of Old and New World displayed a complex evolutionary scenario involving recent horizontal transfer bursts in all major clades of Wolbachia pipens belonging to the supergroup A in both geographical regions.}, } @article {pmid27114607, year = {2016}, author = {Li, YY and Fields, PG and Pang, BP and Floate, KD}, title = {Effects of Tetracycline and Rifampicin Treatments on the Fecundity of the Wolbachia -Infected Host, Tribolium confusum (Coleoptera: Tenebrionidae).}, journal = {Journal of economic entomology}, volume = {109}, number = {3}, pages = {1458-1464}, doi = {10.1093/jee/tow067}, pmid = {27114607}, issn = {1938-291X}, abstract = {We examined the effects of Wolbachia bacteria on the reproduction of the flour beetle Tribolium confusum (Coleoptera: Tenebrionidae) using different antibiotics and across generations. We first removed infections by rearing insects on a diet with tetracycline (T; 1.0, 2.0, 3.0, 5.0, 10.0 mg/g) or rifampicin (R; 0.1, 0.2, 0.3, 0.5, 1.0 mg/g). We then performed experimental crosses using adults two generations (G2) and four generations (G4) removed from antibiotic treatments. Results showed that use of rifampicin more readily cured infections. Egg hatch from crosses of uninfected females and infected males was 0, but averaged 84 to 91% for eggs from all other crosses. Elevated fecundity was observed for T-G2 females, but not for T-G4, R-G2, or R-G4 females. Cross type had little or no effect on the sex of F 1 offspring, which averaged 52% female. These collective results support previous findings that show that Wolbachia in T. confusum causes 100% cytoplasmic incompatibility and emphasize that the antibiotic treatment used to remove infections may have additional consequences (e.g., elevated fecundity) that may not be apparent in subsequent generations.}, } @article {pmid27110057, year = {2016}, author = {Libro, S and Slatko, BE and Foster, JM}, title = {Characterization of innate immunity genes in the parasitic nematode Brugia malayi.}, journal = {Symbiosis (Philadelphia, Pa.)}, volume = {68}, number = {}, pages = {145-155}, pmid = {27110057}, issn = {0334-5114}, abstract = {The filarial nematode Brugia malayi is one of the causative agents of lymphatic filariasis, a neglected tropical disease that affects 120 million people worldwide. The limited effectiveness of available anthelmintics and the absence of a vaccine have prompted extensive research on the interaction between Brugia and its obligate bacterial endosymbiont, Wolbachia. Recent studies suggest that Wolbachia is able to manipulate its nematode host immunity but relatively little is known about the immune system of filarial nematodes. Therefore, elucidation of the mechanisms underlying the immune system of B. malayi may be useful for understanding how the symbiotic relationship is maintained and help in the identification of new drug targets. In order to characterize the main genetic pathways involved in B. malayi immunity, we exposed adult female worms to two bacterial lysates (Escherichia coli and Bacillus amyloliquefaciens), dsRNA and dsDNA. We performed transcriptome sequencing of worms exposed to each immune elicitor at two different timepoints. Gene expression analysis of untreated and immune-challenged worms was performed to characterize gene expression patterns associated with each type of immune stimulation. Our results indicate that different immune elicitors produced distinct expression patterns in B. malayi, with changes in the expression of orthologs of well-characterized C. elegans immune pathways such as insulin, TGF-β, and p38 MAPK pathways, as well as C-type lectins and several stress-response genes.}, } @article {pmid27106932, year = {2016}, author = {Kho, EA and Hugo, LE and Lu, G and Smith, DD and Kay, BH}, title = {Effects of Larval Nutrition on Wolbachia-Based Dengue Virus Interference in Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {53}, number = {4}, pages = {894-901}, doi = {10.1093/jme/tjw029}, pmid = {27106932}, issn = {1938-2928}, mesh = {Aedes/growth & development/*physiology/*virology ; *Animal Nutritional Physiological Phenomena ; Animals ; Dengue Virus/physiology ; Diet ; Female ; Insect Vectors/growth & development/*physiology/*virology ; Larva/growth & development/physiology/virology ; Mosquito Control ; Pest Control, Biological ; Wolbachia/*physiology ; }, abstract = {In order to assess the broad-scale applicability of field releases of Wolbachia for the biological control of insect-transmitted diseases, we determined the relationship between the larval diet of Aedes aegypti L. mosquitoes infected with Wolbachia strains and their susceptibility to dengue virus (DENV) infection via intrathoracic injection and oral inoculation. Larvae were reared on diets that varied in the quantity of food which had the effect of modifying development time and adult body size. Wolbachia wMel infection was associated with highly significant reductions in dengue serotype 2 (DENV-2) infection rates of between 80 and 97.5% following intrathoracic injection of adults emerging from three diet levels. Reductions were 100% in two diet level treatments following oral inoculation. Similarly, wMelPop infection was associated with highly significant reductions in DENV-2 infection rates of between 95 and 100% for intrathoracic injection and 97.5 and 100% for oral inoculation across diet level treatments. Larval diet level had no significant effect on DENV-2 infection rates in the presence of Wolbachia infection in mosquitoes that were intrathoracically injected with the virus. This indicates that the effectiveness of Wolbachia on vector competence disruption within Ae. aegypti is unlikely to be compromised by variable larval nutrition in field settings.}, } @article {pmid27097518, year = {2016}, author = {Ndii, MZ and Allingham, D and Hickson, RI and Glass, K}, title = {The effect of Wolbachia on dengue dynamics in the presence of two serotypes of dengue: symmetric and asymmetric epidemiological characteristics.}, journal = {Epidemiology and infection}, volume = {144}, number = {13}, pages = {2874-2882}, pmid = {27097518}, issn = {1469-4409}, mesh = {Aedes/*microbiology/virology ; Animals ; Coinfection/prevention & control/transmission/virology ; Dengue/*epidemiology/prevention & control/transmission/virology ; Dengue Virus/genetics/*physiology ; Humans ; Incidence ; Insect Vectors/*microbiology/virology ; Models, Theoretical ; Serogroup ; Wolbachia ; }, abstract = {An innovative strategy to reduce dengue transmission uses the bacterium Wolbachia. We analysed the effects of Wolbachia on dengue transmission dynamics in the presence of two serotypes of dengue using a mathematical model, allowing for differences in the epidemiological characteristics of the serotypes. We found that Wolbachia has a greater effect on secondary infections than on primary infections across a range of epidemiological characteristics. If one serotype is more transmissible than the other, it will dominate primary infections and Wolbachia will be less effective at reducing secondary infections of either serotype. Differences in the antibody-dependent enhancement of the two serotypes have considerably less effect on the benefits of Wolbachia than differences in transmission probability. Even if the antibody-dependent enhancement rate is high, Wolbachia is still effective in reducing dengue. Our findings suggest that Wolbachia will be effective in the presence of more than one serotype of dengue; however, a better understanding of serotype-specific differences in transmission probability may be needed to optimize delivery of a Wolbachia intervention.}, } @article {pmid27095842, year = {2016}, author = {Hopkin, K}, title = {Inner Workings: A big role for a microbial parasite.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {113}, number = {16}, pages = {4236-4237}, pmid = {27095842}, issn = {1091-6490}, mesh = {Animals ; Drosophila melanogaster ; Host-Pathogen Interactions/*physiology ; Wolbachia/*physiology ; }, } @article {pmid27090067, year = {2017}, author = {Kishani Farahani, H and Ashouri, A and Goldansaz, SH and Shapiro, MS and Pierre, JS and van Baaren, J}, title = {Decrease of memory retention in a parasitic wasp: an effect of host manipulation by Wolbachia?.}, journal = {Insect science}, volume = {24}, number = {4}, pages = {569-583}, doi = {10.1111/1744-7917.12348}, pmid = {27090067}, issn = {1744-7917}, mesh = {Animals ; *Association Learning ; *Behavior, Animal ; Female ; *Memory ; Moths/parasitology ; Odorants ; Oviposition/physiology ; Wasps/*microbiology/*physiology ; *Wolbachia ; }, abstract = {Several factors, such as cold exposure, aging, the number of experiences and viral infection, have been shown to affect learning ability in different organisms. Wolbachia has been found worldwide as an arthropod parasite/mutualist symbiont in a wide range of species, including insects. Differing effects have been identified on physiology and behavior by Wolbachia. However, the effect of Wolbachia infection on the learning ability of their host had never previously been studied. The current study carried out to compare learning ability and memory duration in 2 strains of the parasitoid Trichogramma brassicae: 1 uninfected and 1 infected by Wolbachia. Both strains were able to associate the novel odors with the reward of an oviposition into a host egg. However, the percentage of females that responded to the experimental design and displayed an ability to learn in these conditions was higher in the uninfected strain. Memory duration was longer in uninfected wasps (23.8 and 21.4 h after conditioning with peppermint and lemon, respectively) than in infected wasps (18.9 and 16.2 h after conditioning with peppermint and lemon, respectively). Memory retention increased in response to the number of conditioning sessions in both strains, but memory retention was always shorter in the infected wasps than in the uninfected ones. Wolbachia infection may select for reduced memory retention because shorter memory induces infected wasps to disperse in new environments and avoid competition with uninfected wasps by forgetting cues related to previously visited environments, thus increasing transmission of Wolbachia in new environments.}, } @article {pmid27089431, year = {2016}, author = {Rainey, SM and Martinez, J and McFarlane, M and Juneja, P and Sarkies, P and Lulla, A and Schnettler, E and Varjak, M and Merits, A and Miska, EA and Jiggins, FM and Kohl, A}, title = {Wolbachia Blocks Viral Genome Replication Early in Infection without a Transcriptional Response by the Endosymbiont or Host Small RNA Pathways.}, journal = {PLoS pathogens}, volume = {12}, number = {4}, pages = {e1005536}, pmid = {27089431}, issn = {1553-7374}, support = {094664MA/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/MRC_/Medical Research Council/United Kingdom ; MC_UP_1102/13/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; MC_UP_A550_1031/MRC_/Medical Research Council/United Kingdom ; /CRUK_/Cancer Research UK/United Kingdom ; }, mesh = {Animals ; Cell Line ; Drosophila melanogaster/*genetics ; *Genome, Viral/genetics ; Humans ; MicroRNAs/genetics ; RNA, Small Interfering/genetics ; RNA, Viral/*genetics ; Semliki forest virus ; Symbiosis ; Transcription, Genetic ; Virus Replication/*physiology ; Wolbachia/*metabolism ; }, abstract = {The intracellular endosymbiotic bacterium Wolbachia can protect insects against viral infection, and is being introduced into mosquito populations in the wild to block the transmission of arboviruses that infect humans and are a major public health concern. To investigate the mechanisms underlying this antiviral protection, we have developed a new model system combining Wolbachia-infected Drosophila melanogaster cell culture with the model mosquito-borne Semliki Forest virus (SFV; Togaviridae, Alphavirus). Wolbachia provides strong antiviral protection rapidly after infection, suggesting that an early stage post-infection is being blocked. Wolbachia does appear to have major effects on events distinct from entry, assembly or exit as it inhibits the replication of an SFV replicon transfected into the cells. Furthermore, it causes a far greater reduction in the expression of proteins from the 3' open reading frame than the 5' non-structural protein open reading frame, indicating that it is blocking the replication of viral RNA. Further to this separation of the replicase proteins and viral RNA in transreplication assays shows that uncoupling of viral RNA and replicase proteins does not overcome Wolbachia's antiviral activity. This further suggests that replicative processes are disrupted, such as translation or replication, by Wolbachia infection. This may occur by Wolbachia mounting an active antiviral response, but the virus did not cause any transcriptional response by the bacterium, suggesting that this is not the case. Host microRNAs (miRNAs) have been implicated in protection, but again we found that host cell miRNA expression was unaffected by the bacterium and neither do our findings suggest any involvement of the antiviral siRNA pathway. We conclude that Wolbachia may directly interfere with early events in virus replication such as translation of incoming viral RNA or RNA transcription, and this likely involves an intrinsic (as opposed to an induced) mechanism.}, } @article {pmid27078260, year = {2016}, author = {Voronin, D and Bachu, S and Shlossman, M and Unnasch, TR and Ghedin, E and Lustigman, S}, title = {Glucose and Glycogen Metabolism in Brugia malayi Is Associated with Wolbachia Symbiont Fitness.}, journal = {PloS one}, volume = {11}, number = {4}, pages = {e0153812}, pmid = {27078260}, issn = {1932-6203}, support = {R01 AI072465/AI/NIAID NIH HHS/United States ; R56 AI101372/AI/NIAID NIH HHS/United States ; AI072465/AI/NIAID NIH HHS/United States ; R56AI101372/AI/NIAID NIH HHS/United States ; }, mesh = {Aldehyde-Lyases/genetics/metabolism ; Animals ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/metabolism ; Blotting, Western ; Brugia malayi/genetics/*metabolism/*microbiology ; Doxycycline/pharmacology ; Female ; Gene Expression ; Glucose/*metabolism ; Glycogen/*metabolism ; Glycolysis ; Helminth Proteins/genetics/metabolism ; Host-Pathogen Interactions/drug effects ; Microscopy, Confocal ; Microscopy, Electron, Transmission ; Microscopy, Immunoelectron ; RNA Interference ; Reverse Transcriptase Polymerase Chain Reaction ; Symbiosis/drug effects ; Wolbachia/metabolism/*physiology/ultrastructure ; }, abstract = {Wolbachia are endosymbiotic bacteria found in the majority of arthropods and filarial nematodes of medical and veterinary importance. They have evolved a wide range of symbiotic associations. In filarial nematodes that cause human lymphatic filariasis (Wuchereria bancrofti, Brugia malayi) or onchocerciasis (Onchocerca volvulus), Wolbachia are important for parasite development, reproduction and survival. The symbiotic bacteria rely in part on nutrients and energy sources provided by the host. Genomic analyses suggest that the strain of Wolbachia found in B. malayi (wBm) lacks the genes for two glycolytic enzymes--6-phosphofructokinase and pyruvate kinase--and is thus potentially unable to convert glucose into pyruvate, an important substrate for energy generation. The Wolbachia surface protein, wBm00432, is complexed to six B. malayi glycolytic enzymes, including aldolase. In this study we characterized two B. malayi aldolase isozymes and found that their expression is dependent on Wolbachia fitness and number. We confirmed by immuno-transmission electron microscopy that aldolase is associated with the Wolbachia surface. RNAi experiments suggested that aldolase-2 plays a significant role in both Wolbachia survival and embryogenesis in B. malayi. Treatment with doxycycline reduced Wolbachia fitness and increased the amount of both glucose and glycogen detected in the filarial parasite, indicating that glucose metabolism and glycogen storage in B. malayi are associated with Wolbachia fitness. This metabolic co-dependency between Wolbachia and its filarial nematode indicates that glycolysis could be a shared metabolic pathway between the bacteria and B. malayi, and thus a potential new target for anti-filarial therapy.}, } @article {pmid27076356, year = {2016}, author = {Kriesner, P and Conner, WR and Weeks, AR and Turelli, M and Hoffmann, AA}, title = {Persistence of a Wolbachia infection frequency cline in Drosophila melanogaster and the possible role of reproductive dormancy.}, journal = {Evolution; international journal of organic evolution}, volume = {70}, number = {5}, pages = {979-997}, pmid = {27076356}, issn = {1558-5646}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Africa ; Animals ; Australia ; Drosophila melanogaster/genetics/*microbiology/physiology ; Ecosystem ; Female ; Fertility ; Genetic Fitness ; *Host-Pathogen Interactions ; Male ; North America ; Tropical Climate ; Wolbachia/*pathogenicity ; }, abstract = {Field populations of arthropods are often polymorphic for Wolbachia but the factors maintaining intermediate Wolbachia frequencies are generally not understood. In Drosophila melanogaster, Wolbachia frequencies are highly variable across the globe. We document the persistence of a Wolbachia infection frequency cline in D. melanogaster populations from eastern Australia across at least 20 years, with frequencies generally high in the tropics but lower in cool temperate regions. The results are interpreted using a model of frequency dynamics incorporating cytoplasmic incompatibility (CI), imperfect maternal transmission and Wolbachia effects on fitness. Clinal variation is less pronounced in eastern North America which may reflect annual recolonization at higher latitudes. Limited samples from Africa from latitudes matching our tropical and subtropical samples from Australia and North America show comparably high infection frequencies, but some equatorial samples show lower frequencies. Adult dormancy across cold periods may contribute to the Australian Wolbachia cline. Infected flies exposed to cold conditions for an extended period had reduced fecundity and viability, an effect not evident in unexposed controls. These fitness costs may contribute to the relatively low Wolbachia frequencies in Australian temperate areas; whereas different processes, including CI induced by young males, may contribute to higher frequencies in tropical locations.}, } @article {pmid27069790, year = {2016}, author = {Lefoulon, E and Bain, O and Makepeace, BL and d'Haese, C and Uni, S and Martin, C and Gavotte, L}, title = {Breakdown of coevolution between symbiotic bacteria Wolbachia and their filarial hosts.}, journal = {PeerJ}, volume = {4}, number = {}, pages = {e1840}, pmid = {27069790}, issn = {2167-8359}, abstract = {Wolbachia is an alpha-proteobacterial symbiont widely distributed in arthropods. Since the identification of Wolbachia in certain animal-parasitic nematodes (the Onchocercidae or filariae), the relationship between arthropod and nematode Wolbachia has attracted great interest. The obligate symbiosis in filariae, which renders infected species susceptible to antibiotic chemotherapy, was held to be distinct from the Wolbachia-arthropod relationship, typified by reproductive parasitism. While co-evolutionary signatures in Wolbachia-arthropod symbioses are generally weak, reflecting horizontal transmission events, strict co-evolution between filariae and Wolbachia has been reported previously. However, the absence of close outgroups for phylogenetic studies prevented the determination of which host group originally acquired Wolbachia. Here, we present the largest co-phylogenetic analysis of Wolbachia in filariae performed to date including: (i) a screening and an updated phylogeny of Wolbachia; (ii) a co-phylogenetic analysis; and (iii) a hypothesis on the acquisition of Wolbachia infection. First, our results show a general overestimation of Wolbachia occurrence and support the hypothesis of an ancestral absence of infection in the nematode phylum. The accuracy of supergroup J is also underlined. Second, although a global pattern of coevolution remains, the signal is derived predominantly from filarial clades associated with Wolbachia in supergroups C and J. In other filarial clades, harbouring Wolbachia supergroups D and F, horizontal acquisitions and secondary losses are common. Finally, our results suggest that supergroup C is the basal Wolbachia clade within the Ecdysozoa. This hypothesis on the origin of Wolbachia would change drastically our understanding of Wolbachia evolution.}, } @article {pmid27065360, year = {2016}, author = {Sahoo, RK}, title = {Why Antagonistic Traits against Cytoplasmic Incompatibility Are So Elusive.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {392}, pmid = {27065360}, issn = {1664-302X}, } @article {pmid27055293, year = {2015}, author = {Goryacheva, II and Gorelova, TV and Andrianov, BV}, title = {[Drosophila melanogaster Cell Culture as an Experimental Model to Study Recombination in Wolbachia pipientis].}, journal = {Genetika}, volume = {51}, number = {12}, pages = {1345-1350}, pmid = {27055293}, issn = {0016-6758}, mesh = {Animals ; Cell Culture Techniques/*methods ; Cells, Cultured ; Drosophila melanogaster ; *Recombination, Genetic ; Wolbachia/*genetics/*growth & development ; }, abstract = {Wolbachiapipientis is an obligate intracellular endosymbiont that commonly infects arthropods. Comparative genomic studies of Wolbachia reveal traces of numerous events of intergenic and intragenic recombination. The molecular mechanisms of recombination in Wolbachia are not currently known. We conducted experimental verification of the possibility of recombination of two strains of Wolbachia: wMel and wRi, after using these strains for double infection of the Dm2008Wb1 (D. melanogaster) cell culture clone permissive to Wolbachia. We obtained cell culture subclones with double Wolbachia infection and subclones infected only by strain wMel. Dual infection with the Wolbachia strains wMel and wRi has been stably maintained in the subclones for two years. Multilocus sequence typing (MLST) of the obtained subclones revealed the presence of dual infection for all five Wolbachia genes used for MLST Cloning and nucleotide sequence analysis of individual forms of the fbpA gene of Wolbachia from cell clones with dual infection showed intragenic recombination events between strains wMel and wRi, which occurred in the permanent D. melanogaster culture cell culture. The fact that putative recombination sites contain no insertions of nucleotide sequences of phages or IS elements, as well as the asymmetrical character of recombinants, favors the hypothesis that gene conversion is the most probable molecular mechanism of recombination in Wolbachia.}, } @article {pmid27047911, year = {2016}, author = {Murray, JV and Jansen, CC and De Barro, P}, title = {Risk Associated with the Release of Wolbachia-Infected Aedes aegypti Mosquitoes into the Environment in an Effort to Control Dengue.}, journal = {Frontiers in public health}, volume = {4}, number = {}, pages = {43}, pmid = {27047911}, issn = {2296-2565}, abstract = {BACKGROUND: In an effort to eliminate dengue, a successful technology was developed with the stable introduction of the obligate intracellular bacteria Wolbachia pipientis into the mosquito Aedes aegypti to reduce its ability to transmit dengue fever due to life shortening and inhibition of viral replication effects. An analysis of risk was required before considering release of the modified mosquito into the environment.

METHODS: Expert knowledge and a risk assessment framework were used to identify risk associated with the release of the modified mosquito. Individual and group expert elicitation was performed to identify potential hazards. A Bayesian network (BN) was developed to capture the relationship between hazards and the likelihood of events occurring. Risk was calculated from the expert likelihood estimates populating the BN and the consequence estimates elicited from experts.

RESULTS: The risk model for "Don't Achieve Release" provided an estimated 46% likelihood that the release would not occur by a nominated time but generated an overall risk rating of very low. The ability to obtain compliance had the greatest influence on the likelihood of release occurring. The risk model for "Cause More Harm" provided a 12.5% likelihood that more harm would result from the release, but the overall risk was considered negligible. The efficacy of mosquito management had the most influence, with the perception that the threat of dengue fever had been eliminated, resulting in less household mosquito control, and was scored as the highest ranked individual hazard (albeit low risk).

CONCLUSIONS: The risk analysis was designed to incorporate the interacting complexity of hazards that may affect the release of the technology into the environment. The risk analysis was a small, but important, implementation phase in the success of this innovative research introducing a new technology to combat dengue transmission in the environment.}, } @article {pmid27035832, year = {2016}, author = {Hunter, CM and Huang, W and Mackay, TF and Singh, ND}, title = {The Genetic Architecture of Natural Variation in Recombination Rate in Drosophila melanogaster.}, journal = {PLoS genetics}, volume = {12}, number = {4}, pages = {e1005951}, pmid = {27035832}, issn = {1553-7404}, support = {R01 GM045146/GM/NIGMS NIH HHS/United States ; R01 GM45146/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology ; Female ; Genome-Wide Association Study ; Male ; *Recombination, Genetic ; Wolbachia/isolation & purification ; }, abstract = {Meiotic recombination ensures proper chromosome segregation in many sexually reproducing organisms. Despite this crucial function, rates of recombination are highly variable within and between taxa, and the genetic basis of this variation remains poorly understood. Here, we exploit natural variation in the inbred, sequenced lines of the Drosophila melanogaster Genetic Reference Panel (DGRP) to map genetic variants affecting recombination rate. We used a two-step crossing scheme and visible markers to measure rates of recombination in a 33 cM interval on the X chromosome and in a 20.4 cM interval on chromosome 3R for 205 DGRP lines. Though we cannot exclude that some biases exist due to viability effects associated with the visible markers used in this study, we find ~2-fold variation in recombination rate among lines. Interestingly, we further find that recombination rates are uncorrelated between the two chromosomal intervals. We performed a genome-wide association study to identify genetic variants associated with recombination rate in each of the two intervals surveyed. We refined our list of candidate variants and genes associated with recombination rate variation and selected twenty genes for functional assessment. We present strong evidence that five genes are likely to contribute to natural variation in recombination rate in D. melanogaster; these genes lie outside the canonical meiotic recombination pathway. We also find a weak effect of Wolbachia infection on recombination rate and we confirm the interchromosomal effect. Our results highlight the magnitude of population variation in recombination rate present in D. melanogaster and implicate new genetic factors mediating natural variation in this quantitative trait.}, } @article {pmid27033416, year = {2016}, author = {Ramírez-Puebla, ST and Servín-Garcidueñas, LE and Ormeño-Orrillo, E and Vera-Ponce de León, A and Rosenblueth, M and Delaye, L and Martínez, J and Martínez-Romero, E}, title = {A response to Lindsey et al. "Wolbachia pipientis should not be split into multiple species: A response to Ramírez-Puebla et al.".}, journal = {Systematic and applied microbiology}, volume = {39}, number = {3}, pages = {223-225}, doi = {10.1016/j.syapm.2016.03.004}, pmid = {27033416}, issn = {1618-0984}, mesh = {DNA, Bacterial/*genetics ; Genome, Bacterial/*genetics ; *Host-Pathogen Interactions ; Symbiosis ; Wolbachia/*classification/*genetics ; }, } @article {pmid27021523, year = {2016}, author = {Lindsey, ARI and Bordenstein, SR and Newton, ILG and Rasgon, JL}, title = {Wolbachia pipientis should not be split into multiple species: A response to Ramírez-Puebla et al., "Species in Wolbachia? Proposal for the designation of 'Candidatus Wolbachia bourtzisii', 'Candidatus Wolbachia onchocercicola', 'Candidatus Wolbachia blaxteri', 'Candidatus Wolbachia brugii', 'Candidatus Wolbachia taylori', 'Candidatus Wolbachia collembolicola' and 'Candidatus Wolbachia multihospitum' for the different species within Wolbachia supergroups".}, journal = {Systematic and applied microbiology}, volume = {39}, number = {3}, pages = {220-222}, pmid = {27021523}, issn = {1618-0984}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {DNA, Bacterial/*genetics ; Genome, Bacterial/*genetics ; *Host-Pathogen Interactions ; Symbiosis ; Wolbachia/*classification/*genetics ; }, } @article {pmid27021349, year = {2016}, author = {Liu, JN and Wang, TH and Jia, QY and Gao, XH and Wan, H and Sun, WY and Yang, XL and Bao, R and Liu, JZ and Yu, ZJ}, title = {Characterization of the Microbial Communities in the Ant Lion Euroleon coreanus (Okamoto) (Neuroptera: Myrmeleontidae).}, journal = {Neotropical entomology}, volume = {45}, number = {4}, pages = {397-403}, pmid = {27021349}, issn = {1678-8052}, mesh = {Animals ; Bacteria ; China ; Insecta/*microbiology ; Larva/microbiology ; }, abstract = {Euroleon coreanus (Okamoto) is widely distributed in China, and the larval stage can be treated as traditional Chinese medicine. However, the host-bacterium relationship remains unexplored, as there is a lack of knowledge on the microbial community of ant lions. Hence, in the current study, we explored the microbial community of the larval ant lion E. coreanus using Illumina MiSeq sequencing. Results indicated that a total of 10 phyla, 126 genera, and 145 species were characterized from the second instars of E. coreanus, and most of the microbes were classified in the phylum Proteobacteria. Cronobacter muytjensii was the most abundant species characterized in the whole body and gut of E. coreanus, and the unclassified species in the genera Brevundimonas and Lactobacillus were relatively more abundant in the head and carcass. In addition, no Wolbachia-like bacteria were detected, whereas bacteria like Francisella tularensis subsp. Holarctica OSU18 and unclassified Rickettsiella were first identified in ant lion E. coreanus.}, } @article {pmid27018441, year = {2016}, author = {Hubert, J and Stejskal, V and Nesvorna, M and Aulicky, R and Kopecky, J and Erban, T}, title = {Differences in the Bacterial Community of Laboratory and Wild Populations of the Predatory Mite Cheyletus eruditus (Acarina: Cheyletidae) and Bacteria Transmission From Its Prey Acarus siro (Acari: Acaridae).}, journal = {Journal of economic entomology}, volume = {109}, number = {3}, pages = {1450-1457}, doi = {10.1093/jee/tow032}, pmid = {27018441}, issn = {1938-291X}, abstract = {The parthenogenetic predatory mite Cheyletus eruditus (Schrank, 1781) is used for biological control against mite pests produced as CHEYLETIN. Although there is evidence that bacteria are mainly responsible for parthenogeny in several species of predatory mites, the description of association between C. eruditus the specific and parasitic or symbiotic bacteria is still missing. We analyzed the bacterial communities of the predator, C. eruditus , and its prey, Acarus siro L. The 16S rRNA gene was amplified, cloned, and sequenced. The selected bacterial taxa were confirmed by amplification of isolated DNA with taxon-specific primers. The 16S rRNA gene sequences from the predatory and prey mites formed a total of 20 different bacterial taxa. Of these taxa, the predator and prey shared four taxa, six taxa were specific for the predatory, and 10 taxa for the prey mites. Cardinium - and Bartonella -like bacteria were found in both mite species. The reproductive parasite Wolbachia was found only in the predatory mite, and A. siro hosted Solitalea -like (Sphingobacteriales) bacteria that were not detected in C. eruditus . We focused on Cardinium occurrence in the field samples of C. eruditus. Using Cardinium -specific primers, 128 clones were obtained. Cardinium was found in seven field samples of C. eruditus as well as in the laboratory population that was used to produce CHEYLETIN. Phylogenetic analysis of the Cardinium clones identified three separate clusters: two clusters showed high similarity to the Cardinium sequences from astigmatid mites, and one cluster contained only the clones from C. eruditus . Sequences of both Cardinium and Wolbachia were found in the both adults and eggs of C. eruditus , indicating maternal transfer of these endosymbiotic bacteria.}, } @article {pmid27012719, year = {2016}, author = {Wiwatanaratanabutr, I and Zhang, C}, title = {Wolbachia infections in mosquitoes and their predators inhabiting rice field communities in Thailand and China.}, journal = {Acta tropica}, volume = {159}, number = {}, pages = {153-160}, doi = {10.1016/j.actatropica.2016.03.026}, pmid = {27012719}, issn = {1873-6254}, mesh = {Agriculture ; Animals ; China ; Culicidae/*genetics/*microbiology ; DNA, Bacterial/*genetics ; Odonata/*genetics/*microbiology ; Oryza ; Thailand ; Wolbachia/*genetics/*isolation & purification ; }, abstract = {Wolbachia are inherited, endocytoplasmic bacteria that infect a wide range of arthropods. Here is the first systematic report on the study of Wolbachia infection in mosquitoes and their predators from both Thailand and China. In Thailand, 632 mosquito specimens (20 spp.) and 424 insect predators (23 spp.) were collected from the rice agroecosystem, mostly from the Central region, followed by the Northeast, the North and the South and were inhabiting rice fields, wetlands and ditches. In China, 928 mosquitoes (15 spp.) and 149 insect predators (16 spp.) were collected from rice fields along the Weishan Lake in Shandong province. Specimens were classified in the orders Diptera, Coleoptera, Odonata and Hemiptera. Using wsp, ftsZ, 16S rRNA and groE gene amplifications, Wolbachia were detected in 12 mosquito spp. and 6 predator spp. from Thailand and 11 mosquito spp. and 5 predator spp. from China. The relative Wolbachia densities of these species were determined using quantitative real-time PCR. The mosquito, Aedes albopictus, and the predator, Agriocnemis femina, had the highest bacterial densities. These results imply that Wolbachia of supergroup B are distributed throughout these insects, probably via horizontal transmission in rice agroecosystems.}, } @article {pmid27004796, year = {2016}, author = {Dittmer, J and Lesobre, J and Moumen, B and Bouchon, D}, title = {Host origin and tissue microhabitat shaping the microbiota of the terrestrial isopod Armadillidium vulgare.}, journal = {FEMS microbiology ecology}, volume = {92}, number = {5}, pages = {fiw063}, doi = {10.1093/femsec/fiw063}, pmid = {27004796}, issn = {1574-6941}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; Isopoda/*microbiology/physiology ; Microbiota ; RNA, Ribosomal, 16S/genetics ; *Soil Microbiology ; Symbiosis ; Wolbachia/genetics/*isolation & purification/physiology ; }, abstract = {We present the first in-depth investigation of the host-associated microbiota of the terrestrial isopod crustacean Armadillidium vulgare. This species is an important decomposer of organic matter in terrestrial ecosystems and a major model organism for arthropod-Wolbachia symbioses due to its well-characterized association with feminizing Wolbachia 16S rRNA gene pyrotags were used to characterize its bacterial microbiota at multiple levels: (i) in individuals from laboratory lineages and field populations and (ii) in various host tissues. This integrative approach allowed us to reveal an unexpectedly high bacterial diversity, placing this species in the same league as termites in terms of symbiotic diversity. Interestingly, both animal groups belong to the same ecological guild in terrestrial ecosystems. While Wolbachia represented the predominant taxon in infected individuals, it was not the only major player. Together, the most abundant taxa represented a large scope of symbiotic interactions, including bacterial pathogens, a reproductive parasite (Wolbachia) and potential nutritional symbionts. Furthermore, we demonstrate that individuals from different populations harboured distinct bacterial communities, indicating a strong link between the host-associated microbiota and environmental bacteria, possibly due to terrestrial isopod nutritional ecology. Overall, this work highlights the need for more studies of host-microbiota interactions and bacterial diversity in non-insect arthropods.}, } @article {pmid26999590, year = {2016}, author = {Monnin, D and Kremer, N and Berny, C and Henri, H and Dumet, A and Voituron, Y and Desouhant, E and Vavre, F}, title = {Influence of oxidative homeostasis on bacterial density and cost of infection in Drosophila-Wolbachia symbioses.}, journal = {Journal of evolutionary biology}, volume = {29}, number = {6}, pages = {1211-1222}, doi = {10.1111/jeb.12863}, pmid = {26999590}, issn = {1420-9101}, mesh = {Animals ; Drosophila/*physiology ; *Homeostasis ; Oxidation-Reduction ; Phenotype ; *Reactive Oxygen Species ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {The evolution of symbioses along the continuum between parasitism and mutualism can be influenced by the oxidative homeostasis, that is the balance between reactive oxygen species (ROS) and antioxidant molecules. Indeed, ROS can contribute to the host immune defence to regulate symbiont populations, but are also toxic. This interplay between ROS and symbiosis is notably exemplified by recent results in arthropod-Wolbachia interactions. Wolbachia are symbiotic bacteria involved in a wide range of interactions with their arthropods hosts, from facultative, parasitic associations to obligatory, mutualistic ones. In this study, we used Drosophila-Wolbachia associations to determine whether the oxidative homeostasis plays a role in explaining the differences between phenotypically distinct arthropod-Wolbachia symbioses. We used Drosophila lines with different Wolbachia infections and measured the effects of pro-oxidant (paraquat) and antioxidant (glutathione) treatments on the Wolbachia density and the host survival. We show that experimental manipulations of the oxidative homeostasis can reduce the cost of the infection through its effect on Wolbachia density. We discuss the implication of this result from an evolutionary perspective and argue that the oxidative homeostasis could underlie the evolution of tolerance and dependence on Wolbachia.}, } @article {pmid26997548, year = {2016}, author = {Wang, GH and Jia, LY and Xiao, JH and Huang, DW}, title = {Discovery of a new Wolbachia supergroup in cave spider species and the lateral transfer of phage WO among distant hosts.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {41}, number = {}, pages = {1-7}, doi = {10.1016/j.meegid.2016.03.015}, pmid = {26997548}, issn = {1567-7257}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Bacteriophages/classification/*genetics ; Caves ; Chaperonin 60/genetics ; China ; Cytoskeletal Proteins/genetics ; Electron Transport Complex IV/genetics ; Gene Expression ; Gene Transfer, Horizontal ; Host Specificity ; *Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Spiders/*microbiology ; Symbiosis/physiology ; Wolbachia/classification/*genetics/virology ; }, abstract = {Wolbachia are widespread intracellular bacteria infecting the major classes of arthropods and some filarial nematodes. In arthropods, Wolbachia have evolved various intriguing reproductive manipulations, including cytoplasmic incompatibility, parthenogenesis, feminization, and male killing. Sixteen supergroups of Wolbachia have been identified, named A-Q (except G). Though Wolbachia present great diversity in arthropods, spiders, especially cave spiders, are still a poorly surveyed group of Wolbachia hosts. Here, we report a novel Wolbachia supergroup from nine Telema cave spiders (Araneae: Telemidae) based on five molecular markers (16S rRNA, ftsZ, gltA, groEL, and coxA). In addition, phage WO, which was previously reported only in Wolbachia supergroups A, B, and F, infects this new Wolbachia supergroup. We detected a 100% infection rate for phage WO and Wolbachia in Telema species. The phylogenetic trees of phage WO and Wolbachia are not congruent, which suggests that horizontal transfer of phage WO has occurred in these secluded species. Additionally, these data indicate Telema-Wolbachia-phage WO may be a good model for exploring the horizontal transfer history of WO among different host species.}, } @article {pmid26996237, year = {2016}, author = {Sharma, R and Al Jayoussi, G and Tyrer, HE and Gamble, J and Hayward, L and Guimaraes, AF and Davies, J and Waterhouse, D and Cook, DA and Myhill, LJ and Clare, RH and Cassidy, A and Steven, A and Johnston, KL and Ford, L and Turner, JD and Ward, SA and Taylor, MJ}, title = {Minocycline as a re-purposed anti-Wolbachia macrofilaricide: superiority compared with doxycycline regimens in a murine infection model of human lymphatic filariasis.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {23458}, pmid = {26996237}, issn = {2045-2322}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage/pharmacokinetics ; Brugia malayi/drug effects/parasitology ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Doxycycline/administration & dosage/pharmacokinetics ; Elephantiasis, Filarial/parasitology/*prevention & control ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Mice, SCID ; Minocycline/*administration & dosage/pharmacokinetics ; Wolbachia/*drug effects/pathogenicity ; }, abstract = {Lymphatic filariasis and onchocerciasis are parasitic helminth diseases, which cause severe morbidities such as elephantiasis, skin disease and blindness, presenting a major public health burden in endemic communities. The anti-Wolbachia consortium (A·WOL: http://www.a-wol.com/) has identified a number of registered antibiotics that target the endosymbiotic bacterium, Wolbachia, delivering macrofilaricidal activity. Here we use pharmacokinetics/pharmacodynamics (PK/PD) analysis to rationally develop an anti-Wolbachia chemotherapy by linking drug exposure to pharmacological effect. We compare the pharmacokinetics and anti-Wolbachia efficacy in a murine Brugia malayi model of minocycline versus doxycycline. Doxycycline exhibits superior PK in comparison to minocycline resulting in a 3-fold greater exposure in SCID mice. Monte-Carlo simulations confirmed that a bi-daily 25-40 mg/Kg regimen is bioequivalent to a clinically effective 100-200 mg/day dose for these tetracyclines. Pharmacodynamic studies showed that minocycline depletes Wolbachia more effectively than doxycycline (99.51% vs. 90.35%) after 28 day 25 mg/Kg bid regimens with a more potent block in microfilarial production. PK/PD analysis predicts that minocycline would be expected to be 1.7 fold more effective than doxycycline in man despite lower exposure in our infection models. Our findings warrant onward clinical investigations to examine the clinical efficacy of minocycline treatment regimens against lymphatic filariasis and onchocerciasis.}, } @article {pmid26995349, year = {2016}, author = {Vanthournout, B and Hendrickx, F}, title = {Hidden suppression of sex ratio distortion suggests Red queen dynamics between Wolbachia and its dwarf spider host.}, journal = {Journal of evolutionary biology}, volume = {29}, number = {8}, pages = {1488-1494}, doi = {10.1111/jeb.12861}, pmid = {26995349}, issn = {1420-9101}, mesh = {Animals ; Female ; Reproduction ; *Sex Ratio ; *Spiders ; *Symbiosis ; *Wolbachia ; }, abstract = {Genetic conflict theory predicts strong selection for host nuclear factors suppressing endosymbiont effects on reproduction; however, evidence of these suppressors is currently scarce. This can either be caused by a low suppressor evolution rate, or if suppressors originate frequently, by rapid spread and concurrent masking of their activity by silencing the endosymbiont effect. To explore this, we use two populations of a dwarf spider with a similar female bias, caused by a Wolbachia infection. Using inter- and intrapopulation crosses, we determine that one of these populations demonstrates a higher suppressing capability towards Wolbachia despite having a similar population sex ratio. This suggests that spider and endosymbiont are locked in so-called red queen dynamics where, despite continuous coevolution, average fitness remains the same, hence hiding the presence of the suppressor. Finding different suppressor activity in populations that even lack phenotypic differentiation (i.e. similar sex ratio) further supports the hypothesis that suppressors originate often, but are often hidden by their own mode of action by countering endosymbiont effects.}, } @article {pmid26994075, year = {2016}, author = {Molloy, JC and Sommer, U and Viant, MR and Sinkins, SP}, title = {Wolbachia Modulates Lipid Metabolism in Aedes albopictus Mosquito Cells.}, journal = {Applied and environmental microbiology}, volume = {82}, number = {10}, pages = {3109-3120}, pmid = {26994075}, issn = {1098-5336}, support = {MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; BB/H016511/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; WT095121/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; Cell Line ; *Lipid Metabolism ; Lipids/analysis ; Mass Spectrometry ; *Symbiosis ; Wolbachia/growth & development/metabolism/*physiology ; }, abstract = {UNLABELLED: Certain strains of the intracellular endosymbiont Wolbachia can strongly inhibit or block the transmission of viruses such as dengue virus (DENV) by Aedes mosquitoes, and the mechanisms responsible are still not well understood. Direct infusion and liquid chromatography-Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry-based lipidomics analyses were conducted using Aedes albopictus Aa23 cells that were infected with the wMel and wMelPop strains of Wolbachia in comparison to uninfected Aa23-T cells. Substantial shifts in the cellular lipid profile were apparent in the presence of Wolbachia Most significantly, almost all sphingolipid classes were depleted, and some reductions in diacylglycerols and phosphatidylcholines were also observed. These lipid classes have previously been shown to be selectively enriched in DENV-infected mosquito cells, suggesting that Wolbachia may produce a cellular lipid environment that is antagonistic to viral replication. The data improve our understanding of the intracellular interactions between Wolbachia and mosquitoes.

IMPORTANCE: Mosquitoes transmit a variety of important viruses to humans, such as dengue virus and Zika virus. Certain strains of the intracellular bacterial genus called Wolbachia found in or introduced into mosquitoes can block the transmission of viruses, including dengue virus, but the mechanisms responsible are not well understood. We found substantial shifts in the cellular lipid profiles in the presence of these bacteria. Some lipid classes previously shown to be enriched in dengue virus-infected mosquito cells were depleted in the presence of Wolbachia, suggesting that Wolbachia may produce a cellular lipid environment that inhibits mosquito-borne viruses.}, } @article {pmid26991800, year = {2016}, author = {Zhang, F and Li, Q and Chen, X and Huo, Y and Guo, H and Song, Z and Cui, F and Zhang, L and Fang, R}, title = {Roles of the Laodelphax striatellus Down syndrome cell adhesion molecule in Rice stripe virus infection of its insect vector.}, journal = {Insect molecular biology}, volume = {25}, number = {4}, pages = {413-421}, doi = {10.1111/imb.12226}, pmid = {26991800}, issn = {1365-2583}, mesh = {Amino Acid Sequence ; Animals ; Cloning, Molecular ; DNA, Complementary/genetics/metabolism ; Hemiptera/genetics/growth & development/*microbiology/*physiology ; Insect Proteins/chemistry/*genetics/metabolism ; Insect Vectors/genetics/growth & development/microbiology/physiology ; Neural Cell Adhesion Molecules/chemistry/*genetics/metabolism ; Nymph/genetics/growth & development/microbiology/physiology ; Oryza/*virology ; Plant Diseases/*virology ; RNA, Messenger/genetics/metabolism ; Sequence Alignment ; Symbiosis ; Tenuivirus/*physiology ; Wolbachia/physiology ; }, abstract = {The arthropod Down syndrome cell adhesion molecule (Dscam) mediates pathogen-specific recognition via an extensive protein isoform repertoire produced by alternative splicing. To date, most studies have focused on the subsequent pathogen-specific immune response, and few have investigated the entry into cells of viruses or endosymbionts. In the present study, we cloned and characterized the cDNA of Laodelphax striatellus Dscam (LsDscam) and investigated the function of LsDscam in rice stripe virus (RSV) infection and the influence on the endosymbiont Wolbachia. LsDscam displayed a typical Dscam domain architecture, including 10 immunoglobulin (Ig) domains, six fibronectin type III domains, one transmembrane domain and a cytoplasmic tail. Alternative splicing occurred at the N-termini of the Ig2 and Ig3 domains, the complete Ig7 domain, the transmembrane domain and the C-terminus, comprising 10, 51, 35, two and two variable exons, respectively. Potentially LsDscam could encode at least 71 400 unique isoforms and 17 850 types of extracellular regions. LsDscam was expressed in various L. striatellus tissues. Knockdown of LsDscam mRNA via RNA interference decreased the titres of both RSV and Wolbachia, but did not change the numbers of the extracellular symbiotic bacterium Acinetobacter rhizosphaerae. Specific Dscam isoforms may play roles in enhancing the infection of vector-borne viruses or endosymbionts.}, } @article {pmid26990981, year = {2016}, author = {Zhang, D and Lees, RS and Xi, Z and Bourtzis, K and Gilles, JR}, title = {Combining the Sterile Insect Technique with the Incompatible Insect Technique: III-Robust Mating Competitiveness of Irradiated Triple Wolbachia-Infected Aedes albopictus Males under Semi-Field Conditions.}, journal = {PloS one}, volume = {11}, number = {3}, pages = {e0151864}, pmid = {26990981}, issn = {1932-6203}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; R01AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/microbiology/*physiology/radiation effects ; Animals ; Female ; Male ; Mosquito Control/*methods ; Rickettsiaceae Infections/prevention & control/*transmission ; *Sexual Behavior, Animal ; Sterilization, Reproductive ; *Wolbachia ; }, abstract = {Combination of the sterile insect technique with the incompatible insect technique is considered to be a safe approach to control Aedes albopictus populations in the absence of an accurate and scalable sex separation system or genetic sexing strain. Our previous study has shown that the triple Wolbachia-infected Ae. albopictus strain (wAlbA, wAlbB and wPip) was suitable for mass rearing and females could be completely sterilized as pupae with a radiation dose of at least 28 Gy. However, whether this radiation dose can influence the mating competitiveness of the triple infected males was still unknown. In this study we aimed to evaluate the effects of irradiation on the male mating competitiveness of the triple infected strain under laboratory and semi-field conditions. The results herein indicate that irradiation with a lower, female-sterilizing dose has no negative impact on the longevity of triple infected males while a reduced lifespan was observed in the wild type males (wAlbA and wAlbB) irradiated with a higher male-sterilizing dose, in small cages. At different sterile: fertile release ratios in small cages, triple-infected males induced 39.8, 81.6 and 87.8% sterility in a wild type female population at 1:1, 5:1 and 10:1 release ratios, respectively, relative to a fertile control population. Similarly, irradiated triple infected males induced 31.3, 70.5 and 89.3% sterility at 1:1, 5:1 and 10:1 release ratios, respectively, again relative to the fertile control. Under semi-field conditions at a 5:1 release ratio, relative to wild type males, the mean male mating competitiveness index of 28 Gy irradiated triple-infected males was significantly higher than 35 Gy irradiated wild type males, while triple infected males showed no difference in mean mating competitiveness to either irradiated triple-infected or irradiated wild type males. An unexpected difference was also observed in the relative male mating competitiveness of the triple infected strain after irradiation at 28 Gy dose in small vs large cages, with a higher male mating competitiveness index calculated from results of experiments in the large cages. Based on these results, we consider that the male mating performance of the triple infected strain after irradiation at 28 Gy, a dose required for complete female sterility and the avoidance of population replacement, is approximately equal to that of the wild type males under semi-field conditions. Though field evaluation is required, this suggests that the triple infected strain is suitable for irradiation and release as part of a combined SIT-IIT approach to Ae. albopictus control.}, } @article {pmid26989241, year = {2016}, author = {Loreto, EL and Wallau, GL}, title = {Risks of Wolbachia mosquito control.}, journal = {Science (New York, N.Y.)}, volume = {351}, number = {6279}, pages = {1273}, doi = {10.1126/science.351.6279.1273-b}, pmid = {26989241}, issn = {1095-9203}, mesh = {Animals ; Culicidae/drug effects/*microbiology ; Dengue/*prevention & control/transmission ; Insecticides/pharmacology ; Mosquito Control/*methods ; Risk ; *Wolbachia ; }, } @article {pmid26977591, year = {2016}, author = {Wong, ZS and Brownlie, JC and Johnson, KN}, title = {Impact of ERK activation on fly survival and Wolbachia-mediated protection during virus infection.}, journal = {The Journal of general virology}, volume = {97}, number = {6}, pages = {1446-1452}, doi = {10.1099/jgv.0.000456}, pmid = {26977591}, issn = {1465-2099}, mesh = {Animals ; Dicistroviridae/*immunology ; Drosophila/immunology/microbiology/*virology ; *Immunity, Innate ; *MAP Kinase Signaling System ; Survival Analysis ; Virus Diseases/immunology/*veterinary ; Wolbachia/*growth & development ; }, abstract = {Elevated levels of reactive oxygen species (ROS) provide protection against virus-induced mortality in Drosophila. In addition to contributing to oxidative stress, ROS are known to activate a number of signalling pathways including the extracellular signal-regulated kinases (ERK) signalling cascade. It was recently shown that ERK signalling is important for resistance against viral replication and invasion in cultured Drosophila cells and the gut epithelium of adult flies. Here, using a Drosophila loss-of-function ERK (rolled) mutant we demonstrated that ERK is important for fly survival during virus infection. ERK mutant flies subjected to Drosophila C virus (DCV) oral and systemic infection were more susceptible to virus-induced mortality as compared with wild-type flies. We have demonstrated experimentally that ERK activation is important for fly survival during oral and systemic virus infection. Given that elevated ROS correlates with Wolbachia-mediated antiviral protection, we also investigated the involvement of ERK in antiviral protection in flies infected by Wolbachia. The results indicate that ERK activation is increased in the presence of Wolbachia but this does not appear to influence Wolbachia-mediated antiviral protection, at least during systemic infection.}, } @article {pmid26954675, year = {2016}, author = {Gilbert, JA and Medlock, J and Townsend, JP and Aksoy, S and Ndeffo Mbah, M and Galvani, AP}, title = {Determinants of Human African Trypanosomiasis Elimination via Paratransgenesis.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {3}, pages = {e0004465}, pmid = {26954675}, issn = {1935-2735}, support = {1U01AI115648/AI/NIAID NIH HHS/United States ; S10 RR019895/RR/NCRR NIH HHS/United States ; D43 TW007391/TW/FIC NIH HHS/United States ; U01 AI115648/AI/NIAID NIH HHS/United States ; R01AI068932/AI/NIAID NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; 2U01GM087719/GM/NIGMS NIH HHS/United States ; S10 RR029676/RR/NCRR NIH HHS/United States ; U01 GM087719/GM/NIGMS NIH HHS/United States ; }, mesh = {Africa South of the Sahara ; Animals ; Animals, Genetically Modified ; Communicable Disease Control/methods ; *Disease Eradication ; Disease Transmission, Infectious/prevention & control ; Enterobacteriaceae/genetics/*growth & development ; Entomology/methods ; Humans ; Models, Theoretical ; Population Dynamics ; *Symbiosis ; Trypanosoma brucei brucei/*growth & development ; Trypanosomiasis, African/epidemiology/*prevention & control/transmission ; Tsetse Flies/*microbiology ; Wolbachia/*physiology ; }, abstract = {Human African trypanosomiasis (HAT), transmitted by tsetse flies, has historically infected hundreds of thousands of individuals annually in sub-Saharan Africa. Over the last decade, concerted control efforts have reduced reported cases to below 10,000 annually, bringing complete elimination within reach. A potential technology to eliminate HAT involves rendering the flies resistant to trypanosome infection. This approach can be achieved through the introduction of transgenic Sodalis symbiotic bacteria that have been modified to produce a trypanocide, and propagated via Wolbachia symbionts, which confer a reproductive advantage to the paratransgenic tsetse. However, the population dynamics of these symbionts within tsetse flies have not yet been evaluated. Specifically, the key factors that determine the effectiveness of paratransgenesis have yet to be quantified. To identify the impact of these determinants on T.b. gambiense and T.b. rhodesiense transmission, we developed a mathematical model of trypanosome transmission that incorporates tsetse and symbiont population dynamics. We found that fecundity and mortality penalties associated with Wolbachia or recombinant Sodalis colonization, probabilities of vertical transmission, and tsetse migration rates are fundamental to the feasibility of HAT elimination. For example, we determined that HAT elimination could be sustained over 25 years when Wolbachia colonization minimally impacted fecundity or mortality, and when the probability of recombinant Sodalis vertical transmission exceeded 99.9%. We also found that for a narrow range of recombinant Sodalis vertical transmission probability (99.9-90.6% for T.b. gambiense and 99.9-85.8% for T.b. rhodesiense), cumulative HAT incidence was reduced between 30% and 1% for T.b. gambiense and between 21% and 3% for T.b. rhodesiense, although elimination was not predicted. Our findings indicate that fitness and mortality penalties associated with paratransgenic symbionts, as well as tsetse migration rates, are instrumental to HAT elimination, and should be a key focus in the development of paratransgenic symbionts.}, } @article {pmid26943350, year = {2016}, author = {Michaud, C and Chupeau, C and Bech, N and Thierry, M and Sicard, M and Greve, P and Beltran-Bech, S}, title = {Isolation, characterization and PCR multiplexing of microsatellite loci for two sub-species of terrestrial isopod Porcellio dilatatus (Crustacea, Oniscidea).}, journal = {Genetica}, volume = {144}, number = {2}, pages = {223-228}, pmid = {26943350}, issn = {1573-6857}, mesh = {Animals ; Female ; Genetic Markers ; *Genetic Variation ; Genotype ; Isopoda/*genetics/microbiology ; Male ; *Microsatellite Repeats ; Phylogeny ; *Wolbachia ; }, abstract = {Several microsatellite markers have already been developed for different terrestrial isopod species such as Armadillidium vulgare, A. nasatum and Porcellionides pruinosus. In all these species, the endosymbiont Wolbachia has a feminizing effect that generates a female bias in sex ratio and reduces the number of reproductive males. Thus this can potentially decrease the genetic diversity of host populations. However, in some other isopod species, Wolbachia induces cytoplasmic incompatibility (CI); the most commonly described effect of Wolbachia in arthropods. The CI by rendering some crossings incompatible can reduce the gene flow and strengthen genetic differentiation between isopod populations. To date, the influence of Wolbachia inducing CI on population structure of terrestrial isopods has never been investigated. In this study, we developed 10 polymorphic microsatellite markers shared by two sub-species of Porcellio dilatatus. Crossings between the two sub-species are partially incompatible due to two CI-inducing Wolbachia strains. These new microsatellite markers will allow us to investigate the effect of CI on host genetic differentiation in this species complex.}, } @article {pmid26933490, year = {2016}, author = {Grosemans, T and Morris, K and Thomas, WK and Rigaux, A and Moens, T and Derycke, S}, title = {Mitogenomics reveals high synteny and long evolutionary histories of sympatric cryptic nematode species.}, journal = {Ecology and evolution}, volume = {6}, number = {6}, pages = {1854-1870}, pmid = {26933490}, issn = {2045-7758}, abstract = {Species with seemingly identical morphology but with distinct genetic differences are abundant in the marine environment and frequently co-occur in the same habitat. Such cryptic species are typically delineated using a limited number of mitochondrial and/or nuclear marker genes, which do not yield information on gene order and gene content of the genomes under consideration. We used next-generation sequencing to study the composition of the mitochondrial genomes of four sympatrically distributed cryptic species of the Litoditis marina species complex (PmI, PmII, PmIII, and PmIV). The ecology, biology, and natural occurrence of these four species are well known, but the evolutionary processes behind this cryptic speciation remain largely unknown. The gene order of the mitochondrial genomes of the four species was conserved, but differences in genome length, gene length, and codon usage were observed. The atp8 gene was lacking in all four species. Phylogenetic analyses confirm that PmI and PmIV are sister species and that PmIII diverged earliest. The most recent common ancestor of the four cryptic species was estimated to have diverged 16 MYA. Synonymous mutations outnumbered nonsynonymous changes in all protein-encoding genes, with the Complex IV genes (coxI-III) experiencing the strongest purifying selection. Our mitogenomic results show that morphologically similar species can have long evolutionary histories and that PmIII has several differences in genetic makeup compared to the three other species, which may explain why it is better adapted to higher temperatures than the other species.}, } @article {pmid26925368, year = {2016}, author = {Jeffries, CL and Walker, T}, title = {Wolbachia Biocontrol Strategies for Arboviral Diseases and the Potential Influence of Resident Wolbachia Strains in Mosquitoes.}, journal = {Current tropical medicine reports}, volume = {3}, number = {}, pages = {20-25}, pmid = {26925368}, issn = {2196-3045}, support = {101285//Wellcome Trust/United Kingdom ; }, abstract = {Arboviruses transmitted by mosquitoes are a major cause of human disease worldwide. The absence of vaccines and effective vector control strategies has resulted in the need for novel mosquito control strategies. The endosymbiotic bacterium Wolbachia has been proposed to form the basis for an effective mosquito biocontrol strategy. Resident strains of Wolbachia inhibit viral replication in Drosophila fruit flies and induce a reproductive phenotype known as cytoplasmic incompatibility that allows rapid invasion of insect populations. Transinfection of Wolbachia strains into the principle mosquito vector of dengue virus, Stegomyia aegypti, has resulted in dengue-refractory mosquito lines with minimal effects on mosquito fitness. Wolbachia strains have now been established in wild St. aegypti populations through open releases in dengue-endemic countries. In this review, we outline the current state of Wolbachia-based biocontrol strategies for dengue and discuss the potential impact of resident Wolbachia strains for additional target mosquito species that transmit arboviruses.}, } @article {pmid26915691, year = {2016}, author = {Sullivan, W}, title = {The untapped cell biology of neglected tropical diseases.}, journal = {Molecular biology of the cell}, volume = {27}, number = {5}, pages = {739-743}, pmid = {26915691}, issn = {1939-4586}, support = {R01 GM104486/GM/NIGMS NIH HHS/United States ; 83512-443638//PHS HHS/United States ; }, mesh = {Africa/epidemiology ; Asia/epidemiology ; Filariasis/epidemiology/parasitology ; Humans ; Molecular Biology/*methods ; Rabies/epidemiology ; Symbiosis ; Tropical Medicine/economics/*methods ; Wolbachia/physiology ; }, abstract = {The World Health Organization lists a constellation of 17 tropical diseases that afflict approximately one in six individuals on the planet and, until recently, few resources have been devoted to the treatment and eradication of those diseases. They are often referred to as the diseases of the "bottom billion," because they are most prevalent among the poorest individuals in impoverished tropical nations. However, the few studies that have been performed reveal an extraordinary world of molecular and cellular adaptations that facilitate the pathogens' survival in hosts ranging from insects to humans. A compelling case can be made that even a modest investment toward understanding the basic molecular and cell biology of these neglected pathogens has a high probability of yielding exciting new cellular mechanisms and insights into novel ways of combating these diseases.}, } @article {pmid26913026, year = {2016}, author = {Wang, N and Jia, S and Xu, H and Liu, Y and Huang, D}, title = {Multiple Horizontal Transfers of Bacteriophage WO and Host Wolbachia in Fig Wasps in a Closed Community.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {136}, pmid = {26913026}, issn = {1664-302X}, abstract = {Wolbachia-bacteriophage WO is a good model system for studying interactions between bacteria and viruses. Previous surveys of insect hosts have been conducted via sampling from open or semi-open communities; however, no studies have reported the infection patterns of phage WO of insects living in a closed community. Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium habitat for a variety of fig wasp. Therefore, in this study, we performed a thorough survey of Wolbachia and bacteriophage WO infection patterns in a total of 1406 individuals from 23 fig wasps species living on three different fig tree species. The infection rates of Wolbachia and phage WO were 82.6% (19/23) and 39.1% (9/23), respectively. Additionally, phage WO from fig wasps showed strong insect host specificity based on orf7 sequences from fig wasps and 21 other insect species. Probably due to the physical barrier of fig syconium, most phage WO from fig wasps form a specific clade. Phylogenetic analysis showed the absence of congruence between WO and host Wolbachia, WO and insect host, as well as Wolbachia and fig wasps, suggesting that both Wolbachia and phage WO exchanged frequently and independently within the closed syconium. Thus, the infection pattern of bacteriophage WO from fig wasps appeared quite different from that in other insects living outside, although the effect and the transfer routes of phage WO are unclear, which need to be investigated in the future.}, } @article {pmid26896674, year = {2016}, author = {Wu, LH and Hoffmann, AA and Thomson, LJ}, title = {Taiwanese Trichogramma of Asian Corn Borer: Morphology, ITS-2 rDNA Characterization, and Natural Wolbachia Infection.}, journal = {Journal of insect science (Online)}, volume = {16}, number = {1}, pages = {}, pmid = {26896674}, issn = {1536-2442}, mesh = {Animals ; Base Sequence ; DNA, Ribosomal Spacer/*genetics ; Moths/*parasitology ; Pest Control, Biological ; Phylogeny ; Taiwan ; Wasps/anatomy & histology/genetics/*microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Egg parasitoids of the genus Trichogramma are natural enemies of many lepidopteran borers in agricultural areas around the world. It is important to identify the correct species and ideally focus on endemic Trichogramma for pest control in particular crops. In this study, Trichogramma wasps were collected from parasitized eggs of Asian corn borer in Southwestern Taiwan. Three Trichogramma species, Trichogramma ostriniae Pang and Chen, Trichogramma chilonis Ishii, and T. sp. y, were identified based on morphology and the nucleotide sequence of the internal transcribed spacer 2 (ITS-2) region of rDNA. Although T. ostriniae and T. sp. y appear to be morphologically similar, ITS-2 identity between these two taxa is only 89%. Surprisingly, a commercially released Trichogramma colony thought to be T. chilonis possessed 99% identity (ITS-2) with the field T. sp. y individuals. This suggests past contamination leading to subsitution of the laboratory-reared T. chilonis colony by T. sp. y. Natural populations of all three Trichogramma species were found to be infected by a single Wolbachia strain which was identified using a wsp gene sequence.}, } @article {pmid26894435, year = {2016}, author = {Cattel, J and Kaur, R and Gibert, P and Martinez, J and Fraimout, A and Jiggins, F and Andrieux, T and Siozios, S and Anfora, G and Miller, W and Rota-Stabelli, O and Mouton, L}, title = {Correction: Wolbachia in European Populations of the Invasive Pest Drosophila suzukii: Regional Variation in Infection Frequencies.}, journal = {PloS one}, volume = {11}, number = {2}, pages = {e0150050}, pmid = {26894435}, issn = {1932-6203}, } @article {pmid26891349, year = {2016}, author = {Joubert, DA and Walker, T and Carrington, LB and De Bruyne, JT and Kien, DH and Hoang, Nle T and Chau, NV and Iturbe-Ormaetxe, I and Simmons, CP and O'Neill, SL}, title = {Establishment of a Wolbachia Superinfection in Aedes aegypti Mosquitoes as a Potential Approach for Future Resistance Management.}, journal = {PLoS pathogens}, volume = {12}, number = {2}, pages = {e1005434}, pmid = {26891349}, issn = {1553-7374}, support = {101285//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; Dengue/virology ; Dengue Virus ; Female ; Humans ; Insect Vectors/*immunology ; *Pest Control, Biological/methods ; Rickettsiaceae Infections/*microbiology ; Saliva/microbiology ; Superinfection/*microbiology ; Virus Replication ; *Wolbachia ; }, abstract = {Wolbachia pipientis is an endosymbiotic bacterium estimated to chronically infect between 40-75% of all arthropod species. Aedes aegypti, the principle mosquito vector of dengue virus (DENV), is not a natural host of Wolbachia. The transinfection of Wolbachia strains such as wAlbB, wMel and wMelPop-CLA into Ae. aegypti has been shown to significantly reduce the vector competence of this mosquito for a range of human pathogens in the laboratory. This has led to wMel-transinfected Ae. aegypti currently being released in five countries to evaluate its effectiveness to control dengue disease in human populations. Here we describe the generation of a superinfected Ae. aegypti mosquito line simultaneously infected with two avirulent Wolbachia strains, wMel and wAlbB. The line carries a high overall Wolbachia density and tissue localisation of the individual strains is very similar to each respective single infected parental line. The superinfected line induces unidirectional cytoplasmic incompatibility (CI) when crossed to each single infected parental line, suggesting that the superinfection would have the capacity to replace either of the single constituent infections already present in a mosquito population. No significant differences in fitness parameters were observed between the superinfected line and the parental lines under the experimental conditions tested. Finally, the superinfected line blocks DENV replication more efficiently than the single wMel strain when challenged with blood meals from viremic dengue patients. These results suggest that the deployment of superinfections could be used to replace single infections and may represent an effective strategy to help manage potential resistance by DENV to field deployments of single infected strains.}, } @article {pmid26873314, year = {2016}, author = {Greiman, SE and Rikihisa, Y and Cain, J and Vaughan, JA and Tkach, VV}, title = {Germs within Worms: Localization of Neorickettsia sp. within Life Cycle Stages of the Digenean Plagiorchis elegans.}, journal = {Applied and environmental microbiology}, volume = {82}, number = {8}, pages = {2356-2362}, pmid = {26873314}, issn = {1098-5336}, support = {R15 AI092622/AI/NIAID NIH HHS/United States ; R15AI092622/AI/NIAID NIH HHS/United States ; }, mesh = {Animal Structures/microbiology ; Animals ; Helminths/growth & development/*microbiology ; Life Cycle Stages ; Microscopy, Fluorescence ; Neorickettsia/*isolation & purification/*physiology ; *Symbiosis ; Trematoda/growth & development/*microbiology ; }, abstract = {Neorickettsia spp. are bacterial endosymbionts of parasitic flukes (Digenea) that also have the potential to infect and cause disease (e.g., Sennetsu fever) in the vertebrate hosts of the fluke. One of the largest gaps in our knowledge of Neorickettsia biology is the very limited information available regarding the localization of the bacterial endosymbiont within its digenean host. In this study, we used indirect immunofluorescence microscopy to visualize Neorickettsia sp. within several life cycle stages of the digenean Plagiorchis elegans Individual sporocysts, cercariae, metacercariae, and adults of P. elegans naturally infected with Neorickettsia sp. were obtained from our laboratory-maintained life cycle, embedded, sectioned, and prepared for indirect immunofluorescence microscopy using anti-Neorickettsia risticiihorse serum as the primary antibody. Neorickettsiasp. was found within the tegument of sporocysts, throughout cercarial embryos (germ balls) and fully formed cercariae (within the sporocysts), throughout metacercariae, and within the tegument, parenchyma, vitellaria, uteri, testes, cirrus sacs, and eggs of adults. Interestingly, Neorickettsia sp. was not found within the ovarian tissue. This suggests that vertical transmission of Neorickettsia within adult digeneans occurs via the incorporation of infected vitelline cells into the egg rather than direct infection of the ooplasm of the oocyte, as has been described for other bacterial endosymbionts of invertebrates (e.g.,Rickettsia and Wolbachia).}, } @article {pmid26865969, year = {2016}, author = {Enders, LS and Miller, NJ}, title = {Stress-induced changes in abundance differ among obligate and facultative endosymbionts of the soybean aphid.}, journal = {Ecology and evolution}, volume = {6}, number = {3}, pages = {818-829}, pmid = {26865969}, issn = {2045-7758}, abstract = {Bacterial endosymbionts can drive evolutionary novelty by conferring adaptive benefits under adverse environmental conditions. Among aphid species there is growing evidence that symbionts influence tolerance to various forms of stress. However, the extent to which stress inflicted on the aphid host has cascading effects on symbiont community dynamics remains poorly understood. Here we simultaneously quantified the effect of host-plant induced and xenobiotic stress on soybean aphid (Aphis glycines) fitness and relative abundance of its three bacterial symbionts. Exposure to soybean defensive stress (Rag1 gene) and a neurotoxic insecticide (thiamethoxam) substantially reduced aphid composite fitness (survival × reproduction) by 74 ± 10% and 92 ± 2%, respectively, which in turn induced distinctive changes in the endosymbiont microbiota. When challenged by host-plant defenses a 1.4-fold reduction in abundance of the obligate symbiont Buchnera was observed across four aphid clonal lines. Among facultative symbionts of Rag1-stressed aphids, Wolbachia abundance increased twofold and Arsenophonus decreased 1.5-fold. A similar pattern was observed under xenobiotic stress, with Buchnera and Arsenophonus titers decreasing (1.3-fold) and Wolbachia increasing (1.5-fold). Furthermore, variation in aphid virulence to Rag1 was positively correlated with changes in Arsenophonus titers, but not Wolbachia or Buchnera. A single Arsenophonus multi-locus genotype was found among aphid clonal lines, indicating strain diversity is not primarily responsible for correlated host-symbiont stress levels. Overall, our results demonstrate the nature of aphid symbioses can significantly affect the outcome of interactions under stress and suggests general changes in the microbiome can occur across multiple stress types.}, } @article {pmid26862076, year = {2016}, author = {Shokal, U and Yadav, S and Atri, J and Accetta, J and Kenney, E and Banks, K and Katakam, A and Jaenike, J and Eleftherianos, I}, title = {Effects of co-occurring Wolbachia and Spiroplasma endosymbionts on the Drosophila immune response against insect pathogenic and non-pathogenic bacteria.}, journal = {BMC microbiology}, volume = {16}, number = {}, pages = {16}, pmid = {26862076}, issn = {1471-2180}, mesh = {Animals ; Drosophila Proteins/genetics/immunology ; Drosophila melanogaster/*immunology/*microbiology/physiology ; Female ; Male ; Spiroplasma/*physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Symbiotic interactions between microbes and animals are common in nature. Symbiotic organisms are particularly common in insects and, in some cases, they may protect their hosts from pathogenic infections. Wolbachia and Spiroplasma endosymbionts naturally inhabit various insects including Drosophila melanogaster fruit flies. Therefore, this symbiotic association is considered an excellent model to investigate whether endosymbiotic bacteria participate in host immune processes against certain pathogens. Here we have investigated whether the presence of Wolbachia alone or together with Spiroplasma endosymbionts in D. melanogaster adult flies affects the immune response against the virulent insect pathogen Photorhabdus luminescens and against non-pathogenic Escherichia coli bacteria.

RESULTS: We found that D. melanogaster flies carrying no endosymbionts, those carrying both Wolbachia and Spiroplasma, and those containing Wolbachia only had similar survival rates after infection with P. luminescens or Escherichia coli bacteria. However, flies carrying both endosymbionts or Wolbachia only contained higher numbers of E. coli cells at early time-points post infection than flies without endosymbiotic bacteria. Interestingly, flies containing Wolbachia only had lower titers of this endosymbiont upon infection with the pathogen P. luminescens than uninfected flies of the same strain. We further found that the presence of Wolbachia and Spiroplasma in D. melanogaster up-regulated certain immune-related genes upon infection with P. luminescens or E. coli bacteria, but it failed to alter the phagocytic ability of the flies toward E. coli inactive bioparticles.

CONCLUSION: Our results suggest that the presence of Wolbachia and Spiroplasma in D. melanogaster can modulate immune signaling against infection by certain insect pathogenic and non-pathogenic bacteria. Results from such studies are important for understanding the molecular basis of the interactions between endosymbiotic bacteria of insects and exogenous microbes.}, } @article {pmid26859724, year = {2016}, author = {Keroack, CD and Wurster, JI and Decker, CG and Williams, KM and Slatko, BE and Foster, JM and Williams, SA}, title = {Absence of the Filarial Endosymbiont Wolbachia in Seal Heartworm (Acanthocheilonema spirocauda) but Evidence of Ancient Lateral Gene Transfer.}, journal = {The Journal of parasitology}, volume = {102}, number = {3}, pages = {312-318}, doi = {10.1645/15-872}, pmid = {26859724}, issn = {1937-2345}, mesh = {Acanthocheilonema/genetics/*microbiology ; Acanthocheilonemiasis/microbiology/parasitology/*veterinary ; Animals ; Biological Evolution ; Blotting, Western ; DNA Barcoding, Taxonomic ; DNA, Helminth/chemistry/isolation & purification ; Female ; *Gene Transfer, Horizontal/physiology ; Hydroxymethylbilane Synthase/genetics ; Phoca/*parasitology ; Phylogeny ; Polymerase Chain Reaction/methods ; Pseudogenes ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*genetics/immunology/physiology ; }, abstract = {The symbiotic relationship of Wolbachia spp. was first observed in insects and subsequently in many parasitic filarial nematodes. This bacterium is believed to provide metabolic and developmental assistance to filarial parasitic nematodes, although the exact nature of this relationship remains to be fully elucidated. While Wolbachia is present in most filarial nematodes in the family Onchocercidae, it is absent in several disparate species such as the human parasite Loa loa . All tested members of the genus Acanthocheilonema, such as Acanthocheilonema viteae, have been shown to lack Wolbachia. Consistent with this, we show that Wolbachia is absent from the seal heartworm (Acanthocheilonema spirocauda), but lateral gene transfer (LGT) of DNA sequences between Wolbachia and A. spirocauda has occurred, indicating a past evolutionary association. Seal heartworm is an important pathogen of phocid seals and understanding its basic biology is essential for conservation of the host. The findings presented here may allow for the development of future treatments or diagnostics for the disease and also aid in clarification of the complicated nematode-Wolbachia relationship.}, } @article {pmid26856916, year = {2016}, author = {Ye, YH and Carrasco, AM and Dong, Y and Sgrò, CM and McGraw, EA}, title = {The Effect of Temperature on Wolbachia-Mediated Dengue Virus Blocking in Aedes aegypti.}, journal = {The American journal of tropical medicine and hygiene}, volume = {94}, number = {4}, pages = {812-819}, pmid = {26856916}, issn = {1476-1645}, mesh = {Aedes/physiology/*virology ; Animals ; Dengue/prevention & control/transmission ; Dengue Virus/*physiology ; Female ; Humans ; Insect Vectors/microbiology/physiology/virology ; Male ; Pest Control, Biological/*methods ; *Temperature ; Virus Replication/physiology ; Wolbachia/*physiology ; }, abstract = {Dengue fever, caused by dengue virus (DENV), is endemic in more than 100 countries. The lack of effective treatment of patients and the suboptimal efficacies of the tetravalent vaccine in trials highlight the urgent need to develop alternative strategies to lessen the burden of dengue fever.Wolbachia pipientis, an obligate intracellular bacterium, is being developed as a biocontrol strategy against dengue because it limits the replication of the DENV in the mosquito vector,Aedes aegypti However, several recent studies have demonstrated the sensitivity of pathogens, vectors, and their symbionts to temperature. To understand how the tripartite interactions between the mosquito, DENV, and Wolbachia may change under different temperature regimes, we assessed the vector competence and transmission potential of DENV-infected mosquitoes reared at a common laboratory setting of a constant 25°C and at two diurnal temperature settings with mean of 25°C and 28°C and a fluctuating range of 8°C (±4°C). Temperature significantly affected DENV infection rate in the mosquitoes. Furthermore, temperature significantly influenced the proportion of mosquitoes that achieved transmission potential as measured by the presence of virus in the saliva. Regardless of the temperature regimes,Wolbachia significantly and efficiently reduced the proportion of mosquitoes achieving infection and transmission potential across all the temperature regimes studied. This work reinforces the robustness of the Wolbachia biocontrol strategy to field conditions in Cairns, Australia, and suggests that similar studies are required for local mosquito genotypes and field relevant temperatures for emerging field release sites globally.}, } @article {pmid26855259, year = {2016}, author = {Nag, JK and Chahar, D and Shrivastava, N and Gupta, CL and Bajpai, P and Chandra, D and Misra-Bhattacharya, S}, title = {Functional attributes of evolutionary conserved Arg45 of Wolbachia (Brugia malayi) translation initiation factor-1.}, journal = {Future microbiology}, volume = {11}, number = {2}, pages = {195-214}, doi = {10.2217/fmb.15.135}, pmid = {26855259}, issn = {1746-0921}, mesh = {Animals ; *Arginine ; Bacterial Proteins/chemistry/*genetics/*metabolism ; Biological Evolution ; Brugia malayi/microbiology ; DNA/metabolism ; Electrophoretic Mobility Shift Assay ; Humans ; Mutagenesis, Site-Directed ; Phylogeny ; Point Mutation ; Prokaryotic Initiation Factor-1/chemistry/*genetics/*metabolism ; Protein Binding ; RNA/metabolism ; Sequence Alignment ; Wolbachia/*genetics/*metabolism ; }, abstract = {AIM: Wolbachia is a promising antifilarial chemotherapeutic target. Translation initiation factor-1 (Tl IF-1) is an essential factor in prokaryotes. Functional characterization of Wolbachia's novel proteins/enzymes is necessary for the development of adulticidal drugs.

MATERIALS & METHODS: Mutant, Wol Tl IF-1 R45D was constructed by site directed mutagenesis. Fluorimetry and size exclusion chromatography were used to determine the biophysical characteristics. Mobility shift assay and fluorescence resonance energy transfer were used to investigate the functional aspect of Wol Tl IF-1 with its mutant.

RESULTS: Both wild and mutant were in monomeric native conformations. Wild exhibits nonspecific binding with ssRNA/ssDNA fragments under electrostatic conditions and showed annealing and displacement of RNA strands in comparison to mutant.

CONCLUSION: Point mutation impaired RNA chaperone activity of the mutant and its interaction with nucleotides.}, } @article {pmid26851594, year = {2016}, author = {Rana, AK and Kushwaha, S and Singh, PK and Misra-Bhattacharya, S}, title = {Immunological evaluation of an rsmD-like rRNA methyltransferase from Wolbachia endosymbiont of Brugia malayi.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {44}, number = {}, pages = {41-47}, doi = {10.1016/j.cimid.2015.12.001}, pmid = {26851594}, issn = {1878-1667}, mesh = {Animals ; Brugia malayi/physiology ; Cytokines/genetics ; Filariasis/prevention & control ; Immunity, Cellular ; Immunity, Humoral ; Interferon-gamma/genetics ; Interleukin-10/genetics ; Interleukin-4/genetics ; Methyltransferases/genetics/*immunology/isolation & purification ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins/immunology/isolation & purification ; Symbiosis ; Wolbachia/*enzymology/*immunology ; }, abstract = {Wolbachia is a wonderful anti-filarial target with many of its enzymes and surface proteins (WSPs) representing potential drug targets and vaccine candidates. Here we report on the immunologic response of a drug target, rsmD-like rRNA methyltransferase from Wolbachia endosymbiont of Brugia malayi. The recombinant protein generated both humoral and cell-mediated response in BALB/c mice but compromised its immunity. The humoral response was transient and endured barely for six months in mice with or without B. Malayi challenge. In splenocytes of mice, the key humoral immunity mediating cytokine IL4 was lowered (IL4↓) while IFNγ, the major cytokine mediating cellular immunity was decreased along with upregulation of IL10 cytokine (IFNγ↓, IL10↑). The finding here indicates that the enzyme has low immunogenicity and triggers lowering of cytokine level in BALB/c mice. Interestingly the overall immune profile can be summed up with equivalent response generated by WSP or whole Wolbachia.}, } @article {pmid26848744, year = {2016}, author = {Han, T and Lee, W and Lee, S and Park, IG and Park, H}, title = {Reassessment of Species Diversity of the Subfamily Denticollinae (Coleoptera: Elateridae) through DNA Barcoding.}, journal = {PloS one}, volume = {11}, number = {2}, pages = {e0148602}, pmid = {26848744}, issn = {1932-6203}, mesh = {Animals ; Coleoptera/anatomy & histology/*genetics ; *DNA Barcoding, Taxonomic ; Electron Transport Complex IV/chemistry/genetics ; Gene Library ; *Genetic Variation ; Likelihood Functions ; Species Specificity ; }, abstract = {The subfamily Denticollinae is a taxonomically diverse group in the family Elateridae. Denticollinae includes many morphologically similar species and crop pests, as well as many undescribed species at each local fauna. To construct a rapid and reliable identification system for this subfamily, the effectiveness of molecular species identification was assessed based on 421 cytochrome c oxidase subunit I (COI) sequences of 84 morphologically identified species. Among the 84 morphospecies, molecular species identification of 60 species (71.4%) was consistent with their morphological identifications. Six cryptic and/or pseudocryptic species with large genetic divergence (>5%) were confirmed by their sympatric or allopatric distributions. However, 18 species, including a subspecies, had ambiguous genetic distances and shared overlapping intra- and interspecific genetic distances (range: 2.12%-3.67%) suggesting incomplete lineage sorting, introgression of mitochondrial genome, or affection by endosymbionts, such as Wolbachia infection, between species and simple genetic variation within species. In this study, we propose a conservative threshold of 3.6% for convenient molecular operational taxonomic unit (MOTU) identification in the subfamily Denticollinae based on the results of pairwise genetic distances analyses using neighbor-joining, mothur, Automatic Barcode Gap Discovery analysis, and tree-based species delimitation by Poisson Tree Processes analysis. Using the 3.6% threshold, we identified 87 MOTUs and found 8 MOTUs in the interval between 2.5% to 3.5%. Evaluation of MOTUs identified in this range requires integrative species delimitation, including review of morphological and ecological differences as well as sensitive genetic markers. From this study, we confirmed that COI sequence is useful for reassessing species diversity for polymorphic and polytypic species occurring in sympatric and allopatric distributions, and for a single species having an extensively large habitat.}, } @article {pmid26846713, year = {2016}, author = {Schuler, H and Köppler, K and Daxböck-Horvath, S and Rasool, B and Krumböck, S and Schwarz, D and Hoffmeister, TS and Schlick-Steiner, BC and Steiner, FM and Telschow, A and Stauffer, C and Arthofer, W and Riegler, M}, title = {The hitchhiker's guide to Europe: the infection dynamics of an ongoing Wolbachia invasion and mitochondrial selective sweep in Rhagoletis cerasi.}, journal = {Molecular ecology}, volume = {25}, number = {7}, pages = {1595-1609}, pmid = {26846713}, issn = {1365-294X}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Bayes Theorem ; DNA, Mitochondrial/*genetics ; Disease Transmission, Infectious ; Europe ; *Evolution, Molecular ; Gene Frequency ; *Genetics, Population ; Genome, Insect ; Genotype ; Germany ; Haplotypes ; Microsatellite Repeats ; Models, Genetic ; Selection, Genetic ; Sequence Analysis, DNA ; Spatio-Temporal Analysis ; Tephritidae/*genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia is a maternally inherited and ubiquitous endosymbiont of insects. It can hijack host reproduction by manipulations such as cytoplasmic incompatibility (CI) to enhance vertical transmission. Horizontal transmission of Wolbachia can also result in the colonization of new mitochondrial lineages. In this study, we present a 15-year-long survey of Wolbachia in the cherry fruit fly Rhagoletis cerasi across Europe and the spatiotemporal distribution of two prevalent strains, wCer1 and wCer2, and associated mitochondrial haplotypes in Germany. Across most of Europe, populations consisted of either 100% singly (wCer1) infected individuals with haplotype HT1, or 100% doubly (wCer1&2) infected individuals with haplotype HT2, differentiated only by a single nucleotide polymorphism. In central Germany, singly infected populations were surrounded by transitional populations, consisting of both singly and doubly infected individuals, sandwiched between populations fixed for wCer1&2. Populations with fixed infection status showed perfect association of infection and mitochondria, suggesting a recent CI-driven selective sweep of wCer2 linked with HT2. Spatial analysis revealed a range expansion for wCer2 and a large transition zone in which wCer2 splashes appeared to coalesce into doubly infected populations. Unexpectedly, the transition zone contained a large proportion (22%) of wCer1&2 individuals with HT1, suggesting frequent intraspecific horizontal transmission. However, this horizontal transmission did not break the strict association between infection types and haplotypes in populations outside the transition zone, suggesting that this horizontally acquired Wolbachia infection may be transient. Our study provides new insights into the rarely studied Wolbachia invasion dynamics in field populations.}, } @article {pmid26846216, year = {2016}, author = {Chu, CC and Gill, TA and Hoffmann, M and Pelz-Stelinski, KS}, title = {Inter-Population Variability of Endosymbiont Densities in the Asian Citrus Psyllid (Diaphorina citri Kuwayama).}, journal = {Microbial ecology}, volume = {71}, number = {4}, pages = {999-1007}, pmid = {26846216}, issn = {1432-184X}, mesh = {Alphaproteobacteria/*classification/genetics ; Animals ; Bacterial Load ; Citrus/microbiology ; Female ; Florida ; Helicobacter/classification/genetics ; Hemiptera/*microbiology ; Male ; North America ; Real-Time Polymerase Chain Reaction ; Symbiosis ; Wolbachia/*classification/genetics ; }, abstract = {The Asian citrus psyllid (Diaphorina citri Kuwayama) is an insect pest capable of transmitting Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus greening in North America. D. citri also harbors three endosymbionts, Wolbachia, Candidatus Carsonella ruddii, and Candidatus Profftella armatura, which may influence D. citri physiology and fitness. Although genomic researches on these bacteria have been conducted, much remains unclear regarding their ecology and inter-population variability in D. citri. The present work examined the densities of each endosymbiont in adult D. citri sampled from different populations using quantitative PCR. Under field conditions, the densities of all three endosymbionts positively correlated with each other, and they are associated with D. citri gender and locality. In addition, the infection density of CLas also varied across populations. Although an analysis pooling D. citri from different populations showed that CLas-infected individuals tended to have lower endosymbiont densities compared to uninfected individuals, the difference was not significant when the population was included as a factor in the analysis, suggesting that other population-specific factors may have stronger effects on endosymbiont densities. To determine whether there is a genetic basis to the density differences, endosymbiont densities between aged CLas-negative females of two D. citri populations reared under standardized laboratory conditions were compared. Results suggested that inter-population variability in Wolbachia infection density is associated with the genotypes of the endosymbiont or the host. Findings from this work could facilitate understanding of D. citri-bacterial associations that may benefit the development of approaches for managing citrus greening, such as prevention of CLas transmission.}, } @article {pmid26836631, year = {2016}, author = {Rosenfeld, JA and Reeves, D and Brugler, MR and Narechania, A and Simon, S and Durrett, R and Foox, J and Shianna, K and Schatz, MC and Gandara, J and Afshinnekoo, E and Lam, ET and Hastie, AR and Chan, S and Cao, H and Saghbini, M and Kentsis, A and Planet, PJ and Kholodovych, V and Tessler, M and Baker, R and DeSalle, R and Sorkin, LN and Kolokotronis, SO and Siddall, ME and Amato, G and Mason, CE}, title = {Genome assembly and geospatial phylogenomics of the bed bug Cimex lectularius.}, journal = {Nature communications}, volume = {7}, number = {}, pages = {10164}, pmid = {26836631}, issn = {2041-1723}, support = {F31 GM111053/GM/NIGMS NIH HHS/United States ; R01NS076465/NS/NINDS NIH HHS/United States ; P30 CA008748/CA/NCI NIH HHS/United States ; T32 GM083937/GM/NIGMS NIH HHS/United States ; F31GM111053/GM/NIGMS NIH HHS/United States ; R01 GM092706/GM/NIGMS NIH HHS/United States ; R01 NS076465/NS/NINDS NIH HHS/United States ; 1T32GM083937/GM/NIGMS NIH HHS/United States ; R01GM092706/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bedbugs/*genetics ; Blood ; Chromosome Mapping ; Eating ; Female ; Gene Expression Profiling ; *Gene Expression Regulation, Developmental ; Humans ; Life Cycle Stages/*genetics ; Male ; Models, Molecular ; Phylogeny ; Sequence Analysis, RNA ; }, abstract = {The common bed bug (Cimex lectularius) has been a persistent pest of humans for thousands of years, yet the genetic basis of the bed bug's basic biology and adaptation to dense human environments is largely unknown. Here we report the assembly, annotation and phylogenetic mapping of the 697.9-Mb Cimex lectularius genome, with an N50 of 971 kb, using both long and short read technologies. A RNA-seq time course across all five developmental stages and male and female adults generated 36,985 coding and noncoding gene models. The most pronounced change in gene expression during the life cycle occurs after feeding on human blood and included genes from the Wolbachia endosymbiont, which shows a simultaneous and coordinated host/commensal response to haematophagous activity. These data provide a rich genetic resource for mapping activity and density of C. lectularius across human hosts and cities, which can help track, manage and control bed bug infestations.}, } @article {pmid26818097, year = {2016}, author = {Shaikevich, EV and Vinogradova, EB and Bouattour, A and Gouveia de Almeida, AP}, title = {Genetic diversity of Culex pipiens mosquitoes in distinct populations from Europe: contribution of Cx. quinquefasciatus in Mediterranean populations.}, journal = {Parasites & vectors}, volume = {9}, number = {}, pages = {47}, pmid = {26818097}, issn = {1756-3305}, mesh = {Animals ; Base Sequence ; Culex/*genetics/growth & development/microbiology ; DNA, Mitochondrial/chemistry/genetics ; Europe/epidemiology ; Female ; *Genetic Variation ; Haplotypes ; Humans ; Male ; Mediterranean Region/epidemiology ; Microsatellite Repeats/genetics ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Mosquitoes of the Culex pipiens complex are cosmopolitan, and important vectors of neglected tropical diseases, such as arbovirosis and lymphatic filariasis. Among the complex taxa, Cx. pipiens (with two forms pipiens and molestus) and Cx. quinquefasciatus are the most ubiquitous mosquitoes in temperate and tropical regions respectively. Mosquitoes of this taxa lack of morphological differences between females, but have frank behavioral and physiological differences and have different trophic preferences that influence their vectorial status. Hybridization may change the vectorial capacity of these mosquitoes, increasing vector efficiency and medical importance of resulting hybrids.

METHODS: Culex pipiens s.l. from 35 distinct populations were investigated by the study of mtDNA, symbiotic bacterium Wolbachia pipientis, nuclear DNA and flanking region of microsatellite CQ11 polymorphism using PCR with diagnostic primers, RFLP analysis and sequencing.

RESULTS: Six different mitochondrial haplotypes were revealed by sequencing of the cytochrome oxidase subunit I (COI) gene and three different Wolbachia (wPip) groups were identified. A strong association was observed between COI haplotypes/groups, wPip groups and taxa; haplogroup A and infection with wPipII appear to be typical for Cx. pipiens form pipiens, haplotype D and infection with wPipIV for form molestus, while haplogroup E, characteristic of Cx. quinquefasciatus, were correlated with wPipI and found in Cx. pipiens sl. from coastal regions of Southern Europe and Mediterranean region. Analysis of microsatellite locus and nuclear DNA revealed hybrids between Cx. pipiens form pipiens and form molestus, as well as between Cx. pipiens and Cx. quinquefasciatus, in Mediterranean populations, as opposed to Northern Europe. Phylogenetic analysis of COI sequences yielded a tree topology that supported the RFLP analysis with significant bootstrap values for haplotype D and haplogroup E.

CONCLUSIONS: Molecular identification provides the first evidence of the presence of hybrids between Cx. quinquefasciatus and Cx. pipiens as well as cytoplasmic introgression of Cx. quinquefasciatus into Cx. pipiens as a result of hybridization events in coastal regions of Southern Europe and Mediterranean region. Together with observed hybrids between pipiens and molestus forms, these findings point to the presence of hybrids in these areas, with consequent higher potential for disease transmission.}, } @article {pmid26809123, year = {2016}, author = {Muñoz-Muñoz, F and Ramoneda, J and Pagès, N and Pujol, N and Talavera, S}, title = {Is the morphology of Culicoides intersexes parasitized by mermithid nematodes a parasite adaptation? A morphometric approach to Culicoides circumscriptus (Diptera: Ceratopogonidae).}, journal = {Journal of invertebrate pathology}, volume = {135}, number = {}, pages = {1-9}, doi = {10.1016/j.jip.2016.01.008}, pmid = {26809123}, issn = {1096-0805}, mesh = {Adaptation, Physiological ; Analysis of Variance ; Animals ; Arthropod Antennae/anatomy & histology ; Ceratopogonidae/*anatomy & histology/*parasitology ; Disorders of Sex Development ; Female ; Male ; Mermithoidea/*physiology ; Principal Component Analysis ; Wings, Animal/anatomy & histology ; }, abstract = {Mermithidae is a family of endoparasitic nematodes known to cause intersexuality in arthropods. Intersexes of the genus Culicoides parasitized by mermithids have been the object of several studies aiming to describe their particular morphology. Culicoides intersexes are specimens with male genitalia and feminized sexually dimorphic structures, i.e. antennae, mouthparts and wings. To date, these specimens have only been described qualitatively and a quantitative approach supported by statistical analysis is lacking. Here we conduct morphometric analyses of sexually dimorphic structures in a sample of Culicoides circumscriptus that includes 34 intersexes with the aim of describing precisely the intersexual morphology. The morphology of antennae and the mouthparts was studied by multivariate statistical analysis of linear measures, and wing form by implementing geometric morphometrics techniques. While intersex wings proved to have a similar size to male wings, their shape was intermediate between males and females. However, when allometric shape variation was removed, the wing shape of intersexes was almost identical to that of females. The intersex antennae were morphometrically of the female type, especially when size variation was considered. In contrast, the measured mouthparts (the labrum and the third palpal segment) were halfway between males and females, even when body size was considered. Overall, the antennae and the wings showed a higher degree of feminization than the mouthparts. These findings indicate that the degree of feminization depends both on the morphological structure and on body size. Moreover, we propose that the feminization of the wings and antennae has an adaptive meaning for the parasite, which would favor female-like traits in order to access more easily its breeding sites, where the parasite has plenty of new hosts to infect. Female-like antennae would be beneficial to detect these sites, while having female-like wings would favor the host's capacity of dispersal to these sites.}, } @article {pmid26809119, year = {2016}, author = {Cattel, J and Kaur, R and Gibert, P and Martinez, J and Fraimout, A and Jiggins, F and Andrieux, T and Siozios, S and Anfora, G and Miller, W and Rota-Stabelli, O and Mouton, L}, title = {Wolbachia in European Populations of the Invasive Pest Drosophila suzukii: Regional Variation in Infection Frequencies.}, journal = {PloS one}, volume = {11}, number = {1}, pages = {e0147766}, pmid = {26809119}, issn = {1932-6203}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Drosophila/*microbiology ; Europe ; Multilocus Sequence Typing ; North America ; Wolbachia/classification/genetics/*physiology ; }, abstract = {The invasive pest Drosophila suzukii is characterized by a specific fresh-fruit targeting behavior and has quickly become a menace for the fruit economy of newly infested North American and European regions. D. suzukii carries a strain of the endosymbiotic bacterium Wolbachia, named wSuz, which has a low infection frequency and no reproductive manipulation capabilities in American populations of D. suzukii. To further understand the nature of wSuz biology and assess its utility as a tool for controlling this pest's populations, we investigated the prevalence of Wolbachia in 23 European D. suzukii populations, and compared our results with those available in American populations. Our data showed a highly variable infection frequency with a mean prevalence of 46%, which is significantly higher than the 17% found in American populations. Based on Multilocus Sequence Typing analysis, a single wSuz strain was diagnosed in all European populations of D. suzukii. In agreement with American data, we found no evidence of cytoplasmic incompatibility induced by wSuz. These findings raise two questions: a) why Wolbachia is maintained in field populations of D. suzukii and b) what are the selective forces responsible for the variation in prevalence within populations, particularly between European and American continents? Our results provide new insights into the D. suzukii-Wolbachia association and highlight regional variations that await further investigation and that should be taken into account for using Wolbachia-based pest management programs.}, } @article {pmid26806463, year = {2016}, author = {Sharma, OP and Kumar, MS}, title = {Essential proteins and possible therapeutic targets of Wolbachia endosymbiont and development of FiloBase--a comprehensive drug target database for Lymphatic filariasis.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {19842}, pmid = {26806463}, issn = {2045-2322}, mesh = {Animals ; Bacterial Proteins/*biosynthesis/genetics ; Brugia malayi/microbiology/pathogenicity ; Databases, Genetic ; Elephantiasis, Filarial/drug therapy/*genetics/parasitology ; *Expressed Sequence Tags ; Gene Expression Regulation/genetics ; Genome, Bacterial ; Host-Pathogen Interactions/genetics ; Humans ; Proteome/drug effects/genetics ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Lymphatic filariasis (Lf) is one of the oldest and most debilitating tropical diseases. Millions of people are suffering from this prevalent disease. It is estimated to infect over 120 million people in at least 80 nations of the world through the tropical and subtropical regions. More than one billion people are in danger of getting affected with this life-threatening disease. Several studies were suggested its emerging limitations and resistance towards the available drugs and therapeutic targets for Lf. Therefore, better medicine and drug targets are in demand. We took an initiative to identify the essential proteins of Wolbachia endosymbiont of Brugia malayi, which are indispensable for their survival and non-homologous to human host proteins. In this current study, we have used proteome subtractive approach to screen the possible therapeutic targets for wBm. In addition, numerous literatures were mined in the hunt for potential drug targets, drugs, epitopes, crystal structures, and expressed sequence tag (EST) sequences for filarial causing nematodes. Data obtained from our study were presented in a user friendly database named FiloBase. We hope that information stored in this database may be used for further research and drug development process against filariasis. URL: http://filobase.bicpu.edu.in.}, } @article {pmid26801597, year = {2016}, author = {Montoya-Alonso, JA and Carretón, E and Morchón, R and Silveira-Viera, L and Falcón, Y and Simón, F}, title = {The impact of the climate on the epidemiology of Dirofilaria immitis in the pet population of the Canary Islands.}, journal = {Veterinary parasitology}, volume = {216}, number = {}, pages = {66-71}, doi = {10.1016/j.vetpar.2015.12.005}, pmid = {26801597}, issn = {1873-2550}, mesh = {Age Distribution ; Animals ; Antibodies, Bacterial/blood ; Antibodies, Helminth/blood ; Antigens, Helminth/blood ; Cat Diseases/*epidemiology/parasitology ; Cats ; *Climate ; Dirofilaria immitis/immunology/physiology ; Dirofilariasis/*epidemiology ; Dog Diseases/*epidemiology/parasitology ; Dogs ; Female ; Male ; Pets/*parasitology ; Prevalence ; Seroepidemiologic Studies ; Sex Distribution ; Spain/epidemiology ; Wolbachia/immunology ; }, abstract = {Cardiopulmonary dirofilariosis (heartworm) is a zoonotic vector borne disease caused by Dirofilaria immitis which affects domestic dogs and cats. Two of the seven Canary Islands are historically hyperendemic areas of dirofilariosis, although no epidemiological study has ever been carried out which includes the other islands. The aim of the study was to complete the epidemiological status of cardiopulmonary dirofilariosis in the canine and feline population throughout all the Canary Islands. 1643 client-owned dogs and 707 client-owned cats were tested for D. immitis antigens (dogs), and anti-D. immitis and anti-Wolbachia antibodies (cats). The prevalence of canine dirofilariosis in the Canary Islands was 15.7%, and the seroprevalence of feline dirofilariosis was 18.1%. A remarkable disparity was found when evaluating the results by island separately, which ranged from from 0% in Lanzarote and El Hierro, low prevalences and seroprevalences in Fuerteventura (1.8% and 2.5% in dogs and cats, respectively), to higher prevalences on the other 4 islands; ranging between 15.7% (dogs) and 14.3% (cats) in La Palma 22.5% (dogs) and 24.1% (cats) in Tenerife. In addition, prevalences and seroprevalences were very variable within each island, these differences being associated to local climate conditions. The distribution and prevalence of dirofilariosis in the Canary Islands is heterogeneous and related to climate, demographic factors and management of pets in the studied areas. Dirofilariosis remains hyperendemic in 4 of the 7 Islands. Since D. immitis is a zoonosis, veterinary and health authorities should be aware of the current prevalence and seroprevalence of animal dirofilariosis. The results show the need for awareness raising campaigns to promote the implementation of prophylactic measures in pets, in order to achieve a decrease in the prevalence of animal dirofilariosis in the Canary Islands.}, } @article {pmid26798263, year = {2015}, author = {Augustinos, AA and Drosopoulou, E and Gariou-Papalexiou, A and Asimakis, ED and Cáceres, C and Tsiamis, G and Bourtzis, K and Penelope Mavragani-Tsipidou, and Zacharopoulou, A}, title = {Cytogenetic and symbiont analysis of five members of the B. dorsalis complex (Diptera, Tephritidae): no evidence of chromosomal or symbiont-based speciation events.}, journal = {ZooKeys}, volume = {}, number = {540}, pages = {273-298}, pmid = {26798263}, issn = {1313-2989}, abstract = {The Bactrocera dorsalis species complex, currently comprising about 90 entities has received much attention. During the last decades, considerable effort has been devoted to delimiting the species of the complex. This information is of great importance for agriculture and world trade, since the complex harbours several pest species of major economic importance and other species that could evolve into global threats. Speciation in Diptera is usually accompanied by chromosomal rearrangements, particularly inversions that are assumed to reduce/eliminate gene flow. Other candidates currently receiving much attention regarding their possible involvement in speciation are reproductive symbionts, such as Wolbachia, Spiroplasma, Arsenophonus, Rickettsia and Cardinium. Such symbionts tend to spread quickly through natural populations and can cause a variety of phenotypes that promote pre-mating and/or post-mating isolation and, in addition, can affect the biology, physiology, ecology and evolution of their insect hosts in various ways. Considering all these aspects, we present: (a) a summary of the recently gained knowledge on the cytogenetics of five members of the Bactrocera dorsalis complex, namely Bactrocera dorsalis s.s., Bactrocera invadens, Bactrocera philippinensis, Bactrocera papayae and Bactrocera carambolae, supplemented by additional data from a Bactrocera dorsalis s.s. colony from China, as well as by a cytogenetic comparison between the dorsalis complex and the genetically close species, Bactrocera tryoni, and, (b) a reproductive symbiont screening of 18 different colonized populations of these five taxa. Our analysis did not reveal any chromosomal rearrangements that could differentiate among them. Moreover, screening for reproductive symbionts was negative for all colonies derived from different geographic origins and/or hosts. There are many different factors that can lead to speciation, and our data do not support chromosomal and/or symbiotic-based speciation phenomena in the taxa under study.}, } @article {pmid26783021, year = {2017}, author = {Silvestre-Ferreira, AC and Vieira, L and Vilhena, H and Cerón, JJ and Tvarijonaviciute, A and Montoya-Alonso, JA and Carretón, E and Pastor, J}, title = {Serum acute phase proteins in Dirofilaria immitis and Wolbachia seropositive cats.}, journal = {Journal of feline medicine and surgery}, volume = {19}, number = {6}, pages = {693-696}, doi = {10.1177/1098612X15625435}, pmid = {26783021}, issn = {1532-2750}, mesh = {Acute-Phase Proteins/*analysis ; Animals ; Antibodies, Bacterial/*blood ; Cat Diseases/blood/*diagnosis ; Cats ; Dirofilaria immitis/*immunology ; Dirofilariasis/blood/*diagnosis ; Female ; Male ; Predictive Value of Tests ; Serologic Tests/veterinary ; Wolbachia/*immunology ; }, abstract = {Objectives The aim of this study was to characterise the response of acute phase proteins (APPs) in cats seropositive for Dirofilaria immitis and to its endosymbiont bacterium Wolbachia. Methods The APPs serum amyloid A (SAA), haptoglobin (Hp) and ceruloplasmin (Cp) were measured in 25 seropositive cats and in 16 healthy seronegative cats. Results SAA and Cp concentrations were significantly higher in animals with D immitis seropositivity that exhibited clinical signs related to the disease, and Hp was elevated in all D immitis-seropositive animals. There was no significant correlation between APPs and D immitis or Wolbachia species antibody titres. Conclusions and relevance An association between feline seropositivity to D immitis and APP response was demonstrated. Increases in serum SAA and Cp concentrations were related to D immitis-associated clinical signs, whereas Hp increased in all seropositive animals.}, } @article {pmid26776329, year = {2016}, author = {Caragata, EP and Dutra, HLC and Moreira, LA}, title = {Exploiting Intimate Relationships: Controlling Mosquito-Transmitted Disease with Wolbachia.}, journal = {Trends in parasitology}, volume = {32}, number = {3}, pages = {207-218}, doi = {10.1016/j.pt.2015.10.011}, pmid = {26776329}, issn = {1471-5007}, mesh = {Animals ; Culicidae/*microbiology ; Insect Vectors/*microbiology ; Malaria/*prevention & control ; Mosquito Control/*trends ; Wolbachia/*physiology ; }, abstract = {Mosquito-transmitted diseases impose a growing burden on human health, and current control strategies have proven insufficient to stem the tide. The bacterium Wolbachia is a novel and promising form of control for mosquito-transmitted disease. It manipulates host biology, restricts infection with dengue and other pathogens, and alters host reproduction to promote rapid spread in the field. In this review, we examine how the intimate and diverse relationships formed between Wolbachia and their mosquito hosts can be exploited for disease control purposes. We consider these relationships in the context of recent developments, including successful field trials with Wolbachia-infected mosquitoes to combat dengue, and new Wolbachia infections in key malaria vectors, which have enhanced the disease control prospects of this unique bacterium.}, } @article {pmid26774684, year = {2016}, author = {Bourtzis, K and Lees, RS and Hendrichs, J and Vreysen, MJ}, title = {More than one rabbit out of the hat: Radiation, transgenic and symbiont-based approaches for sustainable management of mosquito and tsetse fly populations.}, journal = {Acta tropica}, volume = {157}, number = {}, pages = {115-130}, doi = {10.1016/j.actatropica.2016.01.009}, pmid = {26774684}, issn = {1873-6254}, mesh = {Aedes/microbiology/virology ; Animals ; *Animals, Genetically Modified ; Chikungunya Fever/*prevention & control ; Dengue/*prevention & control ; Disease Management ; Disease Vectors ; Humans ; *Insecticides ; *Pest Control, Biological ; Trypanosomiasis, African/*prevention & control ; Tsetse Flies/microbiology/virology ; Zika Virus Infection/*prevention & control ; }, abstract = {Mosquitoes (Diptera: Culicidae) and tsetse flies (Diptera: Glossinidae) are bloodsucking vectors of human and animal pathogens. Mosquito-borne diseases (malaria, filariasis, dengue, zika, and chikungunya) cause severe mortality and morbidity annually, and tsetse fly-borne diseases (African trypanosomes causing sleeping sickness in humans and nagana in livestock) cost Sub-Saharan Africa an estimated US$ 4750 million annually. Current reliance on insecticides for vector control is unsustainable: due to increasing insecticide resistance and growing concerns about health and environmental impacts of chemical control there is a growing need for novel, effective and safe biologically-based methods that are more sustainable. The integration of the sterile insect technique has proven successful to manage crop pests and disease vectors, particularly tsetse flies, and is likely to prove effective against mosquito vectors, particularly once sex-separation methods are improved. Transgenic and symbiont-based approaches are in development, and more advanced in (particularly Aedes) mosquitoes than in tsetse flies; however, issues around stability, sustainability and biosecurity have to be addressed, especially when considering population replacement approaches. Regulatory issues and those relating to intellectual property and economic cost of application must also be overcome. Standardised methods to assess insect quality are required to compare and predict efficacy of the different approaches. Different combinations of these three approaches could be integrated to maximise their benefits, and all have the potential to be used in tsetse and mosquito area-wide integrated pest management programmes.}, } @article {pmid26771308, year = {2016}, author = {Ehounoud, CB and Yao, KP and Dahmani, M and Achi, YL and Amanzougaghene, N and Kacou N'Douba, A and N'Guessan, JD and Raoult, D and Fenollar, F and Mediannikov, O}, title = {Multiple Pathogens Including Potential New Species in Tick Vectors in Côte d'Ivoire.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {1}, pages = {e0004367}, pmid = {26771308}, issn = {1935-2735}, mesh = {Anaplasma/genetics/isolation & purification ; Animals ; Arachnid Vectors/classification/genetics/*microbiology ; Borrelia/genetics/isolation & purification ; Cote d'Ivoire ; Ehrlichia/genetics/isolation & purification ; Female ; Male ; Phylogeny ; Rickettsia/genetics/isolation & purification ; Ticks/classification/genetics/*microbiology ; }, abstract = {BACKGROUND: Our study aimed to assess the presence of different pathogens in ticks collected in two regions in Côte d'Ivoire.

Real-time PCR and standard PCR assays coupled to sequencing were used. Three hundred and seventy eight (378) ticks (170 Amblyomma variegatum, 161 Rhipicepalus microplus, 3 Rhipicephalus senegalensis, 27 Hyalomma truncatum, 16 Hyalomma marginatum rufipes, and 1 Hyalomma impressum) were identified and analyzed. We identified as pathogenic bacteria, Rickettsia africae in Am. variegatum (90%), Rh. microplus (10%) and Hyalomma spp. (9%), Rickettsia aeschlimannii in Hyalomma spp. (23%), Rickettsia massiliae in Rh. senegalensis (33%) as well as Coxiella burnetii in 0.2%, Borrelia sp. in 0.2%, Anaplasma centrale in 0.2%, Anaplasma marginale in 0.5%, and Ehrlichia ruminantium in 0.5% of all ticks. Potential new species of Borrelia, Anaplasma, and Wolbachia were detected. Candidatus Borrelia africana and Candidatus Borrelia ivorensis (detected in three ticks) are phylogenetically distant from both the relapsing fever group and Lyme disease group borreliae; both were detected in Am. variegatum. Four new genotypes of bacteria from the Anaplasmataceae family were identified, namely Candidatus Anaplasma ivorensis (detected in three ticks), Candidatus Ehrlichia urmitei (in nine ticks), Candidatus Ehrlichia rustica (in four ticks), and Candidatus Wolbachia ivorensis (in one tick).

CONCLUSIONS/SIGNIFICANCE: For the first time, we demonstrate the presence of different pathogens such as R. aeschlimannii, C. burnetii, Borrelia sp., A. centrale, A. marginale, and E. ruminantium in ticks in Côte d'Ivoire as well as potential new species of unknown pathogenicity.}, } @article {pmid26771164, year = {2016}, author = {Abegunde, AT and Ahuja, RM and Okafor, NJ}, title = {Doxycycline plus ivermectin versus ivermectin alone for treatment of patients with onchocerciasis.}, journal = {The Cochrane database of systematic reviews}, volume = {2016}, number = {1}, pages = {CD011146}, pmid = {26771164}, issn = {1469-493X}, support = {U01 EY020522/EY/NEI NIH HHS/United States ; 1 U01 EY020522/EY/NEI NIH HHS/United States ; //Department of Health/United Kingdom ; }, mesh = {Doxycycline/adverse effects/*therapeutic use ; Drug Therapy, Combination ; Filaricides/adverse effects/*therapeutic use ; Humans ; Ivermectin/adverse effects/*therapeutic use ; Onchocerciasis/*drug therapy/prevention & control ; Onchocerciasis, Ocular/drug therapy/prevention & control ; Randomized Controlled Trials as Topic ; Vision Disorders/prevention & control ; }, abstract = {BACKGROUND: Onchocerciasis, also known as "river blindness," is a parasitic disease that is caused by infection from the filarial nematode (roundworm), Onchocerca volvulus. Nematodes are transmitted from person to person by blackflies of the Simulium genus, which usually breed in fast flowing streams and rivers. The disease is the second leading infectious cause of blindness in endemic areas.Ivermectin (a microfilaricide) is widely distributed to endemic populations for prevention and treatment of onchocerciasis. Doxycycline, an antibiotic, targets Wolbachia organisms that are crucial to the survival of adult onchocerca (macrofilaricide). Combined treatment with both drugs is believed to cause direct microfilarial death by ivermectin and indirect macrofilarial death by doxycycline. Long-term reduction in the numbers of microfilaria in the skin and eyes and in the numbers of adult worms in the body has the potential to reduce the transmission and occurrence of onchocercal eye disease.

OBJECTIVES: The primary aim of this review was to assess the effectiveness of doxycycline plus ivermectin versus ivermectin alone for prevention and treatment of onchocerciasis. The secondary aim was to assess the effectiveness of doxycycline plus ivermectin versus ivermectin alone for prevention and treatment of onchocercal ocular lesions in communities co-endemic for onchocerciasis and Loa loa (loiasis) infection.

SEARCH METHODS: We searched CENTRAL (which contains the Cochrane Eyes and Vision Trials Register) (Issue 7, 2015), Ovid MEDLINE, Ovid MEDLINE In-Process and Other Non-Indexed Citations, Ovid MEDLINE Daily, Ovid OLDMEDLINE (January 1946 to July 2015), EMBASE (January 1980 to July 2015), PubMed (1948 to July 2015), Latin American and Caribbean Health Sciences Literature Database (LILACS) (1982 to July 2015), the metaRegister of Controlled Trials (mRCT) (www.controlled-trials.com) (last searched 1 July 2014), ClinicalTrials.gov (www.clinicaltrials.gov) and the World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) (www.who.int/ictrp/search/en). We did not use any date or language restrictions in the electronic search for trials. We last searched the electronic databases on 15 July 2015.

SELECTION CRITERIA: We included randomized controlled trials (RCTs) that had compared doxycycline plus ivermectin versus ivermectin alone. Participants with or without one or more characteristic signs of ocular onchocerciasis resided in communities where onchocerciasis was endemic.

DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial eligibility and extracted data. We used standard methodological procedures as expected by Cochrane.

MAIN RESULTS: We identified three RCTs including a total of 466 participants with a diagnosis of onchocerciasis. All trials compared doxycycline plus ivermectin versus ivermectin alone. One study investigated improvement in visual impairment at six-month follow-up; the other two studies measured microfilarial loads in skin snips to assess sustained effects of treatment at follow-up of 21 months or longer. The studies were conducted at various centers across three countries (Cameroon, Ghana, and Liberia). We judged all studies to be at overall high risk of bias because of inadequate randomization and lack of masking (one study), missing data (two studies), and selective outcome reporting (three studies).Only one study measured visual outcomes. This study reported uncertainty about the difference in the proportion of participants with improvement in visual impairment at six-month follow-up for doxycycline plus ivermectin compared with ivermectin alone (risk ratio (RR) 1.06, 95% confidence interval (95% CI) 0.80 to 1.39; 240 participants; very low-quality evidence). No participant in either group showed improvement in optic atrophy, chorioretinitis, or sclerosing keratitis at six-month follow-up. More participants in the doxycycline plus ivermectin group than in the ivermectin alone group showed improvement in iridocyclitis (RR 1.24, 95% CI 0.69 to 2.22) and punctate keratitis (RR 1.43, 95% CI 1.02 to 2.00) at six-month follow-up; however, we graded these results as very low quality.Two studies reported that a six-week course of doxycycline may result in Wolbachia depletion and macrofilaricidal and sterilizing activities in female Onchocerca worms; however, no analysis was possible because data were missing and incomplete (graded evidence as very low quality). Adverse events were reported in 16 of 135 (12%) participants in one of these studies and included itching, headaches, body pains, and vertigo; no difference between treatment groups was reported for any adverse event. The second study reported that one (1.3%) participant in the doxycycline plus ivermectin group had bloody diarrhea after treatment was initiated.

AUTHORS' CONCLUSIONS: Available evidence on the effectiveness of doxycycline plus ivermectin compared with ivermectin alone in preventing and treating onchocerciasis is unclear. Limited evidence of very low quality from two studies indicates that a six-week course of doxycycline followed by ivermectin may result in more frequent macrofilaricidal and microfilaricidal activity and sterilization of female adult Onchocerca compared with ivermectin alone; however, effects on vision-related outcomes are uncertain. Future studies should consider the effectiveness of treatments in preventing visual acuity and visual field loss and their effects on anterior and posterior segment lesions, particularly chorioretinitis. These studies should report outcomes in a uniform and consistent manner at follow-up of three years or longer to allow detection of meaningful changes in vision-related outcomes.}, } @article {pmid26765951, year = {2016}, author = {Atyame, CM and Labbé, P and Lebon, C and Weill, M and Moretti, R and Marini, F and Gouagna, LC and Calvitti, M and Tortosa, P}, title = {Comparison of Irradiation and Wolbachia Based Approaches for Sterile-Male Strategies Targeting Aedes albopictus.}, journal = {PloS one}, volume = {11}, number = {1}, pages = {e0146834}, pmid = {26765951}, issn = {1932-6203}, mesh = {Aedes/*microbiology/*radiation effects ; Animals ; Female ; Humans ; Male ; Mosquito Control/*methods ; *Radiation, Ionizing ; *Wolbachia ; }, abstract = {The global expansion of Aedes albopictus together with the absence of vaccines for most of the arboviruses transmitted by this mosquito has stimulated the development of sterile-male strategies aiming at controlling disease transmission through the suppression of natural vector populations. In this context, two environmentally friendly control strategies, namely the Sterile Insect Technique (SIT) and the Wolbachia-based Incompatible Insect Technique (IIT) are currently being developed in several laboratories worldwide. So far however, there is a lack of comparative assessment of these strategies under the same controlled conditions. Here, we compared the mating capacities, i.e. insemination capacity, sterilization capacity and mating competitiveness of irradiated (35 Gy) and incompatible Ae. albopictus males at different ages and ratios under laboratory controlled conditions. Our data show that there was no significant difference in insemination capacity of irradiated and incompatible males, both male types showing lower capacities than untreated males at 1 day but recovering full capacity within 5 days following emergence. Regarding mating competitiveness trials, a global observed trend is that incompatible males tend to induce a lower hatching rate than irradiated males in cage controlled confrontations. More specifically, incompatible males were found more competitive than irradiated males in 5:1 ratio regardless of age, while irradiated males were only found more competitive than incompatible males in the 1:1 ratio at 10 days old. Overall, under the tested conditions, IIT seemed to be slightly more effective than SIT. However, considering that a single strategy will likely not be adapted to all environments, our data stimulates the need for comparative assessments of distinct strategies in up-scaled conditions in order to identify the most suitable and safe sterilizing technology to be implemented in a specific environmental setting and to identify the parameters requiring fine tuning in order to reach optimal release conditions.}, } @article {pmid26762514, year = {2016}, author = {Muturi, EJ and Kim, CH and Bara, J and Bach, EM and Siddappaji, MH}, title = {Culex pipiens and Culex restuans mosquitoes harbor distinct microbiota dominated by few bacterial taxa.}, journal = {Parasites & vectors}, volume = {9}, number = {}, pages = {18}, pmid = {26762514}, issn = {1756-3305}, mesh = {Animals ; Base Sequence ; Culex/classification/*microbiology ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Female ; Humans ; Illinois ; Insect Vectors/*microbiology ; Microbiota/*genetics ; Molecular Sequence Data ; Mosquito Control ; Sequence Analysis, DNA ; }, abstract = {BACKGROUND: Mosquitoes host diverse microbial communities that influence many aspects of their biology including reproduction, digestion, and ability to transmit pathogens. Unraveling the composition, structure, and function of these microbiota can provide new opportunities for exploiting microbial function for mosquito-borne disease control.

METHODS: MiSeq® sequencing of 16S rRNA gene amplicons was used to characterize the microbiota of adult females of Culex pipiens L. and Cx. restuans Theobald collected from nine study sites in central Illinois.

RESULTS: Out of 195 bacterial OTUs that were identified, 86 were shared between the two mosquito species while 16 and 93 OTUs were unique to Cx. pipiens and Cx. restuans, respectively. The composition and structure of microbial communities differed significantly between the two mosquito species with Cx. restuans hosting a more diverse bacterial community compared to Cx. pipiens. Wolbachia (OTU836919) was the dominant bacterial species in Cx. pipiens accounting for 91% of total microbiota while Sphingomonas (OTU817982) was the dominant bacterial species in Cx. restuans accounting for 31% of total microbiota. Only 3 and 6 OTUs occurred in over 60% of individuals in Cx. pipiens and Cx. restuans, respectively. There was little effect of study site on bacterial community structure of either mosquito species.

CONCLUSION: These results suggest that the two mosquito species support distinct microbial communities that are sparsely distributed between individuals. These findings will allow investigations of the role of identified microbiota on the spatial and temporal heterogeneity in WNV transmission and their potential application in disease control.}, } @article {pmid26747442, year = {2016}, author = {Larson, MA and Nalbantoglu, U and Sayood, K and Zentz, EB and Cer, RZ and Iwen, PC and Francesconi, SC and Bishop-Lilly, KA and Mokashi, VP and Sjöstedt, A and Hinrichs, SH}, title = {Reclassification of Wolbachia persica as Francisella persica comb. nov. and emended description of the family Francisellaceae.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {66}, number = {3}, pages = {1200-1205}, doi = {10.1099/ijsem.0.000855}, pmid = {26747442}, issn = {1466-5034}, mesh = {Bacterial Typing Techniques ; Base Composition ; DNA, Bacterial/genetics ; Francisella/*classification ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/classification ; }, abstract = {The taxonomic status of the bacterium Wolbachia persica is described, and based on the evidence presented, transfer of this species to the genus Francisella as Francisella persica comb. nov. is proposed. This reclassification is supported by data generated from genomic comparisons of W. persica ATCC VR-331T (= FSC845T = DSM 101678T) to other near neighbours, including Francisella tularensis subsp. novicida. The full-length 16S rRNA gene sequence of strain ATCC VR-331T had 98.5 % nucleotide identity to the cognate gene in F. tularensis, with the highest similarity to subspecies novicida. Phylogenetic trees of full-length 16S rRNA gene, gyrA and recA sequences from species of the genera Wolbachia (class Alphaproteobacteria) and Francisella (class Gammaproteobacteria) indicated that W. persica ATCC VR-331T was most closely related to members of the genus Francisella and not Wolbachia. Local collinear blocks within the chromosome of strain ATCC VR-331T had considerable similarity with F. tularensis subsp. novicida, but not with any Wolbachia strain. The genomes of strain ATCC VR-331T and F. tularensis subsp. novicida Utah 112T (= ATCC 15482T) contained an average nucleotide identity mean of 88.72 % and median of 89.18 %. Importantly, the genome of strain ATCC VR-331T contained one Francisella Pathogenicity Island, similar to F. tularensis subsp. novicida, as well as the Francisella-specific gene fopA1 and F. tularensis-specific genes fopA2 and lpnA (also referred to as tul4). In contrast to the obligate intracellular genus Wolbachia, strain ATCC VR-331T and facultative intracellular Francisella can replicate in specialized cell-free media. Collectively, these results demonstrate that Wolbachia persica should be reclassified in the genus Francisella as Francisella persica comb. nov. The type strain of Francisella persica comb. nov. is ATCC VR-331T (= FSC845T = DSM 101678T). An emended description of the family Francisellaceae is also provided.}, } @article {pmid26746715, year = {2016}, author = {Wilkinson, DA and Duron, O and Cordonin, C and Gomard, Y and Ramasindrazana, B and Mavingui, P and Goodman, SM and Tortosa, P}, title = {The Bacteriome of Bat Flies (Nycteribiidae) from the Malagasy Region: a Community Shaped by Host Ecology, Bacterial Transmission Mode, and Host-Vector Specificity.}, journal = {Applied and environmental microbiology}, volume = {82}, number = {6}, pages = {1778-1788}, pmid = {26746715}, issn = {1098-5336}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; Chiroptera/parasitology ; Diptera/*microbiology ; Madagascar ; *Microbiota ; }, abstract = {The Nycteribiidae are obligate blood-sucking Diptera (Hippoboscoidea) flies that parasitize bats. Depending on species, these wingless flies exhibit either high specialism or generalism toward their hosts, which may in turn have important consequences in terms of their associated microbial community structure. Bats have been hypothesized to be reservoirs of numerous infectious agents, some of which have recently emerged in human populations. Thus, bat flies may be important in the epidemiology and transmission of some of these bat-borne infectious diseases, acting either directly as arthropod vectors or indirectly by shaping pathogen communities among bat populations. In addition, bat flies commonly have associations with heritable bacterial endosymbionts that inhabit insect cells and depend on maternal transmission through egg cytoplasm to ensure their transmission. Some of these heritable bacteria are likely obligate mutualists required to support bat fly development, but others are facultative symbionts with unknown effects. Here, we present bacterial community profiles that were obtained from seven bat fly species, representing five genera, parasitizing bats from the Malagasy region. The observed bacterial diversity includes Rickettsia, Wolbachia, and several Arsenophonus-like organisms, as well as other members of the Enterobacteriales and a widespread association of Bartonella bacteria from bat flies of all five genera. Using the well-described host specificity of these flies and data on community structure from selected bacterial taxa with either vertical or horizontal transmission, we show that host/vector specificity and transmission mode are important drivers of bacterial community structure.}, } @article {pmid26745630, year = {2016}, author = {Ross, PA and Endersby, NM and Hoffmann, AA}, title = {Costs of Three Wolbachia Infections on the Survival of Aedes aegypti Larvae under Starvation Conditions.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {1}, pages = {e0004320}, pmid = {26745630}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Female ; Food Deprivation ; Host-Pathogen Interactions ; Larva/microbiology ; Male ; Wolbachia/classification/*physiology ; }, abstract = {The mosquito Aedes aegypti, the principal vector of dengue virus, has recently been infected experimentally with Wolbachia: intracellular bacteria that possess potential as dengue biological control agents. Wolbachia depend on their hosts for nutrients they are unable to synthesize themselves. Consequently, competition between Wolbachia and their host for resources could reduce host fitness under the competitive conditions commonly experienced by larvae of Ae. aegypti in the field, hampering the invasion of Wolbachia into natural mosquito populations. We assess the survival and development of Ae. aegypti larvae under starvation conditions when infected with each of three experimentally-generated Wolbachia strains: wMel, wMelPop and wAlbB, and compare their fitness to wild-type uninfected larvae. We find that all three Wolbachia infections reduce the survival of larvae relative to those that are uninfected, and the severity of the effect is concordant with previously characterized fitness costs to other life stages. We also investigate the ability of larvae to recover from extended food deprivation and find no effect of Wolbachia on this trait. Aedes aegypti larvae of all infection types were able to resume their development after one month of no food, pupate rapidly, emerge at a large size, and exhibit complete cytoplasmic incompatibility and maternal transmission. A lowered ability of Wolbachia-infected larvae to survive under starvation conditions will increase the threshold infection frequency required for Wolbachia to establish in highly competitive natural Ae. aegypti populations and will also reduce the speed of invasion. This study also provides insights into survival strategies of larvae when developing in stressful environments.}, } @article {pmid26741364, year = {2016}, author = {Pappalardo, AM and D'Urso, V and Viscuso, R and Ferrito, V and Giunta, MC and Cupani, S and Vitale, DG}, title = {Morphostructural investigation of the female reproductive system and molecular evidence for Wolbachia in Balclutha brevis Lindberg 1954 (Hemiptera, Cicadellidae).}, journal = {Micron (Oxford, England : 1993)}, volume = {81}, number = {}, pages = {23-33}, doi = {10.1016/j.micron.2015.12.003}, pmid = {26741364}, issn = {1878-4291}, mesh = {Animals ; Bacterial Proteins/genetics ; DNA, Bacterial/analysis/genetics ; DNA, Ribosomal/genetics ; Europe ; Female ; Genitalia, Female/anatomy & histology/ultrastructure ; Hemiptera/*microbiology ; Histocytochemistry ; Male ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Molecular Sequence Data ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Balclutha brevis Lindberg 1954 (Homoptera, Cicadellidae) is an allochthonous species that is rapidly spreading in Sicily and in mainland Europe due to the wide spread of its host plant and therefore could also compete with populations of native species. Considering these ecological implications, based on the lacking ultrastructural data about the reproductive systems of the Auchenorrhyncha and since previous investigations on the male reproductive system of B. brevis have shown some interesting features, we carried out morphostructural investigations on the female reproductive system of this alien leafhopper. Moreover, given the high interest in literature on Wolbachia entomoparasite and based on our previous studies, we provided a contribution to further investigations in applied sciences. For this aim we performed a molecular analysis on males and females of B. brevis to detect the possible presence of strains of the bacterium known to alter host reproductive biology. The female reproductive system has a morphological organization comparable to the general anatomical features of most of the Auchenorrhyncha species; however, comparing our data with the literature, some considerations are discussed. As for the histological and ultrastructural investigations, our results show a secretory activity of the various examined structures. In the spermatheca of B. brevis, in particular, the secretory activity is more marked in the sac-shaped tract, where histochemical investigations showed a lipid component of the secretion; possible origin of this component is discussed. Moreover, mainly free spermatozoa are found in the sac-shaped tract of the spermatheca and in the common oviduct. As for the latter, an interesting findings is the lack of cuticular intima on the epithelial surface of the common oviduct; furthermore, the observed features and the literature in this regards led us to review the significance of the structure called as spermatheca. The molecular screening study (PCR) with three Wolbachia specific genes (16SrRNA, ftsZ,wsp) strongly supports Wolbachia infections in both males and females of the host species. This is the first study demonstrating the presence of Wolbachia in Balclutha genus, though no morphostructural alteration commonly related to the presence of the bacterium has been found in all the examined specimens.}, } @article {pmid26733969, year = {2015}, author = {Braquart-Varnier, C and Raimond, M and Mappa, G and Chevalier, FD and Le Clec'h, W and Sicard, M}, title = {The Hematopoietic Organ: A Cornerstone for Wolbachia Propagation Between and Within Hosts.}, journal = {Frontiers in microbiology}, volume = {6}, number = {}, pages = {1424}, pmid = {26733969}, issn = {1664-302X}, abstract = {Wolbachia is an intracellular α-proteobacterium which is transmitted vertically from mother to offspring but also frequently switches horizontally from one host to another. Our hypothesis is based on the role of immune cells and the organs that produce them, the hematopoietic organs (HOs), as primordial niches for the propagation of Wolbachia via hemocytes both (i) within hosts: to initiate and maintain the systemic infection and (ii) between hosts: to promote both vertical and horizontal transmission of Wolbachia. Therefore, we review some fundamental ideas underlying this hypothesis and go further with new empirical data that lead to a first close-up analysis of the potential role of HOs in Wolbachia propagation. The monitoring of the first steps of Wolbachia infection in horizontally infected host organs by transmission electron microscopy and qPCR suggests that (i) HOs are colonized early and extensively as soon as they are in contact with Wolbachia which find in these cells a favorable niche to multiply and (ii) infected HOs which expel hemocytes all lifelong can generate and maintain a systemic infection that could contribute to increase both vertical and horizontal propagation of these symbionts.}, } @article {pmid26733946, year = {2015}, author = {Braquart-Varnier, C and Altinli, M and Pigeault, R and Chevalier, FD and Grève, P and Bouchon, D and Sicard, M}, title = {The Mutualistic Side of Wolbachia-Isopod Interactions: Wolbachia Mediated Protection Against Pathogenic Intracellular Bacteria.}, journal = {Frontiers in microbiology}, volume = {6}, number = {}, pages = {1388}, pmid = {26733946}, issn = {1664-302X}, abstract = {Wolbachia is a vertically transmitted endosymbiont whose radiative success is mainly related to various host reproductive manipulations that led to consider this symbiont as a conflictual reproductive parasite. However, lately, some Wolbachia have been shown to act as beneficial symbionts by protecting hosts against a broad range of parasites. Still, this protection has been mostly demonstrated in artificial Wolbachia-host associations between partners that did not co-evolved together. Here, we tested in two terrestrial isopod species Armadillidium vulgare and Porcellio dilatatus whether resident Wolbachia (native or non-native) could confer protection during infections with Listeria ivanovii and Salmonella typhimurium and also during a transinfection with a Wolbachia strain that kills the recipient host (i.e., wVulC in P. dilatatus). Survival analyses showed that (i) A. vulgare lines hosting their native Wolbachia (wVulC) always exhibited higher survival than asymbiotic ones when infected with pathogenic bacteria (ii) P. dilatatus lines hosting their native wDil Wolbachia strain survived the S. typhimurium infection better, while lines hosting non-native wCon Wolbachia strain survived the L. ivanovii and also the transinfection with wVulC from A. vulgare better. By studying L. ivanovii and S. typhimurium loads in the hemolymph of the different host-Wolbachia systems, we showed that (i) the difference in survival between lines after L. ivanovii infections were not linked to the difference between their pathogenic bacterial loads, and (ii) the difference in survival after S. typhimurium infections corresponds to lower loads of pathogenic bacteria. Overall, our results demonstrate a beneficial effect of Wolbachia on survival of terrestrial isopods when infected with pathogenic intracellular bacteria. This protective effect may rely on different mechanisms depending on the resident symbiont and the invasive bacteria interacting together within the hosts.}, } @article {pmid26721865, year = {2016}, author = {Skelton, E and Rancès, E and Frentiu, FD and Kusmintarsih, ES and Iturbe-Ormaetxe, I and Caragata, EP and Woolfit, M and O'Neill, SL}, title = {A Native Wolbachia Endosymbiont Does Not Limit Dengue Virus Infection in the Mosquito Aedes notoscriptus (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {53}, number = {2}, pages = {401-408}, pmid = {26721865}, issn = {0022-2585}, mesh = {Animals ; Culicidae/*virology ; Dengue Virus/*physiology ; Female ; Ovary/microbiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia pipientis infects many species of insects and has been transinfected into the mosquito Aedes aegypti (L.), the primary vector of dengue virus (DENV). Recently, it has been shown that Wolbachia blocks the replication and transmission of RNA viruses, such as DENV, in a number of mosquito species including Ae. aegypti and Aedes albopictus (Skuse), which is naturally infected with Wolbachia and considered a secondary vector for DENV. The mosquito species Aedes notoscriptus (Skuse) is highly prevalent in Australia, including in areas where DENV outbreaks have been recorded. The mosquito has been implicated in the transmission of Ross River and Barmah Forest viruses, but not DENV. We investigated whether Wolbachia naturally infects this mosquito species and whether it has an impact on the ability of Ae. notoscriptus to transmit DENV. We show, for the first time, that Ae. notoscriptus is naturally infected with a strain of Wolbachia that belongs to supergroup B and is localized only in the ovaries. However, Wolbachia infection in Ae. notoscriptus did not induce resistance to DENV and had no effect on overall DENV infection rate or titer. The presence of a native Wolbachia in Ae. notoscriptus cannot explain why this mosquito is an ineffective vector of DENV.}, } @article {pmid26711515, year = {2016}, author = {Axford, JK and Ross, PA and Yeap, HL and Callahan, AG and Hoffmann, AA}, title = {Fitness of wAlbB Wolbachia Infection in Aedes aegypti: Parameter Estimates in an Outcrossed Background and Potential for Population Invasion.}, journal = {The American journal of tropical medicine and hygiene}, volume = {94}, number = {3}, pages = {507-516}, pmid = {26711515}, issn = {1476-1645}, mesh = {Aedes/*microbiology ; Animals ; Bacterial Load ; Cell Survival ; Female ; Fertility ; *Genetic Fitness ; Host-Pathogen Interactions ; Male ; Ovum/microbiology/physiology ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Wolbachia endosymbionts are potentially useful tools for suppressing disease transmission by Aedes aegypti mosquitoes because Wolbachia can interfere with the transmission of dengue and other viruses as well as causing deleterious effects on their mosquito hosts. Most recent research has focused on the wMel infection, but other infections also influence viral transmission and may spread in natural populations. Here, we focus on the wAlbB infection in an Australian outbred background and show that this infection has many features that facilitate its invasion into natural populations including strong cytoplasmic incompatibility, a lack of effect on larval development, an equivalent mating success to uninfected males and perfect maternal transmission fidelity. On the other hand, the infection has deleterious effects when eggs are held in a dried state, falling between wMel and the more virulent wMelPop Wolbachia strains. The impact of this infection on lifespan also appears to be intermediate, consistent with the observation that this infection has a titer in adults between wMel and wMelPop. Population cage experiments indicate that the wAlbB infection establishes in cages when introduced at a frequency of 22%, suggesting that this strain could be successfully introduced into populations and subsequently persist and spread.}, } @article {pmid26706669, year = {2015}, author = {Tahir, U and Khan, UH and Zubair, MS and Bahar-E-Mustafa, }, title = {Wolbachia pipientis: A potential candidate for combating and eradicating dengue epidemics in Pakistan.}, journal = {Asian Pacific journal of tropical medicine}, volume = {8}, number = {12}, pages = {989-998}, doi = {10.1016/j.apjtm.2015.11.012}, pmid = {26706669}, issn = {2352-4146}, abstract = {Dengue virus syndrome is an emerging global health challenge which is endemic in tropical countries like Pakistan. In recent years dengue incidences have increased considerably in different areas of Pakistan with more sever impacts on urban and peri-urban populations. This review is an effort to highlight the changing epidemiology of dengue fever, role of Government of Pakistan in disease management and control using preventive and community based approaches in the region. Moreover, there is an emphasis on application of Wolbachia as novel, inexpensive and environmentally benign candidate for control and eradication of dengue transmitting vectors.}, } @article {pmid26697448, year = {2015}, author = {Lambrechts, L}, title = {Predicting Wolbachia potential to knock down dengue virus transmission.}, journal = {Annals of translational medicine}, volume = {3}, number = {19}, pages = {288}, pmid = {26697448}, issn = {2305-5839}, abstract = {Releasing mosquitoes infected with the intracellular bacteria Wolbachia is a candidate strategy for dengue control that has recently advanced to field-testing. A critical next step is to evaluate the impact of this strategy on dengue epidemiology. A recent study by Ferguson and colleagues presents a mathematical framework to predict the likely effect of mosquitoes carrying Wolbachia on dengue virus transmission. Fitting the mathematical model to empirical data obtained with Wolbachia-infected mosquitoes experimentally challenged with viremic blood from dengue patients indicates that dengue virus transmission could be reduced by a degree that would have a significant impact on public health.}, } @article {pmid26678796, year = {2016}, author = {Kliot, A and Ghanim, M}, title = {Fluorescent in situ hybridization for the localization of viruses, bacteria and other microorganisms in insect and plant tissues.}, journal = {Methods (San Diego, Calif.)}, volume = {98}, number = {}, pages = {74-81}, doi = {10.1016/j.ymeth.2015.12.003}, pmid = {26678796}, issn = {1095-9130}, mesh = {Animals ; Botrytis/genetics/metabolism/*ultrastructure ; Digoxigenin/chemistry ; Diptera/microbiology/*ultrastructure/virology ; Fluorescent Dyes/chemistry ; Gene Expression Regulation ; In Situ Hybridization, Fluorescence/*methods ; Oligonucleotides/chemistry ; Pisum sativum/microbiology/*ultrastructure/virology ; RNA, Messenger/*chemistry/genetics/metabolism ; Signal-To-Noise Ratio ; Symbiosis ; Tissue Fixation/methods ; Transcription, Genetic ; Weevils/microbiology/*ultrastructure/virology ; Wolbachia/genetics/metabolism/ultrastructure ; }, abstract = {Methods for the localization of cellular components such as nucleic acids, proteins, cellular vesicles and more, and the localization of microorganisms including viruses, bacteria and fungi have become an important part of any research program in biological sciences that enable the visualization of these components in fixed and live tissues without the need for complex processing steps. The rapid development of microscopy tools and technologies as well as related fluorescent markers and fluorophores for many cellular components, and the ability to design DNA and RNA sequence-based molecular probes and antibodies which can be visualized fluorescently, have rapidly advanced this field. This review will focus on some of the localizations methods which have been used in plants and insect pests in agriculture, and other microorganisms, which are rapidly advancing the research in agriculture-related fields.}, } @article {pmid26676774, year = {2015}, author = {Nouri, S and Salem, N and Nigg, JC and Falk, BW}, title = {Diverse Array of New Viral Sequences Identified in Worldwide Populations of the Asian Citrus Psyllid (Diaphorina citri) Using Viral Metagenomics.}, journal = {Journal of virology}, volume = {90}, number = {5}, pages = {2434-2445}, pmid = {26676774}, issn = {1098-5514}, mesh = {Animals ; *Biodiversity ; Cluster Analysis ; Computational Biology ; Gene Expression Profiling ; Hemiptera/*virology ; Insect Viruses/*classification/genetics/*isolation & purification ; Metagenomics/*methods ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, RNA ; }, abstract = {UNLABELLED: The Asian citrus psyllid, Diaphorina citri, is the natural vector of the causal agent of Huanglongbing (HLB), or citrus greening disease. Together; HLB and D. citri represent a major threat to world citrus production. As there is no cure for HLB, insect vector management is considered one strategy to help control the disease, and D. citri viruses might be useful. In this study, we used a metagenomic approach to analyze viral sequences associated with the global population of D. citri. By sequencing small RNAs and the transcriptome coupled with bioinformatics analysis, we showed that the virus-like sequences of D. citri are diverse. We identified novel viral sequences belonging to the picornavirus superfamily, the Reoviridae, Parvoviridae, and Bunyaviridae families, and an unclassified positive-sense single-stranded RNA virus. Moreover, a Wolbachia prophage-related sequence was identified. This is the first comprehensive survey to assess the viral community from worldwide populations of an agricultural insect pest. Our results provide valuable information on new putative viruses, some of which may have the potential to be used as biocontrol agents.

IMPORTANCE: Insects have the most species of all animals, and are hosts to, and vectors of, a great variety of known and unknown viruses. Some of these most likely have the potential to be important fundamental and/or practical resources. In this study, we used high-throughput next-generation sequencing (NGS) technology and bioinformatics analysis to identify putative viruses associated with Diaphorina citri, the Asian citrus psyllid. D. citri is the vector of the bacterium causing Huanglongbing (HLB), currently the most serious threat to citrus worldwide. Here, we report several novel viral sequences associated with D. citri.}, } @article {pmid26675213, year = {2015}, author = {Templé, N and Richard, FJ}, title = {Intra-cellular bacterial infections affect learning and memory capacities of an invertebrate.}, journal = {Frontiers in zoology}, volume = {12}, number = {}, pages = {36}, pmid = {26675213}, issn = {1742-9994}, abstract = {BACKGROUND: How host manipulation by parasites evolves is fascinating but challenging evolutionary question remains. Many parasites share the capacity to manipulate host behavior increasing their transmission success. However, little is known about the learning and memory impact of parasites on their host. Wolbachia are widespread endosymbionts and infect most insect species. These bacteria are maternally transmitted and mainly alter the reproduction of hosts with weak virulence. We tested the impact of parasites (Wolbachia) on their host learning and memory capacities. To address this question we trained individuals to one direction with positive reinforcement. We compared performances between individual Wolbachia-free, Wolbachia naturally and Wolbachia artificially infected individuals.

RESULTS: We report that in the host parasite interaction (Armadillidium vulgare/Wolbachia) naturally infected individuals Wolbachia or transinfected adult with Wolbachia are less likely to learn and memorize the correct direction with social reinforcement compared to Wolbachia-free individuals.

CONCLUSIONS: Our results imply that Wolbachia impact in the central nervous system of their host altering the memory formation and maintenance. We conclude that host manipulation can affect cognitive processes decreasing host adaptation capacities.}, } @article {pmid26670313, year = {2016}, author = {Raharimalala, FN and Boukraa, S and Bawin, T and Boyer, S and Francis, F}, title = {Molecular detection of six (endo-) symbiotic bacteria in Belgian mosquitoes: first step towards the selection of appropriate paratransgenesis candidates.}, journal = {Parasitology research}, volume = {115}, number = {4}, pages = {1391-1399}, pmid = {26670313}, issn = {1432-1955}, mesh = {Animals ; Bacteria/classification/*isolation & purification ; Belgium ; Culicidae/*microbiology ; Insect Control ; Larva/microbiology ; Pest Control, Biological/methods ; Symbiosis ; }, abstract = {Actually, the use of symbiotic bacteria is one of alternative solution to avoid vector resistance to pesticides. In Belgium, among 31 identified mosquito species, 10 were considered as potential vectors. Given to introduction risks of arbovirosis, the purpose of this study was to investigate the presence of symbiosis bacteria in potential mosquito vectors. Eleven species caught from 12 sites in Belgium were used: Culex pipiens s.l., Culex torrentium, Culex hortensis, Anopheles claviger, Anopheles maculipennis s.l., Anopheles plumbeus, Culiseta annulata, Ochlerotatus geniculatus, Ochlerotatus dorsalis, Aedes albopictus, and Coquillettidia richiardii. Six genera of symbiotic bacteria were screened: Wolbachia sp., Comamonas sp, Delftia sp., Pseudomonas sp., Acinetobacter sp., and Asaia sp. A total of 173 mosquito individuals (144 larvae and 29 adults) were used for the polymerase chain reaction screening. Wolbachia was not found in any Anopheles species nor Cx. torrentium. A total absence of Comamonas and Delftia was observed in all species. Acinetobacter, Pseudomonas, and Asaia were found in most of species with a high prevalence for Pseudomonas. These results were discussed to develop potential strategy and exploit the variable occurrence of symbiotic bacteria to focus on them to propose biological ways of mosquito control.}, } @article {pmid26667400, year = {2015}, author = {De Clerck, C and Fujiwara, A and Joncour, P and Léonard, S and Félix, ML and Francis, F and Jijakli, MH and Tsuchida, T and Massart, S}, title = {A metagenomic approach from aphid's hemolymph sheds light on the potential roles of co-existing endosymbionts.}, journal = {Microbiome}, volume = {3}, number = {}, pages = {63}, pmid = {26667400}, issn = {2049-2618}, mesh = {Animals ; Aphids/*microbiology/*physiology ; Babuvirus ; Buchnera/genetics/*physiology ; Genes, Bacterial ; Hemolymph/*microbiology/physiology ; High-Throughput Nucleotide Sequencing ; Metagenome ; *Metagenomics ; Musa ; Riboflavin/metabolism ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Aphids are known to live in symbiosis with specific bacteria, called endosymbionts which can be classified as obligate or accessory. Buchnera aphidicola is generally the only obligatory symbiont present in aphids, supplying essential nutrients that are missing in the plants phloem to its host. Pentalonia nigronervosa is the main vector of the banana bunchy top virus, one of the most damageable viruses in banana. This aphid is carrying two symbionts: B. aphidicola (BPn) and Wolbachia sp. (wPn). The high occurrence of Wolbachia in the banana aphid raises questions about the role it plays in this insect. The goal of this study was to go further in the understanding of the role played by the two symbionts in P. nigronervosa. To do so, microinjection tests were made to see the effect of wPn elimination on the host, and then, high-throughput sequencing of the haemolymph was used to analyze the gene content of the symbionts.

RESULTS: We observed that the elimination of wPn systematically led to the death of aphids, suggesting that the bacterium could play a mutualistic role. In addition, we identify and annotate 587 and 250 genes for wPn and BPn, respectively, through high-throughput sequencing. Analysis of these genes suggests that the two bacteria are working together for the production of several essential nutrients. The most striking cases are for lysin and riboflavin which are usually provided by B. aphidicola alone to the host. In the banana aphid, the genes involved in the production pathways of these metabolites are shared between the two bacteria making them both essential for the survival of the aphid host.

CONCLUSIONS: Our results suggest that a co-obligatory symbiosis between B. aphidicola and Wolbachia occurs in the banana aphid, the two bacteria acting together to supply essential nutrients to the host. This is, to our knowledge, the first time Wolbachia is reported to play an essential role in aphids.}, } @article {pmid26664808, year = {2015}, author = {Funkhouser-Jones, LJ and Sehnert, SR and Martínez-Rodríguez, P and Toribio-Fernández, R and Pita, M and Bella, JL and Bordenstein, SR}, title = {Wolbachia co-infection in a hybrid zone: discovery of horizontal gene transfers from two Wolbachia supergroups into an animal genome.}, journal = {PeerJ}, volume = {3}, number = {}, pages = {e1479}, pmid = {26664808}, issn = {2167-8359}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; T32 GM008554/GM/NIGMS NIH HHS/United States ; }, abstract = {Hybrid zones and the consequences of hybridization have contributed greatly to our understanding of evolutionary processes. Hybrid zones also provide valuable insight into the dynamics of symbiosis since each subspecies or species brings its unique microbial symbionts, including germline bacteria such as Wolbachia, to the hybrid zone. Here, we investigate a natural hybrid zone of two subspecies of the meadow grasshopper Chorthippus parallelus in the Pyrenees Mountains. We set out to test whether co-infections of B and F Wolbachia in hybrid grasshoppers enabled horizontal transfer of phage WO, similar to the numerous examples of phage WO transfer between A and B Wolbachia co-infections. While we found no evidence for transfer between the divergent co-infections, we discovered horizontal transfer of at least three phage WO haplotypes to the grasshopper genome. Subsequent genome sequencing of uninfected grasshoppers uncovered the first evidence for two discrete Wolbachia supergroups (B and F) contributing at least 448 kb and 144 kb of DNA, respectively, into the host nuclear genome. Fluorescent in situ hybridization verified the presence of Wolbachia DNA in C. parallelus chromosomes and revealed that some inserts are subspecies-specific while others are present in both subspecies. We discuss our findings in light of symbiont dynamics in an animal hybrid zone.}, } @article {pmid26664683, year = {2015}, author = {Sontowski, R and Bernhard, D and Bleidorn, C and Schlegel, M and Gerth, M}, title = {Wolbachia distribution in selected beetle taxa characterized by PCR screens and MLST data.}, journal = {Ecology and evolution}, volume = {5}, number = {19}, pages = {4345-4353}, pmid = {26664683}, issn = {2045-7758}, abstract = {Wolbachia (Alphaproteobacteria) is an inherited endosymbiont of arthropods and filarial nematodes and was reported to be widespread across insect taxa. While Wolbachia's effects on host biology are not understood from most of these hosts, known Wolbachia-induced phenotypes cover a spectrum from obligate beneficial mutualism to reproductive manipulations and pathogenicity. Interestingly, data on Wolbachia within the most species-rich order of arthropods, the Coleoptera (beetles), are scarce. Therefore, we screened 128 species from seven beetle families (Buprestidae, Hydraenidae, Dytiscidae, Hydrophilidae, Gyrinidae, Haliplidae, and Noteridae) for the presence of Wolbachia. Our data show that, contrary to previous estimations, Wolbachia frequencies in beetles (31% overall) are comparable to the ones in other insects. In addition, we used Wolbachia MLST data and host phylogeny to explore the evolutionary history of Wolbachia strains from Hydraenidae, an aquatic lineage of beetles. Our data suggest that Wolbachia from Hydraenidae might be largely host genus specific and that Wolbachia strain phylogeny is not independent to that of its hosts. As this contrasts with most terrestrial Wolbachia-arthropod systems, one potential conclusion is that aquatic lifestyle of hosts may result in Wolbachia distribution patterns distinct from those of terrestrial hosts. Our data thus provide both insights into Wolbachia distribution among beetles in general and a first glimpse of Wolbachia distribution patterns among aquatic host lineages.}, } @article {pmid26631376, year = {2015}, author = {Comandatore, F and Cordaux, R and Bandi, C and Blaxter, M and Darby, A and Makepeace, BL and Montagna, M and Sassera, D}, title = {Supergroup C Wolbachia, mutualist symbionts of filarial nematodes, have a distinct genome structure.}, journal = {Open biology}, volume = {5}, number = {12}, pages = {150099}, pmid = {26631376}, issn = {2046-2441}, support = {095831//Wellcome Trust/United Kingdom ; G0900740/MRC_/Medical Research Council/United Kingdom ; MR/K001744/1/MRC_/Medical Research Council/United Kingdom ; //Cancer Research UK/United Kingdom ; }, mesh = {Animals ; Bacterial Proteins/genetics ; Filarioidea/*microbiology ; Genomics ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {Wolbachia pipientis is possibly the most widespread endosymbiont of arthropods and nematodes. While all Wolbachia strains have historically been defined as a single species, 16 monophyletic clusters of diversity (called supergroups) have been described. Different supergroups have distinct host ranges and symbiotic relationships, ranging from mutualism to reproductive manipulation. In filarial nematodes, which include parasites responsible for major diseases of humans (such as Onchocerca volvulus, agent of river blindness) and companion animals (Dirofilaria immitis, the dog heartworm), Wolbachia has an obligate mutualist role and is the target of new treatment regimens. Here, we compare the genomes of eight Wolbachia strains, spanning the diversity of the major supergroups (A-F), analysing synteny, transposable element content, GC skew and gene loss or gain. We detected genomic features that differ between Wolbachia supergroups, most notably in the C and D clades from filarial nematodes. In particular, strains from supergroup C (symbionts of O. volvulus and D. immitis) present a pattern of GC skew, conserved synteny and lack of transposable elements, unique in the Wolbachia genus. These features could be the consequence of a distinct symbiotic relationship between C Wolbachia strains and their hosts, highlighting underappreciated differences between the mutualistic supergroups found within filarial nematodes.}, } @article {pmid26627473, year = {2015}, author = {Rašić, G and Endersby-Harshman, N and Tantowijoyo, W and Goundar, A and White, V and Yang, Q and Filipović, I and Johnson, P and Hoffmann, AA and Arguni, E}, title = {Aedes aegypti has spatially structured and seasonally stable populations in Yogyakarta, Indonesia.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {610}, pmid = {26627473}, issn = {1756-3305}, mesh = {Aedes/classification/genetics/*growth & development ; Animals ; *Genetic Variation ; Genetics, Population ; Indonesia ; *Insect Vectors ; Microsatellite Repeats ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; *Population Dynamics ; *Seasons ; Sequence Analysis, DNA ; Spatio-Temporal Analysis ; Wolbachia ; }, abstract = {BACKGROUND: Dengue fever, the most prevalent global arboviral disease, represents an important public health problem in Indonesia. Control of dengue relies on the control of its main vector, the mosquito Aedes aegypti, yet nothing is known about the population history and genetic structure of this insect in Indonesia. Our aim was to assess the spatio-temporal population genetic structure of Ae. aegypti in Yogyakarta, a densely populated region on Java with common dengue outbreaks.

METHODS: We used multiple marker systems (microsatellites, nuclear and mitochondrial genome-wide single nucleotide polymorphisms generated via Restriction-site Associated DNA sequencing) to analyze 979 Ae. aegypti individuals collected from the Yogyakarta city and the surrounding hamlets during the wet season in 2011 and the following dry season in 2012. We employed individual- and group-based approaches for inferring genetic structure.

RESULTS: We found that Ae. aegypti in Yogyakarta has spatially structured and seasonally stable populations. The spatial structuring was significant for the nuclear and mitochondrial markers, while the temporal structuring was non-significant. Nuclear markers identified three main genetic clusters, showing that hamlets have greater genetic isolation from each other and from the inner city sites. However, one hamlet experienced unrestricted mosquito interbreeding with the inner city, forming a single genetic cluster. Genetic distance was poorly correlated with the spatial distance among mosquito samples, suggesting stronger influence of human-assisted gene flow than active mosquito movement on spatial genetic structure. A star-shaped mitochondrial haplotype network and a significant R(2) test statistic (R(2) = 0.0187, P = 0.001) support the hypothesis that Ae. aegypti in Yogyakarta originated from a small or homogeneous source and has undergone a relatively recent demographic expansion.

CONCLUSION: We report the first insights into the spatio-temporal genetic structure and the underlying processes in the dengue fever mosquito from Yogyakarta, Indonesia. Our results provide valuable information on the effectiveness of local control measures as well as guidelines for the implementation of novel biocontrol strategies such as release of Wolbachia-infected mosquitoes.}, } @article {pmid26627243, year = {2015}, author = {Mesquita, RD and Vionette-Amaral, RJ and Lowenberger, C and Rivera-Pomar, R and Monteiro, FA and Minx, P and Spieth, J and Carvalho, AB and Panzera, F and Lawson, D and Torres, AQ and Ribeiro, JM and Sorgine, MH and Waterhouse, RM and Montague, MJ and Abad-Franch, F and Alves-Bezerra, M and Amaral, LR and Araujo, HM and Araujo, RN and Aravind, L and Atella, GC and Azambuja, P and Berni, M and Bittencourt-Cunha, PR and Braz, GR and Calderón-Fernández, G and Carareto, CM and Christensen, MB and Costa, IR and Costa, SG and Dansa, M and Daumas-Filho, CR and De-Paula, IF and Dias, FA and Dimopoulos, G and Emrich, SJ and Esponda-Behrens, N and Fampa, P and Fernandez-Medina, RD and da Fonseca, RN and Fontenele, M and Fronick, C and Fulton, LA and Gandara, AC and Garcia, ES and Genta, FA and Giraldo-Calderón, GI and Gomes, B and Gondim, KC and Granzotto, A and Guarneri, AA and Guigó, R and Harry, M and Hughes, DS and Jablonka, W and Jacquin-Joly, E and Juárez, MP and Koerich, LB and Lange, AB and Latorre-Estivalis, JM and Lavore, A and Lawrence, GG and Lazoski, C and Lazzari, CR and Lopes, RR and Lorenzo, MG and Lugon, MD and Majerowicz, D and Marcet, PL and Mariotti, M and Masuda, H and Megy, K and Melo, AC and Missirlis, F and Mota, T and Noriega, FG and Nouzova, M and Nunes, RD and Oliveira, RL and Oliveira-Silveira, G and Ons, S and Orchard, I and Pagola, L and Paiva-Silva, GO and Pascual, A and Pavan, MG and Pedrini, N and Peixoto, AA and Pereira, MH and Pike, A and Polycarpo, C and Prosdocimi, F and Ribeiro-Rodrigues, R and Robertson, HM and Salerno, AP and Salmon, D and Santesmasses, D and Schama, R and Seabra-Junior, ES and Silva-Cardoso, L and Silva-Neto, MA and Souza-Gomes, M and Sterkel, M and Taracena, ML and Tojo, M and Tu, ZJ and Tubio, JM and Ursic-Bedoya, R and Venancio, TM and Walter-Nuno, AB and Wilson, D and Warren, WC and Wilson, RK and Huebner, E and Dotson, EM and Oliveira, PL}, title = {Genome of Rhodnius prolixus, an insect vector of Chagas disease, reveals unique adaptations to hematophagy and parasite infection.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {112}, number = {48}, pages = {14936-14941}, pmid = {26627243}, issn = {1091-6490}, support = {NHGRI-HG003079//PHS HHS/United States ; HHSN272200900039C//PHS HHS/United States ; R01 AI045545/AI/NIAID NIH HHS/United States ; HHSN272200900039C/AI/NIAID NIH HHS/United States ; U54 HG003079/HG/NHGRI NIH HHS/United States ; }, mesh = {Adaptation, Physiological/*genetics ; Animals ; Base Sequence ; *Chagas Disease ; Gene Transfer, Horizontal ; Host-Parasite Interactions/*genetics ; Humans ; *Insect Vectors/genetics/parasitology ; Molecular Sequence Data ; *Rhodnius/genetics/parasitology ; Trypanosoma cruzi/*physiology ; Wolbachia/genetics ; }, abstract = {Rhodnius prolixus not only has served as a model organism for the study of insect physiology, but also is a major vector of Chagas disease, an illness that affects approximately seven million people worldwide. We sequenced the genome of R. prolixus, generated assembled sequences covering 95% of the genome (∼ 702 Mb), including 15,456 putative protein-coding genes, and completed comprehensive genomic analyses of this obligate blood-feeding insect. Although immune-deficiency (IMD)-mediated immune responses were observed, R. prolixus putatively lacks key components of the IMD pathway, suggesting a reorganization of the canonical immune signaling network. Although both Toll and IMD effectors controlled intestinal microbiota, neither affected Trypanosoma cruzi, the causal agent of Chagas disease, implying the existence of evasion or tolerance mechanisms. R. prolixus has experienced an extensive loss of selenoprotein genes, with its repertoire reduced to only two proteins, one of which is a selenocysteine-based glutathione peroxidase, the first found in insects. The genome contained actively transcribed, horizontally transferred genes from Wolbachia sp., which showed evidence of codon use evolution toward the insect use pattern. Comparative protein analyses revealed many lineage-specific expansions and putative gene absences in R. prolixus, including tandem expansions of genes related to chemoreception, feeding, and digestion that possibly contributed to the evolution of a blood-feeding lifestyle. The genome assembly and these associated analyses provide critical information on the physiology and evolution of this important vector species and should be instrumental for the development of innovative disease control methods.}, } @article {pmid26602337, year = {2015}, author = {Almeida, RP and Stouthamer, R}, title = {ITS-2 sequences-based identification of Trichogramma species in South America.}, journal = {Brazilian journal of biology = Revista brasleira de biologia}, volume = {75}, number = {4}, pages = {974-982}, doi = {10.1590/1519-6984.04614}, pmid = {26602337}, issn = {1678-4375}, mesh = {Animals ; DNA, Ribosomal Spacer/genetics ; Molecular Sequence Data ; Parthenogenesis ; Sequence Analysis, DNA ; South America ; Wasps/*classification/genetics/microbiology/*physiology ; Wolbachia/physiology ; }, abstract = {ITS2 (Internal transcribed spacer 2) sequences have been used in systematic studies and proved to be useful in providing a reliable identification of Trichogramma species. DNAr sequences ranged in size from 379 to 632 bp. In eleven T. pretiosum lines Wolbachia-induced parthenogenesis was found for the first time. These thelytokous lines were collected in Peru (9), Colombia (1) and USA (1). A dichotomous key for species identification was built based on the size of the ITS2 PCR product and restriction analysis using three endonucleases (EcoRI, MseI and MaeI). This molecular technique was successfully used to distinguish among seventeen native/introduced Trichogramma species collected in South America.}, } @article {pmid26601484, year = {2015}, author = {Goryacheva, II and Blekhman, AV and Andrianov, BV and Gorelova, TV and Zakharov, IA}, title = {[Genotypic Diversity of Wolbachia pipientis in Native and Invasive Harmonia axyridis Pall., 1773 (Coleoptera, Coccinellidae) Populations].}, journal = {Genetika}, volume = {51}, number = {8}, pages = {857-863}, pmid = {26601484}, issn = {0016-6758}, mesh = {Animals ; Bacterial Proteins/genetics ; Coleoptera/*genetics/microbiology ; *Evolution, Molecular ; Genetic Variation ; Genetics, Population ; Genotype ; Phylogeny ; Reproduction/genetics ; Symbiosis/*genetics ; Wolbachia/*genetics ; }, abstract = {The distribution and variability of reproductive symbiotic Wolbachia pipientis bacteria were studied in seven native and six invasive H. axyridis populations. Wolbachia-infected individuals were found in two invasive and two native populations. We demonstrated for the first time an infection of invasive H. axyridis populations with Wolbachia. Two new molecular forms of Wolbachia were detected by a system of multilocus typing. The supergroup A Wolbachia was found for the first time in H. axyridis. The detected genotypic diversity of Wolbachia indicates repeated and independent infection events in the evolutionary past of H. axyridis.}, } @article {pmid26579107, year = {2015}, author = {Zug, R and Hammerstein, P}, title = {Wolbachia and the insect immune system: what reactive oxygen species can tell us about the mechanisms of Wolbachia-host interactions.}, journal = {Frontiers in microbiology}, volume = {6}, number = {}, pages = {1201}, pmid = {26579107}, issn = {1664-302X}, abstract = {Wolbachia are intracellular bacteria that infect a vast range of arthropod species, making them one of the most prevalent endosymbionts in the world. Wolbachia's stunning evolutionary success is mostly due to their reproductive parasitism but also to mutualistic effects such as increased host fecundity or protection against pathogens. However, the mechanisms underlying Wolbachia phenotypes, both parasitic and mutualistic, are only poorly understood. Moreover, it is unclear how the insect immune system is involved in these phenotypes and why it is not more successful in eliminating the bacteria. Here we argue that reactive oxygen species (ROS) are likely to be key in elucidating these issues. ROS are essential players in the insect immune system, and Wolbachia infection can affect ROS levels in the host. Based on recent findings, we elaborate a hypothesis that considers the different effects of Wolbachia on the oxidative environment in novel vs. native hosts. We propose that newly introduced Wolbachia trigger an immune response and cause oxidative stress, whereas in coevolved symbioses, infection is not associated with oxidative stress, but rather with restored redox homeostasis. Redox homeostasis can be restored in different ways, depending on whether Wolbachia or the host is in charge. This hypothesis offers a mechanistic explanation for several of the observed Wolbachia phenotypes.}, } @article {pmid26576934, year = {2016}, author = {Tantowijoyo, W and Arguni, E and Johnson, P and Budiwati, N and Nurhayati, PI and Fitriana, I and Wardana, S and Ardiansyah, H and Turley, AP and Ryan, P and O'Neill, SL and Hoffmann, AA}, title = {Spatial and Temporal Variation in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) Numbers in the Yogyakarta Area of Java, Indonesia, With Implications for Wolbachia Releases.}, journal = {Journal of medical entomology}, volume = {53}, number = {1}, pages = {188-198}, doi = {10.1093/jme/tjv180}, pmid = {26576934}, issn = {0022-2585}, mesh = {*Aedes ; Animals ; Indonesia ; Pest Control, Biological ; Population Density ; Population Dynamics ; Wolbachia ; }, abstract = {of mosquito vector populations, particularly through Wolbachia endosymbionts. The success of these strategies depends on understanding the dynamics of vector populations. In preparation for Wolbachia releases around Yogyakarta, we have studied Aedes populations in five hamlets. Adult monitoring with BioGent- Sentinel (BG-S) traps indicated that hamlet populations had different dynamics across the year; while there was an increase in Aedes aegypti (L.) and Aedes albopictus (Skuse) numbers in the wet season, species abundance remained relatively stable in some hamlets but changed markedly (>2 fold) in others. Local rainfall a month prior to monitoring partly predicted numbers of Ae. aegypti but not Ae. albopictus. Site differences in population size indicated by BG-S traps were also evident in ovitrap data. Egg or larval collections with ovitraps repeated at the same location suggested spatial autocorrelation (<250 m) in the areas of the hamlets where Ae. aegypti numbers were high. Overall, there was a weak negative association (r<0.43) between Ae. aegypti and Ae. albopictus numbers in ovitraps when averaged across collections. Ae. albopictus numbers in ovitraps and BG-S traps were positively correlated with vegetation around areas where traps were placed, while Ae. aegypti were negatively correlated with this feature. These data inform intervention strategies by defining periods when mosquito densities are high, highlighting the importance of local site characteristics on populations, and suggesting relatively weak interactions between Ae. aegypti and Ae. albopictus. They also indicate local areas within hamlets where consistently high mosquito densities may influence Wolbachia invasions and other interventions.}, } @article {pmid26573516, year = {2016}, author = {Qiu, Y and Nakao, R and Thu, MJ and Akter, S and Alam, MZ and Kato, S and Katakura, K and Sugimoto, C}, title = {Molecular evidence of spotted fever group rickettsiae and Anaplasmataceae from ticks and stray dogs in Bangladesh.}, journal = {Parasitology research}, volume = {115}, number = {3}, pages = {949-955}, pmid = {26573516}, issn = {1432-1955}, mesh = {Anaplasmataceae/classification/genetics/*isolation & purification ; Anaplasmataceae Infections/microbiology/transmission/*veterinary ; Animals ; Arachnid Vectors/*microbiology ; Bangladesh ; Base Sequence ; Cattle ; Dog Diseases/*microbiology/transmission ; Dogs ; Ixodidae/*microbiology ; Real-Time Polymerase Chain Reaction ; Rickettsia/classification/genetics/*isolation & purification ; Rickettsia Infections/microbiology/transmission/*veterinary ; Tick-Borne Diseases/microbiology/transmission/*veterinary ; }, abstract = {Emerging tick-borne diseases (TBDs) are important foci for human and animal health worldwide. However, these diseases are sometimes over looked, especially in countries with limited resources to perform molecular-based surveys. The aim of this study was to detect and characterize spotted fever group (SFG) rickettsiae and Anaplasmataceae in Bangladesh, which are important tick-borne pathogens for humans and animals worldwide. A total of 50 canine blood samples, 15 ticks collected from dogs, and 154 ticks collected from cattle were screened for the presence of SFG rickettsiae and Anaplasmataceae using molecular-based methods such as PCR and real-time PCR. The sequence analysis of the amplified products detected two different genotypes of SFG rickettsiae in ticks from cattle. The genotype detected in Rhipicephalus microplus was closely related to Rickettsia monacensis, while the genotype detected in Haemaphysalis bispinosa was closely related to Rickettsia sp. found in Korea and Japan. Anaplasma bovis was detected in canine blood and ticks (Rhipicephalus sanguineus and H. bispinosa). Unexpectedly, the partial genome sequence of Wolbachia sp., presumably associated with the nematode Dirofilaria immitis, was identified in canine blood. The present study provides the first molecular evidence of SFG rickettsiae and A. bovis in Bangladesh, indicating the possible emergence of previously unrecognized TBDs in this country.}, } @article {pmid26563507, year = {2015}, author = {Gonella, E and Pajoro, M and Marzorati, M and Crotti, E and Mandrioli, M and Pontini, M and Bulgari, D and Negri, I and Sacchi, L and Chouaia, B and Daffonchio, D and Alma, A}, title = {Plant-mediated interspecific horizontal transmission of an intracellular symbiont in insects.}, journal = {Scientific reports}, volume = {5}, number = {}, pages = {15811}, pmid = {26563507}, issn = {2045-2322}, mesh = {Animals ; Bacteroidetes/classification/genetics/*physiology ; Geography ; Hemiptera/genetics/*microbiology/*physiology ; Host-Pathogen Interactions ; In Situ Hybridization ; In Situ Hybridization, Fluorescence ; Insect Vectors/microbiology/physiology ; Intracellular Space/microbiology/parasitology ; Italy ; Microscopy, Electron, Transmission ; Phylogeny ; Plant Diseases/microbiology/parasitology ; Plant Leaves/microbiology/parasitology ; Plants/*microbiology/*parasitology ; RNA, Ribosomal, 16S/genetics ; Real-Time Polymerase Chain Reaction ; Salivary Glands/microbiology ; Symbiosis ; Vitis/microbiology/parasitology ; }, abstract = {Intracellular reproductive manipulators, such as Candidatus Cardinium and Wolbachia are vertically transmitted to progeny but rarely show co-speciation with the host. In sap-feeding insects, plant tissues have been proposed as alternative horizontal routes of interspecific transmission, but experimental evidence is limited. Here we report results from experiments that show that Cardinium is horizontally transmitted between different phloem sap-feeding insect species through plants. Quantitative PCR and in situ hybridization experiments indicated that the leafhopper Scaphoideus titanus releases Cardinium from its salivary glands during feeding on both artificial media and grapevine leaves. Successional time-course feeding experiments with S. titanus initially fed sugar solutions or small areas of grapevine leaves followed by feeding by the phytoplasma vector Macrosteles quadripunctulatus or the grapevine feeder Empoasca vitis revealed that the symbionts were transmitted to both species. Explaining interspecific horizontal transmission through plants improves our understanding of how symbionts spread, their lifestyle and the symbiont-host intermixed evolutionary pattern.}, } @article {pmid26559510, year = {2015}, author = {Luck, AN and Anderson, KG and McClung, CM and VerBerkmoes, NC and Foster, JM and Michalski, ML and Slatko, BE}, title = {Tissue-specific transcriptomics and proteomics of a filarial nematode and its Wolbachia endosymbiont.}, journal = {BMC genomics}, volume = {16}, number = {}, pages = {920}, pmid = {26559510}, issn = {1471-2164}, mesh = {Animals ; Cluster Analysis ; Computational Biology/methods ; Dirofilaria immitis/*genetics/*metabolism ; Female ; Gene Expression Profiling ; Male ; Organ Specificity/genetics ; *Proteome ; Proteomics ; *Symbiosis ; *Transcriptome ; Wolbachia/*genetics/*metabolism ; }, abstract = {BACKGROUND: Filarial nematodes cause debilitating human diseases. While treatable, recent evidence suggests drug resistance is developing, necessitating the development of novel targets and new treatment options. Although transcriptomic and proteomic studies around the nematode life cycle have greatly enhanced our knowledge, whole organism approaches have not provided spatial resolution of gene expression, which can be gained by examining individual tissues. Generally, due to their small size, tissue dissection of human-infecting filarial nematodes remains extremely challenging. However, canine heartworm disease is caused by a closely related and much larger filarial nematode, Dirofilaria immitis. As with many other filarial nematodes, D. immitis contains Wolbachia, an obligate bacterial endosymbiont present in the hypodermis and developing oocytes within the uterus. Here, we describe the first concurrent tissue-specific transcriptomic and proteomic profiling of a filarial nematode (D. immitis) and its Wolbachia (wDi) in order to better understand tissue functions and identify tissue-specific antigens that may be used for the development of new diagnostic and therapeutic tools.

METHODS: Adult D. immitis worms were dissected into female body wall (FBW), female uterus (FU), female intestine (FI), female head (FH), male body wall (MBW), male testis (MT), male intestine (MI), male head (MH) and 10.1186/s12864-015-2083-2 male spicule (MS) and used to prepare transcriptomic and proteomic libraries.

RESULTS: Transcriptomic and proteomic analysis of several D. immitis tissues identified many biological functions enriched within certain tissues. Hierarchical clustering of the D. immitis tissue transcriptomes, along with the recently published whole-worm adult male and female D. immitis transcriptomes, revealed that the whole-worm transcriptome is typically dominated by transcripts originating from reproductive tissue. The uterus appeared to have the most variable transcriptome, possibly due to age. Although many functions are shared between the reproductive tissues, the most significant differences in gene expression were observed between the uterus and testis. Interestingly, wDi gene expression in the male and female body wall is fairly similar, yet slightly different to that of Wolbachia gene expression in the uterus. Proteomic methods verified 32 % of the predicted D. immitis proteome, including over 700 hypothetical proteins of D. immitis. Of note, hypothetical proteins were among some of the most abundant Wolbachia proteins identified, which may fulfill some important yet still uncharacterized biological function.

CONCLUSIONS: The spatial resolution gained from this parallel transcriptomic and proteomic analysis adds to our understanding of filarial biology and serves as a resource with which to develop future therapeutic strategies against filarial nematodes and their Wolbachia endosymbionts.}, } @article {pmid26556361, year = {2015}, author = {Johnson, KN}, title = {The Impact of Wolbachia on Virus Infection in Mosquitoes.}, journal = {Viruses}, volume = {7}, number = {11}, pages = {5705-5717}, pmid = {26556361}, issn = {1999-4915}, mesh = {Animals ; Culicidae/*microbiology/*virology ; Insect Vectors/*microbiology/*virology ; *Symbiosis ; Viruses/*growth & development ; Wolbachia/growth & development/*physiology ; }, abstract = {Mosquito-borne viruses such as dengue, West Nile and chikungunya viruses cause significant morbidity and mortality in human populations. Since current methods are not sufficient to control disease occurrence, novel methods to control transmission of arboviruses would be beneficial. Recent studies have shown that virus infection and transmission in insects can be impeded by co-infection with the bacterium Wolbachia pipientis. Wolbachia is a maternally inherited endosymbiont that is commonly found in insects, including a number of mosquito vector species. In Drosophila, Wolbachia mediates antiviral protection against a broad range of RNA viruses. This discovery pointed to a potential strategy to interfere with mosquito transmission of arboviruses by artificially infecting mosquitoes with Wolbachia. This review outlines research on the prevalence of Wolbachia in mosquito vector species and the impact of antiviral effects in both naturally and artificially Wolbachia-infected mosquitoes.}, } @article {pmid26556278, year = {2015}, author = {Moriyama, M and Nikoh, N and Hosokawa, T and Fukatsu, T}, title = {Riboflavin Provisioning Underlies Wolbachia's Fitness Contribution to Its Insect Host.}, journal = {mBio}, volume = {6}, number = {6}, pages = {e01732-15}, pmid = {26556278}, issn = {2150-7511}, mesh = {Animals ; Biosynthetic Pathways/genetics ; Evolution, Molecular ; Insecta/*microbiology/*physiology ; Riboflavin/*metabolism ; *Symbiosis ; Wolbachia/genetics/*metabolism/*physiology ; }, abstract = {UNLABELLED: Endosymbiotic bacteria of the genus Wolbachia represent the most successful symbiotic bacteria in the terrestrial ecosystem. The success of Wolbachia has been ascribed to its remarkable phenotypic effects on host reproduction, such as cytoplasmic incompatibility, whereby maternally inherited bacteria can spread in their host populations at the expense of their host's fitness. Meanwhile, recent theoretical as well as empirical studies have unveiled that weak and/or conditional positive fitness effects may significantly facilitate invasion and spread of Wolbachia infections in host populations. Here, we report a previously unrecognized nutritional aspect, the provision of riboflavin (vitamin B2), that potentially underpins the Wolbachia-mediated fitness benefit to insect hosts. A comparative genomic survey for synthetic capability of B vitamins revealed that only the synthesis pathway for riboflavin is highly conserved among diverse insect-associated Wolbachia strains, while the synthesis pathways for other B vitamins were either incomplete or absent. Molecular phylogenetic and genomic analyses of riboflavin synthesis genes from diverse Wolbachia strains revealed that, in general, their phylogenetic relationships are concordant with Wolbachia's genomic phylogeny, suggesting that the riboflavin synthesis genes have been stably maintained in the course of Wolbachia evolution. In rearing experiments with bedbugs (Cimex lectularius) on blood meals in which B vitamin contents were manipulated, we demonstrated that Wolbachia's riboflavin provisioning significantly contributes to growth, survival, and reproduction of the insect host. These results provide a physiological basis upon which Wolbachia-mediated positive fitness consequences are manifested and shed new light on the ecological and evolutionary relevance of Wolbachia infections.

IMPORTANCE: Conventionally, Wolbachia has been regarded as a parasitic bacterial endosymbiont that manipulates the host insect's reproduction in a selfish manner, which tends to affect a host's fitness negatively. Meanwhile, some theories predict that, at the same time, Wolbachia can directly affect the host's fitness positively, which may potentially reconcile the negative effect and facilitate spread and stability of the symbiotic association. Here we demonstrate, by using comparative genomic and experimental approaches, that among synthetic pathways for B vitamins, the synthetic pathway for riboflavin (vitamin B2) is exceptionally conserved among diverse insect-associated Wolbachia strains, and Wolbachia's riboflavin provisioning certainly contributes to growth, survival, and reproduction in an insect. These findings uncover a nutritional mechanism of a Wolbachia-mediated fitness benefit, which provides empirical evidence highlighting a "Jekyll and Hyde" aspect of Wolbachia infection.}, } @article {pmid26548557, year = {2016}, author = {Arbuthnott, D and Levin, TC and Promislow, DE}, title = {The impacts of Wolbachia and the microbiome on mate choice in Drosophila melanogaster.}, journal = {Journal of evolutionary biology}, volume = {29}, number = {2}, pages = {461-468}, pmid = {26548557}, issn = {1420-9101}, support = {R01 GM102279/GM/NIGMS NIH HHS/United States ; GM102279/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*microbiology/*physiology ; Female ; Male ; Mating Preference, Animal/*physiology ; Microbiota/*physiology ; Species Specificity ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Symbionts and parasites can manipulate their hosts' reproduction to their own benefit, profoundly influencing patterns of mate choice and evolution of the host population. Wolbachia is one of the most widespread symbionts among arthropods, and one that alters its hosts' reproduction in diverse and dramatic ways. While we are beginning to appreciate how Wolbachia's extreme manipulations of host reproduction can influence species diversification and reproductive isolation, we understand little about how symbionts and Wolbachia, in particular, may affect intrapopulation processes of mate choice. We hypothesized that the maternally transmitted Wolbachia would increase the attractiveness of its female hosts to further its own spread. We therefore tested the effects of Wolbachia removal and microbiome disruption on female attractiveness and male mate choice among ten isofemale lines of Drosophila melanogaster. We found variable effects of general microbiome disruption on female attractiveness, with indications that bacteria interact with hosts in a line-specific manner to affect female attractiveness. However, we found no evidence that Wolbachia influence female attractiveness or male mate choice among these lines. Although the endosymbiont Wolbachia can greatly alter the reproduction of their hosts in many species, there is no indication that they alter mate choice behaviours in D. melanogaster.}, } @article {pmid26542076, year = {2015}, author = {Lawrence, AL and Hii, SF and Chong, R and Webb, CE and Traub, R and Brown, G and Šlapeta, J}, title = {Evaluation of the bacterial microbiome of two flea species using different DNA-isolation techniques provides insights into flea host ecology.}, journal = {FEMS microbiology ecology}, volume = {91}, number = {12}, pages = {}, doi = {10.1093/femsec/fiv134}, pmid = {26542076}, issn = {1574-6941}, mesh = {Animals ; Australia ; Bacteroidetes/genetics/*isolation & purification ; Base Sequence ; Cats ; Cyclooxygenase 1/genetics ; DNA, Bacterial/genetics/*isolation & purification ; High-Throughput Nucleotide Sequencing ; *Host-Pathogen Interactions ; Microbiota/*genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/*isolation & purification ; Sequence Analysis, DNA ; Siphonaptera/*microbiology ; Tachyglossidae/microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Fleas (Siphonaptera) are ubiquitous blood-sucking pests of animals worldwide and are vectors of zoonotic bacteria such as Rickettsia and Bartonella. We performed Ion Torrent PGM amplicon sequencing for the bacterial 16S rRNA gene to compare the microbiome of the ubiquitous cat flea (Ctenocephalides f. felis) and the host-specific echidna stickfast flea (Echidnophaga a. ambulans) and evaluated potential bias produced during common genomic DNA-isolation methods. We demonstrated significant differences in the bacterial community diversity between the two flea species but not between protocols combining surface sterilisation with whole flea homogenisation or exoskeleton retention. Both flea species were dominated by obligate intracellular endosymbiont Wolbachia, and the echidna stickfast fleas possessed the endosymbiont Cardinium. Cat fleas that were not surface sterilised showed presence of Candidatus 'Rickettsia senegalensis' DNA, the first report of its presence in Australia. In the case of Rickettsia, we show that sequencing depth of 50 000 was required for comparable sensitivity with Rickettsia qPCR. Low-abundance bacterial genera are suggested to reflect host ecology. The deep-sequencing approach demonstrates feasibility of pathogen detection with simultaneous quantitative analysis and evaluation of the inter-relationship of microbes within vectors.}, } @article {pmid26538596, year = {2015}, author = {Bernhauerová, V and Berec, L and Maxin, D}, title = {Evolution of early male-killing in horizontally transmitted parasites.}, journal = {Proceedings. Biological sciences}, volume = {282}, number = {1818}, pages = {20152068}, pmid = {26538596}, issn = {1471-2954}, mesh = {Animals ; Bacteria/classification/*genetics ; Bacterial Physiological Phenomena ; *Biological Evolution ; *Disease Transmission, Infectious ; Female ; *Host-Pathogen Interactions ; Insecta/embryology/*microbiology ; Male ; Sex Ratio ; }, abstract = {Early male-killing (MK) bacteria are vertically transmitted reproductive parasites which kill male offspring that inherit them. Whereas their incidence is well documented, characteristics allowing originally non-MK bacteria to gradually evolve MK ability remain unclear. We show that horizontal transmission is a mechanism enabling vertically transmitted bacteria to evolve fully efficient MK under a wide range of host and parasite characteristics, especially when the efficacy of vertical transmission is high. We also show that an almost 100% vertically transmitted and 100% effective male-killer may evolve from a purely horizontally transmitted non-MK ancestor, and that a 100% efficient male-killer can form a stable coexistence only with a non-MK bacterial strain. Our findings are in line with the empirical evidence on current MK bacteria, explain their high efficacy in killing infected male embryos and their variability within and across insect taxa, and suggest that they may have evolved independently in phylogenetically distinct species.}, } @article {pmid26531251, year = {2016}, author = {Yeap, HL and Rašić, G and Endersby-Harshman, NM and Lee, SF and Arguni, E and Le Nguyen, H and Hoffmann, AA}, title = {Mitochondrial DNA variants help monitor the dynamics of Wolbachia invasion into host populations.}, journal = {Heredity}, volume = {116}, number = {3}, pages = {265-276}, pmid = {26531251}, issn = {1365-2540}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Australia ; DNA, Mitochondrial/*genetics ; Genetic Markers ; *Genetic Variation ; Genetics, Population ; Haplotypes ; Insect Vectors/genetics/microbiology ; Models, Biological ; Models, Genetic ; *Pest Control, Biological ; *Wolbachia ; }, abstract = {Wolbachia is the most widespread endosymbiotic bacterium of insects and other arthropods that can rapidly invade host populations. Deliberate releases of Wolbachia into natural populations of the dengue fever mosquito, Aedes aegypti, are used as a novel biocontrol strategy for dengue suppression. Invasion of Wolbachia through the host population relies on factors such as high fidelity of the endosymbiont transmission and limited immigration of uninfected individuals, but these factors can be difficult to measure. One way of acquiring relevant information is to consider mitochondrial DNA (mtDNA) variation alongside Wolbachia in field-caught mosquitoes. Here we used diagnostic mtDNA markers to differentiate infection-associated mtDNA haplotypes from those of the uninfected mosquitoes at release sites. Unique haplotypes associated with Wolbachia were found at locations outside Australia. We also performed mathematical and qualitative analyses including modelling the expected dynamics of the Wolbachia and mtDNA variants during and after a release. Our analyses identified key features in haplotype frequency patterns to infer the presence of imperfect maternal transmission of Wolbachia, presence of immigration and possibly incomplete cytoplasmic incompatibility. We demonstrate that ongoing screening of the mtDNA variants should provide information on maternal leakage and immigration, particularly in releases outside Australia. As we demonstrate in a case study, our models to track the Wolbachia dynamics can be successfully applied to temporal studies in natural populations or Wolbachia release programs, as long as there is co-occurring mtDNA variation that differentiates infected and uninfected populations.}, } @article {pmid26510523, year = {2015}, author = {Nguyen, TH and Nguyen, HL and Nguyen, TY and Vu, SN and Tran, ND and Le, TN and Vien, QM and Bui, TC and Le, HT and Kutcher, S and Hurst, TP and Duong, TT and Jeffery, JA and Darbro, JM and Kay, BH and Iturbe-Ormaetxe, I and Popovici, J and Montgomery, BL and Turley, AP and Zigterman, F and Cook, H and Cook, PE and Johnson, PH and Ryan, PA and Paton, CJ and Ritchie, SA and Simmons, CP and O'Neill, SL and Hoffmann, AA}, title = {Field evaluation of the establishment potential of wMelPop Wolbachia in Australia and Vietnam for dengue control.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {563}, pmid = {26510523}, issn = {1756-3305}, mesh = {Aedes/*microbiology/physiology ; Animals ; Australia/epidemiology ; Dengue/*prevention & control/transmission ; Environment ; Female ; Humans ; Insect Vectors/*microbiology ; Male ; Pest Control, Biological/*methods ; Pupa ; Vietnam/epidemiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Introduced Wolbachia bacteria can influence the susceptibility of Aedes aegypti mosquitoes to arboviral infections as well as having detrimental effects on host fitness. Previous field trials demonstrated that the wMel strain of Wolbachia effectively and durably invades Ae. aegypti populations. Here we report on trials of a second strain, wMelPop-PGYP Wolbachia, in field sites in northern Australia (Machans Beach and Babinda) and central Vietnam (Tri Nguyen, Hon Mieu Island), each with contrasting natural Ae. aegypti densities.

METHODS: Mosquitoes were released at the adult or pupal stages for different lengths of time at the sites depending on changes in Wolbachia frequency as assessed through PCR assays of material collected through Biogents-Sentinel (BG-S) traps and ovitraps. Adult numbers were also monitored through BG-S traps. Changes in Wolbachia frequency were compared across hamlets or house blocks.

RESULTS: Releases of adult wMelPop-Ae. aegypti resulted in the transient invasion of wMelPop in all three field sites. Invasion at the Australian sites was heterogeneous, reflecting a slower rate of invasion in locations where background mosquito numbers were high. In contrast, invasion across Tri Nguyen was relatively uniform. After cessation of releases, the frequency of wMelPop declined in all sites, most rapidly in Babinda and Tri Nguyen. Within Machans Beach the rate of decrease varied among areas, and wMelPop was detected for several months in an area with a relatively low mosquito density.

CONCLUSIONS: These findings highlight challenges associated with releasing Wolbachia-Ae. aegypti combinations with low fitness, albeit strong virus interference properties, as a means of sustainable control of dengue virus transmission.}, } @article {pmid26501838, year = {2016}, author = {Pathak, M and Sharma, P and Sharma, A and Verma, M and Srivastava, M and Misra-Bhattacharya, S}, title = {Regulatory T-cell neutralization in mice during filariasis helps in parasite clearance by enhancing T helper type 17-mediated pro-inflammatory response.}, journal = {Immunology}, volume = {147}, number = {2}, pages = {190-203}, pmid = {26501838}, issn = {1365-2567}, mesh = {Animals ; Antibodies, Neutralizing/*pharmacology ; Bacterial Outer Membrane Proteins/immunology ; CD40 Antigens/immunology/metabolism ; Cells, Cultured ; Dendritic Cells/drug effects/immunology/parasitology ; Disease Models, Animal ; Elephantiasis, Filarial/*drug therapy/immunology/metabolism/parasitology ; Eosinophils/drug effects/immunology/parasitology ; Glucocorticoid-Induced TNFR-Related Protein/*antagonists & inhibitors/immunology/metabolism ; Host-Parasite Interactions ; Immunization ; Inflammation Mediators/*immunology/metabolism ; Interferon-gamma/immunology/metabolism ; Interleukin-10/immunology/metabolism ; Interleukin-2 Receptor alpha Subunit/*antagonists & inhibitors/immunology/metabolism ; Macrophage Activation/drug effects ; Mice, Inbred BALB C ; T-Lymphocytes, Regulatory/*drug effects/immunology/metabolism/parasitology ; Th17 Cells/*drug effects/immunology/metabolism/parasitology ; Time Factors ; }, abstract = {Lymphatic filariasis leads to profound impairment of parasite-specific T helper type 1 (Th1) and Th2 immune responses and significantly increases the expression of regulatory networks and regulatory effectors like transforming growth factor-β, CD25, cytotoxic T-lymphocyte antigen 4, glucocorticoid-induced tumour necrosis factor receptor (GITR) and regulatory T (Treg) cells, which together play an important role in immunosuppression. While Treg cells suppress the activity of effector cells, monocyte dysfunction, characterized by an alternatively activated immunoregulatory phenotype, is one hypothesis that explains the lack of an antigen-specific T-cell response in infected individuals. In the present study, we administered neutralizing antibodies against the Treg cell-associated markers CD25 and GITR and observed its effects on filaria-induced immunosuppression. Our results show that administration of anti-CD25 and anti-GITR in infected animals not only arrested the accumulation of Treg cells and reduced arginase activity, but also led to an increase in the percentages of Th17 cells in the secondary lymphoid organs of mice. Elevated levels of interferon-γ and decreased levels of interleukin-10 were also noted in the culture supernatants of mouse splenocytes that were treated with neutralizing antibodies. Furthermore, treatment with neutralizing antibodies enhanced the expression of inducible nitric oxide synthase on host macrophages and CD40 on host dendritic cells with concomitant decreased expression of alternative activation markers Arg1, Ym1 and Fizz1, which together lead to reduced parasite burden in treated animals. In summary, administration of neutralizing antibodies helps in breaking the regulatory network in mice and limits parasite-induced immunosuppression at the earliest host-parasite interface.}, } @article {pmid26499484, year = {2015}, author = {Agamennone, V and Jakupović, D and Weedon, JT and Suring, WJ and van Straalen, NM and Roelofs, D and Röling, WF}, title = {The microbiome of Folsomia candida: an assessment of bacterial diversity in a Wolbachia-containing animal.}, journal = {FEMS microbiology ecology}, volume = {91}, number = {11}, pages = {}, doi = {10.1093/femsec/fiv128}, pmid = {26499484}, issn = {1574-6941}, mesh = {Animals ; Arthropods/*microbiology/physiology ; Bacteria/*classification/genetics/isolation & purification ; Bacterial Physiological Phenomena ; DNA, Bacterial/genetics ; Denaturing Gradient Gel Electrophoresis ; Microbiota ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA/methods ; *Soil ; Soil Microbiology ; Symbiosis ; Wolbachia/physiology ; }, abstract = {The springtail Folsomia candida is an important model organism for soil ecology, ecotoxicology and ecogenomics. The decomposer activities of soil invertebrates like Folsomia depend on their relationship with microbial communities including gut symbionts. In this paper, we apply high-throughput sequencing to provide a detailed characterization of the bacterial community associated with parthenogenetic F. candida. First, we evaluated a method to suppress the amplification of DNA from the endosymbiont Wolbachia, to prevent it from interfering with the identification of less abundant operational taxonomic units (OTUs). The suppression treatment applied was effective against Wolbachia and did not interfere with the detection of the most abundant OTUs (59 OTUs, contributing over 87% of the reads). However, this method did affect the inferred community composition. Significant differences were subsequently observed in the composition of bacterial communities associated with two different strains of F. candida. A total of 832 OTUs were found, of which 45% were only present in one strain and 17% only in the other. Among the 20 most abundant OTUs, 16 were shared between strains. Denaturing gradient gel electrophoresis and clone libraries, although unable to capture the full diversity of the bacterial community, provided results that supported the NGS data.}, } @article {pmid26497146, year = {2015}, author = {Gutzwiller, F and Carmo, CR and Miller, DE and Rice, DW and Newton, IL and Hawley, RS and Teixeira, L and Bergman, CM}, title = {Dynamics of Wolbachia pipientis Gene Expression Across the Drosophila melanogaster Life Cycle.}, journal = {G3 (Bethesda, Md.)}, volume = {5}, number = {12}, pages = {2843-2856}, pmid = {26497146}, issn = {2160-1836}, support = {BB/L002817/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Cluster Analysis ; Computational Biology ; Drosophila melanogaster/*growth & development/microbiology ; Gene Expression Profiling ; *Gene Expression Regulation, Bacterial ; Genome, Bacterial ; High-Throughput Nucleotide Sequencing ; Host-Pathogen Interactions/genetics ; Life Cycle Stages ; Phylogeny ; Stress, Physiological/genetics ; Symbiosis ; Transcriptome ; Wolbachia/*genetics ; }, abstract = {Symbiotic interactions between microbes and their multicellular hosts have manifold biological consequences. To better understand how bacteria maintain symbiotic associations with animal hosts, we analyzed genome-wide gene expression for the endosymbiotic α-proteobacteria Wolbachia pipientis across the entire life cycle of Drosophila melanogaster. We found that the majority of Wolbachia genes are expressed stably across the D. melanogaster life cycle, but that 7.8% of Wolbachia genes exhibit robust stage- or sex-specific expression differences when studied in the whole-organism context. Differentially-expressed Wolbachia genes are typically up-regulated after Drosophila embryogenesis and include many bacterial membrane, secretion system, and ankyrin repeat-containing proteins. Sex-biased genes are often organized as small operons of uncharacterized genes and are mainly up-regulated in adult Drosophila males in an age-dependent manner. We also systematically investigated expression levels of previously-reported candidate genes thought to be involved in host-microbe interaction, including those in the WO-A and WO-B prophages and in the Octomom region, which has been implicated in regulating bacterial titer and pathogenicity. Our work provides comprehensive insight into the developmental dynamics of gene expression for a widespread endosymbiont in its natural host context, and shows that public gene expression data harbor rich resources to probe the functional basis of the Wolbachia-Drosophila symbiosis and annotate the transcriptional outputs of the Wolbachia genome.}, } @article {pmid26496201, year = {2015}, author = {Binnebose, AM and Haughney, SL and Martin, R and Imerman, PM and Narasimhan, B and Bellaire, BH}, title = {Polyanhydride Nanoparticle Delivery Platform Dramatically Enhances Killing of Filarial Worms.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {10}, pages = {e0004173}, pmid = {26496201}, issn = {1935-2735}, support = {R01 AI047195/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anthelmintics/*pharmacology ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/*drug effects/*physiology ; Doxycycline/pharmacology ; Drug Carriers/*metabolism ; Ivermectin/pharmacology ; Locomotion/drug effects ; Nanoparticles/*metabolism ; Parasitic Sensitivity Tests ; Polyanhydrides/*metabolism ; Survival Analysis ; }, abstract = {Filarial diseases represent a significant social and economic burden to over 120 million people worldwide and are caused by endoparasites that require the presence of symbiotic bacteria of the genus Wolbachia for fertility and viability of the host parasite. Targeting Wolbachia for elimination is a therapeutic approach that shows promise in the treatment of onchocerciasis and lymphatic filariasis. Here we demonstrate the use of a biodegradable polyanhydride nanoparticle-based platform for the co-delivery of the antibiotic doxycycline with the antiparasitic drug, ivermectin, to reduce microfilarial burden and rapidly kill adult worms. When doxycycline and ivermectin were co-delivered within polyanhydride nanoparticles, effective killing of adult female Brugia malayi filarial worms was achieved with approximately 4,000-fold reduction in the amount of drug used. Additionally the time to death of the macrofilaria was also significantly reduced (five-fold) when the anti-filarial drug cocktail was delivered within polyanhydride nanoparticles. We hypothesize that the mechanism behind this dramatically enhanced killing of the macrofilaria is the ability of the polyanhydride nanoparticles to behave as a Trojan horse and penetrate the cuticle, bypassing excretory pumps of B. malayi, and effectively deliver drug directly to both the worm and Wolbachia at high enough microenvironmental concentrations to cause death. These provocative findings may have significant consequences for the reduction in the amount of drug and the length of treatment required for filarial infections in terms of patient compliance and reduced cost of treatment.}, } @article {pmid26495042, year = {2015}, author = {Rašić, G and Schama, R and Powell, R and Maciel-de Freitas, R and Endersby-Harshman, NM and Filipović, I and Sylvestre, G and Máspero, RC and Hoffmann, AA}, title = {Contrasting genetic structure between mitochondrial and nuclear markers in the dengue fever mosquito from Rio de Janeiro: implications for vector control.}, journal = {Evolutionary applications}, volume = {8}, number = {9}, pages = {901-915}, pmid = {26495042}, issn = {1752-4571}, abstract = {Dengue is the most prevalent global arboviral disease that affects over 300 million people every year. Brazil has the highest number of dengue cases in the world, with the most severe epidemics in the city of Rio de Janeiro (Rio). The effective control of dengue is critically dependent on the knowledge of population genetic structuring in the primary dengue vector, the mosquito Aedes aegypti. We analyzed mitochondrial and nuclear genomewide single nucleotide polymorphism markers generated via Restriction-site Associated DNA sequencing, as well as traditional microsatellite markers in Ae. aegypti from Rio. We found four divergent mitochondrial lineages and a strong spatial structuring of mitochondrial variation, in contrast to the overall nuclear homogeneity across Rio. Despite a low overall differentiation in the nuclear genome, we detected strong spatial structure for variation in over 20 genes that have a significantly altered expression in response to insecticides, xenobiotics, and pathogens, including the novel biocontrol agent Wolbachia. Our results indicate that high genetic diversity, spatially unconstrained admixing likely mediated by male dispersal, along with locally heterogeneous genetic variation that could affect insecticide resistance and mosquito vectorial capacity, set limits to the effectiveness of measures to control dengue fever in Rio.}, } @article {pmid26492411, year = {2015}, author = {Furihata, S and Hirata, M and Matsumoto, H and Hayakawa, Y}, title = {Bacteria Endosymbiont, Wolbachia, Promotes Parasitism of Parasitoid Wasp Asobara japonica.}, journal = {PloS one}, volume = {10}, number = {10}, pages = {e0140914}, pmid = {26492411}, issn = {1932-6203}, mesh = {Animals ; Drosophila melanogaster/parasitology ; Female ; Host-Pathogen Interactions/physiology ; Male ; Oviposition/physiology ; Symbiosis/physiology ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is the most widespread endosymbiotic bacterium that manipulates reproduction of its arthropod hosts to enhance its own spread throughout host populations. Infection with Wolbachia causes complete parthenogenetic reproduction in many Hymenoptera, producing only female offspring. The mechanism of such reproductive manipulation by Wolbachia has been extensively studied. However, the effects of Wolbachia symbiosis on behavioral traits of the hosts are scarcely investigated. The parasitoid wasp Asobara japonica is an ideal insect to investigate this because symbiotic and aposymbiotic strains are available: Wolbachia-infected Tokyo (TK) and noninfected Iriomote (IR) strains originally collected on the main island and southwest islands of Japan, respectively. We compared the oviposition behaviors of the two strains and found that TK strain females parasitized Drosophila melanogaster larvae more actively than the IR strain, especially during the first two days after eclosion. Removing Wolbachia from the TK strain wasps by treatment with tetracycline or rifampicin decreased their parasitism activity to the level of the IR strain. Morphological and behavioral analyses of both strain wasps showed that Wolbachia endosymbionts do not affect development of the host female reproductive tract and eggs, but do enhance host-searching ability of female wasps. These results suggest the possibility that Wolbachia endosymbionts may promote their diffusion and persistence in the host A. japonica population not only at least partly by parthenogenesis but also by enhancement of oviposition frequency of the host females.}, } @article {pmid26485147, year = {2015}, author = {Jones, RT and Borchert, J and Eisen, R and MacMillan, K and Boegler, K and Gage, KL}, title = {Flea-Associated Bacterial Communities across an Environmental Transect in a Plague-Endemic Region of Uganda.}, journal = {PloS one}, volume = {10}, number = {10}, pages = {e0141057}, pmid = {26485147}, issn = {1932-6203}, mesh = {Actinobacteria/genetics/isolation & purification ; Animals ; Bacteroidetes/genetics/isolation & purification ; Base Sequence ; DNA, Bacterial/genetics ; Firmicutes/genetics/isolation & purification ; Humans ; Insect Vectors/*microbiology ; Microbial Consortia/*genetics ; Plague/*microbiology ; Proteobacteria/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Rodentia/*microbiology ; Sequence Analysis, DNA ; Siphonaptera/*microbiology ; Uganda ; Yersinia pestis/pathogenicity ; }, abstract = {The vast majority of human plague cases currently occur in sub-Saharan Africa. The primary route of transmission of Yersinia pestis, the causative agent of plague, is via flea bites. Non-pathogenic flea-associated bacteria may interact with Y. pestis within fleas and it is important to understand what factors govern flea-associated bacterial assemblages. Six species of fleas were collected from nine rodent species from ten Ugandan villages between October 2010 and March 2011. A total of 660,345 16S rRNA gene DNA sequences were used to characterize bacterial communities of 332 individual fleas. The DNA sequences were binned into 421 Operational Taxonomic Units (OTUs) based on 97% sequence similarity. We used beta diversity metrics to assess the effects of flea species, flea sex, rodent host species, site (i.e. village), collection date, elevation, mean annual precipitation, average monthly precipitation, and average monthly temperature on bacterial community structure. Flea species had the greatest effect on bacterial community structure with each flea species harboring unique bacterial lineages. The site (i.e. village), rodent host, flea sex, elevation, precipitation, and temperature also significantly affected bacterial community composition. Some bacterial lineages were widespread among flea species (e.g. Bartonella spp. and Wolbachia spp.), but each flea species also harbored unique bacterial lineages. Some of these lineages are not closely related to known bacterial diversity and likely represent newly discovered lineages of insect symbionts. Our finding that flea species has the greatest effect on bacterial community composition may help future investigations between Yersinia pestis and non-pathogenic flea-associated bacteria. Characterizing bacterial communities of fleas during a plague epizootic event in the future would be helpful.}, } @article {pmid26470291, year = {2015}, author = {Mattio, MF and Argüello Caro, EB and Rodriguero, MS and Dumón, AD and Alemandri, VM and Truol, G}, title = {Wolbachia Occurrence in Planthopper (Hemiptera: Delphacidae) Vectors of Cereal Viruses in Argentina.}, journal = {Journal of economic entomology}, volume = {108}, number = {4}, pages = {1526-1530}, doi = {10.1093/jee/tov140}, pmid = {26470291}, issn = {0022-0493}, mesh = {Animals ; Argentina ; Bacterial Proteins/genetics ; DNA, Bacterial/genetics ; Hemiptera/*microbiology/virology ; Insect Vectors/*microbiology/virology ; Molecular Sequence Data ; Plant Diseases/virology ; Plant Viruses/physiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Triticum/virology ; Wolbachia/*genetics/*isolation & purification/physiology ; Zea mays/virology ; }, abstract = {Maize (Zea mays L.) and wheat (Triticum aestivum L.) are the most important cereal crops for the Argentinean economy and are affected by several diseases. Different planthopper species transmit causal agents of some of those diseases, including Mal de Río Cuarto virus, barley yellow striate mosaic virus, and the recently proposed maize yellow striate virus. Many planthopper species are sap feeders and therefore are expected to host bacteria providing essential nutrients lacking in the diet. Previous studies have evidenced that some of these bacterial symbionts are involved in the virus transmission. Wolbachia is a group of obligate intracellular bacteria infecting numerous arthropod species and causing reproductive alterations in their hosts. These bacteria have been detected in planthopper species, considered rice pests in various regions of the world. To date, Wolbachia infection status of planthopper species of Argentina is unknown. Amplification by PCR and sequencing of 16S rDNA, wsp- and ftsZ-specific genes demonstrated Wolbachia infection in Caenodelphax teapae (Fowler), Delphacodes kuscheli Fennah, Pyrophagus tigrinus Remes Lenicov & Varela, Tagosodes orizicolus (Muir), and Toya propinqua (Fieber). This is the first report of Wolbachia in delphacid vectors of viruses affecting maize and wheat. An understanding of the bacterial diversity harbored by these insect vectors could lead to new options for future management of diseases of economically important crops in a developing country.}, } @article {pmid26470269, year = {2015}, author = {Ming, QL and Shen, JF and Cheng, C and Liu, CM and Feng, ZJ}, title = {Wolbachia Infection Dynamics in Tribolium confusum (Coleoptera: Tenebrionidae) and Their Effects on Host Mating Behavior and Reproduction.}, journal = {Journal of economic entomology}, volume = {108}, number = {3}, pages = {1408-1415}, doi = {10.1093/jee/tov053}, pmid = {26470269}, issn = {0022-0493}, mesh = {Animals ; Female ; Larva/growth & development/microbiology ; Male ; Ovum/growth & development/microbiology ; Pupa/growth & development/microbiology ; Real-Time Polymerase Chain Reaction ; Reproduction ; *Sexual Behavior, Animal ; Tribolium/growth & development/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia interact with their hosts in a broad variety of relationships that range from parasitism to mutualism. To improve the understanding of complex relationships between Wolbachia and host, we performed not only mating and crossing experiments to investigate effects of Wolbachia on mate choice, mating performance, and reproduction in the confused flour beetles Tribolium confusum (Jacquelin du Val), but also quantitative PCR to determine Wolbachia spatiotemporal infection density dynamics within beetles. Wolbachia induced strong cytoplasmic incompatibility, but had no effects on male mate choice and mating performance. Compared with Wolbachia-uninfected females, infected females had very high fecundity irrespective of male's infection status. Wolbachia infection densities in beetles were higher in eggs and adults and in the reproductive tissues and abdomens, whereas Wolbachia density in adults did not differ between sexes and among different ages. These results suggest that Wolbachia have evolved mutualistic interactions with T. confusum, which provides the first evidence of Wolbachia mutualisms in this beetle species. We discussed these findings and their evolutionary implications in light of Wolbachia-host interactions.}, } @article {pmid26470122, year = {2015}, author = {Ji, HL and Qi, LD and Hong, XY and Xie, HF and Li, YX}, title = {Effects of Host Sex, Plant Species, and Putative Host Species on the Prevalence of Wolbachia in Natural Populations of Bemisia tabaci (Hemiptera: Aleyrodidae): A Modified Nested PCR Study.}, journal = {Journal of economic entomology}, volume = {108}, number = {1}, pages = {210-218}, doi = {10.1093/jee/tou004}, pmid = {26470122}, issn = {0022-0493}, mesh = {Animals ; *Crops, Agricultural ; Female ; Hemiptera/*microbiology ; Male ; Polymerase Chain Reaction ; Wolbachia/*genetics ; }, abstract = {Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a globally distributed pest. One of the key endosymbionts in B. tabaci is Wolbachia, an α-proteobacterium implicated in many important biological processes. Previous studies indicated that the infection frequency of Wolbachia in Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) varied greatly among populations in different areas. However, little is known about the factors that influence the prevalence of Wolbachia in B. tabaci. In this paper, 25 field populations were collected from different locations in China, and 1,161 individuals were screened for the presence of Wolbachia using a nested polymerase chain reaction (PCR)-based method, which targets the wsp gene, to confirm Wolbachia infection status. The prevalence of Wolbachia ranged from 1.54 to 66.67% within the 25 field populations, and the infection frequency of Wolbachia was affected significantly by the putative species of B. tabaci. The infection frequency (51.55%) of Wolbachia was significantly greater in native species than in the MED (25.65%) and MEAM1 (14.37%). With the exception of host plant, all factors, including putative species, geographic location, and the sex of the host, affected the Wolbachia infection frequency in whiteflies. Six Wolbachia strains were found and clustered into four distinct clades upon phylogenetic analyses. Furthermore, Wolbachia in B. tabaci have close relationships with those from other host species, including Liriomyza trifolii (Burgess), Sogatella furcifera (Horvath), Nilaparvata lugens (Stål), and Culex pipiens L. The results demonstrated the variation and diversity of Wolbachia in B. tabaci field populations, and that the application of nested PCR extended our knowledge of Wolbachia infection in B. tabaci, especially in invasive whiteflies.}, } @article {pmid26463408, year = {2015}, author = {Wuliandari, JR and Lee, SF and White, VL and Tantowijoyo, W and Hoffmann, AA and Endersby-Harshman, NM}, title = {Association between Three Mutations, F1565C, V1023G and S996P, in the Voltage-Sensitive Sodium Channel Gene and Knockdown Resistance in Aedes aegypti from Yogyakarta, Indonesia.}, journal = {Insects}, volume = {6}, number = {3}, pages = {658-685}, pmid = {26463408}, issn = {2075-4450}, abstract = {Mutations in the voltage-sensitive sodium channel gene (Vssc) have been identified in Aedes aegypti and some have been associated with pyrethroid insecticide resistance. Whether these mutations cause resistance, alone or in combination with other alleles, remains unclear, but must be understood if mutations are to become markers for resistance monitoring. We describe High Resolution Melt (HRM) genotyping assays for assessing mutations found in Ae. aegypti in Indonesia (F1565C, V1023G, S996P) and use them to test for associations with pyrethroid resistance in mosquitoes from Yogyakarta, a city where insecticide use is widespread. Such knowledge is important because Yogyakarta is a target area for releases of Wolbachia-infected mosquitoes with virus-blocking traits for dengue suppression. We identify three alleles across Yogyakarta putatively linked to resistance in previous research. By comparing resistant and susceptible mosquitoes from bioassays, we show that the 1023G allele is associated with resistance to type I and type II pyrethroids. In contrast, F1565C homozygotes were rare and there was only a weak association between individuals heterozygous for the mutation and resistance to a type I pyrethroid. As the heterozygote is expected to be incompletely recessive, it is likely that this association was due to a different resistance mechanism being present. A resistance advantage conferred to V1023G homozygotes through addition of the S996P allele in the homozygous form was suggested for the Type II pyrethroid, deltamethrin. Screening of V1023G and S996P should assist resistance monitoring in Ae. aegypti from Yogyakarta, and these mutations should be maintained in Wolbachia strains destined for release in this city to ensure that these virus-blocking strains of mosquitoes are not disadvantaged, relative to resident populations.}, } @article {pmid26463078, year = {2015}, author = {Kean, J and Rainey, SM and McFarlane, M and Donald, CL and Schnettler, E and Kohl, A and Pondeville, E}, title = {Fighting Arbovirus Transmission: Natural and Engineered Control of Vector Competence in Aedes Mosquitoes.}, journal = {Insects}, volume = {6}, number = {1}, pages = {236-278}, pmid = {26463078}, issn = {2075-4450}, support = {//Wellcome Trust/United Kingdom ; MC_UP_A550_1031/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, abstract = {Control of aedine mosquito vectors, either by mosquito population reduction or replacement with refractory mosquitoes, may play an essential role in the fight against arboviral diseases. In this review, we will focus on the development and application of biological approaches, both natural or engineered, to limit mosquito vector competence for arboviruses. The study of mosquito antiviral immunity has led to the identification of a number of host response mechanisms and proteins that are required to control arbovirus replication in mosquitoes, though more factors influencing vector competence are likely to be discovered. We will discuss key aspects of these pathways as targets either for selection of naturally resistant mosquito populations or for mosquito genetic manipulation. Moreover, we will consider the use of endosymbiotic bacteria such as Wolbachia, which in some cases have proven to be remarkably efficient in disrupting arbovirus transmission by mosquitoes, but also the use of naturally occurring insect-specific viruses that may interfere with arboviruses in mosquito vectors. Finally, we will discuss the use of paratransgenesis as well as entomopathogenic fungi, which are also proposed strategies to control vector competence.}, } @article {pmid26453817, year = {2015}, author = {Sugimoto, TN and Kayukawa, T and Shinoda, T and Ishikawa, Y and Tsuchida, T}, title = {Misdirection of dosage compensation underlies bidirectional sex-specific death in Wolbachia-infected Ostrinia scapulalis.}, journal = {Insect biochemistry and molecular biology}, volume = {66}, number = {}, pages = {72-76}, doi = {10.1016/j.ibmb.2015.10.001}, pmid = {26453817}, issn = {1879-0240}, mesh = {Animals ; Death ; *Dosage Compensation, Genetic ; Female ; Genotype ; Larva/genetics/growth & development/microbiology ; Male ; Moths/*genetics/growth & development/*microbiology ; Sex Chromosomes ; Sex Differentiation ; Sex Ratio ; Tetracycline/pharmacology ; Wolbachia/*physiology ; }, abstract = {Endosymbiotic bacteria of the genus Wolbachia often manipulate the reproductive system of their hosts to propagate themselves in host populations. Ostrinia scapulalis moths infected with Wolbachia (wSca) produce female-only progeny (sex chromosomes: ZW), whereas females cured of the infection by antibiotic treatment produce male-only progeny (ZZ). The occurrence of female- and male-only progeny has been attributed to the specific death of the opposite sex during embryonic and larval development. In this bidirectional sex-specific lethality, embryos destined to die express a phenotypic sex opposite to their genotypic sex. On the basis of these findings, we suggested that wSca carries a genetic factor that feminizes the male host, the W chromosome of the host has lost its feminizing function, and discordance between the genotypic and phenotypic sexes underlies this sex-specific death. In the present study, we examined whether the failure of dosage compensation was responsible for this sex-specific mortality. Quantitative PCRs showed that Z-linked gene expression levels in embryos destined to die were not properly dosage compensated; they were approximately two-fold higher in the male progeny of wSca-infected females and approximately two-fold lower in the female progeny of infected-and-cured females. These results support our hypothesis that misdirection of dosage compensation underlies the sex-specific death.}, } @article {pmid26442762, year = {2015}, author = {Gutzwiller, F and Dedeine, F and Kaiser, W and Giron, D and Lopez-Vaamonde, C}, title = {Correlation between the green-island phenotype and Wolbachia infections during the evolutionary diversification of Gracillariidae leaf-mining moths.}, journal = {Ecology and evolution}, volume = {5}, number = {18}, pages = {4049-4062}, pmid = {26442762}, issn = {2045-7758}, abstract = {Internally feeding herbivorous insects such as leaf miners have developed the ability to manipulate the physiology of their host plants in a way to best meet their metabolic needs and compensate for variation in food nutritional composition. For instance, some leaf miners can induce green-islands on yellow leaves in autumn, which are characterized by photosynthetically active green patches in otherwise senescing leaves. It has been shown that endosymbionts, and most likely bacteria of the genus Wolbachia, play an important role in green-island induction in the apple leaf-mining moth Phyllonorycter blancardella. However, it is currently not known how widespread is this moth-Wolbachia-plant interaction. Here, we studied the co-occurrence between Wolbachia and the green-island phenotype in 133 moth specimens belonging to 74 species of Lepidoptera including 60 Gracillariidae leaf miners. Using a combination of molecular phylogenies and ecological data (occurrence of green-islands), we show that the acquisitions of the green-island phenotype and Wolbachia infections have been associated through the evolutionary diversification of Gracillariidae. We also found intraspecific variability in both green-island formation and Wolbachia infection, with some species being able to form green-islands without being infected by Wolbachia. In addition, Wolbachia variants belonging to both A and B supergroups were found to be associated with green-island phenotype suggesting several independent origins of green-island induction. This study opens new prospects and raises new questions about the ecology and evolution of the tripartite association between Wolbachia, leaf miners, and their host plants.}, } @article {pmid26441903, year = {2015}, author = {Minard, G and Tran, FH and Van, VT and Goubert, C and Bellet, C and Lambert, G and Kim, KL and Thuy, TH and Mavingui, P and Valiente Moro, C}, title = {French invasive Asian tiger mosquito populations harbor reduced bacterial microbiota and genetic diversity compared to Vietnamese autochthonous relatives.}, journal = {Frontiers in microbiology}, volume = {6}, number = {}, pages = {970}, pmid = {26441903}, issn = {1664-302X}, abstract = {The Asian tiger mosquito Aedes albopictus is one of the most significant pathogen vectors of the twenty-first century. Originating from Asia, it has invaded a wide range of eco-climatic regions worldwide. The insect-associated microbiota is now recognized to play a significant role in host biology. While genetic diversity bottlenecks are known to result from biological invasions, the resulting shifts in host-associated microbiota diversity has not been thoroughly investigated. To address this subject, we compared four autochthonous Ae. albopictus populations in Vietnam, the native area of Ae. albopictus, and three populations recently introduced to Metropolitan France, with the aim of documenting whether these populations display differences in host genotype and bacterial microbiota. Population-level genetic diversity (microsatellite markers and COI haplotype) and bacterial diversity (16S rDNA metabarcoding) were compared between field-caught mosquitoes. Bacterial microbiota from the whole insect bodies were largely dominated by Wolbachia pipientis. Targeted analysis of the gut microbiota revealed a greater bacterial diversity in which a fraction was common between French and Vietnamese populations. The genus Dysgonomonas was the most prevalent and abundant across all studied populations. Overall genetic diversities of both hosts and bacterial microbiota were significantly reduced in recently established populations of France compared to the autochthonous populations of Vietnam. These results open up many important avenues of investigation in order to link the process of geographical invasion to shifts in commensal and symbiotic microbiome communities, as such shifts may have dramatic impacts on the biology and/or vector competence of invading hematophagous insects.}, } @article {pmid26438427, year = {2015}, author = {Guevara-Souza, M and Vallejo, EE}, title = {A computer simulation model of Wolbachia invasion for disease vector population modification.}, journal = {BMC bioinformatics}, volume = {16}, number = {}, pages = {317}, pmid = {26438427}, issn = {1471-2105}, mesh = {Animals ; Culicidae/microbiology/physiology ; Insect Vectors/microbiology ; *Models, Theoretical ; Population Dynamics ; Reproduction ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia invasion has been proved to be a promising alternative for controlling vector-borne diseases, particularly Dengue fever. Creating computer models that can provide insight into how vector population modification can be achieved under different conditions would be most valuable for assessing the efficacy of control strategies for this disease.

METHODS: In this paper, we present a computer model that simulates the behavior of native mosquito populations after the introduction of mosquitoes infected with the Wolbachia bacteria. We studied how different factors such as fecundity, fitness cost of infection, migration rates, number of populations, population size, and number of introduced infected mosquitoes affect the spread of the Wolbachia bacteria among native mosquito populations.

RESULTS: Two main scenarios of the island model are presented in this paper, with infected mosquitoes introduced into the largest source population and peripheral populations. Overall, the results are promising; Wolbachia infection spreads among native populations and the computer model is capable of reproducing the results obtained by mathematical models and field experiments.

CONCLUSIONS: Computer models can be very useful for gaining insight into how Wolbachia invasion works and are a promising alternative for complementing experimental and mathematical approaches for vector-borne disease control.}, } @article {pmid26428255, year = {2015}, author = {Hu, L and Huang, M and Tang, M and Yu, J and Zheng, B}, title = {Wolbachia spread dynamics in stochastic environments.}, journal = {Theoretical population biology}, volume = {106}, number = {}, pages = {32-44}, doi = {10.1016/j.tpb.2015.09.003}, pmid = {26428255}, issn = {1096-0325}, mesh = {Aedes/*microbiology ; Animals ; Dengue/*prevention & control ; Extinction, Biological ; Female ; Humans ; Male ; *Models, Biological ; Population Dynamics ; Rickettsia Infections/mortality/transmission ; Stochastic Processes ; Wolbachia/*pathogenicity ; }, abstract = {Dengue fever is a mosquito-borne viral disease with 100 million people infected annually. A novel strategy for dengue control uses the bacterium Wolbachia to invade dengue vector Aedes mosquitoes. As the impact of environmental heterogeneity on Wolbachia spread dynamics in natural areas has been rarely quantified, we develop a model of differential equations for which the environmental conditions switch randomly between two regimes. We find some striking phenomena that random regime transitions could drive Wolbachia to extinction from certain initial states confirmed Wolbachia fixation in homogeneous environments, and mosquito releasing facilitates Wolbachia invasion more effectively when the regimes transit frequently. By superimposing the phase spaces of the ODE systems defined in each regime, we identify the threshold curves below which Wolbachia invades the whole population, which extends the theory of threshold infection frequency to stochastic environments.}, } @article {pmid26427709, year = {2016}, author = {Baldridge, GD and Markowski, TW and Witthuhn, BA and Higgins, L and Baldridge, AS and Fallon, AM}, title = {The Wolbachia WO bacteriophage proteome in the Aedes albopictus C/wStr1 cell line: evidence for lytic activity?.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {52}, number = {1}, pages = {77-88}, pmid = {26427709}, issn = {1543-706X}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; R01 AI 081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/microbiology ; Animals ; Bacteriophages/*genetics ; Drosophila melanogaster/*genetics/microbiology ; Gene Transfer Techniques ; Genome, Bacterial ; Hemiptera/genetics ; *Pest Control, Biological ; Phylogeny ; Proteome/*genetics ; Wolbachia/genetics/pathogenicity ; }, abstract = {Wolbachia pipientis (Rickettsiales), an obligate intracellular alphaproteobacterium in insects, manipulates host reproduction to maximize invasion of uninfected insect populations. Modification of host population structure has potential applications for control of pest species, particularly if Wolbachia can be maintained, manipulated, and genetically engineered in vitro. Although Wolbachia maintains an obligate mutualism with genome stability in nematodes, arthropods can be co-infected with distinct Wolbachia strains, and horizontal gene transfer between strains is potentially mediated by WO phages encoded within Wolbachia genomes. Proteomic analysis of a robust, persistent infection of a mosquito cell line with wStr from the planthopper, Laodelphax striatellus, revealed expression of a full array of WO phage genes, as well as nine of ten non-phage genes that occur between two distinct clusters of WOMelB genes in the genome of wMel, which infects Drosophila melanogaster. These non-phage genes encode potential host-adaptive proteins and are expressed in wStr at higher levels than phage structural proteins. A subset of seven of the non-phage genes is flanked by highly conserved non-coding sequences, including a putative promoter element, that are not present in a syntenically arranged array of homologs in plasmids from three tick-associated Rickettsia spp. These studies expand our understanding of wStr in a host cell line derived from the mosquito, Aedes albopictus, and provide a basis for investigating conditions that favor the lytic phase of the WO phage life cycle and recovery of infectious phage particles.}, } @article {pmid26423782, year = {2016}, author = {Łukasiewicz, K and Sanak, M and Węgrzyn, G}, title = {A lack of Wolbachia-specific DNA in samples from apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) individuals with deformed or reduced wings.}, journal = {Journal of applied genetics}, volume = {57}, number = {2}, pages = {271-274}, pmid = {26423782}, issn = {2190-3883}, mesh = {Animals ; Butterflies/*microbiology ; DNA, Bacterial/*isolation & purification ; Poland ; Symbiosis ; Wings, Animal/*abnormalities ; Wolbachia/*isolation & purification ; }, abstract = {Various insects contain maternally inherited endosymbiotic bacteria which can cause reproductive alterations, modulation of some physiological responses (like immunity, heat shock response, and oxidative stress response), and resistance to viral infections. In butterflies, Wolbachia sp. is the most frequent endosymbiont from this group, occurring in about 30 % of species tested to date. In this report, the presence of Wolbachia-specific DNA has been detected in apollo butterfly (Parnassius apollo). In the isolated population of this insect occurring in Pieniny National Park (Poland), malformed individuals with deformed or reduced wings appear with an exceptionally high frequency. Interestingly, while total DNA isolated from most (about 85 %) normal insects contained Wolbachia-specific sequences detected by PCR, such sequences were absent in a large fraction (70 %) of individuals with deformed wings and in all tested individuals with reduced wings. These results indicate for the first time the correlation between malformation of wings and the absence of Wolbachia sp. in insects. Although the lack of the endosymbiotic bacteria cannot be considered as the sole cause of the deformation or reduction of wings, one might suggest that Wolbachia sp. could play a protective role in the ontogenetic development of apollo butterfly.}, } @article {pmid26418658, year = {2015}, author = {Ravikumar, H and Puttaraju, HP}, title = {Molecular phylogenetic affiliation of Wolbachia and phage WO among Mansonia mosquitoes from Kerala, India.}, journal = {Journal of vector borne diseases}, volume = {52}, number = {3}, pages = {257-260}, pmid = {26418658}, issn = {0972-9062}, mesh = {Animals ; Bacteriophages/*classification/genetics/*isolation & purification ; Cluster Analysis ; Culicidae/*microbiology ; DNA, Bacterial/chemistry/genetics ; DNA, Viral/chemistry/genetics ; Female ; India ; Molecular Sequence Data ; *Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology ; Wolbachia/*classification/genetics/*isolation & purification/virology ; }, } @article {pmid26410254, year = {2015}, author = {Saeed, A and White, JA}, title = {Surveys for maternally-inherited endosymbionts reveal novel and variable infections within solitary bee species.}, journal = {Journal of invertebrate pathology}, volume = {132}, number = {}, pages = {111-114}, doi = {10.1016/j.jip.2015.09.011}, pmid = {26410254}, issn = {1096-0805}, mesh = {Animals ; Bees/*microbiology ; Enterobacteriaceae/isolation & purification/*physiology ; Gammaproteobacteria/isolation & purification/*physiology ; *Host-Pathogen Interactions ; Population Dynamics ; Symbiosis ; Wolbachia/isolation & purification/*physiology ; }, abstract = {Maternally-inherited bacteria can affect the fitness and population dynamics of their host insects; for solitary bees, such effects have the potential to influence bee efficacy as pollinators. We screened bee species for bacterial associates using 454-pyrosequencing (4 species) and diagnostic PCR (183 specimens across 29 species). The endosymbiont Wolbachia was abundant, infecting 18 species, including all specimens from the family Halictidae. Among commercially-supplied orchard bees (family Megachilidae), only 2/7 species were Wolbachia-infected, but one species showed variable infection among specimens. Two other maternally-inherited bacteria, Arsenophonus and Sodalis, were also detected, neither of which was fixed in infection frequency. Differential endosymbiont infection could potentially compromise fitness and reproductive compatibility among commercially redistributed pollinator populations.}, } @article {pmid26407882, year = {2015}, author = {Stevanovic, AL and Arnold, PA and Johnson, KN}, title = {Wolbachia-mediated antiviral protection in Drosophila larvae and adults following oral infection.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {23}, pages = {8215-8223}, pmid = {26407882}, issn = {1098-5336}, mesh = {Animals ; Antiviral Agents/*pharmacology ; Drosophila/growth & development/immunology/*microbiology/*virology ; Host-Pathogen Interactions ; Insect Viruses/*physiology ; Larva/growth & development/immunology/microbiology/virology ; Male ; Wolbachia/*physiology ; }, abstract = {Understanding viral dynamics in arthropods is of great importance when designing models to describe how viral spread can influence arthropod populations. The endosymbiotic bacterium Wolbachia spp., which is present in up to 40% of all insect species, has the ability to alter viral dynamics in both Drosophila spp. and mosquitoes, a feature that in mosquitoes may be utilized to limit spread of important arboviruses. To understand the potential effect of Wolbachia on viral dynamics in nature, it is important to consider the impact of natural routes of virus infection on Wolbachia antiviral effects. Using adult Drosophila strains, we show here that Drosophila-Wolbachia associations that have previously been shown to confer antiviral protection following systemic viral infection also confer protection against virus-induced mortality following oral exposure to Drosophila C virus in adults. Interestingly, a different pattern was observed when the same fly lines were challenged with the virus when still larvae. Analysis of the four Drosophila-Wolbachia associations that were protective in adults indicated that only the w1118-wMelPop association conferred protection in larvae following oral delivery of the virus. Analysis of Wolbachia density using quantitative PCR (qPCR) showed that a high Wolbachia density was congruent with antiviral protection in both adults and larvae. This study indicates that Wolbachia-mediated protection may vary between larval and adult stages of a given Wolbachia-host combination and that the variations in susceptibility by life stage correspond with Wolbachia density. The differences in the outcome of virus infection are likely to influence viral dynamics in Wolbachia-infected insect populations in nature and could also have important implications for the transmission of arboviruses in mosquito populations.}, } @article {pmid26407645, year = {2015}, author = {Zhang, X and Tang, S and Cheke, RA}, title = {Models to assess how best to replace dengue virus vectors with Wolbachia-infected mosquito populations.}, journal = {Mathematical biosciences}, volume = {269}, number = {}, pages = {164-177}, doi = {10.1016/j.mbs.2015.09.004}, pmid = {26407645}, issn = {1879-3134}, mesh = {Aedes/*microbiology/*virology ; Animals ; Dengue/prevention & control/transmission ; Dengue Virus/*pathogenicity ; Female ; Humans ; Insect Vectors/microbiology/virology ; Male ; Mathematical Concepts ; *Models, Biological ; Pest Control, Biological/*methods/statistics & numerical data ; Wolbachia/*physiology ; }, abstract = {Dengue fever is increasing in importance in the tropics and subtropics. Endosymbiotic Wolbachia bacteria as novel control methods can reduce the ability of virus transmission. So, many mosquitoes infected with Wolbachia are released in some countries so that strategies for population replacement can be fulfilled. However, not all of these field trails are successful, for example, releases on Tri Nguyen Island, Vietnam in 2013 failed. Thus, we evaluated a series of relevant issues such as (a) why do some releases fail? (b) What affects the success of population replacement? And (c) Whether or not augmentation can block the dengue diseases in field trials. If not, how we can success be achieved? Models with and without augmentation, incorporating the effects of cytoplasmic incompatibility (CI) and fitness effects are proposed to describe the spread of Wolbachia in mosquito populations. Stability analysis revealed that backward bifurcations and multiple attractors may exist, which indicate that initial quantities of infected and uninfected mosquitoes, augmentation methods (timing, quantity, order and frequency) may affect the success of the strategies. The results show that successful population replacement will rely on selection of suitable strains of Wolbachia and careful design of augmentation methods.}, } @article {pmid26400107, year = {2016}, author = {Baldridge, GD and Li, YG and Witthuhn, BA and Higgins, L and Markowski, TW and Baldridge, AS and Fallon, AM}, title = {Mosaic composition of ribA and wspB genes flanking the virB8-D4 operon in the Wolbachia supergroup B-strain, wStr.}, journal = {Archives of microbiology}, volume = {198}, number = {1}, pages = {53-69}, pmid = {26400107}, issn = {1432-072X}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI 081322/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Base Sequence ; Genes, Bacterial/*genetics ; Operon/genetics ; Proteomics ; Wolbachia/*genetics ; }, abstract = {The obligate intracellular bacterium, Wolbachia pipientis (Rickettsiales), is a widespread, vertically transmitted endosymbiont of filarial nematodes and arthropods. In insects, Wolbachia modifies reproduction, and in mosquitoes, infection interferes with replication of arboviruses, bacteria and plasmodia. Development of Wolbachia as a tool to control pest insects will be facilitated by an understanding of molecular events that underlie genetic exchange between Wolbachia strains. Here, we used nucleotide sequence, transcriptional and proteomic analyses to evaluate expression levels and establish the mosaic nature of genes flanking the T4SS virB8-D4 operon from wStr, a supergroup B-strain from a planthopper (Hemiptera) that maintains a robust, persistent infection in an Aedes albopictus mosquito cell line. Based on protein abundance, ribA, which contains promoter elements at the 5'-end of the operon, is weakly expressed. The 3'-end of the operon encodes an intact wspB, which encodes an outer membrane protein and is co-transcribed with the vir genes. WspB and vir proteins are expressed at similar, above average abundance levels. In wStr, both ribA and wspB are mosaics of conserved sequence motifs from Wolbachia supergroup A- and B-strains, and wspB is nearly identical to its homolog from wCobU4-2, an A-strain from weevils (Coleoptera). We describe conserved repeated sequence elements that map within or near pseudogene lesions and transitions between A- and B-strain motifs. These studies contribute to ongoing efforts to explore interactions between Wolbachia and its host cell in an in vitro system.}, } @article {pmid26395600, year = {2015}, author = {Becerra, JX and Venable, GX and Saeidi, V}, title = {Erratum to: Wolbachia-Free Heteropterans Do Not Produce Defensive Chemicals or Alarm Pheromones.}, journal = {Journal of chemical ecology}, volume = {41}, number = {9}, pages = {869}, doi = {10.1007/s10886-015-0632-4}, pmid = {26395600}, issn = {1573-1561}, } @article {pmid26387062, year = {2015}, author = {Bergmann, M and Englert, T and Stuetzer, B and Hawley, JR and Lappin, MR and Hartmann, K}, title = {Prevalence of selected rickettsial infections in cats in Southern Germany.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {42}, number = {}, pages = {33-36}, doi = {10.1016/j.cimid.2015.08.003}, pmid = {26387062}, issn = {1878-1667}, mesh = {Anaplasma/genetics ; Anaplasmataceae Infections/epidemiology/microbiology/*veterinary ; Animals ; Cat Diseases/*epidemiology/microbiology/virology ; Cats ; Ehrlichia/genetics ; Germany/epidemiology ; Immunodeficiency Virus, Feline/genetics ; Lentivirus Infections/epidemiology/veterinary/virology ; Leukemia Virus, Feline/genetics ; Neorickettsia/genetics ; Polymerase Chain Reaction/methods ; Prevalence ; Retroviridae Infections/epidemiology/veterinary/virology ; Rickettsia Infections/epidemiology/prevention & control/*veterinary ; Thrombocytopenia/microbiology/veterinary ; Ticks/microbiology ; Tumor Virus Infections/epidemiology/veterinary ; Wolbachia/genetics ; }, abstract = {Prevalence of Anaplasma, Ehrlichia, Neorickettsia, and Wolbachia DNA in blood of 479 cats collected in different veterinary clinics in Southern Germany was determined using a previously published conventional PCR using 16S-23S intergenic spacer primers (5' CTG GGG ACT ACG GTC GCA AGA C 3' - forward; 5' CTC CAG TTT ATC ACT GGA AGT T 3' - reverse). Purified amplicons were sequenced to confirm genus and species. Associations between rickettsial infections, and feline immunodeficiency virus (FIV), as well as feline leukemia virus (FeLV) status were evaluated. Rickettsial prevalence was 0.4% (2/479; CI: 0.01-1.62%). In the two infected cats, Anaplasma phagocytophilum DNA was amplified. These cats came from different environment and had outdoor access. Both were ill with many of their problems likely related to other diseases. However, one cat had neutrophilia with left shift and the other thrombocytopenia potentially caused by their A. phagocytophilum infection. There was no significant difference in the FIV and FeLV status between A. phagocytophilum-negative and -positive cats. A. phagocytophilum can cause infection in cats in Southern Germany, and appropriate tick control is recommended.}, } @article {pmid26385192, year = {2015}, author = {Morrow, JL and Frommer, M and Royer, JE and Shearman, DC and Riegler, M}, title = {Wolbachia pseudogenes and low prevalence infections in tropical but not temperate Australian tephritid fruit flies: manifestations of lateral gene transfer and endosymbiont spillover?.}, journal = {BMC evolutionary biology}, volume = {15}, number = {}, pages = {202}, pmid = {26385192}, issn = {1471-2148}, mesh = {Animals ; Australia ; Cloning, Molecular ; Gene Transfer, Horizontal ; Haplotypes ; Multilocus Sequence Typing ; Phylogeny ; Pseudogenes ; Tephritidae/*classification/*microbiology/physiology ; Wolbachia/*genetics/*isolation & purification/physiology ; }, abstract = {BACKGROUND: Maternally inherited Wolbachia bacteria infect many insect species. They can also be transferred horizontally into uninfected host lineages. A Wolbachia spillover from an infected source population must occur prior to the establishment of heritable infections, but this spillover may be transient. In a previous study of tephritid fruit fly species of tropical Australia we detected a high incidence of identical Wolbachia strains in several species as well as Wolbachia pseudogenes. Here, we have investigated this further by analysing field specimens of 24 species collected along a 3,000 km climate gradient of eastern Australia.

RESULTS: Wolbachia sequences were detected in individuals of nine of the 24 (37 %) species. Seven (29 %) species displayed four distinct Wolbachia strains based on characterisation of full multi locus sequencing (MLST) profiles; the strains occurred as single and double infections in a small number of individuals (2-17 %). For the two remaining species all individuals had incomplete MLST profiles and Wolbachia pseudogenes that may be indicative of lateral gene transfer into host genomes. The detection of Wolbachia was restricted to northern Australia, including in five species that only occur in the tropics. Within the more widely distributed Bactrocera tryoni and Bactrocera neohumeralis, Wolbachia also only occurred in the north, and was not linked to any particular mitochondrial haplotypes.

CONCLUSIONS: The presence of Wolbachia pseudogenes at high prevalence in two species in absence of complete MLST profiles may represent footprints of historic infections that have been lost. The detection of identical low prevalence strains in a small number of individuals of seven species may question their role as reproductive manipulator and their vertical inheritance. Instead, the findings may be indicative of transient infections that result from spillover events from a yet unknown source. These spillover events appear to be restricted to northern Australia, without proliferation in host lineages further south. Our study highlights that tropical fruit fly communities contain Wolbachia pseudogenes and may be exposed to frequent horizontal Wolbachia transfer. It also emphasises that global estimates of Wolbachia frequencies may need to consider lateral gene transfer and Wolbachia spillover that may be regionally restricted, transient and not inherited.}, } @article {pmid26366194, year = {2015}, author = {Hoffmann, AA and Ross, PA and Rašić, G}, title = {Wolbachia strains for disease control: ecological and evolutionary considerations.}, journal = {Evolutionary applications}, volume = {8}, number = {8}, pages = {751-768}, pmid = {26366194}, issn = {1752-4571}, abstract = {Wolbachia are endosymbionts found in many insects with the potential to suppress vectorborne diseases, particularly through interfering with pathogen transmission. Wolbachia strains are highly variable in their effects on hosts, raising the issue of which attributes should be selected to ensure that the best strains are developed for disease control. This depends on their ability to suppress viral transmission, invade host populations, persist without loss of viral suppression and not interfere with other control strategies. The potential to achieve these objectives is likely to involve evolutionary constraints; viral suppression may be limited by the ability of infections to spread due to deleterious host fitness effects. However, there are exceptions to these patterns in both natural infections and in novel associations generated following interspecific transfer, suggesting that pathogen blockage, deleterious fitness effects and changes to reproductive biology might be at least partly decoupled to achieve ideal infection attributes. The stability of introduced Wolbachia and its effects on viral transmission remain unclear, but rapid evolutionary changes seem unlikely. Although deliberate transfers of Wolbachia across species remain particularly challenging, the availability of strains with desirable attributes should be expanded, taking advantage of the diversity available across thousands of strains in natural populations.}, } @article {pmid26362082, year = {2015}, author = {Brown, AM and Howe, DK and Wasala, SK and Peetz, AB and Zasada, IA and Denver, DR}, title = {Comparative Genomics of a Plant-Parasitic Nematode Endosymbiont Suggest a Role in Nutritional Symbiosis.}, journal = {Genome biology and evolution}, volume = {7}, number = {9}, pages = {2727-2746}, pmid = {26362082}, issn = {1759-6653}, mesh = {Animals ; Gene Ontology ; *Genome, Bacterial ; Genomics ; In Situ Hybridization, Fluorescence ; Nematoda/*microbiology ; Nutritional Physiological Phenomena/genetics ; Phylogeny ; Pseudogenes ; *Symbiosis ; Verrucomicrobia/classification/*genetics ; Vitis/parasitology ; }, abstract = {Bacterial mutualists can modulate the biochemical capacity of animals. Highly coevolved nutritional mutualists do this by synthesizing nutrients missing from the host's diet. Genomics tools have advanced the study of these partnerships. Here we examined the endosymbiont Xiphinematobacter (phylum Verrucomicrobia) from the dagger nematode Xiphinema americanum, a migratory ectoparasite of numerous crops that also vectors nepovirus. Previously, this endosymbiont was identified in the gut, ovaries, and eggs, but its role was unknown. We explored the potential role of this symbiont using fluorescence in situ hybridization, genome sequencing, and comparative functional genomics. We report the first genome of an intracellular Verrucomicrobium and the first exclusively intracellular non-Wolbachia nematode symbiont. Results revealed that Xiphinematobacter had a small 0.916-Mb genome with only 817 predicted proteins, resembling genomes of other mutualist endosymbionts. Compared with free-living relatives, conserved proteins were shorter on average, and there was large-scale loss of regulatory pathways. Despite massive gene loss, more genes were retained for biosynthesis of amino acids predicted to be essential to the host. Gene ontology enrichment tests showed enrichment for biosynthesis of arginine, histidine, and aromatic amino acids, as well as thiamine and coenzyme A, diverging from the profiles of relatives Akkermansia muciniphilia (in the human colon), Methylacidiphilum infernorum, and the mutualist Wolbachia from filarial nematodes. Together, these features and the location in the gut suggest that Xiphinematobacter functions as a nutritional mutualist, supplementing essential nutrients that are depleted in the nematode diet. This pattern points to evolutionary convergence with endosymbionts found in sap-feeding insects.}, } @article {pmid26357541, year = {2015}, author = {O'Shea, KL and Singh, ND}, title = {Tetracycline-exposed Drosophila melanogaster males produce fewer offspring but a relative excess of sons.}, journal = {Ecology and evolution}, volume = {5}, number = {15}, pages = {3130-3139}, pmid = {26357541}, issn = {2045-7758}, abstract = {A large diversity of species possesses endosymbionts; these endosymbionts can exhibit mutualistic, parasitic, and commensal relationships with their hosts. Previous work has consistently revealed that depleting endosymbiont titer with antibiotic treatment can significantly alter host fitness and function, particularly with respect to reproductive phenotypes. Although these findings are often interpreted as resulting from the breakdown of highly coevolved symbioses, it is possible that antibiotic treatment itself rather than endosymbiont removal contributes to the observed perturbations in reproductive phenotypes. Here, we investigate the effect of tetracycline treatment on sex ratio and male reproductive fitness using Drosophila melanogaster as a model system. Our results indicate that tetracycline-treated males produce a relative excess of sons. We also find that tetracycline treatment reduces the number of progeny produced by treated males but not treated females. These findings are independent of the effects of tetracycline on Wolbachia titer and implicate the antibiotic itself as mediating these changes. It is yet unclear whether the sex ratio shift and reduction in male reproductive fitness are direct or indirect consequences of tetracycline exposure, and more work is needed to determine the molecular mechanisms by which these disturbances in reproductive phenotypes arise. Our data highlight the importance of considering the potentially confounding effects of antibiotic treatment when investigating the effects of endosymbiont depletion on host phenotypes.}, } @article {pmid26354218, year = {2015}, author = {Noor Afizah, A and Roziah, A and Nazni, WA and Lee, HL}, title = {Detection of Wolbachia from field collected Aedes albopictus Skuse in Malaysia.}, journal = {The Indian journal of medical research}, volume = {142}, number = {2}, pages = {205-210}, pmid = {26354218}, issn = {0975-9174}, mesh = {Aedes/genetics/*microbiology ; Animals ; Dengue/diagnosis/*genetics/microbiology ; Female ; Humans ; Insect Vectors/genetics ; Malaysia ; Male ; *Phylogeny ; Wolbachia/genetics/*isolation & purification/virology ; }, abstract = {BACKGROUND & OBJECTIVES: Wolbachia-based vector control strategies have been proposed as a mean to augment the existing measures for controlling dengue vector. Prior to utilizing Wolbachia in novel vector control strategies, it is crucial to understand the Wolbachia-mosquito interactions. Many studies have only focused on the prevalence of Wolbachia in female Aedes albopictus with lack of attention on Wolbachia infection on the male Ae. albopictus which also affects the effective expression of Wolbachia induced- cytoplasmic incompatibility (CI). In this study, field surveys were conducted to screen for the infection status of Wolbachia in female and male Ae. albopictus from various habitats including housing areas, islands and seashore.

METHODS: Adult Ae. albopictus (n=104) were collected using human landing catches and hand aspirator. Standard ovitraps were also set in the selected areas for five days and the larvae were identified to species level. All the collected Ae. albopictus were screened for the presence of Wolbachia using multiplex polymerase chain reaction (PCR) and gene sequencing of Wolbachia surface protein (wsp) gene.

RESULTS: A 100 per cent positivity of Wolbachia infection was observed for individual Ae. albopictus screened. For pooled mosquitoes, 73 of the 76 pools (female) and 83 of the 87 pools (male) were positive with Wolbachia infection. The wsp gene sequence of the Wolbachia strain isolated from individual and pooled mosquitoes showed a 100 per cent homology with Wolbachia sp. of Ae. albopictus isolated from various geographical regions. Phylogenetic analysis based on wsp gene fragments showed that the isolates were clustered into groups A and B, respectively.

The results indicated that Wolbachia infection was widespread in Ae. albopictus population both in female and male Ae. albopictus. All the infected females were superinfected with both A and B strains while the infected males showed a combination of superinfection of A and B strains and single infection of B strain.}, } @article {pmid26350613, year = {2015}, author = {Lau, YL and Lee, WC and Xia, J and Zhang, G and Razali, R and Anwar, A and Fong, MY}, title = {Draft genome of Brugia pahangi: high similarity between B. pahangi and B. malayi.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {451}, pmid = {26350613}, issn = {1756-3305}, mesh = {Adenosine Triphosphatases/genetics/metabolism ; Aedes/parasitology ; Animals ; Bacterial Proteins/genetics/metabolism ; Brugia pahangi/*genetics ; Gene Expression Regulation/physiology ; Genome, Helminth/*genetics ; Helminth Proteins/genetics/metabolism ; Phylogeny ; Wolbachia/genetics/isolation & purification ; }, abstract = {BACKGROUND: Efforts to completely eradicate lymphatic filariasis from human population may be challenged by the emergence of Brugia pahangi as another zoonotic lymphatic filarial nematode. In this report, a genomic study was conducted to understand this species at molecular level.

METHODS: After blood meal on a B. pahangi-harbouring cat, the Aedes togoi mosquitoes were maintained to harvest infective third stage larvae, which were then injected into male Mongolian gerbils. Subsequently, adult B. pahangi were obtained from the infected gerbil for genomic DNA extraction. Sequencing and subsequently, construction of genomic libraries were performed. This was followed by genomic analyses and gene annotation analysis. By using archived protein sequences of B. malayi and a few other nematodes, clustering of gene orthologs and phylogenetics were conducted.

RESULTS: A total of 9687 coding genes were predicted. The genome of B. pahangi shared high similarity to that B. malayi genome, particularly genes annotated to fundamental processes. Nevertheless, 166 genes were considered to be unique to B. pahangi, which may be responsible for the distinct properties of B. pahangi as compared to other filarial nematodes. In addition, 803 genes were deduced to be derived from Wolbachia, an endosymbiont bacterium, with 44 of these genes intercalate into the nematode genome.

CONCLUSIONS: The reporting of B. pahangi draft genome contributes to genomic archive. Albeit with high similarity to B. malayi genome, the B. pahangi-unique genes found in this study may serve as new focus to study differences in virulence, vector selection and host adaptability among different Brugia spp.}, } @article {pmid26348356, year = {2015}, author = {Carrington, LB and Lipkowitz, JR and Hoffmann, AA and Turelli, M}, title = {Correction: A Re-Examination of Wolbachia-Induced Cytoplasmic Incompatibility in California Drosophila simulans.}, journal = {PloS one}, volume = {10}, number = {9}, pages = {e0138050}, pmid = {26348356}, issn = {1932-6203}, } @article {pmid26344089, year = {2015}, author = {Pontier, SM and Schweisguth, F}, title = {A Wolbachia-Sensitive Communication between Male and Female Pupae Controls Gamete Compatibility in Drosophila.}, journal = {Current biology : CB}, volume = {25}, number = {18}, pages = {2339-2348}, doi = {10.1016/j.cub.2015.07.052}, pmid = {26344089}, issn = {1879-0445}, support = {P40OD018537/OD/NIH HHS/United States ; }, mesh = {Animal Communication ; Animals ; Drosophila melanogaster/growth & development/*microbiology/*physiology ; Female ; Male ; Metamorphosis, Biological ; Olfactory Receptor Neurons/physiology ; Pupa/growth & development/microbiology/physiology ; Spermatozoa/growth & development/microbiology/physiology ; Testis/growth & development/microbiology ; Wolbachia/*physiology ; }, abstract = {Gamete compatibility is fundamental to sexual reproduction. Wolbachia are maternally inherited endosymbiotic bacteria that manipulate gamete compatibility in many arthropod species. In Drosophila, the fertilization of uninfected eggs by sperm from Wolbachia-infected males often results in early developmental arrest. This gamete incompatibility is called cytoplasmic incompatibility (CI). CI is highest in young males, suggesting that Wolbachia affect sperm properties during male development. Here, we show that Wolbachia modulate testis development. Unexpectedly, this effect was associated with Wolbachia infection in females, not males. This raised the possibility that females influenced testis development by communicating with males prior to adulthood. Using a combinatorial rearing protocol, we provide evidence for such a female-to-male communication during metamorphosis. This communication involves the perception of female pheromones by male olfactory receptors. We found that this communication determines the compatibility range of sperm. Wolbachia interfere with this female-to-male communication through changes in female pheromone production. Strikingly, restoring this communication partially suppressed CI in Wolbachia-infected males. We further identified a reciprocal male-to-female communication at metamorphosis that restricts the compatibility range of female gametes. Wolbachia also perturb this communication by feminizing male pheromone production. Thus, Wolbachia broaden the compatibility range of eggs, promoting thereby the reproductive success of Wolbachia-infected females. We conclude that pheromone communication between pupae regulates gamete compatibility and is sensitive to Wolbachia in Drosophila.}, } @article {pmid26336689, year = {2015}, author = {O'Neill, S}, title = {THE DENGUE STOPPER.}, journal = {Scientific American}, volume = {312}, number = {6}, pages = {72-77}, doi = {10.1038/scientificamerican0615-72}, pmid = {26336689}, issn = {0036-8733}, mesh = {Aedes/*microbiology ; Animals ; Dengue/*prevention & control/transmission ; Dengue Virus/growth & development ; Disease Transmission, Infectious/prevention & control ; Humans ; Insect Vectors/*microbiology ; Mosquito Control/*methods ; Wolbachia/*physiology ; }, } @article {pmid26312129, year = {2015}, author = {Solovyev, VI and Ilinsky, Y and Kosterin, OE}, title = {Genetic integrity of four species of Leptidea (Pieridae, Lepidoptera) as sampled in sympatry in West Siberia.}, journal = {Comparative cytogenetics}, volume = {9}, number = {3}, pages = {299-324}, pmid = {26312129}, issn = {1993-0771}, abstract = {In southern West Siberia, as many as four Leptidea Billberg, 1820 species are present sympatrically: Leptideaamurensis (Ménétriés, 1859), Leptideamorsei (Ménétriés, 1859), Leptideasinapis (Linnaeus, 1758) and Leptideajuvernica Williams, 1946. The two latter were recently recognised as nearly sibling species on morphological and molecular characters. Specimens intermediate as to their subtle diagnostic characters occurring in West Siberia and elsewhere were interpreted as resulted from limited introgression. This supposition was tested via populational morphological and molecular analysis of spring brood specimens of all the four species taken from a limited (4.5 × 0.2 km) area in the suburbs of Novosibirsk. The samples were analysed with respect to the genitalic morphology, external characters, three nuclear (CAD, H1 gene and ITS2) and one mitochondrial (COI) molecular markers, infection of the intracellular maternally inherited bacterial symbiont Wolbachia Hertig, 1836 and its wsp gene coding for a hypervariable surface protein. Interspecific variation of the nuclear CAD and ITS2 sequences and the mitochondrial COI gene in Leptideasinapis and Leptideajuvernica turned out concordant. The absence of molecular evidence of introgression suggests genetic integrity of these two species and allows their reliable identification by molecular characters. The genitalic (lengths of the saccus and valva) and external characters (wing pattern) of males overlap in Leptideasinapis and Leptideajuvernica, as identified by molecular markers and thus are not so helpful in actual species identification. Only the ductus bursae length showed no overlap and can be used for identification of females. The histone H1 gene appeared five times less variable over the four studied species than COI, and found to be identical in species Leptideasinapis and Leptideajuvernica. Wolbachia infection was found in all studied species. We identified three wsp variants of Wolbachia: 1) wsp-10 allele in Leptideaamurensis, Leptideasinapis, Leptideajuvernica; 2) a very similar wsp-687 allele in Leptideasinapis; and 3) wsp-688, highly divergent to the previous ones, in Leptideamorsei.}, } @article {pmid26309306, year = {2014}, author = {Andrews, ES and Fu, Y and Calvitti, M and Dobson, SL}, title = {Interspecific Transfer of a Wolbachia Infection Into Aedes albopictus (Diptera: Culicidae) Yields a Novel Phenotype Capable of Rescuing a Superinfection.}, journal = {Journal of medical entomology}, volume = {51}, number = {6}, pages = {1192-1198}, doi = {10.1603/ME14086}, pmid = {26309306}, issn = {0022-2585}, mesh = {Aedes/*microbiology ; Animals ; Female ; Male ; *Pest Control, Biological ; Phenotype ; Reproductive Isolation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally transmitted obligate bacteria that occur naturally in many arthropods. The phenotype observed in mosquitoes is known as cytoplasmic incompatibility (CI), which results in reduced or absent egg hatch in crosses between individuals with different infection types. Applied mosquito control strategies propose that by releasing individuals infected with a Wolbachia strain that differs from that in the natural host population, CI could be used to suppress or replace mosquito populations. Here, using tetracycline treatment and embryonic microinjection, Aedes albopictus (Skuse) was cleared of its natural Wolbachia infection and artificially infected with a Wolbachia strain originating from Aedes riversi Bohart & Ingram. Crossing experiments were carried out to determine whether CI could be observed between the artificially infected strain (UC), naturally infected (wild type), and uninfected strains of Ae. albopictus. Crosses between UC males and uninfected females resulted in no egg hatch, a classic unidirectional CI pattern. Crosses between the wild-type and UC strain also exhibited a unidirectional pattern of CI, demonstrating that the UC strain is compatible with both of the Wolbachia types that occur within Ae. albopictus and that wild-type Wolbachia infections are unable to fully rescue the UC Wolbachia type. Crosses between the UC strain and another artificially infected Ae. albopictus strain (ARwP), were bidirectionally incompatible, demonstrating that the UC strain is not compatible with all Wolbachia types. The CI patterns observed in this study were atypical and the opposite of that typically observed with superinfections.}, } @article {pmid26301521, year = {2015}, author = {Vale, PF and Jardine, MD}, title = {Sex-specific behavioural symptoms of viral gut infection and Wolbachia in Drosophila melanogaster.}, journal = {Journal of insect physiology}, volume = {82}, number = {}, pages = {28-32}, doi = {10.1016/j.jinsphys.2015.08.005}, pmid = {26301521}, issn = {1879-1611}, support = {095831//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Behavior, Animal ; Drosophila melanogaster/*microbiology/*virology ; Female ; Gastrointestinal Tract/virology ; Host-Pathogen Interactions ; Insect Viruses/*physiology ; Locomotion ; Male ; *Sex Factors ; Sleep ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {All organisms are infected with a range of symbionts spanning the spectrum of beneficial mutualists to detrimental parasites. The fruit fly Drosophila melanogaster is a good example, as both endosymbiotic Wolbachia, and pathogenic Drosophila C Virus (DCV) commonly infect it. While the pathophysiology and immune responses against both symbionts are the focus of intense study, the behavioural effects of these infections have received less attention. Here we report sex-specific behavioural responses to these infections in D. melanogaster. DCV infection caused increased sleep in female flies, but had no detectable effect in male flies. The presence of Wolbachia did not reduce this behavioural response to viral infection. We also found evidence for a sex-specific cost of Wolbachia, as male flies infected with the endosymbiont became more lethargic when awake. We discuss these behavioural symptoms as potentially adaptive sickness behaviours.}, } @article {pmid26291077, year = {2015}, author = {Flores, HA and Bubnell, JE and Aquadro, CF and Barbash, DA}, title = {The Drosophila bag of marbles Gene Interacts Genetically with Wolbachia and Shows Female-Specific Effects of Divergence.}, journal = {PLoS genetics}, volume = {11}, number = {8}, pages = {e1005453}, pmid = {26291077}, issn = {1553-7404}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; T32 GM007617/GM/NIGMS NIH HHS/United States ; GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila Proteins/*genetics/metabolism ; Drosophila melanogaster/genetics/*metabolism/microbiology ; Evolution, Molecular ; Female ; Gene Expression ; Genetic Complementation Test ; Host-Pathogen Interactions ; Infertility/genetics ; Male ; Ovary/metabolism/pathology ; Sex Characteristics ; Wolbachia/*physiology ; }, abstract = {Many reproductive proteins from diverse taxa evolve rapidly and adaptively. These proteins are typically involved in late stages of reproduction such as sperm development and fertilization, and are more often functional in males than females. Surprisingly, many germline stem cell (GSC) regulatory genes, which are essential for the earliest stages of reproduction, also evolve adaptively in Drosophila. One example is the bag of marbles (bam) gene, which is required for GSC differentiation and germline cyst development in females and for regulating mitotic divisions and entry to spermatocyte differentiation in males. Here we show that the extensive divergence of bam between Drosophila melanogaster and D. simulans affects bam function in females but has no apparent effect in males. We further find that infection with Wolbachia pipientis, an endosymbiotic bacterium that can affect host reproduction through various mechanisms, partially suppresses female sterility caused by bam mutations in D. melanogaster and interacts differentially with bam orthologs from D. melanogaster and D. simulans. We propose that the adaptive evolution of bam has been driven at least in part by the long-term interactions between Drosophila species and Wolbachia. More generally, we suggest that microbial infections of the germline may explain the unexpected pattern of evolution of several GSC regulatory genes.}, } @article {pmid26287231, year = {2015}, author = {Molloy, JC and Sinkins, SP}, title = {Wolbachia Do Not Induce Reactive Oxygen Species-Dependent Immune Pathway Activation in Aedes albopictus.}, journal = {Viruses}, volume = {7}, number = {8}, pages = {4624-4639}, pmid = {26287231}, issn = {1999-4915}, support = {095121//Wellcome Trust/United Kingdom ; BB/H016511/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Aedes/immunology/*microbiology ; Animals ; Arboviruses/immunology ; Cell Line ; Drosophila melanogaster/microbiology ; Female ; Gene Expression Profiling ; Immunity, Innate ; Reactive Oxygen Species/*metabolism ; Wolbachia/*immunology ; }, abstract = {Aedes albopictus is a major vector of dengue (DENV) and chikungunya (CHIKV) viruses, causing millions of infections annually. It naturally carries, at high frequency, the intracellular inherited bacterial endosymbiont Wolbachia strains wAlbA and wAlbB; transinfection with the higher-density Wolbachia strain wMel from Drosophila melanogaster led to transmission blocking of both arboviruses. The hypothesis that reactive oxygen species (ROS)-induced immune activation plays a role in arbovirus inhibition in this species was examined. In contrast to previous observations in Ae. aegypti, elevation of ROS levels was not observed in either cell lines or mosquito lines carrying the wild-type Wolbachia or higher-density Drosophila Wolbachia strains. There was also no upregulation of genes controlling innate immune pathways or with antioxidant/ROS-producing functions. These data suggest that ROS-mediated immune activation is not an important component of the viral transmission-blocking phenotype in this species.}, } @article {pmid26254486, year = {2015}, author = {Choi, JY and Bubnell, JE and Aquadro, CF}, title = {Population Genomics of Infectious and Integrated Wolbachia pipientis Genomes in Drosophila ananassae.}, journal = {Genome biology and evolution}, volume = {7}, number = {8}, pages = {2362-2382}, pmid = {26254486}, issn = {1759-6653}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; R01GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*microbiology ; *Evolution, Molecular ; Female ; *Genetic Variation ; *Genome, Bacterial ; Genome, Mitochondrial ; Genomics ; Heterozygote ; Polymorphism, Genetic ; Sequence Alignment ; Wolbachia/*genetics ; }, abstract = {Coevolution between Drosophila and its endosymbiont Wolbachia pipientis has many intriguing aspects. For example, Drosophila ananassae hosts two forms of W. pipientis genomes: One being the infectious bacterial genome and the other integrated into the host nuclear genome. Here, we characterize the infectious and integrated genomes of W. pipientis infecting D. ananassae (wAna), by genome sequencing 15 strains of D. ananassae that have either the infectious or integrated wAna genomes. Results indicate evolutionarily stable maternal transmission for the infectious wAna genome suggesting a relatively long-term coevolution with its host. In contrast, the integrated wAna genome showed pseudogene-like characteristics accumulating many variants that are predicted to have deleterious effects if present in an infectious bacterial genome. Phylogenomic analysis of sequence variation together with genotyping by polymerase chain reaction of large structural variations indicated several wAna variants among the eight infectious wAna genomes. In contrast, only a single wAna variant was found among the seven integrated wAna genomes examined in lines from Africa, south Asia, and south Pacific islands suggesting that the integration occurred once from a single infectious wAna genome and then spread geographically. Further analysis revealed that for all D. ananassae we examined with the integrated wAna genomes, the majority of the integrated wAna genomic regions is represented in at least two copies suggesting a double integration or single integration followed by an integrated genome duplication. The possible evolutionary mechanism underlying the widespread geographical presence of the duplicate integration of the wAna genome is an intriguing question remaining to be answered.}, } @article {pmid26252474, year = {2015}, author = {Zhang, D and Lees, RS and Xi, Z and Gilles, JR and Bourtzis, K}, title = {Combining the Sterile Insect Technique with Wolbachia-Based Approaches: II--A Safer Approach to Aedes albopictus Population Suppression Programmes, Designed to Minimize the Consequences of Inadvertent Female Release.}, journal = {PloS one}, volume = {10}, number = {8}, pages = {e0135194}, pmid = {26252474}, issn = {1932-6203}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; R01AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; *Cell Culture Techniques ; Crosses, Genetic ; Dose-Response Relationship, Radiation ; Female ; Fertility ; Host-Pathogen Interactions ; Infertility ; Insect Vectors/*microbiology ; Male ; Mosquito Control/*methods ; Ovary/radiation effects ; Polymerase Chain Reaction ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Due to the absence of a perfect method for mosquito sex separation, the combination of the sterile insect technique and the incompatible insect technique is now being considered as a potentially effective method to control Aedes albopictus. In this present study first we examine the minimum pupal irradiation dose required to induce complete sterility in Wolbachia triple-infected (HC), double-infected (GUA) and uninfected (GT) female Ae. albopictus. The HC line is a candidate for Ae. albopictus population suppression programmes, but due to the risk of population replacement which characterizes this triple infected line, the individuals to be released need to be additionally irradiated. After determining the minimum irradiation dose required for complete female sterility, we test whether sterilization is sufficient to prevent invasion of the triple infection from the HC females into double-infected (GUA) populations. Our results indicate that irradiated Ae. albopictus HC, GUA and GT strain females have decreased fecundity and egg hatch rate when irradiated, inversely proportional to the dose, and the complete sterilization of females can be acquired by pupal irradiation with doses above 28 Gy. PCR-based analysis of F1 and F2 progeny indicate that the irradiated HC females, cannot spread the new Wolbachia wPip strain into a small cage GUA population, released at a 1:5 ratio. Considering the above results, we conclude that irradiation can be used to reduce the risk of population replacement caused by an unintentional release of Wolbachia triple-infected Ae. albopictus HC strain females during male release for population suppression.}, } @article {pmid26244782, year = {2015}, author = {Duplouy, A and Couchoux, C and Hanski, I and van Nouhuys, S}, title = {Wolbachia Infection in a Natural Parasitoid Wasp Population.}, journal = {PloS one}, volume = {10}, number = {8}, pages = {e0134843}, pmid = {26244782}, issn = {1932-6203}, mesh = {Animals ; Electron Transport Complex IV/*genetics ; Female ; Fertility/genetics ; Finland ; Genetic Variation ; Geography ; Haplotypes ; Host-Pathogen Interactions/genetics ; Insect Proteins/*genetics ; Longevity/genetics ; Male ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Species Specificity ; Wasps/classification/*genetics/microbiology ; Wolbachia/classification/*genetics/physiology ; }, abstract = {The maternally transmitted bacterium Wolbachia pipientis is well known for spreading and persisting in insect populations through manipulation of the fitness of its host. Here, we identify three new Wolbachia pipientis strains, wHho, wHho2 and wHho3, infecting Hyposoter horticola, a specialist wasp parasitoid of the Glanville fritillary butterfly. The wHho strain (ST435) infects about 50% of the individuals in the Åland islands in Finland, with a different infection rate in the two mitochondrial (COI) haplotypes of the wasp. The vertical transmission rate of Wolbachia is imperfect, and lower in the haplotype with lower infection rate, suggesting a fitness trade-off. We found no association of the wHho infection with fecundity, longevity or dispersal ability of the parasitoid host. However, preliminary results convey spatial associations between Wolbachia infection, host mitochondrial haplotype and parasitism of H. horticola by its hyperparasitoid, Mesochorus cf. stigmaticus. We discuss the possibility that Wolbachia infection protects H. horticola against hyperparasitism.}, } @article {pmid26222703, year = {2015}, author = {Raquin, V and Valiente Moro, C and Saucereau, Y and Tran, FH and Potier, P and Mavingui, P}, title = {Correction: Native Wolbachia from Aedes albopictus Blocks Chikungunya Virus Infection In Cellulo.}, journal = {PloS one}, volume = {10}, number = {7}, pages = {e0134069}, pmid = {26222703}, issn = {1932-6203}, } @article {pmid26220534, year = {2015}, author = {Yuan, LL and Chen, X and Zong, Q and Zhao, T and Wang, JL and Zheng, Y and Zhang, M and Wang, Z and Brownlie, JC and Yang, F and Wang, YF}, title = {Quantitative Proteomic Analyses of Molecular Mechanisms Associated with Cytoplasmic Incompatibility in Drosophila melanogaster Induced by Wolbachia.}, journal = {Journal of proteome research}, volume = {14}, number = {9}, pages = {3835-3847}, doi = {10.1021/acs.jproteome.5b00191}, pmid = {26220534}, issn = {1535-3907}, mesh = {Animals ; Cytoplasm/*metabolism ; Drosophila melanogaster/*microbiology ; Male ; *Proteomics ; Real-Time Polymerase Chain Reaction ; Wolbachia/*pathogenicity ; }, abstract = {To investigate the molecular mechanisms of cytoplasmic incompatibility (CI) induced by Wolbachia bacteria in Drosophila melanogaster, we applied an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic assay to identify differentially expressed proteins extracted from spermathecae and seminal receptacles (SSR) of uninfected females mated with either 1-day-old Wolbachia-uninfected (1T) or infected males (1W) or 5-day-old infected males (5W). In total, 1317 proteins were quantified; 83 proteins were identified as having at least a 1.5-fold change in expression when 1W was compared with 1T. Differentially expressed proteins were related to metabolism, immunity, and reproduction. Wolbachia changed the expression of seminal fluid proteins (Sfps). Wolbachia may disrupt the abundance of proteins in SSR by affecting ubiquitin-proteasome-mediated proteolysis. Knocking down two Sfp genes (CG9334 and CG2668) in Wolbachia-free males resulted in significantly lower embryonic hatch rates with a phenotype of chromatin bridges. Wolbachia-infected females may rescue the hatch rates. This suggests that the changed expression of some Sfps may be one of the mechanisms of CI induced by Wolbachia. This study provides a panel of candidate proteins that may be involved in the interaction between Wolbachia and their insect hosts and, through future functional studies, may help to elucidate the underlying mechanisms of Wolbachia-induced CI.}, } @article {pmid26206894, year = {2015}, author = {Krawczyk, K and Szymańczyk, M and Obrępalska-Stęplowska, A}, title = {Prevalence of Endosymbionts in Polish Populations of Leptinotarsa decemlineata (Coleoptera: Chrysomelidae).}, journal = {Journal of insect science (Online)}, volume = {15}, number = {1}, pages = {}, pmid = {26206894}, issn = {1536-2442}, mesh = {Agriculture ; Animals ; Coleoptera/*microbiology ; DNA, Bacterial/genetics/metabolism ; Flavobacterium/genetics/metabolism/*physiology ; Molecular Sequence Data ; Phylogeny ; Poland ; RNA, Ribosomal, 23S/genetics/metabolism ; Sequence Analysis, DNA ; Solanum tuberosum/growth & development ; *Symbiosis ; }, abstract = {Colorado potato beetle (CPB, Leptinotarsa decemlineata Say) (Coleoptera: Chrysomelidae) is one of the most serious insect pest feeding on wild and cultivated Solanaceae plants. This pest poses a significant threat to potato crops. CPB originated from North America but has become widespread and has adapted in new localizations. Currently, it is reported in many countries worldwide. Endosymbiotic bacteria might have an influence on insect adaptation to new conditions. They are known to play a role in invasiveness of insect hosts and to facilitate colonization of new niches; however, information on endosymbionts of the CPB is very limited. In this study, we screened CPB populations collected from 20 evenly distributed locations in Poland for the presence of Arsenophonus, Cardinium, Wolbachia, and Flavobacterium. We found the presence of Flavobacterium in the studied insects. Little is known about CPB-endosymbionts interactions, thus this study may provide a reference for future studies in this subject.}, } @article {pmid26204449, year = {2015}, author = {Ritchie, SA and Townsend, M and Paton, CJ and Callahan, AG and Hoffmann, AA}, title = {Application of wMelPop Wolbachia Strain to Crash Local Populations of Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {7}, pages = {e0003930}, pmid = {26204449}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Female ; Host-Pathogen Interactions ; Infectious Disease Transmission, Vertical ; Male ; Ovum/microbiology ; Pest Control, Biological/*methods ; Population Dynamics ; Wolbachia/*classification/physiology ; }, abstract = {The endosymbiotic bacteria Wolbachia pipientis (wMel strain) has been successfully established in several populations of Aedes aegypti, the primary dengue vector. The virulent Wolbachia strain wMelPop is known to cause several pathological impacts (increased egg mortality, life shortening, etc.) reducing overall fitness in the mosquito Ae. aegypti. Increased egg mortality could substantially reduce egg banks in areas with a lengthy monsoonal dry season, and be employed to eliminate local populations. We tested this application under semi-field cage conditions. First, we determined that wMelPop infection significantly reduced the survival of desiccation-resistant eggs of the dengue vector Ae. aegypti, with shade and temperature having a significant impact; nearly all wMelPop-infected eggs failed to hatch after 6 and 10 weeks in summer and winter conditions, respectively. In laboratory selection experiments we found that egg desiccation resistance can be increased by selection, and that this effect of wMelPop infection is due to the nuclear background of the host rather than Wolbachia. We then conducted an invasion of wMelPop within a semi-field cage using sustained weekly releases of wMelPop infected mosquitoes, with fixation achieved after 9 weeks. The egg populations wMelPop infected and an uninfected control were then subjected to a simulated prolonged monsoonal dry season (2.5 months) before flooding to induce hatching. The wMelPop infected eggs suffered significantly greater mortality than the controls, with only 0.67% and 4.35% of respective infected and uninfected eggs held in 99% shade hatching after 80 days. These studies suggest that wMelPop could be used to locally eliminate populations of Ae. aegypti that are exposed to prolonged dry conditions, particularly if combined with vector control.}, } @article {pmid26189661, year = {2015}, author = {Ramírez-Puebla, ST and Servín-Garcidueñas, LE and Ormeño-Orrillo, E and Vera-Ponce de León, A and Rosenblueth, M and Delaye, L and Martínez, J and Martínez-Romero, E}, title = {Species in Wolbachia? Proposal for the designation of 'Candidatus Wolbachia bourtzisii', 'Candidatus Wolbachia onchocercicola', 'Candidatus Wolbachia blaxteri', 'Candidatus Wolbachia brugii', 'Candidatus Wolbachia taylori', 'Candidatus Wolbachia collembolicola' and 'Candidatus Wolbachia multihospitum' for the different species within Wolbachia supergroups.}, journal = {Systematic and applied microbiology}, volume = {38}, number = {6}, pages = {390-399}, doi = {10.1016/j.syapm.2015.05.005}, pmid = {26189661}, issn = {1618-0984}, mesh = {Animals ; Arthropods/microbiology ; Base Composition ; Computational Biology/*methods ; DNA, Bacterial/chemistry/*genetics ; *Genome, Bacterial ; Molecular Sequence Data ; Nematoda/microbiology ; Nucleic Acid Hybridization ; *Sequence Analysis, DNA ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia are highly extended bacterial endosymbionts that infect arthropods and filarial nematodes and produce contrasting phenotypes on their hosts. Wolbachia taxonomy has been understudied. Currently, Wolbachia strains are classified into phylogenetic supergroups. Here we applied phylogenomic analyses to study Wolbachia evolutionary relationships and examined metrics derived from their genome sequences such as average nucleotide identity (ANI), in silico DNA-DNA hybridization (DDH), G+C content, and synteny to shed light on the taxonomy of these bacteria. Draft genome sequences of strains wDacA and wDacB obtained from the carmine cochineal insect Dactylopius coccus were included. Although all analyses indicated that each Wolbachia supergroup represents a distinct evolutionary lineage, we found that some of the analyzed supergroups showed enough internal heterogeneity to be considered as assemblages of more than one species. Thus, supergroups would represent supraspecific groupings. Consequently, Wolbachia pipientis nomen species would apply only to strains of supergroup B and we propose the designation of 'Candidatus Wolbachia bourtzisii', 'Candidatus Wolbachia onchocercicola', 'Candidatus Wolbachia blaxterii', 'Candidatus Wolbachia brugii', 'Candidatus Wolbachia taylorii', 'Candidatus Wolbachia collembolicola' and 'Candidatus Wolbachia multihospitis' for other supergroups.}, } @article {pmid26180539, year = {2015}, author = {Ahmed, MZ and Araujo-Jnr, EV and Welch, JJ and Kawahara, AY}, title = {Wolbachia in butterflies and moths: geographic structure in infection frequency.}, journal = {Frontiers in zoology}, volume = {12}, number = {}, pages = {16}, pmid = {26180539}, issn = {1742-9994}, abstract = {INTRODUCTION: Butterflies and moths (Lepidoptera) constitute one of the most diverse insect orders, and play an important role in ecosystem function. However, little is known in terms of their bacterial communities. Wolbachia, perhaps the most common and widespread intracellular bacterium on Earth, can manipulate the physiology and reproduction of its hosts, and is transmitted vertically from mother to offspring, or sometimes horizontally between species. While its role in some hosts has been studied extensively, its incidence across Lepidoptera is poorly understood. A recent analysis using a beta-binomial model to infer the between-species distribution of prevalence estimated that approximately 40 % of arthropod species are infected with Wolbachia, but particular taxonomic groups and ecological niches seem to display substantially higher or lower incidences. In this study, we took an initial step and applied a similar, maximum likelihood approach to 300 species of Lepidoptera (7604 individuals from 660 populations) belonging to 17 families and 10 superfamilies, and sampled from 36 countries, representing all continents excluding Antarctica.

RESULTS: Approximately a quarter to a third of individuals appear to be infected with Wolbachia, and around 80 % of Lepidoptera species are infected at a non-negligible frequency. This incidence estimate is very high compared to arthropods in general. Wolbachia infection in Lepidoptera is shown to vary between families, but there is no evidence for closely related groups to show similar infection levels. True butterflies (Papilionoidea) are overrepresented in our data, however, our estimates show this group can be taken as a representative for the other major lepidopteran superfamilies. We also show substantial variation in infection level according to geography - closer locations tend to show similar infection levels. We further show that variation in geography is due to a latitudinal gradient in Wolbachia infection, with lower frequencies towards higher latitudes.

CONCLUSIONS: Our comprehensive survey of Wolbachia infection in Lepidoptera suggests that infection incidence is very high, and provides evidence that climate and geography are strong predictors of infection frequency.}, } @article {pmid26174827, year = {2015}, author = {Le, TA and Vi, TT and Nguyen, KL and Le, TH}, title = {A Rare Human Case of Dirofilaria repens Infection in the Subcutaneous Posterior Thorax with Molecular Identification.}, journal = {The Korean journal of parasitology}, volume = {53}, number = {3}, pages = {329-333}, pmid = {26174827}, issn = {1738-0006}, mesh = {Adult ; Animals ; Dirofilaria repens/classification/genetics/*isolation & purification/physiology ; Dirofilariasis/diagnosis/*parasitology ; Female ; Humans ; Phylogeny ; Thorax/*parasitology ; }, abstract = {The emergence of Dirofilarial infections in Asia including Vietnam is a clinically significant threat to the community. We here report a rare case of subcutaneous Dirofilaria repens infection on the posterior thoracic wall in a young woman presenting a painful, itchy, and palpable nodule. The adult worm was identified by mitochondrial cox1 and nuclear ITS-2 sequence determination. The diagnosis was additionally confirmed by 16S rRNA sequencing of the endosymbiont Wolbachia pipientis commonly co-existing with D. repens. This is a rare case of subcutaneous human infection on the posterior thoracic region caused by D. repens.}, } @article {pmid26172536, year = {2015}, author = {Fukui, T and Kawamoto, M and Shoji, K and Kiuchi, T and Sugano, S and Shimada, T and Suzuki, Y and Katsuma, S}, title = {The Endosymbiotic Bacterium Wolbachia Selectively Kills Male Hosts by Targeting the Masculinizing Gene.}, journal = {PLoS pathogens}, volume = {11}, number = {7}, pages = {e1005048}, pmid = {26172536}, issn = {1553-7374}, mesh = {Animals ; Female ; Male ; Molecular Sequence Data ; Moths/genetics/*parasitology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; *Wolbachia ; Zinc Fingers/genetics ; }, abstract = {Pathogens are known to manipulate the reproduction and development of their hosts for their own benefit. Wolbachia is an endosymbiotic bacterium that infects a wide range of insect species. Wolbachia is known as an example of a parasite that manipulates the sex of its host's progeny. Infection of Ostrinia moths by Wolbachia causes the production of all-female progeny, however, the mechanism of how Wolbachia accomplishes this male-specific killing is unknown. Here we show for the first time that Wolbachia targets the host masculinizing gene of Ostrinia to accomplish male-killing. We found that Wolbachia-infected O. furnacalis embryos do not express the male-specific splice variant of doublesex, a gene which acts at the downstream end of the sex differentiation cascade, throughout embryonic development. Transcriptome analysis revealed that Wolbachia infection markedly reduces the mRNA level of Masc, a gene that encodes a protein required for both masculinization and dosage compensation in the silkworm Bombyx mori. Detailed bioinformatic analysis also elucidated that dosage compensation of Z-linked genes fails in Wolbachia-infected O. furnacalis embryos, a phenomenon that is extremely similar to that observed in Masc mRNA-depleted male embryos of B. mori. Finally, injection of in vitro transcribed Masc cRNA into Wolbachia-infected embryos rescued male progeny. Our results show that Wolbachia-induced male-killing is caused by a failure of dosage compensation via repression of the host masculinizing gene. Our study also shows a novel strategy by which a pathogen hijacks the host sex determination cascade.}, } @article {pmid26172158, year = {2015}, author = {Webster, CL and Waldron, FM and Robertson, S and Crowson, D and Ferrari, G and Quintana, JF and Brouqui, JM and Bayne, EH and Longdon, B and Buck, AH and Lazzaro, BP and Akorli, J and Haddrill, PR and Obbard, DJ}, title = {The Discovery, Distribution, and Evolution of Viruses Associated with Drosophila melanogaster.}, journal = {PLoS biology}, volume = {13}, number = {7}, pages = {e1002210}, pmid = {26172158}, issn = {1545-7885}, support = {/WT_/Wellcome Trust/United Kingdom ; 095831/WT_/Wellcome Trust/United Kingdom ; WT085064/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Amino Acid Sequence ; Animals ; *Biological Evolution ; Conserved Sequence ; Drosophila melanogaster/*virology ; Drosophila simulans/virology ; Female ; Male ; Metagenomics ; Molecular Sequence Data ; RNA/analysis ; RNA Interference ; Viral Proteins/chemistry ; Wolbachia/isolation & purification ; }, abstract = {Drosophila melanogaster is a valuable invertebrate model for viral infection and antiviral immunity, and is a focus for studies of insect-virus coevolution. Here we use a metagenomic approach to identify more than 20 previously undetected RNA viruses and a DNA virus associated with wild D. melanogaster. These viruses not only include distant relatives of known insect pathogens but also novel groups of insect-infecting viruses. By sequencing virus-derived small RNAs, we show that the viruses represent active infections of Drosophila. We find that the RNA viruses differ in the number and properties of their small RNAs, and we detect both siRNAs and a novel miRNA from the DNA virus. Analysis of small RNAs also allows us to identify putative viral sequences that lack detectable sequence similarity to known viruses. By surveying >2,000 individually collected wild adult Drosophila we show that more than 30% of D. melanogaster carry a detectable virus, and more than 6% carry multiple viruses. However, despite a high prevalence of the Wolbachia endosymbiont--which is known to be protective against virus infections in Drosophila--we were unable to detect any relationship between the presence of Wolbachia and the presence of any virus. Using publicly available RNA-seq datasets, we show that the community of viruses in Drosophila laboratories is very different from that seen in the wild, but that some of the newly discovered viruses are nevertheless widespread in laboratory lines and are ubiquitous in cell culture. By sequencing viruses from individual wild-collected flies we show that some viruses are shared between D. melanogaster and D. simulans. Our results provide an essential evolutionary and ecological context for host-virus interaction in Drosophila, and the newly reported viral sequences will help develop D. melanogaster further as a model for molecular and evolutionary virus research.}, } @article {pmid26161659, year = {2015}, author = {Overholt, WA and Diaz, R and Rosskopf, E and Green, SJ and Overholt, WA}, title = {Deep Characterization of the Microbiomes of Calophya spp. (Hemiptera: Calophyidae) Gall-Inducing Psyllids Reveals the Absence of Plant Pathogenic Bacteria and Three Dominant Endosymbionts.}, journal = {PloS one}, volume = {10}, number = {7}, pages = {e0132248}, pmid = {26161659}, issn = {1932-6203}, mesh = {Animals ; Bacteria/*genetics ; Biodiversity ; Florida ; Hemiptera/*microbiology ; High-Throughput Nucleotide Sequencing/*methods ; Microbiota/*genetics ; Phylogeny ; Plant Tumors/*microbiology ; Symbiosis/*genetics ; }, abstract = {Bacteria associated with sap-feeding insect herbivores include not only symbionts that may increase their hosts' fitness but also harmful plant pathogens. Calophya spp. gall-inducing psyllids (Hemiptera: Calophyidae) are being investigated for their potential as biological control agents of the noxious weed, Brazilian peppertree (Schinus terebinthifolia), in Florida. Although there are no examples of plant pathogen transmission by members of the family Calophyidae, several insects in the superfamily Psylloidea are known to transmit pathogenic bacteria in the genera Candidatus Liberibacter and Candidatus Phytoplasma. To determine whether Calophya spp. harbor potentially harmful plant pathogenic bacteria, we sequenced small subunit (SSU) ribosomal RNA (rRNA) gene amplicons generated from individuals from four Calophya spp. populations: All microbial SSU gene sequences fell into the bacterial domain, with 98-99% belonging to the Proteobacteria. The Calophya microbiomes contained a relatively simple community, with 49-79 operational taxonomic units (OTUs; 97%) detected, and only 5-8 OTUs with greater than 1% abundance. Candidatus Carsonella showed the highest relative abundance, with OTUs from this candidate genus representing between 51-65% of all recovered sequences. The next most abundant clade observed was an unclassified Enterobacteriacae group closely related to bacteria from the genera Buchnera and Blochmannia that ranged from 20-31% in relative abundance. Wolbachia populations were the third most abundant group and represented 7-27% of the diversity in microbial OTUs. No SSU rRNA gene sequences from putative pathogenic bacteria from the genera Ca. Liberibacter or Ca. Phytoplasma were detected in the microbiomes of the four Calophya populations. The probability that infected psyllids were present in our colonies, but were not sampled, was extremley low (1.39 x 10(-10)). As far as we are aware, our study is the first to characterize the microbiome of a candidate biological control agent, and coupled with previous work demonstrating a high degree of host specificity and absence of plant viruses, suggests that releasing Calophya spp. in United States poses minimal risk to non-target plants.}, } @article {pmid26157152, year = {2015}, author = {Grankvist, A and Moore, ER and Svensson Stadler, L and Pekova, S and Bogdan, C and Geißdörfer, W and Grip-Lindén, J and Brandström, K and Marsal, J and Andréasson, K and Lewerin, C and Welinder-Olsson, C and Wennerås, C}, title = {Multilocus Sequence Analysis of Clinical "Candidatus Neoehrlichia mikurensis" Strains from Europe.}, journal = {Journal of clinical microbiology}, volume = {53}, number = {10}, pages = {3126-3132}, pmid = {26157152}, issn = {1098-660X}, mesh = {Aged ; Anaplasmataceae/*classification/*genetics/isolation & purification ; Anaplasmataceae Infections/epidemiology/*microbiology ; Cluster Analysis ; Czech Republic/epidemiology ; Female ; Genes, Essential ; *Genetic Variation ; *Genotype ; Germany/epidemiology ; Humans ; Male ; Middle Aged ; Molecular Epidemiology/methods ; Multilocus Sequence Typing/*methods ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sweden/epidemiology ; }, abstract = {"Candidatus Neoehrlichia mikurensis" is the tick-borne agent of neoehrlichiosis, an infectious disease that primarily affects immunocompromised patients. So far, the genetic variability of "Ca. Neoehrlichia" has been studied only by comparing 16S rRNA genes and groEL operon sequences. We describe the development and use of a multilocus sequence analysis (MLSA) protocol to characterize the genetic diversity of clinical "Ca. Neoehrlichia" strains in Europe and their relatedness to other species within the Anaplasmataceae family. Six genes were selected: ftsZ, clpB, gatB, lipA, groEL, and 16S rRNA. Each MLSA locus was amplified by real-time PCR, and the PCR products were sequenced. Phylogenetic trees of MLSA locus relatedness were constructed from aligned sequences. Blood samples from 12 patients with confirmed "Ca. Neoehrlichia" infection from Sweden (n = 9), the Czech Republic (n = 2), and Germany (n = 1) were analyzed with the MLSA protocol. Three of the Swedish strains exhibited identical lipA sequences, while the lipA sequences of the strains from the other nine patients were identical to each other. One of the Czech strains had one differing nucleotide in the clpB sequence from the sequences of the other 11 strains. All 12 strains had identical sequences for the genes 16S rRNA, ftsZ, gatB, and groEL. According to the MLSA, among the Anaplasmataceae, "Ca. Neoehrlichia" is most closely related to Ehrlichia ruminantium, less so to Anaplasma phagocytophilum, and least to Wolbachia endosymbionts. To conclude, three sequence types of infectious "Ca. Neoehrlichia" were identified: one in the west of Sweden, one in the Czech Republic, and one spread throughout Europe.}, } @article {pmid26150448, year = {2015}, author = {Nováková, E and Husník, F and Šochová, E and Hypša, V}, title = {Arsenophonus and Sodalis Symbionts in Louse Flies: an Analogy to the Wigglesworthia and Sodalis System in Tsetse Flies.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {18}, pages = {6189-6199}, pmid = {26150448}, issn = {1098-5336}, mesh = {Animals ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; Enterobacteriaceae/classification/genetics/*isolation & purification/physiology ; Genome, Bacterial ; Microscopy ; Molecular Sequence Data ; Phthiraptera/*microbiology/physiology ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology ; *Symbiosis ; Tsetse Flies/microbiology/physiology ; }, abstract = {Symbiosis between insects and bacteria result in a variety of arrangements, genomic modifications, and metabolic interconnections. Here, we present genomic, phylogenetic, and morphological characteristics of a symbiotic system associated with Melophagus ovinus, a member of the blood-feeding family Hippoboscidae. The system comprises four unrelated bacteria representing different stages in symbiosis evolution, from typical obligate mutualists inhabiting bacteriomes to freely associated commensals and parasites. Interestingly, the whole system provides a remarkable analogy to the association between Glossina and its symbiotic bacteria. In both, the symbiotic systems are composed of an obligate symbiont and two facultative intracellular associates, Sodalis and Wolbachia. In addition, extracellular Bartonella resides in the gut of Melophagus. However, the phylogenetic origins of the two obligate mutualist symbionts differ. In Glossina, the mutualistic Wigglesworthia appears to be a relatively isolated symbiotic lineage, whereas in Melophagus, the obligate symbiont originated within the widely distributed Arsenophonus cluster. Although phylogenetically distant, the two obligate symbionts display several remarkably similar traits (e.g., transmission via the host's "milk glands" or similar pattern of genome reduction). To obtain better insight into the biology and possible role of the M. ovinus obligate symbiont, "Candidatus Arsenophonus melophagi," we performed several comparisons of its gene content based on assignments of the Cluster of Orthologous Genes (COG). Using this criterion, we show that within a set of 44 primary and secondary symbionts, "Ca. Arsenophonus melophagi" is most similar to Wigglesworthia. On the other hand, these two bacteria also display interesting differences, such as absence of flagellar genes in Arsenophonus and their presence in Wigglesworthia. This finding implies that a flagellum is not essential for bacterial transmission via milk glands.}, } @article {pmid26150447, year = {2015}, author = {Mee, PT and Weeks, AR and Walker, PJ and Hoffmann, AA and Duchemin, JB}, title = {Detection of Low-Level Cardinium and Wolbachia Infections in Culicoides.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {18}, pages = {6177-6188}, pmid = {26150447}, issn = {1098-5336}, mesh = {Animals ; Australia ; Bacteroidetes/classification/genetics/*isolation & purification ; Ceratopogonidae/*microbiology ; Cluster Analysis ; DNA Gyrase/genetics ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Bacterial endosymbionts have been identified as potentially useful biological control agents for a range of invertebrate vectors of disease. Previous studies of Culicoides (Diptera: Ceratopogonidae) species using conventional PCR assays have provided evidence of Wolbachia (1/33) and Cardinium (8/33) infections. Here, we screened 20 species of Culicoides for Wolbachia and Cardinium, utilizing a combination of conventional PCR and more sensitive quantitative PCR (qPCR) assays. Low levels of Cardinium DNA were detected in females of all but one of the Culicoides species screened, and low levels of Wolbachia were detected in females of 9 of the 20 Culicoides species. Sequence analysis based on partial 16S rRNA gene and gyrB sequences identified "Candidatus Cardinium hertigii" from group C, which has previously been identified in Culicoides from Japan, Israel, and the United Kingdom. Wolbachia strains detected in this study showed 98 to 99% sequence identity to Wolbachia previously detected from Culicoides based on the 16S rRNA gene, whereas a strain with a novel wsp sequence was identified in Culicoides narrabeenensis. Cardinium isolates grouped to geographical regions independent of the host Culicoides species, suggesting possible geographical barriers to Cardinium movement. Screening also identified Asaia bacteria in Culicoides. These findings point to a diversity of low-level endosymbiont infections in Culicoides, providing candidates for further characterization and highlighting the widespread occurrence of these endosymbionts in this insect group.}, } @article {pmid26142572, year = {2015}, author = {Sugimoto, TN and Kayukawa, T and Matsuo, T and Tsuchida, T and Ishikawa, Y}, title = {A short, high-temperature treatment of host larvae to analyze Wolbachia-host interactions in the moth Ostrinia scapulalis.}, journal = {Journal of insect physiology}, volume = {81}, number = {}, pages = {48-51}, doi = {10.1016/j.jinsphys.2015.06.016}, pmid = {26142572}, issn = {1879-1611}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Female ; Host-Pathogen Interactions ; Hot Temperature ; Larva/drug effects/growth & development/microbiology ; Male ; Mosaicism ; Moths/drug effects/growth & development/*microbiology ; Sex Ratio ; Tetracycline/pharmacology ; Wolbachia/*physiology ; }, abstract = {Maternally inherited endosymbiotic bacteria of the genus Wolbachia cause various reproductive alterations in their hosts. Wolbachia induces male-specific death during embryonic and larval stages in the moth Ostrinia scapulalis. To investigate how the density of Wolbachia affects their performance in the host, we attempted to reduce its density using a short, high-temperature treatment of the host at the larval stage. Individuals cured of infection as well as sexual mosaics, which harbor Wolbachia, were obtained by this method in the next generation. The sex of uninfected offspring was exclusively male, similar to that of the offspring of larvae treated with antibiotics. A strong correlation was found between Wolbachia density in female moths and the sex ratio of their progeny. These results suggest that a short, high-temperature treatment at the larval stage reduced the density of Wolbachia in the adult stage, and, hence, inhibited interference with the host's development in the next generation. Since the direct effects of the heat treatment on Wolbachia were transient, this method may be useful for specifying the critical time for interference by Wolbachia in host development.}, } @article {pmid26139612, year = {2015}, author = {Choi, S and Shin, SK and Jeong, G and Yi, H}, title = {Wolbachia Sequence Typing in Butterflies Using Pyrosequencing.}, journal = {Journal of microbiology and biotechnology}, volume = {25}, number = {9}, pages = {1410-1416}, doi = {10.4014/jmb.1503.03097}, pmid = {26139612}, issn = {1738-8872}, mesh = {Animals ; Butterflies/*microbiology ; High-Throughput Nucleotide Sequencing/*methods ; Korea ; Multilocus Sequence Typing/*methods ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia is an obligate symbiotic bacteria that is ubiquitous in arthropods, with 25-70% of insect species estimated to be infected. Wolbachia species can interact with their insect hosts in a mutualistic or parasitic manner. Sequence types (ST) of Wolbachia are determined by multilocus sequence typing (MLST) of housekeeping genes. However, there are some limitations to MLST with respect to the generation of clone libraries and the Sanger sequencing method when a host is infected with multiple STs of Wolbachia. To assess the feasibility of massive parallel sequencing, also known as next-generation sequencing, we used pyrosequencing for sequence typing of Wolbachia in butterflies. We collected three species of butterflies (Eurema hecabe, Eurema laeta, and Tongeia fischeri) common to Korea and screened them for Wolbachia STs. We found that T. fischeri was infected with a single ST of Wolbachia, ST41. In contrast, E. hecabe and E. laeta were each infected with two STs of Wolbachia, ST41 and ST40. Our results clearly demonstrate that pyrosequencing-based MLST has a higher sensitivity than cloning and Sanger sequencing methods for the detection of minor alleles. Considering the high prevalence of infection with multiple Wolbachia STs, next-generation sequencing with improved analysis would assist with scaling up approaches to Wolbachia MLST.}, } @article {pmid26132467, year = {2015}, author = {Martinez, J and Ok, S and Smith, S and Snoeck, K and Day, JP and Jiggins, FM}, title = {Should Symbionts Be Nice or Selfish? Antiviral Effects of Wolbachia Are Costly but Reproductive Parasitism Is Not.}, journal = {PLoS pathogens}, volume = {11}, number = {7}, pages = {e1005021}, pmid = {26132467}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; WT094664MA/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila simulans/*microbiology ; Host-Parasite Interactions/*physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Symbiosis/*physiology ; *Wolbachia ; }, abstract = {Symbionts can have mutualistic effects that increase their host's fitness and/or parasitic effects that reduce it. Which of these strategies evolves depends in part on the balance of their costs and benefits to the symbiont. We have examined these questions in Wolbachia, a vertically transmitted endosymbiont of insects that can provide protection against viral infection and/or parasitically manipulate its hosts' reproduction. Across multiple symbiont strains we find that the parasitic phenotype of cytoplasmic incompatibility and antiviral protection are uncorrelated. Strong antiviral protection is associated with substantial reductions in other fitness-related traits, whereas no such trade-off was detected for cytoplasmic incompatibility. The reason for this difference is likely that antiviral protection requires high symbiont densities but cytoplasmic incompatibility does not. These results are important for the use of Wolbachia to block dengue virus transmission by mosquitoes, as natural selection to reduce these costs may lead to reduced symbiont density and the loss of antiviral protection.}, } @article {pmid26130134, year = {2015}, author = {Geary, JF and Lovato, R and Wanji, S and Guderian, R and O'Neill, M and Specht, S and Madrill, N and Geary, TG and Mackenzie, CD}, title = {A histochemical study of the Nras/let-60 activity in filarial nematodes.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {353}, pmid = {26130134}, issn = {1756-3305}, mesh = {Animals ; Bacterial Outer Membrane Proteins/analysis/metabolism ; Female ; Helminth Proteins/analysis/*metabolism ; Humans ; Immunohistochemistry ; Male ; Onchocerca/chemistry/*metabolism/microbiology ; Onchocerciasis/parasitology ; Wolbachia/chemistry/metabolism ; ras Proteins/analysis/*metabolism ; }, abstract = {BACKGROUND: Control and elimination of filarial pathogens is a central focus of major global health efforts directed at parasitic diseases of developing countries. Accomplishment of these goals would be markedly enhanced by the enhanced destruction of the adult stage of filariae. The identification of new, more quantitative biomarkers that correlate with mortality or chemotherapeutic damage to adult filariae, would greatly facilitate, for example, the development of new macrofilaricides.

METHODS: An immunocytochemical approach using an antibody against human Nras was used to identify and detect changes in the nematode homolog let-60 that is associated with cell growth and maintenance. Single Onchocerca volvulus nodules were removed from each of 13 patients treated with ivermectin (as part of a community-wide mass drug administration programme), and from each of 13 untreated individuals; these 26 nodules were stained with the anti-Nras antibody. The localization and degree of positivity of Nras/let-60 staining were assessed subjectively and compared between the two groups; the positivity of staining was also quantified, using image analysis, in a subgroup of these nodules. In addition, the specific morphological association between Nras/let-60 and the Wolbachia endosymbiont present in these parasites was also observed in 4 additional filarial species using an anti-Wolbachia surface protein (WSP) antibody under light and confocal microscopy.

RESULTS: Nras/let-60 is present in many structures within the adult female worms. A statistically significant decrease in the general staining intensity of Nras/let-60 was observed in adult female O. volvulus treated with ivermectin when compared with parasites from untreated patients. Nras/let-60 staining was frequently observed to be co-localized with WSP in O.volvulus, Brugia malayi, Litomosoides sigmodontis and Dirofilaria immitis. Nras/let60 is also present in Onchocerca ochengi.

CONCLUSION: Nras/let-60, as detected by immunocytochemical staining, is decreased in ivermectin-treated adult female O. volvulus relative to untreated control specimens, suggesting a suppressive effect of ivermectin on the overall biochemical activity of these parasites. Co-localization of Nras/let-60 and WSP suggests the possibility that the endosymbiont utilizes this nematode protein as part of a mutualistic relationship. Nras/let60 appears to be a useful biomarker for assessing the health of filariae.}, } @article {pmid26115104, year = {2015}, author = {Ye, YH and Carrasco, AM and Frentiu, FD and Chenoweth, SF and Beebe, NW and van den Hurk, AF and Simmons, CP and O'Neill, SL and McGraw, EA}, title = {Wolbachia Reduces the Transmission Potential of Dengue-Infected Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {6}, pages = {e0003894}, pmid = {26115104}, issn = {1935-2735}, support = {089276//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/microbiology/*virology ; Animals ; Dengue/*prevention & control/transmission ; Dengue Virus/*physiology ; Female ; Humans ; Insect Vectors/microbiology/*virology ; Male ; Pest Control, Biological/*methods ; Phenotype ; Saliva/virology ; Symbiosis ; Viral Load ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Dengue viruses (DENV) are the causative agents of dengue, the world's most prevalent arthropod-borne disease with around 40% of the world's population at risk of infection annually. Wolbachia pipientis, an obligate intracellular bacterium, is being developed as a biocontrol strategy against dengue because it limits replication of the virus in the mosquito. The Wolbachia strain wMel, which has been introduced into the mosquito vector, Aedes aegypti, has been shown to invade and spread to near fixation in field releases. Standard measures of Wolbachia's efficacy for blocking virus replication focus on the detection and quantification of virus in mosquito tissues. Examining the saliva provides a more accurate measure of transmission potential and can reveal the extrinsic incubation period (EIP), that is, the time it takes virus to arrive in the saliva following the consumption of DENV viremic blood. EIP is a key determinant of a mosquito's ability to transmit DENVs, as the earlier the virus appears in the saliva the more opportunities the mosquito will have to infect humans on subsequent bites.

We used a non-destructive assay to repeatedly quantify DENV in saliva from wMel-infected and Wolbachia-free wild-type control mosquitoes following the consumption of a DENV-infected blood meal. We show that wMel lengthens the EIP, reduces the frequency at which the virus is expectorated and decreases the dengue copy number in mosquito saliva as compared to wild-type mosquitoes. These observations can at least be partially explained by an overall reduction in saliva produced by wMel mosquitoes. More generally, we found that the concentration of DENV in a blood meal is a determinant of the length of EIP, saliva virus titer and mosquito survival.

CONCLUSIONS/SIGNIFICANCE: The saliva-based traits reported here offer more disease-relevant measures of Wolbachia's effects on the vector and the virus. The lengthening of EIP highlights another means, in addition to the reduction of infection frequencies and DENV titers in mosquitoes, by which Wolbachia should operate to reduce DENV transmission in the field.}, } @article {pmid26110714, year = {2015}, author = {Merkling, SH and van Rij, RP}, title = {Analysis of resistance and tolerance to virus infection in Drosophila.}, journal = {Nature protocols}, volume = {10}, number = {7}, pages = {1084-1097}, doi = {10.1038/nprot.2015.071}, pmid = {26110714}, issn = {1750-2799}, mesh = {Animals ; Drosophila melanogaster/genetics/microbiology/*virology ; Female ; Genes, Insect ; Genetic Variation ; Host-Pathogen Interactions ; Insect Viruses/genetics/*pathogenicity ; Male ; Symbiosis ; Wolbachia/physiology ; }, abstract = {Host defense to virus infection involves both resistance mechanisms that reduce viral burden and tolerance mechanisms that limit detrimental effects of infection. The fruit fly, Drosophila melanogaster, has emerged as a model for identifying and characterizing the genetic basis of resistance and tolerance. This protocol describes how to analyze host responses to virus infection in Drosophila, and it covers the preparation of virus stocks, experimental inoculation of flies and assessment of host survival and virus production, which are indicative of resistance or tolerance. It also provides guidance on how to account for recently identified confounding factors, including natural genetic variation in the pastrel locus and contamination of fly stocks with persistent viruses and the symbiotic bacterium Wolbachia. Our protocol aims to be accessible to newcomers to the field and, although optimized for virus research using Drosophila, some of the techniques could be adapted to other host organisms and/or other microbial pathogens. Preparation of fly stocks requires ∼1 month, virus stock preparation requires 17-20 d, virus injection and survival assays require 10-15 d and virus titration requires 14 d.}, } @article {pmid26102072, year = {2015}, author = {Hedtke, SM and Blitzer, EJ and Montgomery, GA and Danforth, BN}, title = {Introduction of Non-Native Pollinators Can Lead to Trans-Continental Movement of Bee-Associated Fungi.}, journal = {PloS one}, volume = {10}, number = {6}, pages = {e0130560}, pmid = {26102072}, issn = {1932-6203}, mesh = {Animal Migration ; Animals ; Aspergillus/classification/genetics/*isolation & purification/pathogenicity ; Bees/classification/*microbiology ; DNA, Bacterial/analysis ; DNA, Fungal/analysis ; Female ; Host-Pathogen Interactions ; Insecticides ; *Introduced Species ; Japan ; Likelihood Functions ; Male ; Malus ; Molecular Sequence Data ; Nesting Behavior ; New York ; Onygenales/classification/genetics/*isolation & purification/pathogenicity ; Paenibacillus/classification/genetics/isolation & purification/pathogenicity ; Phylogeny ; *Pollination ; Sequence Alignment ; Sequence Homology, Nucleic Acid ; Species Specificity ; }, abstract = {Bees are essential pollinators for many flowering plants, including agriculturally important crops such as apple. As geographic ranges of bees or their host plants change as a result of human activities, we need to identify pathogens that could be transmitted among newly sympatric species to evaluate and anticipate their effects on bee communities. We used PCR screening and DNA sequencing to evaluate exposure to potentially disease-causing microorganisms in a pollinator of apple, the horned mason bee (Osmia cornifrons). We did not detect microsporidia, Wolbachia, or trypanosomes, which are common pathogens of bees, in any of the hundreds of mason bees screened. We did detect both pathogenic and apathogenic (saprophytic) fungal species in the genus Ascosphaera (chalkbrood), an unidentified species of Aspergillus fungus, and a strain of bacteria in the genus Paenibacillus that is probably apathogenic. We detected pathogenic fungal strains in asymptomatic adult bees that therefore may be carriers of disease. We demonstrate that fungi from the genus Ascosphaera have been transported to North America along with the bee from its native range in Japan, and that O. cornifrons is exposed to fungi previously only identified from nests of other related bee species. Further study will be required to quantify pathogenicity and health effects of these different microbial species on O. cornifrons and on closely-related native North American mason bees that may now be exposed to novel pathogens. A global perspective is required for pathogen research as geographic ranges of insects and microorganisms shift due to intentional or accidental introductions.}, } @article {pmid26100152, year = {2015}, author = {Bah, GS and Tanya, VN and Makepeace, BL}, title = {Immunotherapy with mutated onchocystatin fails to enhance the efficacy of a sub-lethal oxytetracycline regimen against Onchocerca ochengi.}, journal = {Veterinary parasitology}, volume = {212}, number = {1-2}, pages = {25-34}, doi = {10.1016/j.vetpar.2015.06.005}, pmid = {26100152}, issn = {1873-2550}, mesh = {Animals ; Cameroon ; Cattle ; Cattle Diseases/drug therapy/*therapy ; Drug Therapy, Combination/standards/veterinary ; Female ; Helminth Proteins/administration & dosage/genetics/*immunology ; Immunoglobulin G/blood ; Immunotherapy/*veterinary ; Male ; Mutation ; Onchocerca/drug effects/physiology ; Oxytetracycline/pharmacology/*therapeutic use ; Random Allocation ; Treatment Outcome ; Vaccines/administration & dosage/chemistry ; }, abstract = {Human onchocerciasis (river blindness), caused by the filarial nematode Onchocerca volvulus, has been successfully controlled by a single drug, ivermectin, for over 25 years. Ivermectin prevents the disease symptoms of severe itching and visual impairment by killing the microfilarial stage, but does not eliminate the adult parasites, necessitating repeated annual treatments. Mass drug administration with ivermectin does not always break transmission in forest zones and is contraindicated in individuals heavily co-infected with Loa loa, while reports of reduced drug efficacy in Ghana and Cameroon may signal the development of resistance. An alternative treatment for onchocerciasis involves targeting the essential Wolbachia symbiont with tetracycline or its derivatives, which are adulticidal. However, implementation of antibiotic therapy has not occurred on a wide scale due to the prolonged treatment regimen required (several weeks). In the bovine Onchocerca ochengi system, it has been shown previously that prolonged oxytetracycline therapy increases eosinophil counts in intradermal nodules, which kill the adult worms by degranulating on their surface. Here, in an "immunochemotherapeutic" approach, we sought to enhance the efficacy of a short, sub-lethal antibiotic regimen against O. ochengi by prior immunotherapy targeting onchocystatin, an immunomodulatory protein located in the adult female worm cuticle. A key asparagine residue in onchocystatin was mutated to ablate immunomodulatory activity, which has been demonstrated previously to markedly improve the protective efficacy of this vaccine candidate when used as an immunoprophylactic. The immunochemotherapeutic regimen was compared with sub-lethal oxytetracycline therapy alone; onchocystatin immunotherapy alone; a gold-standard prolonged, intermittent oxytetracycline regimen; and no treatment (negative control) in naturally infected Cameroonian cattle. Readouts were collected over one year and comprised adult worm viability, dermal microfilarial density, anti-onchocystatin IgG in sera, and eosinophil counts in nodules. Only the gold-standard antibiotic regimen achieved significant killing of adult worms, a profound reduction in microfilarial load, and a sustained increase in local tissue eosinophilia. A small but statistically significant elevation in anti-onchocystatin IgG was observed for several weeks after immunisation in the immunotherapy-only group, but the antibody response in the immunochemotherapy group was more variable. At 12 weeks post-treatment, only a transient and non-significant increase in eosinophil counts was apparent in the immunochemotherapy group. We conclude that the addition of onchocystatin immunotherapy to a sub-lethal antibiotic regimen is insufficient to induce adulticidal activity, although with booster immunisations or the targeting of additional filarial immunomodulatory proteins, the efficacy of this strategy could be strengthened.}, } @article {pmid26086337, year = {2015}, author = {Jeffries, CL and Walker, T}, title = {The Potential Use of Wolbachia-Based Mosquito Biocontrol Strategies for Japanese Encephalitis.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {6}, pages = {e0003576}, pmid = {26086337}, issn = {1935-2735}, support = {/WT_/Wellcome Trust/United Kingdom ; 101285/WT_/Wellcome Trust/United Kingdom ; 101285/Z/13/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Cattle ; Culex/*microbiology ; Encephalitis Virus, Japanese/*physiology ; Encephalitis, Japanese/*prevention & control/transmission ; Humans ; Insect Vectors/*microbiology ; Pest Control, Biological/*methods ; Swine ; Wolbachia/*physiology ; Zoonoses/prevention & control ; }, abstract = {Japanese encephalitis virus (JEV) is a zoonotic pathogen transmitted by the infectious bite of Culex mosquitoes. The virus causes the development of the disease Japanese encephalitis (JE) in a small proportion of those infected, predominantly affecting children in eastern and southern Asia. Annual JE incidence estimates range from 50,000-175,000, with 25%-30% of cases resulting in mortality. It is estimated that 3 billion people live in countries in which JEV is endemic. The virus exists in an enzootic transmission cycle, with mosquitoes transmitting JEV between birds as reservoir hosts and pigs as amplifying hosts. Zoonotic infection occurs as a result of spillover events from the main transmission cycle. The reservoir avian hosts include cattle egrets, pond herons, and other species of water birds belonging to the family Ardeidae. Irrigated rice fields provide an ideal breeding ground for mosquitoes and attract migratory birds, maintaining the transmission of JEV. Although multiple vaccines have been developed for JEV, they are expensive and require multiple doses to maintain efficacy and immunity. As humans are a "dead-end" host for the virus, vaccination of the human population is unlikely to result in eradication. Therefore, vector control of the principal mosquito vector, Culex tritaeniorhynchus, represents a more promising strategy for reducing transmission. Current vector control strategies include intermittent irrigation of rice fields and space spraying of insecticides during outbreaks. However, Cx. Tritaeniorhynchus is subject to heavy exposure to pesticides in rice fields, and as a result, insecticide resistance has developed. In recent years, significant advancements have been made in the potential use of the bacterial endosymbiont Wolbachia for mosquito biocontrol. The successful transinfection of Wolbachia strains from Drosophila flies to Aedes (Stegomyia) mosquitoes has resulted in the generation of "dengue-refractory" mosquito lines. The successful establishment of Wolbachia in wild Aedes aegypti populations has recently been demonstrated, and open releases in dengue-endemic countries are ongoing. This review outlines the current control methods for JEV in addition to highlighting the potential use of Wolbachia-based biocontrol strategies to impact transmission. JEV and dengue virus are both members of the Flavivirus genus, and the successful establishment of Drosophila Wolbachia strains in Cx. Tritaeniorhynchus, as the principal vector of JEV, is predicted to significantly impact JEV transmission.}, } @article {pmid26075359, year = {2015}, author = {Wangwiwatsin, A and Protasio, AV}, title = {Genome watch. Adaptation: it's a bug's race.}, journal = {Nature reviews. Microbiology}, volume = {13}, number = {7}, pages = {401}, pmid = {26075359}, issn = {1740-1534}, mesh = {Animals ; Drosophila/*microbiology ; Gene Dosage ; Genome ; Nematoda/*microbiology ; Wolbachia/genetics/pathogenicity/*physiology ; }, abstract = {This month's Genome Watch highlights studies that investigate links between Wolbachia genotypes and their phenotypes, and explore how Wolbachia adapts to new host environments.}, } @article {pmid26070722, year = {2015}, author = {Becerra, JX and Venable, GX and Saeidi, V}, title = {Wolbachia-Free Heteropterans Do Not Produce Defensive Chemicals or Alarm Pheromones.}, journal = {Journal of chemical ecology}, volume = {41}, number = {7}, pages = {593-601}, pmid = {26070722}, issn = {1573-1561}, mesh = {Aldehydes/*metabolism ; Animals ; Anti-Bacterial Agents/pharmacology ; Heteroptera/*microbiology/*physiology ; Pheromones/*metabolism ; Phylogeny ; *Symbiosis ; Wolbachia/drug effects/genetics/isolation & purification/*physiology ; }, abstract = {The true bugs, or heteropterans, are known for their widespread production of anti-predator chemicals and alarm pheromones in scent glands, a derived trait that constitutes one of the defining characters of the suborder Heteroptera and a potential novel trait that contributed to their diversification. We investigated whether symbiotic bacteria could be involved in the formation of these chemicals using Thasus neocalifornicus, a coreid bug that produces semiochemicals frequently found in other bugs. Using DNA phylogenetic methodology and experiments using antibiotics coupled with molecular techniques, we identified Wolbachia as the microorganism infecting the scent glands of this bug. Decreasing the level of Wobachia infection using antibiotics was correlated with a diminution of heteropteran production of defensive compounds and alarm pheromones, suggesting that this symbiotic bacterium might be implicated in the formation of chemicals.}, } @article {pmid26051887, year = {2015}, author = {Lambrechts, L and Ferguson, NM and Harris, E and Holmes, EC and McGraw, EA and O'Neill, SL and Ooi, EE and Ritchie, SA and Ryan, PA and Scott, TW and Simmons, CP and Weaver, SC}, title = {Assessing the epidemiological effect of wolbachia for dengue control.}, journal = {The Lancet. Infectious diseases}, volume = {15}, number = {7}, pages = {862-866}, pmid = {26051887}, issn = {1474-4457}, support = {089276/WT_/Wellcome Trust/United Kingdom ; MR/K010174/1/MRC_/Medical Research Council/United Kingdom ; U01 GM110721/GM/NIGMS NIH HHS/United States ; }, mesh = {*Aedes ; Animals ; Dengue/*epidemiology/*prevention & control/transmission ; Dengue Virus/genetics ; Humans ; Insect Vectors/*microbiology ; Pest Control, Biological/*methods ; Randomized Controlled Trials as Topic ; Research Design ; Seroepidemiologic Studies ; *Wolbachia ; }, abstract = {Dengue viruses cause more human morbidity and mortality than any other arthropod-borne virus. Dengue prevention relies mainly on vector control; however, the failure of traditional methods has promoted the development of novel entomological approaches. Although use of the intracellular bacterium wolbachia to control mosquito populations was proposed 50 years ago, only in the past decade has its use as a potential agent of dengue control gained substantial interest. Here, we review evidence that supports a practical approach for dengue reduction through field release of wolbachia-infected mosquitoes and discuss the additional studies that have to be done before the strategy can be validated and implemented. A crucial next step is to assess the efficacy of wolbachia in reducing dengue virus transmission. We argue that a cluster randomised trial is at this time premature because choice of wolbachia strain for release and deployment strategies are still being optimised. We therefore present a pragmatic approach to acquiring preliminary evidence of efficacy through various complementary methods including a prospective cohort study, a geographical cluster investigation, virus phylogenetic analysis, virus surveillance in mosquitoes, and vector competence assays. This multipronged approach could provide valuable intermediate evidence of efficacy to justify a future cluster randomised trial.}, } @article {pmid26048932, year = {2015}, author = {Sapountzis, P and Zhukova, M and Hansen, LH and Sørensen, SJ and Schiøtt, M and Boomsma, JJ}, title = {Acromyrmex Leaf-Cutting Ants Have Simple Gut Microbiota with Nitrogen-Fixing Potential.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {16}, pages = {5527-5537}, pmid = {26048932}, issn = {1098-5336}, support = {323085/ERC_/European Research Council/International ; }, mesh = {Alphaproteobacteria/genetics/*isolation & purification/physiology ; Animals ; Ants/*microbiology ; Entomoplasmatales/*isolation & purification/physiology ; *Gastrointestinal Microbiome ; *Nitrogen Fixation ; Oxidoreductases/genetics ; Symbiosis ; Wolbachia/*isolation & purification/physiology ; }, abstract = {Ants and termites have independently evolved obligate fungus-farming mutualisms, but their gardening procedures are fundamentally different, as the termites predigest their plant substrate whereas the ants deposit it directly on the fungus garden. Fungus-growing termites retained diverse gut microbiota, but bacterial gut communities in fungus-growing leaf-cutting ants have not been investigated, so it is unknown whether and how they are specialized on an exclusively fungal diet. Here we characterized the gut bacterial community of Panamanian Acromyrmex species, which are dominated by only four bacterial taxa: Wolbachia, Rhizobiales, and two Entomoplasmatales taxa. We show that the Entomoplasmatales can be both intracellular and extracellular across different gut tissues, Wolbachia is mainly but not exclusively intracellular, and the Rhizobiales species is strictly extracellular and confined to the gut lumen, where it forms biofilms along the hindgut cuticle supported by an adhesive matrix of polysaccharides. Tetracycline diets eliminated the Entomoplasmatales symbionts but hardly affected Wolbachia and only moderately reduced the Rhizobiales, suggesting that the latter are protected by the biofilm matrix. We show that the Rhizobiales symbiont produces bacterial NifH proteins that have been associated with the fixation of nitrogen, suggesting that these compartmentalized hindgut symbionts alleviate nutritional constraints emanating from an exclusive fungus garden diet reared on a substrate of leaves.}, } @article {pmid26047139, year = {2015}, author = {Badawi, M and Grève, P and Cordaux, R}, title = {Feminization of the Isopod Cylisticus convexus after Transinfection of the wVulC Wolbachia Strain of Armadillidium vulgare.}, journal = {PloS one}, volume = {10}, number = {6}, pages = {e0128660}, pmid = {26047139}, issn = {1932-6203}, mesh = {Animals ; Comparative Genomic Hybridization ; Female ; *Feminization ; Gonads/anatomy & histology/metabolism/microbiology ; Isopoda/*microbiology/physiology ; Male ; Microscopy ; Phenotype ; Wolbachia/*physiology ; }, abstract = {Reproductive parasites such as Wolbachia are able to manipulate the reproduction of their hosts by inducing parthenogenesis, male-killing, cytoplasmic incompatibility or feminization of genetic males. Despite extensive studies, no underlying molecular mechanism has been described to date. The goal of this study was to establish a system with a single Wolbachia strain that feminizes two different isopod species to enable comparative analyses aimed at elucidating the genetic basis of feminization. It was previously suggested that Wolbachia wVulC, which naturally induces feminization in Armadillidium vulgare, induces the development of female secondary sexual characters in transinfected Cylisticus convexus adult males. However, this does not demonstrate that wVulC induces feminization in C. convexus since feminization is the conversion of genetic males into functional females that occurs during development. Nevertheless, it suggests that C. convexus may represent a feminization model suitable for further development. Knowledge about C. convexus sexual differentiation is also essential for comparative analyses, as feminization is thought to take place just before or during sexual differentiation. Consequently, we first described gonad morphological differentiation of C. convexus and compared it with that of A. vulgare. Then, wVulC was injected into male and female C. convexus adult individuals. The feminizing effect was demonstrated by the combined appearance of female secondary sexual characters in transinfected adult males, as well as the presence of intersexes and female biases in progenies in which wVulC was vertically transmitted from transinfected mothers. The establishment of a new model of feminization of a Wolbachia strain in a heterologous host constitutes a useful tool towards the understanding of the molecular mechanism of feminization.}, } @article {pmid26037532, year = {2015}, author = {Baldacchino, F and Caputo, B and Chandre, F and Drago, A and della Torre, A and Montarsi, F and Rizzoli, A}, title = {Control methods against invasive Aedes mosquitoes in Europe: a review.}, journal = {Pest management science}, volume = {71}, number = {11}, pages = {1471-1485}, doi = {10.1002/ps.4044}, pmid = {26037532}, issn = {1526-4998}, mesh = {*Aedes ; Animals ; Animals, Genetically Modified ; Ecosystem ; Environment ; Europe ; Insect Vectors ; Insecticides ; Introduced Species ; Mosquito Control/*methods ; Pest Control, Biological/methods ; }, abstract = {Five species of invasive Aedes mosquitoes have recently become established in Europe: Ae. albopictus, Ae. aegypti, Ae. japonicus japonicus, Ae. koreicus and Ae. atropalpus. These mosquitoes are a serious nuisance for people and are also competent vectors for several exotic pathogens such as dengue and chikungunya viruses. As they are a growing public health concern, methods to control these mosquitoes need to be implemented to reduce their biting and their potential for disease transmission. There is a crucial need to evaluate methods as part of an integrated invasive mosquito species control strategy in different European countries, taking into account local Aedes infestations and European regulations. This review presents the control methods available or in development against invasive Aedes mosquitoes, with a particular focus on those that can be implemented in Europe. These control methods are divided into five categories: environmental (source reduction), mechanical (trapping), biological (e.g. copepods, Bacillus thuringiensis var. israelensis, Wolbachia), chemical (insect growth regulators, pyrethroids) and genetic (sterile insect technique and genetically modified mosquitoes). We discuss the effectiveness, ecological impact, sustainability and stage of development of each control method.}, } @article {pmid26019119, year = {2015}, author = {Fallon, AM}, title = {Effects of mimosine on Wolbachia in mosquito cells: cell cycle suppression reduces bacterial abundance.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {51}, number = {9}, pages = {958-963}, pmid = {26019119}, issn = {1543-706X}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI 081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*cytology/*drug effects/microbiology ; Animals ; Cell Cycle/drug effects ; Cell Line/drug effects/microbiology ; Dose-Response Relationship, Drug ; Mimosine/*pharmacology ; Pheromones/pharmacology ; Wolbachia/*drug effects/growth & development/pathogenicity ; }, abstract = {The plant allelochemical L-mimosine (β-[N-(3-hydroxy-4-pyridone)]-α-aminopropionic acid; leucenol) resembles the nonessential amino acid, tyrosine. Because the obligate intracellular alphaproteobacterium, Wolbachia pipientis, metabolizes amino acids derived from host cells, the effects of mimosine on infected and uninfected mosquito cells were investigated. The EC50 for mimosine was 6-7 μM with Aedes albopictus C7-10 and C/wStr cell lines, and was not influenced by infection status. Mosquito cells responded to concentrations of mimosine substantially lower than those used to synchronize the mammalian cell cycle; at concentrations of 30-35 μM, mimosine reversibly arrested the mosquito cell cycle at the G1/S boundary and inhibited growth of Wolbachia strain wStr. Although lower concentrations of mimosine slightly increased wStr abundance, concentrations that suppressed the cell cycle reduced Wolbachia levels.}, } @article {pmid26004817, year = {2015}, author = {Chaisiri, K and McGarry, JW and Morand, S and Makepeace, BL}, title = {Symbiosis in an overlooked microcosm: a systematic review of the bacterial flora of mites.}, journal = {Parasitology}, volume = {142}, number = {9}, pages = {1152-1162}, doi = {10.1017/S0031182015000530}, pmid = {26004817}, issn = {1469-8161}, mesh = {Animals ; Bacteria/classification/*isolation & purification ; Bacterial Physiological Phenomena ; Mites/*microbiology ; Symbiosis ; }, abstract = {A dataset of bacterial diversity found in mites was compiled from 193 publications (from 1964 to January 2015). A total of 143 mite species belonging to the 3 orders (Mesostigmata, Sarcoptiformes and Trombidiformes) were recorded and found to be associated with approximately 150 bacteria species (in 85 genera, 51 families, 25 orders and 7 phyla). From the literature, the intracellular symbiont Cardinium, the scrub typhus agent Orientia, and Wolbachia (the most prevalent symbiont of arthropods) were the dominant mite-associated bacteria, with approximately 30 mite species infected each. Moreover, a number of bacteria of medical and veterinary importance were also reported from mites, including species from the genera Rickettsia, Anaplasma, Bartonella, Francisella, Coxiella, Borrelia, Salmonella, Erysipelothrix and Serratia. Significant differences in bacterial infection patterns among mite taxa were identified. These data will not only be useful for raising awareness of the potential for mites to transmit disease, but also enable a deeper understanding of the relationship of symbionts with their arthropod hosts, and may facilitate the development of intervention tools for disease vector control. This review provides a comprehensive overview of mite-associated bacteria and is a valuable reference database for future research on mites of agricultural, veterinary and/or medical importance.}, } @article {pmid25991706, year = {2015}, author = {Konecka, E and Olszanowski, Z}, title = {A screen of maternally inherited microbial endosymbionts in oribatid mites (Acari: Oribatida).}, journal = {Microbiology (Reading, England)}, volume = {161}, number = {8}, pages = {1561-1571}, doi = {10.1099/mic.0.000114}, pmid = {25991706}, issn = {1465-2080}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; *Bacterial Physiological Phenomena ; Bacterial Proteins/genetics/metabolism ; Female ; Host Specificity ; Male ; Mites/classification/*microbiology/physiology ; Phylogeny ; Species Specificity ; *Symbiosis ; Wolbachia/classification/genetics/isolation & purification/physiology ; }, abstract = {We determined the distribution of microbial endosymbionts as possible agents of parthenogenesis in Oribatida. We screened mites from 20 species of 14 families suspected to be parthenogenetic from the absence or rarity of males. Our research included parthenogenesis-inducing bacteria Wolbachia spp., Cardinium spp., Rickettsia spp., and additionally Arsenophonus, Spiroplasma and microsporidia that can also manipulate host reproduction. We detected the endosymbionts by PCR-based methods and transmission electron microscopy (TEM) observation of fixed and stained preparations of host cells. We detected Wolbachia only in one Oribatida species, Oppiella nova, by identifying Wolbachia genes using PCR. TEM observations confirmed infection by the endosymbiont in O. nova and its lack in other Oribatida species. Sequence analysis of hcpA and fbpA genes showed that the Wolbachia strain from O. nova was different from strains characterized in some insects, crustaceans (Isopoda), mites (Tetranychidae), springtails (Hexapoda) and roundworms (Nematoda). The analysis strongly suggested that the Wolbachia sp. strain found in O. nova did not belong to supergroups A, B, C, D, E, F, H or M. We found that the sequences of Wolbachia from O. nova were clearly distantly related to sequences from the bacteria of the other supergroups. This observation makes O. nova a unique Wolbachia host in terms of the distinction of the strain. The role of these micro-organisms in O. nova remains unknown and is an issue to investigate.}, } @article {pmid25989164, year = {2015}, author = {Otabil, KB and Tenkorang, SB}, title = {Filarial hydrocele: a neglected condition of a neglected tropical disease.}, journal = {Journal of infection in developing countries}, volume = {9}, number = {5}, pages = {456-462}, doi = {10.3855/jidc.5346}, pmid = {25989164}, issn = {1972-2680}, mesh = {Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Elephantiasis, Filarial/*complications ; Humans ; Male ; Neglected Diseases/*drug therapy/*pathology ; Testicular Hydrocele/*drug therapy/*pathology ; Tropical Climate ; }, abstract = {Filarial hydrocele is the most common chronic manifestation of lymphatic filariasis (LF) and poses a major public health burden to several filarial endemic countries. This review highlights the socio-economic impact of the disease, the role of the immune system in hydrocele development, current diagnostic approaches, and the control and management of filarial hydrocele. In the quest to facilitate the global effort to eliminate filarial hydrocele as a neglected tropical disease, a more comprehensive understanding of the mechanisms underlying the pathogenesis and development of the condition is important. In general, success has been achieved using annual treatment with ivermectin, but much remains to be done, particularly with late-stage infected individuals where surgery remains the only option. Studies have successfully demonstrated that inhibition of embryogenesis in adult female worms occurs after weeks of tetracycline treatment. Even more intriguing was the observation that the Wolbachia endosymbionts potently induce proinflammatory cytokines such as tumor necrosis factors (TNFs) and vascular endothelial growth factors (VEGFs), which are crucial for the development of filarial hydrocele. Furthermore, reports from human studies show that doxycycline treatment significantly ameliorates filarial hydrocele and markedly reverses early-stage filarial hydrocele. However, with the enormous challenges that face LF elimination such as global funding, logistics, civil wars, and drug resistance, a more relentless and collective approach from local governments as well as other stakeholders is needed to accelerate the fight against filarial hydrocele if the goal to eliminate it by 2020 is be to achieved.}, } @article {pmid25983231, year = {2015}, author = {Mbewe, NJ and Mweempwa, C and Guya, S and Wamwiri, FN}, title = {Microbiome frequency and their association with trypanosome infection in male Glossina morsitans centralis of Western Zambia.}, journal = {Veterinary parasitology}, volume = {211}, number = {1-2}, pages = {93-98}, doi = {10.1016/j.vetpar.2015.04.027}, pmid = {25983231}, issn = {1873-2550}, support = {D43 TW007391/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; Enterobacteriaceae/genetics/*isolation & purification ; Female ; Male ; *Microbiota ; Symbiosis ; Trypanosoma/*physiology ; Trypanosomiasis/*prevention & control ; Tsetse Flies/*microbiology/parasitology ; Wolbachia/genetics/*isolation & purification ; Zambia/epidemiology ; }, abstract = {Tsetse flies (Diptera: Glossinidae) are considered primary cyclical vectors that transmit pathogenic trypanosomes in Africa. They harbour a variety of microbes including Wolbachia, Sodalis and the salivary gland hypertrophy virus (SGHV) which are all vertically transmitted. Knowledge on tsetse microbiome and their interactions may identify novel strategies for tsetse fly and trypanosomiasis control. Area-wide application of such strategies requires an understanding of the natural microbiome frequency in the different species and subspecies of Glossina in their geographical populations. Consequently, this study determined the prevalence of Sodalis, Wolbachia, SGHV and trypanosome infections in Glossina morsitanscentralis from two sites of Western Zambia. We also explored possible associations of the microbes with trypanosome infections. Male G. morsitanscentralis samples were collected from two sites (Lyoni and Lusinina) in Western Zambia. The age structure of the flies at each site was determined using the wing fray method. DNA was extracted from the samples and analyzed for Wolbachia, Sodalis, SGHV and trypanosome presence using PCR. Associations and measures of associations between trypanosome infection and microbes in the fly were determined. The flies from the two locations (Lusinina, n=45 and Lyoni, n=24) had a similar age structure with their median fray category not being significantly different (p=0.698). The overall prevalence of Wolbachia was 72.5% (95% CI: 61.6-83.3%), Sodalis was 15.9% (95% CI: 7.1-24.8%), SGHV was 31.9% (95% CI: 20.6-43.2%) and Trypanosoma species was 23.2% (95% CI: 13-33.4%). The prevalence of Wolbachia was significantly higher in Lusinina than Lyoni (p=0.000). However this was not the case for Sodalis, SGHV and Trypanosoma species. Despite the low number of flies that were positive for both trypanosome and Sodalis (6; 8.7%), a statistically significant association (p=0.013; AOR 6.2; 95% CI: 1.5-25.8) was observed in G. morsitanscentralis. The study showed that the prevalence of microbiota may vary within the same species of the tsetse depending on the geographical location as was the case of Wolbachia. Further it showed that infection with Sodalis could affect vector competence. The study concludes that Sodalis could be an ideal candidate for symbiont-mediated trypanosomiasis control interventions in G. morsitanscentralis.}, } @article {pmid25981386, year = {2015}, author = {Rossi, P and Ricci, I and Cappelli, A and Damiani, C and Ulissi, U and Mancini, MV and Valzano, M and Capone, A and Epis, S and Crotti, E and Chouaia, B and Scuppa, P and Joshi, D and Xi, Z and Mandrioli, M and Sacchi, L and O'Neill, SL and Favia, G}, title = {Mutual exclusion of Asaia and Wolbachia in the reproductive organs of mosquito vectors.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {278}, pmid = {25981386}, issn = {1756-3305}, mesh = {Aedes/*microbiology ; Alphaproteobacteria/*isolation & purification ; Animals ; Anopheles/*microbiology ; Female ; Gastrointestinal Tract/microbiology ; Gonads/*microbiology ; Male ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia is a group of intracellular maternally inherited bacteria infecting a high number of arthropod species. Their presence in different mosquito species has been largely described, but Aedes aegypti, the main vector of Dengue virus, has never been found naturally infected by Wolbachia. Similarly, malaria vectors and other anophelines are normally negative to Wolbachia, with the exception of an African population where these bacteria have recently been detected. Asaia is an acetic acid bacterium stably associated with several mosquito species, found as a dominant microorganism of the mosquito microbiota. Asaia has been described in gut, salivary glands and in reproductive organs of adult mosquitoes in Ae. aegypti and in anophelines. It has recently been shown that Asaia may impede vertical transmission of Wolbachia in Anopheles mosquitoes. Here we present an experimental study, aimed at determining whether there is a negative interference between Asaia and Wolbachia, for the gonad niche in mosquitoes.

METHODS: Different methods (PCR and qPCR, monoclonal antibody staining and FISH) have been used to address the question of the co-localization and the relative presence/abundance of the two symbionts. PCR and qPCR were performed to qualitatively and quantitatively verify the distribution of Asaia and Wolbachia in different mosquito species/organs. Monoclonal antibody staining and FISH were performed to localize the symbionts in different mosquito species.

RESULTS: Here we provide evidence that, in Anopheles and in other mosquitoes, there is a reciprocal negative interference between Asaia and Wolbachia symbionts, in terms of the colonization of the gonads. In particular, we have shown that in some mosquito species the presence of one of the symbionts prevented the establishment of the second, while in other systems the symbionts were co-localized, although at reduced densities.

CONCLUSIONS: A mutual exclusion or a competition between Asaia and Wolbachia may contribute to explain the inability of Wolbachia to colonize the female reproductive organs of anophelines, inhibiting its vertical transmission and explaining the absence of Wolbachia infection in Ae. aegypti and in the majority of natural populations of Anopheles mosquitoes.}, } @article {pmid25972858, year = {2015}, author = {Strauß, JF and Telschow, A}, title = {Modeling the indirect effect of Wolbachia on the infection dynamics of horizontally transmitted viruses.}, journal = {Frontiers in microbiology}, volume = {6}, number = {}, pages = {378}, pmid = {25972858}, issn = {1664-302X}, abstract = {Intracellular bacteria of the genus Wolbachia are widely distributed in arthropods. There is growing empirical evidence that Wolbachia directly interacts with viruses and other parasites inside the arthropod host, sometimes resulting in low or no pathogen replication. Previous theoretical studies showed that this direct effect of Wolbachia can result in a reduced virus prevalence (within the population), suggesting that Wolbachia could be used in the biological control of vector-borne diseases (e.g., dengue fever). However, Wolbachia might also indirectly affect virus dynamics because Wolbachia-induced reproductive phenotypes (cytoplasmic incompatibility or male killing) increase the larval mortality of hosts and thus alter the age structure of populations. We investigated this indirect effect using mathematical models with overlapping generations, and found the results to depend strongly on the host's life history. In general, the indirect effect can result in two different outcomes: (1) reduced virus prevalence and virus invasion ability, and (2) increased virus prevalence and virus invasion ability. The former occurs for host species with larval competition and undercompensation, the latter for hosts with either adult competition or larval competition and overcompensation. These findings suggest that the effect of Wolbachia on a specific virus is sensitive to the host's life history. We discuss the results with respect to biocontrol programs using Wolbachia.}, } @article {pmid25963738, year = {2015}, author = {Ma, WJ and Pannebakker, BA and van de Zande, L and Schwander, T and Wertheim, B and Beukeboom, LW}, title = {Diploid males support a two-step mechanism of endosymbiont-induced thelytoky in a parasitoid wasp.}, journal = {BMC evolutionary biology}, volume = {15}, number = {}, pages = {84}, pmid = {25963738}, issn = {1471-2148}, mesh = {Animals ; Biological Evolution ; Diploidy ; Female ; Haploidy ; Male ; Mutation ; *Parthenogenesis ; *Sex Determination Processes ; Wasps/*genetics/*microbiology/physiology ; Wolbachia/genetics ; }, abstract = {BACKGROUND: Haplodiploidy, where females develop from diploid, fertilized eggs and males from haploid, unfertilized eggs, is abundant in some insect lineages. Some species in these lineages reproduce by thelytoky that is caused by infection with endosymbionts: infected females lay haploid eggs that undergo diploidization and develop into females, while males are very rare or absent. It is generally assumed that in thelytokous wasps, endosymbionts merely diploidize the unfertilized eggs, which would then trigger female development.

RESULTS: We found that females in the parasitoid wasp Asobara japonica infected with thelytoky-inducing Wolbachia produce 0.7-1.2% male offspring. Seven to 39% of these males are diploid, indicating that diploidization and female development can be uncoupled in A. japonica. Wolbachia titer in adults was correlated with their ploidy and sex: diploids carried much higher Wolbachia titers than haploids, and diploid females carried more Wolbachia than diploid males. Data from introgression lines indicated that the development of diploid individuals into males instead of females is not caused by malfunction-mutations in the host genome but that diploid males are most likely produced when the endosymbiont fails to activate the female sex determination pathway. Our data therefore support a two-step mechanism by which endosymbionts induce thelytoky in A. japonica: diploidization of the unfertilized egg is followed by feminization, whereby each step correlates with a threshold of endosymbiont titer during wasp development.

CONCLUSIONS: Our new model of endosymbiont-induced thelytoky overthrows the view that certain sex determination mechanisms constrain the evolution of endosymbiont-induced thelytoky in hymenopteran insects. Endosymbionts can cause parthenogenesis through feminization, even in groups in which endosymbiont-diploidized eggs would develop into males following the hosts' sex determination mechanism. In addition, our model broadens our understanding of the mechanisms by which endosymbionts induce thelytoky to enhance their transmission to the next generation. Importantly, it also provides a novel window to study the yet-poorly known haplodiploid sex determination mechanisms in haplodiploid insects.}, } @article {pmid25961733, year = {2015}, author = {Serbus, LR and White, PM and Silva, JP and Rabe, A and Teixeira, L and Albertson, R and Sullivan, W}, title = {Correction: the impact of host diet on wolbachia titer in Drosophila.}, journal = {PLoS pathogens}, volume = {11}, number = {5}, pages = {e1004889}, pmid = {25961733}, issn = {1553-7374}, } @article {pmid25948567, year = {2015}, author = {Kern, P and Cook, JM and Kageyama, D and Riegler, M}, title = {Double trouble: combined action of meiotic drive and Wolbachia feminization in Eurema butterflies.}, journal = {Biology letters}, volume = {11}, number = {5}, pages = {20150095}, pmid = {25948567}, issn = {1744-957X}, mesh = {Animals ; Butterflies/embryology/*genetics/growth & development/*microbiology ; Chromosome Segregation ; Female ; Male ; *Meiosis ; Real-Time Polymerase Chain Reaction ; Sex Chromosomes ; Sex Determination Processes/*physiology ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Arthropod sex ratios can be manipulated by a diverse range of selfish genetic elements, including maternally inherited Wolbachia bacteria. Feminization by Wolbachia is rare but has been described for Eurema mandarina butterflies. In this species, some phenotypic and functional females, thought to be ZZ genetic males, are infected with a feminizing Wolbachia strain, wFem. Meanwhile, heterogametic WZ females are not infected with wFem. Here, we establish a quantitative PCR assay allowing reliable sexing in three Eurema species. Against expectation, all E. mandarina females, including wFem females, had only one Z chromosome that was paternally inherited. Observation of somatic interphase nuclei confirmed that W chromatin was absent in wFem females, but present in females without wFem. We conclude that the sex bias in wFem lines is due to meiotic drive (MD) that excludes the maternal Z and thus prevents formation of ZZ males. Furthermore, wFem lines may have lost the W chromosome or harbour a dysfunctional version, yet rely on wFem for female development; removal of wFem results in all-male offspring. This is the first study that demonstrates an interaction between MD and Wolbachia feminization, and it highlights endosymbionts as potentially confounding factors in MD of sex chromosomes.}, } @article {pmid25948064, year = {2015}, author = {Debrah, AY and Specht, S and Klarmann-Schulz, U and Batsa, L and Mand, S and Marfo-Debrekyei, Y and Fimmers, R and Dubben, B and Kwarteng, A and Osei-Atweneboana, M and Boakye, D and Ricchiuto, A and Büttner, M and Adjei, O and Mackenzie, CD and Hoerauf, A}, title = {Doxycycline Leads to Sterility and Enhanced Killing of Female Onchocerca volvulus Worms in an Area With Persistent Microfilaridermia After Repeated Ivermectin Treatment: A Randomized, Placebo-Controlled, Double-Blind Trial.}, journal = {Clinical infectious diseases : an official publication of the Infectious Diseases Society of America}, volume = {61}, number = {4}, pages = {517-526}, pmid = {25948064}, issn = {1537-6591}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/*administration & dosage ; Double-Blind Method ; Doxycycline/*administration & dosage ; Female ; Filaricides/administration & dosage ; Ghana ; Humans ; Ivermectin/administration & dosage ; Male ; Middle Aged ; Onchocerca volvulus/*drug effects/microbiology/*physiology ; Onchocerciasis/*drug therapy ; Placebos/administration & dosage ; Treatment Outcome ; Wolbachia/*drug effects ; Young Adult ; }, abstract = {BACKGROUND: Ivermectin (IVM) has been the drug of choice for the treatment of onchocerciasis. However, there have been reports of persistent microfilaridermia in individuals from an endemic area in Ghana after many rounds of IVM, raising concerns of suboptimal response or even the emergence of drug resistance. Because it is considered risky to continue relying only on IVM to combat this phenomenon, we assessed the effect of targeting the Onchocerca volvulus Wolbachia endosymbionts with doxycycline for these individuals with suboptimal response.

METHODS: One hundred sixty-seven patients, most of them with multiple rounds of IVM, were recruited in areas with IVM suboptimal response and treated with 100 mg/day doxycycline for 6 weeks. Three and 12 months after doxycycline treatment, patients took part in standard IVM treatment.

RESULTS: At 20 months after treatment, 80% of living female worms from the placebo group were Wolbachia positive, whereas only 5.1% in the doxycycline-treated group contained bacteria. Consistent with interruption of embryogenesis, none of the nodules removed from doxycycline-treated patients contained microfilariae, and 97% of those patients were without microfilaridermia, in contrast to placebo patients who remained at pretreatment levels (P < .001). Moreover, a significantly enhanced number of dead worms were observed after doxycycline.

CONCLUSIONS: Targeting the Wolbachia in O. volvulus is effective in clearing microfilariae in the skin of onchocerciasis patients with persistent microfilaridermia and in enhanced killing of adult worms after repeated standard IVM treatment. Strategies can now be developed that include doxycycline to control onchocerciasis in areas where infections persist despite the frequent use of IVM.

CLINICAL TRIALS REGISTRATION: ISRCTN 66649839.}, } @article {pmid25934616, year = {2015}, author = {Rohrscheib, CE and Bondy, E and Josh, P and Riegler, M and Eyles, D and van Swinderen, B and Weible, MW and Brownlie, JC}, title = {Wolbachia Influences the Production of Octopamine and Affects Drosophila Male Aggression.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {14}, pages = {4573-4580}, pmid = {25934616}, issn = {1098-5336}, mesh = {Animals ; Behavior, Animal ; Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster/enzymology/genetics/*microbiology/*physiology ; Female ; Male ; Mixed Function Oxygenases/genetics/metabolism ; Octopamine/*biosynthesis ; Tyrosine Decarboxylase/genetics/metabolism ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are endosymbionts that infect approximately 40% of all insect species and are best known for their ability to manipulate host reproductive systems. Though the effect Wolbachia infection has on somatic tissues is less well understood, when present in cells of the adult Drosophila melanogaster brain, Wolbachia exerts an influence over behaviors related to olfaction. Here, we show that a strain of Wolbachia influences male aggression in flies, which is critically important in mate competition. A specific strain of Wolbachia was observed to reduce the initiation of aggressive encounters in Drosophila males compared to the behavior of their uninfected controls. To determine how Wolbachia was able to alter aggressive behavior, we investigated the role of octopamine, a neurotransmitter known to influence male aggressive behavior in many insect species. Transcriptional analysis of the octopamine biosynthesis pathway revealed that two essential genes, the tyrosine decarboxylase and tyramine β-hydroxylase genes, were significantly downregulated in Wolbachia-infected flies. Quantitative chemical analysis also showed that total octopamine levels were significantly reduced in the adult heads.}, } @article {pmid25933928, year = {2015}, author = {Ghosh, S and Bouvaine, S and Maruthi, MN}, title = {Prevalence and genetic diversity of endosymbiotic bacteria infecting cassava whiteflies in Africa.}, journal = {BMC microbiology}, volume = {15}, number = {}, pages = {93}, pmid = {25933928}, issn = {1471-2180}, mesh = {Africa ; Animals ; Bacteria/*classification/genetics/*isolation & purification ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Mitochondrial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; *Genetic Variation ; Hemiptera/*microbiology ; Manihot/parasitology ; Molecular Sequence Data ; Phylogeny ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 23S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {BACKGROUND: Cassava provides over half of the dietary requirement for more than 200 million poor in Africa. In recent years, cassava has been affected by an epidemic of a virus disease called cassava brown streak disease (CBSD) that is spreading in much of eastern and central Africa, affecting food security and the economic development of the poor. The viruses that cause CBSD are transmitted by the insect vector whitefly (Bemisia tabaci), which have increased to very high numbers in some African countries. Strains of endosymbiotic bacteria infecting whiteflies have been reported to interact specifically with different whitefly populations with varied effects on its host biology and efficiency of virus transmission. The main aim of this study was therefore to investigate the prevalence and diversity of the secondary endosymbiotic bacteria infecting cassava whiteflies with a view to better understand their role on insect population dynamics and virus disease epidemics.

RESULTS: The genetic diversity of field-collected whitefly from Tanzania, Malawi, Uganda and Nigeria was determined by mitochondrial DNA based phylogeny and restriction fragment length polymorphism. Cassava in these countries was infected with five whitefly populations, and each one was infected with different endosymbiotic bacteria. Incidences of Arsenophonus, Rickettsia, Wolbachia and Cardinium varied amongst the populations. Wolbachia was the most predominant symbiont with infection levels varying from 21 to 97%. Infection levels of Arsenophonus varied from 17 to 64% and that of Rickettsia was 0 to 53%. Hamiltonella and Fritschea were absent in all the samples. Multiple locus sequence typing identified four different strains of Wolbachia infecting cassava whiteflies. A common strain of Wolbachia infected the whitefly population Sub-Saharan Africa 1-subgroup 1 (SSA1-SG1) and SSA1-SG2, while others were infected with different strains. Phylogeny based on 16S rDNA of Rickettsia and 23S rDNA of Arsenophonus also identified distinct strains.

CONCLUSIONS: Genetically diverse bacteria infect cassava whiteflies in Africa with varied prevalence across different host populations, which may affect their whitefly biology. Further studies are required to investigate the role of endosymbionts to better understand the whitefly population dynamics.}, } @article {pmid25923352, year = {2015}, author = {Raquin, V and Valiente Moro, C and Saucereau, Y and Tran, FH and Potier, P and Mavingui, P}, title = {Native Wolbachia from Aedes albopictus Blocks Chikungunya Virus Infection In Cellulo.}, journal = {PloS one}, volume = {10}, number = {4}, pages = {e0125066}, pmid = {25923352}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Animals ; Chikungunya Fever/microbiology/*transmission/virology ; Chikungunya virus/genetics/*pathogenicity ; Humans ; Symbiosis ; Virus Replication/genetics ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Wolbachia, a widespread endosymbiont of terrestrial arthropods, can protect its host against viral and parasitic infections, a phenotype called "pathogen blocking". However, in some cases Wolbachia may have no effect or even enhance pathogen infection, depending on the host-Wolbachia-pathogen combination. The tiger mosquito Aedes albopictus is naturally infected by two strains of Wolbachia, wAlbA and wAlbB, and is a competent vector for different arboviruses such as dengue virus (DENV) and chikungunya virus (CHIKV). Interestingly, it was shown in some cases that Ae. albopictus native Wolbachia strains are able to inhibit DENV transmission by limiting viral replication in salivary glands, but no such impact was measured on CHIKV replication in vivo. To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain. Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB. Using FISH, we observed CHIKV and wAlbB in the same mosquito cells, indicating that the virus is still able to enter the cell in the presence of the bacterium. Further work is needed to decipher molecular pathways involved in Wolbachia-CHIKV interaction at the cellular level, but this cellular model can be a useful tool to study the mechanism behind virus blocking phenotype induced by Wolbachia. More broadly, this put into question the ecological role of Wolbachia symbiont in Ae. albopictus, but also the ability of the CHIKV to counteract Wolbachia's antiviral potential in vivo.}, } @article {pmid25921489, year = {2015}, author = {Brown, AN and Lloyd, VK}, title = {Evidence for horizontal transfer of Wolbachia by a Drosophila mite.}, journal = {Experimental & applied acarology}, volume = {66}, number = {3}, pages = {301-311}, pmid = {25921489}, issn = {1572-9702}, mesh = {Acaridae/*microbiology/*physiology ; Animals ; Base Sequence ; DNA, Bacterial/*genetics ; Drosophila melanogaster/growth & development/*microbiology/*parasitology/physiology ; Feeding Behavior ; Female ; Larva/growth & development/microbiology/parasitology/physiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Alignment ; Wolbachia/genetics/*physiology ; }, abstract = {Mites are common ectoparasites of Drosophila and have been implicated in bacterial and mobile element invasion of Drosophila stocks. The obligate endobacterium, Wolbachia, has widespread effects on gene expression in their arthropod hosts and alters host reproduction to enhance its survival and propagation, often with deleterious effects in Drosophila hosts. To determine whether Wolbachia could be transferred between Drosophila melanogaster laboratory stocks by the mite Tyrophagus putrescentiae, mites were introduced to Wolbachia-infected Drosophila vials. These vials were kept adjacent to mite-free and Wolbachia-uninfected Drosophila stock vials. The Wolbachia infection statuses of the infected and uninfected flies were checked from generation 1 to 5. Results indicate that Wolbachia DNA could be amplified from mites infesting Wolbachia-infected fly stocks and infection in the previously uninfected stocks arose within generation 1 or 2, concomitant with invasion of mites from the Wolbachia-infected stock. A possible mechanism for the transfer of Wolbachia from flies to mites and vice versa, can be inferred from time-lapse photography of fly and mite interactions. We demonstrated that mites ingest Drosophila corpses, including Wolbachia-infected corpses, and Drosophila larva ingest mites, providing possible sources of Wolbachia infection and transfer. This research demonstrated that T. putrescentiae white mites can facilitate Wolbachia transfer between Drosophila stocks and that this may occur by ingestion of infected corpses. Mite-vectored Wolbachia transfer allows for rapid establishment of Wolbachia infection within a new population. This mode of Wolbachia introduction may be relevant in nature as well as in the laboratory, and could have a variety of biological consequences.}, } @article {pmid25914139, year = {2015}, author = {Gerth, M and Saeed, A and White, JA and Bleidorn, C}, title = {Extensive screen for bacterial endosymbionts reveals taxon-specific distribution patterns among bees (Hymenoptera, Anthophila).}, journal = {FEMS microbiology ecology}, volume = {91}, number = {6}, pages = {}, doi = {10.1093/femsec/fiv047}, pmid = {25914139}, issn = {1574-6941}, mesh = {Animals ; Bacteroidetes/genetics/isolation & purification ; Bees/*microbiology ; Enterobacteriaceae/genetics/isolation & purification ; Female ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny ; Rickettsia/genetics/isolation & purification ; Symbiosis/*physiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Bacterial endosymbionts play key roles in arthropod biology, ranging from beneficial mutualists to parasitic sex ratio manipulators. The number of described endosymbiotic bacterial taxa has accumulated continuously in recent years. While the understanding of arthropod-microbe interactions has advanced significantly, especially in model organisms, relatively little is known about symbiont distribution and effects in non-model organisms. As a first step to alleviate this gap in understanding, we performed an endosymbiont survey in bees (Anthophila), an ecologically and economically important group of hymenopterans. To this end, we sampled 170 bee species and screened by PCR for the presence of Wolbachia, Rickettsia, Arsenophonus and Cardinium. Detected strains were then further diagnosed by additional markers. Additionally, we tested if certain ecological traits, bee phylogeny or geographic origin of bees explain endosymbiont distribution. Our results indicate that supergroup A Wolbachia are very common in bees and that their distribution can be significantly correlated to both host ecology and phylogeny, although a distinction of these factors is not possible. Furthermore, bees from the same region (Old World or New World) are more likely to harbour identical Wolbachia strains than expected by chance. Other endosymbionts (Rickettsia, Arsenophonus) were less common, and specific to particular host taxa, suggesting that host phylogeny is a major predictor for endosymbiont distribution in bees.}, } @article {pmid25907143, year = {2015}, author = {Liberti, J and Sapountzis, P and Hansen, LH and Sørensen, SJ and Adams, RM and Boomsma, JJ}, title = {Bacterial symbiont sharing in Megalomyrmex social parasites and their fungus-growing ant hosts.}, journal = {Molecular ecology}, volume = {24}, number = {12}, pages = {3151-3169}, pmid = {25907143}, issn = {1365-294X}, support = {323085/ERC_/European Research Council/International ; }, mesh = {Actinomycetales/*classification ; Animals ; Ants/classification/*microbiology ; DNA, Bacterial/genetics ; Fungi ; Genotype ; *Microbiota ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; }, abstract = {Bacterial symbionts are important fitness determinants of insects. Some hosts have independently acquired taxonomically related microbes to meet similar challenges, but whether distantly related hosts that live in tight symbiosis can maintain similar microbial communities has not been investigated. Varying degrees of nest sharing between Megalomyrmex social parasites (Solenopsidini) and their fungus-growing ant hosts (Attini) from the genera Cyphomyrmex, Trachymyrmex and Sericomyrmex allowed us to address this question, as both ant lineages rely on the same fungal diet, interact in varying intensities and are distantly related. We used tag-encoded FLX 454 pyrosequencing and diagnostic PCR to map bacterial symbiont diversity across the Megalomyrmex phylogenetic tree, which also contains free-living generalist predators. We show that social parasites and hosts share a subset of bacterial symbionts, primarily consisting of Entomoplasmatales, Bartonellaceae, Acinetobacter, Wolbachia and Pseudonocardia and that Entomoplasmatales and Bartonellaceae can co-infect specifically associated combinations of hosts and social parasites with identical 16S rRNA genotypes. We reconstructed in more detail the population-level infection dynamics for Entomoplasmatales and Bartonellaceae in Megalomyrmex symmetochus guest ants and their Sericomyrmex amabilis hosts. We further assessed the stability of the bacterial communities through a diet manipulation experiment and evaluated possible transmission modes in shared nests such as consumption of the same fungus garden food, eating of host brood by social parasites, trophallaxis and grooming interactions between the ants, or parallel acquisition from the same nest environment. Our results imply that cohabiting ant social parasites and hosts may obtain functional benefits from bacterial symbiont transfer even when they are not closely related.}, } @article {pmid25906323, year = {2015}, author = {Villela, DA and Codeço, CT and Figueiredo, F and Garcia, GA and Maciel-de-Freitas, R and Struchiner, CJ}, title = {A Bayesian Hierarchical Model for Estimation of Abundance and Spatial Density of Aedes aegypti.}, journal = {PloS one}, volume = {10}, number = {4}, pages = {e0123794}, pmid = {25906323}, issn = {1932-6203}, mesh = {*Aedes ; Animals ; *Bayes Theorem ; Brazil ; *Models, Theoretical ; Population Density ; }, abstract = {Strategies to minimize dengue transmission commonly rely on vector control, which aims to maintain Ae. aegypti density below a theoretical threshold. Mosquito abundance is traditionally estimated from mark-release-recapture (MRR) experiments, which lack proper analysis regarding accurate vector spatial distribution and population density. Recently proposed strategies to control vector-borne diseases involve replacing the susceptible wild population by genetically modified individuals' refractory to the infection by the pathogen. Accurate measurements of mosquito abundance in time and space are required to optimize the success of such interventions. In this paper, we present a hierarchical probabilistic model for the estimation of population abundance and spatial distribution from typical mosquito MRR experiments, with direct application to the planning of these new control strategies. We perform a Bayesian analysis using the model and data from two MRR experiments performed in a neighborhood of Rio de Janeiro, Brazil, during both low- and high-dengue transmission seasons. The hierarchical model indicates that mosquito spatial distribution is clustered during the winter (0.99 mosquitoes/premise 95% CI: 0.80-1.23) and more homogeneous during the high abundance period (5.2 mosquitoes/premise 95% CI: 4.3-5.9). The hierarchical model also performed better than the commonly used Fisher-Ford's method, when using simulated data. The proposed model provides a formal treatment of the sources of uncertainty associated with the estimation of mosquito abundance imposed by the sampling design. Our approach is useful in strategies such as population suppression or the displacement of wild vector populations by refractory Wolbachia-infected mosquitoes, since the invasion dynamics have been shown to follow threshold conditions dictated by mosquito abundance. The presence of spatially distributed abundance hotspots is also formally addressed under this modeling framework and its knowledge deemed crucial to predict the fate of transmission control strategies based on the replacement of vector populations.}, } @article {pmid25906062, year = {2015}, author = {Newton, IL and Savytskyy, O and Sheehan, KB}, title = {Wolbachia utilize host actin for efficient maternal transmission in Drosophila melanogaster.}, journal = {PLoS pathogens}, volume = {11}, number = {4}, pages = {e1004798}, pmid = {25906062}, issn = {1553-7374}, support = {P40 OD018537/OD/NIH HHS/United States ; NIH P40OD018537/OD/NIH HHS/United States ; }, mesh = {Actins/metabolism ; Animals ; Blotting, Western ; Drosophila melanogaster/*parasitology ; Host-Parasite Interactions/*physiology ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Infectious Disease Transmission, Vertical ; Phenotype ; Polymerase Chain Reaction ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia pipientis is a ubiquitous, maternally transmitted bacterium that infects the germline of insect hosts. Estimates are that Wolbachia infect nearly 40% of insect species on the planet, making it the most prevalent infection on Earth. The bacterium, infamous for the reproductive phenotypes it induces in arthropod hosts, has risen to recent prominence due to its use in vector control. Wolbachia infection prevents the colonization of vectors by RNA viruses, including Drosophila C virus and important human pathogens such as Dengue and Chikungunya. Here we present data indicating that Wolbachia utilize the host actin cytoskeleton during oogenesis for persistence within and transmission between Drosophila melanogaster generations. We show that phenotypically wild type flies heterozygous for cytoskeletal mutations in Drosophila profilin (chic(221/+) and chic(1320/+)) or villin (qua(6-396/+)) either clear a Wolbachia infection, or result in significantly reduced infection levels. This reduction of Wolbachia is supported by PCR evidence, Western blot results and cytological examination. This phenotype is unlikely to be the result of maternal loading defects, defects in oocyte polarization, or germline stem cell proliferation, as the flies are phenotypically wild type in egg size, shape, and number. Importantly, however, heterozygous mutant flies exhibit decreased total G-actin in the ovary, compared to control flies and chic(221) heterozygous mutants exhibit decreased expression of profilin. Additionally, RNAi knockdown of profilin during development decreases Wolbachia titers. We analyze evidence in support of alternative theories to explain this Wolbachia phenotype and conclude that our results support the hypothesis that Wolbachia utilize the actin skeleton for efficient transmission and maintenance within Drosophila.}, } @article {pmid25905888, year = {2015}, author = {Dutra, HL and Dos Santos, LM and Caragata, EP and Silva, JB and Villela, DA and Maciel-de-Freitas, R and Moreira, LA}, title = {From lab to field: the influence of urban landscapes on the invasive potential of Wolbachia in Brazilian Aedes aegypti mosquitoes.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {4}, pages = {e0003689}, pmid = {25905888}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Brazil ; *Cities ; Dengue/*prevention & control/transmission ; *Environment ; Fertility/physiology ; Humans ; Models, Theoretical ; Pest Control, Biological/*methods ; *Phenotype ; Wolbachia/*genetics/physiology ; }, abstract = {BACKGROUND: The symbiotic bacterium Wolbachia is currently being trialled as a biocontrol agent in several countries to reduce dengue transmission. Wolbachia can invade and spread to infect all individuals within wild mosquito populations, but requires a high rate of maternal transmission, strong cytoplasmic incompatibility and low fitness costs in the host in order to do so. Additionally, extensive differences in climate, field-release protocols, urbanization level and human density amongst the sites where this bacterium has been deployed have limited comparison and analysis of Wolbachia's invasive potential.

We examined key phenotypic effects of the wMel Wolbachia strain in laboratory Aedes aegypti mosquitoes with a Brazilian genetic background to characterize its invasive potential. We show that the wMel strain causes strong cytoplasmic incompatibility, a high rate of maternal transmission and has no evident detrimental effect on host fecundity or fertility. Next, to understand the effects of different urban landscapes on the likelihood of mosquito survival, we performed mark-release-recapture experiments using Wolbachia-uninfected Brazilian mosquitoes in two areas of Rio de Janeiro where Wolbachia will be deployed in the future. We characterized the mosquito populations in relation to the socio-demographic conditions at these sites, and at three other future release areas. We then constructed mathematical models using both the laboratory and field data, and used these to describe the influence of urban environmental conditions on the likelihood that the Wolbachia infection frequency could reach 100% following mosquito release. We predict successful invasion at all five field sites, however the conditions by which this occurs vary greatly between sites, and are strongly influenced by the size of the local mosquito population.

CONCLUSIONS/SIGNIFICANCE: Through analysis of laboratory, field and mathematical data, we show that the wMel strain of Wolbachia possesses the characteristics required to spread effectively in different urban socio-demographic environments in Rio de Janeiro, including those where mosquito releases from the Eliminate Dengue Program will take place.}, } @article {pmid25904667, year = {2015}, author = {Weinert, LA and Araujo-Jnr, EV and Ahmed, MZ and Welch, JJ}, title = {The incidence of bacterial endosymbionts in terrestrial arthropods.}, journal = {Proceedings. Biological sciences}, volume = {282}, number = {1807}, pages = {20150249}, pmid = {25904667}, issn = {1471-2954}, mesh = {Animals ; Arthropods/*microbiology ; Bacteroidetes/*physiology ; Incidence ; Likelihood Functions ; Rickettsia/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Intracellular endosymbiotic bacteria are found in many terrestrial arthropods and have a profound influence on host biology. A basic question about these symbionts is why they infect the hosts that they do, but estimating symbiont incidence (the proportion of potential host species that are actually infected) is complicated by dynamic or low prevalence infections. We develop a maximum-likelihood approach to estimating incidence, and testing hypotheses about its variation. We apply our method to a database of screens for bacterial symbionts, containing more than 3600 distinct arthropod species and more than 150 000 individual arthropods. After accounting for sampling bias, we estimate that 52% (CIs: 48-57) of arthropod species are infected with Wolbachia, 24% (CIs: 20-42) with Rickettsia and 13% (CIs: 13-55) with Cardinium. We then show that these differences stem from the significantly reduced incidence of Rickettsia and Cardinium in most hexapod orders, which might be explained by evolutionary differences in the arthropod immune response. Finally, we test the prediction that symbiont incidence should be higher in speciose host clades. But while some groups do show a trend for more infection in species-rich families, the correlations are generally weak and inconsistent. These results argue against a major role for parasitic symbionts in driving arthropod diversification.}, } @article {pmid25903749, year = {2015}, author = {Amuzu, HE and Simmons, CP and McGraw, EA}, title = {Effect of repeat human blood feeding on Wolbachia density and dengue virus infection in Aedes aegypti.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {246}, pmid = {25903749}, issn = {1756-3305}, support = {089276//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; *Bacterial Load ; Blood/*metabolism ; *Feeding Behavior ; Gastrointestinal Tract/microbiology/virology ; Humans ; Salivary Glands/microbiology/virology ; *Viral Load ; Wolbachia/growth & development/*isolation & purification ; }, abstract = {BACKGROUND: The introduction of the endosymbiotic bacterium, Wolbachia into Aedes aegypti populations is a novel approach to reduce disease transmission. The presence of Wolbachia limits the ability of the mosquito to transmit dengue virus (DENV) and the strength of this effect appears to correlate with Wolbachia densities in the mosquito. There is also some evidence that Wolbachia densities may increase following the consumption of a bloodmeal. Here we have examined whether multiple blood feeds lead to increases in density or associated changes in Wolbachia-mediated blocking of DENV.

METHODS: The Wolbachia infected Aedes aegypti mosquito line was used for the study. There were three treatment groups; a non-blood fed control, a second group fed once and a third group fed twice on human blood. All groups were orally infected with DENV-2 and then their midguts and salivary glands were dissected 10-11 days post infection. RNA/DNA was simultaneously extracted from each tissue and subsequently used for DENV RNA copies and Wolbachia density quantification, respectively.

RESULTS: We found variation between replicate vector competence experiments and no clear evidence that Wolbachia numbers increased in either the salivary glands or remainder of the body with feeding and hence saw no corresponding improvements in DENV blocking.

CONCLUSIONS: Aedes aegypti are "sip" feeders returning often to obtain bloodmeals and hence it is important to assess whether repeat blood feeding improved the efficacy of Wolbachia-based DENV blocking. Our work suggests in the laboratory context when Wolbachia densities are high that repeat feeding does not improve blocking and hence this ability should likely be stable with respect to feeding cycle in the field.}, } @article {pmid25903547, year = {2015}, author = {Yan, Q and Qiao, H and Gao, J and Yun, Y and Liu, F and Peng, Y}, title = {Detection and phylogenetic analysis of bacteriophage WO in spiders (Araneae).}, journal = {Folia microbiologica}, volume = {60}, number = {6}, pages = {497-503}, pmid = {25903547}, issn = {1874-9356}, mesh = {Animals ; Bacteriophages/*classification/genetics/*isolation & purification ; Molecular Sequence Data ; *Phylogeny ; Spiders/classification/*virology ; Wolbachia/virology ; }, abstract = {Phage WO is a bacteriophage found in Wolbachia. Herein, we represent the first phylogenetic study of WOs that infect spiders (Araneae). Seven species of spiders (Araneus alternidens, Nephila clavata, Hylyphantes graminicola, Prosoponoides sinensis, Pholcus crypticolens, Coleosoma octomaculatum, and Nurscia albofasciata) from six families were infected by Wolbachia and WO, followed by comprehensive sequence analysis. Interestingly, WO could be only detected Wolbachia-infected spiders. The relative infection rates of those seven species of spiders were 75, 100, 88.9, 100, 62.5, 72.7, and 100 %, respectively. Our results indicated that both Wolbachia and WO were found in three different body parts of N. clavata, and WO could be passed to the next generation of H. graminicola by vertical transmission. There were three different sequences for WO infected in A. alternidens and two different WO sequences from C. octomaculatum. Only one sequence of WO was found for the other five species of spiders. The discovered sequence of WO ranged from 239 to 311 bp. Phylogenetic tree was generated using maximum likelihood (ML) based on the orf7 gene sequences. According to the phylogenetic tree, WOs in N. clavata and H. graminicola were clustered in the same group. WOs from A. alternidens (WAlt1) and C. octomaculatum (WOct2) were closely related to another clade, whereas WO in P. sinensis was classified as a sole cluster.}, } @article {pmid25899523, year = {2015}, author = {Joanne, S and Vythilingam, I and Yugavathy, N and Leong, CS and Wong, ML and AbuBakar, S}, title = {Distribution and dynamics of Wolbachia infection in Malaysian Aedes albopictus.}, journal = {Acta tropica}, volume = {148}, number = {}, pages = {38-45}, doi = {10.1016/j.actatropica.2015.04.003}, pmid = {25899523}, issn = {1873-6254}, mesh = {Aedes/*microbiology ; Animals ; Dengue/*epidemiology/transmission ; Genetic Variation ; Genotype ; Humans ; Insect Vectors/*microbiology ; Malaysia/epidemiology ; Polymerase Chain Reaction ; Wolbachia/*genetics/physiology ; }, abstract = {Wolbachia are maternally transmitted bacteria found in most arthropods and nematodes, but little is known about their distribution and reproductive dynamics in the Malaysian dengue vector Aedes albopictus. In this study, polymerase chain reaction (PCR) was used to determine the presence of Wolbachia from field collected Ae. albopictus from various parts of the country using wsp specific primers. Ae. albopictus had Wolbachia infection ranging from 60 to 100%. No sequence diversity of wsp gene was found within all wAlbA and wAlbB sequences. Our findings suggest that Wolbachia infection amongst the Malaysian Ae. albopictus were not homogenously distributed in all districts in Malaysia. The presence of Wolbachia in different organs of Ae. albopictus was also determined. Wolbachia were only found in the ovaries and midguts of the mosquitoes, while absent in the salivary glands. The effects of Wolbachia on Ae. albopictus fecundity, longevity and egg viability were studied using infected and uninfected colonies. The removal of Wolbachia from Ae. albopictus resulted in reduced fecundity, longevity and egg viability, thus. Wolbachia seem to play a vital role in Ae. albopictus reproductive system.}, } @article {pmid25899011, year = {2015}, author = {Curry, MM and Paliulis, LV and Welch, KD and Harwood, JD and White, JA}, title = {Multiple endosymbiont infections and reproductive manipulations in a linyphiid spider population.}, journal = {Heredity}, volume = {115}, number = {2}, pages = {146-152}, pmid = {25899011}, issn = {1365-2540}, mesh = {Animals ; Female ; Fertility ; Male ; Phenotype ; *Rickettsia ; *Sex Ratio ; Spiders/*genetics/*microbiology ; *Symbiosis ; *Wolbachia ; }, abstract = {In many arthropods, maternally inherited endosymbiotic bacteria can increase infection frequency by manipulating host reproduction. Multiple infections of different bacteria in a single host population are common, yet few studies have documented concurrent endosymbiont phenotypes or explored their potential interactions. We hypothesized that spiders might be a particularly useful taxon for investigating endosymbiont interactions, because they are host to a plethora of endosymbiotic bacteria and frequently exhibit multiple infections. We established two matrilines from the same population of the linyphiid spider Mermessus fradeorum and then used antibiotic curing and controlled mating assays to demonstrate that each matriline was subject to a distinct endosymbiotic reproductive manipulation. One matriline was co-infected with Rickettsia and Wolbachia and produced offspring with a radical female bias. Antibiotic treatment eliminated both endosymbionts and restored an even sex ratio to subsequent generations. Chromosomal and fecundity observations suggest a feminization mechanism. In the other matriline, a separate factorial mating assay of cured and infected spiders demonstrated strong cytoplasmic incompatibility (CI) induced by a different strain of Wolbachia. However, males with this Wolbachia induced only mild CI when mated with the Rickettsia-Wolbachia females. In a subsequent survey of a field population of M. fradeorum, we detected these same three endosymbionts infecting 55% of the spiders in almost all possible combinations, with nearly half of the infected spiders exhibiting multiple infection. Our results suggest that a dynamic network of endosymbionts may interact both within multiply infected hosts and within a population subject to multiple strong reproductive manipulations.}, } @article {pmid25886525, year = {2015}, author = {Maia, C and Almeida, B and Coimbra, M and Fernandes, MC and Cristóvão, JM and Ramos, C and Martins, Â and Martinho, F and Silva, P and Neves, N and Nunes, M and Vieira, ML and Cardoso, L and Campino, L}, title = {Bacterial and protozoal agents of canine vector-borne diseases in the blood of domestic and stray dogs from southern Portugal.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {138}, pmid = {25886525}, issn = {1756-3305}, mesh = {Animals ; Bacteria/classification/*isolation & purification ; Bacterial Infections/epidemiology/microbiology/*veterinary ; Blood/*microbiology/*parasitology ; Disease Transmission, Infectious ; Dog Diseases/*epidemiology/microbiology/parasitology ; Dogs ; Molecular Diagnostic Techniques ; Parasites/classification/*isolation & purification ; Polymerase Chain Reaction ; Portugal/epidemiology ; Protozoan Infections, Animal/epidemiology/*parasitology ; Risk Assessment ; Zoonoses/transmission ; }, abstract = {BACKGROUND: The so-called canine vector-borne diseases (CVBD) are caused by a wide range of pathogens transmitted by arthropods. In addition to their veterinary importance, many of these canine vector-borne pathogens can also affect the human population due to their zoonotic potential, a situation that requires a One Health approach. As the prevalence of vector-borne pathogens in cats from southern Portugal has been recently evaluated, the aim of the present study was to assess if the same agents were present in dogs living in the same area, and to assess positivity-associated risk factors.

METHODS: One thousand and ten dogs (521 domestic and 489 stray) from veterinary medical centres and animal shelters in southern Portugal were enrolled. Anaplasma spp./Ehrlichia spp., Bartonella spp., Borrelia burgdorferi sensu lato, Babesia spp., Hepatozoon spp. and Leishmania infantum infections were evaluated by polymerase chain reaction (PCR) assays in blood samples.

RESULTS: Sixty-eight (6.7%) dogs were PCR-positive to at least one of the tested CVBD agent species, genera or complex, including one dog found positive to two different genera. Nineteen (1.9%) dogs were positive to Anaplasma spp./Ehrlichia spp., eight (0.8%) to B. burgdorferi s.l., 31 (3.1%) to Hepatozoon spp. and 11 (1.1%) to L. infantum. Anaplasma platys, Ehrlichia canis, B. burgdorferis.l. and Hepatozoon canis were identified by DNA sequencing, including one animal confirmed with both A. platys and H. canis. Furthermore, Wolbachia spp. was amplified in blood from four dogs. None of the tested dogs was positive by PCR for Bartonella spp. or Babesia spp.

CONCLUSIONS: The molecular identification of CVBD agents in southern Portugal, some of them with zoonotic concern, reinforces the importance to alert the veterinary community, owners and public health authorities to prevent the risk of transmission of vector-borne pathogens among dogs and to other vertebrate hosts including humans. The prevalence of the selected pathogens was lower than that previously found in cats from the same region, probably because veterinarians and owners are more aware of them in the canine population and control measures are used more often.}, } @article {pmid25880387, year = {2015}, author = {Nguyen, DT and Spooner-Hart, RN and Riegler, M}, title = {Polyploidy versus endosymbionts in obligately thelytokous thrips.}, journal = {BMC evolutionary biology}, volume = {15}, number = {}, pages = {23}, pmid = {25880387}, issn = {1471-2148}, mesh = {Animals ; Cell Nucleus/genetics ; Female ; Genetic Variation ; Male ; Mitochondria/genetics ; *Parthenogenesis ; Phylogeny ; Polymerase Chain Reaction ; *Polyploidy ; Symbiosis ; Thysanoptera/classification/*genetics/*microbiology/physiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Thelytoky, the parthenogenetic development of females, has independently evolved in several insect orders yet the study of its mechanisms has so far mostly focussed on haplodiploid Hymenoptera, while alternative mechanisms of thelytoky such as polyploidy are far less understood. In haplodiploid insects, thelytoky can be encoded in their genomes, or induced by maternally inherited bacteria such as Wolbachia or Cardinium. Microbially facilitated thelytoky usually results in complete homozygosity due to gamete duplication and can be reverted into arrhenotoky, the parthenogenetic development of males, through treatment with antibiotics. In contrast, genetically encoded thelytoky cannot be removed and may result in conservation of heterozygosity due to gamete fusion. We have probed the obligate thelytoky of the greenhouse thrips, Heliothrips haemorrhoidalis (Bouché), a significant cosmopolitan pest and a model species of thelytoky in the haplodiploid insect order Thysanoptera. Earlier studies suggested terminal fusion as a mechanism for thelytoky in this species, while another study reported presence of Wolbachia; later it was speculated that Wolbachia plays a role in this thrips' thelytokous reproduction.

RESULTS: By using PCR and sequence analysis, we demonstrated that global population samples of H. haemorrhoidalis were not infected with Wolbachia, Cardinium or any other known bacterial reproductive manipulators. Antibiotic treatment of this thrips did also not result in male production. Some individuals carried two different alleles in two nuclear loci, histone 3 and elongation factor 1 alpha, suggesting heterozygosity. However, the majority of individuals had three different alleles suggesting that they were polyploid. Genetic diversity across both nuclear loci was low in all populations, and absent from mitochondrial cytochrome oxidase I, indicating that this species had experienced genetic bottlenecks, perhaps due to its invasion biology or a switch to thelytoky.

CONCLUSIONS: Geographically broad sampling and experimental manipulation revealed low genetic diversity, absence of Wolbachia but presence of three different alleles of nuclear loci in most analysed individuals of obligately thelytokous H. haemorrhoidalis. This suggests that polyploidy may be involved in the thelytokous reproduction of this thrips species, and polyploidy may be a contributing factor in the reproduction of Thysanoptera and other haplodiploid insect orders.}, } @article {pmid25875485, year = {2015}, author = {Kolopack, PA and Parsons, JA and Lavery, JV}, title = {What makes community engagement effective?: Lessons from the Eliminate Dengue Program in Queensland Australia.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {4}, pages = {e0003713}, pmid = {25875485}, issn = {1935-2735}, mesh = {Adult ; Australia ; *Community-Institutional Relations ; Dengue/epidemiology/*prevention & control ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Information Dissemination/methods ; Male ; Mosquito Control/methods ; Public Opinion ; Queensland/epidemiology ; Wolbachia ; }, abstract = {BACKGROUND: Worldwide, more than 40% of the population is at risk from dengue and recent estimates suggest that up to 390 million dengue infections are acquired every year. The Eliminate Dengue (ED) Program is investigating the use of Wolbachia-infected, transmission-compromised, mosquitoes to reduce dengue transmission. Previous introductions of genetically-modified strategies for dengue vector control have generated controversy internationally by inadequately engaging host communities. Community Engagement (CE) was a key component of the ED Program's initial open release trials in Queensland Australia. Their approach to CE was perceived as effective by the ED team's senior leadership, members of its CE team, and by its funders, but if and why this was the case was unclear. We conducted a qualitative case study of the ED Program's approach to CE to identify and critically examine its components, and to explain whether and how these efforts contributed to the support received by stakeholders.

In-depth semi-structured interviews were conducted with 24 participants with a range of experiences and perspectives related to the ED Program's CE activities. Our analytic approach combined techniques of grounded theory and qualitative description. The ED Program's approach to CE reflected four foundational features: 1) enabling conditions; 2) leadership; 3) core commitments and guiding values; and 4) formative social science research. These foundations informed five key operational practices: 1) building the CE team; 2) integrating CE into management practices; 3) discerning the community of stakeholders; 4) establishing and maintaining a presence in the community; and 5) socializing the technology and research strategy. We also demonstrate how these practices contributed to stakeholders' willingness to support the trials.

CONCLUSIONS/SIGNIFICANCE: Our case study has identified, and explained the functional relationships among, the critical features of the ED Program's approach to CE. It has also illuminated how these features were meaningful to stakeholders and contributed to garnering support within the host communities for the open-release trials. Our findings reveal how translating ethical intentions into effective action is more socially complex than is currently reflected in the CE literature. Because our case study delineates the critical features of the ED Program's approach to CE, it can serve as a framework for other programs to follow when designing their own strategies. And because the findings outline a theory of change for CE, it can also serve as a starting point for developing an evaluation framework for CE.}, } @article {pmid25870270, year = {2015}, author = {Perlman, SJ and Hodson, CN and Hamilton, PT and Opit, GP and Gowen, BE}, title = {Maternal transmission, sex ratio distortion, and mitochondria.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {112}, number = {33}, pages = {10162-10168}, pmid = {25870270}, issn = {1091-6490}, mesh = {Animals ; Arthropods/microbiology ; Bacteria/genetics ; Base Sequence ; Cell Nucleus/genetics ; DNA Barcoding, Taxonomic ; Electron Transport Complex IV/metabolism ; Female ; Haplotypes ; *Inheritance Patterns ; Insecta/microbiology ; Male ; Mitochondria/*genetics ; Molecular Sequence Data ; *Polymorphism, Genetic ; Sequence Analysis, DNA ; *Sex Ratio ; Symbiosis ; Wolbachia/physiology ; }, abstract = {In virtually all multicellular eukaryotes, mitochondria are transmitted exclusively through one parent, usually the mother. In this short review, we discuss some of the major consequences of uniparental transmission of mitochondria, including deleterious effects in males and selection for increased transmission through females. Many of these consequences, particularly sex ratio distortion, have well-studied parallels in other maternally transmitted genetic elements, such as bacterial endosymbionts of arthropods. We also discuss the consequences of linkage between mitochondria and other maternally transmitted genetic elements, including the role of cytonuclear incompatibilities in maintaining polymorphism. Finally, as a case study, we discuss a recently discovered maternally transmitted sex ratio distortion in an insect that is associated with extraordinarily divergent mitochondria.}, } @article {pmid25861561, year = {2015}, author = {Williams, LE and Wernegreen, JJ}, title = {Genome evolution in an ancient bacteria-ant symbiosis: parallel gene loss among Blochmannia spanning the origin of the ant tribe Camponotini.}, journal = {PeerJ}, volume = {3}, number = {}, pages = {e881}, pmid = {25861561}, issn = {2167-8359}, support = {R01 GM062626/GM/NIGMS NIH HHS/United States ; }, abstract = {Stable associations between bacterial endosymbionts and insect hosts provide opportunities to explore genome evolution in the context of established mutualisms and assess the roles of selection and genetic drift across host lineages and habitats. Blochmannia, obligate endosymbionts of ants of the tribe Camponotini, have coevolved with their ant hosts for ∼40 MY. To investigate early events in Blochmannia genome evolution across this ant host tribe, we sequenced Blochmannia from two divergent host lineages, Colobopsis obliquus and Polyrhachis turneri, and compared them with four published genomes from Blochmannia of Camponotus sensu stricto. Reconstructed gene content of the last common ancestor (LCA) of these six Blochmannia genomes is reduced (690 protein coding genes), consistent with rapid gene loss soon after establishment of the symbiosis. Differential gene loss among Blochmannia lineages has affected cellular functions and metabolic pathways, including DNA replication and repair, vitamin biosynthesis and membrane proteins. Blochmannia of P. turneri (i.e., B. turneri) encodes an intact DnaA chromosomal replication initiation protein, demonstrating that loss of dnaA was not essential for establishment of the symbiosis. Based on gene content, B. obliquus and B. turneri are unable to provision hosts with riboflavin. Of the six sequenced Blochmannia, B. obliquus is the earliest diverging lineage (i.e., the sister group of other Blochmannia sampled) and encodes the fewest protein-coding genes and the most pseudogenes. We identified 55 genes involved in parallel gene loss, including glutamine synthetase, which may participate in nitrogen recycling. Pathways for biosynthesis of coenzyme A, terpenoids and riboflavin were lost in multiple lineages, suggesting relaxed selection on the pathway after inactivation of one component. Analysis of Illumina read datasets did not detect evidence of plasmids encoding missing functions, nor the presence of coresident symbionts other than Wolbachia. Although gene order is strictly conserved in four Blochmannia of Camponotus sensu stricto, comparisons with deeply divergent lineages revealed inversions in eight genomic regions, indicating ongoing recombination despite ancestral loss of recA. In sum, the addition of two Blochmannia genomes of divergent host lineages enables reconstruction of early events in evolution of this symbiosis and suggests that Blochmannia lineages may experience distinct, host-associated selective pressures. Understanding how evolutionary forces shape genome reduction in this system may help to clarify forces driving gene loss in other bacteria, including intracellular pathogens.}, } @article {pmid25856898, year = {2014}, author = {Hong, SC and Lv, Y and Fang, FJ and Zhu, CL}, title = {[Presumptive role of Wolbachia in deltamethrin resistance of Culex pipiens pallens].}, journal = {Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control}, volume = {26}, number = {6}, pages = {672-4, 680}, pmid = {25856898}, issn = {1005-6661}, mesh = {Animals ; Base Sequence ; Culex/*drug effects/growth & development/*microbiology ; Female ; *Insecticide Resistance ; Insecticides/*pharmacology ; Male ; Molecular Sequence Data ; Nitriles/*pharmacology ; Pyrethrins/*pharmacology ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {OBJECTIVE: To investigate the relationship between Wolbachia and deltamethrin resistance in Culex pipiens pallens.

METHODS: PCR was used to detect Wolbachia in Culex pipiens pallens and qRT-PCR was performed to determine and compare the expression of Wolbachia between deltamethrin- resistant and - susceptible strains of Culex pipiens pallens.

RESULTS: Wolbachia was detected in Culex pipiens pallens in the laboratory. The expression of Wolbachia was 18.42, 3.69, 4.43, 3.96, 6.31, 1.55 and 3.76 folds higher in the deltamethrin-resistant strain than in susceptible strain in the egg, 1st, 2nd, 3rd, 4th stages, and male and female adults, but there was no statistical difference in the pupae stage. The expression of Wolbachia was 2.64 folds higher in deltamethrin-resistant females than in susceptible females which were caught in Jiangxinzhou of Nanjing.

CONCLUSION: Wolbachia is associated with deltamethrin resistance in Culex pipines pallens.}, } @article {pmid25852655, year = {2015}, author = {Chandler, JA and Liu, RM and Bennett, SN}, title = {RNA shotgun metagenomic sequencing of northern California (USA) mosquitoes uncovers viruses, bacteria, and fungi.}, journal = {Frontiers in microbiology}, volume = {6}, number = {}, pages = {185}, pmid = {25852655}, issn = {1664-302X}, abstract = {Mosquitoes, most often recognized for the microbial agents of disease they may carry, harbor diverse microbial communities that include viruses, bacteria, and fungi, collectively called the microbiota. The composition of the microbiota can directly and indirectly affect disease transmission through microbial interactions that could be revealed by its characterization in natural populations of mosquitoes. Furthermore, the use of shotgun metagenomic sequencing (SMS) approaches could allow the discovery of unknown members of the microbiota. In this study, we use RNA SMS to characterize the microbiota of seven individual mosquitoes (species include Culex pipiens, Culiseta incidens, and Ochlerotatus sierrensis) collected from a variety of habitats in California, USA. Sequencing was performed on the Illumina HiSeq platform and the resulting sequences were quality-checked and assembled into contigs using the A5 pipeline. Sequences related to single stranded RNA viruses of the Bunyaviridae and Rhabdoviridae were uncovered, along with an unclassified genus of double-stranded RNA viruses. Phylogenetic analysis finds that in all three cases, the closest relatives of the identified viral sequences are other mosquito-associated viruses, suggesting widespread host-group specificity among disparate viral taxa. Interestingly, we identified a Narnavirus of fungi, also reported elsewhere in mosquitoes, that potentially demonstrates a nested host-parasite association between virus, fungi, and mosquito. Sequences related to 8 bacterial families and 13 fungal families were found across the seven samples. Bacillus and Escherichia/Shigella were identified in all samples and Wolbachia was identified in all Cx. pipiens samples, while no single fungal genus was found in more than two samples. This study exemplifies the utility of RNA SMS in the characterization of the natural microbiota of mosquitoes and, in particular, the value of identifying all microbes associated with a specific host.}, } @article {pmid25849993, year = {2015}, author = {Kamalakannan, V and Shiny, A and Babu, S and Narayanan, RB}, title = {Autophagy protects monocytes from Wolbachia heat shock protein 60-induced apoptosis and senescence.}, journal = {PLoS neglected tropical diseases}, volume = {9}, number = {4}, pages = {e0003675}, pmid = {25849993}, issn = {1935-2735}, mesh = {Apoptosis/*drug effects/physiology ; *Autophagy ; Chaperonin 60/metabolism/*pharmacology ; Humans ; Inflammation ; Interleukin-6 ; Lymphocytes ; Male ; Membrane Potential, Mitochondrial ; Monocytes/*drug effects/physiology ; NF-kappa B/metabolism ; Protective Agents ; Reactive Oxygen Species/metabolism ; Signal Transduction ; Tumor Necrosis Factor-alpha ; Wolbachia/*metabolism ; }, abstract = {Monocyte dysfunction by filarial antigens has been a major mechanism underlying immune evasion following hyporesponsiveness during patent lymphatic filariasis. Recent studies have initiated a paradigm shift to comprehend the immunological interactions of Wolbachia and its antigens in inflammation, apoptosis, lymphocyte anergy, etc. Here we showed that recombinant Wolbachia heat shock protein 60 (rWmhsp60) interacts with TLR-4 and induces apoptosis in monocytes of endemic normal but not in chronic patients. Higher levels of reactive oxygen species (ROS) induced after TLR-4 stimulation resulted in loss of mitochondrial membrane potential and caspase cascade activation, which are the plausible reason for apoptosis. Furthermore, release in ROS owing to TLR-4 signaling resulted in the activation of NF-κB p65 nuclear translocation which leads to inflammation and apoptosis via TNF receptor pathway following the increase in IL-6 and TNF-α level. Here for the first time, we report that in addition to apoptosis, rWmhsp60 antigen in filarial pathogenesis also induces molecular senescence in monocytes. Targeting TLR-4, therefore, presents a promising candidate for treating rWmhsp60-induced apoptosis and senescence. Strikingly, induction of autophagy by rapamycin detains TLR-4 in late endosomes and subverts TLR-4-rWmhsp60 interaction, thus protecting TLR-4-mediated apoptosis and senescence. Furthermore, rapamycin-induced monocytes were unresponsive to rWmhsp60, and activated lymphocytes following PHA stimulation. This study demonstrates that autophagy mediates the degradation of TLR-4 signaling and protects monocytes from rWmhsp60 induced apoptosis and senescence.}, } @article {pmid25849812, year = {2015}, author = {Zhang, D and Zheng, X and Xi, Z and Bourtzis, K and Gilles, JR}, title = {Combining the sterile insect technique with the incompatible insect technique: I-impact of wolbachia infection on the fitness of triple- and double-infected strains of Aedes albopictus.}, journal = {PloS one}, volume = {10}, number = {4}, pages = {e0121126}, pmid = {25849812}, issn = {1932-6203}, mesh = {Aedes/growth & development/*microbiology ; Animals ; Female ; *Host-Pathogen Interactions ; Humans ; Insect Vectors/*microbiology ; Longevity ; Male ; *Symbiosis ; Wolbachia/*physiology ; Yellow Fever/microbiology/*transmission ; }, abstract = {The mosquito species Aedes albopictus is a major vector of the human diseases dengue and chikungunya. Due to the lack of efficient and sustainable methods to control this mosquito species, there is an increasing interest in developing and applying the sterile insect technique (SIT) and the incompatible insect technique (IIT), separately or in combination, as population suppression approaches. Ae. albopictus is naturally double-infected with two Wolbachia strains, wAlbA and wAlbB. A new triple Wolbachia-infected strain (i.e., a strain infected with wAlbA, wAlbB, and wPip), known as HC and expressing strong cytoplasmic incompatibility (CI) in appropriate matings, was recently developed. In the present study, we compared several fitness traits of three Ae. albopictus strains (triple-infected, double-infected and uninfected), all of which were of the same genetic background ("Guangzhou City, China") and were reared under the same conditions. Investigation of egg-hatching rate, survival of pupae and adults, sex ratio, duration of larval stages (development time from L1 to pupation), time to emergence (development time from L1 to adult emergence), wing length, female fecundity and adult longevity indicated that the presence of Wolbachia had only a minimal effect on host fitness. Based on this evidence, the HC strain is currently under consideration for mass rearing and application in a combined SIT-IIT strategy to control natural populations of Ae. albopictus in mainland China.}, } @article {pmid25845868, year = {2015}, author = {Shrivastava, N and Nag, JK and Pandey, J and Tripathi, RP and Shah, P and Siddiqi, MI and Misra-Bhattacharya, S}, title = {Homology modeling of NAD+-dependent DNA ligase of the Wolbachia endosymbiont of Brugia malayi and its drug target potential using dispiro-cycloalkanones.}, journal = {Antimicrobial agents and chemotherapy}, volume = {59}, number = {7}, pages = {3736-3747}, pmid = {25845868}, issn = {1098-6596}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Brugia malayi/*microbiology ; DNA Ligase ATP ; DNA Ligases/*antagonists & inhibitors/drug effects ; Escherichia coli/genetics/metabolism ; Filaricides/*pharmacology ; Gerbillinae ; Ketones/chemical synthesis/*pharmacology ; Male ; Microbial Sensitivity Tests ; Models, Molecular ; Molecular Docking Simulation ; Murinae/parasitology ; Spiro Compounds/chemical synthesis/*pharmacology ; Symbiosis ; Wolbachia/*drug effects/enzymology ; }, abstract = {Lymphatic filarial nematodes maintain a mutualistic relationship with the endosymbiont Wolbachia. Depletion of Wolbachia produces profound defects in nematode development, fertility, and viability and thus has great promise as a novel approach for treating filarial diseases. NAD(+)-dependent DNA ligase is an essential enzyme of DNA replication, repair, and recombination. Therefore, in the present study, the antifilarial drug target potential of the NAD(+)-dependent DNA ligase of the Wolbachia symbiont of Brugia malayi (wBm-LigA) was investigated using dispiro-cycloalkanone compounds. Dispiro-cycloalkanone specifically inhibited the nick-closing and cohesive-end ligation activities of the enzyme without inhibiting human or T4 DNA ligase. The mode of inhibition was competitive with the NAD(+) cofactor. Docking studies also revealed the interaction of these compounds with the active site of the target enzyme. The adverse effects of these inhibitors were observed on adult and microfilarial stages of B. malayi in vitro, and the most active compounds were further monitored in vivo in jirds and mastomys rodent models. Compounds 1, 2, and 5 had severe adverse effects in vitro on the motility of both adult worms and microfilariae at low concentrations. Compound 2 was the best inhibitor, with the lowest 50% inhibitory concentration (IC50) (1.02 μM), followed by compound 5 (IC50, 2.3 μM) and compound 1 (IC50, 2.9 μM). These compounds also exhibited the same adverse effect on adult worms and microfilariae in vivo (P < 0.05). These compounds also tremendously reduced the wolbachial load, as evident by quantitative real-time PCR (P < 0.05). wBm-LigA thus shows great promise as an antifilarial drug target, and dispiro-cycloalkanone compounds show great promise as antifilarial lead candidates.}, } @article {pmid25826386, year = {2015}, author = {Serbus, LR and White, PM and Silva, JP and Rabe, A and Teixeira, L and Albertson, R and Sullivan, W}, title = {The impact of host diet on Wolbachia titer in Drosophila.}, journal = {PLoS pathogens}, volume = {11}, number = {3}, pages = {e1004777}, pmid = {25826386}, issn = {1553-7374}, support = {T34 GM007910/GM/NIGMS NIH HHS/United States ; 2T34GM007910/GM/NIGMS NIH HHS/United States ; }, mesh = {*Animal Feed ; Animals ; Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster ; Female ; Oocytes/metabolism/*microbiology ; Sirolimus/pharmacology ; Symbiosis/drug effects/*physiology ; Transcription Factors/genetics/metabolism ; Wolbachia/cytology/*metabolism ; }, abstract = {While a number of studies have identified host factors that influence endosymbiont titer, little is known concerning environmental influences on titer. Here we examined nutrient impact on maternally transmitted Wolbachia endosymbionts in Drosophila. We demonstrate that Drosophila reared on sucrose- and yeast-enriched diets exhibit increased and reduced Wolbachia titers in oogenesis, respectively. The yeast-induced Wolbachia depletion is mediated in large part by the somatic TOR and insulin signaling pathways. Disrupting TORC1 with the small molecule rapamycin dramatically increases oocyte Wolbachia titer, whereas hyper-activating somatic TORC1 suppresses oocyte titer. Furthermore, genetic ablation of insulin-producing cells located in the Drosophila brain abolished the yeast impact on oocyte titer. Exposure to yeast-enriched diets altered Wolbachia nucleoid morphology in oogenesis. Furthermore, dietary yeast increased somatic Wolbachia titer overall, though not in the central nervous system. These findings highlight the interactions between Wolbachia and germline cells as strongly nutrient-sensitive, and implicate conserved host signaling pathways by which nutrients influence Wolbachia titer.}, } @article {pmid25813874, year = {2015}, author = {Cerveau, N and Gilbert, C and Liu, C and Garrett, RA and Grève, P and Bouchon, D and Cordaux, R}, title = {Genomic context drives transcription of insertion sequences in the bacterial endosymbiont Wolbachia wVulC.}, journal = {Gene}, volume = {564}, number = {1}, pages = {81-86}, doi = {10.1016/j.gene.2015.03.044}, pmid = {25813874}, issn = {1879-0038}, mesh = {Animals ; DNA Transposable Elements ; Gene Expression Regulation, Bacterial ; *Genome, Bacterial ; Isopoda/microbiology ; Promoter Regions, Genetic ; Symbiosis ; Transcription, Genetic ; Wolbachia/*genetics ; }, abstract = {Transposable elements (TEs) are DNA pieces that are present in almost all the living world at variable genomic density. Due to their mobility and density, TEs are involved in a large array of genomic modifications. In eukaryotes, TE expression has been studied in detail in several species. In prokaryotes, studies of IS expression are generally linked to particular copies that induce a modification of neighboring gene expression. Here we investigated global patterns of IS transcription in the Alphaproteobacterial endosymbiont Wolbachia wVulC, using both RT-PCR and bioinformatic analyses. We detected several transcriptional promoters in all IS groups. Nevertheless, only one of the potentially functional IS groups possesses a promoter located upstream of the transposase gene, that could lead up to the production of a functional protein. We found that the majority of IS groups are expressed whatever their functional status. RT-PCR analyses indicate that the transcription of two IS groups lacking internal promoters upstream of the transposase start codon may be driven by the genomic environment. We confirmed this observation with the transcription analysis of individual copies of one IS group. These results suggest that the genomic environment is important for IS expression and it could explain, at least partly, copy number variability of the various IS groups present in the wVulC genome and, more generally, in bacterial genomes.}, } @article {pmid25812130, year = {2015}, author = {Calvitti, M and Marini, F and Desiderio, A and Puggioli, A and Moretti, R}, title = {Wolbachia density and cytoplasmic incompatibility in Aedes albopictus: concerns with using artificial Wolbachia infection as a vector suppression tool.}, journal = {PloS one}, volume = {10}, number = {3}, pages = {e0121813}, pmid = {25812130}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Animals ; Bacterial Load ; Female ; Insect Vectors/microbiology ; Male ; Symbiosis ; *Wolbachia/classification/genetics ; }, abstract = {The mosquito Aedes albopictusi is a competent vector of harmful human pathogens, including viruses causing dengue and chikungunya. Cytoplasmic incompatibility (CI) induced by endosymbiotic Wolbachia can be used to produce functionally sterile males that can be released in the field as a suppression tool against this mosquito. Because the available sexing methods are not efficient enough to avoid unintentional release of a few transinfected females, we assessed the CI pattern in crosses between wPip Wolbachia-transinfected (ARwP) females and wild-type males of Ae. albopictus in this study. Quantitative polymerase chain reaction was used to monitor the titer of the Wolbachia strains that naturally infect Ae. albopictus, that is, wAlbA and wAlbB, in age-controlled males and females. Data were coupled with incompatibility level detected when the above-mentioned males were crossed with ARwP females. Wolbachia infection titer was also monitored in samples of wild caught males. Incompatibility level was positively correlated only with wAlbA density. Crosses between wild-type males having very low wAlbA density (<0.001 wAlbA/actin copy numbers) and ARwP females were partially fertile (CIcorr = 68.06 ± 6.20). Individuals with low wAlbA titer were frequently found among sampled wild males (30%-50% depending on the site and period). ARwP males can be as considered as a very promising tool for suppressing Ae. albopictus. However, crosses between wild males having low wAlbA density and ARwP females may be partially fertile. In the case of local establishment of the transinfected mosquito line, this occurrence may favor the replacement of the wild-type mosquitoes with the ARwP line, thus reducing the long-term efficacy of incompatible insect technique. Various alternative strategies have been discussed to prevent this risk and to exploit Wolbachia as a tool to control Ae. albopictus.}, } @article {pmid25809075, year = {2015}, author = {Christensen, S and Serbus, LR}, title = {Comparative analysis of wolbachia genomes reveals streamlining and divergence of minimalist two-component systems.}, journal = {G3 (Bethesda, Md.)}, volume = {5}, number = {5}, pages = {983-996}, pmid = {25809075}, issn = {2160-1836}, mesh = {Amino Acid Sequence ; Amino Acids ; Computational Biology/methods ; Gene Order ; Genes, Bacterial ; Genetic Loci ; *Genome, Bacterial ; *Genomics/methods ; Molecular Sequence Data ; Open Reading Frames ; Operon ; Protein Interaction Domains and Motifs ; Sequence Alignment ; *Signal Transduction ; Wolbachia/*genetics/*metabolism ; }, abstract = {Two-component regulatory systems are commonly used by bacteria to coordinate intracellular responses with environmental cues. These systems are composed of functional protein pairs consisting of a sensor histidine kinase and cognate response regulator. In contrast to the well-studied Caulobacter crescentus system, which carries dozens of these pairs, the streamlined bacterial endosymbiont Wolbachia pipientis encodes only two pairs: CckA/CtrA and PleC/PleD. Here, we used bioinformatic tools to compare characterized two-component system relays from C. crescentus, the related Anaplasmataceae species Anaplasma phagocytophilum and Ehrlichia chaffeensis, and 12 sequenced Wolbachia strains. We found the core protein pairs and a subset of interacting partners to be highly conserved within Wolbachia and these other Anaplasmataceae. Genes involved in two-component signaling were positioned differently within the various Wolbachia genomes, whereas the local context of each gene was conserved. Unlike Anaplasma and Ehrlichia, Wolbachia two-component genes were more consistently found clustered with metabolic genes. The domain architecture and key functional residues standard for two-component system proteins were well-conserved in Wolbachia, although residues that specify cognate pairing diverged substantially from other Anaplasmataceae. These findings indicate that Wolbachia two-component signaling pairs share considerable functional overlap with other α-proteobacterial systems, whereas their divergence suggests the potential for regulatory differences and cross-talk.}, } @article {pmid25807173, year = {2015}, author = {Gauthier, JP and Outreman, Y and Mieuzet, L and Simon, JC}, title = {Bacterial communities associated with host-adapted populations of pea aphids revealed by deep sequencing of 16S ribosomal DNA.}, journal = {PloS one}, volume = {10}, number = {3}, pages = {e0120664}, pmid = {25807173}, issn = {1932-6203}, mesh = {Animals ; Aphids/classification/*genetics/microbiology ; Bacteria/isolation & purification ; Buchnera/isolation & purification ; Cluster Analysis ; Erwinia/isolation & purification ; Genetic Variation ; Genotype ; High-Throughput Nucleotide Sequencing ; Molecular Sequence Data ; Pantoea/isolation & purification ; Phylogeny ; RNA, Ribosomal, 16S ; Sequence Analysis, RNA ; Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {Associations between microbes and animals are ubiquitous and hosts may benefit from harbouring microbial communities through improved resource exploitation or resistance to environmental stress. The pea aphid, Acyrthosiphon pisum, is the host of heritable bacterial symbionts, including the obligate endosymbiont Buchnera aphidicola and several facultative symbionts. While obligate symbionts supply aphids with key nutrients, facultative symbionts influence their hosts in many ways such as protection against natural enemies, heat tolerance, color change and reproduction alteration. The pea aphid also encompasses multiple plant-specialized biotypes, each adapted to one or a few legume species. Facultative symbiont communities differ strongly between biotypes, although bacterial involvement in plant specialization is uncertain. Here, we analyse the diversity of bacterial communities associated with nine biotypes of the pea aphid complex using amplicon pyrosequencing of 16S rRNA genes. Combined clustering and phylogenetic analyses of 16S sequences allowed identifying 21 bacterial OTUs (Operational Taxonomic Unit). More than 98% of the sequencing reads were assigned to known pea aphid symbionts. The presence of Wolbachia was confirmed in A. pisum while Erwinia and Pantoea, two gut associates, were detected in multiple samples. The diversity of bacterial communities harboured by pea aphid biotypes was very low, ranging from 3 to 11 OTUs across samples. Bacterial communities differed more between than within biotypes but this difference did not correlate with the genetic divergence between biotypes. Altogether, these results confirm that the aphid microbiota is dominated by a few heritable symbionts and that plant specialization is an important structuring factor of bacterial communities associated with the pea aphid complex. However, since we examined the microbiota of aphid samples kept a few generations in controlled conditions, it may be that bacterial diversity was underestimated due to the possible loss of environmental or transient taxa.}, } @article {pmid25801227, year = {2015}, author = {Méndez, JC and Carretón, E and Martínez-Subiela, S and Tvarijonaviciute, A and Cerón, JJ and Montoya-Alonso, JA}, title = {Acute phase protein response in heartworm-infected dogs after adulticide treatment.}, journal = {Veterinary parasitology}, volume = {209}, number = {3-4}, pages = {197-201}, doi = {10.1016/j.vetpar.2015.02.036}, pmid = {25801227}, issn = {1873-2550}, mesh = {Acute-Phase Proteins/*metabolism ; Animals ; Anthelmintics/administration & dosage/therapeutic use ; Dirofilariasis/*blood/drug therapy/metabolism ; Dog Diseases/blood/drug therapy/*parasitology ; Dogs ; Doxycycline/administration & dosage/*therapeutic use ; Drug Therapy, Combination ; Female ; Ivermectin/administration & dosage/*therapeutic use ; Male ; Time Factors ; }, abstract = {During the adulticide treatment of Dirofilaria immitis the worms die releasing fragments of parasites and causing pulmonary thromboembolisms which could exacerbate the clinical condition. To determine the utility of acute phase proteins (APPs) to monitor the progression of the treatment, different positive [C-reactive protein (CRP), haptoglobin (hp)] and negative [albumin, paraoxonase-1(PON-1)] APPs were measured in 15 heartworm-infected dogs (5 with high and 10 with low parasite burden) following adulticide treatment. The results showed increased concentrations of CRP, decreased concentrations of haptoglobin and PON-1 in infected dogs before starting the treatment. Progressive but not significant increases were observed in PON-1 activity and albumin concentration along the treatment. After the treatment with doxycycline and ivermectine a decrease in CRP and Hp levels was experienced, which could reflect a reduction of the vascular inflammation caused by the elimination of Wolbachia and reduction of microfilariae. Fifteen days after the first melarsomine injection, marked increases in CRP and Hp were observed, which could be due to pulmonary inflammation and thromboembolism caused by the post-adulticide death of the worms. This increase was greater in dogs with high parasite burden. As the pathology disappeared, there was an improvement in the concentrations of CRP and Hp, returning into reference values in dogs with low parasite burden at the end of the treatment. The measurement of CRP and Hp could be a resource of support to evaluate the magnitude of the post-adulticide complications during the adulticide treatment of D. immitis.}, } @article {pmid25787763, year = {2015}, author = {Ferguson, NM and Kien, DT and Clapham, H and Aguas, R and Trung, VT and Chau, TN and Popovici, J and Ryan, PA and O'Neill, SL and McGraw, EA and Long, VT and Dui, le T and Nguyen, HL and Chau, NV and Wills, B and Simmons, CP}, title = {Modeling the impact on virus transmission of Wolbachia-mediated blocking of dengue virus infection of Aedes aegypti.}, journal = {Science translational medicine}, volume = {7}, number = {279}, pages = {279ra37}, pmid = {25787763}, issn = {1946-6242}, support = {/WT_/Wellcome Trust/United Kingdom ; 089276/WT_/Wellcome Trust/United Kingdom ; MR/K010174/1/MRC_/Medical Research Council/United Kingdom ; U01 GM110721/GM/NIGMS NIH HHS/United States ; }, mesh = {Adolescent ; Adult ; Aedes/*microbiology/*virology ; Animals ; Dengue/*microbiology/transmission/*virology ; Dengue Virus/*physiology ; Disease Vectors ; Female ; Humans ; Male ; Models, Theoretical ; Prospective Studies ; Saliva/microbiology/virology ; Wolbachia/*physiology ; Young Adult ; }, abstract = {Dengue is the most common arboviral infection of humans and is a public health burden in more than 100 countries. Aedes aegypti mosquitoes stably infected with strains of the intracellular bacterium Wolbachia are resistant to dengue virus (DENV) infection and are being tested in field trials. To mimic field conditions, we experimentally assessed the vector competence of A. aegypti carrying the Wolbachia strains wMel and wMelPop after challenge with viremic blood from dengue patients. We found that wMelPop conferred strong resistance to DENV infection of mosquito abdomen tissue and largely prevented disseminated infection. wMel conferred less resistance to infection of mosquito abdomen tissue, but it did reduce the prevalence of mosquitoes with infectious saliva. A mathematical model of DENV transmission incorporating the dynamics of viral infection in humans and mosquitoes was fitted to the data collected. Model predictions suggested that wMel would reduce the basic reproduction number, R0, of DENV transmission by 66 to 75%. Our results suggest that establishment of wMelPop-infected A. aegypti at a high frequency in a dengue-endemic setting would result in the complete abatement of DENV transmission. Establishment of wMel-infected A. aegypti is also predicted to have a substantial effect on transmission that would be sufficient to eliminate dengue in low or moderate transmission settings but may be insufficient to achieve complete control in settings where R0 is high. These findings develop a framework for selecting Wolbachia strains for field releases and for calculating their likely impact.}, } @article {pmid25772521, year = {2015}, author = {de Oliveira, CD and Gonçalves, DS and Baton, LA and Shimabukuro, PH and Carvalho, FD and Moreira, LA}, title = {Broader prevalence of Wolbachia in insects including potential human disease vectors.}, journal = {Bulletin of entomological research}, volume = {105}, number = {3}, pages = {305-315}, doi = {10.1017/S0007485315000085}, pmid = {25772521}, issn = {1475-2670}, mesh = {*Animal Distribution ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Brazil ; Cytoskeletal Proteins/genetics ; High-Throughput Screening Assays/methods ; Insect Vectors/*microbiology ; Insecta/*microbiology/physiology ; RNA, Ribosomal, 16S/genetics ; Real-Time Polymerase Chain Reaction/methods ; Species Specificity ; Wolbachia/*genetics ; }, abstract = {Wolbachia are intracellular, maternally transmitted bacteria considered the most abundant endosymbionts found in arthropods. They reproductively manipulate their host in order to increase their chances of being transmitted to the offspring, and currently are being used as a tool to control vector-borne diseases. Studies on distribution of Wolbachia among its arthropod hosts are important both for better understanding why this bacterium is so common, as well as for its potential use as a biological control agent. Here, we studied the incidence of Wolbachia in a broad range of insect species, collected from different regions of Brazil, using three genetic markers (16S rRNA, wsp and ftsZ), which varied in terms of their sensitivity to detect this bacterium. The overall incidence of Wolbachia among species belonging to 58 families and 14 orders was 61.9%. The most common positive insect orders were Coleoptera, Diptera, Hemiptera and Hymenoptera, with Diptera and Hemiptera having the highest numbers of Wolbachia-positive families. They included potential human disease vectors whose infection status has never been reported before. Our study further shows the importance of using quantitative polymerase chain reaction for high-throughput and sensitive Wolbachia screening.}, } @article {pmid25769472, year = {2015}, author = {Menozzi, A and Bertini, S and Turin, L and Serventi, P and Kramer, L and Bazzocchi, C}, title = {Doxycycline levels and anti-Wolbachia antibodies in sera from dogs experimentally infected with Dirofilaria immitis and treated with a combination of ivermectin/doxycycline.}, journal = {Veterinary parasitology}, volume = {209}, number = {3-4}, pages = {281-284}, doi = {10.1016/j.vetpar.2015.02.023}, pmid = {25769472}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/blood/therapeutic use ; Antibodies, Bacterial/*blood ; Antiparasitic Agents/administration & dosage/therapeutic use ; Dirofilariasis/*drug therapy ; Dog Diseases/drug therapy/immunology/*parasitology ; Dogs ; Doxycycline/administration & dosage/*blood/therapeutic use ; Drug Therapy, Combination ; Enzyme-Linked Immunosorbent Assay ; Female ; Ivermectin/administration & dosage/*therapeutic use ; Male ; Wolbachia/*immunology ; }, abstract = {Sera from Dirofilaria immitis-experimentally infected dogs treated with a combination of ivermectin/doxycycline were analysed for doxycycline levels by HPLC and anti-Wolbachia Surface Protein (rWSP) antibodies by ELISA and compared with sera from dogs treated with doxycycline alone. Results show that doxycycline levels were not statistically different between the two groups. Circulating anti-WSP antibody titres were markedly lower in both treatment groups when compared to control D. immitis infected dogs, indicating that doxycycline is able to reduce Wolbachia and prevent the immune response against the bacteria. The combination treatment protocol has been shown to be highly adulticidal and further studies are needed to better understand the interaction between doxycycline and ivermectin in D. immitis infected dogs.}, } @article {pmid25768841, year = {2015}, author = {Atyame, CM and Cattel, J and Lebon, C and Flores, O and Dehecq, JS and Weill, M and Gouagna, LC and Tortosa, P}, title = {Wolbachia-based population control strategy targeting Culex quinquefasciatus mosquitoes proves efficient under semi-field conditions.}, journal = {PloS one}, volume = {10}, number = {3}, pages = {e0119288}, pmid = {25768841}, issn = {1932-6203}, mesh = {Animals ; Culex/microbiology/*physiology ; Culicidae/microbiology/*physiology ; Cytoplasm/microbiology/physiology ; Female ; Indian Ocean ; Male ; Mosquito Control/methods ; Population Control/methods ; Reproduction/physiology ; Wolbachia/*genetics ; }, abstract = {In mosquitoes, the maternally inherited bacterial Wolbachia induce a form of embryonic lethality called cytoplasmic incompatibility (CI). This property can be used to reduce the density of mosquito field populations through inundative releases of incompatible males in order to sterilize females (Incompatible Insect Technique, or IIT, strategy). We have previously constructed the LR[wPip(Is)] line representing a good candidate for controlling field populations of the Culex quinquefasciatus mosquito in the islands of the south-western Indian Ocean. The main purpose of the present study was to fill the gap between laboratory experiments and field implementation, i.e. assessing mating competitiveness of these incompatible males under semi-field conditions. In a first set of experiments, we analyzed crossing relationships between LR[wPip(Is)] males and La Réunion field females collected as larvae in 19 distinct localities throughout the island. This investigation revealed total embryonic mortality, confirming the strong sterilizing capacity of LR[wPip(Is)] males. Subsequently, mating competitiveness of LR[wPip(Is)] males was assessed under semi-field conditions in the presence of field males and females from La Réunion. Confrontations were carried out in April and December using different ratios of LR[wPip(Is)] to field males. The results indicated that the LR[wPip(Is)] males successfully compete with field males in mating with field females, displaying even higher competitiveness than field males in April. Our results support the implementation of small-scale field tests in order to assess the feasibility of IIT against Cx. quinquefasciatus in the islands of southwestern Indian Ocean where this mosquito species is a proven competent vector for human pathogens.}, } @article {pmid25764472, year = {2015}, author = {Zimmermann, BL and Bouchon, D and Almerão, MP and Araujo, PB}, title = {Wolbachia in Neotropical terrestrial isopods.}, journal = {FEMS microbiology ecology}, volume = {91}, number = {4}, pages = {}, doi = {10.1093/femsec/fiv025}, pmid = {25764472}, issn = {1574-6941}, mesh = {Animals ; Bacterial Typing Techniques ; Base Sequence ; Biodiversity ; Biological Evolution ; DNA, Bacterial/genetics ; Genetic Variation/genetics ; Geography ; Isopoda/*microbiology ; Multilocus Sequence Typing ; Phylogeny ; Sequence Analysis, DNA ; South America ; Symbiosis ; Wolbachia/*classification/*genetics ; }, abstract = {Despite Wolbachia being widespread among terrestrial isopods, studies on this symbiotic relationship are still incipient in the Neotropical region. The aims of the present study were to investigate the presence and prevalence of Wolbachia in natural populations of terrestrial isopod species in South America, and to analyze the diversity and phylogenetic relationships of Wolbachia strains. A total of 1172 individuals representing 11 families and 35 species were analyzed. We observed distinct evolutionary scenarios according to the geographical origins of the species: strains harbored by most of the introduced species belong to the Oniclade in supergroup B and are identical to those found in their original ecozone (i.e. Palearctic). On the other hand, the strains found in native Neotropical terrestrial isopods showed low prevalence, high diversity and none of them belonged to the Oniclade, although most belonged to supergroup B. The dynamics of infection in Neotropical species seems to be the result of several events of loss and acquisition of the bacteria, which refutes the hypothesis of an ancestral acquisition of Wolbachia in Oniscidea. The presence of strains from supergroups A and F was also detected for the first time in terrestrial isopods, revealing a Wolbachia diversity previously unknown for this group of host.}, } @article {pmid25739293, year = {2014}, author = {Shaĭkevich, EV and Zakharov, IA}, title = {[Coevolution of symbiotic bacteria Wolbachia and host mtDNA in Russian populations of the Culex pipiens mosquito complex].}, journal = {Genetika}, volume = {50}, number = {11}, pages = {1390-1393}, pmid = {25739293}, issn = {0016-6758}, mesh = {Animals ; Culex/*genetics ; DNA, Mitochondrial/*genetics ; *Evolution, Molecular ; *Phylogeny ; Russia ; Symbiosis/*genetics ; Wolbachia/*genetics ; }, abstract = {The mitochondrial DNA phylogenies of closely related forms of mosquitoes from the Culex pipiens complex and of strains of the endosymbiotic bacteria Wolbachia pipientis were compared. Based on the cytochrome oxidase subunit I gene polymorphism, six mitochondrial haplotypes and four W. pipientis groups were discovered in mosquitoes from geographically remote populations. A strict correlation between the COI type and the type of W. pipientis proves the stable coinheritance and distribution of both cytoplasmic components in the examined mosquito populations and suggests either the absence or rarity of horizontal transfer of the symbionts in the Culex pipiens complex.}, } @article {pmid25739023, year = {2015}, author = {Woolfit, M and Algama, M and Keith, JM and McGraw, EA and Popovici, J}, title = {Discovery of putative small non-coding RNAs from the obligate intracellular bacterium Wolbachia pipientis.}, journal = {PloS one}, volume = {10}, number = {3}, pages = {e0118595}, pmid = {25739023}, issn = {1932-6203}, mesh = {Animals ; Computational Biology ; Conserved Sequence ; Drosophila melanogaster/microbiology ; Female ; Host Specificity ; Intracellular Space/*microbiology ; Male ; Organ Specificity ; RNA, Messenger/genetics/metabolism ; RNA, Small Untranslated/*genetics ; Sequence Analysis, RNA ; Transcription, Genetic ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia pipientis is an endosymbiotic bacterium that induces a wide range of effects in its insect hosts, including manipulation of reproduction and protection against pathogens. Little is known of the molecular mechanisms underlying the insect-Wolbachia interaction, though it is likely to be mediated via the secretion of proteins or other factors. There is an increasing amount of evidence that bacteria regulate many cellular processes, including secretion of virulence factors, using small non-coding RNAs (sRNAs), but sRNAs have not previously been described from Wolbachia. We have used two independent approaches, one based on comparative genomics and the other using RNA-Seq data generated for gene expression studies, to identify candidate sRNAs in Wolbachia. We experimentally characterized the expression of one of these candidates in four Wolbachia strains, and showed that it is differentially regulated in different host tissues and sexes. Given the roles played by sRNAs in other host-associated bacteria, the conservation of the candidate sRNAs between different Wolbachia strains, and the sex- and tissue-specific differential regulation we have identified, we hypothesise that sRNAs may play a significant role in the biology of Wolbachia, and in particular in its interactions with its host.}, } @article {pmid25717372, year = {2015}, author = {Bergman, CM and Haddrill, PR}, title = {Strain-specific and pooled genome sequences for populations of Drosophila melanogaster from three continents.}, journal = {F1000Research}, volume = {4}, number = {}, pages = {31}, pmid = {25717372}, issn = {2046-1402}, abstract = {To contribute to our general understanding of the evolutionary forces that shape variation in genome sequences in nature, we have sequenced genomes from 50 isofemale lines and six pooled samples from populations of Drosophila melanogaster on three continents. Analysis of raw and reference-mapped reads indicates the quality of these genomic sequence data is very high. Comparison of the predicted and experimentally-determined Wolbachia infection status of these samples suggests that strain or sample swaps are unlikely to have occurred in the generation of these data. Genome sequences are freely available in the European Nucleotide Archive under accession ERP009059. Isofemale lines can be obtained from the Drosophila Species Stock Center.}, } @article {pmid25710364, year = {2015}, author = {Wong, ZS and Brownlie, JC and Johnson, KN}, title = {Oxidative stress correlates with Wolbachia-mediated antiviral protection in Wolbachia-Drosophila associations.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {9}, pages = {3001-3005}, pmid = {25710364}, issn = {1098-5336}, mesh = {Animals ; Drosophila/immunology/*microbiology/*physiology ; Hydrogen Peroxide/analysis ; *Oxidative Stress ; Survival Analysis ; Viruses/*immunology ; Wolbachia/*growth & development ; }, abstract = {Wolbachia mediates antiviral protection in insect hosts and is being developed as a potential biocontrol agent to reduce the spread of insect-vectored viruses. Definition of the molecular mechanism that generates protection is important for understanding the tripartite interaction between host insect, Wolbachia, and virus. Elevated oxidative stress was previously reported for a mosquito line experimentally infected with Wolbachia, suggesting that oxidative stress is important for Wolbachia-mediated antiviral protection. However, Wolbachia experimentally introduced into mosquitoes impacts a range of host fitness traits, some of which are unrelated to antiviral protection. To explore whether elevated oxidative stress is associated with antiviral protection in Wolbachia-infected insects, we analyzed oxidative stress of five Wolbachia-infected Drosophila lines. In flies infected with protective Wolbachia strains, hydrogen peroxide concentrations were 1.25- to 2-fold higher than those in paired fly lines cured of Wolbachia infection. In contrast, there was no difference in the hydrogen peroxide concentrations in flies infected with nonprotective Wolbachia strains compared to flies cured of Wolbachia infection. Using a Drosophila mutant that produces increased levels of hydrogen peroxide, we investigated whether flies with high levels of endogenous reactive oxygen species had altered responses to virus infection and found that flies with high levels of endogenous hydrogen peroxide were less susceptible to virus-induced mortality. Taken together, these results suggest that elevated oxidative stress correlates with Wolbachia-mediated antiviral protection in natural Drosophila hosts.}, } @article {pmid25706947, year = {2015}, author = {Vanthournout, B and Hendrickx, F}, title = {Endosymbiont dominated bacterial communities in a dwarf spider.}, journal = {PloS one}, volume = {10}, number = {2}, pages = {e0117297}, pmid = {25706947}, issn = {1932-6203}, mesh = {Animals ; Bacteroidetes/genetics/*isolation & purification ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/*isolation & purification ; Spiders/*microbiology ; Symbiosis/*genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The microbial community of spiders is little known, with previous studies focussing primarily on the medical importance of spiders as vectors of pathogenic bacteria and on the screening of known cytoplasmic endosymbiont bacteria. These screening studies have been performed by means of specific primers that only amplify a selective set of endosymbionts, hampering the detection of unreported species in spiders. In order to have a more complete overview of the bacterial species that can be present in spiders, we applied a combination of a cloning assay, DGGE profiling and high-throughput sequencing on multiple individuals of the dwarf spider Oedothorax gibbosus. This revealed a co-infection of at least three known (Wolbachia, Rickettsia and Cardinium) and the detection of a previously unreported endosymbiont bacterium (Rhabdochlamydia) in spiders. 16S rRNA gene sequences of Rhabdochlamydia matched closely with those of Candidatus R. porcellionis, which is currently only reported as a pathogen from a woodlouse and with Candidatus R. crassificans reported from a cockroach. Remarkably, this bacterium appears to present in very high proportions in one of the two populations only, with all investigated females being infected. We also recovered Acinetobacter in high abundance in one individual. In total, more than 99% of approximately 4.5M high-throughput sequencing reads were restricted to these five bacterial species. In contrast to previously reported screening studies of terrestrial arthropods, our results suggest that the bacterial communities in this spider species are dominated by, or even restricted to endosymbiont bacteria. Given the high prevalence of endosymbiont species in spiders, this bacterial community pattern could be widespread in the Araneae order.}, } @article {pmid25695760, year = {2015}, author = {Rugman-Jones, PF and Seybold, SJ and Graves, AD and Stouthamer, R}, title = {Phylogeography of the walnut twig beetle, Pityophthorus juglandis, the vector of thousand cankers disease in North American walnut trees.}, journal = {PloS one}, volume = {10}, number = {2}, pages = {e0118264}, pmid = {25695760}, issn = {1932-6203}, mesh = {Animals ; Base Sequence ; Coleoptera/*genetics ; DNA, Mitochondrial/genetics ; Genes, Insect ; Molecular Sequence Data ; Phylogeography ; *Polymorphism, Restriction Fragment Length ; RNA, Ribosomal/genetics ; }, abstract = {Thousand cankers disease (TCD) of walnut trees (Juglans spp.) results from aggressive feeding in the phloem by the walnut twig beetle (WTB), Pityophthorus juglandis, accompanied by inoculation of its galleries with a pathogenic fungus, Geosmithia morbida. In 1960, WTB was only known from four U.S. counties (in Arizona, California, and New Mexico), but the species has now (2014) invaded over 115 counties, representing much of the western USA, and at least six states in the eastern USA. The eastern expansion places TCD in direct proximity to highly valuable (> $500 billion) native timber stands of eastern black walnut, Juglans nigra. Using mitochondrial DNA sequences, from nearly 1100 individuals, we examined variation among 77 samples of WTB populations across its extended range in the USA, revealing high levels of polymorphism and evidence of two divergent lineages. The highest level of genetic diversity for the different lineages was found in the neighboring Madrean Sky Island and Western New Mexico regions, respectively. Despite their proximity, there was little evidence of mixing between these regions, with only a single migrant detected among 179 beetles tested. Indeed, geographic overlap of the two lineages was only common in parts of Colorado and Utah. Just two haplotypes, from the same lineage, predominated over the vast majority of the recently expanded range. Tests for Wolbachia proved negative suggesting it plays no role in "driving" the spread of particular haplotypes, or in maintaining deep levels of intraspecific divergence in WTB. Genotyping of ribosomal RNA corroborated the mitochondrial lineages, but also revealed evidence of hybridization between them. Hybridization was particularly prevalent in the sympatric areas, also apparent in all invaded areas, but absent from the most haplotype-rich area of each mitochondrial lineage. Hypotheses about the specific status of WTB, its recent expansion, and potential evolutionary origins of TCD are discussed.}, } @article {pmid25692519, year = {2015}, author = {Dobson, AJ and Chaston, JM and Newell, PD and Donahue, L and Hermann, SL and Sannino, DR and Westmiller, S and Wong, AC and Clark, AG and Lazzaro, BP and Douglas, AE}, title = {Host genetic determinants of microbiota-dependent nutrition revealed by genome-wide analysis of Drosophila melanogaster.}, journal = {Nature communications}, volume = {6}, number = {}, pages = {6312}, pmid = {25692519}, issn = {2041-1723}, support = {R01 AI064950/AI/NIAID NIH HHS/United States ; R01 AI083932/AI/NIAID NIH HHS/United States ; R01 GM095372/GM/NIGMS NIH HHS/United States ; 1R01GM095372/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/*microbiology ; Gastrointestinal Tract/*microbiology ; Genetic Association Studies ; Genetic Variation ; Genotype ; Glucose/chemistry ; Glycogen/chemistry ; Lipids/chemistry ; *Microbiota ; Mutation ; *Nutritional Physiological Phenomena ; Phenotype ; Polymorphism, Single Nucleotide ; Signal Transduction ; Wolbachia/*physiology ; }, abstract = {Animals bear communities of gut microorganisms with substantial effects on animal nutrition, but the host genetic basis of these effects is unknown. Here we use Drosophila to demonstrate substantial among-genotype variation in the effects of eliminating the gut microbiota on five host nutritional indices (weight, protein, lipid, glucose and glycogen contents); this includes variation in both the magnitude and direction of microbiota-dependent effects. Genome-wide association studies to identify the genetic basis of the microbiota-dependent variation reveal polymorphisms in largely non-overlapping sets of genes associated with variation in the nutritional traits, including strong representation of conserved genes functioning in signalling. Key genes identified by the GWA study are validated by loss-of-function mutations that altered microbiota-dependent nutritional effects. We conclude that the microbiota interacts with the animal at multiple points in the signalling and regulatory networks that determine animal nutrition. These interactions with the microbiota are probably conserved across animals, including humans.}, } @article {pmid25686301, year = {2014}, author = {Clarke, LL and Ballweber, LR and Allen, K and Little, SE and Lappin, MR}, title = {Prevalence of select vector-borne disease agents in owned dogs of Ghana.}, journal = {Journal of the South African Veterinary Association}, volume = {85}, number = {1}, pages = {996}, doi = {10.4102/jsava.v85i1.996}, pmid = {25686301}, issn = {2224-9435}, mesh = {Animals ; Antibodies, Bacterial/*blood ; Antigens, Helminth ; DNA, Bacterial/isolation & purification ; Dirofilaria immitis ; *Disease Vectors ; Dog Diseases/*blood/epidemiology/immunology ; Dogs ; Ghana/epidemiology ; Ownership ; Ticks/microbiology/parasitology ; }, abstract = {Ticks, sera and ethylenediaminetetraacetic acid (EDTA) blood were collected from dogs evaluated at the Amakom Veterinary Clinic in Kumasi, Ghana. Sera were evaluated for Dirofilaria immitis antigen and antibodies against Borrelia burgdorferi, Anaplasma phagocytophilum and Ehrlichia canis. Conventional polymerase chain reaction assays designed to amplify the deoxyribonucleic acid (DNA) of Ehrlichia spp. or Anaplasma spp. or Neorickettsia spp. or Wolbachia spp., Babesia spp., Rickettsia spp., Hepatozoon spp., Bartonella spp. and the haemoplasmas were performed on DNA extracted from EDTA blood and all positive amplicons were sequenced. This small survey shows that the following vector-borne pathogens are present in urban Ghanian dogs: Ehrlichia canis, Hepatozoon canis,Dirofilaria immitis and Anaplasma platys. Bartonella henselae was isolated from ticks but not from the dogs.}, } @article {pmid25683566, year = {2015}, author = {Hu, HY and Li, ZX}, title = {A novel Wolbachia strain from the rice moth Corcyra cephalonica induces reproductive incompatibility in the whitefly Bemisia tabaci: sequence typing combined with phenotypic evidence.}, journal = {Environmental microbiology reports}, volume = {7}, number = {3}, pages = {508-515}, doi = {10.1111/1758-2229.12279}, pmid = {25683566}, issn = {1758-2229}, mesh = {Animals ; DNA, Bacterial/genetics ; Hemiptera/*microbiology/*physiology ; Moths/*microbiology ; Multilocus Sequence Typing ; *Reproduction ; Wolbachia/classification/genetics/*growth & development/isolation & purification ; }, abstract = {Wolbachia are a group of maternally inherited bacteria frequently found in arthropods and filarial nematodes. They have recently attracted attention for their ecological roles in manipulating host reproduction, their potential use in biological control of pest insects and medical significance. Classification of Wolbachia strains is currently solely based on molecular methods. However, the strains even with identical sequence types may induce different host phenotypes. Here we isolated a Wolbachia strain from the rice moth Corcyra cephalonica (designated as wCcep_B_BJ), which was shown to share multilocus sequence typing and Wolbachia surface protein hypervariable region profiles with a cytoplasmic incompatibility (CI)-inducing strain in supergroup B, but the phenotype wCcep_B_BJ may induce needs to be determined. We thus transinfected it into the whitefly Bemisia tabaci harbouring an A-Wolbachia through nymphal microinjection. Fluorescent in situ hybridization demonstrated that wCcep_B_BJ was successfully transinfected into B. tabaci and transmitted to offspring through host eggs. Reciprocal cross showed that wCcep_B_BJ induced a strong bidirectional CI in the transinfected host without imposing a significant cost on female fecundity. Our results suggest that wCcep_B_BJ may be a promising strain for biocontrol of B. tabaci, an important agricultural pest insect.}, } @article {pmid25681033, year = {2015}, author = {Lis, A and Maryańska-Nadachowska, A and Kajtoch, Ł}, title = {Relations of Wolbachia Infection with Phylogeography of Philaenus spumarius (Hemiptera: Aphrophoridae) Populations Within and Beyond the Carpathian Contact Zone.}, journal = {Microbial ecology}, volume = {70}, number = {2}, pages = {509-521}, pmid = {25681033}, issn = {1432-184X}, mesh = {Animals ; DNA, Mitochondrial/genetics ; Hemiptera/*microbiology ; Multilocus Sequence Typing ; Phylogeny ; Wolbachia/drug effects/genetics/*physiology ; }, abstract = {Wolbachia is the most widespread intracellular α-proteobacteria maternally inherited endosymbiont of insects and nematodes. These bacteria are associated with a number of different reproductive phenotypes of their hosts. Relatively few studies have dealt with distribution of infections across populations and with the influence of these bacteria on host genetic diversification and speciation. The aims of this study are to determine the distribution and rate of infection and to characterize the Wolbachia strains associated with Philaenus spumarius spittlebug (Hemiptera) by using multilocus sequencing typing (MLST) analysis and host phylogeography. The results showed that infection rate was significantly different between members of both main mitochondrial phylogenetic lineages of P. spumarius. We detected much higher infection rates of Wolbachia in P. spumarius populations from the north-east clade than the south-west clade. Moreover, the frequency of these infections varied within and outside the contact zone known from the Carpathians. Given the reproductive alterations which are often associated with this endosymbiont, Wolbachia probably maintain genetic differentiation of its hosts in its contact zone in the Carpathians. This is one of the first studies demonstrating the presence of Wolbachia across a large part of the range of insect species, including the contact zone. The spread of Wolbachia in P. spumarius populations can potentially cause speciation by compromising the potential reproductive barrier between infected and uninfected populations. We discuss possible implications of Wolbachia infection inducing cytoplasmic incompatibility in the population dynamics of this spittlebug but confirm that more studies are also required.}, } @article {pmid25675099, year = {2015}, author = {Ahmed, MZ and Li, SJ and Xue, X and Yin, XJ and Ren, SX and Jiggins, FM and Greeff, JM and Qiu, BL}, title = {The intracellular bacterium Wolbachia uses parasitoid wasps as phoretic vectors for efficient horizontal transmission.}, journal = {PLoS pathogens}, volume = {10}, number = {2}, pages = {e1004672}, pmid = {25675099}, issn = {1553-7374}, mesh = {Animals ; Cytoplasm/*metabolism ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Multilocus Sequence Typing/methods ; Wasps/genetics/*microbiology ; *Wolbachia/genetics ; }, abstract = {Facultative bacterial endosymbionts are associated with many arthropods and are primarily transmitted vertically from mother to offspring. However, phylogenetic affiliations suggest that horizontal transmission must also occur. Such horizontal transfer can have important biological and agricultural consequences when endosymbionts increase host fitness. So far horizontal transmission is considered rare and has been difficult to document. Here, we use fluorescence in situ hybridization (FISH) and multi locus sequence typing (MLST) to reveal a potentially common pathway of horizontal transmission of endosymbionts via parasitoids of insects. We illustrate that the mouthparts and ovipositors of an aphelinid parasitoid become contaminated with Wolbachia when this wasp feeds on or probes Wolbachia-infected Bemisia tabaci AsiaII7, and non-lethal probing of uninfected B. tabaci AsiaII7 nymphs by parasitoids carrying Wolbachia resulted in newly and stably infected B. tabaci matrilines. After they were exposed to infected whitefly, the parasitoids were able to transmit Wolbachia efficiently for the following 48 h. Whitefly infected with Wolbachia by parasitoids had increased survival and reduced development times. Overall, our study provides evidence for the horizontal transmission of Wolbachia between insect hosts by parasitic wasps, and the enhanced survival and reproductive abilities of insect hosts may adversely affect biological control programs.}, } @article {pmid25671520, year = {2015}, author = {Roy, V and Girondot, M and Harry, M}, title = {The distribution of Wolbachia in Cubitermes (Termitidae, Termitinae) castes and colonies: a modelling approach.}, journal = {PloS one}, volume = {10}, number = {2}, pages = {e0116070}, pmid = {25671520}, issn = {1932-6203}, mesh = {Algorithms ; Animals ; Female ; Isoptera/*microbiology ; Male ; *Models, Theoretical ; *Symbiosis ; Wolbachia/*classification ; }, abstract = {Wolbachia are endosymbiotic bacteria of arthropods and nematodes that are able to manipulate host reproduction. Although vertically transmitted via the cytoplasm in eggs, horizontal transmission of Wolbachia among and within arthropod species has been shown to be common. Eusocial insects represent interesting models for studying Wolbachia transmission due to colonial organization and close interaction between nestmates. Here we conducted a detailed screening of Wolbachia infection for 15 colonies of the very common soil-feeding termites Cubitermes spp. affinis subarquatus (Termitidae, Termitinae) that consist of four distinct phylogenetic species in the Lopé forest Reserve, Gabon. Infection tests showed that 50% of the individuals were Wolbachia positive (N = 555) with 90% of reproductives and 48% of offspring infected. White soldiers, which are transitional stages preceding mature soldiers, had a significantly higher mean infection rate (74%) than the other castes and stages (63%, 33% and 39% for larvae, workers and mature soldiers, respectively). We used a maximum likelihood method and Akaike's Information Criterion in order to explain the non-expected high rate of Wolbachia infection in white soldiers. The best model included a species effect for the stochastic loss of Wolbachia and a caste effect for the rate of gain. After fitting, the best model selected was for a species-specific rate of loss with a null rate of new gain for larvae, workers and soldiers and a probability of 0.72 whatever the species, that a white soldier becomes newly contaminated during that stage. The mean expected infection rate in white soldiers without a new gain was estimated to 17% instead of the 74% observed. Here we discuss the possible explanations to the high infection rate observed in white soldiers.}, } @article {pmid25668183, year = {2015}, author = {Gerardo, NM}, title = {Harnessing evolution to elucidate the consequences of symbiosis.}, journal = {PLoS biology}, volume = {13}, number = {2}, pages = {e1002066}, pmid = {25668183}, issn = {1545-7885}, mesh = {Animals ; Aphids/immunology/*microbiology ; Biological Evolution ; Buchnera/genetics/growth & development/*pathogenicity ; Drosophila melanogaster/immunology/*microbiology ; Enterobacteriaceae/genetics/growth & development/*pathogenicity ; Gene Dosage ; *Genome, Bacterial ; Genotype ; Longevity ; Phenotype ; Selection, Genetic ; Symbiosis/*genetics ; Virulence ; Wolbachia/genetics/growth & development/*pathogenicity ; }, abstract = {Many organisms harbor microbial associates that have profound impacts on host traits. The phenotypic effect of symbionts on their hosts may include changes in development, reproduction, longevity, and defense against natural enemies. Determining the consequences of associating with a microbial symbiont requires experimental comparison of hosts with and without symbionts. Then, determining the mechanism by which symbionts alter these phenotypes can involve genomic, genetic, and evolutionary approaches; however, many host-associated symbionts are not amenable to genetic approaches that require cultivation of the microbe outside the host. In the current issue of PLOS Biology, Chrostek and Teixeira highlight an elegant approach to studying functional mechanisms of symbiont-conferred traits. They used directed experimental evolution to select for strains of Wolbachia wMelPop (a bacterial symbiont of fruit flies) that differed in copy number of a region of the genome suspected to underlie virulence. Copy number evolved rapidly when under selection, and wMelPop strains with more copies of the region shortened the lives of their Drosophila hosts more than symbionts with fewer copies. Interestingly, the wMelPop strains with more copies also increase host resistance to viruses compared to symbionts with fewer copies. Their study highlights the power of exploiting alternative approaches when elucidating the functional impacts of symbiotic associations.}, } @article {pmid25668031, year = {2015}, author = {Chrostek, E and Teixeira, L}, title = {Mutualism breakdown by amplification of Wolbachia genes.}, journal = {PLoS biology}, volume = {13}, number = {2}, pages = {e1002065}, pmid = {25668031}, issn = {1545-7885}, support = {/WT_/Wellcome Trust/United Kingdom ; 094664/Z/10/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Biological Evolution ; Colony Count, Microbial ; Drosophila melanogaster/immunology/*microbiology ; Gene Dosage ; *Genome, Bacterial ; Genotype ; Longevity ; Multigene Family ; Phenotype ; Selection, Genetic ; Symbiosis/*genetics ; Virulence ; Wolbachia/*genetics/growth & development/*pathogenicity ; }, abstract = {Most insect species are associated with vertically transmitted endosymbionts. Because of the mode of transmission, the fitness of these symbionts is dependent on the fitness of the hosts. Therefore, these endosymbionts need to control their proliferation in order to minimize their cost for the host. The genetic bases and mechanisms of this regulation remain largely undetermined. The maternally inherited bacteria of the genus Wolbachia are the most common endosymbionts of insects, providing some of them with fitness benefits. In Drosophila melanogaster, Wolbachia wMelPop is a unique virulent variant that proliferates massively in the hosts and shortens their lifespan. The genetic bases of wMelPop virulence are unknown, and their identification would allow a better understanding of how Wolbachia levels are regulated. Here we show that amplification of a region containing eight Wolbachia genes, called Octomom, is responsible for wMelPop virulence. Using Drosophila lines selected for carrying Wolbachia with different Octomom copy numbers, we demonstrate that the number of Octomom copies determines Wolbachia titers and the strength of the lethal phenotype. Octomom amplification is unstable, and reversion of copy number to one reverts all the phenotypes. Our results provide a link between genotype and phenotype in Wolbachia and identify a genomic region regulating Wolbachia proliferation. We also prove that these bacteria can evolve rapidly. Rapid evolution by changes in gene copy number may be common in endosymbionts with a high number of mobile elements and other repeated regions. Understanding wMelPop pathogenicity and variability also allows researchers to better control and predict the outcome of releasing mosquitoes transinfected with this variant to block human vector-borne diseases. Our results show that transition from a mutualist to a pathogen may occur because of a single genomic change in the endosymbiont. This implies that there must be constant selection on endosymbionts to control their densities.}, } @article {pmid25658559, year = {2015}, author = {Chen, XP and Dong, YJ and Guo, WP and Wang, W and Li, MH and Xu, J and Dumler, JS and Zhang, YZ}, title = {Detection of Wolbachia genes in a patient with non-Hodgkin's lymphoma.}, journal = {Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases}, volume = {21}, number = {2}, pages = {182.e1-4}, doi = {10.1016/j.cmi.2014.09.008}, pmid = {25658559}, issn = {1469-0691}, mesh = {Bacteremia/*diagnosis/drug therapy ; Cluster Analysis ; DNA, Bacterial/*blood/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Fluoroquinolones/therapeutic use ; Gram-Negative Bacterial Infections/*diagnosis/drug therapy ; Humans ; Lymphoma, Non-Hodgkin/*complications ; Male ; Middle Aged ; Molecular Sequence Data ; Periplasmic Binding Proteins/genetics ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Treatment Outcome ; Wolbachia/*genetics ; }, abstract = {Wolbachia 16S rRNA and fbpA genes were twice detected over 5 days in the blood of a patient with high fever. The patient was given fluoroquinolones and the fever resolved. Four weeks later, he was diagnosed with non-Hodgkin's lymphoma and received R-CHOP (Rituximab, Cyclophosphamide, Doxorubicin, Vincristine, Prednisolone) treatment resulting in complete remission. This is the first report of detection of Wolbachia genes from the blood of human patients with non-Hodgkin's lymphoma.}, } @article {pmid25649218, year = {2015}, author = {Chagas-Moutinho, VA and Silva, R and de Souza, W and Motta, MC}, title = {Identification and ultrastructural characterization of the Wolbachia symbiont in Litomosoides chagasfilhoi.}, journal = {Parasites & vectors}, volume = {8}, number = {}, pages = {74}, pmid = {25649218}, issn = {1756-3305}, mesh = {Animals ; Female ; Filarioidea/*microbiology/physiology ; Male ; Microscopy, Electron, Transmission ; Phylogeny ; Subcutaneous Tissue/microbiology ; *Symbiosis ; Wolbachia/genetics/*isolation & purification/*physiology/ultrastructure ; }, abstract = {BACKGROUND: Filarial nematodes are arthropod-transmitted parasites of vertebrates that affect more than 150 million people around the world and remain a major public health problem throughout tropical and subtropical regions. Despite the importance of these nematodes, the current treatment strategies are not efficient in eliminating the parasite. The main strategy of control is based on chemotherapy with diethylcarbamazine, albendazole and ivermectin. In the 1970s, it was found that some filarids possess endosymbiotic bacteria that are important for the development, survival and infectivity of the nematodes. These bacteria belong to the genus Wolbachia, which is a widespread and abundant intracellular symbiont in worms. Knowledge about the structure of the bacteria and their relationship with their nematode hosts may allow new perspectives for the control of filarial nematodes.

METHODS: In this study, we used transmission electron microscopy combined with three-dimensional approaches to observe the structure of the endosymbiont of the filarial nematode Litomosoides chagasfilhoi, an experimental model for the study of lymphatic filariasis. In addition, the bacterium was classified based on PCR analyses.

RESULTS: The bacterium was mainly found in the hypodermis and in the female reproductive system in close association with host cell structures, such as the nucleus and endoplasmic reticulum. Our ultrastructural data also showed that the symbiont envelope is composed of two membrane units and is enclosed in a cytoplasmic vacuole, the symbiosome. Molecular data revealed that the bacterium of L. chagasfilhoi shares 100% identity with the Wolbachia endosymbiont of Litomosoides galizai.

CONCLUSIONS: Here we described ultrastructural aspects of the relationship of the Wolbachia with the filarial nematode Litomosoides chagasfilhoi and the findings lead us to consider this relationship as a mutualistic symbiosis.}, } @article {pmid25645184, year = {2015}, author = {Ndii, MZ and Hickson, RI and Allingham, D and Mercer, GN}, title = {Modelling the transmission dynamics of dengue in the presence of Wolbachia.}, journal = {Mathematical biosciences}, volume = {262}, number = {}, pages = {157-166}, doi = {10.1016/j.mbs.2014.12.011}, pmid = {25645184}, issn = {1879-3134}, mesh = {Aedes/*microbiology/*virology ; Animals ; Dengue/epidemiology/prevention & control/*transmission ; Humans ; Longevity ; Mathematical Concepts ; Models, Biological ; Pest Control, Biological ; Wolbachia/*physiology ; }, abstract = {Use of the bacterium Wolbachia is an innovative new strategy designed to break the cycle of dengue transmission. There are two main mechanisms by which Wolbachia could achieve this: by reducing the level of dengue virus in the mosquito and/or by shortening the host mosquito's lifespan. However, although Wolbachia shortens the lifespan, it also gives a breeding advantage which results in complex population dynamics. This study focuses on the development of a mathematical model to quantify the effect on human dengue cases of introducing Wolbachia into the mosquito population. The model consists of a compartment-based system of first-order differential equations; seasonal forcing in the mosquito population is introduced through the adult mosquito death rate. The analysis focuses on a single dengue outbreak typical of a region with a strong seasonally-varying mosquito population. We found that a significant reduction in human dengue cases can be obtained provided that Wolbachia-carrying mosquitoes persist when competing with mosquitoes without Wolbachia. Furthermore, using the Wolbachia strain WMel reduces the mosquito lifespan by at most 10% and allows them to persist in competition with non-Wolbachia-carrying mosquitoes. Mosquitoes carrying the WMelPop strain, however, are not likely to persist as it reduces the mosquito lifespan by up to 50%. When all other effects of Wolbachia on the mosquito physiology are ignored, cytoplasmic incompatibility alone results in a reduction in the number of human dengue cases. A sensitivity analysis of the parameters in the model shows that the transmission probability, the biting rate and the average adult mosquito death rate are the most important parameters for the outcome of the cumulative proportion of human individuals infected with dengue.}, } @article {pmid25622966, year = {2015}, author = {Sun, JT and Wang, MM and Zhang, YK and Chapuis, MP and Jiang, XY and Hu, G and Yang, XM and Ge, C and Xue, XF and Hong, XY}, title = {Evidence for high dispersal ability and mito-nuclear discordance in the small brown planthopper, Laodelphax striatellus.}, journal = {Scientific reports}, volume = {5}, number = {}, pages = {8045}, pmid = {25622966}, issn = {2045-2322}, mesh = {Animals ; Bayes Theorem ; DNA, Mitochondrial/chemistry/metabolism ; Electron Transport Complex IV/genetics ; Genetic Markers/*genetics ; Genetic Variation ; Genotype ; Hemiptera/*genetics/parasitology ; Logistic Models ; Metagenomics ; Microsatellite Repeats/genetics ; Mitochondria/*genetics ; Principal Component Analysis ; Sequence Analysis, DNA ; Wolbachia/genetics/physiology ; }, abstract = {Understanding dispersal ability in pest species is critical for both theoretical aspects of evolutionary and population biology and from a practical standpoint, such as implementing effective forecasting systems. The small brown planthopper (SBPH), Laodelphax striatellus (Fallén), is an economically important pest, but few data exist on its dispersal ability. Here, we used mitochondrial and nuclear markers to elucidate the population genetic structure of SBPH and of the parasitic bacterium Wolbachia throughout temperate and subtropical China. Our results showed that the SBPH populations in China lack significant differences in genetic structure, suggesting extensive gene flow. Multilocus sequence typing revealed that Wolbachia infection was systematic and due to the same strain (wStri) within and across populations. However, the mtDNA haplogroups had a nonrandom distribution across the sampling localities, which correlated to latitudinal and climatic gradients. We explain this mito-nuclear discordance as a result of historical population recolonization or mitochondria adaptation to climate.}, } @article {pmid25613091, year = {2015}, author = {White, JA and Richards, NK and Laugraud, A and Saeed, A and Curry, MM and McNeill, MR}, title = {Endosymbiotic candidates for parasitoid defense in exotic and native New Zealand weevils.}, journal = {Microbial ecology}, volume = {70}, number = {1}, pages = {274-286}, pmid = {25613091}, issn = {1432-184X}, mesh = {Animals ; Base Sequence ; DNA Primers/genetics ; Geography ; Likelihood Functions ; Metagenomics ; Models, Genetic ; Molecular Sequence Data ; New Zealand ; Phylogeny ; Polymerase Chain Reaction ; Rickettsia/*genetics ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Weevils/*microbiology ; Wolbachia/*genetics ; }, abstract = {Some insects are infected with maternally inherited bacterial endosymbionts that protect them against pathogens or parasitoids. The weevil Sitona obsoletus (=Sitona lepidus) is invasive in New Zealand, and suspected to contain such defensive symbionts, because it is particularly resistant to a Moroccan strain of the parasitoid Microctonus aethiopoides (which successfully attacks many other weevil species), and shows geographic variation in susceptibility to an Irish strain of the same parasitoid. Using 454 pyrosequencing, we investigated the bacterial community associated with S. obsoletus, two other exotic weevils (Sitona discoideus and Listronotus bonariensis) and two endemic New Zealand weevils (Irenimus aequalis and Steriphus variabilis). We found that S. obsoletus was infected by one strain of Wolbachia and two strains of Rickettsia, none of which were found in any other weevil species examined. Using diagnostic PCR, we found that S. obsoletus in the Northland region, where parasitism is highly variable, were primarily infected with Wolbachia and Rickettsia strain 2, indicating that these two symbionts should be investigated for potential defensive properties. In comparison, S. discoideus lacked any apparent maternally inherited bacterial endosymbionts. In the other weevil species, we found a different strain of Wolbachia and two different strains of Spiroplasma. Two weevil species (St. variabilis and L. bonariensis) were infected with distinct strains of Nardonella, the ancestral endosymbiont of weevils, whereas three weevil species (S. obsoletus, S. discoideus, and I. aequalis) lacked evidence for Nardonella infection. However, I. aequalis was consistently infected with a novel Enterobacteriaceae strain, suggesting that a symbiont replacement may have taken place, similar to that described for other weevil clades.}, } @article {pmid25611587, year = {2015}, author = {Gondorf, F and Berbudi, A and Buerfent, BC and Ajendra, J and Bloemker, D and Specht, S and Schmidt, D and Neumann, AL and Layland, LE and Hoerauf, A and Hübner, MP}, title = {Chronic filarial infection provides protection against bacterial sepsis by functionally reprogramming macrophages.}, journal = {PLoS pathogens}, volume = {11}, number = {1}, pages = {e1004616}, pmid = {25611587}, issn = {1553-7374}, mesh = {Animals ; Chronic Disease ; Coinfection ; Escherichia coli/*immunology ; Escherichia coli Infections/*immunology/prevention & control ; Female ; Filariasis/*immunology ; Filarioidea/*immunology/microbiology ; Gene Expression Regulation/immunology ; Macrophages, Peritoneal/*immunology/*metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Sepsis/immunology/*prevention & control ; Wolbachia/immunology ; }, abstract = {Helminths immunomodulate their hosts and induce a regulatory, anti-inflammatory milieu that prevents allergies and autoimmune diseases. Helminth immunomodulation may benefit sepsis outcome by preventing exacerbated inflammation and severe pathology, but the influence on bacterial clearance remains unclear. To address this, mice were chronically infected with the filarial nematode Litomosoides sigmodontis (L.s.) and the outcome of acute systemic inflammation caused by i.p. Escherichia coli injection was determined. L.s. infection significantly improved E. coli-induced hypothermia, bacterial clearance and sepsis survival and correlated with reduced concentrations of associated pro-inflammatory cytokines/chemokines and a less pronounced pro-inflammatory macrophage gene expression profile. Improved sepsis outcome in L.s.-infected animals was mediated by macrophages, but independent of the alternatively activated macrophage subset. Endosymbiotic Wolbachia bacteria that are present in most human pathogenic filariae, as well as L.s., signal via TLR2 and modulate macrophage function. Here, gene expression profiles of peritoneal macrophages from L.s.-infected mice revealed a downregulation of genes involved in TLR signaling, and pulsing of macrophages in vitro with L.s. extract reduced LPS-triggered activation. Subsequent transfer improved sepsis outcome in naïve mice in a Wolbachia- and TLR2-dependent manner. In vivo, phagocytosis was increased in macrophages from L.s.-infected wild type, but not TLR2-deficient animals. In association, L.s. infection neither improved bacterial clearance in TLR2-deficient animals nor ameliorated E. coli-induced hypothermia and sepsis survival. These results indicate that chronic L.s. infection has a dual beneficial effect on bacterial sepsis, reducing pro-inflammatory immune responses and improving bacterial control. Thus, helminths and their antigens may not only improve the outcome of autoimmune and allergic diseases, but may also present new therapeutic approaches for acute inflammatory diseases that do not impair bacterial control.}, } @article {pmid25596936, year = {2015}, author = {Shankar, P and Kulkarni, VM and Kumar, LS}, title = {Male biased gene flow in banana pseudostem weevil (Odoiporus longicollis Oliver) as revealed by analysis of the COI-tRNA(Leu) COII region.}, journal = {Genetica}, volume = {143}, number = {1}, pages = {85-92}, pmid = {25596936}, issn = {1573-6857}, mesh = {Animals ; Base Composition ; *DNA, Mitochondrial ; *Gene Flow ; Genetic Variation ; Geography ; Haplotypes ; India ; Male ; Phylogeny ; RNA, Transfer, Leu/*genetics ; Sequence Analysis, DNA ; Weevils/classification/*genetics ; }, abstract = {The genetic diversity amongst thirty weevils representing six Indian populations of banana pseudostem weevil, i.e., Odoiporus longicollis (Oliver) was estimated by sequence analysis of the partial COI-tRNA(Leu)-COII region. The sequences exhibited AT bias typical of insect mitochondrial DNA which was highest in the first codon position of COI and in the third codon position of COII. There was no phylogeographic distribution of the populations. The Fu and Li's D and F tests were non-significant for this mitochondrial region. No Wolbachia infection was detected in any of the populations. The genetic differentiation amongst the populations was highly significant (p < 0.001; χ2 = 123.333; df = 75), suggesting restricted gene flow between the populations. This result did not correlate with that obtained with nuclear rDNA markers, i.e., ITS1 and ITS2, suggesting a male biased gene flow between the populations.}, } @article {pmid25593788, year = {2014}, author = {Gordon, JE and Christie, PJ}, title = {The Agrobacterium Ti Plasmids.}, journal = {Microbiology spectrum}, volume = {2}, number = {6}, pages = {}, pmid = {25593788}, issn = {2165-0497}, support = {R01 GM048746/GM/NIGMS NIH HHS/United States ; GM48746/GM/NIGMS NIH HHS/United States ; }, mesh = {Agrobacterium tumefaciens/*genetics ; DNA Replication ; DNA, Bacterial/genetics/metabolism ; Plant Diseases/microbiology ; *Plant Tumor-Inducing Plasmids ; Plants/microbiology ; }, abstract = {Agrobacterium tumefaciens is a plant pathogen with the capacity to deliver a segment of oncogenic DNA carried on a large plasmid called the tumor-inducing or Ti plasmid to susceptible plant cells. A. tumefaciens belongs to the class Alphaproteobacteria, whose members include other plant pathogens (Agrobacterium rhizogenes), plant and insect symbionts (Rhizobium spp. and Wolbachia spp., respectively), human pathogens (Brucella spp., Bartonella spp., Rickettsia spp.), and nonpathogens (Caulobacter crescentus, Rhodobacter sphaeroides). Many species of Alphaproteobacteria carry large plasmids ranging in size from ∼100 kb to nearly 2 Mb. These large replicons typically code for functions essential for cell physiology, pathogenesis, or symbiosis. Most of these elements rely on a conserved gene cassette termed repABC for replication and partitioning, and maintenance at only one or a few copies per cell. The subject of this review is the ∼200-kb Ti plasmids carried by infectious strains of A. tumefaciens. We will summarize the features of this plasmid as a representative of the repABC family of megaplasmids. We will also describe novel features of this plasmid that enable A. tumefaciens cells to incite tumor formation in plants, sense and respond to an array of plant host and bacterial signal molecules, and maintain and disseminate the plasmid among populations of agrobacteria. At the end of this review, we will describe how this natural genetic engineer has been adapted to spawn an entire industry of plant biotechnology and review its potential for use in future therapeutic applications of plant and nonplant species.}, } @article {pmid25589901, year = {2015}, author = {Borkowski, PK and Rymkiewicz, G and Golebiewska, J and Nestoros, N and Romejko-Jarosinska, J and Zarnowska-Prymek, H and Masny, A and Palucki, J and Cielecka, D}, title = {The first case of human autochtonous subconjunctival dirofilariosis in Poland and MALT lymphoma as possible consequence of this parasitosis.}, journal = {Infectious agents and cancer}, volume = {10}, number = {1}, pages = {1}, pmid = {25589901}, issn = {1750-9378}, abstract = {The first case of human dirofilarosis in Poland was recorded in 2007. Until that time our country was free of Dirofilaria repens. Recent studies show that 21,4- 60% of dogs in Warsaw region harbour microfilariae, therefore it is becoming a growing problem in Central Europe. In April 2013 a subconjunctival D. repens was removed from the eye of 61-year-old woman. It was the twenty first case of this disease in Poland, the third case of eye dirofilaria and the fourth autochtonous case. The patient had never been abroad, so it was the first case of autochtonous human ocular dirofilariosis in Poland. Nine months after the D. repens had been removed, a MALT lymphoma was discovered. In the article we discuss whether a MALT lymphoma of the lacrimal gland of the eye, previously affected by the parasite, may be the consequence of the invasion.}, } @article {pmid25579393, year = {2015}, author = {Voronin, D and Abeykoon, AM and Gunawardene, YI and Dassanayake, RS}, title = {Absence of Wolbachia endobacteria in Sri Lankan isolates of the nematode parasite of animals Setaria digitata.}, journal = {Veterinary parasitology}, volume = {207}, number = {3-4}, pages = {350-354}, doi = {10.1016/j.vetpar.2014.12.024}, pmid = {25579393}, issn = {1873-2550}, mesh = {Animals ; Cattle/parasitology ; Genes, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Setaria Nematode/*microbiology ; Sri Lanka ; Wolbachia/genetics/*physiology ; }, abstract = {Setaria digitata is an animal filarial parasite with natural hosts of cattle and buffaloes that causes mild disease conditions. Infection of non-permissive hosts such as goats, sheep and horses, by this nematode can cause cerebrospinal nematodiasis that leads to lumbar paralysis and the eventual death of the animals and inflicts considerable economic losses on livestock farmers. Wolbachia are obligate mutualistic endosymbionts for some filarial nematodes and are currently being targeted for the control of diseases caused by these parasites. However, little is known about the occurrence of this endosymbiont in the Setariidae family. In this work, worms collected from infected cattle in Sri Lanka were morphologically identified as S. digitata and tested for the presence of Wolbachia by PCR screening using the WSP- and Wolbachia-specific 16S rRNA and multilocus sequence typing primers that were designed to amplify the gatB, coxA, hcpA, ftsZ and fbpA sequences of Wolbachia. The presence of endobacteria in S. digitata was also examined by whole-mount immunofluorescence staining of the parasites and transmission electron microscopic studies. These analyses did not produce evidence of presence of Wolbachia or any other endosymbiotic bacteria in S. digitata, whereas such evidence was found in Brugia malayi, which was used as a positive control in this study.}, } @article {pmid25575310, year = {2015}, author = {Cohen, C and Toh, E and Munro, D and Dong, Q and Hawlena, H}, title = {Similarities and seasonal variations in bacterial communities from the blood of rodents and from their flea vectors.}, journal = {The ISME journal}, volume = {9}, number = {7}, pages = {1662-1676}, pmid = {25575310}, issn = {1751-7370}, mesh = {Animals ; Bacterial Infections/microbiology/*veterinary ; Bartonella/isolation & purification ; Female ; Flea Infestations/*veterinary ; Gerbillinae/*blood/microbiology ; Insect Vectors ; Male ; *Seasons ; Siphonaptera/*microbiology ; Wolbachia/isolation & purification ; }, abstract = {Vector-borne microbes are subject to the ecological constraints of two distinct microenvironments: that in the arthropod vector and that in the blood of its vertebrate host. Because the structure of bacterial communities in these two microenvironments may substantially affect the abundance of vector-borne microbes, it is important to understand the relationship between bacterial communities in both microenvironments and the determinants that shape them. We used pyrosequencing analyses to compare the structure of bacterial communities in Synosternus cleopatrae fleas and in the blood of their Gerbillus andersoni hosts. We also monitored the interindividual and seasonal variability in these bacterial communities by sampling the same individual wild rodents during the spring and again during the summer. We show that the bacterial communities in each sample type (blood, female flea or male flea) had a similar phylotype composition among host individuals, but exhibited seasonal variability that was not directly associated with host characteristics. The structure of bacterial communities in male fleas and in the blood of their rodent hosts was remarkably similar and was dominated by flea-borne Bartonella and Mycoplasma phylotypes. A lower abundance of flea-borne bacteria and the presence of Wolbachia phylotypes distinguished bacterial communities in female fleas from those in male fleas and in rodent blood. These results suggest that the overall abundance of a certain vector-borne microbe is more likely to be determined by the abundance of endosymbiotic bacteria in the vector, abundance of other vector-borne microbes co-occurring in the vector and in the host blood and by seasonal changes, than by host characteristics.}, } @article {pmid25569144, year = {2015}, author = {Westblade, LF and Fischer, PU and Haghi, N and Schniederjan, MJ and Pritt, BS and Long, JG and Jerris, RC and Garola, RE}, title = {A five-year-old child with a subcutaneous forehead nodule.}, journal = {Pediatric and developmental pathology : the official journal of the Society for Pediatric Pathology and the Paediatric Pathology Society}, volume = {18}, number = {2}, pages = {164-166}, doi = {10.2350/14-08-1545-CR.1}, pmid = {25569144}, issn = {1093-5266}, mesh = {Animals ; Antiparasitic Agents/therapeutic use ; Biopsy ; Child, Preschool ; Female ; Forehead ; Humans ; Immunohistochemistry ; Ivermectin/therapeutic use ; Onchocerca/*isolation & purification ; Onchocerciasis/diagnosis/*parasitology/therapy ; Predictive Value of Tests ; Subcutaneous Tissue/*parasitology/pathology ; Treatment Outcome ; }, abstract = {We describe a case of a 5-year-old girl with onchocerciasis. The patient was recently adopted from Ethiopia and presented with a firm, raised nodule on the midportion of the forehead. Initially, Langerhans cell histiocytosis with bone involvement was suspected; however, histopathologic analysis of the excised nodule revealed the presence of a young-adult, female Onchocerca volvulus worm. This case exemplifies the importance of recognizing the key morphologic characteristics of adult O. volvulus worms isolated from pediatric patients in nonendemic areas to ensure adroit clinical management.}, } @article {pmid25541991, year = {2014}, author = {Huemer, P and Mutanen, M and Sefc, KM and Hebert, PD}, title = {Testing DNA barcode performance in 1000 species of European lepidoptera: large geographic distances have small genetic impacts.}, journal = {PloS one}, volume = {9}, number = {12}, pages = {e115774}, pmid = {25541991}, issn = {1932-6203}, mesh = {Animals ; Biodiversity ; Cluster Analysis ; DNA Barcoding, Taxonomic/*methods ; *Geography ; Lepidoptera/*classification ; }, abstract = {This study examines the performance of DNA barcodes (mt cytochrome c oxidase 1 gene) in the identification of 1004 species of Lepidoptera shared by two localities (Finland, Austria) that are 1600 km apart. Maximum intraspecific distances for the pooled data were less than 2% for 880 species (87.6%), while deeper divergence was detected in 124 species. Despite such variation, the overall DNA barcode library possessed diagnostic COI sequences for 98.8% of the taxa. Because a reference library based on Finnish specimens was highly effective in identifying specimens from Austria, we conclude that barcode libraries based on regional sampling can often be effective for a much larger area. Moreover, dispersal ability (poor, good) and distribution patterns (disjunct, fragmented, continuous, migratory) had little impact on levels of intraspecific geographic divergence. Furthermore, the present study revealed that, despite the intensity of past taxonomic work on European Lepidoptera, nearly 20% of the species shared by Austria and Finland require further work to clarify their status. Particularly discordant BIN (Barcode Index Number) cases should be checked to ascertain possible explanatory factors such as incorrect taxonomy, hybridization, introgression, and Wolbachia infections.}, } @article {pmid25541519, year = {2015}, author = {Glowska, E and Dragun-Damian, A and Dabert, M and Gerth, M}, title = {New Wolbachia supergroups detected in quill mites (Acari: Syringophilidae).}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {30}, number = {}, pages = {140-146}, doi = {10.1016/j.meegid.2014.12.019}, pmid = {25541519}, issn = {1567-7257}, mesh = {Animals ; Birds/*parasitology ; DNA, Bacterial/analysis/genetics ; Feathers/*parasitology ; Female ; Male ; Mites/*microbiology ; Phylogeny ; Symbiosis ; *Wolbachia/classification/genetics/isolation & purification ; }, abstract = {Wolbachia is the most abundant intracellular bacterial genus infecting a wide range of arthropods and filarial nematodes. Wolbachia have evolved parasitic, mutualistic and commensal relationships with their hosts but in arthropods generally act as reproductive parasites, inducing a wide range of phenotypic effects such as cytoplasmic incompatibility, parthenogenesis, feminization and male-killing. Up to now, the genus has been divided into 14 supergroups successively named A-O. Here, we describe two new Wolbachia supergroups from syringophilid mites (Acari: Cheyletoidea). These obligatory ectoparasites of birds inhabit the quills of feathers in many avian groups. The species of this family reproduce in a haplodiploid mode sensu arrhenotoky and are usually strongly female-biased. Based on the sequences of four protein-coding genes (ftsZ, gltA and groEL and coxA) and the 16S rRNA we identified strains of three Wolbachia supergroups (F and two distinct, yet undescribed ones) in five quill mite species. Our results suggest that in some cases the distribution of the bacteria can be better correlated with the mite's bird host rather than with mite taxonomy as such. The discovery of two new Wolbachia supergroups not only broadens the knowledge of the diversity of this bacterium but also raises questions about potential effects induced in quill mites and transmission mechanisms of the endosymbionts in this peculiar bacteria-quill mite-bird system.}, } @article {pmid25540454, year = {2015}, author = {Baudet, C and Donati, B and Sinaimeri, B and Crescenzi, P and Gautier, C and Matias, C and Sagot, MF}, title = {Cophylogeny reconstruction via an approximate Bayesian computation.}, journal = {Systematic biology}, volume = {64}, number = {3}, pages = {416-431}, pmid = {25540454}, issn = {1076-836X}, mesh = {*Algorithms ; Animals ; Arthropods/classification/microbiology ; Bayes Theorem ; Classification/*methods ; Host-Parasite Interactions ; *Phylogeny ; Wolbachia/classification/physiology ; }, abstract = {Despite an increasingly vast literature on cophylogenetic reconstructions for studying host-parasite associations, understanding the common evolutionary history of such systems remains a problem that is far from being solved. Most algorithms for host-parasite reconciliation use an event-based model, where the events include in general (a subset of) cospeciation, duplication, loss, and host switch. All known parsimonious event-based methods then assign a cost to each type of event in order to find a reconstruction of minimum cost. The main problem with this approach is that the cost of the events strongly influences the reconciliation obtained. Some earlier approaches attempt to avoid this problem by finding a Pareto set of solutions and hence by considering event costs under some minimization constraints. To deal with this problem, we developed an algorithm, called Coala, for estimating the frequency of the events based on an approximate Bayesian computation approach. The benefits of this method are 2-fold: (i) it provides more confidence in the set of costs to be used in a reconciliation, and (ii) it allows estimation of the frequency of the events in cases where the data set consists of trees with a large number of taxa. We evaluate our method on simulated and on biological data sets. We show that in both cases, for the same pair of host and parasite trees, different sets of frequencies for the events lead to equally probable solutions. Moreover, often these solutions differ greatly in terms of the number of inferred events. It appears crucial to take this into account before attempting any further biological interpretation of such reconciliations. More generally, we also show that the set of frequencies can vary widely depending on the input host and parasite trees. Indiscriminately applying a standard vector of costs may thus not be a good strategy.}, } @article {pmid25538866, year = {2014}, author = {LePage, DP and Jernigan, KK and Bordenstein, SR}, title = {The relative importance of DNA methylation and Dnmt2-mediated epigenetic regulation on Wolbachia densities and cytoplasmic incompatibility.}, journal = {PeerJ}, volume = {2}, number = {}, pages = {e678}, pmid = {25538866}, issn = {2167-8359}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; }, abstract = {Wolbachia pipientis is a worldwide bacterial parasite of arthropods that infects germline cells and manipulates host reproduction to increase the ratio of infected females, the transmitting sex of the bacteria. The most common reproductive manipulation, cytoplasmic incompatibility (CI), is expressed as embryonic death in crosses between infected males and uninfected females. Specifically, Wolbachia modify developing sperm in the testes by unknown means to cause a post-fertilization disruption of the sperm chromatin that incapacitates the first mitosis of the embryo. As these Wolbachia-induced changes are stable, reversible, and affect the host cell cycle machinery including DNA replication and chromosome segregation, we hypothesized that the host methylation pathway is targeted for modulation during cytoplasmic incompatibility because it accounts for all of these traits. Here we show that infection of the testes is associated with a 55% increase of host DNA methylation in Drosophila melanogaster, but methylation of the paternal genome does not correlate with penetrance of CI. Overexpression and knock out of the Drosophila DNA methyltransferase Dnmt2 neither induces nor increases CI. Instead, overexpression decreases Wolbachia titers in host testes by approximately 17%, leading to a similar reduction in CI levels. Finally, strength of CI induced by several different strains of Wolbachia does not correlate with levels of DNA methylation in the host testes. We conclude that DNA methylation mediated by Drosophila's only known methyltransferase is not required for the transgenerational sperm modification that causes CI.}, } @article {pmid25537873, year = {2015}, author = {Walker, M and Specht, S and Churcher, TS and Hoerauf, A and Taylor, MJ and Basáñez, MG}, title = {Therapeutic efficacy and macrofilaricidal activity of doxycycline for the treatment of river blindness.}, journal = {Clinical infectious diseases : an official publication of the Infectious Diseases Society of America}, volume = {60}, number = {8}, pages = {1199-1207}, pmid = {25537873}, issn = {1537-6591}, support = {MR/K010174/1/MRC_/Medical Research Council/United Kingdom ; 092677/Z/10/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Anthelmintics/*pharmacology/*therapeutic use ; Clinical Trials as Topic ; Doxycycline/*pharmacology/*therapeutic use ; Female ; Humans ; Models, Statistical ; Onchocerca volvulus/*microbiology ; Onchocerciasis, Ocular/*drug therapy ; Treatment Outcome ; Wolbachia/*drug effects ; }, abstract = {BACKGROUND: Onchocerca volvulus and lymphatic filariae, causing river blindness and elephantiasis, depend on endosymbiotic Wolbachia bacteria for growth, development, fertility, and survival. Clinical trials have shown that doxycycline treatment eliminates Wolbachia, causing long-term sterilization of adult female filariae and effecting potent macrofilaricidal activity. The continual reinfection by drug-naive worms that occurs in these trial settings dilutes observable anti-Wolbachia and antifilarial effects, making it difficult to estimate therapeutic efficacy and compare different doxycycline regimens, evaluated at different times after treatment.

METHODS: A meta-analytical modeling framework is developed to link all usable data collected from clinical trials measuring the Wolbachia status and viability of individual female adult worms collected at various times after treatment with 4, 5, or 6 weeks of daily 100 or 200 mg oral doxycycline. The framework is used to estimate efficacy parameters that are not directly measurable as trial outcomes.

RESULTS: The estimated efficacy of doxycycline (the maximum proportional reduction in the percentage of adult female O. volvulus positive for Wolbachia) is 91%-94% on average, irrespective of the treatment regimen. Efficacy is >95% in the majority of trial participants. The life span of Wolbachia-depleted worms is reduced by 70%-80%, from approximately 10 years to 2-3 years.

CONCLUSIONS: The efficacy parameters are pertinent to the prospects of using doxycycline on a "test and treat" basis for onchocerciasis control and confirm doxycycline as a potent macrofilaricidal therapy. The modeling approach is more generally relevant to the design and evaluation of clinical trials for antifilarial drugs conducted in endemic settings.}, } @article {pmid25521705, year = {2014}, author = {Drinkwater, B and Charleston, MA}, title = {Introducing TreeCollapse: a novel greedy algorithm to solve the cophylogeny reconstruction problem.}, journal = {BMC bioinformatics}, volume = {15 Suppl 16}, number = {Suppl 16}, pages = {S14}, pmid = {25521705}, issn = {1471-2105}, mesh = {*Algorithms ; *Biological Evolution ; Computational Biology/*methods ; Humans ; Models, Theoretical ; *Phylogeny ; }, abstract = {BACKGROUND: Cophylogeny mapping is used to uncover deep coevolutionary associations between two or more phylogenetic histories at a macro coevolutionary scale. As cophylogeny mapping is NP-Hard, this technique relies heavily on heuristics to solve all but the most trivial cases. One notable approach utilises a metaheuristic to search only a subset of the exponential number of fixed node orderings possible for the phylogenetic histories in question. This is of particular interest as it is the only known heuristic that guarantees biologically feasible solutions. This has enabled research to focus on larger coevolutionary systems, such as coevolutionary associations between figs and their pollinator wasps, including over 200 taxa. Although able to converge on solutions for problem instances of this size, a reduction from the current cubic running time is required to handle larger systems, such as Wolbachia and their insect hosts.

RESULTS: Rather than solving this underlying problem optimally this work presents a greedy algorithm called TreeCollapse, which uses common topological patterns to recover an approximation of the coevolutionary history where the internal node ordering is fixed. This approach offers a significant speed-up compared to previous methods, running in linear time. This algorithm has been applied to over 100 well-known coevolutionary systems converging on Pareto optimal solutions in over 68% of test cases, even where in some cases the Pareto optimal solution has not previously been recoverable. Further, while TreeCollapse applies a local search technique, it can guarantee solutions are biologically feasible, making this the fastest method that can provide such a guarantee.

CONCLUSION: As a result, we argue that the newly proposed algorithm is a valuable addition to the field of coevolutionary research. Not only does it offer a significantly faster method to estimate the cost of cophylogeny mappings but by using this approach, in conjunction with existing heuristics, it can assist in recovering a larger subset of the Pareto front than has previously been possible.}, } @article {pmid25521619, year = {2014}, author = {Toomey, ME and Frydman, HM}, title = {Extreme divergence of Wolbachia tropism for the stem-cell-niche in the Drosophila testis.}, journal = {PLoS pathogens}, volume = {10}, number = {12}, pages = {e1004577}, pmid = {25521619}, issn = {1553-7374}, support = {1R56AI097589/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*microbiology ; Female ; Male ; Ovary/microbiology ; Phenotype ; Phylogeny ; Sex Factors ; Stem Cell Niche/physiology ; Testis/microbiology ; Tropism/physiology ; Wolbachia/*physiology ; }, abstract = {Microbial tropism, the infection of specific cells and tissues by a microorganism, is a fundamental aspect of host-microbe interactions. The intracellular bacteria Wolbachia have a peculiar tropism for the stem cell niches in the Drosophila ovary, the microenvironments that support the cells producing the eggs. The molecular underpinnings of Wolbachia stem cell niche tropism are unknown. We have previously shown that the patterns of tropism in the ovary show a high degree of conservation across the Wolbachia lineage, with closely related Wolbachia strains usually displaying the same pattern of stem cell niche tropism. It has also been shown that tropism to these structures in the ovary facilitates both vertical and horizontal transmission, providing a strong selective pressure towards evolutionary conservation of tropism. Here we show great disparity in the evolutionary conservation and underlying mechanisms of stem cell niche tropism between male and female gonads. In contrast to females, niche tropism in the male testis is not pervasive, present in only 45% of niches analyzed. The patterns of niche tropism in the testis are not evolutionarily maintained across the Wolbachia lineage, unlike what was shown in the females. Furthermore, hub tropism does not correlate with cytoplasmic incompatibility, a Wolbachia-driven phenotype imprinted during spermatogenesis. Towards identifying the molecular mechanism of hub tropism, we performed hybrid analyses of Wolbachia strains in non-native hosts. These results indicate that both Wolbachia and host derived factors play a role in the targeting of the stem cell niche in the testis. Surprisingly, even closely related Wolbachia strains in Drosophila melanogaster, derived from a single ancestor only 8,000 years ago, have significantly different tropisms to the hub, highlighting that stem cell niche tropism is rapidly diverging in males. These findings provide a powerful system to investigate the mechanisms and evolution of microbial tissue tropism.}, } @article {pmid25520356, year = {2015}, author = {Ponton, F and Wilson, K and Holmes, A and Raubenheimer, D and Robinson, KL and Simpson, SJ}, title = {Macronutrients mediate the functional relationship between Drosophila and Wolbachia.}, journal = {Proceedings. Biological sciences}, volume = {282}, number = {1800}, pages = {20142029}, pmid = {25520356}, issn = {1471-2954}, support = {BB/F004311/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Drosophila melanogaster/microbiology/*physiology ; Female ; Fertility/physiology ; Longevity/physiology ; Male ; Nutritional Physiological Phenomena ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited bacterial endosymbionts that naturally infect a diverse array of arthropods. They are primarily known for their manipulation of host reproductive biology, and recently, infections with Wolbachia have been proposed as a new strategy for controlling insect vectors and subsequent human-transmissible diseases. Yet, Wolbachia abundance has been shown to vary greatly between individuals and the magnitude of the effects of infection on host life-history traits and protection against infection is correlated to within-host Wolbachia abundance. It is therefore essential to better understand the factors that modulate Wolbachia abundance and effects on host fitness. Nutrition is known to be one of the most important mediators of host-symbiont interactions. Here, we used nutritional geometry to quantify the role of macronutrients on insect-Wolbachia relationships in Drosophila melanogaster. Our results show fundamental interactions between diet composition, host diet selection, Wolbachia abundance and effects on host lifespan and fecundity. The results and methods described here open a new avenue in the study of insect-Wolbachia relationships and are of general interest to numerous research disciplines, ranging from nutrition and life-history theory to public health.}, } @article {pmid25517525, year = {2014}, author = {Soares, HS and Camargo, LM and Gennari, SM and Labruna, MB}, title = {Survey of canine tick-borne diseases in Lábrea, Brazilian Amazon: 'accidental' findings of Dirofilaria immitis infection.}, journal = {Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria}, volume = {23}, number = {4}, pages = {473-480}, doi = {10.1590/S1984-29612014093}, pmid = {25517525}, issn = {1984-2961}, mesh = {Animals ; Brazil/epidemiology ; Dirofilariasis/*epidemiology ; Dog Diseases/*epidemiology/*parasitology ; Dogs ; Female ; Incidental Findings ; Male ; Tick-Borne Diseases/epidemiology/*veterinary ; }, abstract = {Blood samples were collected from 99 domestic dogs from the urban and rural areas of the Lábrea municipality, state of Amazonas, Brazil. Canine serum samples were tested by immunofluorescence assay against Rickettsia spp., which revealed that only 3.0% (1/33) and 7.6% (5/66) of the dogs from urban and rural areas, respectively, reacted positively to at least one Rickettsia species. DNA was extracted from canine blood and tested by a battery of PCR assays targeting protozoa of the genera Babesia and Hepatozoon, and bacteria of the genera Rickettsia and Ehrlichia and family Anaplasmataceae. All samples were negative in the PCR assays targeting the genera Babesia, Hepatozoon, Ehrlichia and Rickettsia. For Anaplasmataceae, 3% (1/33) and 39.4% (26/66) of the urban and rural dogs, respectively, yielded amplicons that generated DNA sequences 100% identical to the corresponding sequence of Wolbachia endosymbiont of Dirofilaria immitis. Because of these results, all canine DNA samples were further tested in a PCR assay targeting filarial nematodes, which was positive for 18.2% (6/33) and 57.6% (38/66) urban and rural dogs, respectively. Filarial-PCR products generated DNA sequences 100% identical to D. immitis. While tick-borne infections were rare in Lábrea, D. immitis infection rates were among the highest reported in South America.}, } @article {pmid25516838, year = {2014}, author = {Johnston, KL and Ford, L and Umareddy, I and Townson, S and Specht, S and Pfarr, K and Hoerauf, A and Altmeyer, R and Taylor, MJ}, title = {Repurposing of approved drugs from the human pharmacopoeia to target Wolbachia endosymbionts of onchocerciasis and lymphatic filariasis.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {4}, number = {3}, pages = {278-286}, pmid = {25516838}, issn = {2211-3207}, abstract = {Lymphatic filariasis and onchocerciasis are debilitating diseases caused by parasitic filarial nematodes infecting around 150 million people throughout the tropics with more than 1.5 billion at risk. As with other neglected tropical diseases, classical drug-discovery and development is lacking and a 50 year programme of macrofilaricidal discovery failed to deliver a drug which can be used as a public health tool. Recently, antibiotic targeting of filarial Wolbachia, an essential bacterial symbiont, has provided a novel drug treatment for filariasis with macrofilaricidal activity, although the current gold-standard, doxycycline, is unsuitable for use in mass drug administration (MDA). The anti-Wolbachia (A·WOL) Consortium aims to identify novel anti-Wolbachia drugs, compounds or combinations that are suitable for use in MDA. Development of a Wolbachia cell-based assay has enabled the screening of the approved human drug-pharmacopoeia (∼2600 drugs) for a potential repurposing. This screening strategy has revealed that approved drugs from various classes show significant bacterial load reduction equal to or superior to the gold-standard doxycycline, with 69 orally available hits from different drug categories being identified. Based on our defined hit criteria, 15 compounds were then selectively screened in a Litomosoides sigmodontis mouse model, 4 of which were active. These came from the tetracycline, fluoroquinolone and rifamycin classes. This strategy of repurposing approved drugs is a promising development in the goal of finding a novel treatment against filariasis and could also be a strategy applicable for other neglected tropical diseases.}, } @article {pmid25516837, year = {2014}, author = {Lustigman, S and Melnikow, E and Anand, SB and Contreras, A and Nandi, V and Liu, J and Bell, A and Unnasch, TR and Rogers, MB and Ghedin, E}, title = {Potential involvement of Brugia malayi cysteine proteases in the maintenance of the endosymbiotic relationship with Wolbachia.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {4}, number = {3}, pages = {267-277}, pmid = {25516837}, issn = {2211-3207}, support = {R01 AI072465/AI/NIAID NIH HHS/United States ; R56 AI101372/AI/NIAID NIH HHS/United States ; }, abstract = {Brugia malayi, a parasitic nematode that causes lymphatic filariasis, harbors endosymbiotic intracellular bacteria, Wolbachia, that are required for the development and reproduction of the worm. The essential nature of this endosymbiosis led to the development of anti-Wolbachia chemotherapeutic approaches for the treatment of human filarial infections. Our study is aimed at identifying specific proteins that play a critical role in this endosymbiotic relationship leading to the identification of potential targets in the adult worms. Filarial cysteine proteases are known to be involved in molting and embryogenesis, processes shown to also be Wolbachia dependent. Based on the observation that cysteine protease transcripts are differentially regulated in response to tetracycline treatment, we focused on defining their role in symbiosis. We observe a bimodal regulation pattern of transcripts encoding cysteine proteases when in vitro tetracycline treated worms were examined. Using tetracycline-treated infertile female worms and purified embryos we established that the first peak of the bimodal pattern corresponds to embryonic transcripts while the second takes place within the hypodermis of the adult worms. Localization studies of the native proteins corresponding to Bm-cpl-3 and Bm-cpl-6 indicate that they are present in the area surrounding Wolbachia, and, in some cases, the proteins appear localized within the bacteria. Both proteins were also found in the inner bodies of microfilariae. The possible role of these cysteine proteases during development and endosymbiosis was further characterized using RNAi. Reduction in Bm-cpl-3 and Bm-cpl-6 transcript levels was accompanied by hindered microfilarial development and release, and reduced Wolbachia DNA levels, making these enzymes strong drug target candidates.}, } @article {pmid25515563, year = {2014}, author = {Rong, X and Zhang, YK and Zhang, KJ and Hong, XY}, title = {Identification of Wolbachia-responsive microRNAs in the two-spotted spider mite, Tetranychus urticae.}, journal = {BMC genomics}, volume = {15}, number = {1}, pages = {1122}, pmid = {25515563}, issn = {1471-2164}, mesh = {Animals ; Apoptosis/genetics ; Carrier Proteins/genetics ; Female ; Gene Expression Regulation ; Gene Library ; Host-Pathogen Interactions/genetics ; Lysosomes/genetics/metabolism ; Male ; MicroRNAs/classification/genetics/*metabolism ; Reproduction/genetics ; Sphingolipids/genetics/metabolism ; Tetranychidae/*genetics ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: The two-spotted spider mite, Tetranychus urticae, is infected with Wolbachia, which have the ability to manipulate host reproduction and fitness. MicroRNAs (miRNAs) are small non-coding RNAs that are involved in many biological processes such as development, reproduction and host-pathogen interactions. Although miRNA was observed to involve in Wolbachia-host interactions in the other insect systems, its roles have not been fully deciphered in the two-spotted spider mite.

RESULTS: Small RNA libraries of infected and uninfected T. urticae for both sexes (in total four libraries) were constructed. By integrating the mRNA data originated from the same samples, the target genes of the differentially expressed miRNAs were predicted. Then, GO and pathway analyses were performed for the target genes. Comparison of libraries showed that Wolbachia infection significantly regulated 91 miRNAs in females and 20 miRNAs in males, with an overall suppression of miRNAs in Wolbachia-infected libraries. A comparison of the miRNA and mRNA data predicted that the differentially expressed miRNAs negatively regulated 90 mRNAs in females and 9 mRNAs in males. An analysis of target genes showed that Wolbachia-responsive miRNAs regulated genes with function in sphingolipid metabolism, lysosome function, apoptosis and lipid transporting in both sexes, as well as reproduction in females.

CONCLUSION: Comparisons of the miRNA and mRNA data can help to identify miRNAs and miRNA target genes involving in Wolbachia-host interactions. The molecular targets identified in this study should be useful in further functional studies.}, } @article {pmid25512495, year = {2014}, author = {Mayoral, JG and Hussain, M and Joubert, DA and Iturbe-Ormaetxe, I and O'Neill, SL and Asgari, S}, title = {Wolbachia small noncoding RNAs and their role in cross-kingdom communications.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {111}, number = {52}, pages = {18721-18726}, pmid = {25512495}, issn = {1091-6490}, mesh = {Aedes/*microbiology ; Animals ; Drosophila melanogaster ; RNA, Bacterial/genetics/*metabolism ; RNA, Small Untranslated/genetics/*metabolism ; Wolbachia/genetics/*metabolism ; }, abstract = {In prokaryotes, small noncoding RNAs (snRNAs) of 50-500 nt are produced that are important in bacterial virulence and response to environmental stimuli. Here, we identified and characterized snRNAs from the endosymbiotic bacteria, Wolbachia, which are widespread in invertebrates and cause reproductive manipulations. Most importantly, some strains of Wolbachia inhibit replication of several vector-borne pathogens in insects. We demonstrate that two abundant snRNAs, WsnRNA-46 and WsnRNA-49, are expressed in Wolbachia from noncoding RNA transcripts that contain precursors with stem-loop structures. WsnRNAs were detected in Aedes aegypti mosquitoes infected with the wMelPop-CLA strain of Wolbachia and in Drosophila melanogaster and Drosophila simulans infected with wMelPop and wAu strains, respectively, indicating that the WsnRNAs are conserved across species and strains. In addition, we show that the WsnRNAs may potentially regulate host genes and Wolbachia genes. Our findings provide evidence for the production of functional snRNAs by Wolbachia that play roles in cross-kingdom communication between the endosymbiont and the host.}, } @article {pmid25502564, year = {2014}, author = {Segoli, M and Hoffmann, AA and Lloyd, J and Omodei, GJ and Ritchie, SA}, title = {The effect of virus-blocking Wolbachia on male competitiveness of the dengue vector mosquito, Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {12}, pages = {e3294}, pmid = {25502564}, issn = {1935-2735}, mesh = {Aedes/*microbiology/physiology/virology ; Animals ; Competitive Behavior/*physiology ; Dengue/microbiology/prevention & control/transmission ; Dengue Virus/*physiology ; Female ; Male ; Sexual Behavior, Animal/physiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The bacterial endosymbiont Wolbachia blocks the transmission of dengue virus by its vector mosquito Aedes aegypti, and is currently being evaluated for control of dengue outbreaks. Wolbachia induces cytoplasmic incompatibility (CI) that results in the developmental failure of offspring in the cross between Wolbachia-infected males and uninfected females. This increases the relative success of infected females in the population, thereby enhancing the spread of the beneficial bacterium. However, Wolbachia spread via CI will only be feasible if infected males are sufficiently competitive in obtaining a mate under field conditions. We tested the effect of Wolbachia on the competitiveness of A. aegypti males under semi-field conditions.

In a series of experiments we exposed uninfected females to Wolbachia-infected and uninfected males simultaneously. We scored the competitiveness of infected males according to the proportion of females producing non-viable eggs due to incompatibility. We found that infected males were equally successful to uninfected males in securing a mate within experimental tents and semi-field cages. This was true for males infected by the benign wMel Wolbachia strain, but also for males infected by the virulent wMelPop (popcorn) strain. By manipulating male size we found that larger males had a higher success than smaller underfed males in the semi-field cages, regardless of their infection status.

CONCLUSIONS/SIGNIFICANCE: The results indicate that Wolbachia infection does not reduce the competitiveness of A. aegypti males. Moreover, the body size effect suggests a potential advantage for lab-reared Wolbachia-males during a field release episode, due to their better nutrition and larger size. This may promote Wolbachia spread via CI in wild mosquito populations and underscores its potential use for disease control.}, } @article {pmid25502325, year = {2014}, author = {Touret, F and Guiguen, F and Greenland, T and Terzian, C}, title = {In between: gypsy in Drosophila melanogaster reveals new insights into endogenous retrovirus evolution.}, journal = {Viruses}, volume = {6}, number = {12}, pages = {4914-4925}, pmid = {25502325}, issn = {1999-4915}, mesh = {Animals ; Drosophila melanogaster/genetics/*virology ; Endogenous Retroviruses/*genetics/physiology ; *Evolution, Molecular ; Retroelements ; Retroviridae/*genetics/physiology ; }, abstract = {Retroviruses are RNA viruses that are able to synthesize a DNA copy of their genome and insert it into a chromosome of the host cell. Sequencing of different eukaryote genomes has revealed the presence of many such endogenous retroviral sequences. The mechanisms by which these retroviral sequences have colonized the genome are still unknown, and the endogenous retrovirus gypsy of Drosophila melanogaster is a powerful experimental model for deciphering this process in vivo. Gypsy is expressed in a layer of somatic cells, and then transferred into the oocyte by an unknown mechanism. This critical step is the start of the endogenization process. Moreover gypsy has been shown to have infectious properties, probably due to its envelope gene acquired from a baculovirus. Recently we have also shown that gypsy maternal transmission is reduced in the presence of the endosymbiotic bacterium Wolbachia. These studies demonstrate that gypsy is a unique and powerful model for understanding the endogenization of retroviruses.}, } @article {pmid25499047, year = {2015}, author = {Sakamoto, Y and Hirai, N and Tanikawa, T and Yago, M and Ishii, M}, title = {Population genetic structure and Wolbachia infection in an endangered butterfly, Zizina emelina (Lepidoptera, Lycaenidae), in Japan.}, journal = {Bulletin of entomological research}, volume = {105}, number = {2}, pages = {152-165}, doi = {10.1017/S0007485314000819}, pmid = {25499047}, issn = {1475-2670}, mesh = {Animals ; Butterflies/*genetics/*microbiology ; DNA, Mitochondrial/*chemistry ; Endangered Species ; Female ; Genetic Variation ; Host-Pathogen Interactions ; Japan ; Male ; Phylogeny ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Zizina emelina (de l'Orza) is listed on Japan's Red Data List as an endangered species because of loss of its principal food plant and habitat. We compared parts of the mitochondrial and nuclear genes of this species to investigate the level of genetic differentiation among the 14 extant populations. We also examined infection of the butterfly with the bacterium Wolbachia to clarify the bacterium's effects on the host population's genetic structure. Mitochondrial and nuclear DNA analyses revealed that haplotype composition differed significantly among most of the populations, and the fixation index F ST was positively correlated with geographic distance. In addition, we found three strains of Wolbachia, one of which was a male killer; these strains were prevalent in several populations. There was linkage between some host mitochondrial haplotypes and the three Wolbachia strains, although no significant differences were found in a comparison of host mitochondrial genetic diversity with nuclear genetic diversity in Wolbachia-infected or -uninfected populations. These genetic analyses and Wolbachia infection findings show that Z. emelina has little migratory activity and that little gene flow occurs among the current populations.}, } @article {pmid25496002, year = {2014}, author = {Klasson, L and Kumar, N and Bromley, R and Sieber, K and Flowers, M and Ott, SH and Tallon, LJ and Andersson, SG and Dunning Hotopp, JC}, title = {Extensive duplication of the Wolbachia DNA in chromosome four of Drosophila ananassae.}, journal = {BMC genomics}, volume = {15}, number = {1}, pages = {1097}, pmid = {25496002}, issn = {1471-2164}, support = {DP2 OD007372/OD/NIH HHS/United States ; T32 DK067872/DK/NIDDK NIH HHS/United States ; 1-DP2-OD007372/OD/NIH HHS/United States ; }, mesh = {Animals ; Chromosome Mapping ; *DNA Replication ; DNA, Bacterial/*biosynthesis/*genetics ; Drosophila/*genetics/*microbiology ; Female ; Gene Dosage ; *Gene Transfer, Horizontal ; Genome, Insect/genetics ; Heterozygote ; Male ; Polytene Chromosomes/genetics ; Sequence Analysis, DNA ; Species Specificity ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Lateral gene transfer (LGT) from bacterial Wolbachia endosymbionts has been detected in ~20% of arthropod and nematode genome sequencing projects. Many of these transfers are large and contain a substantial part of the Wolbachia genome.

RESULTS: Here, we re-sequenced three D. ananassae genomes from Asia and the Pacific that contain large LGTs from Wolbachia. We find that multiple copies of the Wolbachia genome are transferred to the Drosophila nuclear genome in all three lines. In the D. ananassae line from Indonesia, the copies of Wolbachia DNA in the nuclear genome are nearly identical in size and sequence yielding an even coverage of mapped reads over the Wolbachia genome. In contrast, the D. ananassae lines from Hawaii and India show an uneven coverage of mapped reads over the Wolbachia genome suggesting that different parts of these LGTs are present in different copy numbers. In the Hawaii line, we find that this LGT is underrepresented in third instar larvae indicative of being heterochromatic. Fluorescence in situ hybridization of mitotic chromosomes confirms that the LGT in the Hawaii line is heterochromatic and represents ~20% of the sequence on chromosome 4 (dot chromosome, Muller element F).

CONCLUSIONS: This collection of related lines contain large lateral gene transfers composed of multiple Wolbachia genomes that constitute >2% of the D. ananassae genome (~5 Mbp) and partially explain the abnormally large size of chromosome 4 in D. ananassae.}, } @article {pmid25482270, year = {2015}, author = {Atyame, CM and Labbé, P and Rousset, F and Beji, M and Makoundou, P and Duron, O and Dumas, E and Pasteur, N and Bouattour, A and Fort, P and Weill, M}, title = {Stable coexistence of incompatible Wolbachia along a narrow contact zone in mosquito field populations.}, journal = {Molecular ecology}, volume = {24}, number = {2}, pages = {508-521}, doi = {10.1111/mec.13035}, pmid = {25482270}, issn = {1365-294X}, mesh = {Animals ; Bacterial Typing Techniques ; Crosses, Genetic ; Culex/*microbiology ; Female ; *Genetics, Population ; Male ; Microsatellite Repeats ; Reproduction ; Tunisia ; Wolbachia/classification/*genetics ; }, abstract = {In arthropods, the intracellular bacteria Wolbachia often induce cytoplasmic incompatibility (CI) between sperm and egg, which causes conditional embryonic death and promotes the spatial spread of Wolbachia infections into host populations. The ability of Wolbachia to spread in natural populations through CI has attracted attention for using these bacteria in vector-borne disease control. The dynamics of incompatible Wolbachia infections have been deeply investigated theoretically, whereas in natural populations, there are only few examples described, especially among incompatible infected hosts. Here, we have surveyed the distribution of two molecular Wolbachia strains (wPip11 and wPip31) infecting the mosquito Culex pipiens in Tunisia. We delineated a clear spatial structure of both infections, with a sharp contact zone separating their distribution areas. Crossing experiments with isofemale lines from different localities showed three crossing types: wPip11-infected males always sterilize wPip31-infected females; however, while most wPip31-infected males were compatible with wPip11-infected females, a few completely sterilize them. The wPip11 strain was thus expected to spread, but temporal dynamics over 7 years of monitoring shows the stability of the contact zone. We examined which factors may contribute to the observed stability, both theoretically and empirically. Population cage experiments, field samples and modelling did not support significant impacts of local adaptation or assortative mating on the stability of wPip infection structure. By contrast, low dispersal probability and metapopulation dynamics in the host Cx. pipiens probably play major roles. This study highlights the need of understanding CI dynamics in natural populations to design effective and sustainable Wolbachia-based control strategies.}, } @article {pmid25474676, year = {2014}, author = {Hornett, EA and Moran, B and Reynolds, LA and Charlat, S and Tazzyman, S and Wedell, N and Jiggins, CD and Hurst, GD}, title = {The evolution of sex ratio distorter suppression affects a 25 cM genomic region in the butterfly Hypolimnas bolina.}, journal = {PLoS genetics}, volume = {10}, number = {12}, pages = {e1004822}, pmid = {25474676}, issn = {1553-7404}, mesh = {Animals ; Butterflies/*genetics ; *Evolution, Molecular ; Female ; Genetic Variation ; Genome, Insect/*genetics ; Gram-Negative Bacterial Infections/genetics/veterinary ; Host-Pathogen Interactions/*genetics ; Linkage Disequilibrium ; Male ; Molecular Sequence Data ; Samoa ; *Sex Ratio ; *Suppression, Genetic ; Wolbachia/genetics/pathogenicity ; }, abstract = {Symbionts that distort their host's sex ratio by favouring the production and survival of females are common in arthropods. Their presence produces intense Fisherian selection to return the sex ratio to parity, typified by the rapid spread of host 'suppressor' loci that restore male survival/development. In this study, we investigated the genomic impact of a selective event of this kind in the butterfly Hypolimnas bolina. Through linkage mapping, we first identified a genomic region that was necessary for males to survive Wolbachia-induced male-killing. We then investigated the genomic impact of the rapid spread of suppression, which converted the Samoan population of this butterfly from a 100:1 female-biased sex ratio in 2001 to a 1:1 sex ratio by 2006. Models of this process revealed the potential for a chromosome-wide effect. To measure the impact of this episode of selection directly, the pattern of genetic variation before and after the spread of suppression was compared. Changes in allele frequencies were observed over a 25 cM region surrounding the suppressor locus, with a reduction in overall diversity observed at loci that co-segregate with the suppressor. These changes exceeded those expected from drift and occurred alongside the generation of linkage disequilibrium. The presence of novel allelic variants in 2006 suggests that the suppressor was likely to have been introduced via immigration rather than through de novo mutation. In addition, further sampling in 2010 indicated that many of the introduced variants were lost or had declined in frequency since 2006. We hypothesize that this loss may have resulted from a period of purifying selection, removing deleterious material that introgressed during the initial sweep. Our observations of the impact of suppression of sex ratio distorting activity reveal a very wide genomic imprint, reflecting its status as one of the strongest selective forces in nature.}, } @article {pmid25473839, year = {2014}, author = {Ferreira, ÁG and Naylor, H and Esteves, SS and Pais, IS and Martins, NE and Teixeira, L}, title = {The Toll-dorsal pathway is required for resistance to viral oral infection in Drosophila.}, journal = {PLoS pathogens}, volume = {10}, number = {12}, pages = {e1004507}, pmid = {25473839}, issn = {1553-7374}, support = {BB/E005470/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Drosophila Proteins/genetics/*immunology ; Drosophila melanogaster ; *Immunity, Innate ; *Mutation ; Nuclear Proteins/genetics/*immunology ; Phosphoproteins/genetics/*immunology ; Picornaviridae/*immunology ; Signal Transduction/genetics/*immunology ; Toll-Like Receptors/genetics/*immunology ; Transcription Factors/genetics/*immunology ; }, abstract = {Pathogen entry route can have a strong impact on the result of microbial infections in different hosts, including insects. Drosophila melanogaster has been a successful model system to study the immune response to systemic viral infection. Here we investigate the role of the Toll pathway in resistance to oral viral infection in D. melanogaster. We show that several Toll pathway components, including Spätzle, Toll, Pelle and the NF-kB-like transcription factor Dorsal, are required to resist oral infection with Drosophila C virus. Furthermore, in the fat body Dorsal is translocated from the cytoplasm to the nucleus and a Toll pathway target gene reporter is upregulated in response to Drosophila C Virus infection. This pathway also mediates resistance to several other RNA viruses (Cricket paralysis virus, Flock House virus, and Nora virus). Compared with control, viral titres are highly increased in Toll pathway mutants. The role of the Toll pathway in resistance to viruses in D. melanogaster is restricted to oral infection since we do not observe a phenotype associated with systemic infection. We also show that Wolbachia and other Drosophila-associated microbiota do not interact with the Toll pathway-mediated resistance to oral infection. We therefore identify the Toll pathway as a new general inducible pathway that mediates strong resistance to viruses with a route-specific role. These results contribute to a better understanding of viral oral infection resistance in insects, which is particularly relevant in the context of transmission of arboviruses by insect vectors.}, } @article {pmid25470996, year = {2014}, author = {Raphael, KA and Shearman, DC and Gilchrist, AS and Sved, JA and Morrow, JL and Sherwin, WB and Riegler, M and Frommer, M}, title = {Australian endemic pest tephritids: genetic, molecular and microbial tools for improved Sterile Insect Technique.}, journal = {BMC genetics}, volume = {15 Suppl 2}, number = {Suppl 2}, pages = {S9}, pmid = {25470996}, issn = {1471-2156}, mesh = {Animals ; *Animals, Genetically Modified ; Australia ; *Biotechnology ; Diptera/*genetics ; Female ; Gene Expression Profiling ; Genetic Markers ; Genome, Insect ; Infertility/*genetics ; Male ; Sex Factors ; }, abstract = {Among Australian endemic tephritid fruit flies, the sibling species Bactrocera tryoni and Bactrocera neohumeralis have been serious horticultural pests since the introduction of horticulture in the nineteenth century. More recently, Bactrocera jarvisi has also been declared a pest in northern Australia. After several decades of genetic research there is now a range of classical and molecular genetic tools that can be used to develop improved Sterile Insect Technique (SIT) strains for control of these pests. Four-way crossing strategies have the potential to overcome the problem of inbreeding in mass-reared strains of B. tryoni. The ability to produce hybrids between B. tryoni and the other two species in the laboratory has proved useful for the development of genetically marked strains. The identification of Y-chromosome markers in B. jarvisi means that male and female embryos can be distinguished in any strain that carries a B. jarvisi Y chromosome. This has enabled the study of homologues of the sex-determination genes during development of B jarvisi and B. tryoni, which is necessary for the generation of genetic-sexing strains. Germ-line transformation has been established and a draft genome sequence for B. tryoni released. Transcriptomes from various species, tissues and developmental stages, to aid in identification of manipulation targets for improving SIT, have been assembled and are in the pipeline. Broad analyses of the microbiome have revealed a metagenome that is highly variable within and across species and defined by the environment. More specific analyses detected Wolbachia at low prevalence in the tropics but absent in temperate regions, suggesting a possible role for this endosymbiont in future control strategies.}, } @article {pmid25458121, year = {2014}, author = {McCall, JW and Kramer, L and Genchi, C and Guerrero, J and Dzimianski, MT and Mansour, A and McCall, SD and Carson, B}, title = {Effects of doxycycline on heartworm embryogenesis, transmission, circulating microfilaria, and adult worms in microfilaremic dogs.}, journal = {Veterinary parasitology}, volume = {206}, number = {1-2}, pages = {5-13}, doi = {10.1016/j.vetpar.2014.09.023}, pmid = {25458121}, issn = {1873-2550}, mesh = {Animals ; Antigens, Helminth/blood ; Culicidae/parasitology ; Dirofilaria immitis/*drug effects/embryology ; Dirofilariasis/*drug therapy/transmission ; Dog Diseases/*drug therapy/transmission ; Dogs ; Doxycycline/*pharmacology/*therapeutic use ; Embryonic Development/drug effects ; Microfilariae/drug effects ; }, abstract = {Tetracycline treatment of animals or humans infected with filariae that harbor Wolbachia endosymbionts blocks further embryogenesis, and existing microfilariae gradually die. This treatment also kills developing larvae and has a slow-kill effect on adult filariae, all presumably due to elimination of the Wolbachia. Also, Dirofilaria immitis microfilariae in blood collected from dogs up to 25 days after the last dose of doxycycline developed to infective L3 that were normal in appearance and motility in mosquitoes but did not continue to develop or migrate normally after subcutaneous (SC) injection into dogs. The present study was designed to determine whether heartworm microfilariae collected at later times after treatment would regain the ability to continue normal development in a dog. The study also was expected to yield valuable data on the effects of treatment on microfilariae and antigen levels and adult worms. The study was conducted in 16 dogs as two separate replicates at different times. A total of five dogs (two in Replicate A and three in Replicate B) infected either by SC injection of L3 or intravenous transplantation of adult heartworms were given doxycycline orally at 10mg/kg twice daily for 30 days, with three untreated controls. Microfilarial counts in the five treated dogs gradually declined during the 12-13 months after treatment initiation. Two dogs were amicrofilaremic before necropsy and three had 13 or fewer microfilariae/ml. Only one treated dog was negative for heartworm antigen before necropsy. Overall, treated dogs generally had fewer live adult heartworms than controls, and most of their live worms were moribund. All three control dogs remained positive for microfilariae and antigen and had many live worms. L3 from mosquitoes fed on blood collected 73-77 or 161-164 days after initiation of doxycycline treatments were injected SC into five dogs. None of the dogs injected with L3 from mosquitoes fed on blood from doxycycline-treated dogs were ever positive for microfilariae or antigen, and none had worms at necropsy; three control dogs were positive for microfilariae and antigen and had many live worms. These data indicate that doxycycline treatment of microfilaremic dogs gradually reduces numbers of microfilariae and blocks further transmission of heartworms. This latter effect should be highly effective in reducing the rate of selection of heartworms with genes that confer resistance to macrocyclic lactone preventives and microfilaricides. The data also suggest that doxycycline has a slow-kill effect on adult heartworms.}, } @article {pmid25452279, year = {2015}, author = {Newton, IL and Sheehan, KB}, title = {Passage of Wolbachia pipientis through mutant drosophila melanogaster induces phenotypic and genomic changes.}, journal = {Applied and environmental microbiology}, volume = {81}, number = {3}, pages = {1032-1037}, pmid = {25452279}, issn = {1098-5336}, support = {P40 OD018537/OD/NIH HHS/United States ; P40OD018537/OD/NIH HHS/United States ; }, mesh = {*Adaptation, Biological ; Animals ; Bacterial Outer Membrane Proteins/genetics/metabolism ; Drosophila melanogaster/genetics/*microbiology ; Gastrointestinal Tract/microbiology ; Genome, Bacterial ; Mutation ; Sequence Analysis, DNA ; Wolbachia/*genetics/isolation & purification/*physiology ; }, abstract = {Wolbachia pipientis is a nearly ubiquitous, maternally transmitted bacterium that infects the germ line of insect hosts. Estimates are that Wolbachia infects 40 to 60% of insect species on the planet, making it one of the most prevalent infections on Earth. However, we know surprisingly little about the molecular mechanisms used by Wolbachia to infect its hosts. We passaged Wolbachia through normally restrictive Drosophila melanogaster hosts, bottlenecking Wolbachia through stochastic segregation while simultaneously selecting for mutants that could recolonize these previously restrictive hosts. Here, we show that Wolbachia alters its behavior when passaged through heterozygous mutant flies. After only three generations, Wolbachia was able to colonize the previously restrictive hosts at control titers. Additionally, the Wolbachia organisms passaged through heterozygous mutant D. melanogaster alter their pattern of tissue-specific Wsp protein production, suggesting a behavioral response to the host genotype. Using whole-genome resequencing, we identified the mutations accumulated by these lineages of Wolbachia and confirmed the existence and persistence of the mutations through clone library Sanger sequencing. Our results suggest that Wolbachia can quickly adapt to new host contexts, with genomic mutants arising after only two generations.}, } @article {pmid25445614, year = {2014}, author = {Tamarozzi, F and Wright, HL and Thomas, HB and Edwards, SW and Taylor, MJ}, title = {A lack of confirmation with alternative assays questions the validity of IL-17A expression in human neutrophils using immunohistochemistry.}, journal = {Immunology letters}, volume = {162}, number = {2 Pt B}, pages = {194-198}, doi = {10.1016/j.imlet.2014.10.025}, pmid = {25445614}, issn = {1879-0542}, support = {19437//Arthritis Research UK/United Kingdom ; BB/H016163/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Antibodies, Helminth/*chemistry/immunology ; Female ; Gene Expression Regulation/drug effects/immunology ; Humans ; Immunohistochemistry/methods ; Interleukin-17/*immunology ; Male ; Neutrophils/*immunology/pathology ; Onchocerca volvulus/*immunology ; Onchocerciasis/drug therapy/*immunology/pathology ; }, abstract = {We identified IL-17A-positive neutrophils in Wolbachia-positive Onchocerca volvulus nodules using an antibody that has previously reported IL-17A-positive neutrophils in several inflammatory conditions. However, we could not detect IL-17A using a range of alternative assays. Our data question the IL-17A antibody specificity and the ability of human neutrophils to express IL-17A.}, } @article {pmid25433394, year = {2014}, author = {Luck, AN and Evans, CC and Riggs, MD and Foster, JM and Moorhead, AR and Slatko, BE and Michalski, ML}, title = {Concurrent transcriptional profiling of Dirofilaria immitis and its Wolbachia endosymbiont throughout the nematode life cycle reveals coordinated gene expression.}, journal = {BMC genomics}, volume = {15}, number = {1}, pages = {1041}, pmid = {25433394}, issn = {1471-2164}, mesh = {Animals ; Dirofilaria immitis/*genetics/pathogenicity ; Dirofilariasis/genetics/parasitology ; Dogs ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Life Cycle Stages/genetics ; Male ; Microfilariae/*genetics/parasitology ; Symbiosis/*genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {BACKGROUND: Dirofilaria immitis, or canine heartworm, is a filarial nematode parasite that infects dogs and other mammals worldwide. Current disease control relies on regular administration of anthelmintic preventives, however, relatively poor compliance and evidence of developing drug resistance could warrant alternative measures against D. immitis and related human filarial infections be taken. As with many other filarial nematodes, D. immitis contains Wolbachia, an obligate bacterial endosymbiont thought to be involved in providing certain critical metabolites to the nematode. Correlations between nematode and Wolbachia transcriptomes during development have not been examined. Therefore, we detailed the developmental transcriptome of both D. immitis and its Wolbachia (wDi) in order to gain a better understanding of parasite-endosymbiont interactions throughout the nematode life cycle.

RESULTS: Over 215 million single-end 50 bp reads were generated from total RNA from D. immitis adult males and females, microfilariae (mf) and third and fourth-stage larvae (L3 and L4). We critically evaluated the transcriptomes of the various life cycle stages to reveal sex-biased transcriptional patterns, as well as transcriptional differences between larval stages that may be involved in larval maturation. Hierarchical clustering revealed both D. immitis and wDi transcriptional activity in the L3 stage is clearly distinct from other life cycle stages. Interestingly, a large proportion of both D. immitis and wDi genes display microfilarial-biased transcriptional patterns. Concurrent transcriptome sequencing identified potential molecular interactions between parasite and endosymbiont that are more prominent during certain life cycle stages. In support of metabolite provisioning between filarial nematodes and Wolbachia, the synthesis of the critical metabolite, heme, by wDi appears to be synchronized in a stage-specific manner (mf-specific) with the production of heme-binding proteins in D. immitis.

CONCLUSIONS: Our integrated transcriptomic study has highlighted interesting correlations between Wolbachia and D. immitis transcription throughout the life cycle and provided a resource that may be used for the development of novel intervention strategies, not only for the treatment and prevention of D. immitis infections, but of other closely related human parasites as well.}, } @article {pmid25417905, year = {2014}, author = {Favia, G}, title = {Symbionts conferring resistance to viruses in insects.}, journal = {Pathogens and global health}, volume = {108}, number = {7}, pages = {303-304}, pmid = {25417905}, issn = {2047-7732}, mesh = {Animals ; Drosophila/*growth & development/*immunology ; Female ; Host-Pathogen Interactions/*immunology ; Insect Viruses/*pathogenicity ; Male ; Symbiosis/*immunology ; Virus Diseases/*immunology ; Wolbachia/*physiology ; }, } @article {pmid25405506, year = {2014}, author = {Avtzis, DN and Doudoumis, V and Bourtzis, K}, title = {Wolbachia infections and mitochondrial diversity of two chestnut feeding Cydia species.}, journal = {PloS one}, volume = {9}, number = {11}, pages = {e112795}, pmid = {25405506}, issn = {1932-6203}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; *Host Specificity ; Moths/*genetics/microbiology ; Phylogeny ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Cydia splendana and C. fagiglandana are two closely related chestnut feeding lepidopteran moth species. In this study, we surveyed the bacterial endosymbiont Wolbachia in these two species. Infection rates were 31% in C. splendana and 77% in C. fagiglandana. MLST analysis showed that these two species are infected with two quite diverse Wolbachia strains. C. splendana with Sequence Type (ST) 409 from the A-supergroup and C. fagiglandana with ST 150 from the B-supergroup. One individual of C. splendana was infected with ST 150, indicating horizontal transfer between these sister species. The mitochondrial DNA of the two Cydia species showed a significantly different mtDNA diversity, which was inversely proportional to their infection rates.}, } @article {pmid25394184, year = {2016}, author = {Strunov, A and Kiseleva, E}, title = {Drosophila melanogaster brain invasion: pathogenic Wolbachia in central nervous system of the fly.}, journal = {Insect science}, volume = {23}, number = {2}, pages = {253-264}, doi = {10.1111/1744-7917.12187}, pmid = {25394184}, issn = {1744-7917}, mesh = {Animals ; Brain/microbiology/ultrastructure ; Drosophila melanogaster/*microbiology/ultrastructure ; Female ; Microscopy, Electron, Transmission ; Neuroglia/microbiology ; Neurons/microbiology ; Wolbachia/*physiology ; }, abstract = {The pathogenic Wolbachia strain wMelPop rapidly over-replicates in the brain, muscles, and retina of Drosophila melanogaster, causing severe tissue degeneration and premature death of the host. The unique features of this endosymbiont make it an excellent tool to be used for biological control of insects, pests, and vectors of human diseases. To follow the dynamics of bacterial morphology and titer in the nerve cells we used transmission electron microscopy of 3-d-old female brains. The neurons and glial cells from central brain of the fly had different Wolbachia titers ranging from single bacteria to large accumulations, tearing cell apart and invading extracellular space. The neuropile regions of the brain were free of wMelPop. Wolbachia tightly interacted with host cell organelles and underwent several morphological changes in nerve cells. Based on different morphological types of bacteria described we propose for the first time a scheme of wMelPop dynamics within the somatic tissue of the host.}, } @article {pmid25387458, year = {2014}, author = {Montoya-Alonso, JA and Carretón, E and García-Guasch, L and Expósito, J and Armario, B and Morchón, R and Simón, F}, title = {First epidemiological report of feline heartworm infection in the Barcelona metropolitan area (Spain).}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {506}, pmid = {25387458}, issn = {1756-3305}, mesh = {Animals ; Cat Diseases/blood/diagnosis/*epidemiology/parasitology ; Cats ; Cities/epidemiology ; Dirofilaria immitis/immunology/*isolation & purification ; Dirofilariasis/blood/diagnosis/*epidemiology/parasitology ; Dog Diseases/blood/epidemiology/parasitology ; Dogs ; Female ; Male ; Spain/epidemiology ; }, abstract = {BACKGROUND: The metropolitan area of Barcelona is the most densely populated metropolitan area on the Mediterranean coast. Several studies have reported the presence of canine heartworm disease in this region; however, there are no published epidemiological data regarding feline heartworm in this region and the prevalence in this species remains unknown.

METHODS: Serum samples from 758 cats living in the metropolitan area of Barcelona (Spain) were collected between 2012 and 2013. To establish the seroprevalence of heartworm infection in cats, serological techniques for anti-D.immitis and anti-Wolbachia antibody detection were used while a commercial ELISA test kit was used to detect circulating D.immitis antigens.

RESULTS: Of these samples, 11.47% were positive to D.immitis and Wolbachia surface protein antibodies and 0.26% were positive to D.immitis antigens. The higher antibody seroprevalences were found in the areas that follow the courses of the rivers Llobregat and Anoia (Baix Llobregat 11.5%, Vallés Occidental 13.2%; Barcelonés 11.7%) where humidity and vegetation favour the development of the mosquito vectors. High antibody seroprevalences were also found in the urban areas (Barcelona city 13.1%; Sabadell 15.5%), which demonstrates that city cats are also at risk from D.immitis infection.

CONCLUSIONS: Generally, in Spain cats do not receive prophylactic treatment and therefore the risk of infection is higher in this species than in dogs. Adequate prophylactic plans should be implemented in the feline population. This is the first epidemiologic study on feline heartworm infection to be carried out in continental Spain.}, } @article {pmid25373153, year = {2014}, author = {Hamm, CA and Handley, CA and Pike, A and Forister, ML and Fordyce, JA and Nice, CC}, title = {Wolbachia infection and Lepidoptera of conservation concern.}, journal = {Journal of insect science (Online)}, volume = {14}, number = {}, pages = {6}, pmid = {25373153}, issn = {1536-2442}, mesh = {Animals ; *Conservation of Natural Resources ; DNA, Bacterial/genetics ; Female ; Lepidoptera/*microbiology ; Male ; Polymerase Chain Reaction ; United States ; Wolbachia/genetics/*physiology ; }, abstract = {Conservation of at-risk species requires multi-faceted and carefully-considered management approaches to be successful. For arthropods, the presence of endosymbiotic bacteria, such as Wolbachia (Rickettsiales: Rickettsiaceae), may complicate management plans and exacerbate the challenges faced by conservation managers. Wolbachia poses a substantial and underappreciated threat to the conservation of arthropods because infection may induce a number of phenotypic effects, most of which are considered deleterious to the host population. In this study, the prevalence of Wolbachia infection in lepidopteran species of conservation concern was examined. Using standard molecular techniques, 22 species of Lepidoptera were screened, of which 19 were infected with Wolbachia. This rate is comparable to that observed in insects as a whole. However, this is likely an underestimate because geographic sampling was not extensive and may not have included infected segments of the species' ranges. Wolbachia infections may be particularly problematic for conservation management plans that incorporate captive propagation or translocation. Inadvertent introduction of Wolbachia into uninfected populations or introduction of a new strain may put these populations at greater risk for extinction. Further sampling to investigate the geographic extent of Wolbachia infections within species of conservation concern and experiments designed to determine the nature of the infection phenotype(s) are necessary to manage the potential threat of infection.}, } @article {pmid25368078, year = {2014}, author = {Pistone, D and Bione, A and Epis, S and Pajoro, M and Gaiarsa, S and Bandi, C and Sassera, D}, title = {Presence of Wolbachia in three hymenopteran species: Diprion pini (Hymenoptera: Diprionidae), Neodiprion sertifer (Hymenoptera: Diprionidae), and Dahlbominus fuscipennis (Hymenoptera: Eulophidae).}, journal = {Journal of insect science (Online)}, volume = {14}, number = {}, pages = {147}, pmid = {25368078}, issn = {1536-2442}, mesh = {Animals ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Female ; Hot Temperature ; Hymenoptera/anatomy & histology/*microbiology/*physiology ; Italy ; Male ; Mosaicism ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sex Characteristics ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Sawflies are important pests of various plant species. Diprion pini (L.) and Neodiprion sertifer (Geoffroy) (Hymenoptera: Diprionidae) are two of the most important sawfly pests in Italy, and both species are parasitized by the hymenopteran parasitoid Dahlbominus fuscipennis (Zetterstedt). Bacterial endosymbionts are currently studied for their high potential in strategies of biocontrol in a number of insect species. In this study, we investigated the presence of symbiotic bacteria (Wolbachia and Cardinium) in the three species of hymenoptera mentioned earlier, both in wild and laboratory populations. Although all samples were negative for the presence of Cardinium, 100% prevalence for Wolbachia was detected, as all examined individuals resulted to be PCR positive. Furthermore, 16S rDNA and ftsZ gene sequencing indicated that all individuals from the three hymenopteran species are infected by a single Wolbachia strain. Additionally, we report the presence of gynandromorphic individuals in D. pini, both in wild and laboratory-reared populations. Heat treatments on D. pini colonies removed the Wolbachia symbionts, but they also prevented the development of adults.}, } @article {pmid25365330, year = {2014}, author = {Kollenberg, M and Winter, S and Götz, M}, title = {Quantification and localization of Watermelon chlorotic stunt virus and Tomato yellow leaf curl virus (Geminiviridae) in populations of Bemisia tabaci (Hemiptera, Aleyrodidae) with differential virus transmission characteristics.}, journal = {PloS one}, volume = {9}, number = {11}, pages = {e111968}, pmid = {25365330}, issn = {1932-6203}, mesh = {Animals ; *Geminiviridae ; Hemiptera/*virology ; Insect Vectors/*virology ; Intestines/*virology ; Plant Diseases/*virology ; Salivary Glands/*virology ; }, abstract = {Bemisia tabaci (Gennadius) is one of the economically most damaging insects to crops in tropical and subtropical regions. Severe damage is caused by feeding and more seriously by transmitting viruses. Those of the genus begomovirus (Geminiviridae) cause the most significant crop diseases and are transmitted by B. tabaci in a persistent circulative mode, a process which is largely unknown. To analyze the translocation and to identify critical determinants for transmission, two populations of B. tabaci MEAM1 were compared for transmitting Watermelon chlorotic stunt virus (WmCSV) and Tomato yellow leaf curl virus (TYLCV). Insect populations were chosen because of their high and respectively low virus transmission efficiency to compare uptake and translocation of virus through insects. Both populations harbored Rickettsia, Hamiltonella and Wolbachia in comparable ratios indicating that endosymbionts might not contribute to the different transmission rates. Quantification by qPCR revealed that WmCSV uptake and virus concentrations in midguts and primary salivary glands were generally higher than TYLCV due to higher virus contents of the source plants. Both viruses accumulated higher in insects from the efficiently compared to the poorly transmitting population. In the latter, virus translocation into the hemolymph was delayed and virus passage was impeded with limited numbers of viruses translocated. FISH analysis confirmed these results with similar virus distribution found in excised organs of both populations. No virus accumulation was found in the midgut lumen of the poor transmitter because of a restrained virus translocation. Results suggest that the poorly transmitting population comprised insects that lacked transmission competence. Those were selected to develop a population that lacks virus transmission. Investigations with insects lacking transmission showed that virus concentrations in midguts were reduced and only negligible virus amounts were found at the primary salivary glands indicating for a missing or modified receptor responsible for virus attachment or translocation.}, } @article {pmid25354059, year = {2014}, author = {March-Rosselló, GA and Eiros-Bouza, JM}, title = {[How to fight parasitic infectious diseases with bacteria. The case of Wolbachia pipientis].}, journal = {Revista medica del Instituto Mexicano del Seguro Social}, volume = {52}, number = {6}, pages = {654-659}, pmid = {25354059}, issn = {2448-5667}, mesh = {Animals ; Arthropods/*microbiology ; Filariasis/microbiology/*prevention & control/transmission ; Humans ; Nematoda/*microbiology ; *Symbiosis ; *Wolbachia ; }, abstract = {In Nature, no individual can live in isolation; hence, living organisms are forced to interact with each other. This necessity has led many organisms to establish heterogeneous relations to enhance their ability to adapt to the environment, thus acquiring evolutionary advantages. These relationships are sometimes so intense, that on the long term the organisms may lose their individual identity. An example of these associations is the endosymbiotic ones, where eukaryote organisms generally harbor different prokaryote organisms. The endosymbiotic bacterium Wolbachia pipientis is a species described by Hertig and Wolbach in 1924. This microorganism can be isolated in a large variety of eukaryote organisms, with which it maintains different links. Until now, this species has only been described with 11 serogroups numbered from A to K within the Wolbachia genus. This work is intended to illustrate the relationship of Wolbachia pipientis with human pathogenic filaria and with arthropods, as well as to describe the implications of this bacterium in the treatment of filariasis. Finally, this work tries to describe recent studies that have targeted the use of artificially-created Wolbachia pipientis virulent strains that, once inoculated in infectious diseases-transmitting vectors, develop negative effects within them in order to, in this way, erradicate mosquito-transmitted infectious diseases for which no treatment is available at the moment or the prevention of its transmissibility has not been achieved.}, } @article {pmid25347417, year = {2014}, author = {Carvalho, GA and Vieira, JL and Haro, MM and Corrêa, AS and Ribon, AO and de Oliveira, LO and Guedes, RN}, title = {Pleiotropic impact of endosymbiont load and co-occurrence in the maize weevil Sitophilus zeamais.}, journal = {PloS one}, volume = {9}, number = {10}, pages = {e111396}, pmid = {25347417}, issn = {1932-6203}, mesh = {Animals ; Bacterial Load ; Coleoptera/*microbiology/physiology ; Locomotion ; Reproduction ; *Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Individual traits vary among and within populations, and the co-occurrence of different endosymbiont species within a host may take place under varying endosymbiont loads in each individual host. This makes the recognition of the potential impact of such endosymbiont associations in insect species difficult, particularly in insect pest species. The maize weevil, Sitophilus zeamais Motsch. (Coleoptera: Curculionidae), a key pest species of stored cereal grains, exhibits associations with two endosymbiotic bacteria: the obligatory endosymbiont SZPE ("Sitophilus zeamais Primary Endosymbiont") and the facultative endosymbiont Wolbachia. The impact of the lack of SZPE in maize weevil physiology is the impairment of nutrient acquisition and energy metabolism, while Wolbachia is an important factor in reproductive incompatibility. However, the role of endosymbiont load and co-occurrence in insect behavior, grain consumption, body mass and subsequent reproductive factors has not yet been explored. Here we report on the impacts of co-occurrence and varying endosymbiont loads achieved via thermal treatment and antibiotic provision via ingested water in the maize weevil. SZPE exhibited strong effects on respiration rate, grain consumption and weevil body mass, with observed effects on weevil behavior, particularly flight activity, and potential consequences for the management of this pest species. Wolbachia directly favored weevil fertility and exhibited only mild indirect effects, usually enhancing the SZPE effect. SZPE suppression delayed weevil emergence, which reduced the insect population growth rate, and the thermal inactivation of both symbionts prevented insect reproduction. Such findings are likely important for strain divergences reported in the maize weevil and their control, aspects still deserving future attention.}, } @article {pmid25341639, year = {2014}, author = {Sutton, ER and Harris, SR and Parkhill, J and Sinkins, SP}, title = {Comparative genome analysis of Wolbachia strain wAu.}, journal = {BMC genomics}, volume = {15}, number = {1}, pages = {928}, pmid = {25341639}, issn = {1471-2164}, support = {095121//Wellcome Trust/United Kingdom ; BB/J012696/1//Biotechnology and Biological Sciences Research Council/United Kingdom ; WT095121MA//Wellcome Trust/United Kingdom ; WT098051//Wellcome Trust/United Kingdom ; }, mesh = {Bacterial Proteins/*genetics ; Evolution, Molecular ; Gene Rearrangement ; Genome, Bacterial ; Molecular Sequence Data ; Sequence Analysis, DNA ; Transcription Factors/genetics ; Wolbachia/*classification/*genetics ; }, abstract = {BACKGROUND: Wolbachia intracellular bacteria can manipulate the reproduction of their arthropod hosts, including inducing sterility between populations known as cytoplasmic incompatibility (CI). Certain strains have been identified that are unable to induce or rescue CI, including wAu from Drosophila. Genome sequencing and comparison with CI-inducing related strain wMel was undertaken in order to better understand the molecular basis of the phenotype.

RESULTS: Although the genomes were broadly similar, several rearrangements were identified, particularly in the prophage regions. Many orthologous genes contained single nucleotide polymorphisms (SNPs) between the two strains, but a subset containing major differences that would likely cause inactivation in wAu were identified, including the absence of the wMel ortholog of a gene recently identified as a CI candidate in a proteomic study. The comparative analyses also focused on a family of transcriptional regulator genes implicated in CI in previous work, and revealed numerous differences between the strains, including those that would have major effects on predicted function.

CONCLUSIONS: The study provides support for existing candidates and novel genes that may be involved in CI, and provides a basis for further functional studies to examine the molecular basis of the phenotype.}, } @article {pmid25338621, year = {2014}, author = {Halliday, A and Guimaraes, AF and Tyrer, HE and Metuge, HM and Patrick, CN and Arnaud, KO and Kwenti, TD and Forsbrook, G and Steven, A and Cook, D and Enyong, P and Wanji, S and Taylor, MJ and Turner, JD}, title = {A murine macrofilaricide pre-clinical screening model for onchocerciasis and lymphatic filariasis.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {472}, pmid = {25338621}, issn = {1756-3305}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Brugia malayi ; Elephantiasis, Filarial/*drug therapy ; Female ; Filaricides/*pharmacology ; Gerbillinae ; Male ; Menotropins ; Mice ; Mice, Inbred BALB C ; Mice, SCID ; Onchocerca ; Onchocerciasis/*drug therapy ; }, abstract = {BACKGROUND: New drugs effective against adult filariae (macrofilaricides) would accelerate the elimination of lymphatic filariasis and onchocerciasis. Anti-Onchocerca drug development is hampered by the lack of a facile model. We postulated that SCID mice could be developed as a fmacrofilaricide screening model.

METHODS: The filaricides: albendazole (ABZ), diethylcarbamazine (DEC), flubendazole (FBZ), ivermectin (IVM) and the anti-Wolbachia macrofilaricide, minocycline (MIN) were tested in Brugia malayi (Bm)-parasitized BALB/c SCID mice vs vehicle control (VC). Responses were compared to BALB/c wild type (WT). Onchocerca ochengi male worms or onchocercomata were surgically implanted into BALB/c SCID, CB.17 SCID, BALB/c WT mice or Meriones gerbils. Survival was evaluated at 7-15 days. BALB/c SCID were tested to evaluate the responsiveness of pre-clinical macrofilaricides FBZ and rifapentine (RIFAP) against male Onchocerca.

RESULTS: WT and SCID responded with >95% efficacy following ABZ or DEC treatments against Bm larvae (P < 0.0001). IVM was partially filaricidal against Bm larvae in WT and SCID (WT; 39.8%, P = 0.0356 and SCID; 56.7%, P = 0.026). SCID responded similarly to WT following IVM treatment of microfilaraemias (WT; 79%, P = 0.0194. SCID; 76%, P = 0.0473). FBZ induced a total macrofilaricidal response against adult Bm in WT and SCID (WT; P = 0.0067, SCID; P = 0.0071). MIN induced a >90% reduction in Bm Wolbachia burdens (P < 0.0001) and a blockade of microfilarial release (P = 0.0215) in SCID. Male Onchocerca survival was significantly higher in SCID vs WT mice, but not gerbils, after +15 days (60% vs 22% vs 39% P = 0.0475). Onchocercoma implants had engrafted into host tissues, with evidence of neovascularisation, after +7 days and yielded viable macro/microfilariae ex vivo. FBZ induced a macrofilaricidal effect in Onchocerca male implanted SCID at +5 weeks (FBZ; 1.67% vs VC; 43.81%, P = 0.0089). Wolbachia loads within male Onchocerca were reduced by 99% in implanted SCID receiving RIFAP for +2 weeks.

CONCLUSIONS: We have developed a 'pan-filarial' small animal research model that is sufficiently robust, with adequate capacity and throughput, to screen existing and future pre-clinical candidate macrofilaricides. Pilot data suggests a murine onchocercoma xenograft model is achievable.}, } @article {pmid25336385, year = {2015}, author = {Iwase, SI and Nakahira, K and Tuda, M and Kagoshima, K and Takagi, M}, title = {Host-plant dependent population genetics of the invading weevil Hypera postica.}, journal = {Bulletin of entomological research}, volume = {105}, number = {1}, pages = {92-100}, doi = {10.1017/S0007485314000728}, pmid = {25336385}, issn = {1475-2670}, mesh = {Animals ; Cell Nucleus/genetics ; DNA, Mitochondrial/genetics ; Electron Transport Complex IV/genetics ; Fabaceae/*physiology ; Feeding Behavior ; *Food Chain ; *Genetic Variation ; Insect Proteins/genetics ; *Introduced Species ; Japan ; Molecular Sequence Data ; Peptide Elongation Factor 1/genetics ; Polymerase Chain Reaction ; Population Density ; RNA, Transfer, Leu/genetics ; Sequence Analysis, DNA ; Species Specificity ; Weevils/genetics/microbiology/*physiology ; Wolbachia/isolation & purification ; }, abstract = {Population genetics of invading pests can be informative for understanding their ecology. In this study, we investigated population genetics of the invasive alfalfa weevil Hypera postica in Fukuoka Prefecture, Japan. We analyzed mitochondrial tRNALeu-COII, nuclear EF-1α gene fragments, and Wolbachia infection in relation to three leguminous host plants: Vicia angustifolia, Vicia villosa, and a new host Astragalus sinicus cultivated as a honey source and green manure crop. A parsimony network generated from mitochondrial gene sequences uncovered two major haplotypic groups, Western and Egyptian. In contrast to reported Wolbachia infection of the Western strain in the United States, none of our analyzed individuals were infected. The absence of Wolbachia may contribute to the stable coexistence of mitochondrial strains through inter-strain reproductive compatibility. Hypera postica genetic variants for the mitochondrial and nuclear genes were associated neither with host plant species nor with two geographic regions (Hisayama and Kama) within Fukuoka. Mitochondrial haplogroups were incongruent with nuclear genetic variants. Genetic diversity at the nuclear locus was the highest for the populations feeding on V. angustifolia. The nuclear data for A. sinicus-feeding populations indicated past sudden population growth and extended Bayesian skyline plot analysis based on the mitochondrial and nuclear data showed that the growth of A. sinicus-feeding population took place within the past 1000 years. These results suggest a shorter history of A. sinicus as a host plant compared with V. angustifolia and a recent rapid growth of H. postica population using the new host A. sinicus.}, } @article {pmid25329055, year = {2014}, author = {Gill, AC and Darby, AC and Makepeace, BL}, title = {Iron necessity: the secret of Wolbachia's success?.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {10}, pages = {e3224}, pmid = {25329055}, issn = {1935-2735}, mesh = {Animals ; Apoptosis ; Arthropods/*microbiology ; Autophagy ; Heme/biosynthesis ; Homeostasis ; Humans ; Iron/*metabolism ; Oxidative Stress ; Wolbachia/*metabolism ; }, abstract = {The bacterium Wolbachia (order Rickettsiales) is probably the world's most successful vertically-transmitted symbiont, distributed among a staggering 40% of terrestrial arthropod species. Wolbachia has great potential in vector control due to its ability to manipulate its hosts' reproduction and to impede the replication and dissemination of arboviruses and other pathogens within haematophagous arthropods. In addition, the unexpected presence of Wolbachia in filarial nematodes of medical and veterinary importance has provided an opportunity to target the adult worms of Wuchereria bancrofti, Onchocerca volvulus, and Dirofilaria immitis with safe drugs such as doxycycline. A striking feature of Wolbachia is its phenotypic plasticity between (and sometimes within) hosts, which may be underpinned by its ability to integrate itself into several key processes within eukaryotic cells: oxidative stress, autophagy, and apoptosis. Importantly, despite significant differences in the genomes of arthropod and filarial Wolbachia strains, these nexuses appear to lie on a continuum in different hosts. Here, we consider how iron metabolism may represent a fundamental aspect of host homeostasis that is impacted by Wolbachia infection, connecting disparate pathways ranging from the provision of haem and ATP to programmed cell death, aging, and the recycling of intracellular resources. Depending on how Wolbachia and host cells interact across networks that depend on iron, the gradient between parasitism and mutualism may shift dynamically in some systems, or alternatively, stabilise on one or the other end of the spectrum.}, } @article {pmid25313210, year = {2015}, author = {Lentz, CS and Sattler, JM and Fendler, M and Gottwalt, S and Halls, VS and Strassel, S and Arriens, S and Hannam, JS and Specht, S and Famulok, M and Mueller, AK and Hoerauf, A and Pfarr, KM}, title = {In vitro activity of wALADin benzimidazoles against different life cycle stages of Plasmodium parasites.}, journal = {Antimicrobial agents and chemotherapy}, volume = {59}, number = {1}, pages = {654-658}, pmid = {25313210}, issn = {1098-6596}, mesh = {Antimalarials/*pharmacology ; Benzimidazoles/chemistry/*pharmacology ; Humans ; Inhibitory Concentration 50 ; Plasmodium berghei/drug effects ; Plasmodium falciparum/*drug effects/physiology ; Porphobilinogen Synthase/*antagonists & inhibitors ; Thiophenes/chemistry/pharmacology ; Toxoplasma/drug effects ; }, abstract = {wALADin1 benzimidazoles are specific inhibitors of δ-aminolevulinic acid dehydratase from Wolbachia endobacteria of filarial nematodes. We report that wALADin1 and two derivatives killed blood stage Plasmodium falciparum in vitro (50% inhibitory concentrations, 39, 7.7, and 12.8 μM, respectively). One of these derivatives inhibited gliding motility of Plasmodium berghei ANKA infectious sporozoites with nanomolar affinity and blocked invasion into hepatocytes but did not affect intrahepatocytic replication. Hence, wALADin1 benzimidazoles are tools to study gliding motility and potential antiplasmodial drug candidates.}, } @article {pmid25311369, year = {2014}, author = {Wang, GH and Niu, LM and Ma, GC and Xiao, JH and Huang, DW}, title = {Large proportion of genes in one cryptic WO prophage genome are actively and sex-specifically transcribed in a fig wasp species.}, journal = {BMC genomics}, volume = {15}, number = {1}, pages = {893}, pmid = {25311369}, issn = {1471-2164}, mesh = {Animals ; Female ; Ficus/parasitology ; Gene Expression Profiling ; *Gene Expression Regulation, Viral ; Genes, Viral ; Male ; Open Reading Frames ; Prophages/*genetics/physiology ; Sex Factors ; Wasps/*classification/genetics/*microbiology ; Wolbachia/genetics/physiology/*virology ; }, abstract = {BACKGROUND: Cryptic prophages are genetically defective in their induction and propagation, and are simply regarded as genetic remnants. There are several putative cryptic WO prophages in the sequenced Wolbachia genomes. Whether they are lytic is unclear and their functions are poorly understood. Only three open reading frames (ORFs) in cryptic WO prophages have been reported to be actively transcribed.

RESULTS: In this study, we comprehensively examined the transcription of the only cryptic WO prophage (WOSol) in a Wolbachia strain that infects a fig wasp, Ceratosolen solmsi (Agaonidae, Chalcidoidea). By analyzing the transcriptions of all the ORFs of WOSol in both sexes of C. solmsi, using qualitative and quantitative methods, we demonstrated that i) a high percentage of ORFs are actively transcribed (59%, 17/29); ii) the expression of these ORFs is highly sex-specific, with a strong male bias (three in females and 15 in males); iii) an ank (ankyrin-domain-containing) gene actively transcribed in both wasp sexes is more highly expressed in males.

CONCLUSIONS: A large proportion of the genes in the cryptic WO prophage WOSol are expressed, which overturns the concept that cryptic prophages are simply genetically defective. The highly sex-specific expression patterns of these genes in the host suggest that they play important roles in Wolbachia biology and its reproductive manipulation of its insect host, particularly through the males.}, } @article {pmid25300665, year = {2014}, author = {Fallon, AM}, title = {Flow cytometric evaluation of the intracellular bacterium, Wolbachia pipientis, in mosquito cells.}, journal = {Journal of microbiological methods}, volume = {107}, number = {}, pages = {119-125}, pmid = {25300665}, issn = {1872-8359}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI 081322/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Bacteriological Techniques ; Cell Line ; Culicidae/microbiology ; Cytoplasm/*microbiology ; Escherichia coli ; *Flow Cytometry/methods ; *Wolbachia/drug effects/physiology ; }, abstract = {Conditions for flow cytometric evaluation of the intracellular bacterium, Wolbachia pipientis, in infected mosquito cells are described. This approach will streamline investigation of Wolbachia's interactions with host cells and facilitate identification of culture conditions that select for Wolbachia-infected cells.}, } @article {pmid25297658, year = {2015}, author = {Kapantaidaki, DE and Ovčarenko, I and Fytrou, N and Knott, KE and Bourtzis, K and Tsagkarakou, A}, title = {Low levels of mitochondrial DNA and symbiont diversity in the worldwide agricultural pest, the greenhouse whitefly Trialeurodes vaporariorum (Hemiptera: Aleyrodidae).}, journal = {The Journal of heredity}, volume = {106}, number = {1}, pages = {80-92}, doi = {10.1093/jhered/esu061}, pmid = {25297658}, issn = {1465-7333}, mesh = {*Animal Distribution ; Animals ; Bacteria/*genetics ; Base Sequence ; DNA, Mitochondrial/*genetics ; Female ; *Genetic Variation ; Hemiptera/*genetics/microbiology ; Likelihood Functions ; Male ; Models, Genetic ; Molecular Sequence Data ; *Phylogeny ; Phylogeography ; Sequence Analysis, DNA ; Species Specificity ; Symbiosis/*genetics ; }, abstract = {Trialeurodes vaporariorum, the greenhouse whitefly, is a cosmopolitan agricultural pest. Little is known about the genetic diversity of T. vaporariorum and the bacterial symbionts associated with this species. Here, we undertook a large phylogeographic study by investigating both the mitochondrial (mt) diversity and the infection status of 38 T. vaporariorum collections from 18 countries around the world. Genetic diversity of T. vaporariorum was studied by analyzing sequence data from the mt cytochrome oxidase I, cytochrome b, and NADH dehydrogenase subunit 5 genes. Maximum-likelihood (ML) phylogeny reconstruction delineated 2 clades characterized by limited sequence divergence: one clade comprised samples only from the Northern hemisphere whereas the other comprised samples from a broader geographical range. The presence of secondary symbionts was determined by PCR using primers specific for Hamiltonella, Rickettsia, Arsenophonus, Cardinium, Wolbachia, and Fritschea. Most individuals examined harbored at least one secondary endosymbiont, and Arsenophonus was detected in almost all male and female individuals. Wolbachia was present at a much lower frequency, and Cardinium was detected in only a few individuals from Greece. Rickettsia, Hamiltonella, and Fritschea were not found. Additionally, we set out to further analyze Arsenophonus diversity by multilocus sequence typing analysis; however, the Arsenophonus sequences did not exhibit any polymorphism. Our results revealed remarkably low diversity in both mtDNA and symbionts in this worldwide agricultural pest, contrasting sharply with that of the ecologically similar Bemisia tabaci.}, } @article {pmid25287420, year = {2014}, author = {Voronin, D and Guimarães, AF and Molyneux, GR and Johnston, KL and Ford, L and Taylor, MJ}, title = {Wolbachia lipoproteins: abundance, localisation and serology of Wolbachia peptidoglycan associated lipoprotein and the Type IV Secretion System component, VirB6 from Brugia malayi and Aedes albopictus.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {462}, pmid = {25287420}, issn = {1756-3305}, mesh = {Aedes/*chemistry ; Animals ; Antibodies, Bacterial ; Bacterial Outer Membrane Proteins/chemistry/metabolism ; Bacterial Secretion Systems/*physiology ; Brugia malayi/*chemistry ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin G ; Lipoproteins/chemistry/*metabolism ; Peptidoglycan/chemistry/*metabolism ; Wolbachia/chemistry/*metabolism ; }, abstract = {BACKGROUND: Lipoproteins are the major agonists of Wolbachia-dependent inflammatory pathogenesis in filariasis and a validated target for drug discovery. Here we characterise the abundance, localisation and serology of the Wolbachia lipoproteins: Wolbachia peptidoglycan associated lipoprotein and the Type IV Secretion System component, VirB6.

METHODS: We used proteomics to confirm lipoprotein presence and relative abundance; fractionation, immunoblotting and confocal and electron immuno-microscopy for localisation and ELISA for serological analysis.

RESULTS: Proteomic analysis of Brugia malayi adult female protein extracts confirmed the presence of two lipoproteins, previously predicted through bioinformatics: Wolbachia peptidoglycan associated lipoprotein (wBmPAL) and the Type IV Secretion System component, VirB6 (wBmVirB6). wBmPAL was among the most abundant Wolbachia proteins present in an extract of adult female worms with wBmVirB6 only detected at a much lower abundance. This differential abundance was reflected in the immunogold-labelling, which showed wBmPAL localised at numerous sites within the bacterial membranes, whereas wBmVirB6 was present as a single cluster on each bacterial cell and also located within the bacterial membranes. Immunoblotting of fractionated extracts confirmed the localisation of wBmPAL to membranes and its absence from cytosolic fractions of C6/36 mosquito cells infected with wAlbB. In whole worm mounts, antibody labelling of both lipoproteins were associated with Wolbachia. Serological analysis showed that both proteins were immunogenic and raised antibody responses in the majority of individuals infected with Wuchereria bancrofti.

CONCLUSIONS: Two Wolbachia lipoproteins, wBmPAL and wBmVirB6, are present in extracts of Brugia malayi with wBmPAL among the most abundant of Wolbachia proteins. Both lipoproteins localised to bacterial membranes with wBmVirB6 present as a single cluster suggesting a single Type IV Secretory System on each Wolbachia cell.}, } @article {pmid25286971, year = {2014}, author = {Paulson, AR and von Aderkas, P and Perlman, SJ}, title = {Bacterial associates of seed-parasitic wasps (Hymenoptera: Megastigmus).}, journal = {BMC microbiology}, volume = {14}, number = {}, pages = {224}, pmid = {25286971}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/*genetics ; *Biota ; DNA, Bacterial/chemistry/genetics ; Female ; Hymenoptera/*microbiology ; Male ; Molecular Sequence Data ; Pseudotsuga/parasitology ; Seeds/parasitology ; Sequence Analysis, DNA ; }, abstract = {BACKGROUND: The success of herbivorous insects has been shaped largely by their association with microbes. Seed parasitism is an insect feeding strategy involving intimate contact and manipulation of a plant host. Little is known about the microbial associates of seed-parasitic insects. We characterized the bacterial symbionts of Megastigmus (Hymenoptera: Torymidae), a lineage of seed-parasitic chalcid wasps, with the goal of identifying microbes that might play an important role in aiding development within seeds, including supplementing insect nutrition or manipulating host trees. We screened multiple populations of seven species for common facultative inherited symbionts. We also performed culture independent surveys of larvae, pupae, and adults of M. spermotrophus using 454 pyrosequencing. This major pest of Douglas-fir is the best-studied Megastigmus, and was previously shown to manipulate its tree host into redirecting resources towards unfertilized ovules. Douglas-fir ovules and the parasitoid Eurytoma sp. were also surveyed using pyrosequencing to help elucidate possible transmission mechanisms of the microbial associates of M. spermotrophus.

RESULTS: Three wasp species harboured Rickettsia; two of these also harboured Wolbachia. Males and females were infected at similar frequencies, suggesting that these bacteria do not distort sex ratios. The M. spermotrophus microbiome is dominated by five bacterial OTUs, including lineages commonly found in other insect microbiomes and in environmental samples. The bacterial community associated with M. spermotrophus remained constant throughout wasp development and was dominated by a single OTU - a strain of Ralstonia, in the Betaproteobacteria, comprising over 55% of all bacterial OTUs from Megastigmus samples. This strain was also present in unparasitized ovules.

CONCLUSIONS: This is the first report of Ralstonia being an abundant and potentially important member of an insect microbiome, although other closely-related Betaproteobacteria, such as Burkholderia, are important insect symbionts. We speculate that Ralstonia might play a role in nutrient recycling, perhaps by redirecting nitrogen. The developing wasp larva feeds on megagametophyte tissue, which contains the seed storage reserves and is especially rich in nitrogen. Future studies using Ralstonia-specific markers will determine its distribution in other Megastigmus species, its mode of transmission, and its role in wasp nutrition.}, } @article {pmid25283608, year = {2014}, author = {Gerth, M and Gansauge, MT and Weigert, A and Bleidorn, C}, title = {Phylogenomic analyses uncover origin and spread of the Wolbachia pandemic.}, journal = {Nature communications}, volume = {5}, number = {}, pages = {5117}, doi = {10.1038/ncomms6117}, pmid = {25283608}, issn = {2041-1723}, mesh = {Algorithms ; Animals ; Arthropods ; Bacterial Proteins/genetics ; Cluster Analysis ; Evolution, Molecular ; *Genome, Bacterial ; Genomics ; Likelihood Functions ; *Phylogeny ; Reproducibility of Results ; Symbiosis/genetics ; Wolbachia/*genetics/metabolism ; }, abstract = {Of all obligate intracellular bacteria, Wolbachia is probably the most common. In general, Wolbachia are either widespread, opportunistic reproductive parasites of arthropods or essential mutualists in a single group of filarial nematodes, including many species of medical significance. To date, a robust phylogenetic backbone of Wolbachia is lacking and consequently, many Wolbachia-related phenomena cannot be discussed in a broader evolutionary context. Here we present the first comprehensive phylogenomic analysis of Wolbachia supergroup relationships based on new whole-genome-shotgun data. Our results suggest that Wolbachia has switched between its two major host groups at least twice. The ability of some arthropod-infecting Wolbachia to universally infect and to adapt to a broad range of hosts quickly is restricted to a single monophyletic lineage (containing supergroups A and B). Thus, the currently observable pandemic has likely a single evolutionary origin and is unique within the radiation of Wolbachia strains.}, } @article {pmid25278497, year = {2015}, author = {Clare, RH and Cook, DA and Johnston, KL and Ford, L and Ward, SA and Taylor, MJ}, title = {Development and validation of a high-throughput anti-Wolbachia whole-cell screen: a route to macrofilaricidal drugs against onchocerciasis and lymphatic filariasis.}, journal = {Journal of biomolecular screening}, volume = {20}, number = {1}, pages = {64-69}, doi = {10.1177/1087057114551518}, pmid = {25278497}, issn = {1552-454X}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; Antinematodal Agents/*pharmacology/therapeutic use ; Cell Line ; Dose-Response Relationship, Drug ; Drug Discovery/*methods/standards ; Elephantiasis, Filarial/drug therapy ; High-Throughput Screening Assays/*methods/standards ; Humans ; Neglected Diseases/drug therapy ; Onchocerca/microbiology ; Onchocerciasis/drug therapy ; Reproducibility of Results ; Wolbachia/*drug effects/*physiology ; }, abstract = {There is an urgent need to develop new, safe, and affordable macrofilaricidal drugs for onchocerciasis and lymphatic filariasis treatment and control. The Anti-Wolbachia Consortium (A·WOL) aims to provide a novel treatment with macrofilaricidal activity by targeting the essential bacterial symbiont Wolbachia. The consortium is currently screening a diverse range of compounds to find new chemical space to drive this drug discovery initiative and address this unmet demand. To increase the throughput and capacity of the A·WOL cell-based screen, we have developed a 384-well format assay using a high-content imaging system (Operetta) in conjunction with optimized Wolbachia growth dynamics in the C6/36 Aedes albopictus mosquito cell line. This assay uses texture analysis of cells stained with SYTO 11 as a direct measure of bacterial load. This validated assay has dramatically increased the capacity and throughput of the A·WOL compound library screening program 25-fold, enriching the number of new anti-Wolbachia hits identified for further development as potential macrofilaricides for onchocerciasis and lymphatic filariasis.}, } @article {pmid25266919, year = {2014}, author = {Oteo, JA and Portillo, A and Portero, F and Zavala-Castro, J and Venzal, JM and Labruna, MB}, title = {'Candidatus Rickettsia asemboensis' and Wolbachia spp. in Ctenocephalides felis and Pulex irritans fleas removed from dogs in Ecuador.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {455}, pmid = {25266919}, issn = {1756-3305}, mesh = {Animals ; Base Sequence ; Ctenocephalides/microbiology ; DNA, Bacterial/chemistry ; Dog Diseases/*microbiology/parasitology/transmission ; Dogs ; Ecuador ; Female ; Flea Infestations/parasitology/*veterinary ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/*isolation & purification ; Rickettsia Infections/microbiology/transmission ; Rickettsiaceae Infections/*microbiology/transmission ; Sequence Analysis, DNA ; Siphonaptera/*microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Flea-borne infections are distributed worldwide. Up to date there are no reports about microorganisms associated to fleas in Ecuador.

METHODS: Seventy-one Pulex irritans and 8 Ctenocephalides felis fleas were removed from dogs in two Ecuadorian areas (Pastaza and Chimborazo Provinces) in December 2012. DNA extracts were tested by polymerase chain reaction (PCR) assays targeting universal 16S rRNA, as well as screened for the presence of Rickettsia spp. (gltA, htrA, ompB, sca4 and ompA genes) and Bartonella spp. (rpoB, gltA and ITS genes).

RESULTS: Our results showed the presence of 'Candidatus Rickettsia asemboensis' (highly similar to R. felis) in C. felis and Wolbachia spp. endosimbionts in P. irritans collected from animals in Ecuador. No fleas were found to be positive for any Bartonella species or Yersinia pestis.

CONCLUSIONS: Clinicians should be aware of the potential risk of this new Candidatus Rickettsia sp. and keep in mind other flea-borne infections since these flea species frequently bite humans.}, } @article {pmid25259930, year = {2014}, author = {Marubayashi, JM and Kliot, A and Yuki, VA and Rezende, JA and Krause-Sakate, R and Pavan, MA and Ghanim, M}, title = {Diversity and localization of bacterial endosymbionts from whitefly species collected in Brazil.}, journal = {PloS one}, volume = {9}, number = {9}, pages = {e108363}, pmid = {25259930}, issn = {1932-6203}, mesh = {Animals ; Brazil ; Enterobacteriaceae/*genetics ; Hemiptera/*microbiology ; Molecular Sequence Data ; Symbiosis/*genetics ; }, abstract = {Whiteflies (Hemiptera: Aleyrodidae) are sap-sucking insect pests, and some cause serious damage in agricultural crops by direct feeding and by transmitting plant viruses. Whiteflies maintain close associations with bacterial endosymbionts that can significantly influence their biology. All whitefly species harbor a primary endosymbiont, and a diverse array of secondary endosymbionts. In this study, we surveyed 34 whitefly populations collected from the states of Sao Paulo, Bahia, Minas Gerais and Parana in Brazil, for species identification and for infection with secondary endosymbionts. Sequencing the mitochondrial Cytochrome Oxidase I gene revealed the existence of five whitefly species: The sweetpotato whitefly Bemisia tabaci B biotype (recently termed Middle East-Asia Minor 1 or MEAM1), the greenhouse whitefly Trialeurodes vaporariorum, B. tabaci A biotype (recently termed New World 2 or NW2) collected only from Euphorbia, the Acacia whitefly Tetraleurodes acaciae and Bemisia tuberculata both were detected only on cassava. Sequencing rRNA genes showed that Hamiltonella and Rickettsia were highly prevalent in all MEAM1 populations, while Cardinium was close to fixation in only three populations. Surprisingly, some MEAM1 individuals and one NW2 population were infected with Fritschea. Arsenopnohus was the only endosymbiont detected in T. vaporariorum. In T. acaciae and B. tuberculata populations collected from cassava, Wolbachia was fixed in B. tuberculata and was highly prevalent in T. acaciae. Interestingly, while B. tuberculata was additionally infected with Arsenophonus, T. acaciae was infected with Cardinium and Fritschea. Fluorescence in situ hybridization analysis on representative individuals showed that Hamiltonella, Arsenopnohus and Fritschea were localized inside the bacteriome, Cardinium and Wolbachia exhibited dual localization patterns inside and outside the bacteriome, and Rickettsia showed strict localization outside the bacteriome. This study is the first survey of whitely populations collected in Brazil, and provides further insights into the complexity of infection with secondary endosymionts in whiteflies.}, } @article {pmid25259690, year = {2014}, author = {Hoffmann, M and Coy, MR and Kingdom Gibbard, HN and Pelz-Stelinski, KS}, title = {Wolbachia infection density in populations of the Asian citrus psyllid (Hemiptera: Liviidae).}, journal = {Environmental entomology}, volume = {43}, number = {5}, pages = {1215-1222}, doi = {10.1603/EN14193}, pmid = {25259690}, issn = {1938-2936}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics/metabolism ; Base Sequence ; Citrus/growth & development ; Female ; Florida ; Hemiptera/*microbiology ; Male ; Molecular Sequence Data ; Real-Time Polymerase Chain Reaction ; Symbiosis ; Wolbachia/*genetics/isolation & purification/metabolism ; }, abstract = {The symbiotic relationships between bacteria of the genus Wolbachia (order Rickettsiales) and their arthropod hosts are diverse and can range from mutualism to parasitism. Whereas effects of Wolbachia on host biology are well investigated, little is known about diversity and abundance of Wolbachia in their natural hosts. The phloem-feeding Asian citrus psyllid, Diaphorina citri (Kuwayama) (Hemiptera: Liviidae), is naturally infected with Wolbachia (wDi). In the current study, we calculated the within-host density of Wolbachia in Florida D. citri populations using quantitative polymerase chain reaction for detection of the Wolbachia outer surface protein gene, wsp. Gene quantities were normalized to the D. citri wingless gene (Wg) to estimate Wolbachia abundance in individual D. citri. Using this method, significant geographic differences in Wolbachia densities were detected among Florida D. citri populations, with higher infection levels occurring in male versus female hosts.}, } @article {pmid25253218, year = {2014}, author = {Sivan, A and Shriram, AN and Bhattacharya, D and Vijayachari, P}, title = {Wolbachia endobacterium in wild population of Aedes albopictus (Skuse) (Diptera: Culicidae) and phylogeny from Andaman and Nicobar Islands, India.}, journal = {Journal of vector borne diseases}, volume = {51}, number = {3}, pages = {235-238}, pmid = {25253218}, issn = {0972-9062}, mesh = {Aedes/*microbiology ; Animals ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; India ; *Phylogeny ; Sequence Analysis, DNA ; Wolbachia/*classification/*genetics/isolation & purification ; }, } @article {pmid25250237, year = {2014}, author = {Mahalingashetti, PB and Subramanian, RA and Jayker, SS and Vijay, A}, title = {Lymphatic filariasis: A view at pathological diversity.}, journal = {Tropical parasitology}, volume = {4}, number = {2}, pages = {128-132}, pmid = {25250237}, issn = {2229-5070}, abstract = {Filariasis is traditionally diagnosed following screening of peripheral smear for microfilaria. Clinically lymphatic filariasis mimics the common local diseases. Thus, it is plausible to observe this parasitic infection in histological sections. We encountered three such cases, which displayed diverse patterns of immune response. Presence of both dead and viable worm at the same foci suggests that such immune response could be the result of parasitic death. Histological features such as endothelial injury and granulomatous response attests to the role of Wolbachia bacteria in influencing tissue response.}, } @article {pmid25249781, year = {2014}, author = {Zha, X and Zhang, W and Zhou, C and Zhang, L and Xiang, Z and Xia, Q}, title = {Detection and characterization of Wolbachia infection in silkworm.}, journal = {Genetics and molecular biology}, volume = {37}, number = {3}, pages = {573-580}, pmid = {25249781}, issn = {1415-4757}, support = {Z01 AA000306//Intramural NIH HHS/United States ; }, abstract = {Wolbachia naturally infects a wide variety of arthropods, where it plays important roles in host reproduction. It was previously reported that Wolbachia did not infect silkworm. By means of PCR and sequencing we found in this study that Wolbachia is indeed present in silkworm. Phylogenetic analysis indicates that Wolbachia infection in silkworm may have occurred via transfer from parasitic wasps. Furthermore, Southern blotting results suggest a lateral transfer of the wsp gene into the genomes of some wild silkworms. By antibiotic treatments, we found that tetracycline and ciprofloxacin can eliminate Wolbachia in the silkworm and Wolbachia is important to ovary development of silkworm. These results provide clues towards a more comprehensive understanding of the interaction between Wolbachia and silkworm and possibly other lepidopteran insects.}, } @article {pmid25249342, year = {2014}, author = {Martínez-Rodríguez, P and Granero-Belinchón, R and Arroyo-Yebras, F and Bella, JL}, title = {New insight into Wolbachia epidemiology: its varying incidence during the host life cycle can alter bacteria spread.}, journal = {Bulletin of mathematical biology}, volume = {76}, number = {10}, pages = {2646-2663}, doi = {10.1007/s11538-014-0029-5}, pmid = {25249342}, issn = {1522-9602}, mesh = {Animals ; Biological Evolution ; Computer Simulation ; Female ; Grasshoppers/growth & development/microbiology/physiology ; Host-Pathogen Interactions ; Life Cycle Stages ; Male ; Mathematical Concepts ; Models, Biological ; Reproduction ; Symbiosis ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Wolbachia is an obligate endosymbiont whose spread depends mainly on its capacity to alter host reproduction by, for instance, cytoplasmic incompatibility. Several mathematical models have been developed to explain the dynamics of bacterial spread, because of its applied interest. However, some aspects of the host's and bacterium's biology have not been considered in modelling: for instance, changes in Wolbachia proportions during the host's life cycle have been observed in several species, including Drosophila sp., Nasonia sp. and Aedes sp. (Diptera), but also in the grasshopper Chorthippus parallelus (Orthoptera), the species studied in this article. These changes influence the proportion of incompatible crosses and, consequently, infection prevalence in subsequent generations. In this paper, we are interested in ascertaining whether these changes in the infection proportions during the host's life cycle can influence the dynamics of the spread of these bacteria. We have examined its consequences using a mathematical model to predict the evolution of Wolbachia infection frequencies. The simulations were validated by experimental field data from C. parallelus. The main outcome is that those changes above mentioned might affect long-term infection spread, with possible consequences for the current distribution of Wolbachia and the way it affects its host's reproduction.}, } @article {pmid25233341, year = {2014}, author = {Martinez, J and Longdon, B and Bauer, S and Chan, YS and Miller, WJ and Bourtzis, K and Teixeira, L and Jiggins, FM}, title = {Symbionts commonly provide broad spectrum resistance to viruses in insects: a comparative analysis of Wolbachia strains.}, journal = {PLoS pathogens}, volume = {10}, number = {9}, pages = {e1004369}, pmid = {25233341}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; P 22634/FWF_/Austrian Science Fund FWF/Austria ; 094664MA/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila/*growth & development/*immunology/microbiology/virology ; Female ; Host-Pathogen Interactions/*immunology ; Insect Viruses/*pathogenicity ; Male ; Real-Time Polymerase Chain Reaction ; Symbiosis/*immunology ; Virus Diseases/*immunology/microbiology/virology ; Wolbachia/classification/*physiology ; }, abstract = {In the last decade, bacterial symbionts have been shown to play an important role in protecting hosts against pathogens. Wolbachia, a widespread symbiont in arthropods, can protect Drosophila and mosquito species against viral infections. We have investigated antiviral protection in 19 Wolbachia strains originating from 16 Drosophila species after transfer into the same genotype of Drosophila simulans. We found that approximately half of the strains protected against two RNA viruses. Given that 40% of terrestrial arthropod species are estimated to harbour Wolbachia, as many as a fifth of all arthropods species may benefit from Wolbachia-mediated protection. The level of protection against two distantly related RNA viruses--DCV and FHV--was strongly genetically correlated, which suggests that there is a single mechanism of protection with broad specificity. Furthermore, Wolbachia is making flies resistant to viruses, as increases in survival can be largely explained by reductions in viral titer. Variation in the level of antiviral protection provided by different Wolbachia strains is strongly genetically correlated to the density of the bacteria strains in host tissues. We found no support for two previously proposed mechanisms of Wolbachia-mediated protection--activation of the immune system and upregulation of the methyltransferase Dnmt2. The large variation in Wolbachia's antiviral properties highlights the need to carefully select Wolbachia strains introduced into mosquito populations to prevent the transmission of arboviruses.}, } @article {pmid25230723, year = {2014}, author = {Badawi, M and Giraud, I and Vavre, F and Grève, P and Cordaux, R}, title = {Signs of neutralization in a redundant gene involved in homologous recombination in Wolbachia endosymbionts.}, journal = {Genome biology and evolution}, volume = {6}, number = {10}, pages = {2654-2664}, pmid = {25230723}, issn = {1759-6653}, mesh = {Evolution, Molecular ; Genome, Bacterial/*genetics ; Phylogeny ; Symbiosis/genetics/physiology ; Wolbachia/*genetics ; }, abstract = {Genomic reduction in bacterial endosymbionts occurs through large genomic deletions and long-term accumulation of mutations. The latter process involves successive steps including gene neutralization, pseudogenization, and gradual erosion until complete loss. Although many examples of pseudogenes at various levels of degradation have been reported, neutralization cases are scarce because of the transient nature of the process. Gene neutralization may occur due to relaxation of selection in nonessential genes, for example, those involved in redundant functions. Here, we report an example of gene neutralization in the homologous recombination (HR) pathway of Wolbachia, a bacterial endosymbiont of arthropods and nematodes. The HR pathway is often depleted in endosymbiont genomes, but it is apparently intact in some Wolbachia strains. Analysis of 12 major HR genes showed that they have been globally under strong purifying selection during the evolution of Wolbachia strains hosted by arthropods, supporting the evolutionary importance of the HR pathway for these Wolbachia genomes. However, we detected signs of recent neutralization of the ruvA gene in a subset of Wolbachia strains, which might be related to an ancestral, clade-specific amino acid change that impaired DNA-binding activity. Strikingly, RuvA is part of the RuvAB complex involved in branch migration, whose function overlaps with the RecG helicase. Although ruvA is experiencing neutralization, recG is under strong purifying selection. Thus, our high phylogenetic resolution suggests that we identified a rare example of targeted neutralization of a gene involved in a redundant function in an endosymbiont genome.}, } @article {pmid25228147, year = {2014}, author = {Zélé, F and Vézilier, J and L'Ambert, G and Nicot, A and Gandon, S and Rivero, A and Duron, O}, title = {Dynamics of prevalence and diversity of avian malaria infections in wild Culex pipiens mosquitoes: the effects of Wolbachia, filarial nematodes and insecticide resistance.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {437}, pmid = {25228147}, issn = {1756-3305}, support = {243054/ERC_/European Research Council/International ; }, mesh = {Animals ; Birds ; Coinfection/veterinary ; Culex/*parasitology ; Disease Reservoirs ; Female ; France/epidemiology ; Haemosporida/physiology ; Insect Vectors/*parasitology ; *Insecticide Resistance ; Malaria, Avian/*epidemiology/parasitology/transmission ; Models, Theoretical ; Nematoda/physiology ; Phylogeny ; Plasmodium/*physiology ; Prevalence ; Seasons ; Wolbachia/physiology ; }, abstract = {BACKGROUND: Identifying the parasites transmitted by a particular vector and the factors that render this vector susceptible to the parasite are key steps to understanding disease transmission. Although avian malaria has become a model system for the investigation of the ecological and evolutionary dynamics of Plasmodium parasites, little is still known about the field prevalence, diversity and distribution of avian Plasmodium species within the vectors, or about the extrinsic factors affecting Plasmodium population dynamics in the wild.

METHODS: We examined changes in avian malaria prevalence and Plasmodium lineage composition in female Culex pipiens caught throughout one field season in 2006, across four sampling sites in southern France. Using site occupancy models, we correct the naive estimates of Plasmodium prevalence to account for PCR-based imperfect detection. To establish the importance of different factors that may bear on the prevalence and diversity of avian Plasmodium in field mosquitoes, we focus on Wolbachia and filarial parasite co-infections, as well as on the insecticide resistance status of the mosquito.

RESULTS: Plasmodium prevalence in Cx. pipiens increased from February (0%) to October (15.8%) and did not vary significantly among the four sampling sites. The application of site occupancy models leads to a 4% increase in this initial (naive) estimate of prevalence. The parasite community was composed of 15 different haemosporidian lineages, 13 of which belonged to the Plasmodium genus, and 2 to the Haemoproteus genus. Neither the presence of different Wolbachia types and of filarial parasites co-infecting the mosquitoes, nor their insecticide resistance status were found to affect the Plasmodium prevalence and diversity.

CONCLUSION: We found that haemosporidian parasites are common and diverse in wild-caught Cx. pipiens mosquitoes in Southern France. The prevalence of the infection in mosquitoes is unaffected by Wolbachia and filarial co-infections as well as the insecticide resistant status of the vector. These factors may thus have a negligible impact on the transmission of avian malaria. In contrast, the steady increase in prevalence from February to October indicates that the dynamics of avian malaria is driven by seasonality and supports that infected birds are the reservoir of a diverse community of lineages in southern France.}, } @article {pmid25225494, year = {2014}, author = {Asgharian, H and Chang, PL and Mazzoglio, PJ and Negri, I}, title = {Wolbachia is not all about sex: male-feminizing Wolbachia alters the leafhopper Zyginidia pullula transcriptome in a mainly sex-independent manner.}, journal = {Frontiers in microbiology}, volume = {5}, number = {}, pages = {430}, pmid = {25225494}, issn = {1664-302X}, abstract = {Wolbachia causes the feminization of chromosomally male embryos in several species of crustaceans and insects, including the leafhopper Zyginidia pullula. In contrast to the relatively well-established ecological aspects of male feminization (e.g., sex ratio distortion and its consequences), the underlying molecular mechanisms remain understudied and unclear. We embarked on an exploratory study to investigate the extent and nature of Wolbachia's effect on gene expression pattern in Z. pullula. We sequenced whole transcriptomes from Wolbachia-infected and uninfected adults. 18147 loci were assembled de novo, including homologs of several Drosophila sex determination genes. A number of transcripts were flagged as candidate Wolbachia sequences. Despite the resemblance of Wolbachia-infected chromosomal males to uninfected and infected chromosomal females in terms of sexual morphology and behavior, principal component analysis revealed that gene expression patterns did not follow these sexual phenotype categories. The principal components generated by differentially expressed genes specified a strong sex-independent Wolbachia effect, followed by a weaker Wolbachia-sexual karyotype interaction effect. Approaches to further examine the molecular mechanism of Wolbachia-host interactions have been suggested based on the presented findings.}, } @article {pmid25224730, year = {2015}, author = {Zhang, YK and Ding, XL and Rong, X and Hong, XY}, title = {How do hosts react to endosymbionts? A new insight into the molecular mechanisms underlying the Wolbachia-host association.}, journal = {Insect molecular biology}, volume = {24}, number = {1}, pages = {1-12}, doi = {10.1111/imb.12128}, pmid = {25224730}, issn = {1365-2583}, mesh = {Animals ; Cytoplasm/physiology ; Female ; Gene Expression Profiling ; Male ; Oxidation-Reduction ; Phenotype ; Real-Time Polymerase Chain Reaction ; Reproduction/genetics ; Symbiosis/genetics ; Tetranychidae/*genetics/immunology/*microbiology ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {Wolbachia is an intracellular bacterium that has aroused intense interest because of its ability to alter the biology of its host in diverse ways. In the two-spotted spider mite, Tetranychus urticae, Wolbachia can induce complex cytoplasmic incompatibility (CI) phenotypes and fitness changes, although little is known about the mechanisms. In the present study, we selected a strain of T. urticae, in which Wolbachia infection was associated with strong CI and enhanced female fecundity, to investigate changes in the transcriptome of T. urticae in Wolbachia-infected vs. uninfected lines. The responses were found to be sex-specific, with the transcription of 251 genes being affected in females and 171 genes being affected in males. Some of the more profoundly affected genes in both sexes were lipocalin genes and genes involved in oxidation reduction, digestion and detoxification. Several of the differentially expressed genes have potential roles in reproduction. Interestingly, unlike certain Wolbachia transinfections in novel hosts, the Wolbachia-host association in the present study showed no clear evidence of host immune priming by Wolbachia, although a few potential immune genes were affected.}, } @article {pmid25211492, year = {2014}, author = {Hoffmann, AA and Iturbe-Ormaetxe, I and Callahan, AG and Phillips, BL and Billington, K and Axford, JK and Montgomery, B and Turley, AP and O'Neill, SL}, title = {Stability of the wMel Wolbachia Infection following invasion into Aedes aegypti populations.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {9}, pages = {e3115}, pmid = {25211492}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Female ; Host-Pathogen Interactions ; Humans ; Queensland ; Wolbachia/*physiology ; }, abstract = {The wMel infection of Drosophila melanogaster was successfully transferred into Aedes aegypti mosquitoes where it has the potential to suppress dengue and other arboviruses. The infection was subsequently spread into two natural populations at Yorkeys Knob and Gordonvale near Cairns, Queensland in 2011. Here we report on the stability of the infection following introduction and we characterize factors influencing the ongoing dynamics of the infection in these two populations. While the Wolbachia infection always remained high and near fixation in both locations, there was a persistent low frequency of uninfected mosquitoes. These uninfected mosquitoes showed weak spatial structure at both release sites although there was some clustering around two areas in Gordonvale. Infected females from both locations showed perfect maternal transmission consistent with patterns previously established pre-release in laboratory tests. After >2 years under field conditions, the infection continued to show complete cytoplasmic incompatibility across multiple gonotrophic cycles but persistent deleterious fitness effects, suggesting that host effects were stable over time. These results point to the stability of Wolbachia infections and their impact on hosts following local invasion, and also highlight the continued persistence of uninfected individuals at a low frequency most likely due to immigration.}, } @article {pmid25187984, year = {2014}, author = {Hughes, GL and Rivero, A and Rasgon, JL}, title = {Wolbachia can enhance Plasmodium infection in mosquitoes: implications for malaria control?.}, journal = {PLoS pathogens}, volume = {10}, number = {9}, pages = {e1004182}, pmid = {25187984}, issn = {1553-7374}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culicidae/*microbiology/*parasitology ; Host-Parasite Interactions ; Humans ; Malaria, Falciparum/parasitology/*prevention & control ; *Pest Control, Biological ; Plasmodium falciparum/*growth & development ; Wolbachia/*physiology ; }, } @article {pmid25184624, year = {2014}, author = {Rogers, EE and Backus, EA}, title = {Anterior foregut microbiota of the glassy-winged sharpshooter explored using deep 16S rRNA gene sequencing from individual insects.}, journal = {PloS one}, volume = {9}, number = {9}, pages = {e106215}, pmid = {25184624}, issn = {1932-6203}, mesh = {Animals ; Bacterial Proteins/genetics ; Base Sequence ; Cytoskeletal Proteins/genetics ; Digestive System/*microbiology ; Hemiptera/*microbiology ; High-Throughput Nucleotide Sequencing ; Host-Pathogen Interactions ; Insect Vectors/*microbiology ; Introduced Species ; Microbiota/*genetics ; Molecular Sequence Data ; Phylogeny ; Plant Diseases/microbiology ; RNA, Ribosomal, 16S/chemistry/*genetics ; Vitis/microbiology ; Wolbachia/classification/genetics ; Xylella/classification/*genetics ; }, abstract = {The glassy-winged sharpshooter (GWSS) is an invasive insect species that transmits Xylella fastidiosa, the bacterium causing Pierce's disease of grapevine and other leaf scorch diseases. X. fastidiosa has been shown to colonize the anterior foregut (cibarium and precibarium) of sharpshooters, where it may interact with other naturally-occurring bacterial species. To evaluate such interactions, a comprehensive list of bacterial species associated with the sharpshooter cibarium and precibarium is needed. Here, a survey of microbiota associated with the GWSS anterior foregut was conducted. Ninety-six individual GWSS, 24 from each of 4 locations (Bakersfield, CA; Ojai, CA; Quincy, FL; and a laboratory colony), were characterized for bacteria in dissected sharpshooter cibaria and precibaria by amplification and sequencing of a portion of the 16S rRNA gene using Illumina MiSeq technology. An average of approximately 150,000 sequence reads were obtained per insect. The most common genus detected was Wolbachia; sequencing of the Wolbachia ftsZ gene placed this strain in supergroup B, one of two Wolbachia supergroups most commonly associated with arthropods. X. fastidiosa was detected in all 96 individuals examined. By multilocus sequence typing, both X. fastidiosa subspecies fastidiosa and subspecies sandyi were present in GWSS from California and the colony; only subspecies fastidiosa was detected in GWSS from Florida. In addition to Wolbachia and X. fastidiosa, 23 other bacterial genera were detected at or above an average incidence of 0.1%; these included plant-associated microbes (Methylobacterium, Sphingomonas, Agrobacterium, and Ralstonia) and soil- or water-associated microbes (Anoxybacillus, Novosphingobium, Caulobacter, and Luteimonas). Sequences belonging to species of the family Enterobacteriaceae also were detected but it was not possible to assign these to individual genera. Many of these species likely interact with X. fastidiosa in the cibarium and precibarium.}, } @article {pmid25183276, year = {2014}, author = {Joanne, S and Vythilingam, I and Yugavathy, N and Doss, JI}, title = {Modified technique of Wolbachia removal from Malaysian Aedes albopictus.}, journal = {Asian Pacific journal of tropical biomedicine}, volume = {4}, number = {7}, pages = {557-560}, pmid = {25183276}, issn = {2221-1691}, abstract = {OBJECTIVE: To develop an artificial and modified Wolbachia removal technique using tetracycline from naturally Wolbachia infected Aedes albopictus (Ae. albopictus) so as to be able to produce generations of Wolbachia free offsprings.

METHODS: In this study, seven different tetracycline treatment methods were conducted to obtain the best removal method. Four methods focused on larvae tetracycline treatment, one method on both larvae and adult tetracycline treatment and the last two methods on adult mosquito sucrose treatment.

RESULTS: All larval tetracycline treatments resulted in either high larvae mortality, sterile F0 adult mosquitoes or unsuccessful Wolbachia removal. Treatment of both larvae and adults resulted in reduced larvae mortality, successful Wolbachia removal but slow mosquito fecundity. As for the adult treatment, 1.0 mg/mL as previously published was not able to completely remove Wolbachia in F1 generation whereas 1.25 mg/mL successfully removed Wolbachia from F1 and F2 mosquitoes in 2 weeks.

CONCLUSIONS: This method is different from the previously published methods as it provides an improved Wolbachia removal technique from Ae. albopictus with high egg hatchability, low larvae mortality, increased fecundity and better Wolbachia removal rate.}, } @article {pmid25182613, year = {2014}, author = {Montagna, M and Chouaia, B and Sacchi, L and Porretta, D and Martin, E and Giorgi, A and Lozzia, GC and Epis, S}, title = {A new strain of Wolbachia in an alpine population of the viviparous Oreina cacaliae (Coleoptera: Chrysomelidae).}, journal = {Environmental entomology}, volume = {43}, number = {4}, pages = {913-922}, doi = {10.1603/EN13228}, pmid = {25182613}, issn = {1938-2936}, mesh = {Animals ; Coleoptera/*microbiology/*physiology ; DNA, Bacterial/genetics/metabolism ; Female ; Male ; Multilocus Sequence Typing ; Oocytes/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics/metabolism ; Reproduction ; Sequence Analysis, DNA ; Wolbachia/classification/*genetics/physiology ; }, abstract = {Microbial symbionts played a central role in insect evolution. Oreina cacaliae (Schrank, 1785) (Coleoptera: Chrysomelidae) is a rare example of a viviparous insect, able to feed on toxic plants and sequester toxic compounds. In the current study, the microbiota associated with O. cacaliae was characterized using a culture-independent approach, targeting the 16S rRNA bacterial gene. The obtained 16S rRNA gene sequences were analyzed and identified at different taxonomic levels. Wolbachia was the dominant bacterium, both in male and female (100 and 91.9%, respectively) individuals; the detected Wolbachia was described as a new sequence type based on multilocus sequence typing (Wolbachia ST375 Ocac_A_wVdO). After phylogenetic analyses, Wolbachia ST375 Ocac_A_wVdO was attributed to the supergroup A. Immunofluorescence assays and electron microscopy confirmed the presence of Wolbachia within O. cacaliae oocytes, confirming its transovarial transmission in this species. Representatives of six species of Oreina were tested for the presence of Wolbachia through specific polymerase chain reaction, and a dendrogram was generated for these species based on coxI gene sequences. The Wolbachia harbored by different species of Oreina were characterized by multilocus sequence typing. Five out of the six examined Oreina species were positive for Wolbachia, with four of these harboring the same sequence type.}, } @article {pmid25182324, year = {2014}, author = {Touret, F and Guiguen, F and Terzian, C}, title = {Wolbachia influences the maternal transmission of the gypsy endogenous retrovirus in Drosophila melanogaster.}, journal = {mBio}, volume = {5}, number = {5}, pages = {e01529-14}, pmid = {25182324}, issn = {2150-7511}, mesh = {Animals ; DNA-Binding Proteins/genetics/metabolism ; Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster/genetics/*microbiology/*virology ; Endogenous Retroviruses/*physiology ; Female ; Male ; Oocytes/microbiology/virology ; Retroelements ; Symbiosis ; Transcription Factors/genetics/metabolism ; Viral Envelope Proteins/genetics/*metabolism ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {UNLABELLED: The endosymbiotic bacteria of the genus Wolbachia are present in most insects and are maternally transmitted through the germline. Moreover, these intracellular bacteria exert antiviral activity against insect RNA viruses, as in Drosophila melanogaster, which could explain the prevalence of Wolbachia bacteria in natural populations. Wolbachia is maternally transmitted in D. melanogaster through a mechanism that involves distribution at the posterior pole of mature oocytes and then incorporation into the pole cells of the embryos. In parallel, maternal transmission of several endogenous retroviruses is well documented in D. melanogaster. Notably, gypsy retrovirus is expressed in permissive follicle cells and transferred to the oocyte and then to the offspring by integrating into their genomes. Here, we show that the presence of Wolbachia wMel reduces the rate of gypsy insertion into the ovo gene. However, the presence of Wolbachia does not modify the expression levels of gypsy RNA and envelope glycoprotein from either permissive or restrictive ovaries. Moreover, Wolbachia affects the pattern of distribution of the retroviral particles and the gypsy envelope protein in permissive follicle cells. Altogether, our results enlarge the knowledge of the antiviral activity of Wolbachia to include reducing the maternal transmission of endogenous retroviruses in D. melanogaster.

IMPORTANCE: Animals have established complex relationships with bacteria and viruses that spread horizontally among individuals or are vertically transmitted, i.e., from parents to offspring. It is well established that members of the genus Wolbachia, maternally inherited symbiotic bacteria present mainly in arthropods, reduce the replication of several RNA viruses transmitted horizontally. Here, we demonstrate for the first time that Wolbachia diminishes the maternal transmission of gypsy, an endogenous retrovirus in Drosophila melanogaster. We hypothesize that gypsy cannot efficiently integrate into the germ cells of offspring during embryonic development in the presence of Wolbachia because both are competitors for localization to the posterior pole of the egg. More generally, it would be of interest to analyze the influence of Wolbachia on vertically transmitted exogenous viruses, such as some arboviruses.}, } @article {pmid25171611, year = {2014}, author = {Hoffmann, AA and Goundar, AA and Long, SA and Johnson, PH and Ritchie, SA}, title = {Invasion of Wolbachia at the residential block level is associated with local abundance of Stegomyia aegypti, yellow fever mosquito, populations and property attributes.}, journal = {Medical and veterinary entomology}, volume = {28 Suppl 1}, number = {}, pages = {90-97}, doi = {10.1111/mve.12077}, pmid = {25171611}, issn = {1365-2915}, mesh = {Aedes/*microbiology/physiology ; Animals ; Female ; Geography ; *Mosquito Control ; Population Density ; Queensland ; Residence Characteristics ; Wolbachia/*physiology ; }, abstract = {Wolbachia can suppress dengue and control mosquito populations and this depends on the successful invasion of Wolbachia-infected mosquitoes into local populations. Ovitrap data collected during the recent invasion of wMel-infected Stegomyia aegypti (Diptera: Culicidae) (Linnaeus) into Gordonvale near Cairns, Australia, were used to identify variables that help predict the success of localized invasion. Based on the variance in Wolbachia frequencies across Gordonvale as well as at another release site at Yorkeys Knob in comparison to simulations, it was estimated that on average 2-4 females contributed eggs to an ovitrap. By collating ovitrap data from two collection periods at the start of the release from residential blocks, it was found that uninfected mosquitoes had a patchy distribution across the release site. Residential blocks with relatively high uninfected mosquito numbers were less easily invaded by Wolbachia than blocks with low numbers. The numbers of uninfected mosquitoes in ovitraps were negatively correlated with the proportion of brick houses in a residential block, whereas local Wolbachia frequencies were correlated positively with this variable as well as negatively with the amount of shading in a yard and availability of breeding sites. These findings point to proxy measures for predicting the ease of localized invasion of Wolbachia.}, } @article {pmid25165813, year = {2014}, author = {Landmann, F and Foster, JM and Michalski, ML and Slatko, BE and Sullivan, W}, title = {Co-evolution between an endosymbiont and its nematode host: Wolbachia asymmetric posterior localization and AP polarity establishment.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {8}, pages = {e3096}, pmid = {25165813}, issn = {1935-2735}, mesh = {Animals ; *Biological Evolution ; Brugia malayi/genetics/*microbiology/physiology ; Cell Polarity/*physiology ; Centrosome/physiology ; Dyneins/physiology ; Microtubule-Organizing Center/physiology ; Oocytes/physiology ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {While bacterial symbionts influence a variety of host cellular responses throughout development, there are no documented instances in which symbionts influence early embryogenesis. Here we demonstrate that Wolbachia, an obligate endosymbiont of the parasitic filarial nematodes, is required for proper anterior-posterior polarity establishment in the filarial nematode B. malayi. Characterization of pre- and post-fertilization events in B. malayi reveals that, unlike C. elegans, the centrosomes are maternally derived and produce a cortical-based microtubule organizing center prior to fertilization. We establish that Wolbachia rely on these cortical microtubules and dynein to concentrate at the posterior cortex. Wolbachia also rely on PAR-1 and PAR-3 polarity cues for normal concentration at the posterior cortex. Finally, we demonstrate that Wolbachia depletion results in distinct anterior-posterior polarity defects. These results provide a striking example of endosymbiont-host co-evolution operating on the core initial developmental event of axis determination.}, } @article {pmid25165636, year = {2014}, author = {Metcalf, JA and Jo, M and Bordenstein, SR and Jaenike, J and Bordenstein, SR}, title = {Recent genome reduction of Wolbachia in Drosophila recens targets phage WO and narrows candidates for reproductive parasitism.}, journal = {PeerJ}, volume = {2}, number = {}, pages = {e529}, pmid = {25165636}, issn = {2167-8359}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; T32 GM007347/GM/NIGMS NIH HHS/United States ; }, abstract = {Wolbachia are maternally transmitted endosymbionts that often alter their arthropod hosts' biology to favor the success of infected females, and they may also serve as a speciation microbe driving reproductive isolation. Two of these host manipulations include killing males outright and reducing offspring survival when infected males mate with uninfected females, a phenomenon known as cytoplasmic incompatibility. Little is known about the mechanisms behind these phenotypes, but interestingly either effect can be caused by the same Wolbachia strain when infecting different hosts. For instance, wRec causes cytoplasmic incompatibility in its native host Drosophila recens and male killing in D. subquinaria. The discovery of prophage WO elements in most arthropod Wolbachia has generated the hypothesis that WO may encode genes involved in these reproductive manipulations. However, PCR screens for the WO minor capsid gene indicated that wRec lacks phage WO. Thus, wRec seemed to provide an example where phage WO is not needed for Wolbachia-induced reproductive manipulation. To enable investigation of the mechanism of phenotype switching in different host backgrounds, and to examine the unexpected absence of phage WO, we sequenced the genome of wRec. Analyses reveal that wRec diverged from wMel approximately 350,000 years ago, mainly by genome reduction in the phage regions. While it lost the minor capsid gene used in standard PCR screens for phage WO, it retained two regions encompassing 33 genes, several of which have previously been associated with reproductive parasitism. Thus, WO gene involvement in reproductive manipulation cannot be excluded and reliance on single gene PCR should not be used to rule out the presence of phage WO in Wolbachia. Additionally, the genome sequence for wRec will enable transcriptomic and proteomic studies that may help elucidate the Wolbachia mechanisms of altered reproductive manipulations associated with host switching, perhaps among the 33 remaining phage genes.}, } @article {pmid25158106, year = {2014}, author = {Zhang, G and Hussain, M and Asgari, S}, title = {Regulation of arginine methyltransferase 3 by a Wolbachia-induced microRNA in Aedes aegypti and its effect on Wolbachia and dengue virus replication.}, journal = {Insect biochemistry and molecular biology}, volume = {53}, number = {}, pages = {81-88}, doi = {10.1016/j.ibmb.2014.08.003}, pmid = {25158106}, issn = {1879-0240}, mesh = {Aedes/genetics/*microbiology/*virology ; Animals ; Cell Line ; Dengue/*microbiology/transmission/virology ; Dengue Virus/*physiology ; Gene Expression Regulation/genetics ; Insect Vectors/microbiology/virology ; MicroRNAs/genetics ; Protein-Arginine N-Methyltransferases/*genetics/metabolism ; Symbiosis/physiology ; Virus Replication/physiology ; Wolbachia/growth & development/physiology/*virology ; }, abstract = {The gram-negative endosymbiotic bacteria, Wolbachia, have been found to colonize a wide range of invertebrates, including over 40% of insect species. Best known for host reproductive manipulations, some strains of Wolbachia have been shown to reduce the host life span by about 50% and inhibit replication and transmission of dengue virus (DENV) in the mosquito vector, Aedes aegypti. The molecular mechanisms underlying these effects still are not well understood. Our previous studies showed that Wolbachia uses host microRNAs (miRNAs) to manipulate host gene expression for its efficient maintenance and limiting replication of DENV in Ae. aegypti. Protein arginine methyltransferases are structurally and functionally conserved proteins from yeast to human. In mammals, it has been reported that protein arginine methyltransferases such as PRMT1, 5 and 6 could regulate replication of different viruses. Ae. aegypti contains eight members of protein arginine methyltransferases (AaArgM1-8). Here, we show that the wMelPop strain of Wolbachia introduced into Ae. aegypti significantly induces the expression of AaArgM3. Interestingly, we found that Wolbachia uses aae-miR-2940, which is highly upregulated in Wolbachia-infected mosquitoes, to upregulate the expression of AaArgM3. Silencing of AaArgM3 in a mosquito cell line led to a significant reduction in Wolbachia replication, but had no effect on the replication of DENV. These results provide further evidence that Wolbachia uses the host miRNAs to manipulate host gene expression and facilitate colonization in Ae. aegypti mosquito.}, } @article {pmid25156506, year = {2014}, author = {Hamm, CA and Begun, DJ and Vo, A and Smith, CC and Saelao, P and Shaver, AO and Jaenike, J and Turelli, M}, title = {Wolbachia do not live by reproductive manipulation alone: infection polymorphism in Drosophila suzukii and D. subpulchrella.}, journal = {Molecular ecology}, volume = {23}, number = {19}, pages = {4871-4885}, pmid = {25156506}, issn = {1365-294X}, support = {R01 GM084056/GM/NIGMS NIH HHS/United States ; R01 GM104325/GM/NIGMS NIH HHS/United States ; R01GM084056/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Typing Techniques ; DNA, Mitochondrial/genetics ; Drosophila/classification/*microbiology/physiology ; Female ; Fertility ; Male ; Multilocus Sequence Typing ; Phylogeny ; Reproduction ; Wolbachia/classification/*physiology ; }, abstract = {Drosophila suzukii recently invaded North America and Europe. Populations in Hawaii, California, New York and Nova Scotia are polymorphic for Wolbachia, typically with <20% infection frequency. The Wolbachia in D. suzukii, denoted wSuz, is closely related to wRi, the variant prevalent in continental populations of D. simulans. wSuz is also nearly identical to Wolbachia found in D. subpulchrella, plausibly D. suzukii's sister species. This suggests vertical Wolbachia transmission through cladogenesis ('cladogenic transmission'). The widespread occurrence of 7-20% infection frequencies indicates a stable polymorphism. wSuz is imperfectly maternally transmitted, with wild infected females producing on average 5-10% uninfected progeny. As expected from its low frequency, wSuz produces no cytoplasmic incompatibility (CI), that is, no increased embryo mortality when infected males mate with uninfected females, and no appreciable sex-ratio distortion. The persistence of wSuz despite imperfect maternal transmission suggests positive fitness effects. Assuming a balance between selection and imperfect transmission, we expect a fitness advantage on the order of 20%. Unexpectedly, Wolbachia-infected females produce fewer progeny than do uninfected females. We do not yet understand the maintenance of wSuz in D. suzukii. The absence of detectable CI in D. suzukii and D. subpulchrella makes it unlikely that CI-based mechanisms could be used to control this species without transinfection using novel Wolbachia. Contrary to their reputation as horizontally transmitted reproductive parasites, many Wolbachia infections are acquired through introgression or cladogenesis and many cause no appreciable reproductive manipulation. Such infections, likely to be mutualistic, may be central to understanding the pervasiveness of Wolbachia among arthropods.}, } @article {pmid25155417, year = {2014}, author = {Baldridge, GD and Baldridge, AS and Witthuhn, BA and Higgins, L and Markowski, TW and Fallon, AM}, title = {Proteomic profiling of a robust Wolbachia infection in an Aedes albopictus mosquito cell line.}, journal = {Molecular microbiology}, volume = {94}, number = {3}, pages = {537-556}, pmid = {25155417}, issn = {1365-2958}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; 5 R01 AI 081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes ; Animals ; Bacterial Proteins/*analysis ; Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Proteome/*analysis ; Tandem Mass Spectrometry ; Wolbachia/*growth & development ; }, abstract = {Wolbachia pipientis, a widespread vertically transmitted intracellular bacterium, provides a tool for insect control through manipulation of host-microbe interactions. We report proteomic characterization of wStr, a Wolbachia strain associated with a strong cytoplasmic incompatibility phenotype in its native host, Laodelphax striatellus. In the Aedes albopictus C/wStr1 mosquito cell line, wStr maintains a robust, persistent infection. MS/MS analyses of gel bands revealed a protein 'footprint' dominated by Wolbachia-encoded chaperones, stress response and cell membrane proteins, including the surface antigen WspA, a peptidoglycan-associated lipoprotein and a 73 kDa outer membrane protein. Functional classifications and estimated abundance levels of 790 identified proteins suggested that expression, stabilization and secretion of proteins predominate over bacterial genome replication and cell division. High relative abundances of cysteine desulphurase, serine/glycine hydroxymethyl transferase, and components of the α-ketoglutarate dehydrogenase complex in conjunction with above average abundances of glutamate dehydrogenase and proline utilization protein A support Wolbachia genome-based predictions for amino acid metabolism as a primary energy source. wStr expresses 15 Vir proteins of a Type IV secretion system and its transcriptional regulator. Proteomic characterization of a robust insect-associated Wolbachia strain provides baseline information that will inform further development of in vitro protocols for Wolbachia manipulation.}, } @article {pmid25154109, year = {2014}, author = {Rasić, G and Endersby, NM and Williams, C and Hoffmann, AA}, title = {Using Wolbachia-based release for suppression of Aedes mosquitoes: insights from genetic data and population simulations.}, journal = {Ecological applications : a publication of the Ecological Society of America}, volume = {24}, number = {5}, pages = {1226-1234}, doi = {10.1890/13-1305.1}, pmid = {25154109}, issn = {1051-0761}, mesh = {*Aedes ; Animals ; Australia ; Insect Control/*methods ; Insect Vectors ; Queensland ; *Wolbachia ; }, abstract = {A novel strategy for suppressing disease transmission by Aedes aegypti, the main vector of dengue, uses releases of mosquitoes infected with the bacterium Wolbachia pipientis. Wolbachia are currently released to interfere with viral transmission, but there is also potential to use strains in mosquito suppression and elimination programs via the deleterious effects of the bacterium on the host. Mosquito suppression depends on target areas being relatively isolated to prevent reinvasion and on local climatic conditions. Here we explored the opportunity for suppression of A. aegypti in central Queensland, Australia, by using microsatellite data and simulations based on CIMSiM models of local weather conditions and breeding container data. Our results indicate that Wolbachia-induced extinctions in central Queensland are possible, although they may eventually be compromised by ongoing mosquito migration between towns until these sources are also suppressed. The results highlight a novel use of deleterious Wolbachia infections to achieve ecological as well as disease-related endpoints.}, } @article {pmid25146840, year = {2015}, author = {Ortiz, MF and Wallau, GL and Graichen, DÂ and Loreto, EL}, title = {An evaluation of the ecological relationship between Drosophila species and their parasitoid wasps as an opportunity for horizontal transposon transfer.}, journal = {Molecular genetics and genomics : MGG}, volume = {290}, number = {1}, pages = {67-78}, pmid = {25146840}, issn = {1617-4623}, mesh = {Animals ; Base Sequence ; DNA Transposable Elements/*genetics ; Drosophila/microbiology/*parasitology ; Gene Transfer, Horizontal/*genetics ; Genes, Mitochondrial ; Genome, Insect/genetics ; *Host-Parasite Interactions ; Phylogeny ; Reproducibility of Results ; Species Specificity ; Wasps/*physiology/virology ; Wolbachia/physiology ; }, abstract = {Evidences of horizontal transfer, the exchange of genetic material between reproductively isolated species, have accumulated over the last decades, including for multicellular eukaryotic organisms. However, the mechanisms and ecological relationships that promote such phenomenon is still poorly known. Host-parasite interaction is one type of relationship usually pointed in the literature that could potentially increase the probability of the horizontal transfer between species, because the species involved in such relationships are generally in close contact. Transposable elements, which are well-known genomic parasites, are DNA entities that tend to be involved in horizontal transfer due to their ability to mobilize between different genomic locations. Using Drosophila species and their parasitoid wasps as a host-parasite model, we evaluated the hypothesis that horizontal transposon transfers (HTTs) are more frequent in this set of species than in species that do not exhibit a close ecological and phylogenetic relationship. For this purpose, we sequenced two sets of species using a metagenomic and single-species genomic sampling approach through next-generation DNA sequencing. The first set was composed of five generalist Drosophila (D. maculifrons, D. bandeirantorum, D. polymorpha, D. mercatorum and D. willistoni) species and their associated parasitoid wasps, whereas the second set was composed of D. incompta, which is a flower specialist species, and its parasitoid wasp. We did not find strong evidence of HTT in the two sets of Drosophila and wasp parasites. However, at least five cases of HTT were observed between the generalist and specialist Drosophila species. Moreover, we detected an HT event involving a Wolbachia lineage between generalist and specialist species, indicating that these endosymbiotic bacteria could play a role as HTT vectors. In summary, our results do not support the hypothesis of prevalent HTT between species with a host-parasite relationship, at least for the studied wasp-Drosophila pairs. Moreover, it suggests that other mechanisms or parasites are involved in promoting HTT between Drosophila species as the Wolbachia endosymbiotic bacteria.}, } @article {pmid25132534, year = {2014}, author = {Granquist, EG and Kristiansson, M and Lindgren, PE and Matussek, A and Nødtvedt, A and Okstad, W and Stuen, S}, title = {Evaluation of microbial communities and symbionts in Ixodes ricinus and ungulate hosts (Cervus elaphus and Ovis aries) from shared habitats on the west coast of Norway.}, journal = {Ticks and tick-borne diseases}, volume = {5}, number = {6}, pages = {780-784}, doi = {10.1016/j.ttbdis.2014.05.005}, pmid = {25132534}, issn = {1877-9603}, mesh = {Anaplasma phagocytophilum/physiology ; Animals ; Arachnid Vectors/*microbiology ; Borrelia/genetics/isolation & purification ; Coinfection ; Deer ; Ecosystem ; Female ; Humans ; Ixodes/*microbiology ; Male ; Norway/epidemiology ; Nymph ; Rickettsia/genetics/*isolation & purification ; Sheep ; Sheep Diseases/*epidemiology/microbiology ; Tick-Borne Diseases/*epidemiology/microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Recent reports suggest a potential for transmission of a newly discovered rickettsial endosymbiont, Midichloria mitochondrii, to animals and humans from feeding ticks (Ixodes ricinus). Using molecular methods; I. ricinus, sheep and red deer in Anaplasma phagocytophilum-endemic areas of Norway, were examined to see if they were infected by M. mitochondrii or related organisms like Wolbachia pipientis and Rickettsia spp. A total of 532 ticks collected from pastures, 76 blood samples from grazing lambs and 12 organ samples from hunted deer, were analyzed during the study. All larval pools, 60.4% pooled nymphs and 35.1% of adult ticks were positive for M. mitochondrii. There was a significant difference between geographical areas in the prevalence of M. mitochondrii infection among nymphs. A total of 2.2% pooled nymphs and 5.3% adult ticks were positive for A. phagocytophilum. Eleven percent of pooled nymphs were positive for Borrelia spp, 2.2% of pooled nymphs and 3.5% of adult ticks were positive for Rickettsia spp. and none of the ticks were positive for W. pipientis. The prevalence of A. phagocytophilum infection was 54% and 75% in grazing lambs and deer, respectively. No animals were positive for Borrelia spp., M. mitochondrii, Rickettsia spp. or W. pipientis. The reported findings suggest that M. mitochondrii is widespread in tick populations at different geographical sites, and may appear in co-infection with A. phagocytophilum, Borrelia spp. and Rickettsia spp. in ticks.}, } @article {pmid25121762, year = {2014}, author = {Pessoa, D and Souto-Maior, C and Gjini, E and Lopes, JS and Ceña, B and Codeço, CT and Gomes, MG}, title = {Unveiling time in dose-response models to infer host susceptibility to pathogens.}, journal = {PLoS computational biology}, volume = {10}, number = {8}, pages = {e1003773}, pmid = {25121762}, issn = {1553-7358}, mesh = {Animals ; Dicistroviridae/pathogenicity ; *Disease Susceptibility/microbiology/physiopathology/virology ; *Drosophila melanogaster/microbiology/physiology/virology ; Host-Pathogen Interactions/*physiology ; Male ; *Models, Biological ; Survival Analysis ; Symbiosis/physiology ; Wolbachia/physiology ; }, abstract = {The biological effects of interventions to control infectious diseases typically depend on the intensity of pathogen challenge. As much as the levels of natural pathogen circulation vary over time and geographical location, the development of invariant efficacy measures is of major importance, even if only indirectly inferrable. Here a method is introduced to assess host susceptibility to pathogens, and applied to a detailed dataset generated by challenging groups of insect hosts (Drosophila melanogaster) with a range of pathogen (Drosophila C Virus) doses and recording survival over time. The experiment was replicated for flies carrying the Wolbachia symbiont, which is known to reduce host susceptibility to viral infections. The entire dataset is fitted by a novel quantitative framework that significantly extends classical methods for microbial risk assessment and provides accurate distributions of symbiont-induced protection. More generally, our data-driven modeling procedure provides novel insights for study design and analyses to assess interventions.}, } @article {pmid25114252, year = {2014}, author = {Hughes, GL and Dodson, BL and Johnson, RM and Murdock, CC and Tsujimoto, H and Suzuki, Y and Patt, AA and Cui, L and Nossa, CW and Barry, RM and Sakamoto, JM and Hornett, EA and Rasgon, JL}, title = {Native microbiome impedes vertical transmission of Wolbachia in Anopheles mosquitoes.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {111}, number = {34}, pages = {12498-12503}, pmid = {25114252}, issn = {1091-6490}, support = {R21 AI111175/AI/NIAID NIH HHS/United States ; R21 AI070178/AI/NIAID NIH HHS/United States ; R01 AI067371/AI/NIAID NIH HHS/United States ; R01AI067371/AI/NIAID NIH HHS/United States ; R21AI111175/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Acetobacteraceae/drug effects/growth & development ; Animals ; Anopheles/*microbiology ; Anti-Bacterial Agents/pharmacology ; Biological Evolution ; Disease Transmission, Infectious ; Female ; Infectious Disease Transmission, Vertical ; Microbiota/drug effects ; Ovum/microbiology ; Symbiosis ; Wolbachia/*growth & development ; }, abstract = {Over evolutionary time, Wolbachia has been repeatedly transferred between host species contributing to the widespread distribution of the symbiont in arthropods. For novel infections to be maintained, Wolbachia must infect the female germ line after being acquired by horizontal transfer. Although mechanistic examples of horizontal transfer exist, there is a poor understanding of factors that lead to successful vertical maintenance of the acquired infection. Using Anopheles mosquitoes (which are naturally uninfected by Wolbachia) we demonstrate that the native mosquito microbiota is a major barrier to vertical transmission of a horizontally acquired Wolbachia infection. After injection into adult Anopheles gambiae, some strains of Wolbachia invade the germ line, but are poorly transmitted to the next generation. In Anopheles stephensi, Wolbachia infection elicited massive blood meal-induced mortality, preventing development of progeny. Manipulation of the mosquito microbiota by antibiotic treatment resulted in perfect maternal transmission at significantly elevated titers of the wAlbB Wolbachia strain in A. gambiae, and alleviated blood meal-induced mortality in A. stephensi enabling production of Wolbachia-infected offspring. Microbiome analysis using high-throughput sequencing identified that the bacterium Asaia was significantly reduced by antibiotic treatment in both mosquito species. Supplementation of an antibiotic-resistant mutant of Asaia to antibiotic-treated mosquitoes completely inhibited Wolbachia transmission and partly contributed to blood meal-induced mortality. These data suggest that the components of the native mosquito microbiota can impede Wolbachia transmission in Anopheles. Incompatibility between the microbiota and Wolbachia may in part explain why some hosts are uninfected by this endosymbiont in nature.}, } @article {pmid25108130, year = {2014}, author = {Lefoulon, E and Kuzmin, Y and Plantard, O and Mutafchiev, Y and Otranto, D and Martin, C and Bain, O}, title = {Redescription of Cercopithifilaria rugosicauda (Böhm & Supperer, 1953) (Spirurida: Onchocercidae) of roe deer, with an emended diagnosis of the genus Cercopithifilaria and a genetic characterisation.}, journal = {Parasitology international}, volume = {63}, number = {6}, pages = {808-816}, doi = {10.1016/j.parint.2014.07.011}, pmid = {25108130}, issn = {1873-0329}, mesh = {Animals ; Base Sequence ; DNA, Helminth/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Deer/*parasitology ; Filariasis/*diagnosis/parasitology ; Filarioidea/anatomy & histology/*classification/cytology/genetics ; Male ; Microfilariae ; Molecular Sequence Data ; Sequence Analysis, DNA ; Wolbachia/genetics/isolation & purification ; }, abstract = {Newly collected material of Cercopithifilaria rugosicauda from roe deer Capreolus capreolus was analysed and compared to descriptions of C. rugosicauda from Austria and Hungary. The present specimens were assigned to the genus Cercopithifilaria using both morphological and molecular analyses. Complementary morphological data on the males and microfilariae of C. rugosicauda were described. The main morphological characters of different species of Cercopithifilaria were outlined and an emended generic diagnosis proposed. A genetic characterisation based on the analyses of cox1 and 12S rDNA sequences was reported supporting that C. rugosicauda was included in the clade of the genus Cercopithifilaria distinctly from other congeneric species available. However, these molecular analyses did not solve the relationships between the species of Cercopithifilaria. These could be approached using morphological characters that might be representative of their evolutionary history. In addition, Wolbachia was not seen in C. rugosicauda, either by immunohistological or by molecular approaches.}, } @article {pmid25108053, year = {2014}, author = {Pigeault, R and Braquart-Varnier, C and Marcadé, I and Mappa, G and Mottin, E and Sicard, M}, title = {Modulation of host immunity and reproduction by horizontally acquired Wolbachia.}, journal = {Journal of insect physiology}, volume = {70}, number = {}, pages = {125-133}, doi = {10.1016/j.jinsphys.2014.07.005}, pmid = {25108053}, issn = {1879-1611}, mesh = {Animals ; Female ; Isopoda/immunology/*microbiology/physiology ; Male ; Phagocytosis/physiology ; Reproduction/physiology ; Symbiosis/immunology/physiology ; Wolbachia/*immunology ; }, abstract = {The Wolbachia are symbiotic bacteria vertically transmitted from one host generation to another. However, a growing amount of data shows that horizontal transfers of Wolbachia also frequently occur within and between host species. The consequences of the arrival of new symbionts on host physiology can be studied by their experimental introduction in asymbiotic hosts. After experimental transfers of the eight major isopod Wolbachia strains in the isopod Porcellio dilatatus only two of them (wCon and wDil) were found to (1) have no pathogenic effect on the host and (2) be able to pass vertically to the host offspring. In the present work, we studied the influence of these two strains, able to complete an horizontal transfer, on immunity and reproduction of P. dilatatus at two stages of the transfer: (1) in recipient hosts that encounter the symbionts: to test the influence of symbiont when acquired during host life and (2) in vertically infected offspring: to test the influence of a symbiotic interaction occurring all lifelong. The impact of Wolbachia varied depending on the stage: there were clearer effects in vertically infected individuals than in those that acquired the symbionts during their lives. Moreover, the two Wolbachia strains showed contrasted effects: the strain wCon tended to reduce the reproductive investment but to maintain or increase immune parameters whilst wDil had positive effects on reproductive investment but decreased the investment in some immune parameters. These results suggest that horizontally acquisition of Wolbachia can influence the balance between host immune and reproductive traits.}, } @article {pmid25096321, year = {2014}, author = {Leichty, AR and Brisson, D}, title = {Selective whole genome amplification for resequencing target microbial species from complex natural samples.}, journal = {Genetics}, volume = {198}, number = {2}, pages = {473-481}, pmid = {25096321}, issn = {1943-2631}, support = {AI076342/AI/NIAID NIH HHS/United States ; T32 GM008216/GM/NIGMS NIH HHS/United States ; R01 AI091595/AI/NIAID NIH HHS/United States ; R01 AI076342/AI/NIAID NIH HHS/United States ; R01 AI097137/AI/NIAID NIH HHS/United States ; T32GM008216,/GM/NIGMS NIH HHS/United States ; AI097137/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Borrelia burgdorferi/genetics ; DNA, Bacterial/genetics ; Drosophila melanogaster/genetics ; *Environmental Microbiology ; Escherichia coli/genetics ; Female ; Gene Library ; Genome, Bacterial ; Nucleic Acid Amplification Techniques ; *Sequence Analysis, DNA ; }, abstract = {Population genomic analyses have demonstrated power to address major questions in evolutionary and molecular microbiology. Collecting populations of genomes is hindered in many microbial species by the absence of a cost effective and practical method to collect ample quantities of sufficiently pure genomic DNA for next-generation sequencing. Here we present a simple method to amplify genomes of a target microbial species present in a complex, natural sample. The selective whole genome amplification (SWGA) technique amplifies target genomes using nucleotide sequence motifs that are common in the target microbe genome, but rare in the background genomes, to prime the highly processive phi29 polymerase. SWGA thus selectively amplifies the target genome from samples in which it originally represented a minor fraction of the total DNA. The post-SWGA samples are enriched in target genomic DNA, which are ideal for population resequencing. We demonstrate the efficacy of SWGA using both laboratory-prepared mixtures of cultured microbes as well as a natural host-microbe association. Targeted amplification of Borrelia burgdorferi mixed with Escherichia coli at genome ratios of 1:2000 resulted in >10(5)-fold amplification of the target genomes with <6.7-fold amplification of the background. SWGA-treated genomic extracts from Wolbachia pipientis-infected Drosophila melanogaster resulted in up to 70% of high-throughput resequencing reads mapping to the W. pipientis genome. By contrast, 2-9% of sequencing reads were derived from W. pipientis without prior amplification. The SWGA technique results in high sequencing coverage at a fraction of the sequencing effort, thus allowing population genomic studies at affordable costs.}, } @article {pmid25091123, year = {2015}, author = {Suh, E and Sim, C and Park, JJ and Cho, K}, title = {Inter-population variation for Wolbachia induced reproductive incompatibility in the haplodiploid mite Tetranychus urticae.}, journal = {Experimental & applied acarology}, volume = {65}, number = {1}, pages = {55-71}, pmid = {25091123}, issn = {1572-9702}, mesh = {Animal Distribution ; Animals ; Cloning, Molecular ; DNA, Bacterial/genetics ; Female ; Male ; Phylogeny ; Ploidies ; Polymerase Chain Reaction ; Reproduction ; Republic of Korea ; Tetranychidae/genetics/*microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Recent studies have revealed diverse patterns of cytoplasmic incompatibility (CI) induced by Wolbachia in the two spotted spider mite (Tetranychus urticae Koch). The mechanism of CI consists of two steps: modification (mod) of sperm of infected males and the rescue (resc) of these chromosomes by Wolbachia in the egg, which results in female embryonic mortality (FM), male development (MD) or no CI. Our study reports that Wolbachia infections were highly prevalent infecting all T. urticae populations from various crops in 14 commercial greenhouses in Korea, with two Wolbachia strains expressing distinctive phenotypic effects on hosts. Analyses for wsp gene sequences obtained from collected mite populations revealed all sequences were categorized into two groups (group W1 and W2) discriminated by three diagnostic nucleotides while all Wolbachia strains belonged to the subgroup Ori in Wolbachia supergroup B. Host plants of each mite population were also generally correlated this grouping. Various mating experiments with two mite populations from each group showed that CI patterns and host plants of the mite populations were completely matched with the grouping; no CI (mod(-)resc(+)) for group W1 and mixed pattern of FM and MD (mod(+)resc(+)) for group W2. No distinct changes in fecundity or sex ratio due to Wolbachia infections were observed in four mite populations regardless of Wolbachia grouping. Our study suggests a potential correlation between phenotypic effect of Wolbachia infection and its genetic diversity associated with host plants in Korean mite populations.}, } @article {pmid25077993, year = {2014}, author = {Jiang, W and Zhu, J and Chen, M and Yang, Q and Du, X and Chen, S and Zhang, L and Yu, Y and Yu, W}, title = {Wolbachia infection status and genetic structure in natural populations of Polytremis nascens (Lepidoptera: Hesperiidae).}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {27}, number = {}, pages = {202-211}, doi = {10.1016/j.meegid.2014.07.026}, pmid = {25077993}, issn = {1567-7257}, mesh = {Animals ; China ; DNA, Mitochondrial ; Evolution, Molecular ; Genetic Structures ; Genetic Variation ; Genetics, Population ; Geography ; Host-Pathogen Interactions/genetics ; Lepidoptera/classification/*genetics/*microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S ; *Wolbachia ; }, abstract = {The maternally inherited obligate bacteria Wolbachia is known for infecting the reproductive tissues of a wide range of arthropods. In this study, we surveyed Wolbachia infections in Polytremis nascens (Lepidoptera: Hesperiidae) from 14 locations in China by amplifying the 16S rRNA gene with a nested PCR method and revealed the effect of Wolbachia on host mitochondrial DNA. The results show that 31% (21/67) are Wolbachia positive among all specimens and mainly prevails in southern populations in China. No significant difference in the prevalence is found between the sexes. Notably, the nucleotide diversity of Wolbachia infected butterflies is smaller compared to that of uninfected butterflies. The mitochondrial DNA of infected group appear to be not evolving neutrally (Tajima's D value=-2.3303 and Fu's F values=-3.7068). The analysis of molecular variance shows significant differentiation of mitochondrial haplotypes between infected and uninfected specimens (FST=0.6064). The mismatch analysis speculated the different expansion pattern in Wolbachia infected specimens and all P. nascens specimens. These results suggest that the populations of P. nascens may have recently been subjected to a Wolbachia-induced sweep. Additionally, phylogenetic analysis differentiated the mitochondrial haplotypes of P. nascens into three major clades. The clades are in perfect agreement with the pattern of Wolbachia infection. One of the clades grouped with the butterflies infected with Wolbachia. The remaining two clades grouped with uninfected butterflies from the central-west of China populations and Eastern and Southern China populations respectively, which are isolated mainly by the Yangtze River. The analysis of haplotype networks, geographic distribution and population size change shows that Haplotype 1 in central-west of China is the ancestral haplotype and the populations of P. nascens are expanded.}, } @article {pmid25077022, year = {2014}, author = {Bing, XL and Xia, WQ and Gui, JD and Yan, GH and Wang, XW and Liu, SS}, title = {Diversity and evolution of the Wolbachia endosymbionts of Bemisia (Hemiptera: Aleyrodidae) whiteflies.}, journal = {Ecology and evolution}, volume = {4}, number = {13}, pages = {2714-2737}, pmid = {25077022}, issn = {2045-7758}, abstract = {Wolbachia is the most prevalent symbiont described in arthropods to date. Wolbachia can manipulate host reproduction, provide nutrition to insect hosts and protect insect hosts from pathogenic viruses. So far, 13 supergroups of Wolbachia have been identified. The whitefly Bemisia tabaci is a complex containing more than 28 morphologically indistinguishable cryptic species. Some cryptic species of this complex are invasive. In this study, we report a comprehensive survey of Wolbachia in B. tabaci and its relative B. afer from 1658 insects representing 54 populations across 13 provinces of China and one state of Australia. Based on the results of PCR or sequencing of the 16S rRNA gene, the overall rates of Wolbachia infection were 79.6% and 0.96% in the indigenous and invasive Bemisia whiteflies, respectively. We detected a new Wolbachia supergroup by sequencing five molecular marker genes including 16S rRNA, groEL, gltA, hcpA, and fbpA genes. Data showed that many protein-coding genes have limitations in detecting and classifying newly identified Wolbachia supergroups and thus raise a challenge to the known Wolbachia MLST standard analysis system. Besides, the other Wolbachia strains detected from whiteflies were clustered into supergroup B. Phylogenetic trees of whitefly mitochondrial cytochrome oxidase subunit I and Wolbachia multiple sequencing typing genes were not congruent. In addition, Wolbachia was also detected outside the special bacteriocytes in two cryptic species by fluorescence in situ hybridization, indicating the horizontal transmission of Wolbachia. Our results indicate that members of Wolbachia are far from well explored.}, } @article {pmid25072029, year = {2014}, author = {Pruneau, L and Moumène, A and Meyer, DF and Marcelino, I and Lefrançois, T and Vachiéry, N}, title = {Understanding Anaplasmataceae pathogenesis using "Omics" approaches.}, journal = {Frontiers in cellular and infection microbiology}, volume = {4}, number = {}, pages = {86}, pmid = {25072029}, issn = {2235-2988}, mesh = {Anaplasmataceae/*physiology ; Anaplasmataceae Infections/*etiology ; Animals ; Gene Expression Profiling ; *Genomics/methods ; Host-Pathogen Interactions ; Humans ; *Proteomics/methods ; Ticks/microbiology ; Transcriptome ; }, abstract = {This paper examines how "Omics" approaches improve our understanding of Anaplasmataceae pathogenesis, through a global and integrative strategy to identify genes and proteins involved in biochemical pathways key for pathogen-host-vector interactions. The Anaplasmataceae family comprises obligate intracellular bacteria mainly transmitted by arthropods. These bacteria are responsible for major human and animal endemic and emerging infectious diseases with important economic and public health impacts. In order to improve disease control strategies, it is essential to better understand their pathogenesis. Our work focused on four Anaplasmataceae, which cause important animal, human and zoonotic diseases: Anaplasma marginale, A. phagocytophilum, Ehrlichia chaffeensis, and E. ruminantium. Wolbachia spp. an endosymbiont of arthropods was also included in this review as a model of a non-pathogenic Anaplasmataceae. A gap analysis on "Omics" approaches on Anaplasmataceae was performed, which highlighted a lack of studies on the genes and proteins involved in the infection of hosts and vectors. Furthermore, most of the studies have been done on the pathogen itself, mainly on infectious free-living forms and rarely on intracellular forms. In order to perform a transcriptomic analysis of the intracellular stage of development, researchers developed methods to enrich bacterial transcripts from infected cells. These methods are described in this paper. Bacterial genes encoding outer membrane proteins, post-translational modifications, eukaryotic repeated motif proteins, proteins involved in osmotic and oxidative stress and hypothetical proteins have been identified to play a key role in Anaplasmataceae pathogenesis. Further investigations on the function of these outer membrane proteins and hypothetical proteins will be essential to confirm their role in the pathogenesis. Our work underlines the need for further studies in this domain and on host and vector responses to infection.}, } @article {pmid25062354, year = {2014}, author = {Caspi-Fluger, A and Inbar, M and Steinberg, S and Friedmann, Y and Freund, M and Mozes-Daube, N and Zchori-Fein, E}, title = {Characterization of the symbiont Rickettsia in the mirid bug Nesidiocoris tenuis (Reuter) (Heteroptera: Miridae).}, journal = {Bulletin of entomological research}, volume = {104}, number = {6}, pages = {681-688}, doi = {10.1017/S0007485314000492}, pmid = {25062354}, issn = {1475-2670}, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Female ; Heteroptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Molecular Sequence Data ; RNA, Ribosomal, 16S ; Real-Time Polymerase Chain Reaction ; Rickettsia/genetics/metabolism/*physiology ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Nesidiocoris tenuis (Reuter) (Heteroptera: Miridae) is an omnivorous insect used for biological control. Augmentative release and conservation of N. tenuis have been used for pest control in tomato crops. Intracellular bacterial symbionts of arthropods are common in nature and have diverse effects on their hosts; in some cases they can dramatically affect biological control. Fingerprinting methods showed that the symbiotic complex associated with N. tenuis includes Wolbachia and Rickettsia. Rickettsia of N. tenuis was further characterized by sequencing the 16S rRNA and gltA bacterial genes, measuring its amount in different developmental stages of the insect by real-time polymerase chain reaction, and localizing the bacteria in the insect's body by fluorescence in situ hybridization. The Rickettsia in N. tenuis exhibited 99 and 96% similarity of both sequenced genes to Rickettsia bellii and Rickettsia reported from Bemisia tabaci, respectively. The highest amount of Rickettsia was measured in the 5th instar and adult, and the symbionts could be detected in the host gut and ovaries. Although the role played by Rickettsia in the biology of N. tenuis is currently unknown, their high amount in the adults and localization in the gut suggest that they may have a nutritional role in this insect.}, } @article {pmid25059495, year = {2015}, author = {Pathak, M and Verma, M and Srivastava, M and Misra-Bhattacharya, S}, title = {Wolbachia endosymbiont of Brugia malayi elicits a T helper type 17-mediated pro-inflammatory immune response through Wolbachia surface protein.}, journal = {Immunology}, volume = {144}, number = {2}, pages = {231-244}, pmid = {25059495}, issn = {1365-2567}, mesh = {Animals ; Bacterial Outer Membrane Proteins/biosynthesis/genetics/*pharmacology ; Brugia malayi/*immunology/*microbiology ; Cloning, Molecular ; Filariasis/immunology/*microbiology ; Inflammation/immunology ; Interferon-gamma/biosynthesis ; Interleukin-10/biosynthesis ; Interleukin-2/biosynthesis ; Interleukin-4/biosynthesis ; Larva ; Lymph Nodes/immunology/microbiology/parasitology ; Male ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins/biosynthesis/genetics/pharmacology ; Spleen/immunology/microbiology/parasitology ; T-Lymphocytes, Regulatory/*immunology ; Th17 Cells/*immunology ; Th2 Cells/immunology ; Transforming Growth Factor beta/biosynthesis ; Wolbachia/*immunology ; }, abstract = {Wolbachia is an endosymbiotic bacterium of the filarial nematode Brugia malayi. The symbiotic relationship between Wolbachia and its filarial host is dependent on interactions between the proteins of both organisms. However, little is known about Wolbachia proteins that are involved in the inflammatory pathology of the host during lymphatic filariasis. In the present study, we cloned, expressed and purified Wolbachia surface protein (r-wsp) from Wolbachia and administered it to mice, either alone or in combination with infective larvae of B. malayi (Bm-L3) and monitored the developing immune response in infected animals. Our results show that spleens and mesenteric lymph nodes of mice immunized with either r-wsp or infected with Bm-L3 show increased percentages of CD4(+) T helper type 17 (Th17) cells and Th1 cytokines like interferon-γ and interleukin-2 (IL-2) along with decreased percentages of regulatory T cells, Th2 cytokines like IL-4 and IL-10 and transforming growth factor β (TGF-β) levels in culture supernatants of splenocytes. These observations were stronger in mice immunized with r-wsp alone. Interestingly, when mice were first immunized with r-wsp and subsequently infected with Bm-L3, percentages of CD4(+) Th17 cells and Th1 cytokines increased even further while that of regulatory T cells, Th2 cytokines and TGF-β levels decreased. These results for the first time show that r-wsp acts synergistically with Bm-L3 in promoting a pro-inflammatory response by increasing Th17 cells and at the same time diminishes host immunological tolerance by decreasing regulatory T cells and TGF-β secretion.}, } @article {pmid25054227, year = {2014}, author = {Williams-Newkirk, AJ and Rowe, LA and Mixson-Hayden, TR and Dasch, GA}, title = {Characterization of the bacterial communities of life stages of free living lone star ticks (Amblyomma americanum).}, journal = {PloS one}, volume = {9}, number = {7}, pages = {e102130}, pmid = {25054227}, issn = {1932-6203}, mesh = {Animals ; Arthropod Vectors/genetics/*growth & development/microbiology ; Bacteria/classification/genetics/*growth & development ; DNA Barcoding, Taxonomic/methods ; Ecosystem ; Female ; Humans ; Ixodidae/genetics/*growth & development/microbiology ; *Life Cycle Stages ; Male ; Metagenomics/methods ; Nymph/genetics/growth & development/microbiology ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA/methods ; }, abstract = {The lone star tick (Amblyomma americanum) is an abundant and aggressive biter of humans, domestic animals, and wildlife in the southeastern-central USA and an important vector of several known and suspected zoonotic bacterial pathogens. However, the biological drivers of bacterial community variation in this tick are still poorly defined. Knowing the community context in which tick-borne bacterial pathogens exist and evolve is required to fully understand the ecology and immunobiology of the ticks and to design effective public health and veterinary interventions. We performed a metagenomic survey of the bacterial communities of questing A. americanum and tested 131 individuals (66 nymphs, 24 males, and 41 females) from five sites in three states. Pyrosequencing was performed with barcoded eubacterial primers targeting variable 16S rRNA gene regions 5-3. The bacterial communities were dominated by Rickettsia (likely R. amblyommii) and an obligate Coxiella symbiont, together accounting for 6.7-100% of sequences per tick. DNAs from Midichloria, Borrelia, Wolbachia, Ehrlichia, Pseudomonas, or unidentified Bacillales, Enterobacteriaceae, or Rhizobiales groups were also detected frequently. Wolbachia and Midichloria significantly co-occurred in Georgia (p<0.00001), but not in other states. The significance of the Midichloria-Wolbachia co-occurrence is unknown. Among ticks collected in Georgia, nymphs differed from adults in both the composition (p = 0.002) and structure (p = 0.002) of their bacterial communities. Adults differed only in their community structure (p = 0.002) with males containing more Rickettsia and females containing more Coxiella. Comparisons among adult ticks collected in New York and North Carolina supported the findings from the Georgia collection despite differences in geography, collection date, and sample handling, implying that the differences detected are consistent attributes. The data also suggest that some members of the bacterial community change during the tick life cycle and that some sex-specific attributes may be detectable in nymphs.}, } @article {pmid25052143, year = {2014}, author = {Sicard, M and Dittmer, J and Grève, P and Bouchon, D and Braquart-Varnier, C}, title = {A host as an ecosystem: Wolbachia coping with environmental constraints.}, journal = {Environmental microbiology}, volume = {16}, number = {12}, pages = {3583-3607}, doi = {10.1111/1462-2920.12573}, pmid = {25052143}, issn = {1462-2920}, mesh = {Animals ; Arthropods/genetics/immunology/*microbiology ; Ecosystem ; Environment ; Female ; Gonads/microbiology ; Microbial Interactions ; Nematoda/genetics/immunology/*microbiology ; Phenotype ; *Symbiosis ; Wolbachia/growth & development/*physiology ; }, abstract = {The Wolbachia are intracellular endosymbionts widely distributed among invertebrates. These primarily vertically transmitted α-proteobacteria have been intensively studied during the last decades because of their intriguing interactions with hosts, ranging from reproductive manipulations to mutualism. To optimize their vertical transmission from mother to offspring, the Wolbachia have developed fine-tuned strategies. However, the Wolbachia are not restricted to the female gonads and frequently exhibit wide intra-host distributions. This extensive colonization of somatic organs might be necessary for Wolbachia to develop their diverse extended phenotypes. From an endosymbiont's perspective, the within-host environment potentially presents different environmental constraints. Hence, the Wolbachia have to face different intracellular habitats, their host's immune system as well as other microorganisms co-occurring in the same host individual and sometimes even in the same cell. A means for the Wolbachia to protect themselves from these environmental constraints may be to live 'hidden' in vacuoles within host cells. In this review, we summarize the current knowledge regarding the extent of the Wolbachia pandemic and discuss the various environmental constraints these bacteria may have to face within their 'host ecosystem'. Finally, we identify new avenues for future research to better understand the complexity of Wolbachia's interactions with their intracellular environment.}, } @article {pmid25048456, year = {2014}, author = {Werblow, A and Klimpel, S and Bolius, S and Dorresteijn, AW and Sauer, J and Melaun, C}, title = {Population structure and distribution patterns of the sibling mosquito species Culex pipiens and Culex torrentium (Diptera: Culicidae) reveal different evolutionary paths.}, journal = {PloS one}, volume = {9}, number = {7}, pages = {e102158}, pmid = {25048456}, issn = {1932-6203}, mesh = {Animals ; Biological Evolution ; Culex/*genetics/physiology ; Genetic Variation ; Germany ; Haplotypes ; Phylogeny ; Phylogeography ; Sequence Analysis, DNA ; }, abstract = {Nowadays a number of endemic mosquito species are known to possess vector abilities for various diseases, as e.g. the sibling species Culex pipiens and Culex torrentium. Due to their morphological similarity, ecology, distribution and vector abilities, knowledge about these species' population structure is essential. Culicidae from 25 different sampling sites were collected from March till October 2012. All analyses were performed with aligned cox1 sequences with a total length of 658 bp. Population structure as well as distribution patterns of both species were analysed using molecular methods and different statistical tests like distance based redundancy analysis (dbDRA), analysis of molecular variances (AMOVA) or McDonald & Kreitman test and Tajima's D. Within both species, we could show a genetic variability among the cox1 fragment. The construction of haplotype networks revealed one dominating haplotype for Cx. pipiens, widely distributed within Germany and a more homogeneous pattern for Cx. torrentium. The low genetic differences within Cx. pipiens could be a result of an infection with Wolbachia which can induce a sweep through populations by passively taking the also maternally inherited mtDNA through the population, thereby reducing the mitochondrial diversity as an outcome of reproductive incompatibility. Pairwise population genetic differentiation (FST) ranged significantly from moderate to very great between populations of Cx. pipiens and Cx. torrentium. Analyses of molecular variances revealed for both species that the main genetic variability exists within the populations (Cx. pipiens [88.38%]; Cx. torrentium [66.54%]). Based on a distance based redundancy analysis geographical origin explained a small but significant part of the species' genetic variation. Overall, the results confirm that Cx. pipiens and Cx. torrentium underlie different factors regarding their mitochondrial differentiation, which could be a result of endosymbiosis, dispersal between nearly located populations or human introduction.}, } @article {pmid25046729, year = {2014}, author = {Slatko, BE and Luck, AN and Dobson, SL and Foster, JM}, title = {Wolbachia endosymbionts and human disease control.}, journal = {Molecular and biochemical parasitology}, volume = {195}, number = {2}, pages = {88-95}, doi = {10.1016/j.molbiopara.2014.07.004}, pmid = {25046729}, issn = {1872-9428}, mesh = {Animals ; Filariasis/drug therapy/*parasitology ; Filarioidea/drug effects/*microbiology/physiology ; Humans ; Nematoda/drug effects/*microbiology/physiology ; Nematode Infections/drug therapy/*parasitology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Most human filarial nematode parasites and arthropods are hosts for a bacterial endosymbiont, Wolbachia. In filaria, Wolbachia are required for normal development, fertility and survival, whereas in arthropods, they are largely parasitic and can influence development and reproduction, but are generally not required for host survival. Due to their obligate nature in filarial parasites, Wolbachia have been a target for drug discovery initiatives using several approaches including diversity and focused library screening and genomic sequence analysis. In vitro and in vivo anti-Wolbachia antibiotic treatments have been shown to have adulticidal activity, a long sought goal of filarial parasite drug discovery. In mosquitoes, it has been shown that the presence of Wolbachia can inhibit the transmission of certain viruses, such as Dengue, Chikungunya, Yellow Fever, West Nile, as well as the infectivity of the malaria-causing protozoan, Plasmodium and filarial nematodes. Furthermore, Wolbachia can cause a form of conditional sterility that can be used to suppress populations of mosquitoes and additional medically important insects. Thus Wolbachia, a pandemic endosymbiont offers great potential for elimination of a wide-variety of devastating human diseases.}, } @article {pmid25041943, year = {2014}, author = {Joshi, D and McFadden, MJ and Bevins, D and Zhang, F and Xi, Z}, title = {Wolbachia strain wAlbB confers both fitness costs and benefit on Anopheles stephensi.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {336}, pmid = {25041943}, issn = {1756-3305}, support = {R01AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/genetics/*microbiology/*physiology ; Female ; Fertility/physiology ; Genetic Fitness ; Host-Pathogen Interactions ; Male ; Sex Ratio ; Sexual Behavior, Animal ; Wolbachia/classification/*physiology ; }, abstract = {BACKGROUND: Wolbachia is a maternally transmitted intracellular bacterium that is estimated to infect up to 65% of insect species, but it is not naturally present in Anopheles malaria vectors. Wolbachia-based strategies for malaria vector control can be developed either through population replacement to reduce vectorial capacity or through population suppression to reduce the mosquito population. We have previously generated An. stephensi mosquitoes carrying a stable wAlbB Wolbachia infection and have demonstrated their ability to invade wild-type laboratory populations and confer resistance to Plasmodium on these populations.

METHODS: We assessed wAlbB-associated fitness by comparing the female fecundity, immature development and survivorship, body size, male mating competiveness, and adult longevity of the infected An. stephensi to that of wild-type mosquitoes.

RESULTS: We found that wAlbB reduced female fecundity and caused a minor decrease in male mating competiveness. We also observed that wAlbB increased the life span of both male and female mosquitoes when they were maintained solely on sugar meals; however, there was no impact on the life span of blood-fed females. In addition, wAlbB did not influence either immature development and survivorship or adult body sizes.

CONCLUSIONS: These results provide significant support for developing Wolbachia-based strategies for malaria vector control.}, } @article {pmid25037159, year = {2014}, author = {Dossi, FC and da Silva, EP and Cônsoli, FL}, title = {Population dynamics and growth rates of endosymbionts during Diaphorina citri (Hemiptera, Liviidae) ontogeny.}, journal = {Microbial ecology}, volume = {68}, number = {4}, pages = {881-889}, pmid = {25037159}, issn = {1432-184X}, mesh = {Animals ; Betaproteobacteria/genetics/growth & development/*physiology ; DNA, Bacterial/genetics/metabolism ; Female ; Halomonadaceae/genetics/growth & development/*physiology ; Hemiptera/growth & development/*microbiology ; Male ; Nymph/growth & development/microbiology ; Ovum/growth & development/microbiology ; Population Dynamics ; RNA, Ribosomal, 16S/genetics/metabolism ; Real-Time Polymerase Chain Reaction ; *Symbiosis ; Wolbachia/genetics/growth & development/*physiology ; }, abstract = {The infection density of symbionts is among the major parameters to understand their biological effects in host-endosymbionts interactions. Diaphorina citri harbors two bacteriome-associated bacterial endosymbionts (Candidatus Carsonella ruddii and Candidatus Profftella armatura), besides the intracellular reproductive parasite Wolbachia. In this study, the density dynamics of the three endosymbionts associated with the psyllid D. citri was investigated by real-time quantitative PCR (qPCR) at different developmental stages. Bacterial density was estimated by assessing the copy number of the 16S rRNA gene for Carsonella and Profftella, and of the ftsZ gene for Wolbachia. Analysis revealed a continuous growth of the symbionts during host development. Symbiont growth and rate curves were estimated by the Gompertz equation, which indicated a negative correlation between the degree of symbiont-host specialization and the time to achieve the maximum growth rate (t*). Carsonella densities were significantly lower than those of Profftella at all host developmental stages analyzed, even though they both displayed a similar trend. The growth rates of Wolbachia were similar to those of Carsonella, but Wolbachia was not as abundant. Adult males displayed higher symbiont densities than females. However, females showed a much more pronounced increase in symbiont density as they aged if compared to males, regardless of the incorporation of symbionts into female oocytes and egg laying. The increased density of endosymbionts in aged adults differs from the usual decrease observed during host aging in other insect-symbiont systems.}, } @article {pmid25010200, year = {2014}, author = {Dodson, BL and Hughes, GL and Paul, O and Matacchiero, AC and Kramer, LD and Rasgon, JL}, title = {Wolbachia enhances West Nile virus (WNV) infection in the mosquito Culex tarsalis.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {7}, pages = {e2965}, pmid = {25010200}, issn = {1935-2735}, support = {R21 AI111175/AI/NIAID NIH HHS/United States ; R21 AI070178/AI/NIAID NIH HHS/United States ; R01 AI067371/AI/NIAID NIH HHS/United States ; R01AI067371/AI/NIAID NIH HHS/United States ; R21AI111175/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Culex/genetics/microbiology/physiology/virology ; *Insect Vectors/genetics/microbiology/physiology/virology ; *Mosquito Control/methods/standards ; Symbiosis ; Transcription Factors/genetics ; West Nile Fever/microbiology/*transmission/virology ; *West Nile virus ; *Wolbachia ; }, abstract = {Novel strategies are required to control mosquitoes and the pathogens they transmit. One attractive approach involves maternally inherited endosymbiotic Wolbachia bacteria. After artificial infection with Wolbachia, many mosquitoes become refractory to infection and transmission of diverse pathogens. We evaluated the effects of Wolbachia (wAlbB strain) on infection, dissemination and transmission of West Nile virus (WNV) in the naturally uninfected mosquito Culex tarsalis, which is an important WNV vector in North America. After inoculation into adult female mosquitoes, Wolbachia reached high titers and disseminated widely to numerous tissues including the head, thoracic flight muscles, fat body and ovarian follicles. Contrary to other systems, Wolbachia did not inhibit WNV in this mosquito. Rather, WNV infection rate was significantly higher in Wolbachia-infected mosquitoes compared to controls. Quantitative PCR of selected innate immune genes indicated that REL1 (the activator of the antiviral Toll immune pathway) was down regulated in Wolbachia-infected relative to control mosquitoes. This is the first observation of Wolbachia-induced enhancement of a human pathogen in mosquitoes, suggesting that caution should be applied before releasing Wolbachia-infected insects as part of a vector-borne disease control program.}, } @article {pmid25005355, year = {2014}, author = {Bertrand, C and Janzen, DH and Hallwachs, W and Burns, JM and Gibson, JF and Shokralla, S and Hajibabaei, M}, title = {Mitochondrial and nuclear phylogenetic analysis with Sanger and next-generation sequencing shows that, in Área de Conservación Guanacaste, northwestern Costa Rica, the skipper butterfly named Urbanus belli (family Hesperiidae) comprises three morphologically cryptic species.}, journal = {BMC evolutionary biology}, volume = {14}, number = {}, pages = {153}, pmid = {25005355}, issn = {1471-2148}, mesh = {Animals ; Butterflies/*classification/*genetics/microbiology ; Cell Nucleus/genetics ; Costa Rica ; DNA, Ribosomal Spacer/genetics ; High-Throughput Nucleotide Sequencing ; Mitochondria/genetics ; Phylogeny ; Wolbachia/genetics ; }, abstract = {BACKGROUND: Skipper butterflies (Hesperiidae) are a relatively well-studied family of Lepidoptera. However, a combination of DNA barcodes, morphology, and natural history data has revealed several cryptic species complexes within them. Here, we investigate three DNA barcode lineages of what has been identified as Urbanus belli (Hesperiidae, Eudaminae) in Área de Conservación Guanacaste (ACG), northwestern Costa Rica.

RESULTS: Although no morphological traits appear to distinguish among the three, congruent nuclear and mitochondrial lineage patterns show that "Urbanus belli" in ACG is a complex of three sympatric species. A single strain of Wolbachia present in two of the three cryptic species indicates that Urbanus segnestami Burns (formerly Urbanus belliDHJ01), Urbanus bernikerni Burns (formerly Urbanus belliDHJ02), and Urbanus ehakernae Burns (formerly Urbanus belliDHJ03) may be biologically separated by Wolbachia, as well as by their genetics. Use of parallel sequencing through 454-pyrosequencing improved the utility of ITS2 as a phylogenetic marker and permitted examination of the intra- and interlineage relationships of ITS2 variants within the species complex. Interlineage, intralineage and intragenomic compensatory base pair changes were discovered in the secondary structure of ITS2.

CONCLUSION: These findings corroborate the existence of three cryptic species. Our confirmation of a novel cryptic species complex, initially suggested by DNA barcode lineages, argues for using a multi-marker approach coupled with next-generation sequencing for exploration of other suspected species complexes.}, } @article {pmid24992829, year = {2014}, author = {Kelly-Hope, LA and Cano, J and Stanton, MC and Bockarie, MJ and Molyneux, DH}, title = {Innovative tools for assessing risks for severe adverse events in areas of overlapping Loa loa and other filarial distributions: the application of micro-stratification mapping.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {307}, pmid = {24992829}, issn = {1756-3305}, support = {G1001337/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Democratic Republic of the Congo/epidemiology ; Humans ; *Loa ; Loiasis/*epidemiology/*pathology ; Prevalence ; Risk Factors ; }, abstract = {BACKGROUND: The wide distribution of Loa loa infection (loiasis) throughout the Democratic Republic of Congo (DRC) is a major obstacle to the plans to eliminate onchocerciasis and lymphatic filariasis (LF) because the standard drug regime is dependent on ivermectin, which cannot be used in co-endemic areas due to the risk of severe adverse events (SAEs). A better understanding of the micro-epidemiology, overlapping low and high risk areas, and how they relate to SAEs is critical to ensure safe and effective treatment.

FINDINGS: Based on published data from the Bas Congo Province in DRC, this study used geographical information systems (GIS) to re-map and analyse onchocerciasis and loiasis prevalence (<20%, 20 to 40%, >40%) at 144 sites in relation to health district areas reporting SAEs. The new maps highlighted the contrasting patterns of the high prevalence sites, and significant geographical overlap between low onchocerciasis and high loiasis sites. Statistical analyses found that sites with medium to high loiasis prevalence were 10 to 16 times more likely to be in a SAE area than those with low prevalence of loiasis. Sites where both onchocerciasis and loiasis prevalence was >20% were also associated with SAE areas.

CONCLUSIONS: Collaborative efforts between the national onchocerciasis and LF programmes are critical as plans to scale interventions are moving forward and thus, alternative strategies needed in loiasis co-endemic areas which may include the new L. loa test and treat strategy using the Cellscope, or interventions such as integrated vector management, or anti Wolbachia therapy using doxycycline.}, } @article {pmid24990702, year = {2014}, author = {Panchin, AY and Tuzhikov, AI and Panchin, YV}, title = {Midichlorians--the biomeme hypothesis: is there a microbial component to religious rituals?.}, journal = {Biology direct}, volume = {9}, number = {}, pages = {14}, pmid = {24990702}, issn = {1745-6150}, mesh = {Animals ; Bacteria/*metabolism ; *Ceremonial Behavior ; Humans ; *Models, Biological ; *Religion ; }, abstract = {BACKGROUND: Cutting edge research of human microbiome diversity has led to the development of the microbiome-gut-brain axis concept, based on the idea that gut microbes may have an impact on the behavior of their human hosts. Many examples of behavior-altering parasites are known to affect members of the animal kingdom. Some prominent examples include Ophiocordyceps unilateralis (fungi), Toxoplasma gondii (protista), Wolbachia (bacteria), Glyptapanteles sp. (arthropoda), Spinochordodes tellinii (nematomorpha) and Dicrocoelium dendriticum (flat worm). These organisms belong to a very diverse set of taxonomic groups suggesting that the phenomena of parasitic host control might be more common in nature than currently established and possibly overlooked in humans.

Some microorganisms would gain an evolutionary advantage by encouraging human hosts to perform certain rituals that favor microbial transmission. We hypothesize that certain aspects of religious behavior observed in the human society could be influenced by microbial host control and that the transmission of some religious rituals could be regarded as the simultaneous transmission of both ideas (memes) and parasitic organisms.

TESTING THE HYPOTHESIS: We predict that next-generation microbiome sequencing of samples obtained from gut or brain tissues of control subjects and subjects with a history of voluntary active participation in certain religious rituals that promote microbial transmission will lead to the discovery of microbes, whose presence has a consistent and positive association with religious behavior. Our hypothesis also predicts a decline of participation in religious rituals in societies with improved sanitation.

If proven true, our hypothesis may provide insights on the origin and pervasiveness of certain religious practices and provide an alternative explanation for recently published positive associations between parasite-stress and religiosity. The discovery of novel microorganisms that affect host behavior may improve our understanding of neurobiology and neurochemistry, while the diversity of such organisms may be of interest to evolutionary biologists and religious scholars.

REVIEWERS: This article was reviewed by Prof. Dan Graur, Dr. Rob Knight and Dr. Eugene Koonin.}, } @article {pmid24988478, year = {2014}, author = {Frost, CL and Pollock, SW and Smith, JE and Hughes, WO}, title = {Wolbachia in the flesh: symbiont intensities in germ-line and somatic tissues challenge the conventional view of Wolbachia transmission routes.}, journal = {PloS one}, volume = {9}, number = {7}, pages = {e95122}, pmid = {24988478}, issn = {1932-6203}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Ants/*microbiology ; Fat Body/microbiology ; Feces/microbiology ; Female ; Germ Cells/microbiology ; Hemolymph/microbiology ; Host-Pathogen Interactions/*physiology ; Ovary/microbiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Symbionts can substantially affect the evolution and ecology of their hosts. The investigation of the tissue-specific distribution of symbionts (tissue tropism) can provide important insight into host-symbiont interactions. Among other things, it can help to discern the importance of specific transmission routes and potential phenotypic effects. The intracellular bacterial symbiont Wolbachia has been described as the greatest ever panzootic, due to the wide array of arthropods that it infects. Being primarily vertically transmitted, it is expected that the transmission of Wolbachia would be enhanced by focusing infection in the reproductive tissues. In social insect hosts, this tropism would logically extend to reproductive rather than sterile castes, since the latter constitute a dead-end for vertically transmission. Here, we show that Wolbachia are not focused on reproductive tissues of eusocial insects, and that non-reproductive tissues of queens and workers of the ant Acromyrmex echinatior, harbour substantial infections. In particular, the comparatively high intensities of Wolbachia in the haemolymph, fat body, and faeces, suggest potential for horizontal transmission via parasitoids and the faecal-oral route, or a role for Wolbachia modulating the immune response of this host. It may be that somatic tissues and castes are not the evolutionary dead-end for Wolbachia that is commonly thought.}, } @article {pmid24984056, year = {2014}, author = {Sicard, M and Bouchon, D and Ceyrac, L and Raimond, R and Thierry, M and Le Clec'h, W and Marcadé, I and Caubet, Y and Grève, P}, title = {Bidirectional cytoplasmic incompatibility caused by Wolbachia in the terrestrial isopod Porcellio dilatatus.}, journal = {Journal of invertebrate pathology}, volume = {121}, number = {}, pages = {28-36}, doi = {10.1016/j.jip.2014.06.007}, pmid = {24984056}, issn = {1096-0805}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm ; DNA, Bacterial/chemistry ; *Host-Pathogen Interactions ; Isopoda/*microbiology ; Phylogeny ; Wolbachia/*physiology ; }, abstract = {In the terrestrial isopod species Porcellio dilatatus, unidirectional Cytoplasmic Incompatibility (CI) between two morphs (P. d. dilatatus and P. d. petiti) caused by a Wolbachia strain (wPet) infecting the morph P. d. petiti has been previously described by experiments initiated four decades ago. Here, we studied another Wolbachia that has been recently detected in a population of the morph P. d. dilatatus. The MLST markers reveal that this Wolbachia is a new strain called wDil distinct from wPet also belonging to the isopod clade of Wolbachia. Quantifications of both Wolbachia strains in the gonads of the two P. dilatatus morphs revealed that all males exhibit similar Wolbachia titers while the titers in females depend on the Wolbachia strain they host. Crossing experiments showed that both wDil and wPet induced partial unidirectional CI with different intensities. Moreover, these two strains induced bidirectional CI when individuals were both infected with one of the two different Wolbachia strains. This way, we demonstrated that P. dilatatus can be infected by two closely related Wolbachia strains (wDil and wPet), that seem to have different modification-rescue systems.}, } @article {pmid24982177, year = {2014}, author = {Nikoh, N and Hosokawa, T and Moriyama, M and Oshima, K and Hattori, M and Fukatsu, T}, title = {Evolutionary origin of insect-Wolbachia nutritional mutualism.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {111}, number = {28}, pages = {10257-10262}, pmid = {24982177}, issn = {1091-6490}, mesh = {Animals ; Base Sequence ; *Bedbugs/metabolism/microbiology ; Biotin/biosynthesis/genetics ; Feeding Behavior/*physiology ; Genes, Bacterial/*physiology ; Genome, Bacterial/*physiology ; Molecular Sequence Data ; Symbiosis/*physiology ; Vitamin B Complex/biosynthesis/genetics ; *Wolbachia/genetics/metabolism ; }, abstract = {Obligate insect-bacterium nutritional mutualism is among the most sophisticated forms of symbiosis, wherein the host and the symbiont are integrated into a coherent biological entity and unable to survive without the partnership. Originally, however, such obligate symbiotic bacteria must have been derived from free-living bacteria. How highly specialized obligate mutualisms have arisen from less specialized associations is of interest. Here we address this evolutionary issue by focusing on an exceptional insect-Wolbachia nutritional mutualism. Although Wolbachia endosymbionts are ubiquitously found in diverse insects and generally regarded as facultative/parasitic associates for their insect hosts, a Wolbachia strain associated with the bedbug Cimex lectularius, designated as wCle, was shown to be essential for host's growth and reproduction via provisioning of B vitamins. We determined the 1,250,060-bp genome of wCle, which was generally similar to the genomes of insect-associated facultative Wolbachia strains, except for the presence of an operon encoding the complete biotin synthetic pathway that was acquired via lateral gene transfer presumably from a coinfecting endosymbiont Cardinium or Rickettsia. Nutritional and physiological experiments, in which wCle-infected and wCle-cured bedbugs of the same genetic background were fed on B-vitamin-manipulated blood meals via an artificial feeding system, demonstrated that wCle certainly synthesizes biotin, and the wCle-provisioned biotin significantly contributes to the host fitness. These findings strongly suggest that acquisition of a single gene cluster consisting of biotin synthesis genes underlies the bedbug-Wolbachia nutritional mutualism, uncovering an evolutionary transition from facultative symbiosis to obligate mutualism facilitated by lateral gene transfer in an endosymbiont lineage.}, } @article {pmid24974378, year = {2014}, author = {Choi, JY and Aquadro, CF}, title = {The coevolutionary period of Wolbachia pipientis infecting Drosophila ananassae and its impact on the evolution of the host germline stem cell regulating genes.}, journal = {Molecular biology and evolution}, volume = {31}, number = {9}, pages = {2457-2471}, pmid = {24974378}, issn = {1537-1719}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; DNA, Bacterial/analysis ; DNA, Mitochondrial/analysis ; Drosophila/classification/*genetics/*microbiology ; Drosophila Proteins/*genetics ; Evolution, Molecular ; Germ Cells/*cytology/metabolism ; Phylogeny ; Selection, Genetic ; Sequence Analysis, DNA/methods ; Stem Cells/metabolism ; Wolbachia/*genetics ; }, abstract = {The endosymbiotic bacteria Wolbachia pipientis is known to infect a wide range of arthropod species yet less is known about the coevolutionary history it has with its hosts. Evidence of highly identical W. pipientis strains in evolutionary divergent hosts suggests horizontal transfer between hosts. For example, Drosophila ananassae is infected with a W. pipientis strain that is nearly identical in sequence to a strain that infects both D. simulans and D. suzukii, suggesting recent horizontal transfer among these three species. However, it is unknown whether the W. pipientis strain had recently invaded all three species or a more complex infectious dynamic underlies the horizontal transfers. Here, we have examined the coevolutionary history of D. ananassae and its resident W. pipientis to infer its period of infection. Phylogenetic analysis of D. ananassae mitochondrial DNA and W. pipientis DNA sequence diversity revealed the current W. pipientis infection is not recent. In addition, we examined the population genetics and molecular evolution of several germline stem cell (GSC) regulating genes of D. ananassae. These studies reveal significant evidence of recent and long-term positive selection at stonewall in D. ananassae, whereas pumillio showed patterns of variation consistent with only recent positive selection. Previous studies had found evidence for adaptive evolution of two key germline differentiation genes, bag of marbles (bam) and benign gonial cell neoplasm (bgcn), in D. melanogaster and D. simulans and proposed that the adaptive evolution at these two genes was driven by arms race between the host GSC and W. pipientis. However, we did not find any statistical departures from a neutral model of evolution for bam and bgcn in D. ananassae despite our new evidence that this species has been infected with W. pipientis for a period longer than the most recent infection in D. melanogaster. In the end, analyzing the GSC regulating genes individually showed two of the seven genes to have evidence of selection. However, combining the data set and fitting a specific population genetic model significant proportion of the nonsynonymous sites across the GSC regulating genes were driven to fixation by positive selection. Clearly the GSC system is under rapid evolution and potentially multiple drivers are causing the rapid evolution.}, } @article {pmid24968670, year = {2014}, author = {Mingchay, P and Sai-Ngam, A and Phumee, A and Bhakdeenuan, P and Lorlertthum, K and Thavara, U and Tawatsin, A and Choochote, W and Siriyasatien, P}, title = {Wolbachia supergroups A and B in natural populations of medically important filth flies (diptera: muscidae, calliphoridae, and sarcophagidae) in Thailand.}, journal = {The Southeast Asian journal of tropical medicine and public health}, volume = {45}, number = {2}, pages = {309-318}, pmid = {24968670}, issn = {0125-1562}, mesh = {Animals ; Diptera/*microbiology ; Female ; Male ; Muscidae/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; Population Density ; Sarcophagidae/*microbiology ; Thailand ; Wolbachia/*isolation & purification/*physiology ; }, abstract = {Filth flies, belonging to suborder Brachycera (Family; Muscidae, Calliphoridae and Sarcophagidae), are a major cause of nuisance and able to transmit pathogens to humans and animals. These insects are distributed worldwide and their populations are increasing especially in sub-tropical and tropical areas. One strategy for controlling insects employs Wolbachia, which is a group of maternally inherited intracellular bacteria, found in many insect species. The bacteria can cause reproductive abnormalities in their hosts, such as cytoplasmic incompatibility, feminization, parthenogenesis, and male lethality. In this study we determined Wolbachia endosymbionts in natural population of medically important flies (42 females and 9 males) from several geographic regions of Thailand. Wolbachia supergroups A or B were detected in 7 of female flies using PCR specific for wsp. Sequence analysis of wsp showed variations between and within the Wolbachia supergroup. Phylogenetics demonstrated that wsp is able to diverge between Wolbachia supergroups A and B. These data should be useful in future Wolbachia-based programs of fly control.}, } @article {pmid24963388, year = {2014}, author = {Augustinos, AA and Asimakopoulou, AK and Moraiti, CA and Mavragani-Tsipidou, P and Papadopoulos, NT and Bourtzis, K}, title = {Microsatellite and Wolbachia analysis in Rhagoletis cerasi natural populations: population structuring and multiple infections.}, journal = {Ecology and evolution}, volume = {4}, number = {10}, pages = {1943-1962}, pmid = {24963388}, issn = {2045-7758}, abstract = {Rhagoletis cerasi (Diptera: Tephritidae) is a major pest of sweet and sour cherries in Europe and parts of Asia. Despite its economic significance, there is a lack of studies on the genetic structure of R. cerasi populations. Elucidating the genetic structure of insects of economic importance is crucial for developing phenological-predictive models and environmental friendly control methods. All natural populations of R. cerasi have been found to harbor the endosymbiont Wolbachia pipientis, which widely affects multiple biological traits contributing to the evolution of its hosts, and has been suggested as a tool for the biological control of insect pests and disease vectors. In the current study, the analysis of 18 R. cerasi populations collected in Greece, Germany, and Russia using 13 microsatellite markers revealed structuring of R. cerasi natural populations, even at close geographic range. We also analyzed the Wolbachia infection status of these populations using 16S rRNA-, MLST- and wsp-based approaches. All 244 individuals screened were positive for Wolbachia. Our results suggest the fixation of the wCer1 strain in Greece while wCer2, wCer4, wCer5, and probably other uncharacterized strains were also detected in multiply infected individuals. The role of Wolbachia and its potential extended phenotypes needs a thorough investigation in R. cerasi. Our data suggest an involvement of this symbiont in the observed restriction in the gene flow in addition to a number of different ecological factors.}, } @article {pmid24958169, year = {2014}, author = {Armstrong, SD and Babayan, SA and Lhermitte-Vallarino, N and Gray, N and Xia, D and Martin, C and Kumar, S and Taylor, DW and Blaxter, ML and Wastling, JM and Makepeace, BL}, title = {Comparative analysis of the secretome from a model filarial nematode (Litomosoides sigmodontis) reveals maximal diversity in gravid female parasites.}, journal = {Molecular & cellular proteomics : MCP}, volume = {13}, number = {10}, pages = {2527-2544}, pmid = {24958169}, issn = {1535-9484}, mesh = {Animals ; Disease Models, Animal ; Female ; Filariasis/blood/*parasitology ; Filarioidea/classification/*growth & development/metabolism ; Gene Expression Regulation, Developmental ; Genetic Variation ; Helminth Proteins/*genetics/metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Proteomics/*methods ; Sex Factors ; }, abstract = {Filarial nematodes (superfamily Filarioidea) are responsible for an annual global health burden of ∼6.3 million disability-adjusted life-years, which represents the greatest single component of morbidity attributable to helminths affecting humans. No vaccine exists for the major filarial diseases, lymphatic filariasis and onchocerciasis; in part because research on protective immunity against filariae has been constrained by the inability of the human-parasitic species to complete their lifecycles in laboratory mice. However, the rodent filaria Litomosoides sigmodontis has become a popular experimental model, as BALB/c mice are fully permissive for its development and reproduction. Here, we provide a comprehensive analysis of excretory-secretory products from L. sigmodontis across five lifecycle stages and identifications of host proteins associated with first-stage larvae (microfilariae) in the blood. Applying intensity-based quantification, we determined the abundance of 302 unique excretory-secretory proteins, of which 64.6% were present in quantifiable amounts only from gravid adult female nematodes. This lifecycle stage, together with immature microfilariae, released four proteins that have not previously been evaluated as vaccine candidates: a predicted 28.5 kDa filaria-specific protein, a zonadhesin and SCO-spondin-like protein, a vitellogenin, and a protein containing six metridin-like ShK toxin domains. Female nematodes also released two proteins derived from the obligate Wolbachia symbiont. Notably, excretory-secretory products from all parasite stages contained several uncharacterized members of the transthyretin-like protein family. Furthermore, biotin labeling revealed that redox proteins and enzymes involved in purinergic signaling were enriched on the adult nematode cuticle. Comparison of the L. sigmodontis adult secretome with that of the human-infective filarial nematode Brugia malayi (reported previously in three independent published studies) identified differences that suggest a considerable underlying diversity of potential immunomodulators. The molecules identified in L. sigmodontis excretory-secretory products show promise not only for vaccination against filarial infections, but for the amelioration of allergy and autoimmune diseases.}, } @article {pmid24948103, year = {2014}, author = {Huber, K and Jansen, S and Leggewie, M and Badusche, M and Schmidt-Chanasit, J and Becker, N and Tannich, E and Becker, SC}, title = {Aedes japonicus japonicus (Diptera: Culicidae) from Germany have vector competence for Japan encephalitis virus but are refractory to infection with West Nile virus.}, journal = {Parasitology research}, volume = {113}, number = {9}, pages = {3195-3199}, pmid = {24948103}, issn = {1432-1955}, mesh = {Aedes/*virology ; Animals ; Coinfection ; Culex/virology ; Encephalitis Virus, Japanese/*physiology ; Feeding Behavior ; Female ; Germany ; Humans ; Insect Vectors/*virology ; West Nile virus/*physiology ; }, abstract = {The interplay between arthropod-borne (arbo) viruses and their vectors is usually complex and often exert unique relationships. Aedes japonicus japonicus (Hulecoeteomyia japonica or Ochlerotatus japonicus japonicus), an invasive mosquito species with laboratory proven vector competence for a number of emerging viruses has been newly introduced to Germany and is currently expanding its range throughout the country. On the other hand, West Nile virus (WNV), an emerging arbovirus originating from Africa, is already circulating in several European countries and might soon be introduced to Germany. Because newly introduced and rapidly expanding vector species pose a potential risk for public health in Germany, we assessed the vectorial capacity of German Ae. j. japonicus populations for WNV and Japanese encephalitis virus (JEV). The results indicate that German Ae. j. japonicus are susceptible for JEV but are refractory to infection with WNV. Of 67 Ae. j. japonicus females challenged by feeding of WNV-containing blood, none had measurable amounts of WNV-RNA (0% infection rate) on day 14 post-infection. In contrast, all females challenged with JEV were positive for JEV-RNA (100% infection rate) on day 14 post-infection. The reason for WNV resistance remains to be determined but is independent from co-infection with other flaviviruses or the presence of endosymbiotic Wolbachia, since we found no evidence for other flavivirus infections within 1,033 tested A. j. japonicus females from the sampling region, nor detectable Wolbachia infection within 30 randomly selected individuals.}, } @article {pmid24945631, year = {2014}, author = {Nag, JK and Shrivastava, N and Chahar, D and Gupta, CL and Bajpai, P and Misra-Bhattacharya, S}, title = {Wolbachia transcription elongation factor "Wol GreA" interacts with α2ββ'σ subunits of RNA polymerase through its dimeric C-terminal domain.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {6}, pages = {e2930}, pmid = {24945631}, issn = {1935-2735}, mesh = {Amino Acid Sequence ; Chromatography, Gel ; Cross-Linking Reagents/metabolism ; DNA-Directed RNA Polymerases/*metabolism ; Fluorometry ; Models, Molecular ; Molecular Docking Simulation ; Molecular Sequence Data ; Phylogeny ; Protein Binding ; Protein Conformation ; *Protein Interaction Domains and Motifs ; *Protein Interaction Mapping ; Transcription Elongation, Genetic ; Transcription Factors/chemistry/*metabolism ; Wolbachia/*enzymology ; }, abstract = {OBJECTIVES: Wolbachia, an endosymbiont of filarial nematode, is considered a promising target for therapy against lymphatic filariasis. Transcription elongation factor GreA is an essential factor that mediates transcriptional transition from abortive initiation to productive elongation by stimulating the escape of RNA polymerase (RNAP) from native prokaryotic promoters. Upon screening of 6257 essential bacterial genes, 57 were suggested as potential future drug targets, and GreA is among these. The current study emphasized the characterization of Wol GreA with its domains.

Biophysical characterization of Wol GreA with its N-terminal domain (NTD) and C-terminal domain (CTD) was performed with fluorimetry, size exclusion chromatography, and chemical cross-linking. Filter trap and far western blotting were used to determine the domain responsible for the interaction with α2ββ'σ subunits of RNAP. Protein-protein docking studies were done to explore residual interaction of RNAP with Wol GreA. The factor and its domains were found to be biochemically active. Size exclusion and chemical cross-linking studies revealed that Wol GreA and CTD exist in a dimeric conformation while NTD subsists in monomeric conformation. Asp120, Val121, Ser122, Lys123, and Ser134 are the residues of CTD through which monomers of Wol GreA interact and shape into a dimeric conformation. Filter trap, far western blotting, and protein-protein docking studies revealed that dimeric CTD of Wol GreA through Lys82, Ser98, Asp104, Ser105, Glu106, Tyr109, Glu116, Asp120, Val121, Ser122, Ser127, Ser129, Lys140, Glu143, Val147, Ser151, Glu153, and Phe163 residues exclusively participates in binding with α2ββ'σ subunits of polymerase.

CONCLUSIONS/SIGNIFICANCE: To the best of our knowledge, this research is the first documentation of the residual mode of action in wolbachial mutualist. Therefore, findings may be crucial to understanding the transcription mechanism of this α-proteobacteria and in deciphering the role of Wol GreA in filarial development.}, } @article {pmid24941309, year = {2014}, author = {Shahab, M and Verma, M and Pathak, M and Mitra, K and Misra-Bhattacharya, S}, title = {Cloning, expression and characterization of UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) from Wolbachia endosymbiont of human lymphatic filarial parasite Brugia malayi.}, journal = {PloS one}, volume = {9}, number = {6}, pages = {e99884}, pmid = {24941309}, issn = {1932-6203}, mesh = {Alkyl and Aryl Transferases/chemistry/genetics/*metabolism ; Amino Acid Sequence ; Animals ; Bacterial Proteins/chemistry/genetics/metabolism ; Brugia malayi/drug effects/growth & development/*microbiology ; Cloning, Molecular ; Female ; Filariasis/*parasitology ; Fosfomycin/pharmacology ; Gene Expression Regulation, Developmental/drug effects ; Humans ; Hydrogen-Ion Concentration ; Kinetics ; Life Cycle Stages ; Lymphoid Tissue/*parasitology/pathology ; Models, Molecular ; Molecular Sequence Data ; Murinae ; Parasites/drug effects/growth & development/*microbiology ; Peptidoglycan/biosynthesis ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Structural Homology, Protein ; *Symbiosis/drug effects ; Temperature ; Wolbachia/drug effects/*enzymology ; }, abstract = {Wolbachia, an endosymbiont of filarial nematode, is considered a promising target for treatment of lymphatic filariasis. Although functional characterization of the Wolbachia peptidoglycan assembly has not been fully explored, the Wolbachia genome provides evidence for coding all of the genes involved in lipid II biosynthesis, a part of peptidoglycan biosynthesis pathway. UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) is one of the lipid II biosynthesis pathway enzymes and it has inevitably been recognized as an antibiotic target. In view of the vital role of MurA in bacterial viability and survival, MurA ortholog from Wolbachia endosymbiont of Brugia malayi (wBm-MurA) was cloned, expressed and purified for further molecular characterization. The enzyme kinetics and inhibition studies were undertaken using fosfomycin. wBm-MurA was found to be expressed in all the major life stages of B. malayi and was immunolocalized in Wolbachia within the microfilariae and female adults by the confocal microscopy. Sequence analysis suggests that the amino acids crucial for enzymatic activity are conserved. The purified wBm-MurA was shown to possess the EPSP synthase (3-phosphoshikimate 1-carboxyvinyltransferase) like activity at a broad pH range with optimal activity at pH 7.5 and 37°C temperature. The apparent affinity constant (Km) for the substrate UDP-N-acetylglucosamine was found to be 0.03149 mM and for phosphoenolpyruvate 0.009198 mM. The relative enzymatic activity was inhibited ∼2 fold in presence of fosfomycin. Superimposition of the wBm-MurA homology model with the structural model of Haemophilus influenzae (Hi-MurA) suggests binding of fosfomycin at the same active site. The findings suggest wBm-MurA to be a putative antifilarial drug target for screening of novel compounds.}, } @article {pmid24929215, year = {2014}, author = {Nag, JK and Shrivastava, N and Tiwari, M and Gupta, Cl and Bajpai, P and Chahar, D and Misra-Bhattacharya, S}, title = {Wolbachia translation initiation factor-1 is copiously expressed by the adult, microfilariae and infective larvae of Brugia malayi and competitively inhibited by tetracycline.}, journal = {Acta tropica}, volume = {138}, number = {}, pages = {51-59}, doi = {10.1016/j.actatropica.2014.04.033}, pmid = {24929215}, issn = {1873-6254}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/*microbiology ; Female ; Gene Expression Profiling ; Immunoblotting ; Male ; Mice, Inbred BALB C ; Microscopy, Confocal ; Models, Molecular ; Prokaryotic Initiation Factor-1/*biosynthesis/chemistry/isolation & purification ; Protein Binding/drug effects ; Protein Biosynthesis/*drug effects ; Protein Conformation ; Protein Folding ; Ribosomal Proteins/metabolism ; Ribosome Subunits, Small, Bacterial/metabolism ; Tetracycline/*pharmacology ; Wolbachia/*drug effects/*genetics ; }, abstract = {The intracellular alphaproteobacteria, Wolbachia, is considered to be a future antimacrofilarial drug target as it is obligatory for filarial endurance. Characterizing wolbachial proteins is necessary to understand wolbachial mechanisms and also for discovering new drug entities. Translation initiation factor-1 (Tl IF-1) is an indispensable prokaryotic factor concerned with bacterial viability. This factor is prioritized as one of the most potent antibacterial drug target. To investigate its role in filarial biology, recombinant Wol Tl IF-1 was purified on metal ion column. The factor was found folded in its monomeric native conformation, and contained a buried fluorophore. Molecular modeling revealed that the factor belonged to the Oligomer Binding family, and consisted of the highly conserved S1 domain with 81.6% of the amino acids occupying the allowed regions in Ramachandran plot. In addition, Wol Tl IF-1 exhibited selective binding to the 30S ribosomal subunit, which declined progressively with tetracycline addition. Tetracycline perturbs interaction of Thr18 and Asn32 of the factor with ribosomal protein S4. The factor was immune-localized in adult, microfilariae (Mf) and infective larvae (L3) of Brugia malayi by immunoblotting. High expression was also observed in Wolbachia within B. malayi Mf, L3 and female adult parasite along the gravid uteri by the confocal microscopy. Therefore, Wol Tl IF-1 appears to be an essential Wolbachia factor whose inhibition leads to extensive cell apoptosis and premature killing of adult worms, validating the antifilarial potential of the factor.}, } @article {pmid24928884, year = {2014}, author = {Yun, JH and Roh, SW and Whon, TW and Jung, MJ and Kim, MS and Park, DS and Yoon, C and Nam, YD and Kim, YJ and Choi, JH and Kim, JY and Shin, NR and Kim, SH and Lee, WJ and Bae, JW}, title = {Insect gut bacterial diversity determined by environmental habitat, diet, developmental stage, and phylogeny of host.}, journal = {Applied and environmental microbiology}, volume = {80}, number = {17}, pages = {5254-5264}, pmid = {24928884}, issn = {1098-5336}, mesh = {Animals ; *Biodiversity ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Diet ; Ecosystem ; Gastrointestinal Tract/microbiology ; Insecta/*microbiology/physiology ; Life Cycle Stages ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {Insects are the most abundant animals on Earth, and the microbiota within their guts play important roles by engaging in beneficial and pathological interactions with these hosts. In this study, we comprehensively characterized insect-associated gut bacteria of 305 individuals belonging to 218 species in 21 taxonomic orders, using 454 pyrosequencing of 16S rRNA genes. In total, 174,374 sequence reads were obtained, identifying 9,301 bacterial operational taxonomic units (OTUs) at the 3% distance level from all samples, with an average of 84.3 (± 97.7) OTUs per sample. The insect gut microbiota were dominated by Proteobacteria (62.1% of the total reads, including 14.1% Wolbachia sequences) and Firmicutes (20.7%). Significant differences were found in the relative abundances of anaerobes in insects and were classified according to the criteria of host environmental habitat, diet, developmental stage, and phylogeny. Gut bacterial diversity was significantly higher in omnivorous insects than in stenophagous (carnivorous and herbivorous) insects. This insect-order-spanning investigation of the gut microbiota provides insights into the relationships between insects and their gut bacterial communities.}, } @article {pmid24926801, year = {2014}, author = {da Rocha Fernandes, M and Martins, R and Pessoa Costa, E and Pacidônio, EC and Araujo de Abreu, L and da Silva Vaz, I and Moreira, LA and da Fonseca, RN and Logullo, C}, title = {The modulation of the symbiont/host interaction between Wolbachia pipientis and Aedes fluviatilis embryos by glycogen metabolism.}, journal = {PloS one}, volume = {9}, number = {6}, pages = {e98966}, pmid = {24926801}, issn = {1932-6203}, mesh = {Aedes/*embryology/metabolism/*microbiology ; Animals ; Embryo, Nonmammalian/microbiology ; Embryonic Development/physiology ; Female ; Glucose-6-Phosphate/metabolism ; Glycogen/*metabolism ; Glycogen Synthase Kinase 3/genetics ; Host-Parasite Interactions/*physiology ; Male ; Phylogeny ; Symbiosis/*physiology ; Wolbachia/metabolism/*physiology ; }, abstract = {Wolbachia pipientis, a maternally transmitted bacterium that colonizes arthropods, may affect the general aspects of insect physiology, particularly reproduction. Wolbachia is a natural endosymbiont of Aedes fluviatilis, whose effects in embryogenesis and reproduction have not been addressed so far. In this context, we investigated the correlation between glucose metabolism and morphological alterations during A. fluviatilis embryo development in Wolbachia-positive (W+) and Wolbachia-negative (W-) mosquito strains. While both strains do not display significant morphological and larval hatching differences, larger differences were observed in hexokinase activity and glycogen contents during early and mid-stages of embryogenesis, respectively. To investigate if glycogen would be required for parasite-host interaction, we reduced Glycogen Synthase Kinase-3 (GSK-3) levels in adult females and their eggs by RNAi. GSK-3 knock-down leads to embryonic lethality, lower levels of glycogen and total protein and Wolbachia reduction. Therefore, our results suggest that the relationship between A. fluviatilis and Wolbachia may be modulated by glycogen metabolism.}, } @article {pmid24912954, year = {2014}, author = {Landum, M and Ferreira, CC and Calado, M and Alho, AM and Maurício, IL and Meireles, JS and de Carvalho, LM and Cunha, C and Belo, S}, title = {Detection of Wolbachia in Dirofilaria infected dogs in Portugal.}, journal = {Veterinary parasitology}, volume = {204}, number = {3-4}, pages = {407-410}, doi = {10.1016/j.vetpar.2014.05.027}, pmid = {24912954}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Dirofilaria immitis/*immunology/isolation & purification ; Dirofilariasis/*parasitology/prevention & control ; Dog Diseases/*parasitology/prevention & control ; Dogs ; Doxycycline/*therapeutic use ; Female ; Male ; Microfilariae ; Portugal/epidemiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia pipiens, an intracellular endosymbiont bacteria of filarial nematodes, has been implicated in the pathogenesis of filarial diseases, in particular in heavy Dirofilaria spp. infections. Antibiotic therapy (doxycycline) against Wolbachia has been proven to be suitable adjunct therapy, prior to adulticide treatment of canine dirofilariosis. Despite its importance, investigation on the Wolbachia/Dirofilaria complex in Portugal had not been undertaken so far. This study reports the first detection of Wolbachia in Dirofilaria spp. infected dogs in the context of an ongoing epidemiological survey in central-south regions in the country. Wolbachia DNA was detected by PCR in 52.6% (20/38) of canine blood samples positive for Dirofilaria immitis based on parasitological (Knott's and Acid Phosphatase) and serological (Witness(®)Dirofilaria) methods. No Wolbachia DNA could be detected in samples from dogs with occult infections (parasite negative but antigen positive). The lack of Wolbachia detection in some microfilaremic dogs was somewhat unexpected and needs to be elucidated in further studies, as the presence or absence of these bacteria in association with microfilaria is of importance for veterinarians in the management and control of canine dirofilariosis.}, } @article {pmid24911519, year = {2014}, author = {Chrostek, E and Marialva, MS and Yamada, R and O'Neill, SL and Teixeira, L}, title = {High anti-viral protection without immune upregulation after interspecies Wolbachia transfer.}, journal = {PloS one}, volume = {9}, number = {6}, pages = {e99025}, pmid = {24911519}, issn = {1932-6203}, support = {//Wellcome Trust/United Kingdom ; 094664/Z/10/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila melanogaster/*immunology/microbiology/*virology ; Evolution, Molecular ; Longevity ; Phenotype ; Polymorphism, Genetic ; Symbiosis ; Wolbachia/*immunology/*virology ; }, abstract = {Wolbachia, endosymbionts that reside naturally in up to 40-70% of all insect species, are some of the most prevalent intracellular bacteria. Both Wolbachia wAu, naturally associated with Drosophila simulans, and wMel, native to Drosophila melanogaster, have been previously described to protect their hosts against viral infections. wMel transferred to D. simulans was also shown to have a strong antiviral effect. Here we directly compare one of the most protective wMel variants and wAu in D. melanogaster in the same host genetic background. We conclude that wAu protects better against viral infections, it grows exponentially and significantly shortens the lifespan of D. melanogaster. However, there is no difference between wMel and wAu in the expression of selected antimicrobial peptides. Therefore, neither the difference in anti-viral effect nor the life-shortening could be attributed to the immune stimulation by exogenous Wolbachia. Overall, we prove that stable transinfection with a highly protective Wolbachia is not necessarily associated with general immune activation.}, } @article {pmid24909063, year = {2014}, author = {Tamarozzi, F and Wright, HL and Johnston, KL and Edwards, SW and Turner, JD and Taylor, MJ}, title = {Human filarial Wolbachia lipopeptide directly activates human neutrophils in vitro.}, journal = {Parasite immunology}, volume = {36}, number = {10}, pages = {494-502}, pmid = {24909063}, issn = {1365-3024}, mesh = {Animals ; Apoptosis ; Chemotaxis ; Humans ; Interleukin-8/immunology ; Lipopeptides/*immunology ; Neutrophils/*immunology/pathology ; Onchocerca volvulus/*microbiology ; Onchocerciasis, Ocular/*immunology/parasitology ; Respiratory Burst ; Wolbachia/*immunology ; }, abstract = {The host inflammatory response to the Onchocerca volvulus endosymbiont, Wolbachia, is a major contributing factor in the development of chronic pathology in humans (onchocerciasis/river blindness). Recently, the toll-like pattern recognition receptor motif of the major inflammatory ligands of filarial Wolbachia, membrane-associated diacylated lipoproteins, was functionally defined in murine models of pathology, including mediation of neutrophil recruitment to the cornea. However, the extent to which human neutrophils can be activated in response to this Wolbachia pattern recognition motif is not known. Therefore, the responses of purified peripheral blood human neutrophils to a synthetic N-terminal diacylated lipopeptide (WoLP) of filarial Wolbachia peptidoglycan-associated lipoprotein (PAL) were characterized. WoLP exposure led to a dose-dependent activation of healthy, human neutrophils that included gross morphological alterations and modulation of surface expressed integrins involved in tethering, rolling and extravasation. WoLP exposure induced chemotaxis but not chemokinesis of neutrophils, and secretion of the major neutrophil chemokine, interleukin 8. WoLP also induced and primed the respiratory burst, and enhanced neutrophil survival by delay of apoptosis. These results indicate that the major inflammatory motif of filarial Wolbachia lipoproteins directly activates human neutrophils in vitro and promotes a molecular pathway by which human neutrophils are recruited to sites of Onchocerca parasitism.}, } @article {pmid24905191, year = {2014}, author = {Baldini, F and Segata, N and Pompon, J and Marcenac, P and Shaw, WR and Dabiré, RK and Diabaté, A and Levashina, EA and Catteruccia, F}, title = {Evidence of natural Wolbachia infections in field populations of Anopheles gambiae.}, journal = {Nature communications}, volume = {5}, number = {}, pages = {3985}, pmid = {24905191}, issn = {2041-1723}, support = {093553/WT_/Wellcome Trust/United Kingdom ; 260897/ERC_/European Research Council/International ; R01 AI104956/AI/NIAID NIH HHS/United States ; NIH 1R01AI104956-01A1/AI/NIAID NIH HHS/United States ; BB/I002898/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Anopheles/*microbiology ; Burkina Faso ; Female ; Gram-Negative Bacterial Infections/microbiology/transmission ; Male ; Mosquito Control ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia are maternally transmitted intracellular bacteria that invade insect populations by manipulating their reproduction and immunity and thus limiting the spread of numerous human pathogens. Experimental Wolbachia infections can reduce Plasmodium numbers in Anopheles mosquitoes in the laboratory, however, natural Wolbachia infections in field anophelines have never been reported. Here we show evidence of Wolbachia infections in Anopheles gambiae in Burkina Faso, West Africa. Sequencing of the 16S rRNA gene identified Wolbachia sequences in both female and male germlines across two seasons, and determined that these sequences are vertically transmitted from mother to offspring. Whole-genome sequencing of positive samples suggests that the genetic material identified in An. gambiae belongs to a novel Wolbachia strain, related to but distinct from strains infecting other arthropods. The evidence of Wolbachia infections in natural Anopheles populations promotes further investigations on the possible use of natural Wolbachia-Anopheles associations to limit malaria transmission.}, } @article {pmid24903665, year = {2014}, author = {Gehringer, C and Kreidenweiss, A and Flamen, A and Antony, JS and Grobusch, MP and Bélard, S}, title = {Molecular evidence of Wolbachia endosymbiosis in Mansonella perstans in Gabon, Central Africa.}, journal = {The Journal of infectious diseases}, volume = {210}, number = {10}, pages = {1633-1638}, doi = {10.1093/infdis/jiu320}, pmid = {24903665}, issn = {1537-6613}, mesh = {Adult ; Animals ; Bacterial Proteins/genetics ; Child ; Child, Preschool ; Cytoskeletal Proteins/genetics ; Female ; Gabon ; Humans ; Male ; Mansonella/*microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/genetics/*isolation & purification/*physiology ; }, abstract = {The discovery of obligatory intracellular bacteria of the genus Wolbachia in filariae infecting humans led to the use of antibiotics as a potent treatment option. Mansonella perstans is the cause of the second most prevalent filariasis in Gabon, but so far reports on the presence of Wolbachia in this nematode have been inconsistent. We report on the presence of Wolbachia in M. perstans in patients from Gabon, which we identified using polymerase chain reaction (PCR) with primer sets specific for 16S rDNA and ftsZ. Sequence analysis revealed a single consensus sequence, which could be phylogenetically assigned to Wolbachia of the supergroup F. Wolbachia could only be identified in 5 of 14 or 7 of 14 cases, depending on the investigated gene; detection of Wolbachia was associated with higher-level filaremia. Before generalizing the use of antibiotics for mansonellosis, further clarification of the obligatory nature of the endosymbiosis in this nematode is needed.}, } @article {pmid24901418, year = {2014}, author = {Koutsovoulos, G and Makepeace, B and Tanya, VN and Blaxter, M}, title = {Palaeosymbiosis revealed by genomic fossils of Wolbachia in a strongyloidean nematode.}, journal = {PLoS genetics}, volume = {10}, number = {6}, pages = {e1004397}, pmid = {24901418}, issn = {1553-7404}, support = {095831//Wellcome Trust/United Kingdom ; /BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Base Sequence ; Cattle ; Cattle Diseases/microbiology/parasitology ; DNA, Bacterial/*genetics ; DNA, Protozoan/*genetics ; Dictyocaulus/*genetics/*microbiology ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {Wolbachia are common endosymbionts of terrestrial arthropods, and are also found in nematodes: the animal-parasitic filaria, and the plant-parasite Radopholus similis. Lateral transfer of Wolbachia DNA to the host genome is common. We generated a draft genome sequence for the strongyloidean nematode parasite Dictyocaulus viviparus, the cattle lungworm. In the assembly, we identified nearly 1 Mb of sequence with similarity to Wolbachia. The fragments were unlikely to derive from a live Wolbachia infection: most were short, and the genes were disabled through inactivating mutations. Many fragments were co-assembled with definitively nematode-derived sequence. We found limited evidence of expression of the Wolbachia-derived genes. The D. viviparus Wolbachia genes were most similar to filarial strains and strains from the host-promiscuous clade F. We conclude that D. viviparus was infected by Wolbachia in the past, and that clade F-like symbionts may have been the source of filarial Wolbachia infections.}, } @article {pmid24891730, year = {2014}, author = {Salunkhe, RC and Narkhede, KP and Shouche, YS}, title = {Distribution and evolutionary impact of wolbachia on butterfly hosts.}, journal = {Indian journal of microbiology}, volume = {54}, number = {3}, pages = {249-254}, pmid = {24891730}, issn = {0046-8991}, abstract = {Wolbachia are maternally inherited endosymbiotic alpha-proteobacteria found in terrestrial arthropods and filarial nematodes. They are transmitted vertically through host cytoplasm and alter host biology by inducing various reproductive alterations, like feminization, parthenogenesis, male killing (MK) and cytoplasmic incompatibility. In butterflies, some effects especially MK and sperm-egg incompatibility are well established. All these effects skew the sex ratio towards female and subsequently favor the vertical transmission of Wolbachia. Some of the insects are also infected with multiple Wolbachia strains which may results in some complex phenomenon. In the present review the potential of Wolbachia for promoting evolutionary changes in its hosts with emphasis on recent advances in interactions of butterfly-Wolbachia is discussed. In addition to this, strain diversity of Wolbachia and its effects on various butterfly hosts are also highlighted.}, } @article {pmid24885509, year = {2014}, author = {Gonçalves, Dda S and Cassimiro, AP and de Oliveira, CD and Rodrigues, NB and Moreira, LA}, title = {Wolbachia detection in insects through LAMP: loop mediated isothermal amplification.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {228}, pmid = {24885509}, issn = {1756-3305}, mesh = {Animals ; Biological Control Agents ; Insecta/*microbiology ; Nucleic Acid Amplification Techniques/economics/*methods ; Sensitivity and Specificity ; Time Factors ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: The bacterium Wolbachia is a promising agent for the biological control of vector-borne diseases as some strains have the ability to block the transmission of key human disease-causing pathogens. Fast, accurate and inexpensive methods of differentiating between infected and uninfected insects will be of critical importance as field-based trials of Wolbachia-based bio-control become increasingly common.

FINDINGS: We have developed a specific and sensitive method of detecting Wolbachia based on the isothermal DNA amplification. This technique can be performed in an ordinary heat block without the need for gel-based visualisation, and is effective for a wide variety of insect hosts.

CONCLUSION: Here we present the development of a rapid, highly sensitive and inexpensive method to detect Wolbachia in a variety of insect hosts, including key mosquito disease vectors.}, } @article {pmid24885505, year = {2014}, author = {Schneider, DI and Klasson, L and Lind, AE and Miller, WJ}, title = {More than fishing in the dark: PCR of a dispersed sequence produces simple but ultrasensitive Wolbachia detection.}, journal = {BMC microbiology}, volume = {14}, number = {}, pages = {121}, pmid = {24885505}, issn = {1471-2180}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; DNA, Bacterial/genetics/*isolation & purification ; Drosophila/microbiology/physiology ; Hymenoptera/microbiology/physiology ; *Interspersed Repetitive Sequences ; Polymerase Chain Reaction/*methods ; *Symbiosis ; Tsetse Flies/microbiology/physiology ; Wolbachia/*isolation & purification/physiology ; }, abstract = {BACKGROUND: Detecting intracellular bacterial symbionts can be challenging when they persist at very low densities. Wolbachia, a widespread bacterial endosymbiont of invertebrates, is particularly challenging. Although it persists at high titers in many species, in others its densities are far below the detection limit of classic end-point Polymerase Chain Reaction (PCR). These low-titer infections can be reliably detected by combining PCR with DNA hybridization, but less elaborate strategies based on end-point PCR alone have proven less sensitive or less general.

RESULTS: We introduce a multicopy PCR target that allows fast and reliable detection of A-supergroup Wolbachia--even at low infection titers--with standard end-point PCR. The target is a multicopy motif (designated ARM: A-supergroup repeat motif) discovered in the genome of wMel (the Wolbachia in Drosophila melanogaster). ARM is found in at least seven other Wolbachia A-supergroup strains infecting various Drosophila, the wasp Muscidifurax and the tsetse fly Glossina. We demonstrate that end-point PCR targeting ARM can reliably detect both high- and low-titer Wolbachia infections in Drosophila, Glossina and interspecific hybrids.

CONCLUSIONS: Simple end-point PCR of ARM facilitates detection of low-titer Wolbachia A-supergroup infections. Detecting these infections previously required more elaborate procedures. Our ARM target seems to be a general feature of Wolbachia A-supergroup genomes, unlike other multicopy markers such as insertion sequences (IS).}, } @article {pmid24860790, year = {2014}, author = {Minard, G and Tran, FH and Dubost, A and Tran-Van, V and Mavingui, P and Moro, CV}, title = {Pyrosequencing 16S rRNA genes of bacteria associated with wild tiger mosquito Aedes albopictus: a pilot study.}, journal = {Frontiers in cellular and infection microbiology}, volume = {4}, number = {}, pages = {59}, pmid = {24860790}, issn = {2235-2988}, mesh = {Aedes/*microbiology ; Animals ; Bacteria/classification/*genetics ; Biodiversity ; Computational Biology ; Female ; Microbiota ; Pilot Projects ; RNA, Bacterial ; *RNA, Ribosomal, 16S ; Sequence Analysis, DNA ; }, abstract = {The Asian tiger mosquito Aedes (Stegomya) albopictus is an invasive species that has spread across the world in the last two decades, showing a great capacity to adapt to contrasting climates and environments. While demonstrated in many insects, the contribution of bacterial symbionts in Aedes ecology is a challenging aspect that needs to be investigated. Also some bacterial species have already been identified in Ae. albopictus using classical methods, but a more accurate survey of mosquito-associated bacterial diversity is needed to decipher the potential biological functions of bacterial symbionts in mediating or constraining insect adaptation. We surveyed the bacteria associated with field populations of Ae. albopictus from Madagascar by pyrosequencing 16S rRNA gene amplicons. Different aspects of amplicon preparation and sequencing depth were tested to optimize the breadth of bacterial diversity identified. The results revealed that all mosquitoes collected from different sites have a bacterial microbiota dominated by a single taxon, Wolbachia pipientis, which accounted for about 99% of all 92,615 sequences obtained. As Ae. albopictus is known to harbor two Wolbachia strains (wAlbA and wAlbB), a quantitative PCR was used to estimate the relative densities, (i.e., the bacteria-to-host gene ratios) of each strains in individual mosquitoes. Relative densities were between 6.25 × 10(0.01) and 5.47 × 10(0.1) for wAlbA and between 2.03 × 10(0.1) and 1.4 × 10(1) for wAlbB. Apart from Wolbachia, a total of 31 bacterial taxa were identified at the genus level using different method variations. Diversity index values were low and probably underestimated the true diversity due to the high abundance of Wolbachia sequences vastly outnumbering sequences from other taxa. Further studies should implement alternative strategies to specifically discard from analysis any sequences from Wolbachia, the dominant endosymbiotic bacterium in Ae. albopictus from this area.}, } @article {pmid24853391, year = {2014}, author = {McNaughton, D and Duong, TT}, title = {Designing a community engagement framework for a new dengue control method: a case study from central Vietnam.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {5}, pages = {e2794}, pmid = {24853391}, issn = {1935-2735}, mesh = {Adolescent ; Adult ; Aedes/microbiology/virology ; Animals ; *Community-Institutional Relations ; Dengue/*prevention & control/transmission ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Information Dissemination/*methods ; Interviews as Topic ; Male ; Middle Aged ; Mosquito Control/*methods/organization & administration ; Pest Control, Biological/*methods/organization & administration ; *Public Opinion ; Vietnam ; Wolbachia ; Young Adult ; }, abstract = {BACKGROUND: The Wolbachia strategy aims to manipulate mosquito populations to make them incapable of transmitting dengue viruses between people. To test its efficacy, this strategy requires field trials. Public consultation and engagement are recognized as critical to the future success of these programs, but questions remain regarding how to proceed. This paper reports on a case study where social research was used to design a community engagement framework for a new dengue control method, at a potential release site in central Vietnam.

The approach described here, draws on an anthropological methodology and uses both qualitative and quantitative methods to design an engagement framework tailored to the concerns, expectations, and socio-political setting of a potential trial release site for Wolbachia-infected Aedes aegypti mosquitoes. The process, research activities, key findings and how these were responded to are described. Safety of the method to humans and the environment was the most common and significant concern, followed by efficacy and impact on local lives. Residents expected to be fully informed and engaged about the science, the project, its safety, the release and who would be responsible should something go wrong. They desired a level of engagement that included regular updates and authorization from government and at least one member of every household at the release site.

CONCLUSIONS/SIGNIFICANCE: Results demonstrate that social research can provide important and reliable insights into public concerns and expectations at a potential release site, as well as guidance on how these might be addressed. Findings support the argument that using research to develop more targeted, engagement frameworks can lead to more sensitive, thorough, culturally comprehensible and therefore ethical consultation processes. This approach has now been used successfully to seek public input and eventually support for releases Wolbachia-infected mosquitoes, in two different international settings--Australia and Vietnam.}, } @article {pmid24850893, year = {2014}, author = {Vanthournout, B and Deswarte, K and Hammad, H and Bilde, T and Lambrecht, B and Hendrickx, F}, title = {Flow cytometric sexing of spider sperm reveals an equal sperm production ratio in a female-biased species.}, journal = {Biology letters}, volume = {10}, number = {5}, pages = {20140159}, pmid = {24850893}, issn = {1744-957X}, support = {282163/ERC_/European Research Council/International ; }, mesh = {Animals ; Female ; Flow Cytometry ; Male ; *Sex Ratio ; Spermatogenesis ; *Spermatozoa ; *Spiders ; }, abstract = {Producing equal amounts of male and female offspring has long been considered an evolutionarily stable strategy. Nevertheless, exceptions to this general rule (i.e. male and female biases) are documented in many taxa, making sex allocation an important domain in current evolutionary biology research. Pinpointing the underlying mechanism of sex ratio bias is challenging owing to the multitude of potential sex ratio-biasing factors. In the dwarf spider, Oedothorax gibbosus, infection with the bacterial endosymbiont Wolbachia results in a female bias. However, pedigree analysis reveals that other factors influence sex ratio variation. In this paper, we investigate whether this additional variation can be explained by the unequal production of male- and female-determining sperm cells during sperm production. Using flow cytometry, we show that males produce equal amounts of male- and female-determining sperm cells; thus bias in sperm production does not contribute to the sex ratio bias observed in this species. This demonstrates that other factors such as parental genes suppressing endosymbiont effects and cryptic female choice might play a role in sex allocation in this species.}, } @article {pmid24831608, year = {2014}, author = {Liu, HY and Wang, YK and Zhi, CC and Xiao, JH and Huang, DW}, title = {A novel approach to eliminate Wolbachia infections in Nasonia vitripennis revealed different antibiotic resistance between two bacterial strains.}, journal = {FEMS microbiology letters}, volume = {355}, number = {2}, pages = {163-169}, doi = {10.1111/1574-6968.12471}, pmid = {24831608}, issn = {1574-6968}, mesh = {Animals ; DNA, Bacterial/genetics ; *Drug Resistance, Multiple, Bacterial ; Female ; Male ; Real-Time Polymerase Chain Reaction ; Reproduction ; Rickettsiaceae Infections/*prevention & control ; Rifampin/pharmacology ; Sequence Analysis, DNA ; Species Specificity ; Wasps/*microbiology ; Wolbachia/classification/*drug effects ; }, abstract = {Wolbachia are widespread in insects and can manipulate host reproduction. Nasonia vitripennis is a widely studied organism with a very high prevalence of Wolbachia infection. To study the effect of Wolbachia infection in Nasonia spp., it is important to obtain noninfected individuals by artificial methods. Current methods that employ sugar water-containing antibiotics can successfully eliminate Wolbachia from the parasitic wasps; however, treatment of at least three generations is required. Here, we describe a novel, feasible, and effective approach to eliminate Wolbachia from N. vitripennis by feeding fly pupae continuously offering antibiotics to Nasonia populations, which shortened the time to eliminate the pathogens to two generations. Additionally, the Wolbachia Uni and CauB strains have obviously different rifampicin-resistance abilities, which is a previously unknown phenomenon.}, } @article {pmid24824176, year = {2015}, author = {Vieira, L and Silvestre-Ferreira, AC and Fontes-Sousa, AP and Balreira, AC and Morchón, R and Carretón, E and Vilhena, H and Simón, F and Montoya-Alonso, JA}, title = {Seroprevalence of heartworm (Dirofilaria immitis) in feline and canine hosts from central and northern Portugal.}, journal = {Journal of helminthology}, volume = {89}, number = {5}, pages = {625-629}, doi = {10.1017/S0022149X14000352}, pmid = {24824176}, issn = {1475-2697}, mesh = {Animals ; Antigens, Helminth/blood ; Cat Diseases/*blood/epidemiology/parasitology ; Cats ; Dirofilaria immitis/immunology/*isolation & purification ; Dirofilariasis/*blood/epidemiology/parasitology ; Dog Diseases/*blood/epidemiology/parasitology ; Dogs ; Female ; Male ; Portugal/epidemiology ; Prevalence ; Seroepidemiologic Studies ; }, abstract = {Dirofilaria immitis is endemic in Portugal. Several studies have reported the presence of canine heartworm disease, although no previous studies on feline infections have been published. The aim of this study was to determine the prevalence of D. immitis in cats and dogs from central and northern Portugal. Blood samples from 434 cats were tested for circulating anti-D. immitis and anti-Wolbachia antibodies. Furthermore, 386 dogs were tested for circulating D. immitis antigens. Overall feline seroprevalence was 15%, while canine prevalence was 2.1%. The highest feline seroprevalences of 18.7% and 17.6% were found in Aveiro and Viseu, respectively, while the highest canine prevalences of 8.8% and 6.8% were found in Coimbra and Aveiro, respectively. Cats and dogs showing respiratory signs presented higher prevalences of 24.4% and 17%, respectively, while 50% of cats with gastrointestinal signs were seropositive. The present study confirms the seropositivity of D. immitis in the feline population in central and northern Portugal, and suggests the importance of including heartworm disease in the list of differential diagnoses of cats and dogs showing clinical signs compatible with the disease.}, } @article {pmid24821918, year = {2014}, author = {Burt, A}, title = {Heritable strategies for controlling insect vectors of disease.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {369}, number = {1645}, pages = {20130432}, pmid = {24821918}, issn = {1471-2970}, mesh = {Animals ; *Biological Control Agents ; Culicidae/genetics/*microbiology/pathogenicity ; Cytoplasm/microbiology ; Dengue/prevention & control ; Humans ; Insect Vectors/genetics/*microbiology ; Longevity ; Malaria/prevention & control ; Mosquito Control/*methods/*trends ; Wolbachia/*genetics ; }, abstract = {Mosquito-borne diseases are causing a substantial burden of mortality, morbidity and economic loss in many parts of the world, despite current control efforts, and new complementary approaches to controlling these diseases are needed. One promising class of new interventions under development involves the heritable modification of the mosquito by insertion of novel genes into the nucleus or of Wolbachia endosymbionts into the cytoplasm. Once released into a target population, these modifications can act to reduce one or more components of the mosquito population's vectorial capacity (e.g. the number of female mosquitoes, their longevity or their ability to support development and transmission of the pathogen). Some of the modifications under development are designed to be self-limiting, in that they will tend to disappear over time in the absence of recurrent releases (and hence are similar to the sterile insect technique, SIT), whereas other modifications are designed to be self-sustaining, spreading through populations even after releases stop (and hence are similar to traditional biological control). Several successful field trials have now been performed with Aedes mosquitoes, and such trials are helping to define the appropriate developmental pathway for this new class of intervention.}, } @article {pmid24820578, year = {2014}, author = {Hughes, GL and Samuels, SK and Shaikh, K and Rasgon, JL and Vardo-Zalik, AM}, title = {Discrimination of the Plasmodium mexicanum vectors Lutzomyia stewarti and Lutzomyia vexator by a PCR-RFLP assay and Wolbachia infection.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {39}, number = {1}, pages = {224-227}, pmid = {24820578}, issn = {1948-7134}, support = {R21 AI111175/AI/NIAID NIH HHS/United States ; R21 AI070178/AI/NIAID NIH HHS/United States ; R01 AI067371/AI/NIAID NIH HHS/United States ; R01AI067371/AI/NIAID NIH HHS/United States ; R21AI111175/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Insect Vectors/classification/genetics/microbiology/parasitology ; Plasmodium/*pathogenicity ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Psychodidae/*classification/genetics/*microbiology/parasitology ; Wolbachia/*pathogenicity ; }, } @article {pmid24819964, year = {2014}, author = {Zabal-Aguirre, M and Arroyo, F and García-Hurtado, J and de la Torre, J and Hewitt, GM and Bella, JL}, title = {Wolbachia effects in natural populations of Chorthippus parallelus from the Pyrenean hybrid zone.}, journal = {Journal of evolutionary biology}, volume = {27}, number = {6}, pages = {1136-1148}, doi = {10.1111/jeb.12389}, pmid = {24819964}, issn = {1420-9101}, mesh = {Animals ; Female ; Fertility ; Grasshoppers/*microbiology ; *Host-Pathogen Interactions ; Hybridization, Genetic ; Male ; Wolbachia/*physiology ; }, abstract = {We evaluate for the first time the effect of Wolbachia infection, involving two different supergroups, on the structure and dynamics of the hybrid zone between two subspecies of Chorthippus parallelus (Orthoptera) in the Pyrenees. Wolbachia infection showed no effects on female fecundity or a slight increment in females infected by F supergroup, although in the last case it has to be well established. Cytoplasmic incompatibility (CI) is confirmed in crosses carried out in the field between individuals from a natural hybrid population. This CI, registered as the relative reduction in embryo production (sh), was of sh = 0.355 and sh = 0.286 in unidirectional crosses involving B and F supergroups, respectively. CI also occurred in bidirectional crosses (sh = 0.147) but with a weaker intensity. The transmission rates of the two Wolbachia strains (B and F) were estimated by the optimization of a theoretical model to reach the infection frequencies observed in certain population. To fit this scenario, both supergroups should present transmission rates close to 1. Further, we have simulated the infection dynamics, and hence, the capacity of Wolbachia to structure the population of the host insects and to affect to reproduction and genetic introgression in the hybrid zone. This represents a first example of the influence of Wolbachia in an insect natural hybrid zone.}, } @article {pmid24813786, year = {2014}, author = {Kramer, L and Genchi, C}, title = {Where are we with Wolbachia and doxycycline: an in-depth review of the current state of our knowledge.}, journal = {Veterinary parasitology}, volume = {206}, number = {1-2}, pages = {1-4}, doi = {10.1016/j.vetpar.2014.03.028}, pmid = {24813786}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology/therapeutic use ; Antinematodal Agents/pharmacology/therapeutic use ; Dirofilaria immitis/drug effects/*microbiology ; Dirofilariasis/*drug therapy/*microbiology/transmission ; Doxycycline/pharmacology/*therapeutic use ; Drug Synergism ; Humans ; Ivermectin/therapeutic use ; Wolbachia/immunology/*physiology ; }, abstract = {Dirofilaria immitis, the cause of canine and feline heartworm disease, was the first filarial nematode described to harbour the bacterial endosymbiont Wolbachia. This ground-breaking discovery has led to intense research aimed at unravelling the nature of the endosymbiotic relationship; genomic studies have revealed how the bacteria may interact with the parasite and help explain why each is so dependent on the other. Analysis of the immune response to these bacteria may elucidate the mechanisms through which filarial parasites are able to survive for long periods of time in otherwise immune-competent hosts. Finally, studies aimed at the removal of the bacteria using specific antibiotic treatment in infected hosts is leading to the development of novel approaches for interrupting the transmission cycle and for the treatment and control of heartworm disease.}, } @article {pmid24812591, year = {2014}, author = {Hou, Q and He, J and Yu, J and Ye, Y and Zhou, D and Sun, Y and Zhang, D and Ma, L and Shen, B and Zhu, C}, title = {A case of horizontal gene transfer from Wolbachia to Aedes albopictus C6/36 cell line.}, journal = {Mobile genetic elements}, volume = {4}, number = {1}, pages = {e28914}, pmid = {24812591}, issn = {2159-2543}, support = {R01 AI075746/AI/NIAID NIH HHS/United States ; }, abstract = {Horizontal gene transfer plays an essential role in evolution and ecological adaptation, yet this phenomenon has remained controversial, particularly where it occurs between prokaryotes and eukaryotes. There are a handful of reported examples of horizontal gene transfer occurring between prokaryotes and eukaryotes in the literature, with most of these documented cases pertaining to invertebrates and endosymbionts. However, the vast majority of these horizontally transferred genes were either eventually excluded or rapidly became nonfunctional in the recipient genome. In this study, we report the discovery of a horizontal gene transfer from the endosymbiont Wolbachia in the C6/36 cell line derived from the mosquito Aedes albopictus. Moreover, we report that this horizontally transferred gene displayed high transcription level. This finding and the results of further experimentation strongly suggest this gene is functional and has been expressed and translated into a protein in the mosquito host cells.}, } @article {pmid24806619, year = {2014}, author = {Famah Sourassou, N and Hanna, R and Breeuwer, JA and Negloh, K and de Moraes, GJ and Sabelis, MW}, title = {The endosymbionts Wolbachia and Cardinium and their effects in three populations of the predatory mite Neoseiulus paspalivorus.}, journal = {Experimental & applied acarology}, volume = {64}, number = {2}, pages = {207-221}, pmid = {24806619}, issn = {1572-9702}, mesh = {Animals ; Anti-Bacterial Agents ; Benin ; Brazil ; DNA, Mitochondrial/genetics ; Female ; Genetic Variation ; Ghana ; Male ; Mites/genetics/*microbiology/*physiology ; Phylogeny ; Predatory Behavior ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Wolbachia/drug effects/*isolation & purification ; }, abstract = {Whereas endosymbiont-induced incompatibility is known to occur in various arthropod taxa, such as spider mites, insects and isopods, it has been rarely reported in plant-inhabiting predatory mites (Acari: Phytoseiidae). Recent cross-breeding studies with the phytoseiid mite Neoseiulus paspalivorus De Leon revealed a complete post-mating reproductive isolation between specimens collected from three geographic origins-Northeast Brazil (South America), Benin and Ghana (West Africa)-even though they are morphologically similar. We carried out a study to assess to what extent these populations exhibit genetic differences and whether endosymbionts are involved in the incompatibility. First, we used the mitochondrial cytochrome oxidase I (COI) gene to assess genetic diversity among the three populations. Second, we used a PCR-based method to check for the presence of Wolbachia and/or Cardinium in these populations, and we determined their phylogenetic relationships using specific primers for Wolbachia and Cardinium 16S rDNA genes. Third, we also conducted a test using an antibiotic (tetracycline) in an attempt to eliminate the symbionts and evaluate their effects on the reproductive compatibility of their host. Based on the DNA sequences of their COI genes, specimens of the three populations appear to be genetically similar. However, the 16S rDNA gene sequences of their associated endosymbionts differed among the three populations: the Benin and Brazil populations harbour different strains of Wolbachia symbionts, whereas the Ghana population harbours Cardinium symbionts. In response to antibiotic treatment females of each of the three populations became incompatible with untreated males of their own population, similar to that observed in crossings between females from one geographic population and males from another. Compatibility was restored in crosses involving uninfected Brazil females and uninfected Benin males, whereas the reciprocal crosses remained incompatible. Cardinium symbionts seem to be essential for oviposition in the Ghana population. It is concluded that their associated bacterial symbionts are the cause of the post-mating reproductive isolation previously observed among the three geographic populations. This insight is relevant to biological control of coconut mites for which N. paspalivorus is an effective predator, because introducing one geographic strain into the population of another (e.g. in field releases or mass cultures) may cause population growth depression.}, } @article {pmid24797695, year = {2014}, author = {Turley, AP and Smallegange, RC and Takken, W and Zalucki, MP and O'Neill, SL and McGraw, EA}, title = {Wolbachia infection does not alter attraction of the mosquito Aedes (Stegomyia) aegypti to human odours.}, journal = {Medical and veterinary entomology}, volume = {28}, number = {4}, pages = {457-460}, doi = {10.1111/mve.12063}, pmid = {24797695}, issn = {1365-2915}, mesh = {Aedes/*microbiology ; Aging ; Animals ; Feeding Behavior/*physiology ; Female ; Humans ; *Odorants ; Wolbachia/*isolation & purification ; }, abstract = {The insect endosymbiont Wolbachia pipientis (Rickettsiales: Rickettsiaceae) is undergoing field trials around the world to determine if it can reduce transmission of dengue virus from the mosquito Stegomyia aegypti to humans. Two different Wolbachia strains have been released to date. The primary effect of the wMel strain is pathogen protection whereby infection with the symbiont limits replication of dengue virus inside the mosquito. A second strain, wMelPop, induces pathogen protection, reduces the adult mosquito lifespan and decreases blood feeding success in mosquitoes after 15 days of age. Here we test whether Wolbachia infection affects mosquito attraction to host odours in adults aged 5 and 15 days. We found no evidence of reduced odour attraction of mosquitoes, even for those infected with the more virulent wMelPop. This bodes well for fitness and competitiveness in the field given that the mosquitoes must find hosts to reproduce for the biocontrol method to succeed.}, } @article {pmid24789726, year = {2014}, author = {Fallon, AM and Baldridge, GD and Carroll, EM and Kurtz, CM}, title = {Depletion of host cell riboflavin reduces Wolbachia levels in cultured mosquito cells.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {50}, number = {8}, pages = {707-713}, pmid = {24789726}, issn = {1543-706X}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/chemistry/drug effects/*microbiology ; Animals ; Cells, Cultured ; Culture Media ; Flavins/pharmacology ; Proteomics ; Riboflavin/analysis/*physiology ; *Wolbachia/genetics/growth & development ; }, abstract = {Wolbachia is an obligate intracellular alphaproteobacterium that occurs in arthropod and nematode hosts. Wolbachia presumably provides a fitness benefit to its hosts, but the basis for its retention and spread in host populations remains unclear. Wolbachia genomes retain biosynthetic pathways for some vitamins, and the possibility that these vitamins benefit host cells provides a potential means of selecting for Wolbachia-infected cell lines. To explore whether riboflavin produced by Wolbachia is available to its host cell, we established that growth of uninfected C7-10 mosquito cells decreases in riboflavin-depleted culture medium. A well-studied inhibitor of riboflavin uptake, lumiflavin, further inhibits growth of uninfected C7-10 cells with an LC50 of approximately 12 μg/ml. Growth of C/wStr1 mosquito cells, infected with Wolbachia from the planthopper, Laodelphax striatellus, was enhanced in medium containing low levels of lumiflavin, but Wolbachia levels decreased. Lumiflavin-enhanced growth thus resembled the improved growth that accompanies treatment with antibiotics that deplete Wolbachia, rather than a metabolic advantage provided by the Wolbachia infection. We used the polymerase chain reaction to validate the decrease in Wolbachia abundance and evaluated our results in the context of a proteomic analysis in which we detected nearly 800 wStr proteins. Our data indicate that Wolbachia converts riboflavin to FMN and FAD for its own metabolic needs, and does not provide a source of riboflavin for its host cell.}, } @article {pmid24789572, year = {2015}, author = {Hélène, D and Rémy, B and Nathalie, B and Anne-Laure, G and Traoré, RS and Jean-Michel, L and Bernard, R}, title = {Species and endosymbiont diversity of Bemisia tabaci (Homoptera: Aleyrodidae) on vegetable crops in Senegal.}, journal = {Insect science}, volume = {22}, number = {3}, pages = {386-398}, doi = {10.1111/1744-7917.12134}, pmid = {24789572}, issn = {1744-7917}, mesh = {Animals ; Bacteria/*genetics ; Biodiversity ; Crops, Agricultural ; DNA, Bacterial/genetics ; DNA, Mitochondrial/genetics ; Female ; Hemiptera/genetics/*microbiology ; Microsatellite Repeats ; Senegal ; Symbiosis ; Vegetables/parasitology ; }, abstract = {Bemisia tabaci-transmitted geminiviruses are one of the major threats on cassava and vegetable crops in Africa. However, to date, few studies are available on the diversity of B. tabaci and their associated endosymbionts in Africa. More than 28 species have been described in the complex of B. tabaci cryptic species; among them, 2 are invasive pests worldwide: MED and MEAM1. In order to assess the species diversity of B. tabaci in vegetable crops in Senegal, several samplings in different localities, hosts and seasons were collected and analyzed with nuclear (microsatellite) and mitochondrial (COI) markers. The bacterial endosymbiont community was also studied for each sample. Two species were detected: MED Q1 and MEAM1 B. Patterns of MED Q1 (dominance on most of the samples and sites, highest nuclear and mitochondrial diversity and broader secondary endosymbiont community: Hamiltonella, Cardinium, Wolbachia and Rickettsia), point toward a predominant resident begomovirus vector group for MED Q1 on market gardening crops. Furthermore, the lower prevalence of the second species MEAM1 B, its lower nuclear and mitochondrial diversity and a narrower secondary endosymbiont community (Hamiltonella/Rickettsia), indicate that this genetic group is exotic and results from a recent invasion in this area.}, } @article {pmid24787986, year = {2014}, author = {Zhong, Y and Li, ZX}, title = {Bidirectional cytoplasmic incompatibility induced by cross-order transfection of Wolbachia: implications for control of the host population.}, journal = {Microbial ecology}, volume = {68}, number = {3}, pages = {463-471}, pmid = {24787986}, issn = {1432-184X}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm/*microbiology ; DNA, Bacterial/genetics ; Female ; Fertility ; Hemiptera/*microbiology ; Male ; Pest Control, Biological ; Phenotype ; Phylogeny ; Sequence Analysis, DNA ; Symbiosis ; Wasps/*microbiology ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Wolbachia are widespread endosymbionts in arthropods and some nematodes. This genus of bacteria is known to manipulate host reproduction by inducing cytoplasmic incompatibility (CI). This important phenotype is implicated in the control of host populations since Wolbachia can suppress host populations through the induction of CI in a way similar to the sterile insect technique. Here, we identified a candidate CI-inducing Wolbachia strain from the parasitic wasp Scleroderma guani (wSguBJ) by sequencing and phylogenetic analysis. This Wolbachia strain was then isolated, purified, and artificially transfected into the new whitefly host Bemisia tabaci through nymphal microinjection. Infection frequency monitoring by molecular detection showed that 60-80 % of the offspring from transfected whitefly populations was infected with wSguBJ six generations after the transfer. Laboratory rearing experiments indicated that the artificial transfection caused no significant difference in the numbers of offspring between the transfected and naturally infected populations and had no significant detrimental effects on the development of transfected males, although the development of transfected females was delayed. Reciprocal crossings revealed that bidirectional CI was induced between the transfected and naturally infected whiteflies. These data indicated that the cross-order transfer of the heterologous Wolbachia strain by nymphal microinjection was successful. Mass release of the transfected males that could stably carry the heterologous Wolbachia without significant compromise of fecundity/development may provide an alternative approach to control of host populations.}, } @article {pmid24781809, year = {2014}, author = {Ma, WJ and Pannebakker, BA and Beukeboom, LW and Schwander, T and van de Zande, L}, title = {Genetics of decayed sexual traits in a parasitoid wasp with endosymbiont-induced asexuality.}, journal = {Heredity}, volume = {113}, number = {5}, pages = {424-431}, pmid = {24781809}, issn = {1365-2540}, mesh = {Animals ; Crosses, Genetic ; Female ; Genes, Recessive ; Male ; Reproduction, Asexual/*genetics ; Sex Ratio ; *Sexual Behavior, Animal ; Wasps/*genetics/*microbiology ; Wolbachia ; }, abstract = {Trait decay may occur when selective pressures shift, owing to changes in environment or life style, rendering formerly adaptive traits non-functional or even maladaptive. It remains largely unknown if such decay would stem from multiple mutations with small effects or rather involve few loci with major phenotypic effects. Here, we investigate the decay of female sexual traits, and the genetic causes thereof, in a transition from haplodiploid sexual reproduction to endosymbiont-induced asexual reproduction in the parasitoid wasp Asobara japonica. We take advantage of the fact that asexual females cured of their endosymbionts produce sons instead of daughters, and that these sons can be crossed with sexual females. By combining behavioral experiments with crosses designed to introgress alleles from the asexual into the sexual genome, we found that sexual attractiveness, mating, egg fertilization and plastic adjustment of offspring sex ratio (in response to variation in local mate competition) are decayed in asexual A. japonica females. Furthermore, introgression experiments revealed that the propensity for cured asexual females to produce only sons (because of decayed sexual attractiveness, mating behavior and/or egg fertilization) is likely caused by recessive genetic effects at a single locus. Recessive effects were also found to cause decay of plastic sex-ratio adjustment under variable levels of local mate competition. Our results suggest that few recessive mutations drive decay of female sexual traits, at least in asexual species deriving from haplodiploid sexual ancestors.}, } @article {pmid24779056, year = {2014}, author = {Dusi, E and Krenek, S and Schrallhammer, M and Sachse, R and Rauch, G and Kaltz, O and Berendonk, TU}, title = {Vertically transmitted symbiont reduces host fitness along temperature gradient.}, journal = {Journal of evolutionary biology}, volume = {27}, number = {4}, pages = {796-800}, doi = {10.1111/jeb.12336}, pmid = {24779056}, issn = {1420-9101}, mesh = {Gammaproteobacteria/*pathogenicity ; *Genetic Fitness ; Paramecium tetraurelia/*microbiology ; *Symbiosis ; *Temperature ; }, abstract = {Parasites with exclusive vertical transmission from host parent to offspring are an evolutionary puzzle. With parasite fitness entirely linked to host reproduction, any fitness cost for infected hosts risks their selective elimination. Environmental conditions likely influence parasite impact and thereby the success of purely vertical transmission strategies. We tested for temperature-dependent virulence of Caedibacter taeniospiralis, a vertically transmitted bacterial symbiont of the protozoan Paramecium tetraurelia. We compared growth of infected and cured host populations at five temperatures (16–32 °C). Infection reduced host density at all temperatures, with a peak of −30% at 28 °C. These patterns were largely consistent across five infected Paramecium strains. Similar to Wolbachia symbionts, C. taeniospiralis may compensate fitness costs by conferring to the host a ‘killer trait’, targeting uninfected competitors. Considerable loss of infection at 32 °C suggests that killer efficacy is not universal and that limited heat tolerance restricts the conditions for persistence of C. taeniospiralis.}, } @article {pmid24772084, year = {2014}, author = {Kosmidis, S and Missirlis, F and Botella, JA and Schneuwly, S and Rouault, TA and Skoulakis, EM}, title = {Behavioral decline and premature lethality upon pan-neuronal ferritin overexpression in Drosophila infected with a virulent form of Wolbachia.}, journal = {Frontiers in pharmacology}, volume = {5}, number = {}, pages = {66}, pmid = {24772084}, issn = {1663-9812}, abstract = {Iron is required for organismal growth. Therefore, limiting iron availability may be a key part of the host's innate immune response to various pathogens, for example, in Drosophila infected with Zygomycetes. One way the host can transiently reduce iron bioavailability is by ferritin overexpression. To study the effects of neuronal-specific ferritin overexpression on survival and neurodegeneration we generated flies simultaneously over-expressing transgenes for both ferritin subunits in all neurons. We used two independent recombinant chromosomes bearing UAS-Fer1HCH, UAS-Fer2LCH transgenes and obtained qualitatively different levels of late-onset behavioral and lifespan declines. We subsequently discovered that one parental strain had been infected with a virulent form of the bacterial endosymbiont Wolbachia, causing widespread neuronal apoptosis and premature death. This phenotype was exacerbated by ferritin overexpression and was curable by antibiotic treatment. Neuronal ferritin overexpression in uninfected flies did not cause evident neurodegeneration but resulted in a late-onset behavioral decline, as previously reported for ferritin overexpression in glia. The results suggest that ferritin overexpression in the central nervous system of flies is tolerated well in young individuals with adverse manifestations appearing only late in life or under unrelated pathophysiological conditions.}, } @article {pmid24763584, year = {2014}, author = {, }, title = {Genome sequence of the tsetse fly (Glossina morsitans): vector of African trypanosomiasis.}, journal = {Science (New York, N.Y.)}, volume = {344}, number = {6182}, pages = {380-386}, pmid = {24763584}, issn = {1095-9203}, support = {R01 AI081774/AI/NIAID NIH HHS/United States ; MR/K002279/1/MRC_/Medical Research Council/United Kingdom ; D43 TW007391/TW/FIC NIH HHS/United States ; R03 TW009444/TW/FIC NIH HHS/United States ; 098051/WT_/Wellcome Trust/United Kingdom ; 085775/Z/08/Z/WT_/Wellcome Trust/United Kingdom ; R01 AI051584/AI/NIAID NIH HHS/United States ; F32 CA091768/CA/NCI NIH HHS/United States ; U54 HG003079/HG/NHGRI NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; R03 TW008413/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; Blood ; Feeding Behavior ; Female ; Genes, Insect ; *Genome, Insect ; Insect Proteins/*genetics/physiology ; Insect Vectors/genetics/microbiology/parasitology/physiology ; Microbiota ; Molecular Sequence Annotation ; Molecular Sequence Data ; Reproduction/genetics ; Salivary Glands/parasitology/physiology ; Sensation/genetics ; Sequence Analysis, DNA ; Symbiosis ; Trypanosoma/physiology ; Trypanosomiasis, African/transmission ; Tsetse Flies/*genetics/microbiology/parasitology/physiology ; Wolbachia/genetics/physiology ; }, abstract = {Tsetse flies are the sole vectors of human African trypanosomiasis throughout sub-Saharan Africa. Both sexes of adult tsetse feed exclusively on blood and contribute to disease transmission. Notable differences between tsetse and other disease vectors include obligate microbial symbioses, viviparous reproduction, and lactation. Here, we describe the sequence and annotation of the 366-megabase Glossina morsitans morsitans genome. Analysis of the genome and the 12,308 predicted protein-encoding genes led to multiple discoveries, including chromosomal integrations of bacterial (Wolbachia) genome sequences, a family of lactation-specific proteins, reduced complement of host pathogen recognition proteins, and reduced olfaction/chemosensory associated genes. These genome data provide a foundation for research into trypanosomiasis prevention and yield important insights with broad implications for multiple aspects of tsetse biology.}, } @article {pmid24763283, year = {2014}, author = {Brelsfoard, C and Tsiamis, G and Falchetto, M and Gomulski, LM and Telleria, E and Alam, U and Doudoumis, V and Scolari, F and Benoit, JB and Swain, M and Takac, P and Malacrida, AR and Bourtzis, K and Aksoy, S}, title = {Presence of extensive Wolbachia symbiont insertions discovered in the genome of its host Glossina morsitans morsitans.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {4}, pages = {e2728}, pmid = {24763283}, issn = {1935-2735}, support = {R01 AI051584/AI/NIAID NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; AI068932/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Blotting, Southern ; *Genome, Bacterial ; *Genome, Insect ; In Situ Hybridization, Fluorescence ; Molecular Sequence Data ; *Mutagenesis, Insertional ; *Recombination, Genetic ; Sequence Analysis, DNA ; Tsetse Flies/*genetics ; Wolbachia/*genetics ; }, abstract = {Tsetse flies (Glossina spp.) are the cyclical vectors of Trypanosoma spp., which are unicellular parasites responsible for multiple diseases, including nagana in livestock and sleeping sickness in humans in Africa. Glossina species, including Glossina morsitans morsitans (Gmm), for which the Whole Genome Sequence (WGS) is now available, have established symbiotic associations with three endosymbionts: Wigglesworthia glossinidia, Sodalis glossinidius and Wolbachia pipientis (Wolbachia). The presence of Wolbachia in both natural and laboratory populations of Glossina species, including the presence of horizontal gene transfer (HGT) events in a laboratory colony of Gmm, has already been shown. We herein report on the draft genome sequence of the cytoplasmic Wolbachia endosymbiont (cytWol) associated with Gmm. By in silico and molecular and cytogenetic analysis, we discovered and validated the presence of multiple insertions of Wolbachia (chrWol) in the host Gmm genome. We identified at least two large insertions of chrWol, 527,507 and 484,123 bp in size, from Gmm WGS data. Southern hybridizations confirmed the presence of Wolbachia insertions in Gmm genome, and FISH revealed multiple insertions located on the two sex chromosomes (X and Y), as well as on the supernumerary B-chromosomes. We compare the chrWol insertions to the cytWol draft genome in an attempt to clarify the evolutionary history of the HGT events. We discuss our findings in light of the evolution of Wolbachia infections in the tsetse fly and their potential impacts on the control of tsetse populations and trypanosomiasis.}, } @article {pmid24763092, year = {2014}, author = {Chiel, E and Kelly, SE and Harris, AM and Gebiola, M and Li, X and Zchori-Fein, E and Hunter, MS}, title = {Characteristics, phenotype, and transmission of Wolbachia in the sweet potato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), and its parasitoid Eretmocerus sp. nr. emiratus (Hymenoptera: Aphelinidae).}, journal = {Environmental entomology}, volume = {43}, number = {2}, pages = {353-362}, doi = {10.1603/EN13286}, pmid = {24763092}, issn = {1938-2936}, mesh = {Animals ; Computer Simulation ; Crosses, Genetic ; *Disease Transmission, Infectious ; Female ; Hemiptera/genetics/*microbiology/*parasitology/physiology ; Ipomoea batatas/parasitology ; Likelihood Functions ; Male ; Models, Biological ; *Phenotype ; Polymerase Chain Reaction ; Reproduction/genetics ; Species Specificity ; Symbiosis ; Wasps/genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a common intracellular bacterial endosymbiont of insects, causing a variety of effects including reproductive manipulations such as cytoplasmic incompatibility (CI). In this study, we characterized Wolbachia in the whitefly Bemisia tabaci and in the whitefly parasitoid Eretmocerus sp. nr. emiratus. We also tested for horizontal transmission of Wolbachia between and within trophic levels, and we determined the phenotype of Wolbachia in E. sp. nr. emiratus. Using multilocus sequence typing and phylogenetic analyses, we found that B. tabaci and E. sp. nr. emiratus each harbor a different and unique strain of Wolbachia. Both strains belong to the phylogenetic supergroup B. No evidence for horizontal transmission of Wolbachia between and within trophic levels was found in our study system. Finally, crossing results were consistent with a CI phenotype; when Wolbachia-infected E. sp. nr. emiratus males mate with uninfected females, wasp progeny survival dropped significantly, and the number of females was halved. This is the first description of CI caused by Wolbachia in the economically important genus Eretmocerus. Our study underscores the expectation that horizontal transmission events occur rarely in the dynamics of secondary symbionts such as Wolbachia, and highlights the importance of understanding the effects of symbionts on the biology of natural enemies.}, } @article {pmid24759973, year = {2014}, author = {Telschow, A and Hilgenboecker, K and Hammerstein, P and Werren, JH}, title = {Dobzhansky-muller and wolbachia-induced incompatibilities in a diploid genetic system.}, journal = {PloS one}, volume = {9}, number = {4}, pages = {e95488}, pmid = {24759973}, issn = {1932-6203}, mesh = {Animals ; *Diploidy ; *Genetic Speciation ; Genetic Variation ; Hybridization, Genetic ; Insecta/microbiology ; Wolbachia/*physiology ; }, abstract = {Genetic incompatibilities are supposed to play an important role in speciation. A general (theoretical) problem is to explain the persistence of genetic diversity after secondary contact. Previous theoretical work has pointed out that Dobzhansky-Muller incompatibilities (DMI) are not stable in the face of migration unless local selection acts on the alleles involved in incompatibility. With local selection, genetic variability exists up to a critical migration rate but is lost when migration exceeds this threshold value. Here, we investigate the effect of intracellular bacteria Wolbachia on the stability of hybrid zones formed after the Dobzhansky Muller model. Wolbachia are known to cause a cytoplasmic incompatibility (CI) within and between species. Incorporating intracellular bacteria Wolbachia can lead to a significant increase of critical migration rates and maintenance of divergence, primarily because Wolbachia-induced incompatibility acts to reduce frequencies of F1 hybrids. Wolbachia infect up to two-thirds of all insect species and it is therefore likely that CI co-occurs with DMI in nature. The results indicate that both isolating mechanisms strengthen each other and under some circumstances act synergistically. Thus they can drive speciation processes more forcefully than either when acting alone.}, } @article {pmid24759922, year = {2014}, author = {Mayoral, JG and Etebari, K and Hussain, M and Khromykh, AA and Asgari, S}, title = {Wolbachia infection modifies the profile, shuttling and structure of microRNAs in a mosquito cell line.}, journal = {PloS one}, volume = {9}, number = {4}, pages = {e96107}, pmid = {24759922}, issn = {1932-6203}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Argonaute Proteins/genetics ; Cell Line ; Cell Nucleus/*genetics ; Cytoplasm/*genetics ; Gene Expression Regulation ; High-Throughput Nucleotide Sequencing ; Host-Pathogen Interactions ; Insect Proteins/genetics ; MicroRNAs/chemistry/*genetics ; Molecular Sequence Data ; RNA Transport ; Wolbachia/*physiology ; }, abstract = {MicroRNAs (miRNAs) are small non-coding RNAs that play important roles in many biological processes such as development, cell signaling and immune response. Small RNA deep sequencing technology provided an opportunity for a thorough survey of the miRNA profile of a mosquito cell line from Aedes aegypti. We characterized the miRNA composition of the nucleus and the cytoplasm of uninfected cells and compared it with the one of cells infected with the endosymbiotic bacterium Wolbachia strain wMelPop-CLA. We found an overall increase of small RNAs between 18 and 28 nucleotides in both cellular compartments in Wolbachia-infected cells and identified specific miRNAs induced and/or suppressed by the Wolbachia infection. We discuss the mechanisms that the cell may use to shuttle miRNAs between the cytoplasm and the nucleus. In addition, we identified piRNAs that changed their abundance in response to Wolbachia infection. The miRNAs and piRNAs identified in this study provide promising leads for investigations into the host-endosymbiont interactions and for better understanding of how Wolbachia manipulates the host miRNA machinery in order to facilitate its persistent replication in infected cells.}, } @article {pmid24750488, year = {2014}, author = {Dittmer, J and Beltran-Bech, S and Lesobre, J and Raimond, M and Johnson, M and Bouchon, D}, title = {Host tissues as microhabitats for Wolbachia and quantitative insights into the bacterial community in terrestrial isopods.}, journal = {Molecular ecology}, volume = {23}, number = {10}, pages = {2619-2635}, doi = {10.1111/mec.12760}, pmid = {24750488}, issn = {1365-294X}, mesh = {Animals ; Bacterial Load ; *Biological Evolution ; Female ; Genetics, Population ; Isopoda/*microbiology ; Male ; Microbiota ; Microsatellite Repeats ; Sequence Analysis, DNA ; Symbiosis/*genetics ; Wolbachia/genetics/*physiology ; }, abstract = {Animal-bacterial symbioses are highly dynamic in terms of multipartite interactions, both between the host and its symbionts as well as between the different bacteria constituting the symbiotic community. These interactions will be reflected by the titres of the individual bacterial taxa, for example via host regulation of bacterial loads or competition for resources between symbionts. Moreover, different host tissues represent heterogeneous microhabitats for bacteria, meaning that host-associated bacteria might establish tissue-specific bacterial communities. Wolbachia are widespread endosymbiotic bacteria, infecting a large number of arthropods and filarial nematodes. However, relatively little is known regarding direct interactions between Wolbachia and other bacteria. This study represents the first quantitative investigation of tissue-specific Wolbachia-microbiota interactions in the terrestrial isopod Armadillidium vulgare. To this end, we obtained a more complete picture of the Wolbachia distribution patterns across all major host tissues, integrating all three feminizing Wolbachia strains (wVulM, wVulC, wVulP) identified to date in this host. Interestingly, the different Wolbachia strains exhibited strain-specific tissue distribution patterns, with wVulM reaching lower titres in most tissues. These patterns were consistent across different host genetic backgrounds and might reflect different co-evolutionary histories between the Wolbachia strains and A. vulgare. Moreover, Wolbachia-infected females carried higher total bacterial loads in several, but not all, tissues, irrespective of the Wolbachia strain. Taken together, this quantitative approach indicates that Wolbachia is part of a potentially more diverse bacterial community, as exemplified by the presence of highly abundant bacterial taxa in the midgut caeca of several A. vulgare populations.}, } @article {pmid24747216, year = {2014}, author = {Kawasaki, Y and Ito, H and Kajimura, H}, title = {Equilibrium frequency of endosymbionts in multiple infections based on the balance between vertical transmission and cytoplasmic incompatibility.}, journal = {PloS one}, volume = {9}, number = {4}, pages = {e94900}, pmid = {24747216}, issn = {1932-6203}, mesh = {*Bacteria ; *Bacterial Infections ; *Bacterial Physiological Phenomena ; Cytoplasm/*microbiology ; *Models, Biological ; Reproduction ; *Symbiosis ; }, abstract = {Cytoplasmic incompatibility (CI)-inducing endosymbiotic bacteria, such as Wolbachia and Cardinium, have been well studied through field data and validations on the basis of numerical simulations. However, the analytically derived equilibrium frequency of multiple infections has not yet been determined, although the equilibrium for cases of single infection has been reported. In this study, we considered the difference equation for endosymbionts using three parameters: the probability of the failure of vertical transmission ([Formula: see text]), CI strength ([Formula: see text]), and the level of host inbreeding ([Formula: see text]). To analyze this model, we particularly focused on [Formula: see text], i.e., the frequency of host individuals completely infected with all [Formula: see text]-bacterial strains in the population. [Formula: see text], [Formula: see text] at the equilibrium state, was analytically calculated in the cases where [Formula: see text] and [Formula: see text] is any arbitrary value. We found that [Formula: see text] can be described using two parameters: [Formula: see text] and [Formula: see text], which is identical to [Formula: see text]. [Formula: see text] has a larger value in a system with a smaller [Formula: see text]. In addition, [Formula: see text] determines the maximum number of strains that infect a single host. Our results revealed the following: i) three parameters can be reduced to a single parameter, i.e., [Formula: see text] and ii) the threshold of the maximum number of infections is defined by [Formula: see text], which prevents additional invasions by endosymbionts.}, } @article {pmid24732463, year = {2014}, author = {Ross, PA and Endersby, NM and Yeap, HL and Hoffmann, AA}, title = {Larval competition extends developmental time and decreases adult size of wMelPop Wolbachia-infected Aedes aegypti.}, journal = {The American journal of tropical medicine and hygiene}, volume = {91}, number = {1}, pages = {198-205}, pmid = {24732463}, issn = {1476-1645}, mesh = {Aedes/growth & development/*microbiology ; Animals ; Biological Control Agents ; Female ; Insect Vectors/growth & development/*microbiology ; Larva/growth & development/*microbiology ; Male ; Pest Control, Biological/*methods ; Wolbachia/*pathogenicity/physiology ; }, abstract = {The intracellular endosymbiont Wolbachia has been artificially transinfected into the dengue vector Aedes aegypti, where it is being investigated as a potential dengue biological control agent. Invasion of Wolbachia in natural populations depends upon the fitness of Wolbachia-infected Ae. aegypti relative to uninfected competitors. Although Wolbachia infections impose fitness costs on the adult host, effects at the immature stages are less clear, particularly in competitive situations. We look for effects of two Wolbachia infections, wMel and wMelPop, on intra-strain and inter-strain larval competition in Ae. aegypti. Development of Wolbachia-infected larvae is delayed in mixed cohorts with uninfected larvae under crowded-rearing conditions. Slow developing wMelPop-infected larvae have reduced adult size compared with uninfected larvae, and larvae with the wMel infection are somewhat larger and have greater viability relative to uninfected larvae when in mixed cohorts. Implications for successful invasion by these Wolbachia infections under field conditions are considered.}, } @article {pmid24722673, year = {2014}, author = {Genty, LM and Bouchon, D and Raimond, M and Bertaux, J}, title = {Wolbachia infect ovaries in the course of their maturation: last minute passengers and priority travellers?.}, journal = {PloS one}, volume = {9}, number = {4}, pages = {e94577}, pmid = {24722673}, issn = {1932-6203}, mesh = {Animals ; Bacterial Load ; Cell Size ; Female ; Host-Pathogen Interactions ; In Situ Hybridization, Fluorescence ; Isopoda/growth & development/*microbiology ; Oocytes/growth & development/*microbiology ; Ovary/growth & development/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are widespread endosymbiotic bacteria of arthropods and nematodes. Studies on such models suggest that Wolbachia's remarkable aptitude to infect offspring may rely on a re-infection of ovaries from somatic tissues instead of direct cellular segregation between oogonia and oocytes. In the terrestrial isopod Armadillidium vulgare, Wolbachia are vertically transmitted to the host offspring, even though ovary cells are cyclically renewed. Using Fluorescence in situ hybridization (FISH), we showed that the proportion of infected oocytes increased in the course of ovary and oocyte maturation, starting with 31.5% of infected oocytes only. At the end of ovary maturation, this proportion reached 87.6% for the most mature oocytes, which is close to the known transmission rate to offspring. This enrichment can be explained by a secondary acquisition of the bacteria by oocytes (Wolbachia can be seen as last minute passengers) and/or by a preferential selection of oocytes infected with Wolbachia (as priority travellers).}, } @article {pmid24721205, year = {2014}, author = {Liu, C and Wang, JL and Zheng, Y and Xiong, EJ and Li, JJ and Yuan, LL and Yu, XQ and Wang, YF}, title = {Wolbachia-induced paternal defect in Drosophila is likely by interaction with the juvenile hormone pathway.}, journal = {Insect biochemistry and molecular biology}, volume = {49}, number = {}, pages = {49-58}, doi = {10.1016/j.ibmb.2014.03.014}, pmid = {24721205}, issn = {1879-0240}, mesh = {Animals ; Drosophila Proteins/genetics/*metabolism ; Drosophila melanogaster/embryology/*genetics/metabolism/*microbiology ; Female ; Juvenile Hormones/*metabolism ; Male ; Ovum/metabolism/microbiology ; Signal Transduction ; Species Specificity ; Testis/metabolism/microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbionts that infect many insect species. They can manipulate the host's reproduction to increase their own maternal transmission. Cytoplasmic incompatibility (CI) is one such manipulation, which is expressed as embryonic lethality when Wolbachia-infected males mate with uninfected females. However, matings between males and females carrying the same Wolbachia strain result in viable progeny. The molecular mechanisms of CI are currently not clear. We have previously reported that the gene Juvenile hormone-inducible protein 26 (JhI-26) exhibited the highest upregulation in the 3rd instar larval testes of Drosophila melanogaster when infected by Wolbachia. This is reminiscent of an interaction between Wolbachia and juvenile hormone (JH) pathway in flies. Considering that Jhamt gene encodes JH acid methyltransferase, a key regulatory enzyme of JH biosynthesis, and that methoprene-tolerant (Met) has been regarded as the best JH receptor candidate, we first compared the expression of Jhamt and Met between Wolbachia-infected and uninfected fly testes to investigate whether Wolbachia infection influence the JH signaling pathway. We found that the expressions of Jhamt and Met were significantly increased in the presence of Wolbachia, suggesting an interaction of Wolbachia with the JH signaling pathway. Then, we found that overexpression of JhI-26 in Wolbachia-free transgenic male flies caused paternal-effect lethality that mimics the defects associated with CI. JhI-26 overexpressing males resulted in significantly decrease in hatch rate. Surprisingly, Wolbachia-infected females could rescue the egg hatch. In addition, we showed that overexpression of JhI-26 caused upregulation of the male accessory gland protein (Acp) gene CG10433, but not vice versa. This result suggests that JhI-26 may function at the upstream of CG10433. Likewise, overexpression of CG10433 also resulted in paternal-effect lethality. Both JhI-26 and CG10433 overexpressing males resulted in nuclear division defects in the early embryos. Finally, we found that Wolbachia-infected males decreased the propensity of the mated females to remating, a phenotype also caused by both JhI-26 and CG10433 overexpressing males. Taken together, our results provide a working hypothesis whereby Wolbachia induce paternal defects in Drosophila probably by interaction with the JH pathway via JH response genes JhI-26 and CG10433.}, } @article {pmid24714809, year = {2014}, author = {Huang, W and Massouras, A and Inoue, Y and Peiffer, J and Ràmia, M and Tarone, AM and Turlapati, L and Zichner, T and Zhu, D and Lyman, RF and Magwire, MM and Blankenburg, K and Carbone, MA and Chang, K and Ellis, LL and Fernandez, S and Han, Y and Highnam, G and Hjelmen, CE and Jack, JR and Javaid, M and Jayaseelan, J and Kalra, D and Lee, S and Lewis, L and Munidasa, M and Ongeri, F and Patel, S and Perales, L and Perez, A and Pu, L and Rollmann, SM and Ruth, R and Saada, N and Warner, C and Williams, A and Wu, YQ and Yamamoto, A and Zhang, Y and Zhu, Y and Anholt, RR and Korbel, JO and Mittelman, D and Muzny, DM and Gibbs, RA and Barbadilla, A and Johnston, JS and Stone, EA and Richards, S and Deplancke, B and Mackay, TF}, title = {Natural variation in genome architecture among 205 Drosophila melanogaster Genetic Reference Panel lines.}, journal = {Genome research}, volume = {24}, number = {7}, pages = {1193-1208}, pmid = {24714809}, issn = {1549-5469}, support = {R01 GM059469/GM/NIGMS NIH HHS/United States ; U54 HG003273/HG/NHGRI NIH HHS/United States ; R01 AA016560/AA/NIAAA NIH HHS/United States ; R01 GM045146/GM/NIGMS NIH HHS/United States ; R01 GM076083/GM/NIGMS NIH HHS/United States ; R01GM076083/GM/NIGMS NIH HHS/United States ; R01 GM45146/GM/NIGMS NIH HHS/United States ; R01 GM 59469/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Chromatin/genetics/metabolism ; Drosophila melanogaster/*genetics/microbiology ; Female ; Genetic Linkage ; *Genetic Variation ; Genome Size ; *Genome, Insect ; Genome-Wide Association Study ; Genotype ; Genotyping Techniques ; High-Throughput Nucleotide Sequencing ; INDEL Mutation ; Linkage Disequilibrium ; Male ; Molecular Sequence Annotation ; *Phenotype ; Polymorphism, Single Nucleotide ; Quantitative Trait, Heritable ; Reproducibility of Results ; }, abstract = {The Drosophila melanogaster Genetic Reference Panel (DGRP) is a community resource of 205 sequenced inbred lines, derived to improve our understanding of the effects of naturally occurring genetic variation on molecular and organismal phenotypes. We used an integrated genotyping strategy to identify 4,853,802 single nucleotide polymorphisms (SNPs) and 1,296,080 non-SNP variants. Our molecular population genomic analyses show higher deletion than insertion mutation rates and stronger purifying selection on deletions. Weaker selection on insertions than deletions is consistent with our observed distribution of genome size determined by flow cytometry, which is skewed toward larger genomes. Insertion/deletion and single nucleotide polymorphisms are positively correlated with each other and with local recombination, suggesting that their nonrandom distributions are due to hitchhiking and background selection. Our cytogenetic analysis identified 16 polymorphic inversions in the DGRP. Common inverted and standard karyotypes are genetically divergent and account for most of the variation in relatedness among the DGRP lines. Intriguingly, variation in genome size and many quantitative traits are significantly associated with inversions. Approximately 50% of the DGRP lines are infected with Wolbachia, and four lines have germline insertions of Wolbachia sequences, but effects of Wolbachia infection on quantitative traits are rarely significant. The DGRP complements ongoing efforts to functionally annotate the Drosophila genome. Indeed, 15% of all D. melanogaster genes segregate for potentially damaged proteins in the DGRP, and genome-wide analyses of quantitative traits identify novel candidate genes. The DGRP lines, sequence data, genotypes, quality scores, phenotypes, and analysis and visualization tools are publicly available.}, } @article {pmid24714653, year = {2014}, author = {Das, B and Satapathy, T and Kar, SK and Hazra, RK}, title = {Genetic structure and Wolbachia genotyping in naturally occurring populations of Aedes albopictus across contiguous landscapes of Orissa, India.}, journal = {PloS one}, volume = {9}, number = {4}, pages = {e94094}, pmid = {24714653}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Animals ; Chikungunya Fever/transmission ; Dengue/transmission ; *Genetic Variation ; *Genotype ; India ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Aedes albopictus has recently been implicated as a major vector in the emergence of dengue and chikungunya in several parts of India, like Orissa, which is gradually gaining endemicity for arboviral diseases. Ae. albopictus is further known to be naturally infected with Wolbachia (maternally inherited bacterium), which causes cytoplasmic incompatibility (CI) in mosquitoes leading to sperm-egg incompatibility inducing the death of embryo. Knowledge of genetic diversity of Ae. albopictus, along with revealing the type of Wolbachia infection in Ae. albopictus is important to explore the genetic and biological characteristics of Ae. albopictus, prior to exploring the uses of CI-based vector control strategies. In this study, we assessed the population genetic structure and the pattern of Wolbachia infection in Ae. albopictus mosquitoes of Orissa.

METHODS AND RESULTS: Ae. albopictus mosquitoes were collected from 15 districts representing the four physiographical regions of Orissa from 2010-2012, analyzed for genetic variability at seven microsatellite loci and genotyped for Wolbachia strain detection using wsp gene primers. Most microsatellite markers were successfully amplified and were polymorphic, showing moderate genetic structure among all geographic populations (FST = 0.088). Genetic diversity was high (FST = 0.168) in Coastal Plains populations when compared with other populations, which was also evident from cluster analyses that showed most Coastal Plains populations consisted of a separate genetic cluster. Genotyping analyses revealed that Wolbachia-infected Ae. albopictus field populations of Orissa were mostly superinfected with wAlbA and wAlbB strains. Wolbachia superinfection was more pronounced in the Coastal Plain populations.

CONCLUSION: High genetic structure and Wolbachia superinfection, observed in the Coastal Plain populations of Orissa suggested it to be genetically and biologically more unique than other populations, and hence could influence their vectorial attributes. Such high genetic diversity observed among Coastal Plains populations could be attributed to multiple introductions of Ae. albopictus in this region.}, } @article {pmid24697073, year = {2013}, author = {Chai, HN and Wu, HY and Du, YZ}, title = {[Detection and analysis of symbiotic bacteria, Arsenophonus and Wolbachia, in striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae)].}, journal = {Ying yong sheng tai xue bao = The journal of applied ecology}, volume = {24}, number = {12}, pages = {3517-3522}, pmid = {24697073}, issn = {1001-9332}, mesh = {Animals ; China ; DNA, Bacterial/genetics ; Enterobacteriaceae/*classification/isolation & purification ; Genes, Bacterial ; Lepidoptera/*microbiology ; *Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 23S/genetics ; *Symbiosis ; Wolbachia/*classification/isolation & purification ; }, abstract = {In this study, 23S rDNA from Arsenophonus and wsp gene from Wolbachia were amplified by PCR method using specific primers to study their symbiosis in Chilo suppressalis from 20 locations of China. The results indicated that Arsenophonus and Wolbachia infection rates differed widely in C. suppressalis populations. The infection rates of Arsenophonus ranged from 5.0% to 50.0% in five geographical populations from Harbin, Huishui, Jilin, Nanyang, and Yangzhou. Wolbachia ranged from 25.0% to 40.0% in three geographical populations from Hanzhong, Nanning, and Yangzhou. Their symbiosis was not observed in C. suppressalis of other locations. The sequences of 23S rDNA gene which named csArs were exactly the same in the five populations. However, the sequences of wsp from the three strains of Wolbachia showed wChisup1 belonged to supergroup A, wChisup5 and wChisup6 belonged to supergroup B. The results showed the strains of Arsenophonus from C. suppressalis of the five different locations were identical, whereas the strains of Wolbachia were phylogenetically diverse. The phylogenetic analysis indicated that 23S rDNA and wsp sequences found in C. suppressalis were exactly the same with or very closely related to relevant sequences in other species.}, } @article {pmid24693489, year = {2014}, author = {Franz, AW and Clem, RJ and Passarelli, AL}, title = {Novel Genetic and Molecular Tools for the Investigation and Control of Dengue Virus Transmission by Mosquitoes.}, journal = {Current tropical medicine reports}, volume = {1}, number = {1}, pages = {21-31}, pmid = {24693489}, issn = {2196-3045}, support = {R01 AI091972/AI/NIAID NIH HHS/United States ; }, abstract = {Aedes aegypti is the principal vector of dengue virus (DENV) throughout the tropical world. This anthropophilic mosquito species needs to be persistently infected with DENV before it can transmit the virus through its saliva to a new vertebrate host. In the mosquito, DENV is confronted with several innate immune pathways, among which RNA interference is considered the most important. The Ae. aegypti genome project opened the doors for advanced molecular studies on pathogen-vector interactions including genetic manipulation of the vector for basic research and vector control purposes. Thus, Ae. aegypti has become the primary model for studying vector competence for arboviruses at the molecular level. Here, we present recent findings regarding DENV-mosquito interactions, emphasizing how innate immune responses modulate DENV infections in Ae. aegypti. We also describe the latest advancements in genetic manipulation of Ae. aegypti and discuss how this technology can be used to investigate vector transmission of DENV at the molecular level and to control transmission of the virus in the field.}, } @article {pmid24686935, year = {2014}, author = {Normark, BB and Ross, L}, title = {Genetic conflict, kin and the origins of novel genetic systems.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {369}, number = {1642}, pages = {20130364}, pmid = {24686935}, issn = {1471-2970}, mesh = {Adaptation, Biological/*genetics ; Animals ; *Biological Evolution ; *Environment ; Female ; *Genetic Fitness ; *Genetics, Population ; Germ Cells/physiology ; Inheritance Patterns/*genetics ; Male ; *Models, Genetic ; Ploidies ; Sex Factors ; Symbiosis ; }, abstract = {Genetic conflict may have played an important role in the evolution of novel genetic systems. The ancestral system of eumendelian genetics is highly symmetrical. Those derived from it (e.g. thelytokous parthenogenesis, haplodiploidy and parent-specific allele expression) are more asymmetrical in the genetic role played by maternal versus paternal alleles. These asymmetries may have arisen from maternal-paternal genetic conflict, or cytonuclear conflict, or from an interaction between them. Asymmetric genetic systems are much more common in terrestrial and freshwater taxa than in marine taxa. We suggest three reasons for this, based on the relative inhospitability of terrestrial environments to three types of organism: (i) pathogens-departure from the marine realm meant escape from many pathogens and parasites, reducing the need for sexual reproduction; (ii) symbionts-symbionts are no more important in the terrestrial realm than the marine realm but are more likely to be obligately intracellular and vertically transmitted, making them more likely to disrupt their host's genetic systems; (iii) Gametes and embryos-because neither gametes nor embryos can be shed into air as easily as into seawater, the mother's body is a more important environment for both types of organisms in the terrestrial realm than in the marine realm. This environment of asymmetric kinship (with neighbours more closely related by maternal alleles than by paternal alleles) may have helped to drive asymmetries in expression and transmission.}, } @article {pmid24685011, year = {2014}, author = {Foster, JM and Landmann, F and Ford, L and Johnston, KL and Elsasser, SC and Schulte-Hostedde, AI and Taylor, MJ and Slatko, BE}, title = {Absence of Wolbachia endobacteria in the human parasitic nematode Dracunculus medinensis and two related Dracunculus species infecting wildlife.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {140}, pmid = {24685011}, issn = {1756-3305}, mesh = {Animals ; Animals, Wild ; DNA, Bacterial/genetics/isolation & purification ; Dracunculiasis/epidemiology/parasitology/*veterinary ; Dracunculus Nematode/*microbiology ; Female ; Ghana/epidemiology ; Humans ; Mink/parasitology ; Ontario/epidemiology ; Otters/parasitology ; Species Specificity ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia endosymbionts are a proven target for control of human disease caused by filarial nematodes. However, little is known about the occurrence of Wolbachia in taxa closely related to the superfamily Filarioidea. Our study addressed the status of Wolbachia presence in members of the superfamily Dracunculoidea by screening the human parasite Dracunculus medinensis and related species from wildlife for Wolbachia.

FINDINGS: D. medinensis, D. lutrae and D. insignis specimens were all negative for Wolbachia colonization by PCR screening for the Wolbachia ftsZ, 16S rRNA and Wolbachia surface protein (wsp) sequences. The quality and purity of the DNA preparations was confirmed by amplification of nematode 18S rRNA and cytochrome c oxidase subunit I sequences. Furthermore, Wolbachia endobacteria were not detected by whole mount fluorescence staining, or by immunohistochemistry using a Wolbachia-specific antiserum. In contrast, positive control Brugia malayi worms were shown to harbour Wolbachia by PCR, fluorescence staining and immunohistochemistry.

CONCLUSIONS: Three examined species of Dracunculus showed no evidence of Wolbachia endobacteria. This supports that members of the superfamily Dracunculoidea are free of Wolbachia. Within the order Spirurida, these endosymbionts appear restricted to the Filarioidea.}, } @article {pmid24673824, year = {2014}, author = {Boivin, T and Henri, H and Vavre, F and Gidoin, C and Veber, P and Candau, JN and Magnoux, E and Roques, A and Auger-Rozenberg, MA}, title = {Epidemiology of asexuality induced by the endosymbiotic Wolbachia across phytophagous wasp species: host plant specialization matters.}, journal = {Molecular ecology}, volume = {23}, number = {9}, pages = {2362-2375}, doi = {10.1111/mec.12737}, pmid = {24673824}, issn = {1365-294X}, mesh = {Animals ; Bacterial Typing Techniques ; Bayes Theorem ; Female ; Genetics, Population ; Likelihood Functions ; Molecular Sequence Data ; Multilocus Sequence Typing ; *Parthenogenesis ; Phylogeny ; *Symbiosis ; Wasps/*microbiology ; Wolbachia/classification/*genetics ; }, abstract = {Among eukaryotes, sexual reproduction is by far the most predominant mode of reproduction. However, some systems maintaining sexuality appear particularly labile and raise intriguing questions on the evolutionary routes to asexuality. Thelytokous parthenogenesis is a form of spontaneous loss of sexuality leading to strong distortion of sex ratio towards females and resulting from mutation, hybridization or infection by bacterial endosymbionts. We investigated whether ecological specialization is a likely mechanism of spread of thelytoky within insect communities. Focusing on the highly specialized genus Megastigmus (Hymenoptera: Torymidae), we first performed a large literature survey to examine the distribution of thelytoky in these wasps across their respective obligate host plant families. Second, we tested for thelytoky caused by endosymbionts by screening in 15 arrhenotokous and 10 thelytokous species for Wolbachia, Cardinium, Arsenophonus and Rickettsia endosymbionts and by performing antibiotic treatments. Finally, we performed phylogenetic reconstructions using multilocus sequence typing (MLST) to examine the evolution of endosymbiont-mediated thelytoky in Megastigmus and its possible connections to host plant specialization. We demonstrate that thelytoky evolved from ancestral arrhenotoky through the horizontal transmission and the fixation of the parthenogenesis-inducing Wolbachia. We find that ecological specialization in Wolbachia's hosts was probably a critical driving force for Wolbachia infection and spread of thelytoky, but also a constraint. Our work further reinforces the hypothesis that community structure of insects is a major driver of the epidemiology of endosymbionts and that competitive interactions among closely related species may facilitate their horizontal transmission.}, } @article {pmid24641208, year = {2014}, author = {Shokralla, S and Gibson, JF and Nikbakht, H and Janzen, DH and Hallwachs, W and Hajibabaei, M}, title = {Next-generation DNA barcoding: using next-generation sequencing to enhance and accelerate DNA barcode capture from single specimens.}, journal = {Molecular ecology resources}, volume = {14}, number = {5}, pages = {892-901}, pmid = {24641208}, issn = {1755-0998}, mesh = {Animals ; Cost-Benefit Analysis ; Costa Rica ; DNA Barcoding, Taxonomic/*methods ; High-Throughput Nucleotide Sequencing/*methods ; Lepidoptera/classification/genetics ; Time Factors ; }, abstract = {DNA barcoding is an efficient method to identify specimens and to detect undescribed/cryptic species. Sanger sequencing of individual specimens is the standard approach in generating large-scale DNA barcode libraries and identifying unknowns. However, the Sanger sequencing technology is, in some respects, inferior to next-generation sequencers, which are capable of producing millions of sequence reads simultaneously. Additionally, direct Sanger sequencing of DNA barcode amplicons, as practiced in most DNA barcoding procedures, is hampered by the need for relatively high-target amplicon yield, coamplification of nuclear mitochondrial pseudogenes, confusion with sequences from intracellular endosymbiotic bacteria (e.g. Wolbachia) and instances of intraindividual variability (i.e. heteroplasmy). Any of these situations can lead to failed Sanger sequencing attempts or ambiguity of the generated DNA barcodes. Here, we demonstrate the potential application of next-generation sequencing platforms for parallel acquisition of DNA barcode sequences from hundreds of specimens simultaneously. To facilitate retrieval of sequences obtained from individual specimens, we tag individual specimens during PCR amplification using unique 10-mer oligonucleotides attached to DNA barcoding PCR primers. We employ 454 pyrosequencing to recover full-length DNA barcodes of 190 specimens using 12.5% capacity of a 454 sequencing run (i.e. two lanes of a 16 lane run). We obtained an average of 143 sequence reads for each individual specimen. The sequences produced are full-length DNA barcodes for all but one of the included specimens. In a subset of samples, we also detected Wolbachia, nontarget species, and heteroplasmic sequences. Next-generation sequencing is of great value because of its protocol simplicity, greatly reduced cost per barcode read, faster throughout and added information content.}, } @article {pmid24627993, year = {2014}, author = {van der Kooi, CJ and Schwander, T}, title = {Evolution of asexuality via different mechanisms in grass thrips (thysanoptera: Aptinothrips).}, journal = {Evolution; international journal of organic evolution}, volume = {68}, number = {7}, pages = {1883-1893}, doi = {10.1111/evo.12402}, pmid = {24627993}, issn = {1558-5646}, mesh = {Animals ; *Evolution, Molecular ; Female ; Reproduction, Asexual/*genetics ; Symbiosis ; Thysanoptera/*genetics/microbiology/physiology ; Wolbachia/pathogenicity ; }, abstract = {Asexual lineages can derive from sexual ancestors via different mechanisms and at variable rates, which affects the diversity of the asexual population and thereby its ecological success. We investigated the variation and evolution of reproductive systems in Aptinothrips, a genus of grass thrips comprising four species. Extensive population surveys and breeding experiments indicated sexual reproduction in A. elegans, asexuality in A. stylifer and A. karnyi, and both sexual and asexual lineages in A. rufus. Asexuality in A. stylifer and A. rufus coincides with a worldwide distribution, with sexual A. rufus lineages confined to a limited area. Inference of molecular phylogenies and antibiotic treatment revealed different causes of asexuality in different species. Asexuality in A. stylifer and A. karnyi has most likely genetic causes, while it is induced by endosymbionts in A. rufus. Endosymbiont-community characterization revealed presence of Wolbachia, and lack of other bacteria known to manipulate host reproduction. However, only 69% asexual A. rufus females are Wolbachia-infected, indicating that either an undescribed endosymbiont causes asexuality in this species or that Wolbachia was lost in several lineages that remained asexual. These results open new perspectives for studies on the maintenance of mixed sexual and asexual reproduction in natural populations.}, } @article {pmid24619863, year = {2015}, author = {Mathenge, CW and Riegler, M and Beattie, GA and Spooner-Hart, RN and Holford, P}, title = {Genetic variation amongst biotypes of Dactylopius tomentosus.}, journal = {Insect science}, volume = {22}, number = {3}, pages = {360-374}, doi = {10.1111/1744-7917.12120}, pmid = {24619863}, issn = {1744-7917}, mesh = {Animals ; Biological Control Agents ; Cactaceae/classification/*parasitology ; DNA, Bacterial/genetics ; Electron Transport Complex IV/genetics ; Genes, Mitochondrial ; Genetic Variation ; Hemiptera/*genetics/*microbiology ; Host-Parasite Interactions ; Phylogeny ; Wolbachia/genetics ; }, abstract = {The tomentose cochineal scale insect, Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae), is an important biological control agent against invasive species of Cylindropuntia (Caryophyllales: Cactaceae). Recent studies have demonstrated that this scale is composed of host-affiliated biotypes with differential host specificity and fitness on particular host species. We investigated genetic variation and phylogenetic relationships among D. tomentosus biotypes and provenances to examine the possibility that genetic diversity may be related to their host-use pattern, and whether their phylogenetic relationships would give insights into taxonomic relatedness of their host plants. Nucleotide sequence comparison was accomplished using sequences of the mitochondrial cytochrome c oxidase I (COI) gene. Sequences of individuals from the same host plant within a region were identical and characterized by a unique haplotype. Individuals belonging to the same biotype but from different regions had similar haplotypes. However, haplotypes were not shared between different biotypes. Phylogenetic analysis grouped the monophyletic D. tomentosus into 3 well-resolved clades of biotypes. The phylogenetic relationships and clustering of biotypes corresponded with known taxonomic relatedness of their hosts. Two biotypes, Fulgida and Mamillata, tested positive for Wolbachia (α-Proteobacteria), a common endosymbiont of insects. The Wolbachia sequences were serendipitously detected by using insect-specific COI DNA barcoding primers and are most similar to Wolbachia Supergroup F strains. This study is the first molecular characterization of cochineal biotypes that, together with Wolbachia sequences, contribute to the better identification of the biotypes of cochineal insects and to the biological control of cacti using host-specific biotypes of the scale.}, } @article {pmid24618033, year = {2015}, author = {Zug, R and Hammerstein, P}, title = {Bad guys turned nice? A critical assessment of Wolbachia mutualisms in arthropod hosts.}, journal = {Biological reviews of the Cambridge Philosophical Society}, volume = {90}, number = {1}, pages = {89-111}, doi = {10.1111/brv.12098}, pmid = {24618033}, issn = {1469-185X}, mesh = {Animals ; Arthropods/*microbiology ; Biological Evolution ; Host-Pathogen Interactions ; Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are the most abundant bacterial endosymbionts among arthropods. Although maternally inherited, they do not conform to the widespread view that vertical transmission inevitably selects for beneficial symbionts. Instead, Wolbachia are notorious for their reproductive parasitism which, although lowering host fitness, ensures their spread. However, even for reproductive parasites it can pay to enhance host fitness. Indeed, there is a recent upsurge of reports on Wolbachia-associated fitness benefits. Therefore, the question arises how such instances of mutualism are related to the phenotypes of reproductive parasitism. Here, we review the evidence of Wolbachia mutualisms in arthropods, including both facultative and obligate relationships, and critically assess their biological relevance. Although many studies report anti-pathogenic effects of Wolbachia, few actually prove these effects to be relevant to field conditions. We further show that Wolbachia frequently have beneficial and detrimental effects at the same time, and that reproductive manipulations and obligate mutualisms may share common mechanisms. These findings undermine the idea of a clear-cut distinction between Wolbachia mutualism and parasitism. In general, both facultative and obligate mutualisms can have a strong, and sometimes unforeseen, impact on the ecology and evolution of Wolbachia and their arthropod hosts. Acknowledging this mutualistic potential might be the key to a better understanding of some unresolved issues in the study of Wolbachia-host interactions.}, } @article {pmid24612968, year = {2014}, author = {Tochen, S and Dalton, DT and Wiman, N and Hamm, C and Shearer, PW and Walton, VM}, title = {Temperature-related development and population parameters for Drosophila suzukii (Diptera: Drosophilidae) on cherry and blueberry.}, journal = {Environmental entomology}, volume = {43}, number = {2}, pages = {501-510}, doi = {10.1603/EN13200}, pmid = {24612968}, issn = {1938-2936}, mesh = {Agriculture ; Analysis of Variance ; Animals ; Blueberry Plants/*parasitology ; Drosophila/*growth & development ; Female ; Fertility/physiology ; Longevity/physiology ; Models, Biological ; Oregon ; Prunus/*parasitology ; Statistics, Nonparametric ; *Temperature ; }, abstract = {Temperature-related studies were conducted on Drosophila suzukii Matsumura (Diptera: Drosophilidae: Drosophilini). From 10-28°C, temperature had a significant impact on blueberries, Vaccinium corymbosum L. (Ericales: Ericaceae), and cherries, Prunus avium (L.) L. 1755 (Rosales: Rosaceae), important commercial hosts of D. suzukii. Temperature had a significant influence on D. suzukii developmental period, survival, and fecundity, with decreasing developmental periods as temperatures increased to 28°C. At 30°C, the highest temperature tested, development periods increased, indicating that above this temperature the developmental extremes for the species were approached. D. suzukii reared on blueberries had lower fecundity than reared on cherries at all temperatures where reproduction occurred. The highest net reproductive rate (R(o)) and intrinsic rate of population increase (r(m)) were recorded on cherries at 22°C and was 195.1 and 0.22, respectively. Estimations using linear and nonlinear fit for the minimum, optimal, and maximum temperatures where development can take place were respectively, 7.2, 28.1, and 42.1°C. The r(m) values were minimal, optimal, and maximal at 13.4, 21.0, and 29.3°C, respectively. Our laboratory cultures of D. suzukii displayed high rates of infection for Wolbachia spp. (Rickettsiales: Rickettsiaceae), and this infection may have impacted fecundity found in this study. A temperature-dependent matrix population estimation model using fecundity and survival data were run to determine whether these data could predict D. suzukii pressure based on environmental conditions. The model was applied to compare the 2011 and 2012 crop seasons in an important cherry production region. Population estimates using the model explained different risk levels during the key cherry harvest period between these seasons.}, } @article {pmid24612104, year = {2014}, author = {Lewis, SE and Rice, A and Hurst, GD and Baylis, M}, title = {First detection of endosymbiotic bacteria in biting midges Culicoides pulicaris and Culicoides punctatus, important Palaearctic vectors of bluetongue virus.}, journal = {Medical and veterinary entomology}, volume = {28}, number = {4}, pages = {453-456}, doi = {10.1111/mve.12055}, pmid = {24612104}, issn = {1365-2915}, mesh = {Animals ; Bacteroidaceae/*genetics/*isolation & purification ; Bluetongue virus/*physiology ; Ceratopogonidae/*microbiology ; Female ; Insect Vectors/*microbiology/*virology ; Male ; Phylogeny ; Species Specificity ; Symbiosis ; }, abstract = {Heritable bacteria have been highlighted as important components of vector biology, acting as required symbionts with an anabolic role, altering competence for disease transmission, and affecting patterns of gene flow by altering cross compatibility. In this paper, we tested eight U.K. species of Culicoides (Diptera: Ceratopogonidae) midge for the presence of five genera of endosymbiotic bacteria: Cardinium (Bacteroidales: Bacteroidaceae); Wolbachia (Rickettsiales: Rickettsiaceae); Spiroplasma (Entomoplasmatales: Spiroplasmataceae); Arsenophonus (Enterobacteriales: Enterobacteriaceae), and Rickettsia (Rickettsiales: Rickettsiaceae). Cardinium spp. were detected in both sexes of Culicoides pulicaris and Culicoides punctatus, two known vectors of bluetongue virus. Cardinium spp. were not detected in any other species, including the Culicoides obsoletus group, the main vector of bluetongue and Schmallenberg viruses in northern Europe. The other endosymbionts were not detected in any Culicoides species. The Cardinium strain detected in U.K. Culicoides species is very closely related to the Candidatus Cardinium hertigii group C, previously identified in Culicoides species in Asia. Further, we infer that the symbiont is not a sex ratio distorter and shows geographic variation in prevalence within a species. Despite its detection in several species of Culicoides that vector arboviruses worldwide, the absence of Cardinium in the C. obsoletus group suggests that infections of these symbionts may not be necessary to the arboviral vector competence of biting midges.}, } @article {pmid24605471, year = {2014}, author = {Ritchie, SA and Buhagiar, TS and Townsend, M and Hoffmann, A and Van Den Hurk, AF and McMahon, JL and Eiras, AE}, title = {Field validation of the gravid Aedes trap (GAT) for collection of Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {51}, number = {1}, pages = {210-219}, doi = {10.1603/me13105}, pmid = {24605471}, issn = {0022-2585}, mesh = {*Aedes/virology ; Animals ; Dengue Virus/isolation & purification ; Female ; Insect Vectors/virology ; Mosquito Control/*instrumentation ; RNA, Viral/*analysis ; }, abstract = {Current surveillance methods for adult Aedes aegypti (L.) are expensive, require electrical power (e.g., the BG-Sentinel trap, BGS), are labor intensive (aspirators), or require difficult to use and costly adhesives (sticky ovitraps). Field trials were conducted in Cairns (Australia) to compare the efficacy of the newly designed Gravid Aedes Trap (GAT) against existing sticky ovitraps (MosquiTRAP and double sticky ovitrap) and the BGS. Latin square design trials confirmed that alarge GAT using a 9.2-liters bucket treated with Mortein Barrier Outdoor Surface Spray ([AI] 0.3 g/kg imiprothrin and 0.6 g/kg deltamethrin) outperformed a smaller 1.2-liters GAT and collected, on average, 3.7x and 2.4X more female Ae. aegypti than the MosquiTRAP and double sticky ovitrap, respectively. Field trials showed that the GAT collected 10-50% less female Ae. aegypti than the BGS trap but 30% more gravid mosquitoes than the BGS. Trials using the BGS and the GAT indicated that there was no difference in capture rates between female Ae. aegypti uninfected and infected with the wMel strain of Wolbachia, and wMel infection rates were nearly identical at >90% to field captured Ae. aegypti. The potential for the GAT to be used for dengue virus surveillance was also demonstrated with dengue virus type 3 RNA detected in five-sixths and six-sixths pools ofAe. aegypti stored in a GAT held at 28 degreeC and 60% relative humidity for 7 and 14 d, respectively. Mosquito knock down in GATs treated with Mortein surface spray set in 30, 70, and 99% shade was comparable for up to 2 mo, with only approximately 10% of adults escaping. The GAT is therefore a useful tool for capturing adult Ae. aegypti and may be suitable for other container-inhabiting species such as Aedes albopictus (Skuse) and Culex quinquefasciatus Say. The low cost and practicality of operation make the GAT suitable for vector surveillance and projects requiring monitoring of mosquitoes for Wolbachia and arboviruses, especially in developing countries.}, } @article {pmid24605469, year = {2014}, author = {Micieli, MV and Glaser, RL}, title = {Somatic Wolbachia (Rickettsiales: Rickettsiaceae) levels in Culex quinquefasciatus and Culex pipiens (Diptera: Culicidae) and resistance to West Nile virus infection.}, journal = {Journal of medical entomology}, volume = {51}, number = {1}, pages = {189-199}, doi = {10.1603/me13152}, pmid = {24605469}, issn = {0022-2585}, support = {AI076258/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culex/immunology/*microbiology ; Female ; Insect Vectors/immunology/*microbiology ; *West Nile virus ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacteria Wolbachia pipientis Hertig infects a wide variety of insect species and can increase viral resistance in its host. Wolbachia naturally infects Culex quinquefasciatus Say and Culex pipiens L. mosquitoes, both vectors of West Nile virus (WNV). We recently demonstrated that Wolbachia infection of Cx. quinquefasciatus laboratory strain Ben95 increases host resistance to WNV infection, reducing vector competence. This observation raised the possibility that Wolbachia could impact vector competence in other populations of Cx. quinquefasciatus or Cx. pipiens. To investigate this possibility, Wolbachia densities were measured in Ben95 Cx. quinquefasciatus and compared with densities in a newly established colony of Cx. quinquefasciatus, and in field-collected and colonized Cx. pipiens. Wolbachia densities in somatic tissues of Ben95 Cx. quinquefasciatus were significantly higher than densities in the other mosquito populations tested. There was also no significant spatiotemporal variation in Wolbachia density in the field-collected Cx. pipiens, although significant familial differences were observed. Correlating Wolbachia densities and vector competence in individual colonized Cx. pipiens indicated that the densities of somatic Wolbachia observed in the mosquitoes other than Ben95 Cx. quinquefasciatus were too low to inhibit WNV infection and reduce vector competence. These results suggest that the high Wolbachia densities capable of inducing resistance to WNV in Ben95 Cx. quinquefasciatus are not a general characteristic of Cx. quinquefasciatus or Cx. pipiens mosquitoes and that the impact of Wolbachia on vector competence in field populations of Cx. quinquefasciatus and Cx. pipiens, if any, is likely to be limited to specific populations.}, } @article {pmid24600010, year = {2014}, author = {Kliot, A and Cilia, M and Czosnek, H and Ghanim, M}, title = {Implication of the bacterial endosymbiont Rickettsia spp. in interactions of the whitefly Bemisia tabaci with tomato yellow leaf curl virus.}, journal = {Journal of virology}, volume = {88}, number = {10}, pages = {5652-5660}, pmid = {24600010}, issn = {1098-5514}, mesh = {Animals ; Begomovirus/*isolation & purification ; Carrier State/*virology ; Gastrointestinal Tract/microbiology/virology ; Hemiptera/*microbiology/*virology ; Microbial Interactions ; Rickettsia/*isolation & purification/*physiology ; *Symbiosis ; }, abstract = {UNLABELLED: Numerous animal and plant viruses are transmitted by arthropod vectors in a persistent, circulative manner. Tomato yellow leaf curl virus (TYLCV) is transmitted by the sweet potato whitefly Bemisia tabaci. We report here that infection with Rickettsia spp., a facultative endosymbiont of whiteflies, altered TYLCV-B. tabaci interactions. A B. tabaci strain infected with Rickettsia acquired more TYLCV from infected plants, retained the virus longer, and exhibited nearly double the transmission efficiency compared to an uninfected B. tabaci strain with the same genetic background. Temporal and spatial antagonistic relationships were discovered between Rickettsia and TYLCV within the whitefly. In different time course experiments, the levels of virus and Rickettsia within the insect were inversely correlated. Fluorescence in situ hybridization analysis of Rickettsia-infected midguts provided evidence for niche exclusion between Rickettsia and TYLCV. In particular, high levels of the bacterium in the midgut resulted in higher virus concentrations in the filter chamber, a favored site for virus translocation along the transmission pathway, whereas low levels of Rickettsia in the midgut resulted in an even distribution of the virus. Taken together, these results indicate that Rickettsia, by infecting the midgut, increases TYLCV transmission efficacy, adding further insights into the complex association between persistent plant viruses, their insect vectors, and microorganism tenants that reside within these insects.

IMPORTANCE: Interest in bacterial endosymbionts in arthropods and many aspects of their host biology in agricultural and human health systems has been increasing. A recent and relevant studied example is the influence of Wolbachia on dengue virus transmission by mosquitoes. In parallel with our recently studied whitefly-Rickettsia-TYLCV system, other studies have shown that dengue virus levels in the mosquito vector are inversely correlated with bacterial load. Our work here presents evidence of unifying principles between vectors of plant and animal viruses in a role for endosymbionts in manipulating vector biology and pathogen transmission. Our results demonstrate the influence of an interesting and prominent bacterial endosymbiont in Bemisia tabaci in TYLCV transmission, a worldwide disease infecting tomatoes. Besides its agricultural importance, this system provides interesting insights into Bemisia interaction with these newly discovered endosymbionts.}, } @article {pmid24598598, year = {2014}, author = {Taylor, CM and Coffey, PL and DeLay, BD and Dively, GP}, title = {The importance of gut symbionts in the development of the brown marmorated stink bug, Halyomorpha halys (Stål).}, journal = {PloS one}, volume = {9}, number = {3}, pages = {e90312}, pmid = {24598598}, issn = {1932-6203}, mesh = {Animals ; DNA, Bacterial/genetics ; Digestive System/*microbiology ; Female ; Fertility ; Heteroptera/*microbiology/physiology ; Male ; Movement ; Nymph/microbiology/physiology ; Ovum/microbiology ; Pantoea/genetics ; Symbiosis ; Wolbachia/genetics ; }, abstract = {The invasive brown marmorated stink bug, Halyomorpha halys (Stål), has become a severe agricultural pest and nuisance problem since its introduction in the U.S. Research is being conducted to understand its biology and to find management solutions. Its symbiotic relationship with gut symbionts is one aspect of its biology that is not understood. In the family Pentatomidae, the reliance on gut symbionts for successful development seems to vary depending on the species of stink bug. This research assessed the role of gut symbionts in the development, survivorship, and fecundity of H. halys. We compared various fitness parameters of nymphs and adults reared from surface sterilized and untreated egg masses during two consecutive generations under laboratory conditions. Results provided direct evidence that H. halys is negatively impacted by the prevention of vertical transmission of its gut symbionts and that this impact is significant in the first generation and manifests dramatically in the subsequent generation. Developmental time and survivorship of treated cohorts in the first generation were significantly affected during third instar development through to the adult stage. Adults from the sterilized treatment group exhibited longer pre-oviposition periods, produced fewer egg masses, had significantly smaller clutch sizes, and the hatch rate and survivorship of those eggs were significantly reduced. Observations following hatch of surface sterilized eggs also revealed significant effects on wandering behavior of the first instars. The second generation progeny from adults of the sterilized cohorts showed significantly lower survival to adulthood, averaging only 0.3% compared to 20.8% for the control cohorts. Taken together, results demonstrate that H. halys is heavily impacted by deprival of its gut symbionts. Given the economic status of this invasive pest, further investigations may lead to management tactics that disrupt this close symbiotic relationship in the biology of H. halys.}, } @article {pmid24596970, year = {2014}, author = {Andresh, J}, title = {[Can genetically modified mosquitos help in the battle against viral diseases?].}, journal = {Deutsche medizinische Wochenschrift (1946)}, volume = {139}, number = {5}, pages = {176-177}, doi = {10.1055/s-0033-1353965}, pmid = {24596970}, issn = {1439-4413}, mesh = {Animals ; Animals, Genetically Modified/*genetics/immunology ; Cross Protection/genetics/immunology ; Culicidae/*genetics/immunology ; Dengue/immunology/*prevention & control/*transmission ; Gene Transfer Techniques ; Humans ; Population Dynamics ; Repetitive Sequences, Nucleic Acid/genetics ; Serotyping ; Wolbachia/genetics ; Yellow Fever/immunology/*prevention & control/*transmission ; }, } @article {pmid24596263, year = {2014}, author = {Bansal, R and Mian, MA and Michel, AP}, title = {Microbiome diversity of Aphis glycines with extensive superinfection in native and invasive populations.}, journal = {Environmental microbiology reports}, volume = {6}, number = {1}, pages = {57-69}, doi = {10.1111/1758-2229.12108}, pmid = {24596263}, issn = {1758-2229}, mesh = {Animals ; Aphids/*microbiology ; Bacteria/classification/genetics/*isolation & purification ; *Biodiversity ; Introduced Species ; Japan ; *Microbiota ; North America ; Phylogeny ; Republic of Korea ; }, abstract = {Associations among insects and microbes can lead to beneficial or parasitic interactions. Using 454 sequencing of 16S RNA genes, we compared microbiome diversity and abundance among field-collected (F) and laboratory-reared (L) populations of the invasive soybean aphid (Aphis glycines), a pest of soybean. Additionally, we screened A. glycines populations from native (Japan, South Korea and China) and invasive regions (North America) to broadly determine the microbiome diversity. Our results suggested that Arsenophonus (relative abundance of 54.6%), Buchnera (38.7%) and Wolbachia (3.7%) were the major bacteria associated with A. glycines. Arsenophonus was the most abundant in F populations but was significantly reduced in L populations; additional bacteria species also had lower relative abundances in L populations. Native and invasive populations were largely similar in bacteria communities and revealed substantial superinfection of Arsenophonus and Wolbachia. The lone exception was a lack of Arsenophonus in A. glycines from Japan. Divergent selection pressures among natural and laboratory populations were inferred as factors driving the differential bacterial communities observed. Our results will allow for improved comparative aphid-symbiont research and broaden our understanding of the interactions among insects, endosymbionts and their environments.}, } @article {pmid24594301, year = {2014}, author = {Correa, CC and Ballard, JW}, title = {What can symbiont titres tell us about co-evolution of Wolbachia and their host?.}, journal = {Journal of invertebrate pathology}, volume = {118}, number = {}, pages = {20-27}, doi = {10.1016/j.jip.2014.02.009}, pmid = {24594301}, issn = {1096-0805}, mesh = {Adaptation, Physiological/*genetics ; Animals ; *Biological Evolution ; Drosophila/*microbiology ; Female ; Gonads/microbiology ; Infectious Disease Transmission, Vertical ; Male ; Symbiosis/*genetics ; Wolbachia/*genetics ; }, abstract = {There is a long-standing prediction that associations with vertically transmitted symbionts evolve towards maximisation of host reproductive success, eventually leading to mutualist symbiosis and coadaptation. Under this scenario, the regulation of symbiont titres in host tissues would be expected to be greater when partners have coevolved for a long time than when they have recently met. Wolbachia pipientis, a common vertically transmitted symbiont of invertebrates, often has the capacity to spread through the host population without being beneficial to the hosts, by means of reducing the hatch rate in crosses between uninfected females and infected males. This manipulation, namely cytoplasmic incompatibility (CI), may exert strong selection on the accuracy of infection transmission from mother to offspring, and therefore, on regulation of symbiont titres in the ova. Here, we examined the symbiont density dynamics in gonads of Drosophila simulans infected with the wMa strain of Wolbachia, known to cause mild CI and likely to be the oldest Wolbachia infection known to this fly species. Further, we compared these results with those obtained for the more recent association between D. simulans and the potent CI-inducer wHa (Correa and Ballard, 2012). We aimed to determine if the regulation of Wolbachia density in fly gonads is greater in the older association, as would be predicted solely by gradual coadaptation, or if the selection exerted by CI on reproductive fitness could also play a role, therefore showing tighter regulation on flies with the stronger CI-inducing strain. We observed that Wolbachia density in gonads of wMa infected flies changed with laboratory adaptation and were disturbed by environmental challenges, which contrasted with the stability of ovarian wHa density to the same treatments. Our observations are in line with the prediction that selection on reproductive fitness influences the evolution symbiont density regulation in Drosophila, and may provide insights into the evolutionary processes involved in the maintenance or loss of Wolbachia.}, } @article {pmid24587459, year = {2014}, author = {Frentiu, FD and Zakir, T and Walker, T and Popovici, J and Pyke, AT and van den Hurk, A and McGraw, EA and O'Neill, SL}, title = {Limited dengue virus replication in field-collected Aedes aegypti mosquitoes infected with Wolbachia.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {2}, pages = {e2688}, pmid = {24587459}, issn = {1935-2735}, mesh = {Aedes/microbiology/*virology ; Animals ; Dengue Virus/genetics/*isolation & purification/physiology ; Female ; Insect Vectors/microbiology/*virology ; Pest Control, Biological/*methods ; RNA, Viral/analysis/genetics ; Virus Replication/*physiology ; *Wolbachia ; }, abstract = {INTRODUCTION: Dengue is one of the most widespread mosquito-borne diseases in the world. The causative agent, dengue virus (DENV), is primarily transmitted by the mosquito Aedes aegypti, a species that has proved difficult to control using conventional methods. The discovery that A. aegypti transinfected with the wMel strain of Wolbachia showed limited DENV replication led to trial field releases of these mosquitoes in Cairns, Australia as a biocontrol strategy for the virus.

Field collected wMel mosquitoes that were challenged with three DENV serotypes displayed limited rates of body infection, viral replication and dissemination to the head compared to uninfected controls. Rates of dengue infection, replication and dissemination in field wMel mosquitoes were similar to those observed in the original transinfected wMel line that had been maintained in the laboratory. We found that wMel was distributed in similar body tissues in field mosquitoes as in laboratory ones, but, at seven days following blood-feeding, wMel densities increased to a greater extent in field mosquitoes.

CONCLUSIONS/SIGNIFICANCE: Our results indicate that virus-blocking is likely to persist in Wolbachia-infected mosquitoes after their release and establishment in wild populations, suggesting that Wolbachia biocontrol may be a successful strategy for reducing dengue transmission in the field.}, } @article {pmid24579829, year = {2014}, author = {McFrederick, QS and Wcislo, WT and Hout, MC and Mueller, UG}, title = {Host species and developmental stage, but not host social structure, affects bacterial community structure in socially polymorphic bees.}, journal = {FEMS microbiology ecology}, volume = {88}, number = {2}, pages = {398-406}, doi = {10.1111/1574-6941.12302}, pmid = {24579829}, issn = {1574-6941}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; Bees/growth & development/*microbiology ; Biodiversity ; Lactobacillus/genetics/isolation & purification ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Social Behavior ; Wolbachia/genetics/isolation & purification ; }, abstract = {Social transmission and host developmental stage are thought to profoundly affect the structure of bacterial communities associated with honey bees and bumble bees, but these ideas have not been explored in other bee species. The halictid bees Megalopta centralis and M. genalis exhibit intrapopulation social polymorphism, which we exploit to test whether bacterial communities differ by host social structure, developmental stage, or host species. We collected social and solitary Megalopta nests and sampled bees and nest contents from all stages of host development. To survey these bacterial communities, we used 16S rRNA gene 454 pyrosequencing. We found no effect of social structure, but found differences by host species and developmental stage. Wolbachia prevalence differed between the two host species. Bacterial communities associated with different developmental stages appeared to be driven by environmentally acquired bacteria. A Lactobacillus kunkeei clade bacterium that is consistently associated with other bee species was dominant in pollen provisions and larval samples, but less abundant in mature larvae and pupae. Foraging adults appeared to often reacquire L. kunkeei clade bacteria, likely while foraging at flowers. Environmental transmission appears to be more important than social transmission for Megalopta bees at the cusp between social and solitary behavior.}, } @article {pmid24568185, year = {2014}, author = {Lentz, CS and Halls, VS and Hannam, JS and Strassel, S and Lawrence, SH and Jaffe, EK and Famulok, M and Hoerauf, A and Pfarr, KM}, title = {wALADin benzimidazoles differentially modulate the function of porphobilinogen synthase orthologs.}, journal = {Journal of medicinal chemistry}, volume = {57}, number = {6}, pages = {2498-2510}, pmid = {24568185}, issn = {1520-4804}, support = {R01 ES003654/ES/NIEHS NIH HHS/United States ; R56 AI077577/AI/NIAID NIH HHS/United States ; AI077577/AI/NIAID NIH HHS/United States ; ES003654/ES/NIEHS NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/*chemical synthesis/*pharmacology ; Antiprotozoal Agents/chemical synthesis/pharmacology ; Benzimidazoles/*chemical synthesis/*pharmacology ; Chlamydia/drug effects ; Herbicides/chemical synthesis/pharmacology ; Humans ; Hydrogen-Ion Concentration ; Microbial Sensitivity Tests ; Molecular Weight ; Pisum sativum ; Plants ; Porphobilinogen Synthase/*antagonists & inhibitors ; Porphyrias/drug therapy ; Pseudomonas aeruginosa/drug effects/enzymology ; Rickettsia/drug effects ; Stereoisomerism ; Structure-Activity Relationship ; Wolbachia/drug effects ; }, abstract = {The heme biosynthesis enzyme porphobilinogen synthase (PBGS) is a potential drug target in several human pathogens. wALADin1 benzimidazoles have emerged as species-selective PBGS inhibitors against Wolbachia endobacteria of filarial worms. In the present study, we have systematically tested wALADins against PBGS orthologs from bacteria, protozoa, metazoa, and plants to elucidate the inhibitory spectrum. However, the effect of wALADin1 on different PBGS orthologs was not limited to inhibition: several orthologs were stimulated by wALADin1; others remained unaffected. We demonstrate that wALADins allosterically modulate the PBGS homooligomeric equilibrium with inhibition mediated by favoring low-activity oligomers, while 5-aminolevulinic acid, Mg(2+), or K(+) stabilized high-activity oligomers. Pseudomonas aeruginosa PBGS could be inhibited or stimulated by wALADin1 depending on these factors and pH. We have defined the wALADin chemotypes responsible for either inhibition or stimulation, facilitating the design of tailored PBGS modulators for potential application as antimicrobial agents, herbicides, or drugs for porphyric disorders.}, } @article {pmid24561051, year = {2014}, author = {Cerveau, N and Bouchon, D and Bergès, T and Grève, P}, title = {Molecular evolution of the androgenic hormone in terrestrial isopods.}, journal = {Gene}, volume = {540}, number = {1}, pages = {71-77}, doi = {10.1016/j.gene.2014.02.024}, pmid = {24561051}, issn = {1879-0038}, mesh = {Amino Acid Sequence ; Animals ; Arthropod Proteins/chemistry/*genetics ; *Evolution, Molecular ; Gonadal Hormones/chemistry/*genetics ; Isopoda/*genetics ; Male ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, Protein ; Sex Differentiation/genetics ; }, abstract = {In crustaceans, the androgenic gland (AG), thanks to the synthesis of the androgenic gland hormone (AGH), controls the differentiation of the primary and secondary male sexual characters. In this study, we amplified 12 new AGH cDNAs in species belonging to five different families of the infra-order Ligiamorpha of terrestrial isopods. Putative essential amino acids for the production of a functional AGH protein exhibit signatures of negative selection and are strictly conserved including typical proteolytic cleavage motifs, a putative N-linked glycosylation motif on the A chains and the eight Cys positions. An insulin-like growth factor motif was also identified in Armadillidium AGH sequences. The phylogenetic relationships of AGH sequences allowed one to distinguish two main clades, corresponding to members of the Armadillidiidae and the Porcellionidae families which are congruent with the narrow specificity of AG heterospecific grafting. An in-depth understanding of the regulation of AGH expression would help deciphering the interaction between Wolbachia, widespread feminizing endosymbiotic bacteria in isopods, and the sex differentiation of their hosts.}, } @article {pmid24551251, year = {2014}, author = {Hugo, LE and Jeffery, JA and Trewin, BJ and Wockner, LF and Nguyen, TY and Nguyen, HL and Nghia, le T and Hine, E and Ryan, PA and Kay, BH}, title = {Adult survivorship of the dengue mosquito Aedes aegypti varies seasonally in central Vietnam.}, journal = {PLoS neglected tropical diseases}, volume = {8}, number = {2}, pages = {e2669}, pmid = {24551251}, issn = {1935-2735}, mesh = {*Aedes/microbiology/virology ; Animals ; *Dengue/prevention & control/transmission ; Dengue Virus/genetics/*isolation & purification ; *Insect Vectors/microbiology/virology ; Seasons ; Vietnam ; Wolbachia ; }, abstract = {The survival characteristics of the mosquito Aedes aegypti affect transmission rates of dengue because transmission requires infected mosquitoes to survive long enough for the virus to infect the salivary glands. Mosquito survival is assumed to be high in tropical, dengue endemic, countries like Vietnam. However, the survival rates of wild populations of mosquitoes are seldom measured due the difficulty of predicting mosquito age. Hon Mieu Island in central Vietnam is the site of a pilot release of Ae. aegypti infected with a strain of Wolbachia pipientis bacteria (wMelPop) that induces virus interference and mosquito life-shortening. We used the most accurate mosquito age grading approach, transcriptional profiling, to establish the survival patterns of the mosquito population from the population age structure. Furthermore, estimations were validated on mosquitoes released into a large semi-field environment consisting of an enclosed house, garden and yard to incorporate natural environmental variability. Mosquito survival was highest during the dry/cool (January-April) and dry/hot (May-August) seasons, when 92 and 64% of Hon Mieu mosquitoes had survived to an age that they were able to transmit dengue (12 d), respectively. This was reduced to 29% during the wet/cool season from September to December. The presence of Ae. aegypti older than 12 d during each season is likely to facilitate the observed continuity of dengue transmission in the region. We provide season specific Ae. aegypti survival models for improved dengue epidemiology and evaluation of mosquito control strategies that aim to reduce mosquito survival to break the dengue transmission cycle.}, } @article {pmid24551238, year = {2014}, author = {Ruther, J and McCaw, J and Böcher, L and Pothmann, D and Putz, I}, title = {Pheromone diversification and age-dependent behavioural plasticity decrease interspecific mating costs in Nasonia.}, journal = {PloS one}, volume = {9}, number = {2}, pages = {e89214}, pmid = {24551238}, issn = {1932-6203}, mesh = {Aging/drug effects/*physiology ; Animals ; Biological Assay ; Female ; Male ; Mating Preference, Animal/*drug effects ; Models, Biological ; Olfactometry ; Sex Attractants/*pharmacology ; Sex Ratio ; Sexual Behavior, Animal/*drug effects ; Species Specificity ; Wasps/drug effects/*physiology ; }, abstract = {Interspecific mating can cause severe fitness costs due to the fact that hybrids are often non-viable or less fit. Thus, theory predicts the selection of traits that lessen reproductive interactions between closely related sympatric species. Males of the parasitic wasp Nasonia vitripennis differ from all other Nasonia species by an additional sex pheromone component, but the ecological selective forces underlying this pheromone diversification are unknown. Here we present data from lab experiments suggesting that costly interspecific sexual interactions with the sympatric species N. giraulti might have been responsible for the pheromone evolution and some courtship-related behavioural adaptations in N. vitripennis. Most N. giraulti females are inseminated already within the host, but N. giraulti males still invest in costly sex pheromones after emergence. Furthermore, they do not discriminate between N. vitripennis females and conspecifics during courtship. Therefore, N. vitripennis females, most of which emerge as virgins, face the risk of mating with N. giraulti resulting in costly all-male broods due to Wolbachia-induced cytoplasmic incompatibility. As a counter adaptation, young N. vitripennis females discriminate against N. giraulti males using the more complex conspecific sex pheromone and reject most of them during courtship. With increasing age, however, N. vitripennis females become less choosy, but often compensate mating errors by re-mating with a conspecific. By doing so, they can principally avoid suboptimal offspring sex ratios, but a microcosm experiment suggests that under more natural conditions N. vitripennis females cannot completely avoid fitness costs due to heterospecific mating. Our study provides support for the hypothesis that communication interference of closely related sympatric species using similar sexual signals can generate selective pressures that lead to their divergence.}, } @article {pmid24533099, year = {2014}, author = {Yu, H and Ji, R and Ye, W and Chen, H and Lai, W and Fu, Q and Lou, Y}, title = {Transcriptome analysis of fat bodies from two brown planthopper (Nilaparvata lugens) populations with different virulence levels in rice.}, journal = {PloS one}, volume = {9}, number = {2}, pages = {e88528}, pmid = {24533099}, issn = {1932-6203}, mesh = {Animals ; Carbohydrates/chemistry ; DNA, Complementary/metabolism ; Fat Body/*physiology ; Female ; Gene Expression Profiling ; Gene Library ; Genome ; Hemiptera/*genetics/*microbiology ; Lipid Metabolism ; Oryza ; RNA, Messenger/metabolism ; Signal Transduction ; Species Specificity ; *Transcriptome ; Virulence ; Wolbachia/genetics ; Xenobiotics/chemistry ; }, abstract = {BACKGROUND: The brown planthopper (BPH), Nilaparvata lugens (Stål), one of the most serious rice insect pests in Asia, can quickly overcome rice resistance by evolving new virulent populations. The insect fat body plays essential roles in the life cycles of insects and in plant-insect interactions. However, whether differences in fat body transcriptomes exist between insect populations with different virulence levels and whether the transcriptomic differences are related to insect virulence remain largely unknown.

In this study, we performed transcriptome-wide analyses on the fat bodies of two BPH populations with different virulence levels in rice. The populations were derived from rice variety TN1 (TN1 population) and Mudgo (M population). In total, 33,776 and 32,332 unigenes from the fat bodies of TN1 and M populations, respectively, were generated using Illumina technology. Gene ontology annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology classifications indicated that genes related to metabolism and immunity were significantly active in the fat bodies. In addition, a total of 339 unigenes showed homology to genes of yeast-like symbionts (YLSs) from 12 genera and endosymbiotic bacteria Wolbachia. A comparative analysis of the two transcriptomes generated 7,860 differentially expressed genes. GO annotations and enrichment analysis of KEGG pathways indicated these differentially expressed transcripts might be involved in metabolism and immunity. Finally, 105 differentially expressed genes from YLSs and Wolbachia were identified, genes which might be associated with the formation of different virulent populations.

CONCLUSIONS/SIGNIFICANCE: This study was the first to compare the fat-body transcriptomes of two BPH populations having different virulence traits and to find genes that may be related to this difference. Our findings provide a molecular resource for future investigations of fat bodies and will be useful in examining the interactions between the fat body and virulence variation in the BPH.}, } @article {pmid24517908, year = {2014}, author = {Arp, A and Munyaneza, JE and Crosslin, JM and Trumble, J and Bextine, B}, title = {A global comparison of Bactericera cockerelli (Hemiptera: Triozidae) microbial communities.}, journal = {Environmental entomology}, volume = {43}, number = {2}, pages = {344-352}, doi = {10.1603/EN13256}, pmid = {24517908}, issn = {1938-2936}, mesh = {Animal Distribution/*physiology ; Animals ; Base Sequence ; *Biota ; Haplotypes ; Hemiptera/genetics/*microbiology ; Molecular Sequence Data ; New Zealand ; Nicaragua ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; United States ; }, abstract = {The potato psyllid (Bactericera cockerelli Sulc) is an economically important insect pest of solanaceous crops such as potato, tomato, pepper, and tobacco. Historically, the potato psyllid's range included central United States, Mexico, and California; more recently, populations of this insect have been reported in Central America, the Pacific Northwest, and New Zealand. Like most phytophagous insects, potato psyllids require symbiotic bacteria to compensate for nutritional deficiencies in their diet. Potato psyllids harbor the primary symbiont, Candidatus Carsonella ruddii, and may also harbor many secondary symbionts such as Wolbachia sp., Sodalis sp., Pseudomonas sp., and others. These secondary symbionts can have an effect on reproduction, nutrition, immune response, and resistances to heat or pesticides. To identify regional differences in potato psyllid bacterial symbionts, 454 pyrosequencing was performed using generic 16S rRNA gene primers. Analysis was performed using the Qiime 1.6.0 software suite, ARB Silva, and R. Operational taxonomic units were then grouped at 97% identity. Representative sequences were classified to genus using the ARB SILVA database. Potato psyllids collected in California contained a less diverse microbial community than those collected in the central United States and Central America. The crop variety, collection year, and haplotype did not seem to affect the microbial community in potato psyllids. The primary difference between psyllids in different regions was the presence and overall bacterial community composition of Candidatus Carsonella ruddii and Wolbachia.}, } @article {pmid24514880, year = {2014}, author = {Ribeiro, JMC and Chagas, AC and Pham, VM and Lounibos, LP and Calvo, E}, title = {An insight into the sialome of the frog biting fly, Corethrella appendiculata.}, journal = {Insect biochemistry and molecular biology}, volume = {44}, number = {}, pages = {23-32}, pmid = {24514880}, issn = {1879-0240}, support = {R01 AI044793/AI/NIAID NIH HHS/United States ; R21 AI095780/AI/NIAID NIH HHS/United States ; ZIA AI000810-17//Intramural NIH HHS/United States ; }, abstract = {The Nematocera infraorder Culicomorpha is believed to have descended from bloodfeeding ancestors over 200 million years ago, generating bloodfeeding and non-bloodfeeding flies in two superfamilies, the Culicoidea-containing the mosquitoes, the frog-feeding midges, the Chaoboridae, and the Dixidae-and the Chironomoidea-containing the black flies, the ceratopogonids, the Chironomidae, and the Thaumaleidae. Blood feeding requires many adaptations, including development of a sophisticated salivary potion that disarms host hemostasis, the physiologic mechanism comprising platelet aggregation, vasoconstriction, and blood clotting. The composition of the sialome (from the Greek sialo = saliva) from bloodfeeding animals can be inferred from analysis of their salivary gland transcriptome. While members of the mosquitoes, black flies, and biting midges have provided sialotranscriptome descriptions, no species of the frog-biting midges has been thus analyzed. We describe in this work the sialotranscriptome of Corethrella appendiculata, revealing a complex potion of enzymes, classical nematoceran protein families involved in blood feeding, and novel protein families unique to this species of frog-feeding fly. Bacterial (Wolbachia) and novel viral sequences were also discovered.}, } @article {pmid24500167, year = {2014}, author = {Zélé, F and Nicot, A and Berthomieu, A and Weill, M and Duron, O and Rivero, A}, title = {Wolbachia increases susceptibility to Plasmodium infection in a natural system.}, journal = {Proceedings. Biological sciences}, volume = {281}, number = {1779}, pages = {20132837}, pmid = {24500167}, issn = {1471-2954}, mesh = {Animals ; Culex/microbiology/*parasitology ; *Disease Resistance ; *Host-Parasite Interactions ; Plasmodium/pathogenicity/*physiology ; Wolbachia/*physiology ; }, abstract = {Current views about the impact of Wolbachia on Plasmodium infections are almost entirely based on data regarding artificially transfected mosquitoes. This work has shown that Wolbachia reduces the intensity of Plasmodium infections in mosquitoes, raising the exciting possibility of using Wolbachia to control or limit the spread of malaria. Whether natural Wolbachia infections have the same parasite-inhibiting properties is not yet clear. Wolbachia-mosquito combinations with a long evolutionary history are, however, key for understanding what may happen with Wolbachia-transfected mosquitoes after several generations of coevolution. We investigate this issue using an entirely natural mosquito-Wolbachia-Plasmodium combination. In contrast to most previous studies, which have been centred on the quantification of the midgut stages of Plasmodium, we obtain a measurement of parasitaemia that relates directly to transmission by following infections to the salivary gland stages. We show that Wolbachia increases the susceptibility of Culex pipiens mosquitoes to Plasmodium relictum, significantly increasing the prevalence of salivary gland stage infections. This effect is independent of the density of Wolbachia in the mosquito. These results suggest that naturally Wolbachia-infected mosquitoes may, in fact, be better vectors of malaria than Wolbachia-free ones.}, } @article {pmid24498078, year = {2014}, author = {Atyame, CM and Labbé, P and Dumas, E and Milesi, P and Charlat, S and Fort, P and Weill, M}, title = {Wolbachia divergence and the evolution of cytoplasmic incompatibility in Culex pipiens.}, journal = {PloS one}, volume = {9}, number = {1}, pages = {e87336}, pmid = {24498078}, issn = {1932-6203}, mesh = {Animals ; Crosses, Genetic ; Culex/classification/*genetics/microbiology ; Cytoplasm/genetics/microbiology ; Evolution, Molecular ; Female ; *Genetic Variation ; Host-Pathogen Interactions/genetics ; Male ; Models, Genetic ; Reproduction/genetics ; Species Specificity ; Wolbachia/classification/*genetics/physiology ; }, abstract = {Many insect species harbor Wolbachia bacteria that induce cytoplasmic incompatibility (CI), i.e. embryonic lethality in crosses between infected males and uninfected females, or between males and females carrying incompatible Wolbachia strains. The molecular mechanism of CI remains unknown, but the available data are best interpreted under a modification-rescue model, where a mod function disables the reproductive success of infected males' sperm, unless the eggs are infected and express a compatible resc function. Here we examine the evolution of CI in the mosquito Culex pipiens, harbouring a large number of closely related Wolbachia strains structured in five distinct phylogenetic groups. Specifically, we used a worldwide sample of mosquito lines to assess the hypothesis that genetic divergence should correlate with the divergence of CI properties on a low evolutionary scale. We observed a significant association of Wolbachia genetic divergence with CI patterns. Most Wolbachia strains from the same group were compatible whereas those from different groups were often incompatible. Consistently, we found a strong association between Wolbachia groups and their mod-resc properties. Finally, lines from the same geographical area were rarely incompatible, confirming the conjecture that the spatial distribution of Wolbachia compatibility types should be constrained by selection. This study indicates a clear correlation between Wolbachia genotypes and CI properties, paving the way toward the identification of the molecular basis of CI through comparative genomics.}, } @article {pmid24495395, year = {2014}, author = {Yeap, HL and Axford, JK and Popovici, J and Endersby, NM and Iturbe-Ormaetxe, I and Ritchie, SA and Hoffmann, AA}, title = {Assessing quality of life-shortening Wolbachia-infected Aedes aegypti mosquitoes in the field based on capture rates and morphometric assessments.}, journal = {Parasites & vectors}, volume = {7}, number = {}, pages = {58}, pmid = {24495395}, issn = {1756-3305}, mesh = {Aedes/anatomy & histology/microbiology/*physiology ; Animals ; Body Size ; Dengue/*prevention & control/virology ; Insect Vectors/anatomy & histology/microbiology/*physiology ; Mosquito Control/*methods ; Population Dynamics ; Reproduction ; Wings, Animal/anatomy & histology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Recent releases have been carried out with Aedes aegypti mosquitoes infected with the wMelPop mosquito cell-line adapted (wMelPop-CLA) strain of Wolbachia. This infection introduced from Drosophila provides strong blockage of dengue and other arboviruses but also has large fitness costs in laboratory tests. The releases were used to evaluate the fitness of released infected mosquitoes, and (following termination of releases) to test for any effects of wMelPop-CLA on wing size and shape when mosquitoes were reared under field conditions.

METHODS: We monitored gravid females via double sticky traps to assess the reproductive success of wMelPop-CLA-infected females and also sampled the overall mosquito population post-release using Biogent Sentinel traps. Morphometric analyses were used to evaluate infection effects on wing shape as well as size.

RESULTS: Oviposition success as assessed through double sticky traps was unrelated to size of released mosquitoes. However, released mosquitoes with lower wing loading were more successful. Furthermore, wMelPop-CLA-infected mosquitoes had 38.3% of the oviposition success of uninfected mosquitoes based on the predicted infection frequency after release. Environmental conditions affected wing shape and particularly size across time in uninfected mosquitoes, but not in naturally-reared wMelPop-CLA-infected mosquitoes. Although the overall size and shape do not differ between naturally-reared wMelPop-CLA-infected and uninfected mosquitoes, the infected mosquitoes tended to have smaller wings than uninfected mosquitoes during the cooler November in comparison to December.

CONCLUSION: These results confirm the lower fitness of wMelPop-CLA infection under field conditions, helping to explain challenges associated with a successful invasion by this strain. In the long run, invasion may depend on releasing strains carrying insecticide resistance or egg desiccation resistance, combined with an active pre-release population suppression program.}, } @article {pmid24488176, year = {2014}, author = {Murdock, CC and Blanford, S and Hughes, GL and Rasgon, JL and Thomas, MB}, title = {Temperature alters Plasmodium blocking by Wolbachia.}, journal = {Scientific reports}, volume = {4}, number = {}, pages = {3932}, pmid = {24488176}, issn = {2045-2322}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21 AI088311/AI/NIAID NIH HHS/United States ; R21 AI096036/AI/NIAID NIH HHS/United States ; R21 AI096036-01/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology/*parasitology ; Host-Parasite Interactions ; Insect Vectors/microbiology/parasitology ; Malaria/transmission ; Plasmodium yoelii/*growth & development/*microbiology ; Sporozoites/growth & development/microbiology ; Temperature ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Very recently, the Asian malaria vector (Anopheles stephensi) was stably transinfected with the wAlbB strain of Wolbachia, inducing refractoriness to the human malaria parasite Plasmodium falciparum. However, conditions in the field can differ substantially from those in the laboratory. We use the rodent malaria P. yoelii, and somatically transinfected An. stephensi as a model system to investigate whether the transmission blocking potential of wAlbB is likely to be robust across different thermal environments. wAlbB reduced malaria parasite prevalence and oocyst intensity at 28°C. At 24°C there was no effect on prevalence but a marked increase in oocyst intensity. At 20°C, wAlbB had no effect on prevalence or intensity. Additionally, we identified a novel effect of wAlbB that resulted in reduced sporozoite development across temperatures, counterbalancing the oocyst enhancement at 24°C. Our results demonstrate complex effects of temperature on the Wolbachia-malaria interaction, and suggest the impacts of transinfection might vary across diverse environments.}, } @article {pmid24484966, year = {2014}, author = {Bordbar, A and Soleimani, S and Fardid, F and Zolfaghari, MR and Parvizi, P}, title = {Three strains of Wolbachia pipientis and high rates of infection in Iranian sandfly species.}, journal = {Bulletin of entomological research}, volume = {104}, number = {2}, pages = {195-202}, doi = {10.1017/S0007485313000631}, pmid = {24484966}, issn = {1475-2670}, mesh = {Animals ; Female ; Iran ; Male ; Phlebotomus/*microbiology ; Wolbachia/*isolation & purification ; }, abstract = {Individual wild-caught sandflies from Iran were examined for infections of Wolbachia pipientis by targeting the major surface protein gene wsp of this intracellular α-proteobacterium. In total, 638 male and female sandflies were screened, of which 241 were found to be positive for one of three wsp haplotypes. Regardless of geographical origins and habitats, Phlebotomus (Phlebotomus) papatasi and other sandfly species were found to be infected with one common, widespread strain of A-group W. pipientis (Turk 54, GenBank accession EU780683; AY288297). In addition, a new A-group haplotype (Turk07, GenBank accession KC576916) was isolated from Phlebotomus (Paraphlebotomus) mongolensis and Phlebotomus (Pa.) caucasicus, and a new B-group haplotype (AZ2331, GenBank accession JX488735) was isolated from Phlebotomus (Larroussius) perfiliewi. Therefore, Wolbachia was found to occur in at least three of the incriminated vectors of zoonotic cutaneous leishmaniasis and zoonotic visceral leishmaniasis in different geographical regions of Iran. It may provide a new tool for the future control of leishmaniasis.}, } @article {pmid24481199, year = {2013}, author = {Bull, JJ and Turelli, M}, title = {Wolbachia versus dengue: Evolutionary forecasts.}, journal = {Evolution, medicine, and public health}, volume = {2013}, number = {1}, pages = {197-207}, pmid = {24481199}, issn = {2050-6201}, support = {R01 GM057756/GM/NIGMS NIH HHS/United States ; }, abstract = {A novel form of biological control is being applied to the dengue virus. The agent is the maternally transmitted bacterium Wolbachia, naturally absent from the main dengue vector, the mosquito Aedes aegypti. Three Wolbachia-based control strategies have been proposed. One is suppression of mosquito populations by large-scale releases of males incompatible with native females; this intervention requires ongoing releases. The other interventions transform wild mosquito populations with Wolbachia that spread via the frequency-dependent fitness advantage of Wolbachia-infected females; those interventions potentially require just a single, local release for area-wide disease control. One of these latter strategies uses Wolbachia that shortens mosquito life, indirectly preventing viral maturation/transmission. The other strategy uses Wolbachia that block viral transmission. All interventions can be undermined by viral, bacterial or mosquito evolution; viral virulence in humans may also evolve. We examine existing theory, experiments and comparative evidence to motivate predictions about evolutionary outcomes. (i) The life-shortening strategy seems the most likely to be thwarted by evolution. (ii) Mosquito suppression has a reasonable chance of working locally, at least in the short term, but long-term success over large areas is challenging. (iii) Dengue blocking faces strong selection for viral resistance but may well persist indefinitely at some level. Virulence evolution is not mathematically predictable, but comparative data provide no precedent for Wolbachia increasing dengue virulence. On balance, our analysis suggests that the considerable possible benefits of these technologies outweigh the known negatives, but the actual risk is largely unknown.}, } @article {pmid24480589, year = {2014}, author = {Arumugam, S and Hoerauf, A and Pfarr, KM}, title = {Localization of a filarial phosphate permease that is up-regulated in response to depletion of essential Wolbachia endobacteria.}, journal = {Experimental parasitology}, volume = {138}, number = {}, pages = {30-39}, doi = {10.1016/j.exppara.2014.01.006}, pmid = {24480589}, issn = {1090-2449}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Antibody Specificity ; Blotting, Western ; Doxycycline/pharmacology ; Female ; Filarioidea/*enzymology/genetics/microbiology ; Humans ; Immune Sera/immunology ; Immunohistochemistry ; Interleukin-5/deficiency ; Mice ; Mice, Inbred BALB C ; Onchocerca volvulus/drug effects/*enzymology/microbiology ; Phosphate Transport Proteins/immunology/isolation & purification/*metabolism ; Rabbits ; Tetracycline/pharmacology ; Up-Regulation ; Wolbachia/drug effects/*physiology ; }, abstract = {Wolbachia of filarial nematodes are essential, obligate endobacteria. When depleted by doxycycline worm embryogenesis, larval development and worm survival are inhibited. The molecular basis governing the endosymbiosis between Wolbachia and their filarial host is still being deciphered. In rodent filarial nematode Litomosoides sigmodontis, a nematode encoded phosphate permease gene (Ls-ppe-1) was up-regulated at the mRNA level in response to Wolbachia depletion and this gene promises to have an important role in Wolbachia-nematode endosymbiosis. To further characterize this gene, the regulation of phosphate permease during Wolbachia depletion was studied at the protein level in L. sigmodontis and in the human filaria Onchocerca volvulus. And the localization of phosphate permease (PPE) and Wolbachia in L. sigmodontis and O. volvulus was investigated in untreated and antibiotic treated worms. Depletion of Wolbachia by tetracycline (Tet) resulted in up-regulation of Ls-ppe-1 in L. sigmodontis. On day 36 of Tet treatment, compared to controls (Con), >98% of Wolbachia were depleted with a 3-fold increase in mRNA levels of Ls-ppe-1. Anti-Ls-PPE serum used in Western blots showed up-regulation of Ls-PPE at the protein level in Tet worms on day 15 and 36 of treatment. Immunohistology revealed the localization of Wolbachia and Ls-PPE in the embryos, microfilariae and hypodermis of L. sigmodontis female worms and up-regulation of Ls-PPE in response to Wolbachia depletion. Expression of O. volvulus phosphate permease (Ov-PPE) studied using anti-Ov-PPE serum, showed up-regulation of Ov-PPE at the protein level in doxycycline treated Wolbachia depleted O. volvulus worms and immunohistology revealed localization of Ov-PPE and Wolbachia and up-regulation of Ov-PPE in the hypodermis and embryos of doxycycline treated worms. Ls-PPE and Ov-PPE are upregulated upon Wolbachia depletion in same tissues and regions where Wolbachia are located in untreated worms, reinforcing a link between Wolbachia and this nematode encoded protein. The function of nematode phosphate permease in the endosymbiosis is unknown but could involve transportation of phosphate to Wolbachia, which encode all the genes necessary for de novo nucleotide biosynthesis. Electron microscopic localization of PPE and Wolbachia and RNAi mediated knock-down of PPE in filarial nematodes will bring further insights to the functions of PPE in the Wolbachia-nematode symbiosis.}, } @article {pmid24479705, year = {2014}, author = {Ali, M and Afzal, M and Abdul Nasim, S and Ahmad, I}, title = {Nanocurcumin: a novel antifilarial agent with DNA topoisomerase II inhibitory activity.}, journal = {Journal of drug targeting}, volume = {22}, number = {5}, pages = {395-407}, doi = {10.3109/1061186X.2013.869823}, pmid = {24479705}, issn = {1029-2330}, mesh = {Animals ; Brugia/drug effects/enzymology/physiology ; Curcumin/administration & dosage/pharmacokinetics/*therapeutic use ; Disease Models, Animal ; Drug Carriers/*chemistry ; Drug Liberation ; Elephantiasis, Filarial/*drug therapy/parasitology ; Filaricides/administration & dosage/*therapeutic use ; Host-Parasite Interactions/physiology ; Male ; Mice ; Nanoparticles/*chemistry ; Particle Size ; Rats ; Surface Properties ; Tissue Distribution ; Topoisomerase II Inhibitors/administration & dosage/pharmacokinetics/*therapeutic use ; }, abstract = {OBJECTIVE: The aim of this study is to evaluate the antifilarial, antiwolbachial and DNA topoisomerase II inhibitory activity of nanocurcumin (nano-CUR).

METHODS: Nano-CUR formulations (F1-F6) were prepared using free radical polymerization and were characterized by particle size, morphology, encapsulation efficiency and in vitro release kinetics. Antifilarial potential was evaluated in vivo against Brugian filariasis in an experimental rodent model, Mastomys coucha, by selecting the formulation that maximized parasite elimination characteristics. Wolbachial status was determined by PCR and a relaxation assay was used to estimate DNA topoisomerase II inhibitory activity.

RESULTS: Nano-CUR (F3) having a 60 nm diameter and 89.78% entrapment efficiency showed the most favorable characteristics for the elimination of filarial parasites. In vivo pharmacokinetic and organ distribution studies demonstrate significantly greater C(max) (86.6 ± 2.56 ng ml(-1)), AUC0-∞ (796 ± 89.8 ng d ml(-1)), MRT (19.5 ± 7.82 days) and bioavailability of CUR (70.02%) in the organs from which the adult parasites were recovered. The optimized nano-CUR (F3) (5 × 5 mg/kg, orally) significantly augmented the microfilariciadal and adulticidal action of CUR over free CUR (5 × 50 mg/kg, orally) or Diethylcarbamizine (50 mg/kg, orally) against the Brugia malayi Mastomys coucha rodent model. The PCR results showed complete elimination of wolbachia from the recovered female parasites. Interestingly, nano-CUR was also found to be a novel inhibitor of filarial worm DNA topoisomerase II, Setaria Cervi in vitro.

CONCLUSION: This study recognizes the beforehand antimicrofilarial, antimacrofilarial, anti-wolbachial activity of nano-CUR (F3) over free forms and additionally its strong inhibitory action against the major target filarial parasite enzyme DNA topoisomerase II in vitro.}, } @article {pmid24472200, year = {2014}, author = {De Clerck, C and Tsuchida, T and Massart, S and Lepoivre, P and Francis, F and Jijakli, MH}, title = {Combination of genomic and proteomic approaches to characterize the symbiotic population of the banana aphid (Hemiptera: Aphididae).}, journal = {Environmental entomology}, volume = {43}, number = {1}, pages = {29-36}, doi = {10.1603/EN13107}, pmid = {24472200}, issn = {1938-2936}, mesh = {Animals ; Aphids/genetics/*microbiology ; Babuvirus/genetics ; Buchnera/isolation & purification ; DNA, Bacterial/*chemistry/genetics ; Genomics ; Musa ; Symbiosis/genetics ; Wolbachia/isolation & purification ; }, abstract = {Aphids are known to live in symbiosis with specific bacteria called endosymbionts that have positive or negative impacts on their hosts. In this study, six banana aphid (Pentalonia nigronervosa Coquerel) strains from various geographical origins (Gabon, Madagascar, and Burundi) were screened to determine their symbiotic content, using complementary genomic (16S rDNA sequencing and specific polymerase chain reaction) and proteomic (two-dimensional difference gel electrophoresis coupled with protein identification by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry) approaches. Despite the geographical heterogeneity, the combined methods allowed us to identify the same two symbionts in the six aphids strains tested: Buchnera aphidicola and Wolbachia. Although B. aphidicola is found in almost all aphid species, the systematic presence of Wolbachia in banana aphids is particularly interesting, as this bacterium usually has a low prevalence in aphid species. Phylogenetic analyses showed that the Wolbachia sp. strain found in P. nigronervosa was very similar to the strain present in aphids of the genus Cinara, known to have developed a strong and long-term symbiotic association with Wolbachia. The high level of asexual reproduction in P. nigronervosa could be linked to the presence of Wolbachia, but its prevalence also suggests that this symbiotic bacterium could play a more essential role in its aphid host.}, } @article {pmid24472097, year = {2014}, author = {Dupeyron, M and Leclercq, S and Cerveau, N and Bouchon, D and Gilbert, C}, title = {Horizontal transfer of transposons between and within crustaceans and insects.}, journal = {Mobile DNA}, volume = {5}, number = {1}, pages = {4}, pmid = {24472097}, issn = {1759-8753}, abstract = {BACKGROUND: Horizontal transfer of transposable elements (HTT) is increasingly appreciated as an important source of genome and species evolution in eukaryotes. However, our understanding of HTT dynamics is still poor in eukaryotes because the diversity of species for which whole genome sequences are available is biased and does not reflect the global eukaryote diversity.

RESULTS: In this study we characterized two Mariner transposable elements (TEs) in the genome of several terrestrial crustacean isopods, a group of animals particularly underrepresented in genome databases. The two elements have a patchy distribution in the arthropod tree and they are highly similar (>93% over the entire length of the element) to insect TEs (Diptera and Hymenoptera), some of which were previously described in Ceratitis rosa (Crmar2) and Drosophila biarmipes (Mariner-5_Dbi). In addition, phylogenetic analyses and comparisons of TE versus orthologous gene distances at various phylogenetic levels revealed that the taxonomic distribution of the two elements is incompatible with vertical inheritance.

CONCLUSIONS: We conclude that the two Mariner TEs each underwent at least three HTT events. Both elements were transferred once between isopod crustaceans and insects and at least once between isopod crustacean species. Crmar2 was also transferred between tephritid and drosophilid flies and Mariner-5 underwent HT between hymenopterans and dipterans. We demonstrate that these various HTTs took place recently (most likely within the last 3 million years), and propose iridoviruses and/or Wolbachia endosymbionts as potential vectors of these transfers.}, } @article {pmid24468556, year = {2013}, author = {Pandey, N and Singh, A and Rana, VS and Rajagopal, R}, title = {Molecular characterization and analysis of bacterial diversity in Aleurocanthus woglumi (Hemiptera: Aleyrodidae).}, journal = {Environmental entomology}, volume = {42}, number = {6}, pages = {1257-1264}, doi = {10.1603/EN13110}, pmid = {24468556}, issn = {1938-2936}, mesh = {Animals ; Bacteria/chemistry/genetics ; *Biodiversity ; DNA, Bacterial/chemistry ; Electron Transport Complex IV/genetics ; Hemiptera/enzymology/genetics/*microbiology ; *Microbial Consortia ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; }, abstract = {Aleurocanthus woglumi Ashby (Hemiptera: Aleyrodidae), commonly referred to as citrus blackfly, is a sap-sucking hemipteran insect. Although polyphagous, citrus is its most preferred host plant. Samples of this insect were collected from Murraya koenigii (L.). The cytochrome c oxidase subunit I gene (mtCO1)-based analysis by sequencing helped in molecular identification of the insect. Phylogenetic analysis of cytB-nd1-LrDNA showed the coevolution of A. woglumi with its primary bacterial symbiont Portiera. Sequencing a 16S rDNA library from insect DNA revealed three bacterial phylotypes, namely, Portiera, Wolbachia, and Erwinia chrysanthemi. Further, we used fluorescence in situ hybridization to visualize the endosymbionts in a whole mount of A. woglumi. Culturable bacteria were obtained on different media and were classified on the basis of 16S rDNA. In total, 30 bacterial phylotypes belonging to 14 different genera, namely, Bacillus, Kocuria, Micrococcus, Staphylococcus, Paenibacillus, Rhodococcus, Rummellibacillus, Arthrobacter, Curtobacterium, Psychrobacillus, Listeria, Brevibacillus, Bhargavae, and Pantoea, were isolated by culturable methods.}, } @article {pmid24466066, year = {2014}, author = {Fischer, K and Beatty, WL and Weil, GJ and Fischer, PU}, title = {High pressure freezing/freeze substitution fixation improves the ultrastructural assessment of Wolbachia endosymbiont-filarial nematode host interaction.}, journal = {PloS one}, volume = {9}, number = {1}, pages = {e86383}, pmid = {24466066}, issn = {1932-6203}, support = {S10 RR031552/RR/NCRR NIH HHS/United States ; S10RR031552/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/growth & development/*microbiology/*ultrastructure ; Female ; Freeze Substitution/*methods ; *Freezing ; *Host-Pathogen Interactions ; Life Cycle Stages ; Parasites/microbiology/ultrastructure ; Pressure ; Secretory Vesicles/metabolism/ultrastructure ; *Symbiosis ; Wolbachia/*ultrastructure ; }, abstract = {BACKGROUND: Wolbachia α-proteobacteria are essential for growth, reproduction and survival for many filarial nematode parasites of medical and veterinary importance. Endobacteria were discovered in filarial parasites by transmission electron microscopy in the 1970's using chemically fixed specimens. Despite improvements of fixation and electron microscopy techniques during the last decades, methods to study the Wolbachia/filaria interaction on the ultrastructural level remained unchanged and the mechanisms for exchange of materials and for motility of endobacteria are not known.

METHODOLOGY/PRINCIPAL FINDING: We used high pressure freezing/freeze substitution to improve fixation of Brugia malayi and its endosymbiont, and this led to improved visualization of different morphological forms of Wolbachia. The three concentric, bilayer membranes that surround the endobacterial cytoplasm were well preserved. Vesicles with identical membrane structures were identified close to the endobacteria, and multiple bacteria were sometimes enclosed within a single outer membrane. Immunogold electron microscopy using a monoclonal antibody directed against Wolbachia surface protein-1 labeled the membranes that enclose Wolbachia and Wolbachia-associated vesicles. High densities of Wolbachia were observed in the lateral chords of L4 larvae, immature, and mature adult worms. Extracellular Wolbachia were sometimes present in the pseudocoelomic cavity near the developing female reproductive organs. Wolbachia-associated actin tails were not observed. Wolbachia motility may be explained by their residence within vacuoles, as they may co-opt the host cell's secretory pathway to move within and between cells.

CONCLUSIONS/SIGNIFICANCE: High pressure freezing/freeze substitution significantly improved the preservation of filarial tissues for electron microscopy to reveal membranes and sub cellular structures that could be crucial for exchange of materials between Wolbachia and its host.}, } @article {pmid24465416, year = {2014}, author = {Guo, H and Qu, Y and Liu, X and Zhong, W and Fang, J}, title = {Female-biased symbionts and tomato yellow leaf curl virus infections in Bemisia tabaci.}, journal = {PloS one}, volume = {9}, number = {1}, pages = {e84538}, pmid = {24465416}, issn = {1932-6203}, mesh = {Animals ; Bacteria/classification/*genetics/growth & development ; Begomovirus/*genetics/physiology ; DNA, Bacterial/genetics ; DNA, Viral/genetics ; Enterobacteriaceae/genetics/physiology ; Female ; Hemiptera/*genetics/microbiology/virology ; Host-Pathogen Interactions ; Solanum lycopersicum/virology ; Male ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 23S/genetics ; Rickettsia/genetics/physiology ; Sex Factors ; Species Specificity ; *Symbiosis ; Wolbachia/genetics/physiology ; }, abstract = {The female-biased infection of facultative symbionts has been found in Bemisia tabaci; however, whether there are any differences in tomato yellow leaf curl virus (TYLCV) and obligate symbiont infection rates between females and males is unknown. Determining whether such differences exist would be very important for understanding the spread of the plant virus and of the symbionts. We compared both symbiont infection types, including obligate and facultative symbionts, and the rates of TYLCV infection in both sexes in five field populations from Jiangsu Province, China. The obligate symbiont Portiera aleyrodidarum was not found in every whitefly tested. In all tested populations, more females than males were found to harbor P. aleyrodidarum; and more females than males also harbored Hamiltonella defense, the most common facultative symbiont as well as Cardinium. In addition to female-biased symbiont infections, there were also female-biased TYLCV infections, and the infection frequencies of this plant virus in females were higher than those in males. Taken together, these results suggested that both the female-biased symbiont infections and female-biased TYLCV infections promoted the rapid spread of TYLCV in China.}, } @article {pmid24463012, year = {2014}, author = {Cass, BN and Mozes-Daube, N and Iasur-Kruh, L and Bondy, EC and Kelly, SE and Hunter, MS and Zchori-Fein, E}, title = {Bacterial endosymbionts in field-collected samples of Trialeurodes sp. nr. abutiloneus (Hemiptera: Aleyrodidae).}, journal = {Research in microbiology}, volume = {165}, number = {2}, pages = {77-81}, doi = {10.1016/j.resmic.2014.01.005}, pmid = {24463012}, issn = {1769-7123}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; *Bacterial Physiological Phenomena ; Gossypium/parasitology ; Hemiptera/*microbiology ; Molecular Sequence Data ; Sequence Analysis, DNA ; *Symbiosis ; United States ; }, abstract = {Facultative bacterial endosymbionts are common, influential associates of arthropods, yet their movement among host species has not been well documented. Plant-mediated transmission of Rickettsia has been shown for the whitefly Bemisia tabaci. Bemisia tabaci in USA cotton fields harbors the secondary symbionts Rickettsia and Hamiltonella, and co-occurs with Trialeurodes sp. nr. abutiloneus whiteflies. To determine whether symbionts may be shared, the microbial diversity of these whiteflies on cotton across the USA was analyzed. Trialeurodes sp. nr. abutiloneus bore Portiera, Pseudomonas, Serratia, Arsenophonus and Wolbachia. No Rickettsia or Hamiltonella were detected. These results provide no evidence for horizontal transmission of symbionts between these whitefly genera.}, } @article {pmid24461455, year = {2014}, author = {Le, PT and Pontarotti, P and Raoult, D}, title = {Alphaproteobacteria species as a source and target of lateral sequence transfers.}, journal = {Trends in microbiology}, volume = {22}, number = {3}, pages = {147-156}, doi = {10.1016/j.tim.2013.12.006}, pmid = {24461455}, issn = {1878-4380}, mesh = {Alphaproteobacteria/*genetics ; Bacterial Secretion Systems ; Biological Evolution ; *Gene Transfer, Horizontal ; }, abstract = {Alphaproteobacterial genomes show a remarkable genome plasticity linked with different lifestyles (intracellular, facultative, and free-living). They represent the major source of the genome repertoire of mitochondria, and their genes (specifically those of Wolbachia) have been massively transferred into their modern eukaryotic hosts, such as arthropods and nematodes. Conversely, other organisms (bacteria, viruses, archaea, and eukaryotes) and selfish DNA have contributed to their genomes. This bidirectional lateral sequence transfer explains the mosaic nature of their genomes. In contrast to those living in allopatry, alphaproteobacteria living in sympatry (in protist cells such as in the environment) favor lateral sequence transfer. Evidence shows that intracellular transfer of the type IV secretion system might have played a critical role in the evolution of these alphaproteobacteria.}, } @article {pmid24458589, year = {2014}, author = {Gauthier, N and Clouet, C and Perrakis, A and Kapantaidaki, D and Peterschmitt, M and Tsagkarakou, A}, title = {Genetic structure of Bemisia tabaci Med populations from home-range countries, inferred by nuclear and cytoplasmic markers: impact on the distribution of the insecticide resistance genes.}, journal = {Pest management science}, volume = {70}, number = {10}, pages = {1477-1491}, doi = {10.1002/ps.3733}, pmid = {24458589}, issn = {1526-4998}, mesh = {Animals ; Base Sequence ; Female ; Gene Flow ; Genetics, Population ; Hemiptera/*classification/*genetics/microbiology ; Insecticide Resistance/*genetics ; Insecticides/*pharmacology ; Mediterranean Region ; Microsatellite Repeats ; Organophosphates/*pharmacology ; Phylogeography ; Pyrethrins/*pharmacology ; Symbiosis ; Wolbachia ; }, abstract = {BACKGROUND: Insecticide resistance management in Bemisia tabaci is one of the main issues facing agricultural production today. An extensive survey was undertaken in five Mediterranean countries to examine the resistance status of Med B. tabaci species in its range of geographic origin and the relationship between population genetic structure and the distribution of resistance genes. The investigation combined molecular diagnostic tests, sequence and microsatellite polymorphism studies and monitoring of endosymbionts.

RESULTS: High frequencies of pyrethroid (L925I and T929V, VGSC gene) and organophosphate (F331W, ace1 gene) resistance mutations were found in France, Spain and Greece, but not in Morocco or Tunisia. Sequence analyses of the COI gene delineated two closely related mitochondrial groups (Q1 and Q2), which were found either sympatrically (Spain) or separately (France). Only Q1 was observed in Greece, Morocco and Tunisia. Bayesian analyses based on microsatellite loci revealed three geographically delineated genetic groups (France, Spain, Morocco/Greece/Tunisia) and high levels of genetic differentiation even between neighbouring samples. Evidence was also found for hybridisation and asymmetrical gene flow between Q1 and Q2.

CONCLUSIONS: Med B. tabaci is more diverse and structured than reported so far. On a large geographic scale, resistance is affected by population genetic structure, whereas on a local scale, agricultural practices appear to play a major role.}, } @article {pmid24455146, year = {2013}, author = {Kodandaramaiah, U and Simonsen, TJ and Bromilow, S and Wahlberg, N and Sperling, F}, title = {Deceptive single-locus taxonomy and phylogeography: Wolbachia-associated divergence in mitochondrial DNA is not reflected in morphology and nuclear markers in a butterfly species.}, journal = {Ecology and evolution}, volume = {3}, number = {16}, pages = {5167-5176}, pmid = {24455146}, issn = {2045-7758}, abstract = {The satyrine butterfly Coenonympha tullia (Nymphalidae: Satyrinae) displays a deep split between two mitochondrial clades, one restricted to northern Alberta, Canada, and the other found throughout Alberta and across North America. We confirm this deep divide and test hypotheses explaining its phylogeographic structure. Neither genitalia morphology nor nuclear gene sequence supports cryptic species as an explanation, instead indicating differences between nuclear and mitochondrial genome histories. Sex-biased dispersal is unlikely to cause such mito-nuclear differences; however, selective sweeps by reproductive parasites could have led to this conflict. About half of the tested samples were infected by Wolbachia bacteria. Using multilocus strain typing for three Wolbachia genes, we show that the divergent mitochondrial clades are associated with two different Wolbachia strains, supporting the hypothesis that the mito-nuclear differences resulted from selection on the mitochondrial genome due to selective sweeps by Wolbachia strains.}, } @article {pmid24428638, year = {2014}, author = {Morrow, JL and Frommer, M and Shearman, DC and Riegler, M}, title = {Tropical tephritid fruit fly community with high incidence of shared Wolbachia strains as platform for horizontal transmission of endosymbionts.}, journal = {Environmental microbiology}, volume = {16}, number = {12}, pages = {3622-3637}, doi = {10.1111/1462-2920.12382}, pmid = {24428638}, issn = {1462-2920}, mesh = {Animals ; Australia ; Female ; Incidence ; Male ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny ; Polymerase Chain Reaction ; *Symbiosis ; Tephritidae/genetics/*microbiology/physiology ; Wasps/microbiology ; Wolbachia/*classification/genetics/*isolation & purification/physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria that infect 40-65% of arthropod species. They are primarily maternally inherited with occasional horizontal transmission for which limited direct ecological evidence exists. We detected Wolbachia in 8 out of 24 Australian tephritid species. Here, we have used multilocus sequence typing (MLST) to further characterize these Wolbachia strains, plus a novel quantitative polymerase chain reaction method for allele assignment in multiple infections. Based on five MLST loci and the Wolbachia surface protein gene (wsp), five Bactrocera and one Dacus species harboured two identical strains as double infections; furthermore, Bactrocera neohumeralis harboured both of these as single or double infections, and sibling species B. tryoni harboured one. Two Bactrocera species contained Wolbachia pseudogenes, potentially within the fruit fly genomes. A fruit fly parasitoid, Fopius arisanus shared identical alleles with two Wolbachia strains detected in one B. frauenfeldi individual. We report an unprecedented high incidence of four shared Wolbachia strains in eight host species from two trophic levels. This suggests frequent exposure to Wolbachia in this tropical tephritid community that shares host plant and parasitoid species, and also includes species that hybridize. Such insect communities may act as horizontal transmission platforms that contribute to the ubiquity of the otherwise maternally inherited Wolbachia.}, } @article {pmid24402369, year = {2014}, author = {Perlman, SJ and Dowdy, NJ and Harris, LR and Khalid, M and Kelly, SE and Hunter, MS}, title = {Factors affecting the strength of Cardinium-induced cytoplasmic incompatibility in the parasitic wasp Encarsia pergandiella (Hymenoptera: Aphelinidae).}, journal = {Microbial ecology}, volume = {67}, number = {3}, pages = {671-678}, pmid = {24402369}, issn = {1432-184X}, mesh = {Animals ; Bacteroidetes/genetics/*physiology ; Cytoplasm/physiology ; Female ; Male ; Reproduction ; *Symbiosis ; Wasps/genetics/growth & development/*microbiology/*physiology ; }, abstract = {Bacteria that cause cytoplasmic incompatibility (CI) are among the most common maternally transmitted parasites of insects. In CI, uninfected females produce few or no offspring when they mate with infected males and, as a result, are often at a reproductive disadvantage relative to infected females. Two different bacteria are known to cause CI, Wolbachia and Cardinium. CI Cardinium was discovered more recently and has been little studied. Here, factors that could influence the reduction in reproductive output in a CI cross, or CI "strength," were explored in the parasitic wasp Encarsia pergandiella. Cardinium in this wasp exhibits variable CI strength. Experiments tested the effect of male age, male size, male host species, Cardinium density, and male development time on CI strength. We found a striking effect of male development time, with males that took longer to develop exhibiting stronger CI when mated to uninfected females. Male age had little effect; although in one experiment, the oldest males exhibited stronger CI. Male size, host species, and bacterial density had no effect on the strength of CI. Identifying the factors that control CI are crucial for understanding the dynamics of infection, as well as the success of strategies that aim to use CI microbes to control insect pests and disease vectors.}, } @article {pmid24395812, year = {2014}, author = {Wang, Z and Su, XM and Wen, J and Jiang, LY and Qiao, GX}, title = {Widespread infection and diverse infection patterns of Wolbachia in Chinese aphids.}, journal = {Insect science}, volume = {21}, number = {3}, pages = {313-325}, doi = {10.1111/1744-7917.12102}, pmid = {24395812}, issn = {1744-7917}, mesh = {Animals ; Aphids/*microbiology ; Genetic Variation ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are intracellular symbionts that infect a wide range of arthropods and filarial nematodes. Aphids are engaged in diverse and complex relationships with their endosymbionts. Four supergroups (A, B, M and N) of Wolbachia were previously detected in aphids and supergroups M and N were only found in aphids. In this study, we detected and described Wolbachia infections in natural populations of aphids in China. Three supergroups (A, B and M) were found in the examined aphid species. Supergroup M was preponderant, whereas supergroups A and B were only detected in certain species. Supergroup N was not found in this study. There were four infection patterns of Wolbachia in aphids, namely, infection with supergroup M alone, co-infection with supergroups A and M, co-infection with supergroups B and M, and co-infection with supergroups A, B and M. The pattern of infection only with supergroup M was universal and was found in all evaluated subfamilies. Only two subfamilies, Aphidinae and Lachninae, manifested to present all four infection patterns. Three patterns were observed in Calaphidinae (M, A&M, B&M) and Eriosomatinae (M, B&M, A&B&M). Two patterns were observed in the Anoeciinae (M, A&M) and Greenideinae (M, B&M), and only one pattern (M) was observed in the remaining families and/or subfamilies of Aphidoidea. These results indicated that Wolbachia infections in Chinese aphids are widespread. Phylogenetic analyses suggest that Wolbachia supergroup M spread rapidly and recently among all host species of aphids in China. Reasons for this spread and its mechanisms are discussed along with the possible effects of Wolbachia on their aphid hosts.}, } @article {pmid24387805, year = {2014}, author = {Versace, E and Nolte, V and Pandey, RV and Tobler, R and Schlötterer, C}, title = {Experimental evolution reveals habitat-specific fitness dynamics among Wolbachia clades in Drosophila melanogaster.}, journal = {Molecular ecology}, volume = {23}, number = {4}, pages = {802-814}, pmid = {24387805}, issn = {1365-294X}, support = {W 1225/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; DNA, Mitochondrial/genetics ; Drosophila melanogaster/genetics/*microbiology ; *Ecosystem ; *Evolution, Molecular ; Female ; *Genetic Fitness ; Phylogeny ; Polymorphism, Single Nucleotide ; Temperature ; Wolbachia/*genetics ; }, abstract = {The diversity and infection dynamics of the endosymbiont Wolbachia can be influenced by many factors, such as transmission rate, cytoplasmic incompatibility, environment, selection and genetic drift. The interplay of these factors in natural populations can result in heterogeneous infection patterns with substantial differences between populations and strains. The causes of these heterogeneities are not yet understood, partly due to the complexity of natural environments. We present experimental evolution as a new approach to study Wolbachia infection dynamics in replicate populations exposed to a controlled environment. A natural Drosophila melanogaster population infected with strains of Wolbachia belonging to different clades evolved in two laboratory environments (hot and cold) for 1.5 years. In both treatments, the rate of Wolbachia infection increased until fixation. In the hot environment, the relative frequency of different Wolbachia clades remained stable over 37 generations. In the cold environment, however, we observed marked changes in the composition of the Wolbachia population: within 15 generations, one Wolbachia clade increased more than 50% in frequency, whereas the other two clades decreased in frequency, resulting in the loss of one clade. The frequency change was highly reproducible not only among replicates, but also when flies that evolved for 42 generations in the hot environment were transferred to the cold environment. These results document how environmental factors can affect the composition of Wolbachia in D. melanogaster. The high reproducibility of the pattern suggests that experimental evolution studies can efficiently determine the functional basis of habitat-specific fitness among Wolbachia strains.}, } @article {pmid24385937, year = {2013}, author = {Miller, WJ}, title = {Bugs in transition: the dynamic world of Wolbachia in insects.}, journal = {PLoS genetics}, volume = {9}, number = {12}, pages = {e1004069}, pmid = {24385937}, issn = {1553-7404}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology ; Symbiosis/*genetics ; Wolbachia/*genetics/growth & development ; }, } @article {pmid24385901, year = {2013}, author = {Hamilton, PT and Perlman, SJ}, title = {Host defense via symbiosis in Drosophila.}, journal = {PLoS pathogens}, volume = {9}, number = {12}, pages = {e1003808}, pmid = {24385901}, issn = {1553-7374}, mesh = {Aedes/microbiology ; Animals ; Disease Vectors ; Drosophila melanogaster/*immunology/*microbiology ; Female ; Male ; Spiroplasma/physiology ; Symbiosis/*physiology ; Wolbachia/physiology ; }, } @article {pmid24383453, year = {2013}, author = {Greiman, SE and Tkach, VV and Vaughan, JA}, title = {Transmission rates of the bacterial endosymbiont, Neorickettsia risticii, during the asexual reproduction phase of its digenean host, Plagiorchis elegans, within naturally infected lymnaeid snails.}, journal = {Parasites & vectors}, volume = {6}, number = {}, pages = {303}, pmid = {24383453}, issn = {1756-3305}, support = {R15AI092622/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Host-Pathogen Interactions ; Neorickettsia risticii/*physiology ; Reproduction ; Snails/*parasitology ; Trematoda/*microbiology ; }, abstract = {BACKGROUND: Neorickettsia are obligate intracellular bacterial endosymbionts of digenean parasites present in all lifestages of digeneans. Quantitative information on the transmission of neorickettsial endosymbionts throughout the complex life cycles of digeneans is lacking. This study quantified the transmission of Neorickettsia during the asexual reproductive phase of a digenean parasite, Plagiorchis elegans, developing within naturally parasitized lymnaeid pond snails.

METHODS: Lymnaea stagnalis snails were collected from 3 ponds in Nelson County, North Dakota and screened for the presence of digenean cercariae. Cercariae were identified to species by PCR and sequencing of the 28S rRNA gene. Neorickettsia infections were initially detected using nested PCR and sequencing of a partial 16S rRNA gene of pooled cercariae shed from each parasitized snail. Fifty to 100 single cercariae or sporocysts were isolated from each of six parasitized snails and tested for the presence of Neorickettsia using nested PCR to estimate the efficiency at which Neorickettsia were transmitted to cercariae during asexual development of the digenean.

RESULTS: A total of 616 L. stagnalis were collected and 240 (39%) shed digenean cercariae. Of these, 18 (8%) were Neorickettsia-positive. Six Neorickettsia infections were selected to determine the transmission efficiency of Neorickettsia from mother to daughter sporocyst and from daughter sporocyst to cercaria. The prevalence of neorickettsiae in cercariae varied from 11 to 91%. The prevalence of neorickettsiae in sporocysts from one snail was 100%.

CONCLUSION: Prevalence of Neorickettsia infection in cercariae of Plagiorchis elegans was variable and never reached 100%. Reasons for this are speculative, however, the low prevalence of Neorickettsia observed in some of our samples (11 to 52%) differs from the high prevalence of other, related bacterial endosymbionts, e.g. Wolbachia in Wolbachia-dependent filariid nematodes, where the prevalence among progeny is universally 100%. This suggests that, unlike the Wolbachia-filaria relationship, the Neorickettsia-digenean relationship is not obligatory mutualism. Our study represents the first quantitative estimate of the Neorickettsia transmission through the asexual phase of the digenean life cycle.}, } @article {pmid24376692, year = {2013}, author = {Maccari, M and Amat, F and Gómez, A}, title = {Origin and genetic diversity of diploid parthenogenetic Artemia in Eurasia.}, journal = {PloS one}, volume = {8}, number = {12}, pages = {e83348}, pmid = {24376692}, issn = {1932-6203}, mesh = {Animals ; Artemia/*classification/genetics ; Asia ; *Biological Evolution ; Cell Nucleus/genetics ; DNA, Intergenic/classification/genetics ; *Diploidy ; Europe ; Female ; Genes, Mitochondrial ; *Genetic Variation ; Male ; Mitochondria/genetics ; Parthenogenesis/*genetics ; *Phylogeny ; Phylogeography ; Sodium-Potassium-Exchanging ATPase/classification/genetics ; }, abstract = {There is wide interest in understanding how genetic diversity is generated and maintained in parthenogenetic lineages, as it will help clarify the debate of the evolution and maintenance of sexual reproduction. There are three mechanisms that can be responsible for the generation of genetic diversity of parthenogenetic lineages: contagious parthenogenesis, repeated hybridization and microorganism infections (e.g. Wolbachia). Brine shrimps of the genus Artemia (Crustacea, Branchiopoda, Anostraca) are a good model system to investigate evolutionary transitions between reproductive systems as they include sexual species and lineages of obligate parthenogenetic populations of different ploidy level, which often co-occur. Diploid parthenogenetic lineages produce occasional fully functional rare males, interspecific hybridization is known to occur, but the mechanisms of origin of asexual lineages are not completely understood. Here we sequenced and analysed fragments of one mitochondrial and two nuclear genes from an extensive set of populations of diploid parthenogenetic Artemia and sexual species from Central and East Asia to investigate the evolutionary origin of diploid parthenogenetic Artemia, and geographic origin of the parental taxa. Our results indicate that there are at least two, possibly three independent and recent maternal origins of parthenogenetic lineages, related to A. urmiana and Artemia sp. from Kazakhstan, but that the nuclear genes are very closely related in all the sexual species and parthenogegetic lineages except for A. sinica, who presumable took no part on the origin of diploid parthenogenetic strains. Our data cannot rule out either hybridization between any of the very closely related Asiatic sexual species or rare events of contagious parthenogenesis via rare males as the contributing mechanisms to the generation of genetic diversity in diploid parthenogenetic Artemia lineages.}, } @article {pmid24376534, year = {2013}, author = {Schneider, DI and Riegler, M and Arthofer, W and Merçot, H and Stauffer, C and Miller, WJ}, title = {Uncovering Wolbachia diversity upon artificial host transfer.}, journal = {PloS one}, volume = {8}, number = {12}, pages = {e82402}, pmid = {24376534}, issn = {1932-6203}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Amino Acids/genetics ; Animals ; Base Sequence ; Ceratitis capitata/*microbiology ; Codon, Terminator/genetics ; Conserved Sequence ; Drosophila/*microbiology ; Female ; Gene Frequency/genetics ; *Gene Transfer Techniques ; Genes, Bacterial ; *Genetic Variation ; *Host Specificity ; Molecular Sequence Data ; Nucleotides/genetics ; Ovary/microbiology ; Polymorphism, Single Nucleotide/genetics ; Synteny/genetics ; Wolbachia/*genetics ; }, abstract = {The common endosymbiotic Wolbachia bacteria influence arthropod hosts in multiple ways. They are mostly recognized for their manipulations of host reproduction, yet, more recent studies demonstrate that Wolbachia also impact host behavior, metabolic pathways and immunity. Besides their biological and evolutionary roles, Wolbachia are new potential biological control agents for pest and vector management. Importantly, Wolbachia-based control strategies require controlled symbiont transfer between host species and predictable outcomes of novel Wolbachia-host associations. Theoretically, this artificial horizontal transfer could inflict genetic changes within transferred Wolbachia populations. This could be facilitated through de novo mutations in the novel recipient host or changes of haplotype frequencies of polymorphic Wolbachia populations when transferred from donor to recipient hosts. Here we show that Wolbachia resident in the European cherry fruit fly, Rhagoletis cerasi, exhibit ancestral and cryptic sequence polymorphism in three symbiont genes, which are exposed upon microinjection into the new hosts Drosophila simulans and Ceratitis capitata. Our analyses of Wolbachia in microinjected D. simulans over 150 generations after microinjection uncovered infections with multiple Wolbachia strains in trans-infected lines that had previously been typed as single infections. This confirms the persistence of low-titer Wolbachia strains in microinjection experiments that had previously escaped standard detection techniques. Our study demonstrates that infections by multiple Wolbachia strains can shift in prevalence after artificial host transfer driven by either stochastic or selective processes. Trans-infection of Wolbachia can claim fitness costs in new hosts and we speculate that these costs may have driven the shifts of Wolbachia strains that we saw in our model system.}, } @article {pmid24373187, year = {2014}, author = {Bryson, RW}, title = {Bacterial endosymbiont infections in 'living fossils': a case study of North American vaejovid scorpions.}, journal = {Molecular ecology resources}, volume = {14}, number = {4}, pages = {789-793}, doi = {10.1111/1755-0998.12220}, pmid = {24373187}, issn = {1755-0998}, mesh = {Animals ; Bacteria/genetics/*isolation & purification ; *Bacterial Physiological Phenomena ; DNA Primers/genetics ; Genetic Testing/methods ; Scorpions/*microbiology ; *Symbiosis ; }, abstract = {Bacterial endosymbionts are common among arthropods, and maternally inherited forms can affect the reproductive and behavioural traits of their arthropod hosts. The prevalence of bacterial endosymbionts and their role in scorpion evolution have rarely been investigated. In this study, 61 samples from 40 species of scorpion in the family Vaejovidae were screened for the presence of the bacterial endosymbionts Cardinium, Rickettsia, Spiroplasma and Wolbachia. No samples were infected by these bacteria. However, one primer pair specifically designed to amplify Rickettsia amplified nontarget genes of other taxa. Similar off-target amplification using another endosymbiont-specific primer was also found during preliminary screenings. Results caution against the overreliance on previously published screening primers to detect bacterial endosymbionts in host taxa and suggest that primer specificity may be higher in primers targeting nuclear rather than mitochondrial genes.}, } @article {pmid24355929, year = {2014}, author = {Ma, WJ and Vavre, F and Beukeboom, LW}, title = {Manipulation of arthropod sex determination by endosymbionts: diversity and molecular mechanisms.}, journal = {Sexual development : genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation}, volume = {8}, number = {1-3}, pages = {59-73}, doi = {10.1159/000357024}, pmid = {24355929}, issn = {1661-5433}, mesh = {Animals ; Arthropods/*microbiology/*physiology ; *Biodiversity ; Haploidy ; Parthenogenesis/physiology ; Sex Determination Processes/physiology ; *Symbiosis ; }, abstract = {Arthropods exhibit a large variety of sex determination systems both at the chromosomal and molecular level. Male heterogamety, female heterogamety, and haplodiploidy occur frequently, but partially different genes are involved. Endosymbionts, such as Wolbachia, Cardinium,Rickettsia, and Spiroplasma, can manipulate host reproduction and sex determination. Four major reproductive manipulation types are distinguished: cytoplasmic incompatibility, thelytokous parthenogenesis, male killing, and feminization. In this review, the effects of these manipulation types and how they interfere with arthropod sex determination in terms of host developmental timing, alteration of sex determination, and modification of sexual differentiation pathways are summarized. Transitions between different manipulation types occur frequently which suggests that they are based on similar molecular processes. It is also discussed how mechanisms of reproductive manipulation and host sex determination can be informative on each other, with a special focus on haplodiploidy. Future directions on how the study of endosymbiotic manipulation of host reproduction can be key to further studies of arthropod sex determination are shown.}, } @article {pmid24351659, year = {2013}, author = {Hussain, M and O'Neill, SL and Asgari, S}, title = {Wolbachia interferes with the intracellular distribution of Argonaute 1 in the dengue vector Aedes aegypti by manipulating the host microRNAs.}, journal = {RNA biology}, volume = {10}, number = {12}, pages = {1868-1875}, pmid = {24351659}, issn = {1555-8584}, mesh = {Aedes/metabolism/*microbiology ; Animals ; Argonaute Proteins/*physiology ; Cell Line ; Cell Nucleus/*physiology ; Epigenesis, Genetic ; Gene Expression Regulation ; Insect Proteins/*physiology ; MicroRNAs/*genetics/metabolism ; Protein Stability ; Wolbachia/*physiology ; beta Karyopherins/*physiology ; }, abstract = {Argonaute proteins (AGOs) are vital components of the RNA-induced silencing complex in gene silencing. AGOs are indispensable for microRNA (miRNA) biogenesis as well as function, and are intracellularly localized to both the cytoplasm and the nucleus. Cytoplasmic AGO-miRNA complexes are mainly involved in cleavage or translational repression of target mRNAs while the nuclear ones are engaged in transcriptional gene silencing, methylation, chromatin remodeling, and splicing. In insects, AGO1 and AGO2 are involved in RNA interference and miRNA pathways but the components involved in their trafficking between the nucleus and the cytoplasm are not known. In this study, we found that importin β-4 facilitates AGO1 distribution to the nucleus, which is regulated by aae-miR-981 miRNA. The results also revealed association of prohibitin with AGO1 that may play an important role in its stability. Importantly, we found that AGO1 distribution to the nucleus is blocked by Wolbachia, an endosymbiotic bacterium introduced into the Dengue vector, Aedes aegypti. Our results provide basic mechanisms on intracellular trafficking of AGO1 in insects and how this may be altered by Wolbachia, which may affect trafficking of miRNAs to the nucleus leading to alteration in epigenetic effects.}, } @article {pmid24348259, year = {2013}, author = {Chrostek, E and Marialva, MS and Esteves, SS and Weinert, LA and Martinez, J and Jiggins, FM and Teixeira, L}, title = {Wolbachia variants induce differential protection to viruses in Drosophila melanogaster: a phenotypic and phylogenomic analysis.}, journal = {PLoS genetics}, volume = {9}, number = {12}, pages = {e1003896}, pmid = {24348259}, issn = {1553-7404}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology/virology ; Evolution, Molecular ; Genome, Insect ; Genomics ; Insect Viruses/genetics/pathogenicity ; Longevity/*genetics ; Phenotype ; Phylogeny ; Virus Diseases/*genetics ; Wolbachia/*genetics/growth & development ; }, abstract = {Wolbachia are intracellular bacterial symbionts that are able to protect various insect hosts from viral infections. This tripartite interaction was initially described in Drosophila melanogaster carrying wMel, its natural Wolbachia strain. wMel has been shown to be genetically polymorphic and there has been a recent change in variant frequencies in natural populations. We have compared the antiviral protection conferred by different wMel variants, their titres and influence on host longevity, in a genetically identical D. melanogaster host. The phenotypes cluster the variants into two groups--wMelCS-like and wMel-like. wMelCS-like variants give stronger protection against Drosophila C virus and Flock House virus, reach higher titres and often shorten the host lifespan. We have sequenced and assembled the genomes of these Wolbachia, and shown that the two phenotypic groups are two monophyletic groups. We have also analysed a virulent and over-replicating variant, wMelPop, which protects D. melanogaster even better than the closely related wMelCS. We have found that a ~21 kb region of the genome, encoding eight genes, is amplified seven times in wMelPop and may be the cause of its phenotypes. Our results indicate that the more protective wMelCS-like variants, which sometimes have a cost, were replaced by the less protective but more benign wMel-like variants. This has resulted in a recent reduction in virus resistance in D. melanogaster in natural populations worldwide. Our work helps to understand the natural variation in wMel and its evolutionary dynamics, and inform the use of Wolbachia in arthropod-borne disease control.}, } @article {pmid24347564, year = {2015}, author = {Goodacre, SL and Fricke, C and Martin, OY}, title = {A screen for bacterial endosymbionts in the model organisms Tribolium castaneum, T. confusum, Callosobruchus maculatus, and related species.}, journal = {Insect science}, volume = {22}, number = {2}, pages = {165-177}, doi = {10.1111/1744-7917.12096}, pmid = {24347564}, issn = {1744-7917}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Coleoptera/*microbiology ; Reproduction ; Rickettsia/genetics/*isolation & purification ; Spiroplasma/genetics/*isolation & purification ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Reproductive parasites such as Wolbachia are extremely widespread amongst the arthropods and can have a large influence over the reproduction and fitness of their hosts. Undetected infections could thus confound the results of a wide range of studies that focus on aspects of host behavior, reproduction, fitness, and degrees of reproductive isolation. This potential problem has already been underlined by work investigating the incidence of Wolbachia infections in stocks of the model system Drosophila melanogaster. Here we survey a range of lab stocks of further commonly used model arthropods, focusing especially on the flour beetles Tribolium castaneum and Tribolium confusum, the cowpea weevil Callosobruchus maculatus and related species (Coleoptera: Tenebrionidae and Bruchidae). These species are widespread stored product pests so knowledge of infections with symbionts further has potential use in informing biocontrol measures. Beetles were assessed for infection with 3 known microbial reproductive parasites: Wolbachia, Rickettsia, Spiroplasma. Infections with some of these microbes were found in some of the lab stocks studied, although overall infections were relatively rare. The consequences of finding infections in these or other species and the type of previous studies likely to be affected most are discussed.}, } @article {pmid24345405, year = {2014}, author = {Le Clec'h, W and Raimond, M and Bouchon, D and Sicard, M}, title = {Strength of the pathogenicity caused by feminizing Wolbachia after transfer in a new host: strain or dose effect?.}, journal = {Journal of invertebrate pathology}, volume = {116}, number = {}, pages = {18-26}, doi = {10.1016/j.jip.2013.12.003}, pmid = {24345405}, issn = {1096-0805}, mesh = {Animals ; Female ; *Host-Pathogen Interactions ; Isopoda/*microbiology ; Sex Determination Processes ; Virulence ; Wolbachia/*pathogenicity/physiology ; }, abstract = {The alphaproteobacteria Wolbachia pipientis are among the most common and widespread symbionts in the animal world. Their vertical transmission mode is predicted to favour genotypes with low virulence. On the contrary, horizontal transfers of Wolbachia from one host to another have been shown to possibly increase the symbiont virulence. This situation has been previously described when two feminizing Wolbachia strains, wVulC and wVulM, from the ovaries of the woodlouse Armadillidium vulgare were introduced into another woodlouse named Porcellio dilatatus. These two Wolbachia strains induced severe symptoms and eventually caused the death of the recipient host. However, symptoms and death appeared sooner with wVulC than with wVulM. To know whether this difference was due to variation in the dose of infection or a difference in virulence between the two Wolbachia strains, we performed controlled and gradual doses of injection with wVulC and wVulM in P. dilatatus. We showed that the two strains differed intrinsically in their virulence against P. dilatatus and that their virulence is related to the injection dose. Moreover, we showed that wVulC reached higher concentrations in the recipient host than wVulM suggesting a potential link between the bacterial titers and the levels of virulence. We also addressed the impact of the tissue source of the Wolbachia used for the transinfection and demonstrated that Wolbachia transinfected via hemolymph colonized the body of the recipient more quickly and caused accelerated symptoms compared to Wolbachia introduced via a crushed ovaries suspension.}, } @article {pmid24343914, year = {2014}, author = {Rainey, SM and Shah, P and Kohl, A and Dietrich, I}, title = {Understanding the Wolbachia-mediated inhibition of arboviruses in mosquitoes: progress and challenges.}, journal = {The Journal of general virology}, volume = {95}, number = {Pt 3}, pages = {517-530}, doi = {10.1099/vir.0.057422-0}, pmid = {24343914}, issn = {1465-2099}, support = {MC_UP_A550_1031/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 094664/Z/10/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Arboviruses/*physiology ; *Communicable Disease Control ; Communicable Diseases/virology ; Humans ; Insect Vectors/microbiology/physiology/*virology ; Wolbachia/*physiology ; }, abstract = {Arthropod-borne viruses (arboviruses) pose a considerable threat to human and animal health, yet effective control measures have proven difficult to implement, and novel means of controlling their replication in arthropod vectors, such as mosquitoes, are urgently required. One of the most exciting approaches to emerge from research on arthropods is the use of the endosymbiotic intracellular bacterium Wolbachia to control arbovirus transmission from mosquito to vertebrate. These α-proteobacteria propagate through insects, in part through modulation of host reproduction, thus ensuring spread through species and maintenance in nature. Since it was discovered that Wolbachia endosymbiosis inhibits insect virus replication in Drosophila species, these bacteria have also been shown to inhibit arbovirus replication and spread in mosquitoes. Importantly, it is not clear how these antiviral effects are mediated. This review will summarize recent work and discuss determinants of antiviral effectiveness that may differ between individual Wolbachia/vector/arbovirus interactions. We will also discuss the application of this approach to field settings and the associated risks.}, } @article {pmid24340120, year = {2013}, author = {Chu, BK and Deming, M and Biritwum, NK and Bougma, WR and Dorkenoo, AM and El-Setouhy, M and Fischer, PU and Gass, K and Gonzalez de Peña, M and Mercado-Hernandez, L and Kyelem, D and Lammie, PJ and Flueckiger, RM and Mwingira, UJ and Noordin, R and Offei Owusu, I and Ottesen, EA and Pavluck, A and Pilotte, N and Rao, RU and Samarasekera, D and Schmaedick, MA and Settinayake, S and Simonsen, PE and Supali, T and Taleo, F and Torres, M and Weil, GJ and Won, KY}, title = {Transmission assessment surveys (TAS) to define endpoints for lymphatic filariasis mass drug administration: a multicenter evaluation.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {12}, pages = {e2584}, pmid = {24340120}, issn = {1935-2735}, mesh = {Animals ; Brugia/isolation & purification ; Child ; Disease Transmission, Infectious/*prevention & control ; Elephantiasis, Filarial/*drug therapy/epidemiology/prevention & control/*transmission ; Epidemiological Monitoring ; Female ; Filaricides/*therapeutic use ; Humans ; Male ; Wolbachia/isolation & purification ; }, abstract = {BACKGROUND: Lymphatic filariasis (LF) is targeted for global elimination through treatment of entire at-risk populations with repeated annual mass drug administration (MDA). Essential for program success is defining and confirming the appropriate endpoint for MDA when transmission is presumed to have reached a level low enough that it cannot be sustained even in the absence of drug intervention. Guidelines advanced by WHO call for a transmission assessment survey (TAS) to determine if MDA can be stopped within an LF evaluation unit (EU) after at least five effective rounds of annual treatment. To test the value and practicality of these guidelines, a multicenter operational research trial was undertaken in 11 countries covering various geographic and epidemiological settings.

METHODOLOGY: The TAS was conducted twice in each EU with TAS-1 and TAS-2 approximately 24 months apart. Lot quality assurance sampling (LQAS) formed the basis of the TAS survey design but specific EU characteristics defined the survey site (school or community), eligible population (6-7 year olds or 1(st)-2(nd) graders), survey type (systematic or cluster-sampling), target sample size, and critical cutoff (a statistically powered threshold below which transmission is expected to be no longer sustainable). The primary diagnostic tools were the immunochromatographic (ICT) test for W. bancrofti EUs and the BmR1 test (Brugia Rapid or PanLF) for Brugia spp. EUs.

In 10 of 11 EUs, the number of TAS-1 positive cases was below the critical cutoff, indicating that MDA could be stopped. The same results were found in the follow-up TAS-2, therefore, confirming the previous decision outcome. Sample sizes were highly sex and age-representative and closely matched the target value after factoring in estimates of non-participation. The TAS was determined to be a practical and effective evaluation tool for stopping MDA although its validity for longer-term post-MDA surveillance requires further investigation.}, } @article {pmid24337178, year = {2014}, author = {Watanabe, M and Yukuhiro, F and Maeda, T and Miura, K and Kageyama, D}, title = {Novel strain of Spiroplasma found in flower bugs of the genus Orius (Hemiptera: Anthocoridae): transovarial transmission, coexistence with Wolbachia and varied population density.}, journal = {Microbial ecology}, volume = {67}, number = {1}, pages = {219-228}, pmid = {24337178}, issn = {1432-184X}, mesh = {Animals ; DNA, Bacterial/genetics ; Female ; Hemiptera/*microbiology ; Japan ; Male ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Spiroplasma/*classification/growth & development/isolation & purification ; Symbiosis ; Wolbachia/*growth & development ; }, abstract = {Spiroplasma, a group of small, wall-less, helical, and motile bacteria belonging to the Mollicutes, contains species with diverse life histories. To date, all the Spiroplasma strains that are known to be transmitted vertically in arthropod lineages belong to either the Spiroplasma ixodetis group or the Spiroplasma poulsonii group. Here, we found that a unique strain of Spiroplasma vertically transmitted in predatory flower bugs of the genus Orius belongs to the Spiroplasma insolitum group, which is a group of bacteria phylogenetically closely related to S. insolitum derived from the tickseed sunflower, Bidens sp. (Asterales: Asteraceae). The infection frequencies in natural populations were16.0% in Orius sauteri (n = 75), 40.5% in Orius nagaii (n = 37), and 8.0% in Orius minutus (n = 87). Orius strigicollis was not infected with Spiroplasma (n = 147). In the early stage of oogenesis (i.e., within the germarium), a large number of bacteria with the typical morphology of Spiroplasma existed, keeping a distance from Wolbachia bacteria. The Spiroplasma population seemed to increase during host development but Wolbachia population did not.}, } @article {pmid24337107, year = {2014}, author = {Caragata, EP and Rancès, E and O'Neill, SL and McGraw, EA}, title = {Competition for amino acids between Wolbachia and the mosquito host, Aedes aegypti.}, journal = {Microbial ecology}, volume = {67}, number = {1}, pages = {205-218}, pmid = {24337107}, issn = {1432-184X}, mesh = {Aedes/*microbiology/*physiology ; Amino Acids/*metabolism ; Animals ; Cholesterol/metabolism ; Female ; Fertility ; Humans ; Ovum/physiology ; Rats ; Rats, Inbred Lew ; Sheep ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The endosymbiont Wolbachia represents a promising method of dengue control, as it reduces the ability of the primary vector, the mosquito Aedes aegypti, to transmit viruses. When mosquitoes infected with the virulent Wolbachia strain wMelPop are fed non-human blood, there is a drastic reduction in mosquito fecundity and egg viability. Wolbachia has a reduced genome and is clearly dependent on its host for a wide range of nutritional needs. The fitness defects seen in wMelPop-infected A. aegypti could be explained by competition between the mosquito and the symbiont for essential blood meal nutrients, the profiles of which are suboptimal in non-human blood. Here, we examine cholesterol and amino acids as candidate molecules for competition, as they have critical roles in egg structural development and are known to vary between blood sources. We found that Wolbachia infection reduces total cholesterol levels in mosquitoes by 15-25%. We then showed that cholesterol supplementation of a rat blood meal did not improve fecundity or egg viability deficits. Conversely, amino acid supplementation of sucrose before and after a sheep blood meal led to statistically significant increases in fecundity of approximately 15-20 eggs per female and egg viability of 30-40%. This mosquito system provides the first empirical evidence of competition between Wolbachia and a host over amino acids and may suggest a general feature of Wolbachia-insect associations. These competitive processes could affect many aspects of host physiology and potentially mosquito fitness, a key concern for Wolbachia-based mosquito biocontrol.}, } @article {pmid24329998, year = {2014}, author = {Hughes, GL and Rasgon, JL}, title = {Transinfection: a method to investigate Wolbachia-host interactions and control arthropod-borne disease.}, journal = {Insect molecular biology}, volume = {23}, number = {2}, pages = {141-151}, pmid = {24329998}, issn = {1365-2583}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21 AI088311/AI/NIAID NIH HHS/United States ; R21AI088311/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Host-Pathogen Interactions ; Insecta/*microbiology ; Isopoda/*microbiology ; Pest Control, Biological/*methods ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {The bacterial endosymbiont Wolbachia manipulates arthropod host biology in numerous ways, including sex ratio distortion and differential offspring survival. These bacteria infect a vast array of arthropods, some of which pose serious agricultural and human health threats. Wolbachia-mediated phenotypes such as cytoplasmic incompatibility and/or pathogen interference can be used for vector and disease control; however, many medically important vectors and important agricultural species are uninfected or are infected with strains of Wolbachia that do not elicit phenotypes desirable for disease or pest control. The ability to transfer strains of Wolbachia into new hosts (transinfection) can create novel Wolbachia-host associations. Transinfection has two primary benefits. First, Wolbachia-host interactions can be examined to tease apart the influence of the host and bacteria on phenotypes. Second, desirable phenotypes induced by Wolbachia in a particular insect can be transferred to another recipient host. This can allow the manipulation of insect populations that transmit pathogens or detrimentally affect agriculture. As such, transinfection is a valuable tool to explore Wolbachia biology and control arthropod-borne disease. The present review summarizes what is currently known about Wolbachia transinfection methods and applications. We also provide a comprehensive list of published successful and unsuccessful Wolbachia transinfection attempts.}, } @article {pmid24324814, year = {2013}, author = {Valette, V and Bitome Essono, PY and Le Clec'h, W and Johnson, M and Bech, N and Grandjean, F}, title = {Multi-infections of feminizing Wolbachia strains in natural populations of the terrestrial isopod Armadillidium vulgare.}, journal = {PloS one}, volume = {8}, number = {12}, pages = {e82633}, pmid = {24324814}, issn = {1932-6203}, mesh = {Animals ; Female ; France ; Genotype ; Host-Pathogen Interactions ; Isopoda/*microbiology ; Male ; Polymerase Chain Reaction ; Prevalence ; Reproducibility of Results ; Rickettsiaceae Infections/veterinary ; Wolbachia/*genetics ; }, abstract = {Maternally inherited Wolbachia (α-Proteobacteria) are widespread parasitic reproductive manipulators. A growing number of studies have described the presence of different Wolbachia strains within a same host. To date, no naturally occurring multiple infections have been recorded in terrestrial isopods. This is true for Armadillidium vulgare which is known to harbor non simultaneously three Wolbachia strains. Traditionally, such Wolbachia are detected by PCR amplification of the wsp gene and strains are characterized by sequencing. The presence of nucleotide deletions or insertions within the wsp gene, among these three different strains, provides the opportunity to test a novel genotyping method. Herein, we designed a new primer pair able to amplify products whose lengths are specific to each Wolbachia strain so as to detect the presence of multi-infections in A. vulgare. Experimental injections of Wolbachia strains in Wolbachia-free females were used to validate the methodology. We re-investigated, using this novel method, the infection status of 40 females sampled in 2003 and previously described as mono-infected based on the classical sequencing method. Among these females, 29 were identified as bi-infected. It is the first time that naturally occurring multiple infections of Wolbachia are detected within an individual A. vulgare host. Additionally, we resampled 6 of these populations in 2010 to check the infection status of females.}, } @article {pmid24308996, year = {2014}, author = {Oliva, CF and Damiens, D and Benedict, MQ}, title = {Male reproductive biology of Aedes mosquitoes.}, journal = {Acta tropica}, volume = {132 Suppl}, number = {}, pages = {S12-9}, doi = {10.1016/j.actatropica.2013.11.021}, pmid = {24308996}, issn = {1873-6254}, mesh = {Aedes/*physiology ; Animals ; Infertility ; Male ; Mosquito Control/methods ; Pest Control, Biological/methods ; *Sexual Behavior, Animal ; }, abstract = {Among Aedes mosquitoes are species responsible for transmission of serious pathogens to humans. To cope with the current threats to long-term effectiveness of the traditional vector control methods, non-conventional control strategies are being developed. These include autocidal control such as the release of sterile males (sterile insect technique) and the release of Wolbachia-infected males to induce sexual sterility (incompatible insect technique) and pathogen-refractory strain replacement variations using Wolbachia. Sterile male types of techniques particularly depend on released males' ability to successfully mate with wild females. For that reason, a good understanding of male mating biology, including a thorough understanding of the reproductive system and mating capacity, increases the likelihood of success of such genetic vector control programmes. Here we review the literature concerning the reproduction of Aedes mosquitoes with an emphasis on the male biology. We consider sexual maturation, mate finding, insemination, male reproductive capacity, and the occurrence of multiple matings. We also discuss which parameters are of greatest importance for the successful implementation of autocidal control methods and propose questions for future research.}, } @article {pmid24282596, year = {2013}, author = {Walden, PM and Halili, MA and Archbold, JK and Lindahl, F and Fairlie, DP and Inaba, K and Martin, JL}, title = {The α-proteobacteria Wolbachia pipientis protein disulfide machinery has a regulatory mechanism absent in γ-proteobacteria.}, journal = {PloS one}, volume = {8}, number = {11}, pages = {e81440}, pmid = {24282596}, issn = {1932-6203}, support = {//Medical Research Council/United Kingdom ; }, mesh = {Amino Acid Sequence ; Bacterial Proteins/chemistry/*metabolism ; Base Sequence ; Catalysis ; DNA Primers ; Disulfides/*metabolism ; Gammaproteobacteria/*metabolism ; Molecular Sequence Data ; Oxidation-Reduction ; Sequence Homology, Amino Acid ; Wolbachia/*metabolism ; }, abstract = {The α-proteobacterium Wolbachia pipientis infects more than 65% of insect species worldwide and manipulates the host reproductive machinery to enable its own survival. It can live in mutualistic relationships with hosts that cause human disease, including mosquitoes that carry the Dengue virus. Like many other bacteria, Wolbachia contains disulfide bond forming (Dsb) proteins that introduce disulfide bonds into secreted effector proteins. The genome of the Wolbachia strain wMel encodes two DsbA-like proteins sharing just 21% sequence identity to each other, α-DsbA1 and α-DsbA2, and an integral membrane protein, α-DsbB. α-DsbA1 and α-DsbA2 both have a Cys-X-X-Cys active site that, by analogy with Escherichia coli DsbA, would need to be oxidized to the disulfide form to serve as a disulfide bond donor toward substrate proteins. Here we show that the integral membrane protein α-DsbB oxidizes α-DsbA1, but not α-DsbA2. The interaction between α-DsbA1 and α-DsbB is very specific, involving four essential cysteines located in the two periplasmic loops of α-DsbB. In the electron flow cascade, oxidation of α-DsbA1 by α-DsbB is initiated by an oxidizing quinone cofactor that interacts with the cysteine pair in the first periplasmic loop. Oxidizing power is transferred to the second cysteine pair, which directly interacts with α-DsbA1. This reaction is inhibited by a non-catalytic disulfide present in α-DsbA1, conserved in other α-proteobacterial DsbAs but not in γ-proteobacterial DsbAs. This is the first characterization of the integral membrane protein α-DsbB from Wolbachia and reveals that the non-catalytic cysteines of α-DsbA1 regulate the redox relay system in cooperation with α-DsbB.}, } @article {pmid24281548, year = {2014}, author = {Xie, J and Butler, S and Sanchez, G and Mateos, M}, title = {Male killing Spiroplasma protects Drosophila melanogaster against two parasitoid wasps.}, journal = {Heredity}, volume = {112}, number = {4}, pages = {399-408}, pmid = {24281548}, issn = {1365-2540}, mesh = {Animals ; Biological Evolution ; Cytoplasm/microbiology ; Defense Mechanisms ; Drosophila melanogaster/*microbiology/*parasitology ; Female ; Male ; Spiroplasma/*physiology ; Survival Analysis ; *Symbiosis ; Wasps/microbiology/*physiology ; Wolbachia ; }, abstract = {Maternally transmitted associations between endosymbiotic bacteria and insects are diverse and widespread in nature. Owing to imperfect vertical transmission, many heritable microbes have evolved compensational mechanisms to enhance their persistence in host lineages, such as manipulating host reproduction and conferring fitness benefits to host. Symbiont-mediated defense against natural enemies of hosts is increasingly recognized as an important mechanism by which endosymbionts enhance host fitness. Members of the genus Spiroplasma associated with distantly related Drosophila hosts are known to engage in either reproductive parasitism (i.e., male killing) or defense against natural enemies (the parasitic wasp Leptopilina heterotoma and a nematode). A male-killing strain of Spiroplasma (strain Melanogaster Sex Ratio Organism (MSRO)) co-occurs with Wolbachia (strain wMel) in certain wild populations of the model organism Drosophila melanogaster. We examined the effects of Spiroplasma MSRO and Wolbachia wMel on Drosophila survival against parasitism by two common wasps, Leptopilina heterotoma and Leptopilina boulardi, that differ in their host ranges and host evasion strategies. The results indicate that Spiroplasma MSRO prevents successful development of both wasps, and confers a small, albeit significant, increase in larva-to-adult survival of flies subjected to wasp attacks. We modeled the conditions under which defense can contribute to Spiroplasma persistence. Wolbachia also confers a weak, but significant, survival advantage to flies attacked by L. heterotoma. The host protective effects exhibited by Spiroplasma and Wolbachia are additive and may provide the conditions for such cotransmitted symbionts to become mutualists. Occurrence of Spiroplasma-mediated protection against distinct parasitoids in divergent Drosophila hosts suggests a general protection mechanism.}, } @article {pmid24274471, year = {2014}, author = {Hamilton, PT and Leong, JS and Koop, BF and Perlman, SJ}, title = {Transcriptional responses in a Drosophila defensive symbiosis.}, journal = {Molecular ecology}, volume = {23}, number = {6}, pages = {1558-1570}, doi = {10.1111/mec.12603}, pmid = {24274471}, issn = {1365-294X}, mesh = {Animals ; Bacterial Toxins/metabolism ; Drosophila/genetics/*microbiology/*parasitology ; Gene Expression Regulation ; Genes, Bacterial ; Nematoda/*pathogenicity ; Spiroplasma/*physiology ; *Symbiosis ; Transcriptome ; }, abstract = {Inherited symbionts are ubiquitous in insects and can have important consequences for the fitness of their hosts. Many inherited symbionts defend their hosts against parasites or other natural enemies; however, the means by which most symbionts confer protection is virtually unknown. We examine the mechanisms of defence in a recently discovered case of symbiont-mediated protection, where the bacterial symbiont Spiroplasma defends the fly Drosophila neotestacea from a virulent nematode parasite, Howardula aoronymphium. Using quantitative PCR of Spiroplasma infection intensities and whole transcriptome sequencing, we attempt to distinguish between the following modes of defence: symbiont-parasite competition, host immune priming and the production of toxic factors by Spiroplasma. Our findings do not support a model of exploitative competition between Howardula and Spiroplasma to mediate defence, nor do we find strong support for host immune priming during Spiroplasma infection. Interestingly, we recovered sequence for putative toxins encoded by Spiroplasma, including a novel putative ribosome-inactivating protein, transcripts of which are up-regulated in response to nematode exposure. Protection via the production of toxins may be a widely used and important mechanism in heritable defensive symbioses in insects.}, } @article {pmid24272923, year = {2014}, author = {Parrella, G and Nappo, AG and Manco, E and Greco, B and Giorgini, M}, title = {Invasion of the Q2 mitochondrial variant of Mediterranean Bemisia tabaci in southern Italy: possible role of bacterial endosymbionts.}, journal = {Pest management science}, volume = {70}, number = {10}, pages = {1514-1523}, doi = {10.1002/ps.3686}, pmid = {24272923}, issn = {1526-4998}, mesh = {Animals ; Bacteria/*genetics/*isolation & purification ; Female ; Genetic Variation ; Hemiptera/*genetics/*microbiology ; Introduced Species ; Italy ; Male ; Mitochondria/*genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; *Sex Ratio ; *Symbiosis ; }, abstract = {BACKGROUND: The whitefly Bemisia tabaci (Gennadius) is a complex of cryptic species, some of which, namely the Mediterranean (MED) and the Middle East-Asia Minor 1 (MEAM1), are highly invasive and injurious crop pests worldwide and able to displace local genotypes. Invasiveness of B. tabaci may depend on the phenotype of inherited bacterial endosymbionts. Here, the B. tabaci genetic diversity variation that has occurred in recent years in southern Italy was examined. Whitefly was genotyped by restriction fragment length polymorphism analysis of polymerase-chain-reaction-amplified fragments (PCR-RFLP) of the COI gene and molecular identification of endosymbionts. Possible factors leading to the observed genetic diversity were examined.

RESULTS: Q1 and Q2 mitochondrial types of MED, the only species found, coexisted in the field, while MEAM1 disappeared. A large spreading of Q2 (70% of individuals) was observed for the first time in Italy. Q2 showed a significant female-biased sex ratio and largely outnumbered Q1 on solanaceous hosts, in greenhouses and on insecticide-treated plants. Q1, with an even sex ratio, slightly prevailed on non-solanaceous hosts, especially on wild and untreated plants. Endosymbiont composition was associated with the mitochondrial type. Hamiltonella and Rickettsia were found at near fixation in Q1 and Q2 respectively; Arsenophonus, Cardinium and Wolbachia were found in both types, although at different frequencies.

CONCLUSIONS: Q2 invasion seems to have been favoured by the agroecological conditions of southern Italy and by the female-biased sex ratio. Endosymbionts may have a role in Q2 invasiveness, acting as sex-ratio manipulators (e.g. Rickettsia) and possibly by benefiting the host fitness.}, } @article {pmid24266987, year = {2014}, author = {Heintzman, PD and Elias, SA and Moore, K and Paszkiewicz, K and Barnes, I}, title = {Characterizing DNA preservation in degraded specimens of Amara alpina (Carabidae: Coleoptera).}, journal = {Molecular ecology resources}, volume = {14}, number = {3}, pages = {606-615}, doi = {10.1111/1755-0998.12205}, pmid = {24266987}, issn = {1755-0998}, mesh = {Animals ; Coleoptera/classification/*genetics ; DNA/*genetics ; DNA Contamination ; DNA, Mitochondrial/genetics ; Molecular Sequence Data ; Museums ; Phylogeny ; Preservation, Biological ; Sequence Analysis, DNA ; }, abstract = {DNA preserved in degraded beetle (Coleoptera) specimens, including those derived from dry-stored museum and ancient permafrost-preserved environments, could provide a valuable resource for researchers interested in species and population histories over timescales from decades to millenia. However, the potential of these samples as genetic resources is currently unassessed. Here, using Sanger and Illumina shotgun sequence data, we explored DNA preservation in specimens of the ground beetle Amara alpina, from both museum and ancient environments. Nearly all museum specimens had amplifiable DNA, with the maximum amplifiable fragment length decreasing with age. Amplification of DNA was only possible in 45% of ancient specimens. Preserved mitochondrial DNA fragments were significantly longer than those of nuclear DNA in both museum and ancient specimens. Metagenomic characterization of extracted DNA demonstrated that parasite-derived sequences, including Wolbachia and Spiroplasma, are recoverable from museum beetle specimens. Ancient DNA extracts contained beetle DNA in amounts comparable to museum specimens. Overall, our data demonstrate that there is great potential for both museum and ancient specimens of beetles in future genetic studies, and we see no reason why this would not be the case for other orders of insect.}, } @article {pmid24260298, year = {2013}, author = {Johansson, H and Dhaygude, K and Lindström, S and Helanterä, H and Sundström, L and Trontti, K}, title = {A metatranscriptomic approach to the identification of microbiota associated with the ant Formica exsecta.}, journal = {PloS one}, volume = {8}, number = {11}, pages = {e79777}, pmid = {24260298}, issn = {1932-6203}, mesh = {Animals ; Ants/*genetics/*microbiology ; Behavior, Animal ; Microbiota/*genetics ; Phylogeny ; RNA, Ribosomal/genetics ; Sequence Analysis, DNA/methods ; Social Behavior ; Symbiosis/genetics ; Transcriptome/*genetics ; }, abstract = {Social insects live in cooperative colonies, often in high densities and with closely related individuals, and interact using social contact behaviours. Compared to solitary insects, social insects have evolved multi-level immunity that includes immune responses common to holometabolous insects, and social immunity, which is exclusive to social taxa. This suggests that social insects may be subject to high pathogen pressure, yet relatively little is known about the range of symbiotic and pathogenic microbial communities that associate with social insects. In this study we examined transcriptome data generated from the ant Formica exsecta for sequences identifying as microbes (or other organisms potentially of non-ant origin). Sequences showing homology to two viruses and several other potentially or obligate intracellular organisms, such as Wolbachia, Arsenophonus, Entomoplasmatales and Microsporidia, were present in the transcriptome data. These homologous sequence matches correspond to genera/species that have previously been associated with a variety of insects, including social insects. There were also sequences with identity to several other microbes such as common moulds and soil bacteria. We conclude that this sequence data provides a starting point for a deeper understanding of the biological interactions between a species of ant and the micro- and macrobiotic communities that it potentially encounters.}, } @article {pmid24254373, year = {2014}, author = {Niranjan Reddy, B and Gupta, B and Rao, BP}, title = {Vector population manipulation for control of arboviruses--a novel prospect for India.}, journal = {Pest management science}, volume = {70}, number = {4}, pages = {517-523}, doi = {10.1002/ps.3676}, pmid = {24254373}, issn = {1526-4998}, mesh = {Aedes/*genetics/microbiology ; Animals ; *Arboviruses ; Chikungunya Fever/prevention & control/transmission ; Chikungunya virus/genetics ; Dengue/prevention & control/*transmission ; Dengue Virus/genetics ; Humans ; India ; *Insect Vectors ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; Risk Assessment ; Transgenes ; Wolbachia ; }, abstract = {India, the seventh largest country in the world, has diverse geographical and climatic regions with vast rural and peri-urban areas. Many are experiencing an escalation in the spread and intensity of numerous human diseases transmitted by insects. Classically, the management of these vector-borne diseases is underpinned by either chemical insecticides and/or environmental management targeted at the vector. However, these methods or their present implementation do not offer acceptable levels of control, and more effective and sustainable options are now available. Genetic strategies for the prevention of arbovirus transmission are most advanced for dengue and chikungunya, targeting their primary vector, Aedes aegypti. The national burden in terms of morbidity and mortality as a direct consequence of dengue virus in India is considered to be the largest worldwide, over 4 times that of any other country. Presently, new genetic technologies are undergoing field evaluation of their biosafety and efficacy in several countries. This paper discusses the merits of these approaches and argues for fair and transparent appraisal in India as a matter of urgency. Identification of any associated risks and their appropriate mitigation are fundamental to that process.}, } @article {pmid24252486, year = {2014}, author = {Bourtzis, K and Dobson, SL and Xi, Z and Rasgon, JL and Calvitti, M and Moreira, LA and Bossin, HC and Moretti, R and Baton, LA and Hughes, GL and Mavingui, P and Gilles, JR}, title = {Harnessing mosquito-Wolbachia symbiosis for vector and disease control.}, journal = {Acta tropica}, volume = {132 Suppl}, number = {}, pages = {S150-63}, doi = {10.1016/j.actatropica.2013.11.004}, pmid = {24252486}, issn = {1873-6254}, mesh = {Aedes/*growth & development/*microbiology ; Animals ; *Insect Vectors ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; *Symbiosis ; Wolbachia/growth & development/*physiology ; }, abstract = {Mosquito species, members of the genera Aedes, Anopheles and Culex, are the major vectors of human pathogens including protozoa (Plasmodium sp.), filariae and of a variety of viruses (causing dengue, chikungunya, yellow fever, West Nile). There is lack of efficient methods and tools to treat many of the diseases caused by these major human pathogens, since no efficient vaccines or drugs are available; even in malaria where insecticide use and drug therapies have reduced incidence, 219 million cases still occurred in 2010. Therefore efforts are currently focused on the control of vector populations. Insecticides alone are insufficient to control mosquito populations since reduced susceptibility and even resistance is being observed more and more frequently. There is also increased concern about the toxic effects of insecticides on non-target (even beneficial) insect populations, on humans and the environment. During recent years, the role of symbionts in the biology, ecology and evolution of insect species has been well-documented and has led to suggestions that they could potentially be used as tools to control pests and therefore diseases. Wolbachia is perhaps the most renowned insect symbiont, mainly due to its ability to manipulate insect reproduction and to interfere with major human pathogens thus providing new avenues for pest control. We herein present recent achievements in the field of mosquito-Wolbachia symbiosis with an emphasis on Aedes albopictus. We also discuss how Wolbachia symbiosis can be harnessed for vector control as well as the potential to combine the sterile insect technique and Wolbachia-based approaches for the enhancement of population suppression programs.}, } @article {pmid24250820, year = {2013}, author = {Yang, XH and Zhu, DH and Liu, Z and Zhao, L and Su, CY}, title = {High levels of multiple infections, recombination and horizontal transmission of Wolbachia in the Andricus mukaigawae (Hymenoptera; Cynipidae) communities.}, journal = {PloS one}, volume = {8}, number = {11}, pages = {e78970}, pmid = {24250820}, issn = {1932-6203}, mesh = {Animals ; DNA, Mitochondrial/genetics ; Disease Transmission, Infectious ; Haplotypes ; Host-Pathogen Interactions/*genetics ; Hymenoptera/genetics/*microbiology ; Phylogeny ; *Recombination, Genetic ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria of arthropods and nematodes. In arthropods, they manipulate the reproduction of their hosts to facilitate their own spread in host populations, causing cytoplasmic incompatibility, parthenogenesis induction, feminization of genetic males and male-killing. In this study, we investigated Wolbachia infection and studied wsp (Wolbachia surface protein) sequences in three wasp species associated with the unisexual galls of A. mukaigawae with the aim of determining the transmission mode and the reason for multiple infections of Wolbachia. Frequency of Wolbachia infected populations for A. mukaigawae, Synergus japonicus (inquiline), and Torymus sp. (parasitoid) was 75%, 100%, and 100%, respectively. Multiple Wolbachia infections were detected in A. mukaigawae and S. japonicus, with 5 and 8 Wolbachia strains, respectively. The two host species shared 5 Wolbachia strains and were infected by identical strains in several locations, indicating horizontal transmission of Wolbachia. The transmission potentially takes place through gall tissues, which the larvae of both wasps feed on. Furthermore, three recombination events of Wolbachia were observed: the strains W8, W2 and W6 apparently have derived from W3 and W5a, W6 and W7, W4 and W9, respectively. W8 and W2 and their respective parental strains were detected in S. japonicus. W6 was detected with only one parent (W4) in S. japonicus; W9 was detected in Torymus sp., suggesting horizontal transmission between hosts and parasitoids. In conclusion, our research supports earlier studies that horizontal transmission of Wolbachia, a symbiont of the Rickettsiales order, may be plant-mediated or take place between hosts and parasitoids. Our research provides novel molecular evidence for multiple recombination events of Wolbachia in gall wasp communities. We suggest that genomic recombination and potential plant-mediated horizontal transmission may be attributable to the high levels of multiple Wolbachia infections observed in A. mukaigawae and S. japonicus.}, } @article {pmid24247133, year = {2014}, author = {Bah, GS and Ward, EL and Srivastava, A and Trees, AJ and Tanya, VN and Makepeace, BL}, title = {Efficacy of three-week oxytetracycline or rifampin monotherapy compared with a combination regimen against the filarial nematode Onchocerca ochengi.}, journal = {Antimicrobial agents and chemotherapy}, volume = {58}, number = {2}, pages = {801-810}, pmid = {24247133}, issn = {1098-6596}, mesh = {Adult ; Animals ; Anti-Bacterial Agents/*pharmacology ; Cattle ; Drug Administration Schedule ; Drug Combinations ; Humans ; Microbial Sensitivity Tests ; Microbial Viability/drug effects ; Onchocerca/drug effects/microbiology/physiology ; Onchocerciasis/*drug therapy/microbiology/parasitology/*veterinary ; Oxytetracycline/antagonists & inhibitors/*pharmacology ; Parasite Load/veterinary ; Rifampin/*pharmacology ; Skin/drug effects/parasitology ; Symbiosis ; Treatment Outcome ; Wolbachia/*drug effects/physiology ; }, abstract = {Onchocerciasis (river blindness), caused by the filarial nematode Onchocerca volvulus, is a major cause of visual impairment and dermatitis in sub-Saharan Africa. As O. volvulus contains an obligatory bacterial symbiont (Wolbachia), it is susceptible to antibiotic chemotherapy, although current regimens are considered too prolonged for community-level control programs. The aim of this study was to compare the efficacies of oxytetracycline and rifampin, administered separately or in combination, against a close relative of O. volvulus (Onchocerca ochengi) in cattle. Six animals per group were treated with continuous or intermittent oxytetracycline regimens, and effects on adult worm viability, dermal microfilarial loads, and Wolbachia density in worm tissues were assessed. Subsequently, the efficacies of 3-week regimens of oxytetracycline and rifampin alone and a combination regimen were compared, and rifampin levels in plasma and skin were quantified. A 6-month regimen of oxytetracycline with monthly dosing was strongly adulticidal, while 3-week and 6-week regimens exhibited weaker adulticidal effects. However, all three regimens achieved >2-log reductions in microfilarial load. In contrast, rifampin monotherapy and oxytetracycline-rifampin duotherapy failed to induce substantive reductions in either adult worm burden or microfilarial load, although a borderline effect on Wolbachia density was observed following duotherapy. Dermal rifampin levels were maintained above the MIC for >24 h after a single intravenous dose. We conclude that oxytetracycline-rifampin duotherapy is less efficacious against O. ochengi than oxytetracycline alone. Further studies will be required to determine whether rifampin reduces oxytetracycline bioavailability in this system, as suggested by human studies using other tetracycline-rifampin combinations.}, } @article {pmid24242251, year = {2014}, author = {Newell, PD and Douglas, AE}, title = {Interspecies interactions determine the impact of the gut microbiota on nutrient allocation in Drosophila melanogaster.}, journal = {Applied and environmental microbiology}, volume = {80}, number = {2}, pages = {788-796}, pmid = {24242251}, issn = {1098-5336}, support = {F32 GM099374/GM/NIGMS NIH HHS/United States ; R01 GM095372/GM/NIGMS NIH HHS/United States ; 1R01GM095372/GM/NIGMS NIH HHS/United States ; F32GM099374/GM/NIGMS NIH HHS/United States ; }, mesh = {Acetobacter/metabolism ; Animal Nutritional Physiological Phenomena ; Animals ; Drosophila melanogaster/growth & development/*microbiology ; Female ; Gastrointestinal Tract/*microbiology ; Lactobacillus/metabolism ; *Microbiota ; Phenotype ; Triglycerides/*metabolism ; Wolbachia ; }, abstract = {The animal gut is perpetually exposed to microorganisms, and this microbiota affects development, nutrient allocation, and immune homeostasis. A major challenge is to understand the contribution of individual microbial species and interactions among species in shaping these microbe-dependent traits. Using the Drosophila melanogaster gut microbiota, we tested whether microbe-dependent performance and nutritional traits of Drosophila are functionally modular, i.e., whether the impact of each microbial taxon on host traits is independent of the presence of other microbial taxa. Gnotobiotic flies were constructed with one or a set of five of the Acetobacter and Lactobacillus species which dominate the gut microbiota of conventional flies (Drosophila with untreated microbiota). Axenic (microbiota-free) flies exhibited prolonged development time and elevated glucose and triglyceride contents. The low glucose content of conventional flies was recapitulated in gnotobiotic Drosophila flies colonized with any of the 5 bacterial taxa tested. In contrast, the development rates and triglyceride levels in monocolonized flies varied depending on the taxon present: Acetobacter species supported the largest reductions, while most Lactobacillus species had no effect. Only flies with both Acetobacter and Lactobacillus had triglyceride contents restored to the level in conventional flies. This could be attributed to two processes: Lactobacillus-mediated promotion of Acetobacter abundance in the fly and a significant negative correlation between fly triglyceride content and Acetobacter abundance. We conclude that the microbial basis of host traits varies in both specificity and modularity; microbe-mediated reduction in glucose is relatively nonspecific and modular, while triglyceride content is influenced by interactions among microbes.}, } @article {pmid24241712, year = {2014}, author = {Johnston, KL and Ford, L and Taylor, MJ}, title = {Overcoming the challenges of drug discovery for neglected tropical diseases: the A·WOL experience.}, journal = {Journal of biomolecular screening}, volume = {19}, number = {3}, pages = {335-343}, doi = {10.1177/1087057113511270}, pmid = {24241712}, issn = {1552-454X}, mesh = {*Drug Discovery ; Elephantiasis, Filarial/diagnosis/drug therapy/prevention & control ; Global Health ; Humans ; Neglected Diseases/diagnosis/*drug therapy/*prevention & control ; Onchocerciasis/diagnosis/drug therapy/prevention & control ; *Tropical Medicine ; Wolbachia/drug effects ; World Health Organization ; }, abstract = {Neglected tropical diseases (NTDs) are a group of 17 diseases that typically affect poor people in tropical countries. Each has been neglected for decades in terms of funding, research, and policy, but the recent grouping of them into one unit, which can be targeted using integrated control measures, together with increased advocacy has helped to place them on the global health agenda. The World Health Organization has set ambitious goals to control or eliminate 10 NTDs by 2020 and launched a roadmap in January 2012 to guide this global plan. The result of the launch meeting, which brought together representatives from the pharmaceutical industry, donors, and politicians, was the London Declaration: a series of commitments to provide more drugs, research, and funds to achieve the 2020 goals. Drug discovery and development for these diseases are extremely challenging, and this article highlights these challenges in the context of the London Declaration, before focusing on an example of a drug discovery and development program for the NTDs onchocerciasis and lymphatic filariasis (the anti-Wolbachia consortium, A·WOL).}, } @article {pmid24223136, year = {2013}, author = {Ritter, S and Michalski, SG and Settele, J and Wiemers, M and Fric, ZF and Sielezniew, M and Šašić, M and Rozier, Y and Durka, W}, title = {Wolbachia infections mimic cryptic speciation in two parasitic butterfly species, Phengaris teleius and P. nausithous (Lepidoptera: Lycaenidae).}, journal = {PloS one}, volume = {8}, number = {11}, pages = {e78107}, pmid = {24223136}, issn = {1932-6203}, mesh = {Animals ; Ants/parasitology ; Asia ; Butterflies/classification/*genetics ; Cell Nucleus/genetics ; Electron Transport Complex IV/classification/*genetics ; Europe ; Female ; Genetic Markers ; *Genetic Speciation ; Genetic Variation ; *Genome, Mitochondrial ; Insect Proteins/classification/*genetics ; Microsatellite Repeats ; Phylogeny ; Phylogeography ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Deep mitochondrial divergence within species may result from cryptic speciation, from phylogeographic isolation or from endosymbiotic bacteria like Wolbachia that manipulate host reproduction. Phengaris butterflies are social parasites that spend most of their life in close relationship with ants. Previously, cryptic speciation has been hypothesised for two Phengaris species based on divergent mtDNA sequences. Since Phengaris species are highly endangered, the existence of cryptic species would have drastic consequences for conservation and management. We tested for cryptic speciation and alternative scenarios in P. teleius and P. nausithous based on a comprehensive sample across their Palaearctic ranges using COI gene sequences, nuclear microsatellites and tests for Wolbachia. In both species a deep mitochondrial split occurring 0.65-1.97 myrs ago was observed that did not correspond with microsatellite data but was concordant with Wolbachia infection. Haplotypes previously attributed to cryptic species were part of the Wolbachia-infected clades. In both species remaining phylogeographic structure was largely consistent between mitochondrial and nuclear genomes. In P. teleius several mitochondrial and nuclear groups were observed in East Asia while a single haplogroup and nuclear cluster prevailed across continental Eurasia. Neutrality tests suggested rapid demographic expansion into that area. In contrast, P. nausithous had several mitochondrial and nuclear groups in Europe, suggesting a complex phylogeographic history in the western part of the species range. We conclude that deep intraspecific divergences found in DNA barcode studies do not necessarily need to represent cryptic speciation but instead can be due to both infection by Wolbachia and phylogeographic structure.}, } @article {pmid24221888, year = {2014}, author = {Zhang, D and Zhan, X and Wu, X and Yang, X and Liang, G and Zheng, Z and Li, Z and Wu, Y and Zheng, X}, title = {A field survey for Wolbchia and phage WO infections of Aedes albopictus in Guangzhou City, China.}, journal = {Parasitology research}, volume = {113}, number = {1}, pages = {399-404}, pmid = {24221888}, issn = {1432-1955}, mesh = {Aedes/*microbiology/*virology ; Animals ; Bacteriophages/genetics/*isolation & purification ; China ; DNA Primers ; Female ; Insect Vectors/microbiology/virology ; Male ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/genetics/*isolation & purification/virology ; }, abstract = {Wolbachia are maternal endosymbiotic bacterium, which infect a diverse range of arthropods, ranging from 20 to 76% in nature. They are capable of inducing a wide range of reproductive abnormalities to their hosts, such as cytoplasmic incompatibility (CI), which has been proposed to be used as a tool to modify mosquitoes that are resistant to the development of pathogen, as an alternative vector control strategy. Here, we evaluated the prevalence of Wolbachia and phage WO infections in the field population of Aedes albopictus in Guangzhou City via polymerase chain reaction (PCR) assay using the Wolbachia specific Wolbachia surface protein (wsp) and phage WO orf7 gene primers. Based on the results of PCR and phylogeny analysis, we found that A. albopictus in Guangzhou City were infected with two Wolbachia strains, wAlbA and wAlbB. Phage WO, the virus-infected Wolbachia, was also detected in A. albopictus. One hundred and ten female individuals were screened via PCR, with 109 super-infected with Wolbachia and one sample single-infected with wAlbB strain. And 104 of 113 male individuals were both infected with wAlbA and wAlbB, and nine male samples were found to be infected with wAlbA strain only. The infection rates of phage WO in female and male individuals were 82.73 and 46.02%, respectively. These results showed that the natural Wolbachia and phage WO infections in A. albopictus population in Guangzhou were at a higher frequency at present, indicating that Wolbachia appear to be a better candidate nature resource for biological control insect vectors to reduce vector-borne diseases.}, } @article {pmid24216442, year = {2013}, author = {Zhang, YK and Ding, XL and Zhang, KJ and Hong, XY}, title = {Wolbachia play an important role in affecting mtDNA variation of Tetranychus truncatus (Trombidiformes: Tetranychidae).}, journal = {Environmental entomology}, volume = {42}, number = {6}, pages = {1240-1245}, doi = {10.1603/EN13085}, pmid = {24216442}, issn = {1938-2936}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Female ; Genetic Variation ; Haplotypes ; Phylogeny ; Tetranychidae/genetics/*microbiology ; Wolbachia/*genetics/physiology ; }, abstract = {The prevalence of the endosymbiont Wolbachia and its effects on mitochondria variation were analyzed in seven natural populations of Tetranychus truncatus Ehara (Trombidiformes: Tetranychidae) in current study. Five Wolbachia strains (wtru1, wtru5, wtru7, wtru8, and wtru12) were detected based on the surface protein of Wolbachia (wsp) sequence data and the multiple locus sequences typing data, suggesting that multiple separate invasions have occurred. Part of mitochondrial cytochrome oxidase subunit I gene was sequenced from infected individuals revealing 10 different haplotypes. As predicted, the haplotype and nucleotide diversity were lower in infected individuals than that in uninfected individuals. Furthermore, phylogenetic and analysis of molecular variance analyses revealed that the distribution of mtDNA haplotypes is not associated with geography. Rather, it is strongly concordant with infection status. These data support the hypothesis that Wolbachia infection can affect the genetic structure and diversity of the host mites.}, } @article {pmid24214960, year = {2013}, author = {Amir, A and Zeisel, A and Zuk, O and Elgart, M and Stern, S and Shamir, O and Turnbaugh, PJ and Soen, Y and Shental, N}, title = {High-resolution microbial community reconstruction by integrating short reads from multiple 16S rRNA regions.}, journal = {Nucleic acids research}, volume = {41}, number = {22}, pages = {e205}, pmid = {24214960}, issn = {1362-4962}, support = {P50 GM068763/GM/NIGMS NIH HHS/United States ; }, mesh = {Acetobacter/genetics/isolation & purification ; Animals ; Bacteria/*classification/genetics/isolation & purification ; Drosophila melanogaster/microbiology ; High-Throughput Nucleotide Sequencing/*methods ; Humans ; Models, Statistical ; *Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Saliva/microbiology ; Sequence Analysis, DNA/*methods ; Wolbachia/genetics/isolation & purification ; }, abstract = {The emergence of massively parallel sequencing technology has revolutionized microbial profiling, allowing the unprecedented comparison of microbial diversity across time and space in a wide range of host-associated and environmental ecosystems. Although the high-throughput nature of such methods enables the detection of low-frequency bacteria, these advances come at the cost of sequencing read length, limiting the phylogenetic resolution possible by current methods. Here, we present a generic approach for integrating short reads from large genomic regions, thus enabling phylogenetic resolution far exceeding current methods. The approach is based on a mapping to a statistical model that is later solved as a constrained optimization problem. We demonstrate the utility of this method by analyzing human saliva and Drosophila samples, using Illumina single-end sequencing of a 750 bp amplicon of the 16S rRNA gene. Phylogenetic resolution is significantly extended while reducing the number of falsely detected bacteria, as compared with standard single-region Roche 454 Pyrosequencing. Our approach can be seamlessly applied to simultaneous sequencing of multiple genes providing a higher resolution view of the composition and activity of complex microbial communities.}, } @article {pmid24204251, year = {2013}, author = {Pinto, SB and Stainton, K and Harris, S and Kambris, Z and Sutton, ER and Bonsall, MB and Parkhill, J and Sinkins, SP}, title = {Transcriptional regulation of Culex pipiens mosquitoes by Wolbachia influences cytoplasmic incompatibility.}, journal = {PLoS pathogens}, volume = {9}, number = {10}, pages = {e1003647}, pmid = {24204251}, issn = {1553-7374}, support = {//Wellcome Trust/United Kingdom ; 076964/WT_/Wellcome Trust/United Kingdom ; 079059/WT_/Wellcome Trust/United Kingdom ; 095121/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Culex/genetics/metabolism/microbiology ; Drosophila melanogaster ; Female ; Gene Knockdown Techniques ; Genome, Bacterial/physiology ; *Infertility, Female/genetics/metabolism/microbiology ; *Insect Proteins/biosynthesis/genetics ; Male ; Meiosis/genetics ; *Transcription Factors/biosynthesis/genetics ; *Up-Regulation ; *Wolbachia/genetics/metabolism ; Zinc Fingers/genetics ; }, abstract = {Cytoplasmic incompatibility (CI) induced by the endosymbiont Wolbachia pipientis causes complex patterns of crossing sterility between populations of the Culex pipiens group of mosquitoes. The molecular basis of the phenotype is yet to be defined. In order to investigate what host changes may underlie CI at the molecular level, we examined the transcription of a homolog of the Drosophila melanogaster gene grauzone that encodes a zinc finger protein and acts as a regulator of female meiosis, in which mutations can cause sterility. Upregulation was observed in Wolbachia-infected C. pipiens group individuals relative to Wolbachia-cured lines and the level of upregulation differed between lines that were reproductively incompatible. Knockdown analysis of this gene using RNAi showed an effect on hatch rates in a Wolbachia infected Culex molestus line. Furthermore, in later stages of development an effect on developmental progression in CI embryos occurs in bidirectionally incompatible crosses. The genome of a wPip Wolbachia strain variant from Culex molestus was sequenced and compared with the genome of a wPip variant with which it was incompatible. Three genes in inserted or deleted regions were newly identified in the C. molestus wPip genome, one of which is a transcriptional regulator labelled wtrM. When this gene was transfected into adult Culex mosquitoes, upregulation of the grauzone homolog was observed. These data suggest that Wolbachia-mediated regulation of host gene expression is a component of the mechanism of cytoplasmic incompatibility.}, } @article {pmid24195062, year = {2013}, author = {Wang, J and Weiss, BL and Aksoy, S}, title = {Tsetse fly microbiota: form and function.}, journal = {Frontiers in cellular and infection microbiology}, volume = {3}, number = {}, pages = {69}, pmid = {24195062}, issn = {2235-2988}, support = {R01 AI051584/AI/NIAID NIH HHS/United States ; AI051584/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Host-Parasite Interactions ; *Microbiota ; *Symbiosis ; Tsetse Flies/*microbiology/*physiology ; }, abstract = {Tsetse flies are the primary vectors of African trypanosomes, which cause Human and Animal African trypanosomiasis in 36 countries in sub-Saharan Africa. These flies have also established symbiotic associations with bacterial and viral microorganisms. Laboratory-reared tsetse flies harbor up to four vertically transmitted organisms-obligate Wigglesworthia, commensal Sodalis, parasitic Wolbachia and Salivary Gland Hypertrophy Virus (SGHV). Field-captured tsetse can harbor these symbionts as well as environmentally acquired commensal bacteria. This microbial community influences several aspects of tsetse's physiology, including nutrition, fecundity and vector competence. This review provides a detailed description of tsetse's microbiome, and describes the physiology underlying host-microbe, and microbe-microbe, interactions that occur in this fly.}, } @article {pmid24190075, year = {2013}, author = {Woolfit, M and Iturbe-Ormaetxe, I and Brownlie, JC and Walker, T and Riegler, M and Seleznev, A and Popovici, J and Rancès, E and Wee, BA and Pavlides, J and Sullivan, MJ and Beatson, SA and Lane, A and Sidhu, M and McMeniman, CJ and McGraw, EA and O'Neill, SL}, title = {Genomic evolution of the pathogenic Wolbachia strain, wMelPop.}, journal = {Genome biology and evolution}, volume = {5}, number = {11}, pages = {2189-2204}, pmid = {24190075}, issn = {1759-6653}, mesh = {Aedes/microbiology ; Animals ; Base Sequence ; Cell Line ; Drosophila melanogaster/microbiology ; *Evolution, Molecular ; Gene Duplication ; *Genome, Bacterial ; Molecular Sequence Data ; Mutation ; Selection, Genetic ; Virulence/genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Most strains of the widespread endosymbiotic bacterium Wolbachia pipientis are benign or behave as reproductive parasites. The pathogenic strain wMelPop is a striking exception, however: it overreplicates in its insect hosts and causes severe life shortening. The mechanism of this pathogenesis is currently unknown. We have sequenced the genomes of three variants of wMelPop and of the closely related nonpathogenic strain wMelCS. We show that the genomes of wMelCS and wMelPop appear to be identical in the nonrepeat regions of the genome and differ detectably only by the triplication of a 19-kb region that is unlikely to be associated with life shortening, demonstrating that dramatic differences in the host phenotype caused by this endosymbiont may be the result of only minor genetic changes. We also compare the genomes of the original wMelPop strain from Drosophila melanogaster and two sequential derivatives, wMelPop-CLA and wMelPop-PGYP. To develop wMelPop as a novel biocontrol agent, it was first transinfected into and passaged in mosquito cell lines for approximately 3.5 years, generating wMelPop-CLA. This cell line-passaged strain was then transinfected into Aedes aegypti mosquitoes, creating wMelPop-PGYP, which was sequenced after 4 years in the insect host. We observe a rapid burst of genomic changes during cell line passaging, but no further mutations were detected after transinfection into mosquitoes, indicating either that host preadaptation had occurred in cell lines, that cell lines are a more selectively permissive environment than animal hosts, or both. Our results provide valuable data on the rates of genomic and phenotypic change in Wolbachia associated with host shifts over short time scales.}, } @article {pmid24180121, year = {2013}, author = {Crain, PR and Crowley, PH and Dobson, SL}, title = {Wolbachia re-replacement without incompatibility: potential for intended and unintended consequences.}, journal = {Journal of medical entomology}, volume = {50}, number = {5}, pages = {1152-1158}, doi = {10.1603/me12263}, pmid = {24180121}, issn = {0022-2585}, support = {AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/*microbiology/*physiology ; Animal Distribution ; Animals ; Australia ; Female ; *Genetic Fitness ; Male ; Models, Biological ; Mosquito Control/*methods ; Population Dynamics ; Wolbachia/*physiology ; }, abstract = {Recent releases of mosquitoes infected with different Wolbachia types into Australia have led to conditions that have not been observed naturally and that have not been considered theoretically. In an ongoing public health campaign, two Wolbachia infection types have been introduced, and the infections differ in their fitness cost imposed on the host mosquito. The dynamics of each infection type as it interacts with the indigenous uninfected host population have been previously examined via models and empirical tests. Here we conduct a modeling analysis to predict the dynamics when the two infection types do not remain geographically isolated. The results demonstrate the potential replacement and loss of the more costly infection type, which may not be the desired public health outcome.}, } @article {pmid24180100, year = {2013}, author = {Hapairai, LK and Sang, MA and Sinkins, SP and Bossin, HC}, title = {Population studies of the filarial vector Aedes polynesiensis (Diptera: Culicidae) in two island settings of French Polynesia.}, journal = {Journal of medical entomology}, volume = {50}, number = {5}, pages = {965-976}, doi = {10.1603/me12246}, pmid = {24180100}, issn = {0022-2585}, support = {095121//Wellcome Trust/United Kingdom ; / WT095121/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*parasitology/*physiology ; Animal Distribution ; Animals ; Elephantiasis, Filarial/*transmission ; Female ; Filarioidea/*physiology ; Humans ; Male ; Polynesia ; Population Density ; Risk Assessment ; }, abstract = {A mark-release-recapture study was conducted to estimate the adult population size, migration, and dispersal patterns of male and female Aedes (Stegomyia) polynesiensis (Marks) in a valley of Moorea, a volcanic island, and a motu (islet) on the atoll of Tetiaroa, two settings typical of the Society Islands. Aedes polynesiensis recapture rate was high for females and low for males. The distribution of Aedes species in the valley was heterogeneous. Marked individuals dispersed to most parts of the motu and over great distances in the valley for some females. The study provides insights into the field dynamics of Ae. polynesiensis populations and confirms that more efficient sampling methods are warranted. There was no evidence of active migration between motus on the atoll, suggesting that Tetiaroa is a suitable site for small-scale initial open releases of Wolbachia incompatible insect technique and other sterile insect technique-like suppression or replacement strategies.}, } @article {pmid24167782, year = {2013}, author = {Newton, RR and Newton, IL}, title = {PhyBin: binning trees by topology.}, journal = {PeerJ}, volume = {1}, number = {}, pages = {e187}, pmid = {24167782}, issn = {2167-8359}, abstract = {A major goal of many evolutionary analyses is to determine the true evolutionary history of an organism. Molecular methods that rely on the phylogenetic signal generated by a few to a handful of loci can be used to approximate the evolution of the entire organism but fall short of providing a global, genome-wide, perspective on evolutionary processes. Indeed, individual genes in a genome may have different evolutionary histories. Therefore, it is informative to analyze the number and kind of phylogenetic topologies found within an orthologous set of genes across a genome. Here we present PhyBin: a flexible program for clustering gene trees based on topological structure. PhyBin can generate bins of topologies corresponding to exactly identical trees or can utilize Robinson-Fould's distance matrices to generate clusters of similar trees, using a user-defined threshold. Additionally, PhyBin allows the user to adjust for potential noise in the dataset (as may be produced when comparing very closely related organisms) by pre-processing trees to collapse very short branches or those nodes not meeting a defined bootstrap threshold. As a test case, we generated individual trees based on an orthologous gene set from 10 Wolbachia species across four different supergroups (A-D) and utilized PhyBin to categorize the complete set of topologies produced from this dataset. Using this approach, we were able to show that although a single topology generally dominated the analysis, confirming the separation of the supergroups, many genes supported alternative evolutionary histories. Because PhyBin's output provides the user with lists of gene trees in each topological cluster, it can be used to explore potential reasons for discrepancies between phylogenies including homoplasies, long-branch attraction, or horizontal gene transfer events.}, } @article {pmid24164708, year = {2014}, author = {Awrahman, ZA and Champion de Crespigny, F and Wedell, N}, title = {The impact of Wolbachia, male age and mating history on cytoplasmic incompatibility and sperm transfer in Drosophila simulans.}, journal = {Journal of evolutionary biology}, volume = {27}, number = {1}, pages = {1-10}, doi = {10.1111/jeb.12270}, pmid = {24164708}, issn = {1420-9101}, mesh = {Aging/physiology ; Animals ; *Copulation ; Drosophila simulans/microbiology/*physiology ; Female ; Male ; Reproductive Isolation ; Wolbachia/*physiology ; }, abstract = {Most insects harbour a variety of maternally inherited endosymbionts, the most widespread being Wolbachia pipientis that commonly induce cytoplasmic incompatibility (CI) and reduced hatching success in crosses between infected males and uninfected females. High temperature and increasing male age are known to reduce the level of CI in a variety of insects. In Drosophila simulans, infected males have been shown to mate at a higher rate than uninfected males. By examining the impact of mating rate independent of age, this study investigates whether a high mating rate confers an advantage to infected males through restoring their compatibility with uninfected females over and above the effect of age. The impact of Wolbachia infection, male mating rate and age on the number of sperm transferred to females during copulation and how it relates to CI expression was also assessed. As predicted, we found that reproductive compatibility was restored faster in males that mate at higher rate than that of low mating and virgin males, and that the effect of mating history was over and above the effect of male age. Nonvirgin infected males transferred fewer sperm than uninfected males during copulation, and mating at a high rate resulted in the transfer of fewer sperm per mating irrespective of infection status. These results indicate that the advantage to infected males of mating at a high rate is through restoration of reproductive compatibility with uninfected females, whereas uninfected males appear to trade off the number of sperm transferred per mating with female encounter rate and success in sperm competition. This study highlights the importance Wolbachia may play in sexual selection by affecting male reproductive strategies.}, } @article {pmid24152719, year = {2014}, author = {Darby, AC and Gill, AC and Armstrong, SD and Hartley, CS and Xia, D and Wastling, JM and Makepeace, BL}, title = {Integrated transcriptomic and proteomic analysis of the global response of Wolbachia to doxycycline-induced stress.}, journal = {The ISME journal}, volume = {8}, number = {4}, pages = {925-937}, pmid = {24152719}, issn = {1751-7370}, mesh = {ATP-Binding Cassette Transporters/genetics ; Adenosine Triphosphatases/genetics ; Animals ; Anti-Bacterial Agents/*pharmacology ; Arthropods/microbiology ; Bacterial Proteins/genetics ; Cell Line ; Doxycycline/*pharmacology ; Gene Expression Regulation, Bacterial/drug effects ; Membrane Transport Proteins/genetics ; *Proteomics ; RNA, Bacterial/genetics ; RNA, Untranslated ; Stress, Physiological/*genetics ; Wolbachia/*drug effects/*genetics/metabolism ; }, abstract = {The bacterium Wolbachia (order Rickettsiales), representing perhaps the most abundant vertically transmitted microbe worldwide, infects arthropods and filarial nematodes. In arthropods, Wolbachia can induce reproductive alterations and interfere with the transmission of several arthropod-borne pathogens. In addition, Wolbachia is an obligate mutualist of the filarial parasites that cause lymphatic filariasis and onchocerciasis in the tropics. Targeting Wolbachia with tetracycline antibiotics leads to sterilisation and ultimately death of adult filariae. However, several weeks of treatment are required, restricting the implementation of this control strategy. To date, the response of Wolbachia to stress has not been investigated, and almost nothing is known about global regulation of gene expression in this organism. We exposed an arthropod Wolbachia strain to doxycycline in vitro, and analysed differential expression by directional RNA-seq and label-free, quantitative proteomics. We found that Wolbachia responded not only by modulating expression of the translation machinery, but also by upregulating nucleotide synthesis and energy metabolism, while downregulating outer membrane proteins. Moreover, Wolbachia increased the expression of a key component of the twin-arginine translocase (tatA) and a phosphate ABC transporter ATPase (PstB); the latter is associated with decreased susceptibility to antimicrobials in free-living bacteria. Finally, the downregulation of 6S RNA during translational inhibition suggests that this small RNA is involved in growth rate control. Despite its highly reduced genome, Wolbachia shows a surprising ability to regulate gene expression during exposure to a potent stressor. Our findings have general relevance for the chemotherapy of obligate intracellular bacteria and the mechanistic basis of persistence in the Rickettsiales.}, } @article {pmid24146634, year = {2013}, author = {Robinson, KM and Sieber, KB and Dunning Hotopp, JC}, title = {A review of bacteria-animal lateral gene transfer may inform our understanding of diseases like cancer.}, journal = {PLoS genetics}, volume = {9}, number = {10}, pages = {e1003877}, pmid = {24146634}, issn = {1553-7404}, support = {DP2 OD007372/OD/NIH HHS/United States ; 1-DP2-OD007372/OD/NIH HHS/United States ; }, mesh = {Animals ; Chromosomes/genetics/microbiology ; DNA, Bacterial/*genetics ; Gene Transfer, Horizontal/*genetics ; Humans ; Interspersed Repetitive Sequences ; Neoplasms/*genetics/microbiology/virology ; Phylogeny ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Lateral gene transfer (LGT) from bacteria to animals occurs more frequently than was appreciated prior to the advent of genome sequencing. In 2007, LGT from bacterial Wolbachia endosymbionts was detected in ~33% of the sequenced arthropod genomes using a bioinformatic approach. Today, Wolbachia/host LGT is thought to be widespread and many other cases of bacteria-animal LGT have been described. In insects, LGT may be more frequently associated with endosymbionts that colonize germ cells and germ stem cells, like Wolbachia endosymbionts. We speculate that LGT may occur from bacteria to a wide variety of eukaryotes, but only becomes vertically inherited when it occurs in germ cells. As such, LGT may happen routinely in somatic cells but never become inherited or fixed in the population. Lack of inheritance of such mutations greatly decreases our ability to detect them. In this review, we propose that such noninherited bacterial DNA integration into chromosomes in human somatic cells could induce mutations leading to cancer or autoimmune diseases in a manner analogous to mobile elements and viral integrations.}, } @article {pmid24126721, year = {2013}, author = {Roberts, CH and Mongkolsapaya, J and Screaton, G}, title = {New opportunities for control of dengue virus.}, journal = {Current opinion in infectious diseases}, volume = {26}, number = {6}, pages = {567-574}, doi = {10.1097/QCO.0000000000000016}, pmid = {24126721}, issn = {1473-6527}, support = {095541/WT_/Wellcome Trust/United Kingdom ; G0400720/MRC_/Medical Research Council/United Kingdom ; G0801508/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Antiviral Agents/*therapeutic use ; Dengue/*prevention & control ; *Dengue Vaccines ; Humans ; Immunotherapy/*methods ; Mosquito Control/*methods ; }, abstract = {PURPOSE OF REVIEW: Dengue is a rapidly spreading vector-borne disease estimated to infect 400 million people worldwide. To date, there are no licensed treatments or vaccines. The last few years have seen significant developments in dengue control strategies. In this review, we will address four key areas: vaccines, vector control, antivirals and immunotherapeutics.

RECENT FINDINGS: The first generation of dengue vaccines is able to induce good serological responses in test individuals. However, the recent Sanofi-Pasteur trial in Thailand found that a good serological response did not correlate with clinical protection. This trial did not demonstrate an increase in cases of severe disease following immunization, suggesting that concerns over vaccine-related immune enhancement may have been overcome. The bacterium Wolbachia appears to control dengue proliferation in Aedes mosquitoes, and field studies are underway. A large number of antivirals are in early-stage development and may prove useful in epidemics. Monoclonal antibodies have been postulated to have a clinical role. Whether their clinical application is feasible has yet to be seen.

SUMMARY: Marked improvements in our knowledge of dengue have been made over the recent years. Sadly, clinical application remains some years away.}, } @article {pmid24125079, year = {2013}, author = {Segoli, M and Stouthamer, R and Stouthamer, CM and Rugman-Jones, P and Rosenheim, JA}, title = {The effect of Wolbachia on the lifetime reproductive success of its insect host in the field.}, journal = {Journal of evolutionary biology}, volume = {26}, number = {12}, pages = {2716-2720}, doi = {10.1111/jeb.12264}, pmid = {24125079}, issn = {1420-9101}, mesh = {Animals ; Insecta/*microbiology/physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a widespread endosymbiont that induces dramatic manipulations of its host's reproduction. Although there has been substantial progress in the developing theory for Wolbachia-host interactions and in measuring the effects of Wolbachia on host fitness in the laboratory, there is a widely recognized need to quantify the effects of Wolbachia on the host fitness in the field. The wasp Anagrus sophiae, an egg parasitoid of planthoppers, carries a Wolbachia strain that induces parthenogenesis, but its effects on the fitness of its Anagrus host are unknown. We developed a method to estimate the realized lifetime reproductive success of female wasps by collecting them soon after they die naturally in the field, counting the number of eggs remaining in their ovaries and quantifying Wolbachia density in their body. We sampled from a highly infected A. sophiae population and found no evidence for Wolbachia virulence and possible evidence for positive effects of Wolbachia on realized reproductive success.}, } @article {pmid24118435, year = {2013}, author = {Gerth, M and Röthe, J and Bleidorn, C}, title = {Tracing horizontal Wolbachia movements among bees (Anthophila): a combined approach using multilocus sequence typing data and host phylogeny.}, journal = {Molecular ecology}, volume = {22}, number = {24}, pages = {6149-6162}, doi = {10.1111/mec.12549}, pmid = {24118435}, issn = {1365-294X}, mesh = {Animals ; Bayes Theorem ; Bees/genetics/*microbiology ; *Biological Evolution ; *Gene Transfer, Horizontal ; Models, Genetic ; Molecular Sequence Data ; Multilocus Sequence Typing ; *Phylogeny ; Symbiosis/genetics ; Wolbachia/classification/*genetics ; }, abstract = {The endosymbiotic bacterium Wolbachia enhances its spread via vertical transmission by generating reproductive effects in its hosts, most notably cytoplasmic incompatibility (CI). Additionally, frequent interspecific horizontal transfer is evident from a lack of phylogenetic congruence between Wolbachia and its hosts. The mechanisms of this lateral transfer are largely unclear. To identify potential pathways of Wolbachia movements, we performed multilocus sequence typing of Wolbachia strains from bees (Anthophila). Using a host phylogeny and ecological data, we tested various models of horizontal endosymbiont transmission. In general, Wolbachia strains seem to be randomly distributed among bee hosts. Kleptoparasite-host associations among bees as well as other ecological links could not be supported as sole basis for the spread of Wolbachia. However, cophylogenetic analyses and divergence time estimations suggest that Wolbachia may persist within a host lineage over considerable timescales and that strictly vertical transmission and subsequent random loss of infections across lineages may have had a greater impact on Wolbachia strain distribution than previously estimated. Although general conclusions about Wolbachia movements among arthropod hosts cannot be made, we present a framework by which precise assumptions about shared evolutionary histories of Wolbachia and a host taxon can be modelled and tested.}, } @article {pmid24118111, year = {2013}, author = {Early, AM and Clark, AG}, title = {Monophyly of Wolbachia pipientis genomes within Drosophila melanogaster: geographic structuring, titre variation and host effects across five populations.}, journal = {Molecular ecology}, volume = {22}, number = {23}, pages = {5765-5778}, pmid = {24118111}, issn = {1365-294X}, support = {R01 AI064950/AI/NIAID NIH HHS/United States ; R01AI064950/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bayes Theorem ; DNA, Mitochondrial/genetics ; Drosophila melanogaster/genetics/*microbiology ; *Evolution, Molecular ; Genetics, Population ; *Genome, Bacterial ; Haplotypes ; Phenotype ; Phylogeny ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Wolbachia pipientis is one of the most widely studied endosymbionts today, yet we know little about its short-term adaptation and evolution. Here, using a set of 91 inbred Drosophila melanogaster lines from five populations, we explore patterns of diversity and recent evolution in the Wolbachia strain wMel. Within the D. melanogaster lines, we identify six major mitochondrial clades and four wMel clades. Concordant with past studies, the Wolbachia haplotypes contain an overall low level of nucleotide diversity, yet they still display geographic structuring. Using Bayesian analysis informed with demographic estimates of colonization times, we estimate that all extant D. melanogaster mitochondrial haplotypes coalesce to a Wolbachia-infected ancestor approximately 2200 years ago. Finally, we measure wMel titre within the infected flies and find that titre varies across populations, an effect attributable to host genetic factors. This demonstration of local phenotypic divergence suggests that intraspecific host genetic variation plays a key role in shaping this model symbiotic system.}, } @article {pmid24098543, year = {2013}, author = {Giraud, I and Valette, V and Bech, N and Grandjean, F and Cordaux, R}, title = {Isolation and characterization of microsatellite loci for the isopod crustacean Armadillidium vulgare and transferability in terrestrial isopods.}, journal = {PloS one}, volume = {8}, number = {10}, pages = {e76639}, pmid = {24098543}, issn = {1932-6203}, mesh = {Animals ; Biological Evolution ; Female ; Feminization/*genetics/microbiology ; Genetic Loci ; Genetic Variation ; Genomic Library ; Isopoda/*genetics/microbiology ; Male ; *Microsatellite Repeats ; Phylogeny ; *Sex Determination Processes ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Armadillidium vulgare is a terrestrial isopod (Crustacea, Oniscidea) which harbors Wolbachia bacterial endosymbionts. A. vulgare is the major model for the study of Wolbachia-mediated feminization of genetic males in crustaceans. As a consequence of their impact on host sex determination mechanisms, Wolbachia endosymbionts are thought to significantly influence A. vulgare evolution on various grounds, including population genetic structure, diversity and reproduction strategies. To provide molecular tools for examining these questions, we isolated microsatellite loci through 454 pyrosequencing of a repeat-enriched A. vulgare genomic library. We selected 14 markers and developed three polymorphic microsatellite multiplex kits. We tested the kits on two A. vulgare natural populations and found high genetic variation, thereby making it possible to investigate the impact of Wolbachia endosymbionts on A. vulgare nuclear variation at unprecedented resolution. In addition, we tested the transferability of these kits by cross-species amplification in five other terrestrial isopod species harboring Wolbachia endosymbionts. The microsatellite loci showed good transferability in particular in Armadillidium nasatum and Chaetophiloscia elongata, for which these markers represent promising tools for future genetic studies.}, } @article {pmid24080157, year = {2013}, author = {Wiwatanaratanabutr, I}, title = {Distribution, diversity and density of wolbachial infections in cladocerans and copepods from Thailand.}, journal = {Journal of invertebrate pathology}, volume = {114}, number = {3}, pages = {341-345}, doi = {10.1016/j.jip.2013.04.014}, pmid = {24080157}, issn = {1096-0805}, mesh = {Animals ; Biodiversity ; Cladocera/*microbiology ; Copepoda/*microbiology ; DNA, Bacterial/chemistry ; Fresh Water ; Geography ; Population Density ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA ; Species Specificity ; Thailand ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Species of the genus Wolbachia comprise a group of Rickettsia-like, maternally-inherited bacteria that cause several reproductive alterations in arthropod hosts. The best known are cytoplasmic incompatibility and feminization. Here, the first systematic surveys of wolbachial infections in cladocerans and copepods from six geographic regions of Thailand, including Northern, Northeastern, Western, Central, Eastern and Southern are reported. Using gene amplification assays with wsp and groE primers, wolbachiae were detected in 239 (4 spp.) of 1885 (57 spp.) copepods and cladocerans from all regions of Thailand surveyed. Screening results obtained with wsp primers or groE primers were similar in all cases. The presence of wolbachiae was only detected in copepods, not in cladocerans. Sex ratio analyses of the progeny of two species of copepods, Mesocyclops aspericornis and Mesocyclops thermocyclopoides, naturally or artificially infected with wolbachiae showed infection causes feminization (female-bias). The relative density if infection in naturally infected populations of three copepod species, M. thermocyclopoides, Heliodiaptomus elegans and Neodiaptomus blachei, were determined using real-time quantitative PCR assay based on the wsp gene. The density of wolbachiae in M. thermocyclopoides was significantly higher than in the other two species. These results suggest that wolbachial infections are distributed throughout Thailand, and that possibly the natural occurrence of these in copepods may be due to their predation on mosquito larvae. This apparent novel biology may have importance as a genetic drive system for control of vector borne diseases in the future.}, } @article {pmid24079981, year = {2014}, author = {Schäberle, TF and Schiefer, A and Schmitz, A and König, GM and Hoerauf, A and Pfarr, K}, title = {Corallopyronin A - a promising antibiotic for treatment of filariasis.}, journal = {International journal of medical microbiology : IJMM}, volume = {304}, number = {1}, pages = {72-78}, doi = {10.1016/j.ijmm.2013.08.010}, pmid = {24079981}, issn = {1618-0607}, mesh = {Animals ; Anti-Bacterial Agents/biosynthesis/isolation & purification/*pharmacology/therapeutic use ; Filariasis/*drug therapy ; Filaricides/isolation & purification/*pharmacology/therapeutic use ; Filarioidea/*microbiology/physiology ; Humans ; Lactones/isolation & purification/*pharmacology/therapeutic use ; Symbiosis ; Wolbachia/*drug effects/physiology ; }, abstract = {Lymphatic filariasis and onchocerciasis are diseases of severe morbidity that affect the poorest of the poor in the world. The diseases are caused by filarial nematodes that are transmitted by mosquitoes or biting blackflies and are endemic to more than 80 countries worldwide, mainly in the tropics and sub-tropics. Current control programs aim to eliminate the diseases by distributing antifilarial drugs. However, the primary effect of the drugs is to kill the microfilariae in the blood or skin, thus preventing uptake by the obligate insect vector. Since the adult worms live 10 years or longer, drug distribution requires many years of treatment, which is a heavy burden on the burgeoning health care systems. Sub-optimal response, possible resistance and inadequate population coverage lessen the chances for successful elimination in all endemic areas. The search for new drugs that could enhance elimination by permanently sterilizing or killing adult worms has identified the Wolbachia intracellular bacteria of filarial nematodes as a target. Depleting the obligate endosymbionts from the worms with doxycycline or rifampicin causes a permanent block in oogenesis, embryogenesis and development, and in slow death of the adult worms. These two antibiotics are suitable for individual drug administration, but caveats exist for their inclusion in broader drug administration programs. Here we review Wolbachia as targets for antifilarial drug discovery and highlight the natural product corallopyronin A as an effective drug that is currently being developed specifically for use against filarial nematodes.}, } @article {pmid24077701, year = {2013}, author = {Wang, GH and Xiao, JH and Xiong, TL and Li, Z and Murphy, RW and Huang, DW}, title = {High-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) for determination of a highly degenerated prophage WO genome in a Wolbachia strain infecting a fig wasp species.}, journal = {Applied and environmental microbiology}, volume = {79}, number = {23}, pages = {7476-7481}, pmid = {24077701}, issn = {1098-5336}, mesh = {Animals ; Bacteriophages/*genetics ; DNA Viruses/*genetics/isolation & purification ; DNA, Viral/*chemistry/*genetics ; Ficus/parasitology ; *Genome, Viral ; Molecular Sequence Data ; Polymerase Chain Reaction/methods ; Prophages/*genetics ; Sequence Analysis, DNA ; Wasps/microbiology ; Wolbachia/isolation & purification/*virology ; }, abstract = {Temperate bacteriophage WO is a model system for studying tripartite interactions among viruses, bacteria, and eukaryotes, especially investigations of the genomic stability of obligate intracellular bacteria. Few WO genomes exist because of the difficulty in isolating viral DNA from eukaryotic hosts, and most reports are by-products of Wolbachia sequencing. Only one partial genome of a WO phage has been determined directly from isolated particles. We determine the complete genome sequence of prophage WO (WOSol) in Wolbachia strain wSol, which infects the fig wasp Ceratosolen solmsi (Hymenoptera: Chalcidoidea), by high-efficiency thermal asymmetric interlaced PCR. The genome of WOSol is highly degenerated and disrupted by a large region (14,267 bp) from Wolbachia. Consistent with previous molecular studies of multiple WO genomes, the genome of WOSol appears to have evolved by single nucleotide mutations and recombinations.}, } @article {pmid24068927, year = {2013}, author = {Kriesner, P and Hoffmann, AA and Lee, SF and Turelli, M and Weeks, AR}, title = {Rapid sequential spread of two Wolbachia variants in Drosophila simulans.}, journal = {PLoS pathogens}, volume = {9}, number = {9}, pages = {e1003607}, pmid = {24068927}, issn = {1553-7374}, mesh = {Animals ; Australia ; Bacterial Proteins/chemistry/genetics/metabolism ; Base Sequence ; DNA, Bacterial/chemistry/metabolism ; DNA, Mitochondrial/chemistry/metabolism ; Drosophila/growth & development/*microbiology/physiology ; Embryo, Nonmammalian/*microbiology ; Female ; Fertility ; Male ; *Models, Biological ; Molecular Sequence Data ; Molecular Typing ; Mutation ; Pest Control, Biological ; Spatio-Temporal Analysis ; Species Specificity ; *Symbiosis ; Wolbachia/classification/isolation & purification/*physiology ; }, abstract = {The maternally inherited intracellular bacteria Wolbachia can manipulate host reproduction in various ways that foster frequency increases within and among host populations. Manipulations involving cytoplasmic incompatibility (CI), where matings between infected males and uninfected females produce non-viable embryos, are common in arthropods and produce a reproductive advantage for infected females. CI was associated with the spread of Wolbachia variant wRi in Californian populations of Drosophila simulans, which was interpreted as a bistable wave, in which local infection frequencies tend to increase only once the infection becomes sufficiently common to offset imperfect maternal transmission and infection costs. However, maternally inherited Wolbachia are expected to evolve towards mutualism, and they are known to increase host fitness by protecting against infectious microbes or increasing fecundity. We describe the sequential spread over approximately 20 years in natural populations of D. simulans on the east coast of Australia of two Wolbachia variants (wAu and wRi), only one of which causes significant CI, with wRi displacing wAu since 2004. Wolbachia and mtDNA frequency data and analyses suggest that these dynamics, as well as the earlier spread in California, are best understood as Fisherian waves of favourable variants, in which local spread tends to occur from arbitrarily low frequencies. We discuss implications for Wolbachia-host dynamics and coevolution and for applications of Wolbachia to disease control.}, } @article {pmid24059916, year = {2013}, author = {Sinkins, SP}, title = {Wolbachia and arbovirus inhibition in mosquitoes.}, journal = {Future microbiology}, volume = {8}, number = {10}, pages = {1249-1256}, doi = {10.2217/fmb.13.95}, pmid = {24059916}, issn = {1746-0921}, support = {095121//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/immunology/metabolism/*microbiology/*virology ; Animals ; Arboviruses/*physiology ; Dengue Virus/*physiology ; Lipid Metabolism ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a maternally inherited intracellular bacteria that can manipulate the reproduction of their insect hosts, and cytoplasmic incompatibility allows them to spread through mosquito populations. When particular strains of Wolbachia are transferred into certain Aedes mosquito species, the transmission capacity of important arthropod-borne viruses can be suppressed or abolished in laboratory challenges. Viral inhibition is associated with higher densities of transinfecting Wolbachia compared with wild-type strains of the bacterium. The upregulation of innate immune effectors can contribute to virus inhibition in Aedes aegypti, but does not seem to be required. Modulation of autophagy and lipid metabolism, and intracellular competition between viruses and bacteria for lipids, provide promising hypotheses for the mechanism of inhibition. Transinfecting virus-inhibiting strains can produce higher fitness costs than wild-type mosquito Wolbachia; however, this is not always the case, and the wMel strain has already been introduced to high frequency in wild Ae. aegypti populations.}, } @article {pmid24053607, year = {2013}, author = {Ioannidis, P and Johnston, KL and Riley, DR and Kumar, N and White, JR and Olarte, KT and Ott, S and Tallon, LJ and Foster, JM and Taylor, MJ and Dunning Hotopp, JC}, title = {Extensively duplicated and transcriptionally active recent lateral gene transfer from a bacterial Wolbachia endosymbiont to its host filarial nematode Brugia malayi.}, journal = {BMC genomics}, volume = {14}, number = {}, pages = {639}, pmid = {24053607}, issn = {1471-2164}, support = {DP2 OD007372/OD/NIH HHS/United States ; 1-DP2-OD007372/OD/NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*genetics ; DNA, Bacterial/genetics ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Genome, Helminth ; Open Reading Frames ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Lymphatic filariasis is a neglected tropical disease afflicting more than 120 million people, while another 1.3 billion people are at risk of infection. The nematode worm Brugia malayi is one of the causative agents of the disease and exists in a mutualistic symbiosis with Wolbachia bacteria. Since extensive lateral gene transfer occurs frequently between Wolbachia and its hosts, we sought to measure the extent of such LGT in B. malayi by whole genome sequencing of Wolbachia-depleted worms.

RESULTS: A considerable fraction (at least 115.4-kbp, or 10.6%) of the 1.08-Mbp Wolbachia wBm genome has been transferred to its nematode host and retains high levels of similarity, including 227 wBm genes and gene fragments. Complete open reading frames were transferred for 32 of these genes, meaning they have the potential to produce functional proteins. Moreover, four transfers have evidence of life stage-specific regulation of transcription at levels similar to other nematode transcripts, strengthening the possibility that they are functional.

CONCLUSIONS: There is extensive and ongoing transfer of Wolbachia DNA to the worm genome and some transfers are transcribed in a stage-specific manner at biologically relevant levels.}, } @article {pmid24044755, year = {2013}, author = {Nutman, TB}, title = {Insights into the pathogenesis of disease in human lymphatic filariasis.}, journal = {Lymphatic research and biology}, volume = {11}, number = {3}, pages = {144-148}, pmid = {24044755}, issn = {1557-8585}, support = {//Intramural NIH HHS/United States ; }, mesh = {Animals ; Elephantiasis, Filarial/*parasitology ; Filarioidea/microbiology/*physiology ; Host-Parasite Interactions ; Host-Pathogen Interactions ; Humans ; Lymph Nodes/*parasitology/pathology ; Lymphatic System/*parasitology/pathology ; Wolbachia/physiology ; }, abstract = {Although two thirds of the 120 million people infected with lymph-dwelling filarial parasites have subclinical infections, ∼40 million have lymphedema and/or other pathologic manifestations including hydroceles (and other forms of urogenital disease), episodic adenolymphangitis, lymphedema, and (in its most severe form) elephantiasis. Adult filarial worms reside in the lymphatics and lymph nodes and induce lymphatic dilatation. Progressive lymphatic damage and pathology results primarily from the host inflammatory response to the parasites but also perhaps from the host inflammatory response to the parasite's Wolbachia endosymbiont and as a consequence of superimposed bacterial or fungal infections. This review will attempt to shed light on disease pathogenesis in lymphatic filariasis.}, } @article {pmid24014528, year = {2013}, author = {Fink, C and Staubach, F and Kuenzel, S and Baines, JF and Roeder, T}, title = {Noninvasive analysis of microbiome dynamics in the fruit fly Drosophila melanogaster.}, journal = {Applied and environmental microbiology}, volume = {79}, number = {22}, pages = {6984-6988}, pmid = {24014528}, issn = {1098-5336}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; DNA, Bacterial/genetics ; Drosophila melanogaster/*microbiology ; Feces/microbiology ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/isolation & purification ; }, abstract = {The diversity and structure of the intestinal microbial community has a strong influence on life history. To understand how hosts and microbes interact, model organisms with comparatively simple microbial communities, such as the fruit fly (Drosophila melanogaster), offer key advantages. However, studies of the Drosophila microbiome are limited to a single point in time, because flies are typically sacrificed for DNA extraction. In order to test whether noninvasive approaches, such as sampling of fly feces, could be a means to assess fly-associated communities over time on the same cohort of flies, we compared the microbial communities of fly feces, dissected fly intestines, and whole flies across three different Drosophila strains. Bacterial species identified in either whole flies or isolated intestines were reproducibly found in feces samples. Although the bacterial communities of feces and intestinal samples were not identical, they shared similarities and obviously the same origin. In contrast to material from whole flies and intestines, feces samples were not compromised by Wolbachia spp. infections, which are widespread in laboratory and wild strains. In a proof-of-principle experiment, we showed that simple nutritional interventions, such as a high-fat diet or short-term starvation, had drastic and long-lasting effects on the micobiome. Thus, the analysis of feces can supplement the toolbox for microbiome studies in Drosophila, unleashing the full potential of such studies in time course experiments where multiple samples from single populations are obtained during aging, development, or experimental manipulations.}, } @article {pmid24013349, year = {2013}, author = {Scheitz, CJ and Guo, Y and Early, AM and Harshman, LG and Clark, AG}, title = {Heritability and inter-population differences in lipid profiles of Drosophila melanogaster.}, journal = {PloS one}, volume = {8}, number = {8}, pages = {e72726}, pmid = {24013349}, issn = {1932-6203}, support = {P20 RR016475/RR/NCRR NIH HHS/United States ; R01 DK074136/DK/NIDDK NIH HHS/United States ; P20 RR16475/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster ; Fatty Acids/*metabolism ; Female ; Lipid Metabolism/*physiology ; Male ; Membrane Fluidity/*physiology ; Species Specificity ; Wolbachia/metabolism ; }, abstract = {Characterizing and understanding the complex spectrum of lipids in higher organisms lags far behind our analysis of genome and transcriptome sequences. Here we generate and evaluate comprehensive lipid profiles (>200 lipids) of 92 inbred lines from five different Drosophila melanogaster populations. We find that the majority of lipid species are highly heritable, and even lipids with odd-chain fatty acids, which cannot be generated by the fly itself, also have high heritabilities. Abundance of the endosymbiont Wolbachia, a potential provider of odd-chained lipids, was positively correlated with this group of lipids. Additionally, we show that despite years of laboratory rearing on the same medium, the lipid profiles of the five geographic populations are sufficiently distinct for population discrimination. Our data predicts a strikingly different membrane fluidity for flies from the Netherlands, which is supported by their increased ethanol tolerance. We find that 18% of lipids show strong concentration differences between males and females. Through an analysis of the correlation structure of the lipid classes, we find modules of co-regulated lipids and begin to associate these with metabolic constraints. Our data provide a foundation for developing associations between variation in lipid composition with variation in other metabolic attributes, with genome-wide variation, and with metrics of health and overall reproductive fitness.}, } @article {pmid24012123, year = {2013}, author = {Lentz, CS and Stumpfe, D and Bajorath, J and Famulok, M and Hoerauf, A and Pfarr, KM}, title = {New chemotypes for wALADin1-like inhibitors of delta-aminolevulinic acid dehydratase from Wolbachia endobacteria.}, journal = {Bioorganic & medicinal chemistry letters}, volume = {23}, number = {20}, pages = {5558-5562}, doi = {10.1016/j.bmcl.2013.08.052}, pmid = {24012123}, issn = {1464-3405}, mesh = {Animals ; Benzimidazoles/chemical synthesis/*chemistry/metabolism ; Brugia malayi/enzymology ; Enzyme Inhibitors/chemical synthesis/*chemistry/metabolism ; Filaricides/chemical synthesis/*chemistry/metabolism ; Kinetics ; Porphobilinogen Synthase/*antagonists & inhibitors/metabolism ; Protein Binding ; Quinolines/chemistry ; Structure-Activity Relationship ; Thiophenes/chemical synthesis/*chemistry/metabolism ; Wolbachia/*enzymology ; }, abstract = {Substituted benzimidazoles of the wALADin1-family have recently been identified as a new class of species-selective inhibitors of delta-aminolevulinic acid dehydratase (ALAD) from Wolbachia endobacteria of parasitic filarial worms. Due to its Wolbachia-dependent antifilarial activity, wALADin1 is a starting point for the development of new drugs against filarial nematodes. We now present several other chemotypes of ALAD inhibitors that have been identified based upon their molecular similarity to wALADin1. A tricyclic quinoline derivative (wALADin2) with a different inhibitory mechanism and improved inhibitory potency and selectivity may represent an improved drug lead candidate.}, } @article {pmid24006922, year = {2013}, author = {Dumas, E and Atyame, CM and Milesi, P and Fonseca, DM and Shaikevich, EV and Unal, S and Makoundou, P and Weill, M and Duron, O}, title = {Population structure of Wolbachia and cytoplasmic introgression in a complex of mosquito species.}, journal = {BMC evolutionary biology}, volume = {13}, number = {}, pages = {181}, pmid = {24006922}, issn = {1471-2148}, mesh = {Animals ; Biological Evolution ; Culex/classification/*microbiology ; DNA, Mitochondrial/genetics ; Genetic Variation ; Haplotypes ; Polymerase Chain Reaction ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {BACKGROUND: The maternally inherited bacterium Wolbachia often acts as a subtle parasite that manipulates insect reproduction, resulting potentially in reproductive isolation between host populations. Whilst distinct Wolbachia strains are documented in a group of evolutionarily closely related mosquitoes known as the Culex pipiens complex, their impact on mosquito population genetics remains unclear. To this aim, we developed a PCR-RFLP test that discriminates the five known Wolbachia groups found in this host complex. We further examined the Wolbachia genetic diversity, the variability in the coinherited host mitochondria and their partitioning among members of the Cx. pipiens complex, in order to assess the impact of Wolbachia on host population structure.

RESULTS: There was a strong association between Wolbachia and mitochondrial haplotypes indicating a stable co-transmission in mosquito populations. Despite evidence that members of the Cx. pipiens complex are genetically distinct on the basis of nuclear DNA, the association of Wolbachia and mtDNA with members of the Cx. pipiens complex were limited. The Wolbachia wPip-I group, by far the most common, was associated with divergent Cx. pipiens members, including Cx. quinquefasciatus, Cx. pipiens pipiens form pipiens and Cx. pipiens pipiens form molestus. Four other wPip groups were also found in mosquito populations and all were shared between diverse Cx. pipiens members.

CONCLUSION: This data overall supports the hypothesis that wPip infections, and their allied mitochondria, are associated with regular transfers between Cx. pipiens members rather than specific host associations. Overall, this is suggestive of a recent and likely ongoing cytoplasmic introgression through hybridization events across the Cx. pipiens complex.}, } @article {pmid24002991, year = {2014}, author = {Lashkari, M and Manzari, S and Sahragard, A and Malagnini, V and Boykin, LM and Hosseini, R}, title = {Global genetic variation in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae) and the endosymbiont Wolbachia: links between Iran and the USA detected.}, journal = {Pest management science}, volume = {70}, number = {7}, pages = {1033-1040}, doi = {10.1002/ps.3643}, pmid = {24002991}, issn = {1526-4998}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Electron Transport Complex IV/genetics ; *Genetic Variation ; Hemiptera/*genetics/*microbiology ; Insect Proteins/genetics ; Iran ; Microsatellite Repeats ; Mitochondrial Proteins/genetics ; Molecular Sequence Data ; Pakistan ; Phylogeny ; Sequence Analysis, DNA ; United States ; Wolbachia/*genetics/*physiology ; }, abstract = {BACKGROUND: The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is one of the most serious pests of citrus in the world, because it transmits the pathogen that causes citrus greening disease. To determine genetic variation among geographic populations of D. citri, microsatellite markers, mitochondrial gene cytochrome oxidase I (mtCOI) and the Wolbachia-Diaphorina, wDi, gene wsp sequence data were used to characterize Iranian and Pakistani populations. Also, a Bayesian phylogenetic technique was utilized to elucidate the relationships among the sequences data in this study and all mtCOI and wsp sequence data available in GenBank and the Wolbachia database.

RESULTS: Microsatellite markers revealed significant genetic differentiation among Iranian populations, as well as between Iranian and Pakistani populations (FST  = 0.0428, p < 0.01). Within Iran, the Sistan-Baluchestan population is significantly different from the Hormozgan (Fareghan) and Fars populations. By contrast, mtCOI data revealed two polymorphic sites separating the sequences from Iran and Pakistan. Global phylogenetic analyses showed that D. citri populations in Iran, India, Saudi Arabia, Brazil, Mexico, Florida and Texas (USA) are similar. Wolbachia, wDi, wsp sequences were similar among Iranian populations, but different between Iranian and Pakistani populations.

CONCLUSION: The South West Asia (SWA) group is the most likely source of the introduced Iranian populations of D. citri. This assertion is also supported by the sequence similarity of the Wolbachia, wDi, strains from the Florida, USA and Iranian D. citri. These results should be considered when looking for biological controls in either country.}, } @article {pmid23986574, year = {2013}, author = {Rancès, E and Johnson, TK and Popovici, J and Iturbe-Ormaetxe, I and Zakir, T and Warr, CG and O'Neill, SL}, title = {The toll and Imd pathways are not required for wolbachia-mediated dengue virus interference.}, journal = {Journal of virology}, volume = {87}, number = {21}, pages = {11945-11949}, pmid = {23986574}, issn = {1098-5514}, mesh = {Animals ; Dengue Virus/*growth & development ; Drosophila Proteins/genetics/*immunology ; Drosophila melanogaster/immunology/*microbiology/*virology ; Female ; Immunity, Innate ; *Microbial Interactions ; Mutation ; Signal Transduction ; Toll-Like Receptors/genetics/*immunology ; Wolbachia/*growth & development ; }, abstract = {Wolbachia blocks dengue virus replication in Drosophila melanogaster as well as in Aedes aegypti. Using the Drosophila model and mutations in the Toll and Imd pathways, we showed that neither pathway is required for expression of the dengue virus-blocking phenotype in the Drosophila host. This provides additional evidence that the mechanistic basis of Wolbachia-mediated dengue virus blocking in insects is more complex than simple priming of the host insect innate immune system.}, } @article {pmid23975837, year = {2013}, author = {Sanada-Morimura, S and Matsumura, M and Noda, H}, title = {Male killing caused by a Spiroplasma symbiont in the small brown planthopper, Laodelphax striatellus.}, journal = {The Journal of heredity}, volume = {104}, number = {6}, pages = {821-829}, doi = {10.1093/jhered/est052}, pmid = {23975837}, issn = {1465-7333}, mesh = {Animals ; Female ; Genes, Insect ; Hemiptera/classification/drug effects/genetics/*microbiology ; Male ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sex Ratio ; *Spiroplasma/drug effects/physiology ; Symbiosis ; }, abstract = {Spiroplasma-mediated late male killing was found in the small brown planthopper, Laodelphax striatellus. Female-biased colonies (maternal lines, N = 4) were established from planthoppers collected in Taiwan and Japan. This sex ratio distortion was maternally inherited (sex ratio of total number of progenies [female:male]: 488:0 in F1, 198:7 in F2, 407:0 in F3; likelihood ratio test of all generations, P < 0.0001) and caused by male death during nymphal stages. The female-biased colonies were doubly infected with Spiroplasma and Wolbachia, and the non-biased colonies were infected solely with Wolbachia. Antibiotic treatment resulted in a normal sex ratio, strongly suggesting that bacteria are manipulating host reproduction. Spiroplasma-singly-infected planthopper colonies created by the antibiotic treatment produced progeny with strongly female-biased sex ratios (181:2; likelihood ratio test, χ(2) = 231.6, P < 0.0001). This is the first report of Spiroplasma-mediated male killing in hemimetabolous insects.}, } @article {pmid23967363, year = {2013}, author = {Medlock, J and Atkins, KE and Thomas, DN and Aksoy, S and Galvani, AP}, title = {Evaluating paratransgenesis as a potential control strategy for African trypanosomiasis.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {8}, pages = {e2374}, pmid = {23967363}, issn = {1935-2735}, support = {/WT_/Wellcome Trust/United Kingdom ; GM070694/GM/NIGMS NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; AI068932/AI/NIAID NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; U01 GM070694/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Communicable Disease Control/*methods ; Enterobacteriaceae/*growth & development ; Female ; Humans ; Male ; Microbial Interactions ; Models, Theoretical ; Trypanosomiasis, African/*epidemiology/*prevention & control/transmission ; Tsetse Flies/*microbiology/*parasitology ; Wolbachia/*growth & development ; }, abstract = {Genetic-modification strategies are currently being developed to reduce the transmission of vector-borne diseases, including African trypanosomiasis. For tsetse, the vector of African trypanosomiasis, a paratransgenic strategy is being considered: this approach involves modification of the commensal symbiotic bacteria Sodalis to express trypanosome-resistance-conferring products. Modified Sodalis can then be driven into the tsetse population by cytoplasmic incompatibility (CI) from Wolbachia bacteria. To evaluate the effectiveness of this paratransgenic strategy in controlling African trypanosomiasis, we developed a three-species mathematical model of trypanosomiasis transmission among tsetse, humans, and animal reservoir hosts. Using empirical estimates of CI parameters, we found that paratransgenic tsetse have the potential to eliminate trypanosomiasis, provided that any extra mortality caused by Wolbachia colonization is low, that the paratransgene is effective at protecting against trypanosome transmission, and that the target tsetse species comprises a large majority of the tsetse population in the release location.}, } @article {pmid23960254, year = {2013}, author = {Comandatore, F and Sassera, D and Montagna, M and Kumar, S and Koutsovoulos, G and Thomas, G and Repton, C and Babayan, SA and Gray, N and Cordaux, R and Darby, A and Makepeace, B and Blaxter, M}, title = {Phylogenomics and analysis of shared genes suggest a single transition to mutualism in Wolbachia of nematodes.}, journal = {Genome biology and evolution}, volume = {5}, number = {9}, pages = {1668-1674}, pmid = {23960254}, issn = {1759-6653}, support = {095831//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Bacterial Proteins/genetics ; Genome, Bacterial ; Genomics ; Host-Pathogen Interactions/*genetics ; Humans ; Nematoda/*microbiology ; *Phylogeny ; Symbiosis/genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia, endosymbiotic bacteria of the order Rickettsiales, are widespread in arthropods but also present in nematodes. In arthropods, A and B supergroup Wolbachia are generally associated with distortion of host reproduction. In filarial nematodes, including some human parasites, multiple lines of experimental evidence indicate that C and D supergroup Wolbachia are essential for the survival of the host, and here the symbiotic relationship is considered mutualistic. The origin of this mutualistic endosymbiosis is of interest for both basic and applied reasons: How does a parasite become a mutualist? Could intervention in the mutualism aid in treatment of human disease? Correct rooting and high-quality resolution of Wolbachia relationships are required to resolve this question. However, because of the large genetic distance between Wolbachia and the nearest outgroups, and the limited number of genomes so far available for large-scale analyses, current phylogenies do not provide robust answers. We therefore sequenced the genome of the D supergroup Wolbachia endosymbiont of Litomosoides sigmodontis, revisited the selection of loci for phylogenomic analyses, and performed a phylogenomic analysis including available complete genomes (from isolates in supergroups A, B, C, and D). Using 90 orthologous genes with reliable phylogenetic signals, we obtained a robust phylogenetic reconstruction, including a highly supported root to the Wolbachia phylogeny between a (A + B) clade and a (C + D) clade. Although we currently lack data from several Wolbachia supergroups, notably F, our analysis supports a model wherein the putatively mutualist endosymbiotic relationship between Wolbachia and nematodes originated from a single transition event.}, } @article {pmid23955974, year = {2013}, author = {Hassan, SS and Idris, E and Majerus, ME}, title = {Male-killer dynamics in the tropical butterfly, Acraea encedana (Lepidoptera: Nymphalidae).}, journal = {Insect science}, volume = {20}, number = {6}, pages = {717-722}, doi = {10.1111/j.1744-7917.2012.01570.x}, pmid = {23955974}, issn = {1744-7917}, mesh = {Animals ; Butterflies/*microbiology ; Female ; *Host-Pathogen Interactions ; Male ; Seasons ; *Sex Ratio ; Tropical Climate ; Uganda ; Wolbachia/*physiology ; }, abstract = {Sex ratio distortion in the tropical butterfly Acraea encedana is caused by infection with a male-killing bacterium of the genus Wolbachia. Previous research on this species has reported extreme female bias, high bacterial prevalences, and full sex role reversal. In this paper, we provide an assessment for the dynamics of the male-killer, based on a survey for sex ratios and Wolbachia prevalences among wild populations of A. encedana in Uganda. The study reveals that Wolbachia infection showed considerable variation over both spatial and temporal scales.}, } @article {pmid23955885, year = {2013}, author = {Zhang, KJ and Han, X and Hong, XY}, title = {Various infection status and molecular evidence for horizontal transmission and recombination of Wolbachia and Cardinium among rice planthoppers and related species.}, journal = {Insect science}, volume = {20}, number = {3}, pages = {329-344}, doi = {10.1111/j.1744-7917.2012.01537.x}, pmid = {23955885}, issn = {1744-7917}, mesh = {Animals ; DNA, Bacterial/genetics ; Gene Expression Regulation, Bacterial/physiology ; Hemiptera/*microbiology ; Nucleic Acid Amplification Techniques ; Oryza ; Recombination, Genetic ; Species Specificity ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia and Cardinium are widely distributed and are considered important for their ability to disturb reproduction and affect other fitness-related traits of their hosts. By using multilocus sequence typing (MLST), RFLP (restriction fragment length polymorphism) and 16S ribosomal DNA gene sequencing methods, we extensively surveyed Wolbachia and Cardinium infection status of four predominant rice planthoppers and one kind of leafhopper in different rice fields. The results demonstrated that Sogatella furcifera (Horváth) and Laodelphax striatellus (Fallén) were infected with the same Wolbachia strain (wStri), while Nilaparvata lugens (Stål) and its closely related species Nilaparvata muiri China were infected with two phylogeneticlly distant strains, wLug and wMui, respectively. Three new Wolbachia strains (provisionally named wMfas1, wMfas2 and wMfas3) were detected in the leafhopper Macrosteles fascifrons (Stål). Only S. furcifera was co-infected with Cardinium, which indicated that the distribution of Cardinium in these rice planthoppers was narrower than that of Wolbachia. Unambiguous intragenic recombination events among these Wolbachia strains and incongruent phylogenetic relationships show that the connections between different Wolbachia strains and hosts were more complex than we expected. These results suggest that horizontal transmission and host associated specialization are two factors affecting Wolbachia and Cardinium infections among planthoppers and their related species.}, } @article {pmid23955860, year = {2013}, author = {Bing, XL and Ruan, YM and Rao, Q and Wang, XW and Liu, SS}, title = {Diversity of secondary endosymbionts among different putative species of the whitefly Bemisia tabaci.}, journal = {Insect science}, volume = {20}, number = {2}, pages = {194-206}, doi = {10.1111/j.1744-7917.2012.01522.x}, pmid = {23955860}, issn = {1744-7917}, mesh = {Animals ; *Bacteria/genetics ; *Bacterial Physiological Phenomena ; *Biodiversity ; Evolution, Molecular ; Hemiptera/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; *Symbiosis ; }, abstract = {Endosymbionts are important components of arthropod biology. The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a cryptic species complex composed of ≥ 28 putative species. In addition to the primary endosymbiont Portiera aleyrodidarum, six secondary endosymbionts (S-endosymbionts), Hamiltonella, Rickettsia, Wolbachia, Cardinium, Arsenophonus and Fritschea, have been identified in B. tabaci thus far. Here, we tested five of the six S-endosymbiont lineages (excluding Fritschea) from 340 whitely individuals representing six putative species from China. Hamiltonella was detected only in the two exotic invaders, Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED). Rickettsia was absent in Asia II 1 and MED, scarce in Asia II 3 (13%), but abundant in Asia II 7 (63.2%), China 1 (84.7%) and MEAM1 (100%). Wolbachia, Cardinium and Arsenophonus were absent in the invasive MEAM1 and MED but mostly abundant in the native putative species. Furthermore, phylogenetic analyses revealed that some S-endosymbionts have several clades and different B. tabaci putative species can harbor different clades of a given S-endosymbiont, demonstrating further the complexity of S-endosymbionts in B. tabaci. All together, our results demonstrate the variation and diversity of S-endosymbionts in different putative species of B. tabaci, especially between invasive and native whiteflies.}, } @article {pmid23951381, year = {2013}, author = {Ye, YH and Woolfit, M and Rancès, E and O'Neill, SL and McGraw, EA}, title = {Wolbachia-associated bacterial protection in the mosquito Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {8}, pages = {e2362}, pmid = {23951381}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; *Immunity, Innate ; Microbial Interactions ; *Symbiosis ; Wolbachia/growth & development/immunology/*physiology ; }, abstract = {BACKGROUND: Wolbachia infections confer protection for their insect hosts against a range of pathogens including bacteria, viruses, nematodes and the malaria parasite. A single mechanism that might explain this broad-based pathogen protection is immune priming, in which the presence of the symbiont upregulates the basal immune response, preparing the insect to defend against subsequent pathogen infection. A study that compared natural Wolbachia infections in Drosophila melanogaster with the mosquito vector Aedes aegypti artificially transinfected with the same strains has suggested that innate immune priming may only occur in recent host-Wolbachia associations. This same study also revealed that while immune priming may play a role in viral protection it cannot explain the entirety of the effect.

METHODOLOGY/FINDINGS: Here we assess whether the level of innate immune priming induced by different Wolbachia strains in A. aegypti is correlated with the degree of protection conferred against bacterial pathogens. We show that Wolbachia strains wMel and wMelPop, currently being tested for field release for dengue biocontrol, differ in their protective abilities. The wMelPop strain provides stronger, more broad-based protection than wMel, and this is likely explained by both the higher induction of immune gene expression and the strain-specific activation of particular genes. We also show that Wolbachia densities themselves decline during pathogen infection, likely as a result of the immune induction.

CONCLUSIONS/SIGNIFICANCE: This work shows a correlation between innate immune priming and bacterial protection phenotypes. The ability of the Toll pathway, melanisation and antimicrobial peptides to enhance viral protection or to provide the basis of malaria protection should be further explored in the context of this two-strain comparison. This work raises the questions of whether Wolbachia may improve the ability of wild mosquitoes to survive pathogen infection or alter the natural composition of gut flora, and thus have broader consequences for host fitness.}, } @article {pmid23933013, year = {2013}, author = {Suh, E and Dobson, SL}, title = {Reduced competitiveness of Wolbachia infected Aedes aegypti larvae in intra- and inter-specific immature interactions.}, journal = {Journal of invertebrate pathology}, volume = {114}, number = {2}, pages = {173-177}, pmid = {23933013}, issn = {1096-0805}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*physiology ; Animals ; Host-Pathogen Interactions/*physiology ; Larva/microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacteria that frequently infect a diverse range of arthropod species. Empirical and theoretical studies examining Wolbachia invasiveness have emphasized Wolbachia effects on adult hosts, but recent studies show that Wolbachia impacts on immature hosts can be important also. Here, we have examined for effects of Wolbachia infection in Aedes aegypti. Specifically, differential survivorship is observed when young larvae (1st instar) are exposed to older Aedes albopictus larvae (4th instar) or con-specific larvae. In an additional experiment, we have examined for differential behavior and observed that Wolbachia-infected larvae differ from uninfected larvae in their reaction to light stimulation. Our results support a hypothesized effect of Wolbachia on A. aegypti larval behavior. The results are discussed in relation to the ability of Wolbachia to invade natural populations and recently applied public health strategies that target the replacement or suppression of this important disease vector.}, } @article {pmid23927410, year = {2013}, author = {Jäckel, R and Mora, D and Dobler, S}, title = {Evidence for selective sweeps by Wolbachia infections: phylogeny of Altica leaf beetles and their reproductive parasites.}, journal = {Molecular ecology}, volume = {22}, number = {16}, pages = {4241-4255}, doi = {10.1111/mec.12389}, pmid = {23927410}, issn = {1365-294X}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Coleoptera/genetics/*microbiology/physiology ; DNA, Mitochondrial/*genetics ; Evolution, Molecular ; Female ; Genetic Variation ; Haplotypes ; Host-Parasite Interactions/*genetics ; *Hybridization, Genetic ; Molecular Sequence Data ; *Phylogeny ; Plant Leaves/parasitology ; Reproduction/*genetics/physiology ; Selection, Genetic ; Sequence Analysis, DNA ; Sex Ratio ; Symbiosis ; Wolbachia/classification/genetics/*pathogenicity ; }, abstract = {Infections with maternally inherited Wolbachia bacteria may have dramatic influences on reproductive traits and speciation patterns of their hosts. We here show that in the beetle genus Altica, infection has influenced phylogenetic patterns of the host's mtDNA and different strains led to repeated selective sweeps. By comparing a COI/II-based phylogeny of the hosts with a phylogeny of the bacteria based on ftsZ, we show that cospeciation is rare and restricted to few recently diverged species. While in general each species apparently harbours a single Wolbachia strain, Altica lythri presents a strikingly different pattern: in the polyphyletic species, three highly divergent mtDNA haplotypes (2.1-4.6% p-distance) are coupled with three different Wolbachia strains (wLytA1, wLytA2 and wLytB). These haplotypes and Wolbachia strains are widely distributed and mostly found in sympatry. A phylogeny based on microsatellite data supports the monophyly of A. lythri. The discrepancy between mtDNA and nuclear phylogeny may best be explained by interspecific hybridization that led to introgression of mtDNA coupled with a different Wolbachia strain. Selective sweeps apparently drove the introgressed haplotypes to widespread distribution. As for effects of Wolbachia on reproduction, infection with wLytA1 appears to be correlated with a substantial sex ratio distortion, which was most prominent in A. lythri.}, } @article {pmid23924682, year = {2013}, author = {Wamwiri, FN and Alam, U and Thande, PC and Aksoy, E and Ngure, RM and Aksoy, S and Ouma, JO and Murilla, GA}, title = {Wolbachia, Sodalis and trypanosome co-infections in natural populations of Glossina austeni and Glossina pallidipes.}, journal = {Parasites & vectors}, volume = {6}, number = {1}, pages = {232}, pmid = {23924682}, issn = {1756-3305}, support = {R03TW008413/TW/FIC NIH HHS/United States ; D43 TW007391/TW/FIC NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; R03 TW008413/TW/FIC NIH HHS/United States ; D43TW007391/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; Coinfection/microbiology/parasitology/veterinary ; Enterobacteriaceae/genetics/isolation & purification/*physiology ; Female ; Insect Vectors/microbiology/parasitology/physiology ; Male ; Symbiosis ; Trypanosoma/genetics/isolation & purification/*physiology ; Tsetse Flies/*microbiology/*parasitology/physiology ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {BACKGROUND: Tsetse flies harbor at least three bacterial symbionts: Wigglesworthia glossinidia, Wolbachia pipientis and Sodalis glossinidius. Wigglesworthia and Sodalis reside in the gut in close association with trypanosomes and may influence establishment and development of midgut parasite infections. Wolbachia has been shown to induce reproductive effects in infected tsetse. This study was conducted to determine the prevalence of these endosymbionts in natural populations of G. austeni and G. pallidipes and to assess the degree of concurrent infections with trypanosomes.

METHODS: Fly samples analyzed originated from Kenyan coastal forests (trapped in 2009-2011) and South African G. austeni collected in 2008. The age structure was estimated by standard methods. G. austeni (n=298) and G. pallidipes (n= 302) were analyzed for infection with Wolbachia and Sodalis using PCR. Trypanosome infection was determined either by microscopic examination of dissected organs or by PCR amplification.

RESULTS: Overall we observed that G. pallidipes females had a longer lifespan (70 d) than G. austeni (54 d) in natural populations. Wolbachia infections were present in all G. austeni flies analysed, while in contrast, this symbiont was absent from G. pallidipes. The density of Wolbachia infections in the Kenyan G. austeni population was higher than that observed in South African flies. The infection prevalence of Sodalis ranged from 3.7% in G. austeni to about 16% in G. pallidipes. Microscopic examination of midguts revealed an overall trypanosome infection prevalence of 6% (n = 235) and 5% (n = 552), while evaluation with ITS1 primers indicated a prevalence of about 13% (n = 296) and 10% (n = 302) in G. austeni and G. pallidipes, respectively. The majority of infections (46%) were with T. congolense. Co-infection with all three organisms was observed at 1% and 3.3% in G. austeni and G. pallidipes, respectively. Eleven out of the thirteen (85%) co-infected flies harboured T. congolense and T. simiae parasites. While the association between trypanosomes and Sodalis infection was statistically significant in G. pallidipes (P = 0.0127), the number of co-infected flies was too few for a definite conclusion.

CONCLUSIONS: The tsetse populations analyzed differed in the prevalence of symbionts, despite being sympatric and therefore exposed to identical environmental factors. The density of infections with Wolbachia also differed between G. austeni populations. There were too few natural co-infections detected with the Sodalis and trypanosomes to suggest extensive inter-relations between these infections in natural populations. We discuss these findings in the context of potential symbiont-mediated control interventions to reduce parasite infections and/or fly populations.}, } @article {pmid23921743, year = {2013}, author = {}, title = {Modulating malaria with Wolbachia.}, journal = {Nature medicine}, volume = {19}, number = {8}, pages = {974-975}, pmid = {23921743}, issn = {1546-170X}, mesh = {Animals ; Anopheles/microbiology ; Disease Vectors ; Humans ; Malaria/*prevention & control/transmission ; Plasmodium falciparum/physiology ; Virus Replication ; Wolbachia/*physiology ; }, } @article {pmid23909372, year = {2013}, author = {Martin, OY and Puniamoorthy, N and Gubler, A and Wimmer, C and Germann, C and Bernasconi, MV}, title = {Infections with the microbe Cardinium in the Dolichopodidae and other Empidoidea.}, journal = {Journal of insect science (Online)}, volume = {13}, number = {}, pages = {47}, pmid = {23909372}, issn = {1536-2442}, mesh = {Animals ; Bacteroidetes/*isolation & purification ; Diptera/*microbiology ; Symbiosis ; }, abstract = {Maternally transmitted reproductive parasites such as Wolbachia and Cardinium can drastically reshape reproduction in their hosts. Beyond skewing sex ratios towards females, these microbes can also cause cytoplasmic incompatibility. Wolbachia probably infects two thirds of insects, but far less is known about the occurrence or action of other bacteria with potentially similar effects. In contrast with the two more widespread reproductive parasites, Wolbachia and Spiroplasma, far less is known of infections with Cardinium (Bacteroidetes) and possible consequences in the Diptera. Here, in an extensive survey, 244 dipteran species from 67 genera belonging to the Dolichopodidae, Empididae, and Hybotidae were assessed for the presence of the microbe Cardinium. Although 130 of the species screened tested positive (ca. 53%), the presence of Cardinium could only be confirmed in 10 species (ca. 4%) based on analysis of sequences. Numerous additional sequences were found to be assignable to known or unknown Bacteroidetes. Considering the known issues concerning specificity of Cardinium primers and the phylogenetic uncertainties surrounding this microbe, the actual prevalence of this symbiont is worthy of further scrutiny. Potential directions for future research on Cardinium-host interactions in Diptera and in general are discussed.}, } @article {pmid23906981, year = {2013}, author = {Müller, MJ and Dörr, NC and Deprá, M and Schmitz, HJ and Valiati, VH and Valente, VL}, title = {Reevaluating the infection status by the Wolbachia endosymbiont in Drosophila Neotropical species from the willistoni subgroup.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {19}, number = {}, pages = {232-239}, doi = {10.1016/j.meegid.2013.07.022}, pmid = {23906981}, issn = {1567-7257}, mesh = {Animals ; DNA, Bacterial/analysis/genetics ; Drosophila/*genetics/*microbiology ; Female ; Genes, Insect/genetics ; Male ; Minisatellite Repeats/genetics ; Symbiosis/*genetics ; Wolbachia/classification/*genetics/isolation & purification ; }, abstract = {Infections by the endosymbiotic bacterium Wolbachia developed a rapid global expansion within Old World Drosophila species, ultimately infecting also Neotropical species. In this sense, screenings are necessary to characterize new variants of Wolbachia or new hosts, and also in order to map the dynamics of already known infections. In this paper, we performed a double screening approach that combined Dot-blot and PCR techniques in order to reevaluate the infection status by Wolbachia in species from the willistoni subgroup of Drosophila. Genomic DNA from isofemale lines descendent from females collected in the Amazonian Rainforest (n=91) were submitted to Dot-blot, and were positive for Wolbachia, producing a gradient of hybridization signals, suggesting different infection levels, which was further confirmed through quantitative PCR. Samples with a strong signal in the Dot-blot easily amplified in the wsp-PCR, unlike most of the samples with a medium to weak signal. It was possible to molecularly characterize three Drosophila equinoxialis isofemale lines that were found to be infected in a low density by a wMel-like Wolbachia strain, which was also verified in a laboratory line of Drosophila paulistorum Amazonian. We also found Drosophila tropicalis to be infected with the wAu strain and a Drosophila paulistorum Andean-Brazilian semispecies laboratory line to be infected with a wAu-like Wolbachia. Moreover, we observed that all Drosophila willistoni samples tested with the VNTR-141 marker harbor the same Wolbachia variant, wWil, either in populations from the South or the North of Brazil. Horizontal transfer events involving species of Old World immigrants and Neotropical species of the willistoni subgroup are discussed.}, } @article {pmid23905733, year = {2013}, author = {Singh, ST and Kumar, J and Thomas, A and Ramamurthy, VV and Rajagopal, R}, title = {Detection and localization of Rickettsia sp in mealybug.}, journal = {Environmental entomology}, volume = {42}, number = {4}, pages = {711-716}, doi = {10.1603/EN13032}, pmid = {23905733}, issn = {1938-2936}, mesh = {Animals ; DNA, Bacterial/genetics/metabolism ; Female ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; India ; Introduced Species ; Microbiota ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics/metabolism ; Rickettsia/*genetics/*physiology ; Sequence Homology ; Symbiosis ; }, abstract = {Mealybug, Phenacoccus solenopsis Tinsley, is a sap-sucking hemipteran insect. It is an agricultural pest that is now widely distributed in India. In this study we report the presence of Rickettsia from P. solenopsis. We constructed a 16S rRNA gene library to study the bacterial diversity associated with this insect and we found that all the clones from the library were only of Candidatus Tremblaya phenacola. This study also highlights that the normal protocol adopted to study the bacterial diversity from environmental sample, by preparation of a 16S rRNA gene library, does not work when the bacterial population is highly skewed in favor of one bacteria (primary endosymbiont in this case). Hence, we used bacterial genus specific polymerase chain reaction primers to test the presence of any of the widely known secondary endosymbionts associated with insects. We tested for the presence of Cardinium, Rickettsia, Wolbachia, and Arsenophonus in P. solenopsis collected from 10 different locations across India. Only Rickettsia was detected from four locations while we were not able to find any other bacteria. We confirmed the presence of these bacteria by localizing Rickettsia and the primary endosmbiont, Candidatus Tremblaya sp. to the bacteriocyte of P. solenopsis using fluorescent in situ hybridization.}, } @article {pmid23879258, year = {2013}, author = {Reumer, BM and van Alphen, JJ and Kraaijeveld, K}, title = {Population genetics of Wolbachia-infected, parthenogenetic and uninfected, sexual populations of Tetrastichus coeruleus (Hymenoptera: Eulophidae).}, journal = {Molecular ecology}, volume = {22}, number = {17}, pages = {4433-4444}, doi = {10.1111/mec.12397}, pmid = {23879258}, issn = {1365-294X}, mesh = {Animals ; Belgium ; Cell Nucleus/genetics ; Cluster Analysis ; DNA, Mitochondrial/genetics ; Female ; Genes, Insect ; *Genetic Variation ; *Genetics, Population ; Haplotypes ; Male ; Microsatellite Repeats ; Models, Genetic ; Molecular Sequence Data ; Netherlands ; Parthenogenesis ; Symbiosis ; Wasps/*genetics/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria known to manipulate the reproduction of their hosts. These manipulations are expected to have consequences on the population genetics of the host, such as heterozygosity levels, genetic diversity and gene flow. The parasitoid wasp Tetrastichus coeruleus has populations that are infected with parthenogenesis-inducing Wolbachia and populations that are not infected. We studied the population genetics of T. coeruleus between and within Wolbachia-infected and uninfected populations, using nuclear microsatellites and mitochondrial DNA. We expected reduced genetic diversity in both DNA types in infected populations. However, migration and gene flow could introduce new DNA variants into populations. We therefore paid special attention to individuals with unexpected (genetic) characteristics. Based on nuclear and mitochondrial DNA, two genetic clusters were evident: a thelytokous cluster containing all Wolbachia-infected, parthenogenetic populations and an arrhenotokous cluster containing all uninfected, sexual populations. Nuclear and mitochondrial DNA did not exhibit concordant patterns of variation, although there was reduced genetic diversity in infected populations for both DNA types. Within the thelytokous cluster, there was nuclear DNA variation, but no mitochondrial DNA variation. This nuclear DNA variation may be explained by occasional sex between infected females and males, by horizontal transmission of Wolbachia, and/or by novel mutations. Several females from thelytokous populations were uninfected and/or heterozygous for microsatellite loci. These unexpected characteristics may be explained by migration, by inefficient transmission of Wolbachia, by horizontal transmission of Wolbachia, and/or by novel mutations. However, migration has not prevented the build-up of considerable genetic differentiation between thelytokous and arrhenotokous populations.}, } @article {pmid23866958, year = {2014}, author = {Taylor, MJ and Hoerauf, A and Townson, S and Slatko, BE and Ward, SA}, title = {Anti-Wolbachia drug discovery and development: safe macrofilaricides for onchocerciasis and lymphatic filariasis.}, journal = {Parasitology}, volume = {141}, number = {1}, pages = {119-127}, pmid = {23866958}, issn = {1469-8161}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/drug effects/microbiology/physiology ; Child ; Doxycycline/pharmacology ; Drug Discovery ; Elephantiasis, Filarial/*drug therapy/parasitology ; Female ; Filaricides/*pharmacology ; Humans ; Ivermectin/pharmacology ; Loiasis/*drug therapy/parasitology ; Onchocerca volvulus/drug effects/microbiology/physiology ; Onchocerciasis/*drug therapy/parasitology ; Pregnancy ; Symbiosis/drug effects ; Wolbachia/*drug effects/growth & development ; }, abstract = {Anti-Wolbachia therapy delivers safe macrofilaricidal activity with superior therapeutic outcomes compared to all standard anti-filarial treatments, with the added benefit of substantial improvements in clinical pathology. These outcomes can be achieved, in principle, with existing registered drugs, e.g. doxycycline, that are affordable, available to endemic communities and have well known, albeit population-limiting, safety profiles. The key barriers to using doxycycline as an mass drug administration (MDA) strategy for widespread community-based control are the logistics of a relatively lengthy course of treatment (4-6 weeks) and contraindications in children under eight years and pregnancy. Therefore, the primary goal of the anti-Wolbachia (A·WOL) consortium is to find drugs and regimens that reduce the period of treatment from weeks to days (7 days or less), and to find drugs which would be safe in excluded target populations (pregnancy and children). A secondary goal is to refine regimens of existing antibiotics suitable for a more restricted use, prior to the availability of a regimen that is compatible with MDA usage. For example, for use in the event of the emergence of drug-resistance, in individuals with high loiasis co-infection and at risk of severe adverse events (SAE) to ivermectin, or in post-MDA 'endgame scenarios', where test and treat strategies become more cost effective and deliverable.}, } @article {pmid23856508, year = {2013}, author = {Beckmann, JF and Fallon, AM}, title = {Detection of the Wolbachia protein WPIP0282 in mosquito spermathecae: implications for cytoplasmic incompatibility.}, journal = {Insect biochemistry and molecular biology}, volume = {43}, number = {9}, pages = {867-878}, pmid = {23856508}, issn = {1879-0240}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/genetics/*metabolism ; Culex/chemistry/*microbiology/*physiology ; Cytoplasm/*microbiology ; Female ; Infertility ; Male ; Molecular Sequence Data ; Ovary/microbiology ; Sequence Alignment ; Spermatozoa/chemistry/*microbiology ; Wolbachia/genetics/*metabolism ; }, abstract = {Cytoplasmic incompatibility (CI) is a conditional sterility induced by the bacterium Wolbachia pipientis that infects reproductive tissues in many arthropods. Although CI provides a potential tool to control insect vectors of arthropod-borne diseases, the molecular basis for CI induction is unknown. We hypothesized that a Wolbachia-encoded, CI-inducing factor would be enriched in sperm recovered from spermathecae of female mosquitoes. Using SDS-PAGE and mass spectrometry, we detected peptides from the 56 kDa hypothetical protein, encoded by wPip_0282, associated with sperm transferred to females by Wolbachia infected males. We also detected peptides from the same protein in Wolbachia infected ovaries. Homologs of wPip_0282 and the co-transcribed downstream gene, wPip_0283, occur as multiple divergent copies in genomes of CI-inducing strains of Wolbachia. The operon is located in a genomic context that includes mobile genetic elements. The absence of wPip_0282 and wPip_0283 homologs from genomes of Wolbachia in filarial nematodes, as well as other members of the Rickettsiales, suggests a role as a candidate CI effector.}, } @article {pmid23848578, year = {2013}, author = {Françoso, E and Arias, MC}, title = {Cytochrome c oxidase I primers for corbiculate bees: DNA barcode and mini-barcode.}, journal = {Molecular ecology resources}, volume = {13}, number = {5}, pages = {844-850}, doi = {10.1111/1755-0998.12135}, pmid = {23848578}, issn = {1755-0998}, mesh = {Animals ; Bees/*classification/*genetics ; DNA Barcoding, Taxonomic/*methods ; DNA Primers/genetics ; DNA, Mitochondrial/chemistry/*genetics ; Electron Transport Complex IV/*genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; }, abstract = {Bees (Apidae), of which there are more than 19 900 species, are extremely important for ecosystem services and economic purposes, so taxon identity is a major concern. The goal of this study was to optimize the DNA barcode technique based on the Cytochrome c oxidase (COI) mitochondrial gene region. This approach has previously been shown to be useful in resolving taxonomic inconsistencies and for species identification when morphological data are poor. Specifically, we designed and tested new primers and standardized PCR conditions to amplify the barcode region for bees, focusing on the corbiculate Apids. In addition, primers were designed to amplify small COI amplicons and tested with pinned specimens. Short barcode sequences were easily obtained for some Bombus century-old museum specimens and shown to be useful as mini-barcodes. The new primers and PCR conditions established in this study proved to be successful for the amplification of the barcode region for all species tested, regardless of the conditions of tissue preservation. We saw no evidence of Wolbachia or numts amplification by these primers, and so we suggest that these new primers are of broad value for corbiculate bee identification through DNA barcode.}, } @article {pmid23845310, year = {2013}, author = {LePage, D and Bordenstein, SR}, title = {Wolbachia: Can we save lives with a great pandemic?.}, journal = {Trends in parasitology}, volume = {29}, number = {8}, pages = {385-393}, pmid = {23845310}, issn = {1471-5007}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; T32 GM008554/GM/NIGMS NIH HHS/United States ; 5T32GM008554-1/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Culicidae/*microbiology ; Female ; Filariasis/*prevention & control ; Global Health ; Humans ; Insect Vectors/*microbiology ; Male ; Nematoda/*microbiology ; Pandemics ; Pest Control, Biological/*methods ; Reproduction ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia pipientis is the most common bacterial infection in the animal world and wields a vast influence on invertebrate reproduction, sex determination, speciation, and behavior worldwide. These avenues of research have made seminal gains, including the latest use of Wolbachia to alter mosquito populations and a strengthened focus on using anti-Wolbachia therapies against filarial nematode infections. This work is further bolstered by a more refined knowledge of Wolbachia biology spanning mechanisms to relevance. Here we tally the most up-to-date knowledge in the field and review the immense implications that this global infection has for the basic and applied life sciences.}, } @article {pmid23844658, year = {2013}, author = {Schuler, H and Bertheau, C and Egan, SP and Feder, JL and Riegler, M and Schlick-Steiner, BC and Steiner, FM and Johannesen, J and Kern, P and Tuba, K and Lakatos, F and Köppler, K and Arthofer, W and Stauffer, C}, title = {Evidence for a recent horizontal transmission and spatial spread of Wolbachia from endemic Rhagoletis cerasi (Diptera: Tephritidae) to invasive Rhagoletis cingulata in Europe.}, journal = {Molecular ecology}, volume = {22}, number = {15}, pages = {4101-4111}, doi = {10.1111/mec.12362}, pmid = {23844658}, issn = {1365-294X}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Disease Transmission, Infectious ; Electron Transport Complex IV/*genetics ; Genetic Variation ; Genotype ; Multilocus Sequence Typing ; Tephritidae/*microbiology ; Wolbachia/classification/*genetics ; }, abstract = {The widespread occurrence of Wolbachia in arthropods and nematodes suggests that this intracellular, maternally inherited endosymbiont has the ability to cross species boundaries. However, direct evidence for such a horizontal transmission of Wolbachia in nature is scarce. Here, we compare the well-characterized Wolbachia infection of the European cherry fruit fly, Rhagoletis cerasi, with that of the North American eastern cherry fruit fly, Rhagoletis cingulata, recently introduced to Europe. Molecular genetic analysis of Wolbachia based on multilocus sequence typing and the Wolbachia surface protein wsp showed that all R. cingulata individuals are infected with wCin2 identical to wCer2 in R. cerasi. In contrast, wCin1, a strain identical to wCer1 in R. cerasi, was present in several European populations of R. cingulata, but not in any individual from the United States. Surveys of R. cingulata from Germany and Hungary indicated that in some populations, the frequency of wCin1 increased significantly in just a few years with at least two independent horizontal transmission events. This is corroborated by the analysis of the mitochondrial cytochrome oxidase II gene that showed association of wCin1 with two distinct haplotypes in Germany, one of which is also infected with wCin1 in Hungary. In summary, our study provides strong evidence for a very recent inter-specific Wolbachia transmission with a subsequent spatial spread in field populations.}, } @article {pmid23840485, year = {2013}, author = {Ye, YH and Woolfit, M and Huttley, GA and Rancès, E and Caragata, EP and Popovici, J and O'Neill, SL and McGraw, EA}, title = {Infection with a Virulent Strain of Wolbachia Disrupts Genome Wide-Patterns of Cytosine Methylation in the Mosquito Aedes aegypti.}, journal = {PloS one}, volume = {8}, number = {6}, pages = {e66482}, pmid = {23840485}, issn = {1932-6203}, mesh = {Aedes/genetics/*microbiology ; Animals ; Cytosine/*metabolism ; *DNA Methylation ; Gene Expression ; Genes, Insect ; *Genome ; *Virulence ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: Cytosine methylation is one of several reversible epigenetic modifications of DNA that allow a greater flexibility in the relationship between genotype and phenotype. Methylation in the simplest models dampens gene expression by modifying regions of DNA critical for transcription factor binding. The capacity to methylate DNA is variable in the insects due to diverse histories of gene loss and duplication of DNA methylases. Mosquitoes like Drosophila melanogaster possess only a single methylase, DNMT2.

DESCRIPTION: Here we characterise the methylome of the mosquito Aedes aegypti and examine its relationship to transcription and test the effects of infection with a virulent strain of the endosymbiont Wolbachia on the stability of methylation patterns.

CONCLUSION: We see that methylation in the A. aegypti genome is associated with reduced transcription and is most common in the promoters of genes relating to regulation of transcription and metabolism. Similar gene classes are also methylated in aphids and honeybees, suggesting either conservation or convergence of methylation patterns. In addition to this evidence of evolutionary stability, we also show that infection with the virulent wMelPop Wolbachia strain induces additional methylation and demethylation events in the genome. While most of these changes seem random with respect to gene function and have no detected effect on transcription, there does appear to be enrichment of genes associated with membrane function. Given that Wolbachia lives within a membrane-bound vacuole of host origin and retains a large number of genes for transporting host amino acids, inorganic ions and ATP despite a severely reduced genome, these changes might represent an evolved strategy for manipulating the host environments for its own gain. Testing for a direct link between these methylation changes and expression, however, will require study across a broader range of developmental stages and tissues with methods that detect splice variants.}, } @article {pmid23836873, year = {2013}, author = {Snyder, AK and Rio, RV}, title = {Interwoven biology of the tsetse holobiont.}, journal = {Journal of bacteriology}, volume = {195}, number = {19}, pages = {4322-4330}, pmid = {23836873}, issn = {1098-5530}, mesh = {Animals ; Bacteria/*classification/genetics ; Biological Evolution ; Symbiosis ; Tsetse Flies/genetics/*microbiology ; }, abstract = {Microbial symbionts can be instrumental to the evolutionary success of their hosts. Here, we discuss medically significant tsetse flies (Diptera: Glossinidae), a group comprised of over 30 species, and their use as a valuable model system to study the evolution of the holobiont (i.e., the host and associated microbes). We first describe the tsetse microbiota, which, despite its simplicity, harbors a diverse range of associations. The maternally transmitted microbes consistently include two Gammaproteobacteria, the obligate mutualists Wigglesworthia spp. and the commensal Sodalis glossinidius, along with the parasitic Alphaproteobacteria Wolbachia. These associations differ in their establishment times, making them unique and distinct from previously characterized symbioses, where multiple microbial partners have associated with their host for a significant portion of its evolution. We then expand into discussing the functional roles and intracommunity dynamics within this holobiont, which enhances our understanding of tsetse biology to encompass the vital functions and interactions of the microbial community. Potential disturbances influencing the tsetse microbiome, including salivary gland hypertrophy virus and trypanosome infections, are highlighted. While previous studies have described evolutionary consequences of host association for symbionts, the initial steps facilitating their incorporation into a holobiont and integration of partner biology have only begun to be explored. Research on the tsetse holobiont will contribute to the understanding of how microbial metabolic integration and interdependency initially may develop within hosts, elucidating mechanisms driving adaptations leading to cooperation and coresidence within the microbial community. Lastly, increased knowledge of the tsetse holobiont may also contribute to generating novel African trypanosomiasis disease control strategies.}, } @article {pmid23828002, year = {2013}, author = {Parvizi, P and Bordbar, A and Najafzadeh, N}, title = {Detection of Wolbachia pipientis, including a new strain containing the wsp gene, in two sister species of Paraphlebotomus sandflies, potential vectors of zoonotic cutaneous leishmaniasis.}, journal = {Memorias do Instituto Oswaldo Cruz}, volume = {108}, number = {4}, pages = {414-420}, pmid = {23828002}, issn = {1678-8060}, mesh = {Animals ; Base Sequence ; Insect Vectors/*microbiology ; Iran ; Leishmaniasis, Cutaneous/transmission ; Molecular Sequence Data ; Phlebotomus/*microbiology ; Polymerase Chain Reaction ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Individual, naturally occurring Phlebotomus mongolensis and Phlebotomus caucasicus from Iran were screened for infections with the maternally inherited intracellular Rickettsia-like bacterium Wolbachia pipientis via targeting a major surface protein gene (wsp). The main objective of this study was to determine if W. pipientis could be detected in these species. The sandflies were screened using polymerase chain reaction to amplify a fragment of the Wolbachia surface protein gene. The obtained sequences were edited and aligned with database sequences to identify W. pipientis haplotypes. Two strains of Wolbachia were found. Strain Turk 54 (accession EU780683) is widespread and has previously been reported in Phlebotomus papatasi and other insects. Strain Turk 07 (accession KC576916) is a novel strain, found for first time in the two sister species. A-group strains of W. pipientis occur throughout much of the habitat of these sandflies. It is possible that Wolbachia is transferred via horizontal transmission. Horizontal transfer could shed light on sandfly control because Wolbachia is believed to drive a deleterious gene into sandflies that reduces their natural population density. With regard to our findings in this study, we can conclude that one species of sandfly can be infected with different Wolbachia strains and that different species of sandflies can be infected with a common strain.}, } @article {pmid23825950, year = {2013}, author = {Caragata, EP and Rancès, E and Hedges, LM and Gofton, AW and Johnson, KN and O'Neill, SL and McGraw, EA}, title = {Dietary cholesterol modulates pathogen blocking by Wolbachia.}, journal = {PLoS pathogens}, volume = {9}, number = {6}, pages = {e1003459}, pmid = {23825950}, issn = {1553-7374}, mesh = {*Aedes/metabolism/microbiology/virology ; Animals ; Cholesterol/metabolism/*pharmacology ; Dicistroviridae/*metabolism ; Dietary Fats/metabolism/*pharmacology ; Drosophila melanogaster ; Host-Pathogen Interactions/*drug effects/physiology ; Wolbachia/*physiology ; }, abstract = {The bacterial endosymbiont Wolbachia pipientis protects its hosts from a range of pathogens by limiting their ability to form infections inside the insect. This "pathogen blocking" could be explained by innate immune priming by the symbiont, competition for host-derived resources between pathogens and Wolbachia, or the direct modification of the cell or cellular environment by Wolbachia. Recent comparative work in Drosophila and the mosquito Aedes aegypti has shown that an immune response is not required for pathogen blocking, implying that there must be an additional component to the mechanism. Here we have examined the involvement of cholesterol in pathogen blocking using a system of dietary manipulation in Drosophila melanogaster in combination with challenge by Drosophila C virus (DCV), a common fly pathogen. We observed that flies reared on cholesterol-enriched diets infected with the Wolbachia strains wMelPop and wMelCS exhibited reduced pathogen blocking, with viral-induced mortality occurring 2-5 days earlier than flies reared on Standard diet. This shift toward greater virulence in the presence of cholesterol also corresponded to higher viral copy numbers in the host. Interestingly, an increase in dietary cholesterol did not have an effect on Wolbachia density except in one case, but this did not directly affect the strength of pathogen blocking. Our results indicate that host cholesterol levels are involved with the ability of Wolbachia-infected flies to resist DCV infections, suggesting that cholesterol contributes to the underlying mechanism of pathogen blocking.}, } @article {pmid23823081, year = {2013}, author = {Zhao, DX and Zhang, XF and Chen, DS and Zhang, YK and Hong, XY}, title = {Wolbachia-Host Interactions: Host Mating Patterns Affect Wolbachia Density Dynamics.}, journal = {PloS one}, volume = {8}, number = {6}, pages = {e66373}, pmid = {23823081}, issn = {1932-6203}, mesh = {Animals ; *Colony Count, Microbial ; Female ; Genes, Bacterial ; *Host-Pathogen Interactions ; In Situ Hybridization, Fluorescence ; Male ; Multilocus Sequence Typing ; Polymerase Chain Reaction ; *Sexual Behavior, Animal ; Tetranychidae/*microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacteria that infect a wide range of arthropods and cause an array of effects on host reproduction, fitness and mating behavior. Although our understanding of the Wolbachia-associated effects on hosts is rapidly expanding, our knowledge of the host factors that mediate Wolbachia dynamics is rudimentary. Here, we explore the interactions between Wolbachia and its host, the two-spotted spider mite Tetranychus urticae Koch. Our results indicate that Wolbachia induces strong cytoplasmic incompatibility (CI), increases host fecundity, but has no effects on the longevity of females and the mating competitiveness of males in T. urticae. Most importantly, host mating pattern was found to affect Wolbachia density dynamics during host aging. Mating of an uninfected mite of either sex with an infected mite attenuates the Wolbachia density in the infected mite. According to the results of Wolbachia localization, this finding may be associated with the tropism of Wolbachia for the reproductive tissue in adult spider mites. Our findings describe a new interaction between Wolbachia and their hosts.}, } @article {pmid23816508, year = {2013}, author = {Alphey, L and McKemey, A and Nimmo, D and Neira Oviedo, M and Lacroix, R and Matzen, K and Beech, C}, title = {Genetic control of Aedes mosquitoes.}, journal = {Pathogens and global health}, volume = {107}, number = {4}, pages = {170-179}, pmid = {23816508}, issn = {2047-7732}, mesh = {Aedes/genetics/microbiology/*physiology ; Animals ; *Insect Vectors ; Mosquito Control/*methods ; Population Density ; Wolbachia/growth & development ; }, abstract = {Aedes mosquitoes include important vector species such as Aedes aegypti, the major vector of dengue. Genetic control methods are being developed for several of these species, stimulated by an urgent need owing to the poor effectiveness of current methods combined with an increase in chemical pesticide resistance. In this review we discuss the various genetic strategies that have been proposed, their present status, and future prospects. We focus particularly on those methods that are already being tested in the field, including RIDL and Wolbachia-based approaches.}, } @article {pmid23816506, year = {2013}, author = {Benedict, MQ}, title = {Male mosquitoes make waves in paradise.}, journal = {Pathogens and global health}, volume = {107}, number = {4}, pages = {161}, pmid = {23816506}, issn = {2047-7732}, mesh = {Aedes/*microbiology ; Animals ; Female ; Male ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; Wolbachia/*pathogenicity ; }, } @article {pmid23807431, year = {2013}, author = {Body, M and Kaiser, W and Dubreuil, G and Casas, J and Giron, D}, title = {Leaf-miners co-opt microorganisms to enhance their nutritional environment.}, journal = {Journal of chemical ecology}, volume = {39}, number = {7}, pages = {969-977}, pmid = {23807431}, issn = {1573-1561}, mesh = {Animals ; *Carbohydrate Metabolism ; Cytokinins/*metabolism ; Female ; Homeostasis ; Larva/microbiology ; Male ; Malus/*metabolism ; Moths/*microbiology ; Plant Leaves/*metabolism ; Symbiosis ; Wolbachia ; }, abstract = {Organisms make the best of their mother's oviposition choices and utilize specific feeding options that meet energetic requirements and cope with environmental constraints. This is particularly true for leaf-miner insects that develop enclosed in the two epidermis layers of a single leaf for their entire larval life. Cytokinins (CKs) play a central role in plant physiology - including regulation of senescence and nutrient translocation - and, as such, can be the specific target of plant exploiters that manipulate plant primary metabolism. 'Green-islands' are striking examples of a CK-induced phenotype where green areas are induced by plant pathogens/insects in otherwise yellow senescent leaves. Here, we document how the leaf-miner caterpillar Phyllonorycter blancardella, working through an endosymbiotic bacteria, modifies phytohormonal profiles, not only on senescing (photosynthetically inactive) but also on normal (photosynthetically active) leaf tissues of its host plant (Malus domestica). This leaf physiological manipulation allows the insect to maintain sugar-rich green tissues and to create an enhanced nutritional microenvironment in an otherwise degenerating context. It also allows them to maintain a nutritional homeostasis even under distinct leaf environments. Our study also highlights that only larvae harboring bacterial symbionts contain significant amounts of CKs that are most likely not plant-derived. This suggests that insects are able to provide CKs to the plant through their symbiotic association, thus extending further the role of insect bacterial symbionts in plant-insect interactions.}, } @article {pmid23802876, year = {2013}, author = {Le Clec'h, W and Raimond, M and Guillot, S and Bouchon, D and Sicard, M}, title = {Horizontal transfers of feminizing versus non-feminizing Wolbachia strains: from harmless passengers to pathogens.}, journal = {Environmental microbiology}, volume = {15}, number = {11}, pages = {2922-2936}, doi = {10.1111/1462-2920.12172}, pmid = {23802876}, issn = {1462-2920}, mesh = {Animals ; Isopoda/classification/*microbiology ; Phenotype ; *Symbiosis ; Virulence/genetics ; Wolbachia/classification/genetics/*pathogenicity ; }, abstract = {The endosymbiont Wolbachia pipientis infects various hosts in which it navigates vertically from mothers to offspring. However, horizontal transfers of Wolbachia can occur between hosts. The virulence of the horizontally acquired Wolbachia can change in the new host as it has been illustrated by the case of the feminizing strain wVulC from the woodlouse Armadillidium vulgare that turns to a pathogen when introduced into Porcellio dilatatus dilatatus. In the present study, we aim to show whether symbiotic traits, such as (i) host sex manipulation and (ii) colonization patterns, which differ between eight isopod Wolbachia strains, are connected to their virulence towards the recipient host P. d. dilatatus. Among the transferred Wolbachia, some feminizing strains gradually differing in feminizing intensity in their native hosts induced different levels of pathogenicity to P. d. dilatatus. Not a single feminizing strain passed vertically with high titres to the next generation. The non-feminizing Wolbachia strains, even if they reached high densities in the host, did not impact host life-history traits and some vertically passed with high titres to the offspring. These results suggest that a potential link between the manners Wolbachia manipulates its native host reproduction, its virulence and its ability to vertically infect the offspring.}, } @article {pmid23802459, year = {2013}, author = {Ritchie, SA and Montgomery, BL and Hoffmann, AA}, title = {Novel estimates of Aedes aegypti (Diptera: Culicidae) population size and adult survival based on Wolbachia releases.}, journal = {Journal of medical entomology}, volume = {50}, number = {3}, pages = {624-631}, doi = {10.1603/me12201}, pmid = {23802459}, issn = {0022-2585}, mesh = {Aedes/microbiology/*physiology ; Animals ; Ecology/*methods ; Entomology/*methods ; Female ; Male ; Pest Control, Biological/*methods ; Population Density ; Population Dynamics ; Queensland ; Wolbachia/physiology ; }, abstract = {The size of Aedes aegypti (L.) mosquito populations and adult survival rates have proven difficult to estimate because of a lack of consistent quantitative measures to equate sampling methods, such as adult trapping, to actual population size. However, such estimates are critical for devising control methods and for modeling the transmission of dengue and other infectious agents carried by this species. Here we take advantage of recent releases of Wolbachia-infected Ae. aegypti coupled with the results of ongoing monitoring to estimate the size of adult Ae. aegypti populations around Cairns in far north Queensland, Australia. Based on the association between released adults infected with Wolbachia and data from Biogents Sentinel traps, we show that data from two locations are consistent with population estimates of approximately 5-10 females per house and daily survival rates of 0.7-0.9 for the released Wolbachia-infected females. Moreover, we estimate that networks of Biogents Sentinel traps at a density of one per 15 houses capture around 5-10% of the adult population per week, and provide a rapid estimate of the absolute population size of Ae. aegypti. These data are discussed with respect to release rates and monitoring in future Wolbachia releases and also the levels of suppression required to reduce dengue transmission.}, } @article {pmid23799924, year = {2013}, author = {Zhang, KJ and Zhu, WC and Rong, X and Zhang, YK and Ding, XL and Liu, J and Chen, DS and Du, Y and Hong, XY}, title = {The complete mitochondrial genomes of two rice planthoppers, Nilaparvata lugens and Laodelphax striatellus: conserved genome rearrangement in Delphacidae and discovery of new characteristics of atp8 and tRNA genes.}, journal = {BMC genomics}, volume = {14}, number = {}, pages = {417}, pmid = {23799924}, issn = {1471-2164}, mesh = {Animals ; Base Composition ; Conserved Sequence/*genetics ; DNA, Mitochondrial/chemistry/genetics ; Gene Order/genetics ; Gene Rearrangement/*genetics ; Genes, Mitochondrial/genetics ; Genome, Mitochondrial/*genetics ; *Genomics ; Hemiptera/enzymology/*genetics ; Mitochondrial Proton-Translocating ATPases/*genetics ; Molecular Sequence Data ; RNA, Transfer/*genetics ; Regulatory Sequences, Nucleic Acid/genetics ; Species Specificity ; }, abstract = {BACKGROUND: Nilaparvata lugens (the brown planthopper, BPH) and Laodelphax striatellus (the small brown planthopper, SBPH) are two of the most important pests of rice. Up to now, there was only one mitochondrial genome of rice planthopper has been sequenced and very few dependable information of mitochondria could be used for research on population genetics, phylogeographics and phylogenetic evolution of these pests. To get more valuable information from the mitochondria, we sequenced the complete mitochondrial genomes of BPH and SBPH. These two planthoppers were infected with two different functional Wolbachia (intracellular endosymbiont) strains (wLug and wStri). Since both mitochondria and Wolbachia are transmitted by cytoplasmic inheritance and it was difficult to separate them when purified the Wolbachia particles, concomitantly sequencing the genome of Wolbachia using next generation sequencing method, we also got nearly complete mitochondrial genome sequences of these two rice planthoppers. After gap closing, we present high quality and reliable complete mitochondrial genomes of these two planthoppers.

RESULTS: The mitogenomes of N. lugens (BPH) and L. striatellus (SBPH) are 17, 619 bp and 16, 431 bp long with A + T contents of 76.95% and 77.17%, respectively. Both species have typical circular mitochondrial genomes that encode the complete set of 37 genes which are usually found in metazoans. However, the BPH mitogenome also possesses two additional copies of the trnC gene. In both mitochondrial genomes, the lengths of the atp8 gene were conspicuously shorter than that of all other known insect mitochondrial genomes (99 bp for BPH, 102 bp for SBPH). That two rearrangement regions (trnC-trnW and nad6-trnP-trnT) of mitochondrial genomes differing from other known insect were found in these two distantly related planthoppers revealed that the gene order of mitochondria might be conservative in Delphacidae. The large non-coding fragment (the A+T-rich region) putatively corresponding responsible for the control of replication and transcription of mitochondria contained a variable number of tandem repeats (VNTRs) block in different natural individuals of these two planthoppers. Comparison with a previously sequenced individual of SBPH revealed that the mitochondrial genetic variation within a species exists not only in the sequence and secondary structure of genes, but also in the gene order (the different location of trnH gene).

CONCLUSION: The mitochondrial genome arrangement pattern found in planthoppers was involved in rearrangements of both tRNA genes and protein-coding genes (PCGs). Different species from different genera of Delphacidae possessing the same mitochondrial gene rearrangement suggests that gene rearrangements of mitochondrial genome probably occurred before the differentiation of this family. After comparatively analyzing the gene order of different species of Hemiptera, we propose that except for some specific taxonomical group (e.g. the whiteflies) the gene order might have diversified in family level of this order. The VNTRs detected in the control region might provide additional genetic markers for studying population genetics, individual difference and phylogeographics of planthoppers.}, } @article {pmid23785694, year = {2013}, author = {Parvizi, P and Fardid, F and Soleimani, S}, title = {Detection of a New Strain of Wolbachia pipientis in Phlebotomus perfiliewi transcaucasicus, a Potential Vector of Visceral Leishmaniasis in North West of Iran, by Targeting the Major Surface Protein Gene.}, journal = {Journal of arthropod-borne diseases}, volume = {7}, number = {1}, pages = {46-55}, pmid = {23785694}, issn = {2322-1984}, abstract = {BACKGROUND: Wolbachia pipientis is maternally inherited endoparasitic bacterium belonging to the α-proteobacteria, infecting 20-75% of all insect species including sand flies. The Wolbachia surface protein (wsp) was employed as an appropriate marker for strain typing. The objective of our research was to find the possibility of detection of W. pipientis in Phlebotomus perfiliewi transcaucasicus.

METHODS: Individual sand flies were screened for the presence of W. pipientis. The obtained sequences were edited and aligned with database sequences to identify W. pipientis haplotypes.

RESULTS: Two haplotypes of W. pipientis were found in P. perfiliewi transcaucasicus. The common haplotype of W. pipientis was found to be identical to the sequences of those submitted in GenBank. New strain (haplotype) of W. pipientis was found novel. The sequence of new strain of W. pipientis occurs in P. perfiliewi transcaucasicus is very different from those already submitted in GenBank.

CONCLUSION: Finding one genetically modified new strain of W. pipientis in P. perfiliewi transcaucasicus, now we can conclude that further documents and studies need to reach the role of cytoplasmic incompatibility of W. pipientis through wild sand fly populations to drive a deleterious gene into and to reduce the density of natural populations of sand flies.}, } @article {pmid23778233, year = {2013}, author = {Nor, I and Engelstädter, J and Duron, O and Reuter, M and Sagot, MF and Charlat, S}, title = {On the genetic architecture of cytoplasmic incompatibility: inference from phenotypic data.}, journal = {The American naturalist}, volume = {182}, number = {1}, pages = {E15-24}, doi = {10.1086/670612}, pmid = {23778233}, issn = {1537-5323}, mesh = {Algorithms ; Alleles ; Animals ; Culex/*genetics/*microbiology/physiology ; Cytoplasm/microbiology ; Drosophila ; *Evolution, Molecular ; Female ; *Genome, Bacterial ; Hybridization, Genetic ; Male ; Models, Genetic ; Mutation ; Reproduction ; *Symbiosis ; Wolbachia/*genetics/*physiology ; }, abstract = {Numerous insects carry intracellular bacteria that manipulate the insects' reproduction and thus facilitate their own spread. Cytoplasmic incompatibility (CI) is a common form of such manipulation, where a (currently uncharacterized) bacterial modification of male sperm induces the early death of embryos unless the fertilized eggs carry the same bacteria, inherited from the mother. The death of uninfected embryos provides an indirect selective advantage to infected ones, thus enabling the spread of the bacteria. Here we use and expand recently developed algorithms to infer the genetic architecture underlying the complex incompatibility data from the mosquito Culex pipiens. We show that CI requires more genetic determinants than previously believed and that quantitative variation in gene products potentially contributes to the observed CI patterns. In line with population genetic theory of CI, our analysis suggests that toxin factors (those inducing embryo death) are present in fewer copies in the bacterial genomes than antitoxin factors (those ensuring that infected embryos survive). In combination with comparative genomics, our approach will provide helpful guidance to identify the genetic basis of CI and more generally of other toxin/antitoxin systems that can be conceptualized under the same framework.}, } @article {pmid23770905, year = {2013}, author = {Ishii, Y and Matsuura, Y and Kakizawa, S and Nikoh, N and Fukatsu, T}, title = {Diversity of bacterial endosymbionts associated with Macrosteles leafhoppers vectoring phytopathogenic phytoplasmas.}, journal = {Applied and environmental microbiology}, volume = {79}, number = {16}, pages = {5013-5022}, pmid = {23770905}, issn = {1098-5336}, mesh = {Animals ; Bacteria/*classification/*genetics/metabolism ; Bacterial Physiological Phenomena ; DNA, Bacterial/genetics/metabolism ; Female ; Hemiptera/*microbiology ; Japan ; Male ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics/metabolism ; Sequence Analysis, DNA ; Sequence Homology ; Species Specificity ; Symbiosis ; }, abstract = {Here, we investigate the endosymbiotic microbiota of the Macrosteles leafhoppers M. striifrons and M. sexnotatus, known as vectors of phytopathogenic phytoplasmas. PCR, cloning, sequencing, and phylogenetic analyses of bacterial 16S rRNA genes identified two obligate endosymbionts, "Candidatus Sulcia muelleri" and "Candidatus Nasuia deltocephalinicola," and five facultative endosymbionts, Wolbachia, Rickettsia, Burkholderia, Diplorickettsia, and a novel bacterium belonging to the Rickettsiaceae, from the leafhoppers. "Ca. Sulcia muelleri" and "Ca. Nasuia deltocephalinicola" exhibited 100% infection frequencies in the host species and populations and were separately harbored within different bacteriocytes that constituted a pair of coherent bacteriomes in the abdomen of the host insects, as in other deltocephaline leafhoppers. Wolbachia, Rickettsia, Burkholderia, Diplorickettsia, and the novel Rickettsiaceae bacterium exhibited infection frequencies at 7%, 31%, 12%, 0%, and 24% in M. striifrons and at 20%, 0%, 0%, 20%, and 0% in M. sexnotatus, respectively. Although undetected in the above analyses, phytoplasma infections were detected in 16% of M. striifrons and 60% of M. sexnotatus insects by nested PCR of 16S rRNA genes. Two genetically distinct phytoplasmas, namely, "Candidatus Phytoplasma asteris," associated with aster yellows and related plant diseases, and "Candidatus Phytoplasma oryzae," associated with rice yellow dwarf disease, were identified from the leafhoppers. These results highlight strikingly complex endosymbiotic microbiota of the Macrosteles leafhoppers and suggest ecological interactions between the obligate endosymbionts, the facultative endosymbionts, and the phytopathogenic phytoplasmas within the same host insects, which may affect vector competence of the leafhoppers.}, } @article {pmid23759724, year = {2013}, author = {Duron, O}, title = {Lateral transfers of insertion sequences between Wolbachia, Cardinium and Rickettsia bacterial endosymbionts.}, journal = {Heredity}, volume = {111}, number = {4}, pages = {330-337}, pmid = {23759724}, issn = {1365-2540}, mesh = {Animals ; Arthropods/genetics/microbiology ; Bacteria/genetics ; DNA Transposable Elements/*genetics ; Evolution, Molecular ; Gene Transfer, Horizontal/*genetics ; Molecular Sequence Data ; Phylogeny ; Rickettsia/*genetics ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Various bacteria live exclusively within arthropod cells and collectively act as an important driver of arthropod evolutionary ecology. Whereas rampant intra-generic DNA transfers were recently shown to have a pivotal role in the evolution of the most common of these endosymbionts, Wolbachia, the present study show that inter-generic DNA transfers also commonly take place, constituting a potent source of rapid genomic change. Bioinformatic, molecular and phylogenetic data provide evidence that a selfish genetic element, the insertion sequence ISRpe1, is widespread in the Wolbachia, Cardinium and Rickettsia endosymbionts and experiences recent (and likely ongoing) transfers over long evolutionary distances. Although many ISRpe1 copies were clearly expanding and leading to rapid endosymbiont diversification, degraded copies are also frequently found, constituting an unusual genomic fossil record suggestive of ancient ISRpe1 expansions. Overall, the present data highlight how ecological connections within the arthropod intracellular environment facilitate lateral DNA transfers between distantly related bacterial lineages.}, } @article {pmid23756849, year = {2013}, author = {Chan, MH and Kim, PS}, title = {Modelling a Wolbachia invasion using a slow-fast dispersal reaction-diffusion approach.}, journal = {Bulletin of mathematical biology}, volume = {75}, number = {9}, pages = {1501-1523}, doi = {10.1007/s11538-013-9857-y}, pmid = {23756849}, issn = {1522-9602}, mesh = {Aedes/microbiology/physiology ; Animals ; Computational Biology ; Drosophila/microbiology/physiology ; Female ; Host-Pathogen Interactions ; Male ; Mathematical Concepts ; *Models, Biological ; Population Dynamics ; Temperature ; Wolbachia/*pathogenicity ; }, abstract = {This paper uses a reaction-diffusion approach to examine the dynamics in the spread of a Wolbachia infection within a population of mosquitoes in a homogeneous environment. The formulated model builds upon an earlier model by Skalski and Gilliam (Am. Nat. 161(3):441-458, 2003), which incorporates a slow and fast dispersal mode. This generates a faster wavespeed than previous reaction-diffusion approaches, which have been found to produce wavespeeds that are unrealistically slow when compared with direct observations. In addition, the model incorporates cytoplasmic incompatibility between male and female mosquitoes, which creates a strong Allee effect in the dynamics. In previous studies, linearised wavespeeds have been found to be inaccurate when a strong Allee effect is underpinning the dynamics. We provide a means to approximate the wavespeed generated by the model and show that it is in close agreement with numerical simulations. Wavespeeds are approximated for both Aedes aegypti and Drosophila simulans mosquitoes at different temperatures. These wavespeeds indicate that as the temperature decreases within the optimal temperature range for mosquito survival, the speed of a Wolbachia invasion increases for Aedes aegypti populations and decreases for Drosophila simulans populations.}, } @article {pmid23755525, year = {2013}, author = {Serga, SV and Kozeretskaia, IA}, title = {[The puzzle of Wolbachia spreading out through natural populations of Drosophila melanogaster].}, journal = {Zhurnal obshchei biologii}, volume = {74}, number = {2}, pages = {99-111}, pmid = {23755525}, issn = {0044-4596}, mesh = {Animals ; Drosophila melanogaster/*genetics/*microbiology ; Fertility ; Host-Pathogen Interactions ; Population Dynamics ; Reproduction ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacteria Wolbachia are widely spread in natural populations of arthropods all over the world. This phenomenon, according to the current views, may be due either to so called reproductive parasitism (modification of sexual reproduction) or to mutualistic interrelationships with the host organism, since the bacteria are transmitted transovarially. Wolbachia are extremely widespread in natural populations and laboratory lineages of Drosophila melanogaster though they do not cause any known modifications of this insect sexual reproduction that could influence the bacteria frequency in the populations. It is found out that Wolbachia effects on drosophila reproductive processes can only consist in low-level of cytoplasmic incompatibility within laboratory lineages; in natural drosophila populations even such an effect has not been detected. The attempts to explain the wide spread of Wolbachia among representatives of D. melanogaster through fitness increasing of infected individuals were not successful either. Data obtained thereupon are contradictive and the results are not versatile. Meanwhile, there are some incomplete data on the bacteria affecting drosophila sexual behavior, expression of certain genes, ferrum metabolism. It is shown that the bacteria enhance individual resistance to RNA-viruses that, under natural conditions, infect fruit flies rather frequently, but the mechanisms of the phenomenon remain uncertain as well as the commonness of the phenomenon itself. Thus, at the one hand, it can be stated that infection by Wolbachia leads to a rather complex effect on D. melanogaster while, at the other hand, the reasons behind the wide spread of Wolbachia in natural populations of D. melanogaster still remain unclear.}, } @article {pmid23755311, year = {2013}, author = {Bian, G and Zhou, G and Lu, P and Xi, Z}, title = {Replacing a native Wolbachia with a novel strain results in an increase in endosymbiont load and resistance to dengue virus in a mosquito vector.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {6}, pages = {e2250}, pmid = {23755311}, issn = {1935-2735}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; R01 AI-080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*virology ; Animal Structures/virology ; Animals ; Bacterial Load ; Dengue Virus/*growth & development/*isolation & purification ; *Disease Vectors ; Microbial Interactions ; Viral Load ; Wolbachia/*growth & development ; }, abstract = {Wolbachia is a maternally transmitted endosymbiotic bacterium that is estimated to infect up to 65% of insect species. The ability of Wolbachia to both induce pathogen interference and spread into mosquito vector populations makes it possible to develop Wolbachia as a biological control agent for vector-borne disease control. Although Wolbachia induces resistance to dengue virus (DENV), filarial worms, and Plasmodium in mosquitoes, species like Aedes polynesiensis and Aedes albopictus, which carry native Wolbachia infections, are able to transmit dengue and filariasis. In a previous study, the native wPolA in Ae. polynesiensis was replaced with wAlbB from Ae. albopictus, and resulted in the generation of the transinfected "MTB" strain with low susceptibility for filarial worms. In this study, we compare the dynamics of DENV serotype 2 (DENV-2) within the wild type "APM" strain and the MTB strain of Ae. polynesiensis by measuring viral infection in the mosquito whole body, midgut, head, and saliva at different time points post infection. The results show that wAlbB can induce a strong resistance to DENV-2 in the MTB mosquito. Evidence also supports that this resistance is related to a dramatic increase in Wolbachia density in the MTB's somatic tissues, including the midgut and salivary gland. Our results suggests that replacement of a native Wolbachia with a novel infection could serve as a strategy for developing a Wolbachia-based approach to target naturally infected insects for vector-borne disease control.}, } @article {pmid23744038, year = {2013}, author = {Toomey, ME and Panaram, K and Fast, EM and Beatty, C and Frydman, HM}, title = {Evolutionarily conserved Wolbachia-encoded factors control pattern of stem-cell niche tropism in Drosophila ovaries and favor infection.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {110}, number = {26}, pages = {10788-10793}, pmid = {23744038}, issn = {1091-6490}, support = {K22 AI074909/AI/NIAID NIH HHS/United States ; 1K22AI74909-01A1/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics/physiology ; Drosophila/genetics/*microbiology ; Female ; Host-Pathogen Interactions ; Male ; Ovary/cytology/*microbiology ; Ovum/microbiology ; Phylogeny ; Species Specificity ; Stem Cell Niche ; Stem Cells/*microbiology ; Symbiosis ; Wolbachia/*genetics/pathogenicity/*physiology ; }, abstract = {Wolbachia are intracellular bacteria that infect invertebrates at pandemic levels, including insect vectors of devastating infectious diseases. Although Wolbachia are providing novel strategies for the control of several human pathogens, the processes underlying Wolbachia's successful propagation within and across species remain elusive. Wolbachia are mainly vertically transmitted; however, there is also evidence of extensive horizontal transmission. Here, we provide several lines of evidence supporting Wolbachia's targeting of ovarian stem cell niches--referred to as "niche tropism"--as a previously overlooked strategy for Wolbachia thriving in nature. Niche tropism is pervasive in Wolbachia infecting the Drosophila genus, and different patterns of niche tropism are evolutionarily conserved. Phylogenetic analysis, confirmed by hybrid introgression and transinfection experiments, demonstrates that bacterial factors are the major determinants of differential patterns of niche tropism. Furthermore, bacterial load is increased in germ-line cells passing through infected niches, supporting previous suggestions of a contribution of Wolbachia from stem-cell niches toward vertical transmission. These results support the role of stem-cell niches as a key component for the spreading of Wolbachia in the Drosophila genus and provide mechanistic insights into this unique tissue tropism.}, } @article {pmid23733960, year = {2013}, author = {Zhang, G and Hussain, M and O'Neill, SL and Asgari, S}, title = {Wolbachia uses a host microRNA to regulate transcripts of a methyltransferase, contributing to dengue virus inhibition in Aedes aegypti.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {110}, number = {25}, pages = {10276-10281}, pmid = {23733960}, issn = {1091-6490}, mesh = {Aedes/genetics/*microbiology/*virology ; Animals ; Cell Line ; DNA (Cytosine-5-)-Methyltransferases/*genetics/metabolism ; Dengue/*microbiology/transmission/virology ; Dengue Virus/*genetics/growth & development ; Female ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Enzymologic ; Insect Vectors/genetics/microbiology/virology ; MicroRNAs/genetics ; Ovary/enzymology/microbiology/virology ; Symbiosis/physiology ; Virus Replication/physiology ; Wolbachia/growth & development/*virology ; }, abstract = {The endosymbiont Wolbachia is common among insects and known for the reproductive manipulations it exerts on hosts as well as inhibition of virus replication in their hosts. Recently, we showed that Wolbachia uses host microRNAs to manipulate host gene expression for its efficient maintenance in the dengue mosquito vector, Aedes aegypti. Cytosine methylation is mediated by a group of proteins called DNA (cytosine-5) methyltransferases, which are structurally and functionally conserved from prokaryotes to eukaryotes. The biological functions of cytosine methylation include host defense, genome stability, gene regulation, developmental promotion of organs, and lifespan regulation. Ae. aegypti has only one DNA methyltransferase gene (AaDnmt2) belonging to the cytosine methyltransferase family 2, which is the most deeply conserved and widely distributed gene among metazoans. Here, we show that in mosquitoes the introduced endosymbiont, Wolbachia, significantly suppresses expression of AaDnmt2, but dengue virus induces expression of AaDnmt2. Interestingly, we found that aae-miR-2940 microRNA, which is exclusively expressed in Wolbachia-infected mosquitoes, down-regulates the expression of AaDnmt2. Reversely, overexpression of AaDnmt2 in mosquito cells led to inhibition of Wolbachia replication, but significantly promoted replication of dengue virus, suggesting a causal link between this Wolbachia manipulation and the blocking of dengue replication in Wolbachia-infected mosquitoes. In addition, our findings provide an explanation for hypomethylation of the genome in Wolbachia-infected Ae. aegypti.}, } @article {pmid23733469, year = {2013}, author = {Rana, AK and Chandra, S and Siddiqi, MI and Misra-Bhattacharya, S}, title = {Molecular characterization of an rsmD-like rRNA methyltransferase from the Wolbachia endosymbiont of Brugia malayi and antifilarial activity of specific inhibitors of the enzyme.}, journal = {Antimicrobial agents and chemotherapy}, volume = {57}, number = {8}, pages = {3843-3856}, pmid = {23733469}, issn = {1098-6596}, mesh = {Animals ; Brugia malayi/drug effects/genetics/*microbiology ; Cloning, Molecular ; Culicidae ; Disease Models, Animal ; Drug Evaluation, Preclinical ; Enzyme Inhibitors/pharmacology ; Female ; Filaricides/administration & dosage/*pharmacology ; Genes, Bacterial ; Gerbillinae ; Inhibitory Concentration 50 ; Male ; Methyltransferases/*antagonists & inhibitors/genetics/*isolation & purification/metabolism ; Murinae ; Substrate Specificity ; Symbiosis ; Tryptophan/metabolism ; Wolbachia/*enzymology/growth & development ; }, abstract = {The endosymbiotic organism Wolbachia is an attractive antifilarial drug target. Here we report on the cloning and expression of an rsmD-like rRNA methyltransferase from the Wolbachia endosymbiont of Brugia malayi, its molecular properties, and assays for specific inhibitors. The gene was found to be expressed in all the major life stages of B. malayi. The purified enzyme expressed in Escherichia coli was found to be in monomer form in its native state. The activities of the specific inhibitors (heteroaryl compounds) against the enzyme were tested with B. malayi adult and microfilariae for 7 days in vitro at various concentrations, and NSC-659390 proved to be the most potent compound (50% inhibitory concentration [IC50], 0.32 μM), followed by NSC-658343 (IC50, 4.13 μM) and NSC-657589 (IC50, 7.5 μM). On intraperitoneal administration at 5 mg/kg of body weight for 7 days to adult jirds into which B. malayi had been transplanted intraperitoneally, all the compounds killed a significant proportion of the implanted worms. A very similar result was observed in infected mastomys when inhibitors were administered. Docking studies of enzyme and inhibitors and an in vitro tryptophan quenching experiment were also performed to understand the binding mode and affinity. The specific inhibitors of the enzyme showed a higher affinity for the catalytic site of the enzyme than the nonspecific inhibitors and were found to be potent enough to kill the worm (both adults and microfilariae) in vitro as well as in vivo in a matter of days at micromolar concentrations. The findings suggest that these compounds be evaluated against other pathogens possessing a methyltransferase with a DPPY motif and warrant the design and synthesis of more such inhibitors.}, } @article {pmid23726053, year = {2013}, author = {Zhao, DX and Zhang, XF and Hong, XY}, title = {Host-symbiont interactions in spider mite Tetranychus truncates doubly infected with Wolbachia and Cardinium.}, journal = {Environmental entomology}, volume = {42}, number = {3}, pages = {445-452}, doi = {10.1603/EN12354}, pmid = {23726053}, issn = {1938-2936}, mesh = {Animals ; Bacteroidetes/*physiology ; China ; Female ; Fertility ; Longevity ; Male ; Reproduction ; *Symbiosis ; Tetranychidae/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Inherited bacteria Wolbachia, and more recently Cardinium, have received a great deal of attention for their ability to manipulate the reproduction and fitness of their host species. Wolbachia and Cardinium have been found to co-infect the same host species. In this study, both Wolbachia and Cardinium were found to manipulate host reproduction through cytoplasmic incompatibility and to affect the male-biased sex-ratio in the doubly infected spider mite Tetranychus truncates Ehara. We also investigated effects of double infection with Wolbachia and Cardinium on host fecundity and longevity. Results indicated that Wolbachia and Cardinium increased the fecundity of doubly infected females, although no infection effect on host longevity was observed. Our most important finding was that the mating of uninfected mites facilitated the proliferation of Wolbachia and Cardinium in double-infected mites. We discuss the results observed with respect to the spread of bacterial infection in natural populations and the evolution of the endosymbiont-T. truncates symbiosis.}, } @article {pmid23719839, year = {2013}, author = {Fallon, AM and Kurtz, CM and Carroll, EM}, title = {The oxidizing agent, paraquat, is more toxic to Wolbachia than to mosquito host cells.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {49}, number = {7}, pages = {501-507}, pmid = {23719839}, issn = {1543-706X}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/cytology/*drug effects/microbiology ; Amino Acid Sequence ; Animals ; Antioxidants/metabolism ; Bacterial Proteins/genetics/metabolism ; Cell Line ; Dose-Response Relationship, Drug ; Genome, Bacterial ; Herbicides/*toxicity ; Host-Pathogen Interactions ; Molecular Sequence Data ; Oxidants/*toxicity ; Paraquat/*toxicity ; Polymerase Chain Reaction ; Superoxide Dismutase/genetics/metabolism ; Wolbachia/*drug effects/growth & development ; }, abstract = {Cultured cells provide an important in vitro system for examining metabolic interactions between the intracellular bacterium, Wolbachia pipientis, and its insect hosts. To test whether Wolbachia-associated changes in antioxidant activities could provide a tool to select for infected cells, we tested the effects of paraquat (PQ) on Aedes albopictus mosquito cells. Like mammalian cells, mosquito cells tolerate PQ over a wide range of concentrations, and for considerable lengths of time, depending on cell density at the time of treatment. When mosquito cells were plated at low density and allowed to grow in the presence of PQ, we measured an LC50 of approximately 1-2 μM. Unexpectedly, cells persistently infected with Wolbachia strain wStr, from the planthopper Laodelphax striatellus, grew to higher densities in the presence of 1.5 μM PQ than in its absence. This effect of PQ was similar to the improved growth of host cells that occurs in the presence of antibiotics that suppress the Wolbachia infection. A more detailed examination of growth and metabolic sensitivity indicated that wStr is about 10-fold more sensitive to PQ than the mosquito host cells. Microscopic examination confirmed that Wolbachia levels were reduced in PQ-treated cells, and DNA estimates based on the polymerase chain reaction (PCR) indicated that Wolbachia abundance decreased by approximately 100-fold over a 10-d period. Although Wolbachia genomes encode superoxide dismutase, inspection of annotated genomes indicates that they lack several genes encoding products that ameliorate oxidative damage, including catalase, which converts the PQ byproduct, hydrogen peroxide, to molecular oxygen and water. We suggest that loss of multiple genes that participate in repair of oxidative damage accounts for increased sensitivity of Wolbachia to PQ, relative to its host cells.}, } @article {pmid23716403, year = {2013}, author = {Yeap, HL and Endersby, NM and Johnson, PH and Ritchie, SA and Hoffmann, AA}, title = {Body size and wing shape measurements as quality indicators of Aedes aegypti mosquitoes destined for field release.}, journal = {The American journal of tropical medicine and hygiene}, volume = {89}, number = {1}, pages = {78-92}, pmid = {23716403}, issn = {1476-1645}, mesh = {Aedes/*anatomy & histology/microbiology ; Animals ; Body Size ; Female ; Male ; Mosquito Control ; Population Dynamics ; Queensland ; Wings, Animal/*anatomy & histology ; Wolbachia ; }, abstract = {There is increasing interest in rearing modified mosquitoes for mass release to control vector-borne diseases, particularly Wolbachia-infected Aedes aegypti for suppression of dengue. Successful introductions require release of high quality mosquitoes into natural populations. Potential indicators of quality are body size and shape. We tested to determine if size, wing/thorax ratio, and wing shape are associated with field fitness of Wolbachia-infected Ae. aegypti. Compared with field-collected mosquitoes, released mosquitoes were larger in size, with lower size variance and different wing shape but similar in wing-thorax ratio and its associated variance. These differences were largely attributed to nutrition and to a minor extent to wMel Wolbachia infection. Survival potential of released female mosquitoes was similar to those from the field. Females at oviposition sites tended to be larger than those randomly collected from BG-Sentinel traps. Rearing conditions should thus aim for large size without affecting wing/thorax ratios.}, } @article {pmid23701629, year = {2013}, author = {Tay, ST}, title = {Wolbachia endosymbionts, Rickettsia felis and Bartonella species, in Ctenocephalides felis fleas in a tropical region.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {38}, number = {1}, pages = {200-202}, doi = {10.1111/j.1948-7134.2013.12030.x}, pmid = {23701629}, issn = {1948-7134}, mesh = {Animals ; Ctenocephalides/*microbiology ; Rickettsia felis/*physiology ; Tropical Climate ; Wolbachia/*physiology ; }, } @article {pmid23681021, year = {2013}, author = {Endersby, NM and White, VL and Chan, J and Hurst, T and Rašić, G and Miller, A and Hoffmann, AA}, title = {Evidence of cryptic genetic lineages within Aedes notoscriptus (Skuse).}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {18}, number = {}, pages = {191-201}, doi = {10.1016/j.meegid.2013.04.035}, pmid = {23681021}, issn = {1567-7257}, mesh = {Aedes/classification/*genetics/microbiology ; Animals ; Australia ; Bayes Theorem ; Gene Pool ; Genetic Markers/genetics ; Insect Vectors/classification/*genetics/microbiology ; New Zealand ; Phylogeny ; Rickettsiaceae Infections/microbiology/transmission ; Sequence Analysis, DNA ; Wolbachia/isolation & purification ; }, abstract = {Aedes notoscriptus (Skuse), a mosquito from the southwest Pacific region including Australia, has been implicated as a vector of arboviruses, but its status as a species is unclear. To investigate the taxonomic situation, we assessed genetic variation and phylogenetic relationships among Ae. notoscriptus from the east coast of Australia, Western Australia and New Zealand. Phylogenetic analyses of DNA sequence data from mitochondrial markers indicate that Ae. notoscriptus is a complex of divergent genetic lineages, some of which appear geographically restricted, while others are widespread in eastern Australia. Samples from New Zealand and Western Australia were related to populations from one southern Australian lineage. Nuclear markers show no evidence of genetic isolation by geographic distance in the overall sample of mosquitoes, but strong isolation by distance is obvious within two of the lineages, supporting their status as isolated gene pools. The morphological character of wing centroid size variation is also associated with genetic lineage. These findings point to the possibility that Ae. notoscriptus is a complex of species, highlighting the need to understand physiological and ecological differences that may influence future control strategies.}, } @article {pmid23672808, year = {2013}, author = {Strunov, A and Kiseleva, E and Gottlieb, Y}, title = {Spatial and temporal distribution of pathogenic Wolbachia strain wMelPop in Drosophila melanogaster central nervous system under different temperature conditions.}, journal = {Journal of invertebrate pathology}, volume = {114}, number = {1}, pages = {22-30}, doi = {10.1016/j.jip.2013.05.001}, pmid = {23672808}, issn = {1096-0805}, mesh = {Animals ; Central Nervous System/microbiology ; Drosophila melanogaster/growth & development/*microbiology ; Embryonic Development ; In Situ Hybridization, Fluorescence ; Larva/microbiology ; *Temperature ; Wolbachia/classification/*isolation & purification ; }, abstract = {The pathogenic Wolbachia strain, wMelPop, of Drosophila melanogaster is propagated in the fly's brain and muscles. To determine how wMelPop spreads in the host's central nervous system (CNS) during its life cycle, we used whole-mount fluorescent in situ hybridization to demonstrate the spatial distribution of wMelPop in D.melanogaster larvae and adults. To assess the effect of temperature on the pattern of wMelPop spread, we performed this analysis under moderate (25°C) and high (29°C) temperature conditions. Wolbachia distribution pattern in the third instar larva and adult brain was similar at both temperatures. wMelPop was generally localized to the subesophageal ganglion and the central brain of the host, whereas optic lobe anlagen cells of third instar larvae and cells of the optic lobe, lamina and retina of adult flies were mostly free of bacteria. Interestingly, high temperature had no significant effect on wMelPop titer or localization in the brain during larval development, but considerably altered it in adults immediately after eclosion. At both temperatures and within all tested stages of the life cycle, the bacterial titer varied only slightly between individuals. The observed differences in wMelPop titers in the central brain, subesophageal ganglion and optic lobe anlagen cells of third instar larva's CNS, together with the observation that these patterns are conserved in the adult brain, suggest that Wolbachia distribution is determined during fly embryogenesis.}, } @article {pmid23671034, year = {2013}, author = {Faria, VG and Sucena, E}, title = {Wolbachia in the Malpighian tubules: evolutionary dead-end or adaptation?.}, journal = {Journal of experimental zoology. Part B, Molecular and developmental evolution}, volume = {320}, number = {4}, pages = {195-199}, doi = {10.1002/jez.b.22498}, pmid = {23671034}, issn = {1552-5015}, mesh = {Adaptation, Physiological/*genetics ; Animals ; *Biological Evolution ; Drosophila/growth & development/microbiology ; Embryonic Development/genetics ; Germ Cells/microbiology ; Malpighian Tubules/cytology/*microbiology ; Symbiosis/genetics ; Wolbachia/*genetics/physiology ; }, abstract = {Facultative endosymbionts, such as Wolbachia, perpetuate by vertical transmission mostly through colonization of the germline during embryogenesis. The remaining Wolbachia inside the embryo are internalized in progenitor cells of the somatic tissue. This perpetuation strategy triggers a cyclic bacterial bottleneck across host generations. However, throughout the host's life history (Drosophila, for example), some somatic tissues such as the Malpighian tubules (MTs) show large numbers of Wolbachia. It is assumed that Wolbachia present in the progenitor cells of the MTs are confined to this somatic tissue, implicitly considering MTs as an evolutionary dead-end for these bacteria. Nevertheless, the fact that bacteria can survive and proliferate inside MTs suggests a different fate as they may access the host's reproductive system and persist in the host population through vertical transmission. Indeed, based on the particular physiological and developmental characteristics of MT, as well as of Wolbachia, we argue the bacteria present in the MTs may constitute a secondary pool of vertically transmitted bacteria. Moreover, somatic pools of Wolbachia capable of reaching the gonads and insure vertical transmission may also provide an interesting element to the elucidation of horizontal transmission mechanisms. Finally, we also speculate that somatic pools of Wolbachia may play an important role in host fitness, namely during viral infections. In brief, we argue that the somatic pools of Wolbachia, with special emphasis on the MT subset, deserve experimental attention as putative players in the physiology and evolution of both bacteria and hosts.}, } @article {pmid23662926, year = {2013}, author = {Rasheed, SB and Boots, M and Frantz, AC and Butlin, RK}, title = {Population structure of the mosquito Aedes aegypti (Stegomyia aegypti) in Pakistan.}, journal = {Medical and veterinary entomology}, volume = {27}, number = {4}, pages = {430-440}, doi = {10.1111/mve.12001}, pmid = {23662926}, issn = {1365-2915}, mesh = {Aedes/*genetics/*physiology ; Animals ; Demography ; Genetic Variation ; Microsatellite Repeats/*genetics ; Pakistan ; Software ; }, abstract = {Eleven microsatellite markers were used to determine the genetic population structure and spread of Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) in Pakistan using mosquitoes collected from 13 different cities. There is a single genetic cluster of Ae. aegypti in Pakistan with a pattern of isolation by distance within the population. The low level of isolation by distance suggests the long-range passive dispersal of this mosquito, which may be facilitated by the tyre trade in Pakistan. A decrease in genetic diversity from south to north suggests a recent spread of this mosquito from Karachi. A strong negative correlation between genetic distance and the quality of road connections shows that populations in cities connected by better road networks are less differentiated, which suggests the human-aided passive dispersal of Ae. aegypti in Pakistan. Dispersal on a large spatial scale may facilitate the strategy of introducing transgenic Ae. aegypti or intracellular bacteria such as Wolbachia to control the spread of dengue disease in Pakistan, but it also emphasizes the need for simple measures to control container breeding sites.}, } @article {pmid23661760, year = {2013}, author = {Bian, G and Joshi, D and Dong, Y and Lu, P and Zhou, G and Pan, X and Xu, Y and Dimopoulos, G and Xi, Z}, title = {Wolbachia invades Anopheles stephensi populations and induces refractoriness to Plasmodium infection.}, journal = {Science (New York, N.Y.)}, volume = {340}, number = {6133}, pages = {748-751}, doi = {10.1126/science.1236192}, pmid = {23661760}, issn = {1095-9203}, support = {R01AI061576/AI/NIAID NIH HHS/United States ; R01AI080597/AI/NIAID NIH HHS/United States ; R21AI082141/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology ; Female ; Humans ; Malaria, Falciparum/parasitology/*prevention & control ; Male ; *Pest Control, Biological ; Plasmodium falciparum/*growth & development ; Reactive Oxygen Species/metabolism ; Wolbachia/*growth & development ; }, abstract = {Wolbachia is a maternally transmitted symbiotic bacterium of insects that has been proposed as a potential agent for the control of insect-transmitted diseases. One of the major limitations preventing the development of Wolbachia for malaria control has been the inability to establish inherited infections of Wolbachia in anopheline mosquitoes. Here, we report the establishment of a stable Wolbachia infection in an important malaria vector, Anopheles stephensi. In A. stephensi, Wolbachia strain wAlbB displays both perfect maternal transmission and the ability to induce high levels of cytoplasmic incompatibility. Seeding of naturally uninfected A. stephensi populations with infected females repeatedly resulted in Wolbachia invasion of laboratory mosquito populations. Furthermore, wAlbB conferred resistance in the mosquito to the human malaria parasite Plasmodium falciparum.}, } @article {pmid23660513, year = {2013}, author = {Wiwatanaratanabutr, I}, title = {Geographic distribution of wolbachial infections in mosquitoes from Thailand.}, journal = {Journal of invertebrate pathology}, volume = {114}, number = {3}, pages = {337-340}, doi = {10.1016/j.jip.2013.04.011}, pmid = {23660513}, issn = {1096-0805}, mesh = {Aedes/microbiology ; Animals ; Culex/microbiology ; Culicidae/*microbiology ; DNA, Bacterial/chemistry ; Geography ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Thailand ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Members of the genus Wolbachia are inherited intracellular bacterial endosymbionts that infect a diverse range of arthropods. Here I report the results of a survey of these endosymbionts in different mosquito species from six geographic regions of Northern, Northeastern, Western, Central, Eastern and Southern Thailand. Using gene amplification assays with wsp and groE gene primers, wolbachiae were detected in 999 mosquitoes representing 28 species of 1622 specimens collected representing 74 species of wild-caught mosquitoes from all regions of Thailand. Results using wsp primers were similar to those using groE primers in all cases. Wolbachiae had not been reported previously from five of the species tested, namely, Aedes lineatopennis, Aedes vexans, Aedes vittatus, Culex pallidothorax and Culex whitmorei. Infections were found in all major disease vector genera except Anopheles. These results indicate that wolbachial infections are distributed throughout many mosquito species in Thailand.}, } @article {pmid23645190, year = {2013}, author = {Iasur-Kruh, L and Weintraub, PG and Mozes-Daube, N and Robinson, WE and Perlman, SJ and Zchori-Fein, E}, title = {Novel Rickettsiella bacterium in the leafhopper Orosius albicinctus (Hemiptera: Cicadellidae).}, journal = {Applied and environmental microbiology}, volume = {79}, number = {14}, pages = {4246-4252}, pmid = {23645190}, issn = {1098-5336}, support = {T32 CA009110/CA/NCI NIH HHS/United States ; CA-9110-09/CA/NCI NIH HHS/United States ; }, mesh = {Animals ; Bacteroidetes/classification/genetics/physiology ; Coxiellaceae/*classification/genetics/*isolation & purification/physiology ; DNA, Bacterial/genetics ; Denaturing Gradient Gel Electrophoresis ; Enterobacteriaceae/classification/genetics/physiology ; Female ; Hemiptera/*microbiology/*physiology ; In Situ Hybridization, Fluorescence ; Israel ; Male ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics/metabolism ; Sequence Alignment ; Sequence Analysis, DNA ; Symbiosis ; }, abstract = {Bacteria in the genus Rickettsiella (Coxiellaceae), which are mainly known as arthropod pathogens, are emerging as excellent models to study transitions between mutualism and pathogenicity. The current report characterizes a novel Rickettsiella found in the leafhopper Orosius albicinctus (Hemiptera: Cicadellidae), a major vector of phytoplasma diseases in Europe and Asia. Denaturing gradient gel electrophoresis (DGGE) and pyrosequencing were used to survey the main symbionts of O. albicinctus, revealing the obligate symbionts Sulcia and Nasuia, and the facultative symbionts Arsenophonus and Wolbachia, in addition to Rickettsiella. The leafhopper Rickettsiella is allied with bacteria found in ticks. Screening O. albicinctus from the field showed that Rickettsiella is highly prevalent, with over 60% of individuals infected. A stable Rickettsiella infection was maintained in a leafhopper laboratory colony for at least 10 generations, and fluorescence microscopy localized bacteria to accessory glands of the female reproductive tract, suggesting that the bacterium is vertically transmitted. Future studies will be needed to examine how Rickettsiella affects host fitess and its ability to vector phytopathogens.}, } @article {pmid23634597, year = {2013}, author = {Mains, JW and Brelsfoard, CL and Crain, PR and Huang, Y and Dobson, SL}, title = {Population impacts of Wolbachia on Aedes albopictus.}, journal = {Ecological applications : a publication of the Ecological Society of America}, volume = {23}, number = {2}, pages = {493-501}, doi = {10.1890/12-1097.1}, pmid = {23634597}, issn = {1051-0761}, support = {AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Female ; Host-Pathogen Interactions ; Male ; Models, Biological ; Population Density ; Wolbachia/*physiology ; }, abstract = {Prior studies have demonstrated that Wolbachia, a commonly occurring bacterium capable of manipulating host reproduction, can affect life history traits in insect hosts, which in turn can have population-level effects. Effects on hosts at the individual level are predicted to impact population dynamics, but the latter has not been examined empirically. Here, we describe a biological model system based on Aedes albopictus (Asian tiger mosquito) that allows for measurement of population dynamics, which has not been accomplished in prior field trials or laboratory designs. The results demonstrate the studied populations to be robust and allow for persistent, closed populations with overlapping generations, which are regulated solely through density-dependent, intraspecific competition for limited resources. Using a novel experimental design, we compare populations that are either uninfected or infected with Wolbachia. The results show differences that include population size, eclosion rates, adult survivorship, and fecundity. The aposymbiotic populations were generally larger and adults longer lived relative to the infected populations. The outcome is discussed in context with naturally occurring Wolbachia invasions, proposed autocidal strategies, and the utility of the developed system as a biological platform for hypothesis testing and improved parameterization.}, } @article {pmid23623993, year = {2013}, author = {Rodriguero, MS and Lanteri, AA and Confalonieri, VA}, title = {Speciation in the asexual realm: is the parthenogenetic weevil Naupactus cervinus a complex of species in statu nascendi?.}, journal = {Molecular phylogenetics and evolution}, volume = {68}, number = {3}, pages = {644-656}, doi = {10.1016/j.ympev.2013.04.011}, pmid = {23623993}, issn = {1095-9513}, mesh = {Animals ; Argentina ; *Biological Evolution ; Brazil ; DNA, Mitochondrial/genetics ; Female ; Genetic Variation ; Genotype ; Multilocus Sequence Typing ; *Parthenogenesis ; Phenotype ; Phylogeny ; Weevils/*classification/*physiology ; }, abstract = {Population genetic theory shows that asexual organisms may evolve into species, which behave as independent evolutionary units. As a result, they form genotypic clusters separated by deep gaps due to geographic isolation and/or divergent selection. Identification of several genetically divergent groups of weevils embodied in the nominal species Naupactus cervinus deserves further study, in order to test if these lineages are evolving independently. In the present paper we tested if the parthenogenetic weevil N. cervinus, native to South America and broadly distributed throughout the world, contains more than one evolutionary unit. For this purpose, we applied three different approaches, a multilocus phylogenetic analysis, the GMYC approach and the K/θ method. We accomplished these analyses through a survey of mitochondrial (COI and COII genes) and nuclear (ITS1 sequence) genetic variation and morphometric analysis in a sample which included individuals from different locations within the native geographic range of N. cervinus. In addition, we compared the divergence accumulated in this species with that in another weevil of the same tribe (Naupactini) showing identical reproductive mode to see if similar levels of morphological variation matches similar levels of genetic divergence. We report the presence of two independent evolutionary units living in sympatry in forest areas. The incongruence between mitochondrial and nuclear datasets analyzed herein reflects incomplete lineage sorting of the nuclear marker and different evolutionary rates between genomes. Ecological divergence driven by natural selection (sympatry) or secondary contact after geographic isolation (allopatry) might explain the deep gaps in mitochondrial phylogenies. Instead, Wolbachia infection was ruled out as a causal factor for such differentiation. We conclude that N. cervinus is probably a species complex with at least two well differentiated lineages that would represent a cluster of species in statu nascendi.}, } @article {pmid23613801, year = {2013}, author = {Boucias, DG and Kariithi, HM and Bourtzis, K and Schneider, DI and Kelley, K and Miller, WJ and Parker, AG and Abd-Alla, AM}, title = {Transgenerational transmission of the Glossina pallidipes hytrosavirus depends on the presence of a functional symbiome.}, journal = {PloS one}, volume = {8}, number = {4}, pages = {e61150}, pmid = {23613801}, issn = {1932-6203}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Ampicillin/pharmacology ; Animals ; Female ; Gram-Negative Bacteria/*physiology ; Insect Viruses/drug effects/*physiology ; Male ; *Symbiosis ; Tsetse Flies/*microbiology/*virology ; Virus Replication/drug effects ; }, abstract = {The vertically transmitted endosymbionts (Sodalis glossinidius and Wigglesworthia glossinidia) of the tsetse fly (Diptera: Glossinidae) are known to supplement dietary deficiencies and modulate the reproductive fitness and the defense system of the fly. Some tsetse fly species are also infected with the bacterium, Wolbachia and with the Glossina hytrosavirus (GpSGHV). Laboratory-bred G. pallidipes exhibit chronic asymptomatic and acute symptomatic GpSGHV infection, with the former being the most common in these colonies. However, under as yet undefined conditions, the asymptomatic state can convert to the symptomatic state, leading to detectable salivary gland hypertrophy (SGH(+)) syndrome. In this study, we investigated the interplay between the bacterial symbiome and GpSGHV during development of G. pallidipes by knocking down the symbionts with antibiotic. Intrahaemocoelic injection of GpSGHV led to high virus titre (10(9) virus copies), but was not accompanied by either the onset of detectable SGH(+), or release of detectable virus particles into the blood meals during feeding events. When the F1 generations of GpSGHV-challenged mothers were dissected within 24 h post-eclosion, SGH(+) was observed to increase from 4.5% in the first larviposition cycle to >95% in the fourth cycle. Despite being sterile, these F1 SGH(+) progeny mated readily. Removal of the tsetse symbiome, however, suppressed transgenerational transfer of the virus via milk secretions and blocked the ability of GpSGHV to infect salivary glands of the F1 progeny. Whereas GpSGHV infects and replicates in salivary glands of developing pupa, the virus is unable to induce SGH(+) within fully differentiated adult salivary glands. The F1 SGH(+) adults are responsible for the GpSGHV-induced colony collapse in tsetse factories. Our data suggest that GpSGHV has co-evolved with the tsetse symbiome and that the symbionts play key roles in the virus transmission from mother to progeny.}, } @article {pmid23593519, year = {2013}, author = {Melnikow, E and Xu, S and Liu, J and Bell, AJ and Ghedin, E and Unnasch, TR and Lustigman, S}, title = {A potential role for the interaction of Wolbachia surface proteins with the Brugia malayi glycolytic enzymes and cytoskeleton in maintenance of endosymbiosis.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {4}, pages = {e2151}, pmid = {23593519}, issn = {1935-2735}, support = {R01 AI072465/AI/NIAID NIH HHS/United States ; AI072465/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/metabolism ; Brugia malayi/*metabolism/*microbiology ; Cytoskeleton/*metabolism ; Membrane Proteins/*metabolism ; Protozoan Proteins/metabolism ; Symbiosis/*physiology ; Wolbachia/*metabolism/*physiology ; }, abstract = {The human filarial parasite Brugia malayi harbors an endosymbiotic bacterium of the genus Wolbachia. The Wolbachia represent an attractive target for the control of filarial induced disease as elimination of the bacteria affects molting, reproduction and survival of the worms. The molecular basis for the symbiotic relationship between Wolbachia and their filarial hosts has yet to be elucidated. To identify proteins involved in this process, we focused on the Wolbachia surface proteins (WSPs), which are known to be involved in bacteria-host interactions in other bacterial systems. Two WSP-like proteins (wBm0152 and wBm0432) were localized to various host tissues of the B. malayi female adult worms and are present in the excretory/secretory products of the worms. We provide evidence that both of these proteins bind specifically to B. malayi crude protein extracts and to individual filarial proteins to create functional complexes. The wBm0432 interacts with several key enzymes involved in the host glycolytic pathway, including aldolase and enolase. The wBm0152 interacts with the host cytoskeletal proteins actin and tubulin. We also show these interactions in vitro and have verified that wBm0432 and B. malayi aldolase, as well as wBm0152 and B. malayi actin, co-localize to the vacuole surrounding Wolbachia. We propose that both WSP protein complexes interact with each other via the aldolase-actin link and/or via the possible interaction between the host's enolase and the cytoskeleton, and play a role in Wolbachia distribution during worm growth and embryogenesis.}, } @article {pmid23593179, year = {2013}, author = {Le Clec'h, W and Chevalier, FD and Genty, L and Bertaux, J and Bouchon, D and Sicard, M}, title = {Cannibalism and predation as paths for horizontal passage of Wolbachia between terrestrial isopods.}, journal = {PloS one}, volume = {8}, number = {4}, pages = {e60232}, pmid = {23593179}, issn = {1932-6203}, mesh = {Animals ; Base Sequence ; *Cannibalism ; DNA Primers ; *Gene Transfer, Horizontal ; In Situ Hybridization, Fluorescence ; Phylogeny ; Polymerase Chain Reaction ; *Predatory Behavior ; Wolbachia/classification/genetics/*physiology ; }, abstract = {The alpha-proteobacteria Wolbachia are the most widespread endosymbionts in arthropods and nematodes. Mainly maternally inherited, these so-called sex parasites have selected several strategies that increase their vertical dispersion in host populations. However, the lack of congruence between the Wolbachia and their host phylogenies suggests frequent horizontal transfers. One way that could be used for horizontal Wolbachia transfers between individuals is predation. The aim of this study was to test whether horizontal passage of Wolbachia is possible when an uninfected terrestrial isopod eats an infected one. After having eaten Armadillidium vulgare harbouring Wolbachia, the predator-recipients (the two woodlice A. vulgare and Porcellio dilatatus dilatatus) that were initially Wolbachia-free were tested positive for the presence of Wolbachia both by quantitative PCR and Fluorescence in situ Hybridization (FISH). Even if the titers were low compared to vertically infected individuals, this constitutes the first demonstration of Wolbachia occurrence in various organs of an initially uninfected host after eating an infected one.}, } @article {pmid23593012, year = {2013}, author = {Ellegaard, KM and Klasson, L and Näslund, K and Bourtzis, K and Andersson, SG}, title = {Comparative genomics of Wolbachia and the bacterial species concept.}, journal = {PLoS genetics}, volume = {9}, number = {4}, pages = {e1003381}, pmid = {23593012}, issn = {1553-7404}, mesh = {Animals ; *Drosophila/genetics/microbiology ; *Genome, Bacterial ; Genomics ; New Caledonia ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Seychelles ; Species Specificity ; *Symbiosis/genetics/physiology ; *Wolbachia/genetics/physiology ; }, abstract = {The importance of host-specialization to speciation processes in obligate host-associated bacteria is well known, as is also the ability of recombination to generate cohesion in bacterial populations. However, whether divergent strains of highly recombining intracellular bacteria, such as Wolbachia, can maintain their genetic distinctness when infecting the same host is not known. We first developed a protocol for the genome sequencing of uncultivable endosymbionts. Using this method, we have sequenced the complete genomes of the Wolbachia strains wHa and wNo, which occur as natural double infections in Drosophila simulans populations on the Seychelles and in New Caledonia. Taxonomically, wHa belong to supergroup A and wNo to supergroup B. A comparative genomics study including additional strains supported the supergroup classification scheme and revealed 24 and 33 group-specific genes, putatively involved in host-adaptation processes. Recombination frequencies were high for strains of the same supergroup despite different host-preference patterns, leading to genomic cohesion. The inferred recombination fragments for strains of different supergroups were of short sizes, and the genomes of the co-infecting Wolbachia strains wHa and wNo were not more similar to each other and did not share more genes than other A- and B-group strains that infect different hosts. We conclude that Wolbachia strains of supergroup A and B represent genetically distinct clades, and that strains of different supergroups can co-exist in the same arthropod host without converging into the same species. This suggests that the supergroups are irreversibly separated and that barriers other than host-specialization are able to maintain distinct clades in recombining endosymbiont populations. Acquiring a good knowledge of the barriers to genetic exchange in Wolbachia will advance our understanding of how endosymbiont communities are constructed from vertically and horizontally transmitted genes.}, } @article {pmid23588850, year = {2013}, author = {Martínez-Rodríguez, P and Hernández-Pérez, M and Bella, JL}, title = {Detection of Spiroplasma and Wolbachia in the bacterial gonad community of Chorthippus parallelus.}, journal = {Microbial ecology}, volume = {66}, number = {1}, pages = {211-223}, pmid = {23588850}, issn = {1432-184X}, mesh = {Animals ; Female ; Gonads/microbiology ; Grasshoppers/genetics/*microbiology/*physiology ; Male ; Molecular Sequence Data ; Phylogeny ; Reproduction ; Spiroplasma/classification/genetics/*isolation & purification ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {We have recently detected the endosymbiont Wolbachia in multiple individuals and populations of the grasshopper Chorthippus parallelus (Orthoptera: acrididae). This bacterium induces reproductive anomalies, including cytoplasmic incompatibility. Such incompatibilities may help explain the maintenance of two distinct subspecies of this grasshopper, C. parallelus parallelus and C. parallelus erythropus, which are involved in a Pyrenean hybrid zone that has been extensively studied for the past 20 years, becoming a model system for the study of genetic divergence and speciation. To evaluate whether Wolbachia is the sole bacterial infection that might induce reproductive anomalies, the gonadal bacterial community of individuals from 13 distinct populations of C. parallelus was determined by denaturing gradient gel electrophoresis analysis of bacterial 16S rRNA gene fragments and sequencing. The study revealed low bacterial diversity in the gonads: a persistent bacterial trio consistent with Spiroplasma sp. and the two previously described supergroups of Wolbachia (B and F) dominated the gonad microbiota. A further evaluation of the composition of the gonad bacterial communities was carried out by whole cell hybridization. Our results confirm previous studies of the cytological distribution of Wolbachia in C. parallelus gonads and show a homogeneous infection by Spiroplasma. Spiroplasma and Wolbachia cooccurred in some individuals, but there was no significant association of Spiroplasma with a grasshopper's sex or with Wolbachia infection, although subtle trends might be detected with a larger sample size. This information, together with previous experimental crosses of this grasshopper, suggests that Spiroplasma is unlikely to contribute to sex-specific reproductive anomalies; instead, they implicate Wolbachia as the agent of the observed anomalies in C. parallelus.}, } @article {pmid23585900, year = {2013}, author = {Meriweather, M and Matthews, S and Rio, R and Baucom, RS}, title = {A 454 survey reveals the community composition and core microbiome of the common bed bug (Cimex lectularius) across an Urban Landscape.}, journal = {PloS one}, volume = {8}, number = {4}, pages = {e61465}, pmid = {23585900}, issn = {1932-6203}, mesh = {Animals ; Bacterial Typing Techniques ; Bedbugs/*microbiology ; Enterobacteriaceae/classification/*genetics/isolation & purification ; High-Throughput Nucleotide Sequencing ; Humans ; Metagenome/*genetics ; Ohio ; Principal Component Analysis ; RNA, Ribosomal, 16S/classification/*genetics/isolation & purification ; Urban Population ; Wolbachia/classification/*genetics/isolation & purification ; }, abstract = {Elucidating the spatial dynamic and core constituents of the microbial communities found in association with arthropod hosts is of crucial importance for insects that may vector human or agricultural pathogens. The hematophagous Cimex lectularius (Hemiptera: Cimicidae), known as the human bed bug, has made a recent resurgence in North America, as well as worldwide, potentially owing to increased travel, climate change and resistance to insecticides. A comprehensive survey of the bed bug microbiome has not been performed to date, nor has an assessment of the spatial dynamics of its microbiome. Here we present a survey of internal and external bed bug microbial communities by amplifying the V4-V6 hypervariable region of the 16S rDNA gene region followed by 454 Titanium sequencing using 31 individuals from eight distinct collection locations obtained from residences in Cincinnati, OH. Across all samples, 97% of the microbial community is made up of two dominant OTUs, previously identified as the α-proteobacterium Wolbachia and an unnamed γ-proteobacterium from the Enterobacteriaceae. Microbial communities varied among host locations for measures of community diversity and exhibited structure according to collection location. This broad survey represents the most in-depth assessment, to date, of the microbes that associate with bed bugs.}, } @article {pmid23584755, year = {2013}, author = {Schiefer, A and Vollmer, J and Lämmer, C and Specht, S and Lentz, C and Ruebsamen-Schaeff, H and Brötz-Oesterhelt, H and Hoerauf, A and Pfarr, K}, title = {The ClpP peptidase of Wolbachia endobacteria is a novel target for drug development against filarial infections.}, journal = {The Journal of antimicrobial chemotherapy}, volume = {68}, number = {8}, pages = {1790-1800}, doi = {10.1093/jac/dkt105}, pmid = {23584755}, issn = {1460-2091}, mesh = {Anti-Bacterial Agents/isolation & purification/*pharmacology ; Depsipeptides/isolation & purification/*pharmacology ; Endopeptidase Clp/*antagonists & inhibitors ; Filaricides/isolation & purification/*pharmacology ; Microscopy, Electron ; Microscopy, Fluorescence ; Protease Inhibitors/isolation & purification/*pharmacology ; Wolbachia/cytology/*drug effects/*enzymology/ultrastructure ; }, abstract = {BACKGROUND: Filarial infections causing lymphatic filariasis or onchocerciasis (river blindness) can be treated with antibiotics (e.g. doxycycline) targeting the essential endosymbiotic Wolbachia bacteria. The depletion of Wolbachia inhibits worm development and causes worm death. Available antibiotics have restrictions for use in children and pregnant or breastfeeding women. Therefore, alternative antibiotics are needed that can be given to all members of the population and that are active with a shorter therapy time. Antibiotics of the acyldepsipeptide class have been shown to inhibit the growth of bacteria by overactivating the peptidase ClpP. The novel mode of action of this class of antibiotics could lead to faster killing of intracellular bacteria.

OBJECTIVES: To characterize acyldepsipeptide activity against the Wolbachia ClpP.

METHODS: The activity of acyldepsipeptides was investigated against Wolbachia in vitro in insect cells and also against worms in culture. In addition, structural effects were investigated by fluorescence microscopy and electron microscopy. The activity of ClpP was also investigated in vitro.

RESULTS: We show that acyldepsipeptides are active against recombinant Wolbachia ClpP and endobacteria resident within insect cells in vitro, and some derivatives were also active against filarial worms in culture. As a consequence of treatment, the worms became immotile and died, the latter confirmed by a viability assay.

CONCLUSIONS: The mode of action of the acyldepsipeptides in Wolbachia is the dysregulation of ClpP, causing the uncontrolled degradation of proteins, including the cell division protein FtsZ. Our results demonstrate that wolbachial ClpP is a target for further antifilarial antibiotic discovery.}, } @article {pmid23576788, year = {2013}, author = {Hoffmann, AA and Turelli, M}, title = {Facilitating Wolbachia introductions into mosquito populations through insecticide-resistance selection.}, journal = {Proceedings. Biological sciences}, volume = {280}, number = {1760}, pages = {20130371}, pmid = {23576788}, issn = {1471-2954}, mesh = {Aedes/*genetics/*microbiology/virology ; Animals ; Australia ; *Biological Control Agents ; Dengue/*prevention & control ; Disease Resistance/genetics ; Female ; Genotype ; Humans ; Insect Vectors/*microbiology ; Male ; Models, Biological ; Mosquito Control/*methods ; Reproduction/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia infections are being introduced into mosquito vectors of human diseases following the discovery that they can block transmission of disease agents. This requires mosquitoes infected with the disease-blocking Wolbachia to successfully invade populations lacking the infection. While this process is facilitated by features of Wolbachia, particularly their ability to cause cytoplasmic incompatibility, blocking Wolbachia may produce deleterious effects, such as reduced host viability or fecundity, that inhibit successful local introductions and subsequent spatial spread. Here, we outline an approach to facilitate the introduction and spread of Wolbachia infections by coupling Wolbachia introduction to resistance to specific classes of insecticides. The approach takes advantage of very high maternal transmission fidelity of Wolbachia infections in mosquitoes, complete incompatibility between infected males and uninfected females, the widespread occurrence of insecticide resistance, and the widespread use of chemical control in disease-endemic countries. This approach is easily integrated into many existing control strategies, provides population suppression during release and might be used to introduce Wolbachia infections even with high and seasonally dependent deleterious effects, such as the wMelPop infection introduced into Aedes aegypti for dengue control. However, possible benefits will need to be weighed against concerns associated with the introduction of resistance alleles.}, } @article {pmid23556030, year = {2013}, author = {Blagrove, MS and Arias-Goeta, C and Di Genua, C and Failloux, AB and Sinkins, SP}, title = {A Wolbachia wMel transinfection in Aedes albopictus is not detrimental to host fitness and inhibits Chikungunya virus.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {3}, pages = {e2152}, pmid = {23556030}, issn = {1935-2735}, support = {//Wellcome Trust/United Kingdom ; 095121/WT_/Wellcome Trust/United Kingdom ; BB/G016976/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Chikungunya virus/*growth & development ; Drosophila melanogaster/microbiology ; Female ; Male ; *Microbial Interactions ; Saliva/virology ; Wolbachia/*growth & development/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia inherited intracellular bacteria can manipulate the reproduction of their insect hosts through cytoplasmic incompatibility (CI), and certain strains have also been shown to inhibit the replication or dissemination of viruses. Wolbachia strains also vary in their relative fitness effects on their hosts and this is a particularly important consideration with respect to the potential of newly created transinfections for use in disease control.

In Aedes albopictus mosquitoes transinfected with the wMel strain from Drosophila melanogaster, which we previously reported to be unable to transmit dengue in lab challenges, no significant detrimental effects were observed on egg hatch rate, fecundity, adult longevity or male mating competitiveness. All these parameters influence the population dynamics of Wolbachia, and the data presented are favourable with respect to the aim of taking wMel to high population frequency. Challenge with the chikungunya (CHIKV) virus, for which Ae. albopictus is an important vector, was conducted and the presence of wMel abolished CHIKV dissemination to the saliva.

CONCLUSIONS/SIGNIFICANCE: Taken together, these data suggest that introducing wMel into natural Ae. albopictus populations using bidirectional CI could be an efficient strategy for preventing or reducing the transmission of arboviruses by this species.}, } @article {pmid23555728, year = {2013}, author = {Baton, LA and Pacidônio, EC and Gonçalves, DS and Moreira, LA}, title = {wFlu: characterization and evaluation of a native Wolbachia from the mosquito Aedes fluviatilis as a potential vector control agent.}, journal = {PloS one}, volume = {8}, number = {3}, pages = {e59619}, pmid = {23555728}, issn = {1932-6203}, mesh = {Aedes/cytology/drug effects/growth & development/*microbiology ; Animals ; Cytoplasm/microbiology ; Female ; Fertility ; Humans ; Insect Control/*methods ; Insect Vectors/cytology/drug effects/growth & development/*microbiology ; Male ; Oocysts/microbiology ; Organ Specificity ; Pest Control, Biological/*methods ; Sex Characteristics ; Tetracycline/pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {There is currently considerable interest and practical progress in using the endosymbiotic bacteria Wolbachia as a vector control agent for human vector-borne diseases. Such vector control strategies may require the introduction of multiple, different Wolbachia strains into target vector populations, necessitating the identification and characterization of appropriate endosymbiont variants. Here, we report preliminary characterization of wFlu, a native Wolbachia from the neotropical mosquito Aedes fluviatilis, and evaluate its potential as a vector control agent by confirming its ability to cause cytoplasmic incompatibility, and measuring its effect on three parameters determining host fitness (survival, fecundity and fertility), as well as vector competence (susceptibility) for pathogen infection. Using an aposymbiotic strain of Ae. fluviatilis cured of its native Wolbachia by antibiotic treatment, we show that in its natural host wFlu causes incomplete, but high levels of, unidirectional cytoplasmic incompatibility, has high rates of maternal transmission, and no detectable fitness costs, indicating a high capacity to rapidly spread through host populations. However, wFlu does not inhibit, and even enhances, oocyst infection with the avian malaria parasite Plasmodium gallinaceum. The stage- and sex-specific density of wFlu was relatively low, and with limited tissue distribution, consistent with the lack of virulence and pathogen interference/symbiont-mediated protection observed. Unexpectedly, the density of wFlu was also shown to be specifically-reduced in the ovaries after bloodfeeding Ae. fluviatilis. Overall, our observations indicate that the Wolbachia strain wFlu has the potential to be used as a vector control agent, and suggests that appreciable mutualistic coevolution has occurred between this endosymbiont and its natural host. Future work will be needed to determine whether wFlu has virulent host effects and/or exhibits pathogen interference when artificially-transfected to the novel mosquito hosts that are the vectors of human pathogens.}, } @article {pmid23550173, year = {2012}, author = {Kraaijeveld, K and Bast, J}, title = {Transposable element proliferation as a possible side effect of endosymbiont manipulations.}, journal = {Mobile genetic elements}, volume = {2}, number = {5}, pages = {253-256}, pmid = {23550173}, issn = {2159-2543}, abstract = {The mode of reproduction has been predicted to affect the proliferation of transposable elements (TEs). A population that switches from sexual to asexual reproduction could either accumulate TEs because purifying selection becomes less efficient, or a decrease in TE load because the opportunity for horizontal transmission is reduced. A third possibility is that the mechanism that induces asexual reproduction affects TE dynamics as a side effect. We propose two such mechanisms that might explain recently described patterns of TE abundance in sexual and asexual lineages of the parasitoid wasp Leptopilina clavipes. Asexual reproduction in this species is induced by endosymbiotic Wolbachia bacteria. In order to achieve parthenogenesis in its host, Wolbachia might remove methylation or interfere with Argonaute proteins. Both methylation and Argonaute proteins are known to control TE activity in other species. By interfering with either, Wolbachia might therefore secondarily hamper the control of specific TEs.}, } @article {pmid23540123, year = {2013}, author = {Jacups, SP and Ball, TS and Paton, CJ and Johnson, PH and Ritchie, SA}, title = {Operational use of household bleach to "crash and release" Aedes aegypti prior to Wolbachia-infected mosquito release.}, journal = {Journal of medical entomology}, volume = {50}, number = {2}, pages = {344-351}, doi = {10.1603/me12043}, pmid = {23540123}, issn = {0022-2585}, mesh = {Acanthaceae/drug effects ; Aedes/*drug effects/microbiology ; Age Factors ; Animals ; Araceae/drug effects ; Fresh Water/chemistry ; Insect Repellents/analysis/*pharmacology/toxicity ; Insecticides/analysis/*pharmacology/toxicity ; Larva/drug effects/microbiology ; Mosquito Control/*methods ; Oviposition/drug effects ; Ovum/drug effects/microbiology ; Pupa/drug effects/microbiology ; Sodium Hypochlorite/analysis/*pharmacology/toxicity ; Time Factors ; Wolbachia/physiology ; }, abstract = {Dengue (family Flaviviridae, genus Flavivirus, DENV) remains the leading arboviral cause of mortality in the tropics. Wolbachia pipientis has been shown to interrupt DENV transmission and is presently being trialled as a biological control. However, deployment issues have arisen on methods to temporarily suppress wild mosquito populations before Wolbachia-infected mosquito releases. By suppressing wild populations, fewer Ae. aegypti releases are required to achieve a sustainable Wolbachia density threshold. Furthermore, public distress is reduced. This study tests the application of domestic bleach (4% NaCIO) to temporarily "crash" immature Aedes populations in water-filled containers. Spray application NaClO (215 ppm) resulted in a mean 48-h mortality of 100, 100, 97, and 88% of eggs, second-instar larvae, fourth-instar larvae, and pupae, respectively. In the field, NaClO delayed ovipositing by 9 d in cooler months, and 11 d in hotter months, after which oviposition resumed in treated receptacles. We found bleach treatment of pot-plant bases did not cause wilting, yellowing, or dropping of leaves in two ornamental plants species. Domestically available NaClO could be adopted for a "crash and release" strategy to temporarily suppress wild populations of Ae. aegypti in containers before release of Wolbachia-infected mosquitoes. The "crash and release" strategy is also applicable to other mosquito species, e.g., Aedes albopictus (Skuse), in strategies using released mosquitoes.}, } @article {pmid23535905, year = {2013}, author = {Hughes, H and Britton, NF}, title = {Modelling the use of Wolbachia to control dengue fever transmission.}, journal = {Bulletin of mathematical biology}, volume = {75}, number = {5}, pages = {796-818}, doi = {10.1007/s11538-013-9835-4}, pmid = {23535905}, issn = {1522-9602}, mesh = {Aedes/microbiology/virology ; Animals ; Dengue/*prevention & control/transmission ; Dengue Virus/pathogenicity ; Female ; Humans ; Insect Vectors/microbiology/virology ; Male ; Mathematical Concepts ; *Models, Biological ; Pest Control, Biological/methods/statistics & numerical data ; Wolbachia/*physiology ; }, abstract = {Experiments and field trials have shown that the intracellular bacterium Wolbachia may be introduced into populations of the mosquito Aedes aegypti, the primary vector for dengue fever. In the absence of Wolbachia, a mosquito acquiring the dengue virus from an infected human enters an exposed (infected but not infectious) period before becoming infectious itself. A Wolbachia-infected mosquito that acquires dengue (i) may have a reduced lifespan, so that it is less likely to survive the exposed period and become infectious, and (ii) may have a reduced ability to transmit dengue, even if it has survived the exposed period. Wolbachia introduction has therefore been suggested as a potential dengue control measure. We set up a mathematical model for the system to investigate this suggestion and to evaluate the desirable properties of the Wolbachia strain to be introduced. We show that Wolbachia has excellent potential for dengue control in areas where R 0 is not too large. However, if R 0 is large, Wolbachia strains that reduce but do not eliminate dengue transmission have little effect on endemic steady states or epidemic sizes. Unless control measures to reduce R 0 by reducing mosquito populations are also put in place, it may be worth the extra effort in such cases to introduce Wolbachia strains that eliminate dengue transmission completely.}, } @article {pmid23525074, year = {2013}, author = {Desjardins, CA and Cerqueira, GC and Goldberg, JM and Dunning Hotopp, JC and Haas, BJ and Zucker, J and Ribeiro, JM and Saif, S and Levin, JZ and Fan, L and Zeng, Q and Russ, C and Wortman, JR and Fink, DL and Birren, BW and Nutman, TB}, title = {Genomics of Loa loa, a Wolbachia-free filarial parasite of humans.}, journal = {Nature genetics}, volume = {45}, number = {5}, pages = {495-500}, pmid = {23525074}, issn = {1546-1718}, support = {DP2 OD007372/OD/NIH HHS/United States ; HHSN272200900018C/AI/NIAID NIH HHS/United States ; Z01 AI000512-21//Intramural NIH HHS/United States ; 1-DP2-OD007372/OD/NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/genetics ; Filariasis/*genetics/parasitology ; Filarioidea/*genetics/parasitology ; Genes, Helminth/*genetics ; *Genome, Helminth ; Humans ; Loa/*genetics ; Molecular Sequence Data ; Phylogeny ; Protein Kinases/*metabolism ; Symbiosis ; Wolbachia/*genetics ; Wuchereria bancrofti/genetics ; }, abstract = {Loa loa, the African eyeworm, is a major filarial pathogen of humans. Unlike most filariae, L. loa does not contain the obligate intracellular Wolbachia endosymbiont. We describe the 91.4-Mb genome of L. loa and that of the related filarial parasite Wuchereria bancrofti and predict 14,907 L. loa genes on the basis of microfilarial RNA sequencing. By comparing these genomes to that of another filarial parasite, Brugia malayi, and to those of several other nematodes, we demonstrate synteny among filariae but not with nonparasitic nematodes. The L. loa genome encodes many immunologically relevant genes, as well as protein kinases targeted by drugs currently approved for use in humans. Despite lacking Wolbachia, L. loa shows no new metabolic synthesis or transport capabilities compared to other filariae. These results suggest that the role of Wolbachia in filarial biology is more subtle than previously thought and reveal marked differences between parasitic and nonparasitic nematodes.}, } @article {pmid23517690, year = {2013}, author = {Vollmer, J and Schiefer, A and Schneider, T and Jülicher, K and Johnston, KL and Taylor, MJ and Sahl, HG and Hoerauf, A and Pfarr, K}, title = {Requirement of lipid II biosynthesis for cell division in cell wall-less Wolbachia, endobacteria of arthropods and filarial nematodes.}, journal = {International journal of medical microbiology : IJMM}, volume = {303}, number = {3}, pages = {140-149}, doi = {10.1016/j.ijmm.2013.01.002}, pmid = {23517690}, issn = {1618-0607}, mesh = {Animals ; Arthropods/microbiology ; Biosynthetic Pathways/genetics ; *Cell Division ; Cell Wall/*metabolism ; Gene Expression Profiling ; Nematoda/microbiology ; Peptidoglycan/biosynthesis ; Uridine Diphosphate N-Acetylmuramic Acid/*analogs & derivatives/biosynthesis ; Wolbachia/*physiology ; }, abstract = {Obligate Wolbachia endobacteria have a reduced genome and retained genes are hypothesized to be crucial for survival. Although intracellular bacteria do not need a stress-bearing peptidoglycan cell wall, Wolbachia encode proteins necessary to synthesize the peptidoglycan precursor lipid II. The activity of the enzymes catalyzing the last two steps of this pathway was previously shown, and Wolbachia are sensitive to inhibition of lipid II synthesis. A puzzling characteristic of Wolbachia is the lack of genes for l-amino acid racemases essential for lipid II synthesis. Transcription analysis showed the expression of a possible alternative racemase metC, and recombinant Wolbachia MetC indeed had racemase activity that may substitute for the absent l-Ala racemase. However, enzymes needed to form mature peptidoglycan are absent and the function of Wolbachia lipid II is unknown. Inhibition of lipid II biosynthesis resulted in enlargement of Wolbachia cells and redistribution of Wolbachia peptidoglycan-associated lipoprotein, demonstrating that lipid II is required for coordinated cell division and may interact with the lipoprotein. We conclude that lipid II is essential for Wolbachia cell division and that this function is potentially conserved in the Gram-negative bacteria.}, } @article {pmid23517577, year = {2013}, author = {Cockburn, SN and Haselkorn, TS and Hamilton, PT and Landzberg, E and Jaenike, J and Perlman, SJ}, title = {Dynamics of the continent-wide spread of a Drosophila defensive symbiont.}, journal = {Ecology letters}, volume = {16}, number = {5}, pages = {609-616}, doi = {10.1111/ele.12087}, pmid = {23517577}, issn = {1461-0248}, mesh = {Adaptation, Physiological ; Animals ; Canada ; DNA, Mitochondrial ; Drosophila/genetics/*microbiology/*parasitology ; Electron Transport Complex IV/genetics ; Female ; Fertility ; Genetic Variation ; Selection, Genetic ; Spiroplasma/pathogenicity/*physiology ; Symbiosis ; Tylenchida/pathogenicity ; Wolbachia ; }, abstract = {Facultative symbionts can represent important sources of adaptation for their insect hosts and thus have the potential for rapid spread. Drosophila neotestacea harbours a heritable symbiont, Spiroplasma, that confers protection against parasitic nematodes. We previously found a cline in Spiroplasma prevalence across central Canada, ending abruptly at the Rocky Mountains. Resampling these populations 9 years later revealed that Spiroplasma had increased substantially across the region, resembling a Fisherian wave of advance. Associations between Spiroplasma infection and host mitochondrial DNA indicate that the increase was due to local increase of Spiroplasma-infected flies. Finally, we detected Spiroplasma west of the Rocky Mountains for the first time and showed that defence against nematodes occurs in flies with a western genetic background. Because nematode infection is common throughout D. neotestacea's range, we expect Spiroplasma to spread to the Pacific coast.}, } @article {pmid23508299, year = {2013}, author = {Eleftherianos, I and Atri, J and Accetta, J and Castillo, JC}, title = {Endosymbiotic bacteria in insects: guardians of the immune system?.}, journal = {Frontiers in physiology}, volume = {4}, number = {}, pages = {46}, pmid = {23508299}, issn = {1664-042X}, abstract = {Insects have evolved obligate, mutualistic interactions with bacteria without further transmission to other eukaryotic organisms. Such long-term obligate partnerships between insects and bacteria have a profound effect on various physiological functions of the host. Here we provide an overview of the effects of endosymbiotic bacteria on the insect immune system as well as on the immune response of insects to pathogenic infections. Potential mechanisms through which endosymbionts can affect the ability of their host to resist an infection are discussed in the light of recent findings. We finally point out unresolved questions for future research and speculate how the current knowledge can be employed to design and implement measures for the effective control of agricultural insect pests and vectors of diseases.}, } @article {pmid23505512, year = {2013}, author = {Rey, O and Estoup, A and Facon, B and Loiseau, A and Aebi, A and Duron, O and Vavre, F and Foucaud, J}, title = {Distribution of endosymbiotic reproductive manipulators reflects invasion process and not reproductive system polymorphism in the little fire ant Wasmannia auropunctata.}, journal = {PloS one}, volume = {8}, number = {3}, pages = {e58467}, pmid = {23505512}, issn = {1932-6203}, mesh = {Animals ; Ants/*microbiology/*physiology ; Female ; Genes, Bacterial ; Genotype ; Male ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny ; *Reproduction ; *Symbiosis ; Wolbachia/classification/*genetics ; }, abstract = {Endosymbiotic reproductive manipulators may have drastic effects on the ecological and evolutionary dynamics of their hosts. The prevalence of these endosymbionts reflects both their ability to manipulate their hosts and the history of the host populations. The little fire ant Wasmannia auropunctata displays a polymorphism in both its reproductive system (sexual versus clonal populations) and the invasive status of its populations (associated to a habitat shift). We first screened for the presence of a diverse array of reproductive parasites in sexual and clonal populations of W. auropunctata, as a means to investigate the role of endosymbionts in reproductive phenotypes. Wolbachia was the only symbiont found and we then focused on its worldwide distribution and diversity in natural populations of W. auropunctata. Using a multilocus scheme, we further characterized the Wolbachia strains present in these populations. We found that almost all the native sexual populations and only a few clonal populations are infected by Wolbachia. The presence of similar Wolbachia strains in both sexual and clonal populations indicates that they are probably not the cause of the reproductive system polymorphism. The observed pattern seems rather associated to the invasion process of W. auropunctata. In particular, the observed loss of Wolbachia in clonal populations, that recurrently emerged from sexual populations, likely resulted from natural heat treatment and/or relaxed selection during the shift in habitat associated to the invasion process.}, } @article {pmid23490256, year = {2013}, author = {Albertson, R and Tan, V and Leads, RR and Reyes, M and Sullivan, W and Casper-Lindley, C}, title = {Mapping Wolbachia distributions in the adult Drosophila brain.}, journal = {Cellular microbiology}, volume = {15}, number = {9}, pages = {1527-1544}, doi = {10.1111/cmi.12136}, pmid = {23490256}, issn = {1462-5822}, mesh = {Animals ; Bacterial Load ; Brain/microbiology ; Drosophila/*microbiology ; Fat Body/microbiology ; Wolbachia/*isolation & purification ; }, abstract = {The maternally inherited bacterium Wolbachia infects the germline of most arthropod species. Using Drosophila simulans and D. melanogaster, we demonstrate that localization of Wolbachia to the fat bodies and adult brain is likely also a conserved feature of Wolbachia infection. Examination of three Wolbachia strains (WMel , WRiv , WPop) revealed that the bacteria preferentially concentrate in the central brain with low titres in the optic lobes. Distribution within regions of the central brain is largely determined by the Wolbachia strain, while the titre is influenced by both, the host species and the bacteria strain. In neurons of the central brain and ventral nerve cord, Wolbachia preferentially localizes to the neuronal cell bodies but not to axons. All examined Wolbachia strains are present intracellularly or in extracellular clusters, with the pathogenic WPop strain exhibiting the largest and most abundant clusters. We also discovered that 16 of 40 lines from the Drosophila Genetic Reference Panel are Wolbachia infected. Direct comparison of Wolbachia infected and cured lines from this panel reveals that differences in physiological traits (chill coma recovery, starvation, longevity) are partially due to host line influences. In addition, a tetracycline-induced increase in Drosophila longevity was detected many generations after treatment.}, } @article {pmid23482460, year = {2012}, author = {Geniez, S and Foster, JM and Kumar, S and Moumen, B and Leproust, E and Hardy, O and Guadalupe, M and Thomas, SJ and Boone, B and Hendrickson, C and Bouchon, D and Grève, P and Slatko, BE}, title = {Targeted genome enrichment for efficient purification of endosymbiont DNA from host DNA.}, journal = {Symbiosis (Philadelphia, Pa.)}, volume = {58}, number = {1-3}, pages = {201-207}, pmid = {23482460}, issn = {0334-5114}, abstract = {Wolbachia endosymbionts are widespread in arthropods and are generally considered reproductive parasites, inducing various phenotypes including cytoplasmic incompatibility, parthenogenesis, feminization and male killing, which serve to promote their spread through populations. In contrast, Wolbachia infecting filarial nematodes that cause human diseases, including elephantiasis and river blindness, are obligate mutualists. DNA purification methods for efficient genomic sequencing of these unculturable bacteria have proven difficult using a variety of techniques. To efficiently capture endosymbiont DNA for studies that examine the biology of symbiosis, we devised a parallel strategy to an earlier array-based method by creating a set of SureSelect™ (Agilent) 120-mer target enrichment RNA oligonucleotides ("baits") for solution hybrid selection. These were designed from Wolbachia complete and partial genome sequences in GenBank and were tiled across each genomic sequence with 60 bp overlap. Baits were filtered for homology against host genomes containing Wolbachia using BLAT and sequences with significant host homology were removed from the bait pool. Filarial parasite Brugia malayi DNA was used as a test case, as the complete sequence of both Wolbachia and its host are known. DNA eluted from capture was size selected and sequencing samples were prepared using the NEBNext® Sample Preparation Kit. One-third of a 50 nt paired-end sequencing lane on the HiSeq™ 2000 (Illumina) yielded 53 million reads and the entirety of the Wolbachia genome was captured. We then used the baits to isolate more than 97.1 % of the genome of a distantly related Wolbachia strain from the crustacean Armadillidium vulgare, demonstrating that the method can be used to enrich target DNA from unculturable microbes over large evolutionary distances.}, } @article {pmid23472225, year = {2013}, author = {Siozios, S and Cestaro, A and Kaur, R and Pertot, I and Rota-Stabelli, O and Anfora, G}, title = {Draft Genome Sequence of the Wolbachia Endosymbiont of Drosophila suzukii.}, journal = {Genome announcements}, volume = {1}, number = {1}, pages = {}, pmid = {23472225}, issn = {2169-8287}, support = {MR/K001744/1/MRC_/Medical Research Council/United Kingdom ; }, abstract = {Wolbachia is one of the most successful and abundant symbiotic bacteria in nature, infecting more than 40% of the terrestrial arthropod species. Here we report the draft genome sequence of a novel Wolbachia strain named "wSuzi" that was retrieved from the genome sequencing of its host, the invasive pest Drosophila suzukii.}, } @article {pmid23460918, year = {2013}, author = {Hall-Mendelin, S and Allcock, R and Kresoje, N and van den Hurk, AF and Warrilow, D}, title = {Detection of arboviruses and other micro-organisms in experimentally infected mosquitoes using massively parallel sequencing.}, journal = {PloS one}, volume = {8}, number = {2}, pages = {e58026}, pmid = {23460918}, issn = {1932-6203}, mesh = {Aedes/*microbiology/*virology ; Animals ; Arbovirus Infections/genetics/*microbiology/*virology ; Arboviruses/*genetics/*isolation & purification ; Base Sequence ; Genome, Viral/genetics ; High-Throughput Nucleotide Sequencing/*methods ; Humans ; RNA, Ribosomal, 16S/genetics ; Sheep ; Wolbachia/genetics ; }, abstract = {Human disease incidence attributed to arbovirus infection is increasing throughout the world, with effective control interventions limited by issues of sustainability, insecticide resistance and the lack of effective vaccines. Several promising control strategies are currently under development, such as the release of mosquitoes trans-infected with virus-blocking Wolbachia bacteria. Implementation of any control program is dependent on effective virus surveillance and a thorough understanding of virus-vector interactions. Massively parallel sequencing has enormous potential for providing comprehensive genomic information that can be used to assess many aspects of arbovirus ecology, as well as to evaluate novel control strategies. To demonstrate proof-of-principle, we analyzed Aedes aegypti or Aedes albopictus experimentally infected with dengue, yellow fever or chikungunya viruses. Random amplification was used to prepare sufficient template for sequencing on the Personal Genome Machine. Viral sequences were present in all infected mosquitoes. In addition, in most cases, we were also able to identify the mosquito species and mosquito micro-organisms, including the bacterial endosymbiont Wolbachia. Importantly, naturally occurring Wolbachia strains could be differentiated from strains that had been trans-infected into the mosquito. The method allowed us to assemble near full-length viral genomes and detect other micro-organisms without prior sequence knowledge, in a single reaction. This is a step toward the application of massively parallel sequencing as an arbovirus surveillance tool. It has the potential to provide insight into virus transmission dynamics, and has applicability to the post-release monitoring of Wolbachia in mosquito populations.}, } @article {pmid23440444, year = {2013}, author = {Robert, CA and Frank, DL and Leach, KA and Turlings, TC and Hibbard, BE and Erb, M}, title = {Direct and indirect plant defenses are not suppressed by endosymbionts of a specialist root herbivore.}, journal = {Journal of chemical ecology}, volume = {39}, number = {4}, pages = {507-515}, pmid = {23440444}, issn = {1573-1561}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Coleoptera/drug effects/growth & development/*physiology ; Gene Expression Regulation, Plant ; Herbivory/drug effects ; Larva/physiology ; Nematoda/drug effects/physiology ; Plant Roots/chemistry/metabolism/microbiology ; Polycyclic Sesquiterpenes ; Sesquiterpenes/chemistry/pharmacology ; Symbiosis/drug effects ; Volatile Organic Compounds/chemistry/metabolism ; Wolbachia/drug effects/isolation & purification ; Zea mays/chemistry/*metabolism/microbiology ; }, abstract = {Insect endosymbionts influence many important metabolic and developmental processes of their host. It has been speculated that they may also help to manipulate and suppress plant defenses to the benefit of herbivores. Recently, endosymbionts of the root herbivore Diabrotica virgifera virgifera have been reported to suppress the induction of defensive transcripts in maize roots, which may explain the finding of another study that once attacked plants become more susceptible to subsequent D. v. virgifera attack. To test this hypothesis, we cured D. v. virgifera from its major endosymbiont Wolbachia and tested whether endosymbiont-free individuals elicit different defense responses in maize roots. The presence of Wolbachia did not alter the induction of defense marker genes and resistance in a susceptible maize hybrid and a resistant line. Furthermore, attacked maize plants emitted the same amount of (E)-β-caryophyllene, a volatile signal that serves as foraging cue for both entomopathogenic nematodes and D. v. virgifera. Finally, the effectiveness of the entomopathogenic nematode Heterorhabditis bacteriophora to infest D. v. virgifera was not changed by curing the larvae from their endosymbionts. These results show that the defense mechanisms of maize are not affected by Wolbachia. Consequently, D. v. virgifera does not seem to derive any plant-defense mediated benefits from its major endosymbiont.}, } @article {pmid23438747, year = {2013}, author = {Lentz, CS and Halls, V and Hannam, JS and Niebel, B and Strübing, U and Mayer, G and Hoerauf, A and Famulok, M and Pfarr, KM}, title = {A selective inhibitor of heme biosynthesis in endosymbiotic bacteria elicits antifilarial activity in vitro.}, journal = {Chemistry & biology}, volume = {20}, number = {2}, pages = {177-187}, doi = {10.1016/j.chembiol.2012.11.009}, pmid = {23438747}, issn = {1879-1301}, mesh = {Animals ; Antiprotozoal Agents/chemistry/*pharmacology/therapeutic use ; Benzimidazoles/chemistry/*pharmacology/therapeutic use ; Drug Design ; Elephantiasis, Filarial/drug therapy ; Filarioidea/*drug effects ; Heme/*biosynthesis ; High-Throughput Screening Assays ; Humans ; Kinetics ; Magnesium/chemistry/metabolism ; Porphobilinogen Synthase/antagonists & inhibitors/metabolism ; Symbiosis ; Thiophenes/chemistry/*pharmacology/therapeutic use ; Wolbachia/enzymology/*metabolism ; }, abstract = {Lymphatic filariasis and onchocerciasis are severe diseases caused by filarial worms and affect more than 150 million people worldwide. Endosymbiotic α-proteobacteria Wolbachia are essential for these parasites throughout their life cycle. Using a high-throughput chemical screen, we identified a benzimidazole compound, wALADin1, that selectively targets the δ-aminolevulinic acid dehydratase (ALAD) of Wolbachia (wALAD) and exhibits macrofilaricidal effects on Wolbachia-containing filarial worms in vitro. wALADin1 is a mixed competitive/noncompetitive inhibitor that interferes with the Mg(2+)-induced activation of wALAD. This mechanism inherently excludes activity against the Zn(2+)-dependent human ortholog and might be translatable to Mg(2+)-responsive orthologs of other bacterial or protozoan pathogens. The specificity profile of wALADin1 derivatives reveals chemical features responsible for inhibitory potency and species selectivity. Our findings validate wALADins as a basis for developing potent leads that meet current requirements for antifilarial drugs.}, } @article {pmid23433646, year = {2013}, author = {García-Guasch, L and Caro-Vadillo, A and Manubens-Grau, J and Carretón, E and Morchón, R and Simón, F and Kramer, LH and Montoya-Alonso, JA}, title = {Is Wolbachia participating in the bronchial reactivity of cats with heartworm associated respiratory disease?.}, journal = {Veterinary parasitology}, volume = {196}, number = {1-2}, pages = {130-135}, doi = {10.1016/j.vetpar.2013.01.060}, pmid = {23433646}, issn = {1873-2550}, mesh = {Animals ; Antibodies, Bacterial/blood ; Bronchoconstriction ; Cat Diseases/microbiology/parasitology/*pathology ; Cats ; Dirofilariasis/*microbiology/pathology ; Female ; Immunoglobulin G/blood ; Male ; Plethysmography ; Respiratory Tract Diseases/etiology/pathology/*veterinary ; Serologic Tests ; Wolbachia/*physiology ; }, abstract = {Heartworm associated respiratory disease is a pulmonary syndrome in cats that results from the vascular and parenchymal inflammatory response associated with the arrival and death of Dirofilaria immitis worms into the distal pulmonary arteries. This parasite harbors intracellular Wolbachia, an endosymbiont bacteria. The association between the parasite and the bacteria is obligatory. Some studies suggest the involvement of Wolbachia in the development of the inflammatory reaction and in the polarization of the host immune response against the parasite. Barometric whole-body plethysmography is a non-invasive pulmonary function test that allows a dynamic study of breathing patterns and is useful to study airway disease and the response to different treatments. The aim of this prospective non-blinded study was to compare the influence of Wolbachia upon the respiratory function variables in a population of cats seropositive to D. immitis by use of Barometric whole-body plethysmography. Fourteen seropositive cats to Wolbachia and eight seronegative cats were put into the plethysmograph chamber and different respiratory variables were measured. The results were analyzed and compared between the two groups of animals. Significant differences were found for bronchoconstriction index variables PAU (pause) (P-value<0.05) and Penh (enhanced pause) (P-value<0.05). The results obtained in our study suggest that Wolbachia seems to produce a greater acute inflammatory response at bronchial, vascular and parenchymal level worsening the state of broncho-reactivity associated with the presence of seropositivity to D. immitis in cats.}, } @article {pmid23429887, year = {2013}, author = {Zhang, YK and Zhang, KJ and Sun, JT and Yang, XM and Ge, C and Hong, XY}, title = {Diversity of Wolbachia in natural populations of spider mites (genus Tetranychus): evidence for complex infection history and disequilibrium distribution.}, journal = {Microbial ecology}, volume = {65}, number = {3}, pages = {731-739}, pmid = {23429887}, issn = {1432-184X}, mesh = {Animals ; *Biodiversity ; China ; Endophytes/classification/genetics/isolation & purification/*physiology ; Female ; Host Specificity ; Male ; Molecular Sequence Data ; Phylogeny ; Tetranychidae/classification/genetics/*microbiology ; Wolbachia/classification/genetics/isolation & purification/*physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria that commonly infect arthropods and cause reproductive disorders in host. Within several Tetranychus species, Wolbachia have been detected and shown to affect their reproduction. However, little is known about their transmission and distribution patterns in natural populations of Tetranychus species. Here, we used multilocus sequence typing to confirm Wolbachia infection status and examined the relationship between Wolbachia infection status and host phylogeny, mitochondrial diversity, and geographical range in five Tetranychus species (Tetranychus truncatus, Tetranychus urticae, Tetranychus pueraricola, Tetranychus phaselus, and Tetranychus kanzawai) from 21 populations in China. The prevalence of Wolbachia within the five Tetranychus species ranged from 31.4 to 100 %, and the strains were remarkably diverse. Together, these observations indicate that Wolbachia was introduced to these populations on multiple separate occasions. As in other arthropods, the same Tetranychus species can accommodate very different strains, and identical Wolbachia occasionally infect different species. These observations suggest that Wolbachia are transmitted both vertically and horizontally. Horizontally, transmission is probably mediated by the host plants. The distribution patterns of Wolbachia were quite different among populations of the same species, suggesting that the dynamics of Wolbachia in nature may be affected by ecological and other factors.}, } @article {pmid23428527, year = {2012}, author = {Muniaraj, M and Paramasivan, R and Sunish, IP and Arunachalam, N and Mariappan, T and Jerald Leo, SV and Dhananjeyan, KJ}, title = {Detection of Wolbachia endobacteria in Culex quinquefasciatus by Gimenez staining and confirmation by PCR.}, journal = {Journal of vector borne diseases}, volume = {49}, number = {4}, pages = {258-261}, pmid = {23428527}, issn = {0972-9062}, mesh = {Animals ; Culex/classification/*microbiology ; DNA, Bacterial/analysis/genetics ; Elephantiasis, Filarial/transmission ; Female ; Insect Vectors/microbiology ; Ovary/microbiology ; Polymerase Chain Reaction/*methods ; Staining and Labeling/*methods ; Wolbachia/*genetics/*isolation & purification ; }, abstract = {BACKGROUND & OBJECTIVES: Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-egg incompatibility. Since they can also move horizontally across species boundaries, Wolbachia is gaining importance in recent days as it could be used as a biological control agent to control vector mosquitoes or for paratransgenic approaches. However, the study of Wolbachia requires sophisticated techniques such as PCR and cell culture facilities which cannot be affordable for many laboratories where the diseases transmitted by arthropod vectors are common. Hence, it would be beneficial to develop a simple method to detect the presence of Wolbachia in arthropods.

METHOD: In this study, we described a method of staining Wolbachia endobacteria, present in the reproductive tissues of mosquitoes. The reliability of this method was compared with Gram staining and PCR based detection.

RESULTS: The microscopic observation of the Gimenez stained smear prepared from the teased ovary of wild caught and Wolbachia (+) Cx. quinquefasciatus revealed the presence of pink coloured pleomorphic cells of Wolbachia ranging from cocci, comma shaped cells to bacillus and chain forms. The ovaries of Wolbachia (-) cured mosquito did not show any cell. Although Gram's staining is a reliable differential staining for the other bacteria, the bacterial cells in the smears from the ovaries of wild caught mosquitoes did not take the stain properly and the cells were not clearly visible. The PCR amplified product from the pooled remains of wild caught and Wolbachia (+) Cx. quinquefasciatus showed clear banding, whereas, no banding was observed for the negative control (distilled water) and Wolbachia (-) Cx. quinquefasciatus.

The Gimenez staining technique applied, could be used to detect the members of the endobacteria Wolbachia easily, even in a simple laboratory without any special facilities or even in the field condition and for handling large number of samples in a shorter duration.}, } @article {pmid23427945, year = {2013}, author = {Ali, M and Afzal, M and Bhattacharya, SM and Ahmad, FJ and Dinda, AK}, title = {Nanopharmaceuticals to target antifilarials: a comprehensive review.}, journal = {Expert opinion on drug delivery}, volume = {10}, number = {5}, pages = {665-678}, doi = {10.1517/17425247.2013.771630}, pmid = {23427945}, issn = {1744-7593}, mesh = {Animals ; Chemistry, Pharmaceutical ; *Drug Delivery Systems ; Elephantiasis, Filarial/*drug therapy ; Filaricides/*administration & dosage ; Filarioidea/drug effects ; Humans ; Liposomes/*chemistry ; Nanoparticles/*chemistry ; Wolbachia/drug effects ; }, abstract = {INTRODUCTION: Currently emphasized conventional chemotherapies for the elimination of lymphatic filariasis (LF) are imperfect due to unfocused targeting of poorly water-soluble antifilarial drugs. The profound location of drug targets (filarial parasites or wolbachia) within the complex anatomy of lymphatic tissues often necessitates prolonged treatment schedules with high doses leading to undesired side effects and poor patient compliance. Therefore, we need to reformulate antifilarial drugs taking the advantages of nanotechnology through a wide range of nanomedical carriers, which improve drug efficacy, increase bioavailability, and diminish toxicity.

AREAS COVERED: Connotations of drug delivery systems (DDSs) to target lymphatic filaroids or wolbachia and systemic microfilaria have been discussed. The potentials of liposomes and solid lipid nanoparticles for the treatment of LF are highlighted. Various critical factors, viz optimal size range, surface properties, preferred pH, mechanism of reticuloendothelial avoidance, and control of the release of antifilarial agents for safe elimination of parasites, are enclosed to design a novel DDS for LF. The review of nanotechnological approaches to improve antifilarial chemotherapy will help to resolve existing technological gaps.

EXPERT OPINION: Precincts in the antifilarial discovery programs can never be overcome by conventional methods. Nanomedicine encompasses wide-range solution for each single problem (i.e., from poor solubility to nonspecific targeting of antifilarial agents) for the cure of LF at low costs and may reduce the economic burden of LF diseases. Advances in nanotechnology loom will certainly come forward as silver bullets in the near future for quick diagnosis, control, and elimination of this tropically neglected disease.}, } @article {pmid23425244, year = {2013}, author = {Tabar, MD and Altet, L and Martínez, V and Roura, X}, title = {Wolbachia, filariae and Leishmania coinfection in dogs from a Mediterranean area.}, journal = {The Journal of small animal practice}, volume = {54}, number = {4}, pages = {174-178}, doi = {10.1111/jsap.12041}, pmid = {23425244}, issn = {1748-5827}, mesh = {Animals ; Coinfection/epidemiology/microbiology/parasitology/*veterinary ; Dog Diseases/*epidemiology/microbiology/parasitology ; Dogs ; Female ; Filariasis/epidemiology/*veterinary ; Leishmaniasis/epidemiology/*veterinary ; Male ; Mediterranean Region ; Polymerase Chain Reaction/veterinary ; Spain/epidemiology ; *Wolbachia ; }, abstract = {OBJECTIVES: In an endemic area for leishmaniosis and filariasis, coinfection can occur and the immunomodulation triggered by Wolbachia infection might influence the clinical signs and progression of both diseases. The aims of this study were to determine the prevalence of Wolbachia in dogs infected with Dirofilaria immitis and other filarial nematodes, to evaluate the prevalence of coinfection of Leishmania infantum, filariae and Wolbachia and their association with clinical presentation.

METHODS: Polymerase chain reaction assays were performed to detect filarial species, Wolbachia species and Leishmania in 118 samples of dogs from southeastern Spain with leishmaniosis and/or filariasis.

RESULTS: Ninety-eight dogs were infected with Leishmania and 49 had filarial infection (29 were coinfected with both). Wolbachia DNA was detected in 30·6% of filariae-positive dogs (15/49). Dogs coinfected with Leishmania and filaria had more severe clinical signs. Wolbachia infection was significantly (P=0·026) more frequent in dogs that were not infected with Leishmania. There was no correlation between outcome and coinfection with these pathogens.

CLINICAL SIGNIFICANCE: This study highlights the increased sensitivity of polymerase chain reaction in the diagnosis of filariasis, confirms the presence of Wolbachia in dogs from the Mediterranean basin, shows the increased severity of clinical signs when Leishmania-filarial coinfection is present and suggests a protective role of Wolbachia in leishmaniosis.}, } @article {pmid23413179, year = {2012}, author = {Almerão, MP and Fagundes, NJ and de Araújo, PB and Verne, S and Grandjean, F and Bouchon, D and Araújo, AM}, title = {First record of Wolbachia in South American terrestrial isopods: Prevalence and diversity in two species of Balloniscus (Crustacea, Oniscidea).}, journal = {Genetics and molecular biology}, volume = {35}, number = {4 (suppl)}, pages = {980-989}, pmid = {23413179}, issn = {1415-4757}, abstract = {Wolbachia are endosymbiotic bacteria that commonly infect arthropods, inducing certain phenotypes in their hosts. So far, no endemic South American species of terrestrial isopods have been investigated for Wolbachia infection. In this work, populations from two species of Balloniscus (B. sellowii and B. glaber) were studied through a diagnostic PCR assay. Fifteen new Wolbachia 16S rDNA sequences were detected. Wolbachia found in both species were generally specific to one population, and five populations hosted two different Wolbachia 16S rDNA sequences. Prevalence was higher in B. glaber than in B. sellowii, but uninfected populations could be found in both species. Wolbachia strains from B. sellowii had a higher genetic variation than those isolated from B. glaber. AMOVA analyses showed that most of the genetic variance was distributed among populations of each species rather than between species, and the phylogenetic analysis suggested that Wolbachia strains from Balloniscus cluster within Supergroup B, but do not form a single monophyletic clade, suggesting multiple infections for this group. Our results highlight the importance of studying Wolbachia prevalence and genetic diversity in Neotropical species and suggest that South American arthropods may harbor a great number of diverse strains, providing an interesting model to investigate the evolution of Wolbachia and its hosts.}, } @article {pmid23404314, year = {2012}, author = {Kvie, KS and Hogner, S and Aarvik, L and Lifjeld, JT and Johnsen, A}, title = {Deep sympatric mtDNA divergence in the autumnal moth (Epirrita autumnata).}, journal = {Ecology and evolution}, volume = {3}, number = {1}, pages = {126-144}, pmid = {23404314}, issn = {2045-7758}, abstract = {Deep sympatric intraspecific divergence in mtDNA may reflect cryptic species or formerly distinct lineages in the process of remerging. Preliminary results from DNA barcoding of Scandinavian butterflies and moths showed high intraspecific sequence variation in the autumnal moth, Epirrita autumnata. In this study, specimens from different localities in Norway and some samples from Finland and Scotland, with two congeneric species as outgroups, were sequenced with mitochondrial and nuclear markers to resolve the discrepancy found between mtDNA divergence and present species-level taxonomy. We found five COI sub-clades within the E. autumnata complex, most of which were sympatric and with little geographic structure. Nuclear markers (ITS2 and Wingless) showed little variation and gave no indications that E. autumnata comprises more than one species. The samples were screened with primers for Wolbachia outer surface gene (wsp) and 12% of the samples tested positive. Two Wolbachia strains were associated with different mtDNA sub-clades within E. autumnata, which may indicate indirect selection/selective sweeps on haplotypes. Our results demonstrate that deep mtDNA divergences are not synonymous with cryptic speciation and this has important implications for the use of mtDNA in species delimitation, like in DNA barcoding.}, } @article {pmid23399027, year = {2013}, author = {Turley, AP and Zalucki, MP and O'Neill, SL and McGraw, EA}, title = {Transinfected Wolbachia have minimal effects on male reproductive success in Aedes aegypti.}, journal = {Parasites & vectors}, volume = {6}, number = {}, pages = {36}, pmid = {23399027}, issn = {1756-3305}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Female ; Fertility ; Genetic Fitness ; Larva/physiology ; Male ; Survival Analysis ; Wolbachia/*growth & development ; }, abstract = {BACKGROUND: Wolbachia are maternally inherited endosymbiotic bacteria that manipulate the reproductive success of their insect hosts. Uninfected females that mate with Wolbachia infected males do not reproduce due to cytoplasmic incompatibility (CI). CI results in the increased frequency of Wolbachia-infected individuals in populations. Recently, two Wolbachia strains, the benign wMel and virulent wMelPop have been artificially transinfected into the primary vector of dengue virus, the mosquito Ae. aegypti where they have formed stable infections. These Wolbachia infections are being developed for a biological control strategy against dengue virus transmission. While the effects of Wolbachia on female Ae. aegypti have been examined the effects on males are less well characterised. Here we ascertain and compare the effects of the two strains on male fitness in resource-limited environments that may better approximate the natural environment.

METHODS: A series of population mating trials were conducted to examine the effect of Wolbachia infection status (with strains wMel and wMelPop) and male larval nutrition on insemination frequency, remating rates, the fecundity of females, the hatch rates of eggs and the wing length and fertility of males.

RESULTS: wMel and wMelPop infections reduce the fecundity of infected females and wMelPop reduces the viability of eggs. Low nutrition diets for males in the larval phase affects the fecundity of wMel-infected females. Neither strain of Wolbachia affected sperm quality or viability or the ability of males to successfully mate multiple females.

CONCLUSIONS: The benign strain of Wolbachia, wMel causes similar reductions in fecundity as the more virulent, wMelPop, and neither are too great that they should not still spread given the action of CI. The ability of Wolbachia-infected males to repeat mate as frequently as wildtype mosquitoes indicates that they will be very good agents of delivering CI in field release populations.}, } @article {pmid23398406, year = {2013}, author = {Bouchery, T and Lefoulon, E and Karadjian, G and Nieguitsila, A and Martin, C}, title = {The symbiotic role of Wolbachia in Onchocercidae and its impact on filariasis.}, journal = {Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases}, volume = {19}, number = {2}, pages = {131-140}, doi = {10.1111/1469-0691.12069}, pmid = {23398406}, issn = {1469-0691}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Filariasis/drug therapy ; Filaricides/therapeutic use ; Filarioidea/*microbiology ; Humans ; *Symbiosis ; Tetracycline/*therapeutic use ; Wolbachia/*drug effects/*physiology ; }, abstract = {Symbiotic associations between eukaryotes and microorganisms are frequently observed in nature, and range along the continuum between parasitism and mutualism. The genus Wolbachia contains well-known intracellular bacteria of arthropods that induce several reproductive phenotypes that benefit the transmission of the bacteria. Interestingly, Wolbachia bacteria have been found in the Onchocercidae, a family of filarial nematodes, including species that cause human filarial diseases, e.g. lymphatic filariasis and onchocerciasis. The endosymbiont is thought to be mutualistic in the Onchocercidae, and to provide essential metabolites to the filariae. Currently, Wolbachia bacteria are targets of antibiotic therapy with tetracyclines, which have profound effects on the development, viability and fertility of filarial parasites. This overview article presents the Onchocercidae and Wolbachia, and then discusses the origin and the nature of the symbiosis. It highlights the contribution of Wolbachia to the survival of the filariae and to the development of pathology. Finally, the infection control implications for filariases are debated. Potential directions for future research are also discussed.}, } @article {pmid23398404, year = {2013}, author = {Cutler, SJ}, title = {Dangerous liaisons--our expanding appreciation of microbial synergism.}, journal = {Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases}, volume = {19}, number = {2}, pages = {105-106}, doi = {10.1111/1469-0691.12066}, pmid = {23398404}, issn = {1469-0691}, mesh = {Anti-Bacterial Agents/*therapeutic use ; Coinfection/*epidemiology/microbiology ; Communicable Diseases/*epidemiology/etiology ; Humans ; Immunologic Factors/*therapeutic use ; Leishmaniasis/*complications/drug therapy ; RNA Viruses/immunology/physiology ; Superinfection/*epidemiology/etiology ; Symbiosis ; Wolbachia/drug effects/physiology ; }, } @article {pmid23390535, year = {2013}, author = {Siozios, S and Ioannidis, P and Klasson, L and Andersson, SG and Braig, HR and Bourtzis, K}, title = {The diversity and evolution of Wolbachia ankyrin repeat domain genes.}, journal = {PloS one}, volume = {8}, number = {2}, pages = {e55390}, pmid = {23390535}, issn = {1932-6203}, mesh = {Animals ; Ankyrin Repeat/*genetics ; Ankyrins/chemistry/*genetics ; Bacterial Proteins/chemistry/*genetics ; Bacteriophages/genetics ; Base Sequence ; Drosophila/classification/microbiology ; Evolution, Molecular ; Gene Transfer, Horizontal ; *Genetic Variation ; *Genome, Bacterial ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Symbiosis/*genetics ; Wolbachia/classification/*genetics/metabolism ; }, abstract = {Ankyrin repeat domain-encoding genes are common in the eukaryotic and viral domains of life, but they are rare in bacteria, the exception being a few obligate or facultative intracellular Proteobacteria species. Despite having a reduced genome, the arthropod strains of the alphaproteobacterium Wolbachia contain an unusually high number of ankyrin repeat domain-encoding genes ranging from 23 in wMel to 60 in wPip strain. This group of genes has attracted considerable attention for their astonishing large number as well as for the fact that ankyrin proteins are known to participate in protein-protein interactions, suggesting that they play a critical role in the molecular mechanism that determines host-Wolbachia symbiotic interactions. We present a comparative evolutionary analysis of the wMel-related ankyrin repeat domain-encoding genes present in different Drosophila-Wolbachia associations. Our results show that the ankyrin repeat domain-encoding genes change in size by expansion and contraction mediated by short directly repeated sequences. We provide examples of intra-genic recombination events and show that these genes are likely to be horizontally transferred between strains with the aid of bacteriophages. These results confirm previous findings that the Wolbachia genomes are evolutionary mosaics and illustrate the potential that these bacteria have to generate diversity in proteins potentially involved in the symbiotic interactions.}, } @article {pmid23384159, year = {2013}, author = {Symula, RE and Alam, U and Brelsfoard, C and Wu, Y and Echodu, R and Okedi, LM and Aksoy, S and Caccone, A}, title = {Wolbachia association with the tsetse fly, Glossina fuscipes fuscipes, reveals high levels of genetic diversity and complex evolutionary dynamics.}, journal = {BMC evolutionary biology}, volume = {13}, number = {}, pages = {31}, pmid = {23384159}, issn = {1471-2148}, support = {D43 TW007391/TW/FIC NIH HHS/United States ; R03 TW008755/TW/FIC NIH HHS/United States ; AI068932/AI/NIAID NIH HHS/United States ; D43TW007391/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; Chaperonin 60/genetics ; DNA, Mitochondrial/genetics ; Female ; Genes, Bacterial ; *Genetic Variation ; *Genetics, Population ; Geography ; Haplotypes ; Likelihood Functions ; Multilocus Sequence Typing ; Phylogeny ; Tsetse Flies/genetics/*microbiology ; Uganda ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Wolbachia pipientis, a diverse group of α-proteobacteria, can alter arthropod host reproduction and confer a reproductive advantage to Wolbachia-infected females (cytoplasmic incompatibility (CI)). This advantage can alter host population genetics because Wolbachia-infected females produce more offspring with their own mitochondrial DNA (mtDNA) haplotypes than uninfected females. Thus, these host haplotypes become common or fixed (selective sweep). Although simulations suggest that for a CI-mediated sweep to occur, there must be a transient phase with repeated initial infections of multiple individual hosts by different Wolbachia strains, this has not been observed empirically. Wolbachia has been found in the tsetse fly, Glossina fuscipes fuscipes, but it is not limited to a single host haplotype, suggesting that CI did not impact its population structure. However, host population genetic differentiation could have been generated if multiple Wolbachia strains interacted in some populations. Here, we investigated Wolbachia genetic variation in G. f. fuscipes populations of known host genetic composition in Uganda. We tested for the presence of multiple Wolbachia strains using Multi-Locus Sequence Typing (MLST) and for an association between geographic region and host mtDNA haplotype using Wolbachia DNA sequence from a variable locus, groEL (heat shock protein 60).

RESULTS: MLST demonstrated that some G. f. fuscipes carry Wolbachia strains from two lineages. GroEL revealed high levels of sequence diversity within and between individuals (Haplotype diversity = 0.945). We found Wolbachia associated with 26 host mtDNA haplotypes, an unprecedented result. We observed a geographical association of one Wolbachia lineage with southern host mtDNA haplotypes, but it was non-significant (p = 0.16). Though most Wolbachia-infected host haplotypes were those found in the contact region between host mtDNA groups, this association was non-significant (p = 0.17).

CONCLUSIONS: High Wolbachia sequence diversity and the association of Wolbachia with multiple host haplotypes suggest that different Wolbachia strains infected G. f. fuscipes multiple times independently. We suggest that these observations reflect a transient phase in Wolbachia evolution that is influenced by the long gestation and low reproductive output of tsetse. Although G. f. fuscipes is superinfected with Wolbachia, our data does not support that bidirectional CI has influenced host genetic diversity in Uganda.}, } @article {pmid23383354, year = {2013}, author = {Chen, L and Zhu, C and Zhang, D}, title = {Naturally occurring incompatibilities between different Culex pipiens pallens populations as the basis of potential mosquito control measures.}, journal = {PLoS neglected tropical diseases}, volume = {7}, number = {1}, pages = {e2030}, pmid = {23383354}, issn = {1935-2735}, mesh = {Animals ; China ; Crosses, Genetic ; Culex/genetics/*microbiology/physiology ; Female ; Germ Cells/*microbiology ; Male ; Molecular Sequence Data ; Mosquito Control/*methods ; Reproduction ; Sequence Analysis, DNA ; Sexual Behavior ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Vector-borne diseases remain a threat to public health, especially in tropical countries. The incompatible insect technique has been explored as a potential control strategy for several important insect vectors. However, this strategy has not been tested in Culex pipiens pallens, the most prevalent mosquito species in China. Previous works used introgression to generate new strains that matched the genetic backgrounds of target populations while harboring a new Wolbachia endosymbiont, resulting in mating competitiveness and cytoplasmic incompatibility. The generation of these incompatible insects is often time-consuming, and the long-term stability of the newly created insect-Wolbachia symbiosis is uncertain. Considering the wide distribution of Cx. pipiens pallens and hence possible isolation of different populations, we sought to test for incompatibilities between natural populations and the possibility of exploiting these incompatibilities as a control strategy.

Three field populations were collected from three geographic locations in eastern China. Reciprocal cross results showed that bi-directional patterns of incompatibility existed between some populations. Mating competition experiments indicated that incompatible males could compete with cognate males in mating with females, leading to reduced overall fecundity. F1 offspring from incompatible crosses maintained their maternal crossing types. All three populations tested positive for Wolbachia. Removal of Wolbachia by tetracycline rendered matings between these populations fully compatible.

CONCLUSIONS/SIGNIFICANCE: Our findings indicate that naturally occurring patterns of cytoplasmic incompatibility between Cx. pipiens pallens populations can be the basis of a control strategy for this important vector species. The observed incompatibilities are caused by Wolbachia. More tests including field trials are warranted to evaluate the feasibility of this strategy as a supplement to other control measures.}, } @article {pmid23379718, year = {2013}, author = {Merville, A and Venner, S and Henri, H and Vallier, A and Menu, F and Vavre, F and Heddi, A and Bel-Venner, MC}, title = {Endosymbiont diversity among sibling weevil species competing for the same resource.}, journal = {BMC evolutionary biology}, volume = {13}, number = {}, pages = {28}, pmid = {23379718}, issn = {1471-2148}, mesh = {Animals ; Ecosystem ; Female ; France ; Male ; Molecular Sequence Data ; Phylogeny ; *Quercus ; Rickettsia/classification/physiology ; Rickettsiaceae/classification/*physiology ; Spiroplasma/classification/*physiology ; *Symbiosis ; Weevils/classification/*microbiology/*physiology ; Wolbachia/classification/physiology ; }, abstract = {BACKGROUND: Whereas the impact of endosymbionts on the ecology of their hosts is well known in some insect species, the question of whether host communities are influenced by endosymbionts remains largely unanswered. Notably, the coexistence of host species competing with each other, which is expected to be stabilized by their ecological differences, could be facilitated by differences in their endosymbionts. Yet, the composition of endosymbiotic communities housed by natural communities of competing host species is still almost unknown. In this study, we started filling this gap by describing and comparing the bacterial endosymbiotic communities of four sibling weevil species (Curculio spp.) that compete with each other to lay eggs into oak acorns (Quercus spp.) and exhibit marked ecological differences.

RESULTS: All four species housed the primary endosymbiont Candidatus Curculioniphilus buchneri, yet each of these had a clearly distinct community of secondary endosymbionts, including Rickettsia, Spiroplasma, and two Wolbachia strains. Notably, three weevil species harbored their own predominant facultative endosymbiont and possessed the remaining symbionts at a residual infection level.

CONCLUSIONS: The four competing species clearly harbor distinct endosymbiotic communities. We discuss how such endosymbiotic communities could spread and keep distinct in the four insect species, and how these symbionts might affect the organization and species richness of host communities.}, } @article {pmid23355904, year = {2013}, author = {Zhao, DX and Chen, DS and Ge, C and Gotoh, T and Hong, XY}, title = {Multiple infections with Cardinium and two strains of Wolbachia in the spider mite Tetranychus phaselus Ehara: revealing new forces driving the spread of Wolbachia.}, journal = {PloS one}, volume = {8}, number = {1}, pages = {e54964}, pmid = {23355904}, issn = {1932-6203}, mesh = {Animals ; *Bacterial Infections/metabolism/microbiology/transmission ; Bacteroidaceae/*metabolism ; Coinfection/*metabolism/microbiology ; Female ; Male ; Tetranychidae/*metabolism/*microbiology ; Wolbachia/*metabolism ; }, abstract = {Cytoplasmic incompatibility (CI) has been proposed as a major mechanism by which certain strains of Wolbachia to invade and persist in host populations. However, mechanisms that underlie the invasion and persistence of non-CI strains are less well understood. Here, we established a spider mite Tetranychus phaselus population multiply infected by Cardinium as well as two distinct lineages of Wolbachia, designated wCon and wOri, to study the forces driving the spread of the non-CI strain of Wolbachia wOri. Interestingly, we found that wOri provided a longevity advantage to its female hosts under ideal conditions, making wOri stay longer in this population, and then being transmitted to more offspring. Furthermore, the lifespan of uninfected females was reduced when mated with multiple-infected males. As a result, the uninfected population is attenuated by the multiple-infected males. Thus, we infer that the host age effects of multiple infection may represent sufficient forces driving the spread of wOri through the host population.}, } @article {pmid23349865, year = {2013}, author = {Ilinsky, Y}, title = {Coevolution of Drosophila melanogaster mtDNA and Wolbachia genotypes.}, journal = {PloS one}, volume = {8}, number = {1}, pages = {e54373}, pmid = {23349865}, issn = {1932-6203}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Drosophila melanogaster/*genetics ; Evolution, Molecular ; Female ; Genetic Variation ; Genotype ; Haplotypes ; Host-Parasite Interactions/*genetics ; Mitochondria/genetics ; Phylogeny ; Wolbachia/*genetics ; }, abstract = {Maternally inherited microorganisms can influence the mtDNA pattern of variation in hosts. This influence is driven by selection among symbionts and can cause the frequency of mitochondrial variants in the population to eventually increase or decrease. Wolbachia infection is common and widespread in Drosophila melanogaster populations. We compared genetic variability of D. melanogaster mitotypes with Wolbachia genotypes among isofemale lines associated with different geographic locations and time intervals to study coevolution of the mtDNA and Wolbachia. Phylogenetic analysis of D. melanogaster mtDNA revealed two clades diverged in Africa, each associated with one of the two Wolbachia genotype groups. No evidence of horizontal transmission of Wolbachia between maternal lineages has been found. All the mtDNA variants that occur in infected isofemale lines are found in uninfected isofemale lines and vice versa, which is indicative of a recent loss of infection from some maternal fly lineages and confirms a significant role of Wolbachia in the D. melanogaster mtDNA pattern of variation. Finally, we present a comparative analysis of biogeographic distribution of D. melanogaster mitotypes all over the world.}, } @article {pmid23349831, year = {2013}, author = {Elnagdy, S and Messing, S and Majerus, ME}, title = {Two strains of male-killing Wolbachia in a ladybird, Coccinella undecimpunctata, from a hot climate.}, journal = {PloS one}, volume = {8}, number = {1}, pages = {e54218}, pmid = {23349831}, issn = {1932-6203}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Bacterial Outer Membrane Proteins/genetics ; *Climate ; Coleoptera/drug effects/*microbiology ; Egypt ; Female ; Host-Pathogen Interactions/drug effects ; *Hot Temperature ; Jordan ; Male ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Sex Factors ; Sex Ratio ; Species Specificity ; Tetracycline/pharmacology ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Ladybirds are a hot-spot for the invasion of male-killing bacteria. These maternally inherited endosymbionts cause the death of male host embryos, to the benefit of female sibling hosts and the bacteria that they contain. Previous studies have shown that high temperatures can eradicate male-killers from ladybirds, leaving the host free from infection. Here we report the discovery of two maternally inherited sex ratio distorters in populations of a coccinellid, Coccinella undecimpunctata, from a hot lowland region of the Middle East. DNA sequence analysis indicates that the male killing is the result of infection by Wolbachia, that the trait is tetracycline sensitive, and that two distinct strains of Wolbachia co-occur within one beetle population. We discuss the implications of these findings for theories of male-killing and suggest avenues for future field-work on this system.}, } @article {pmid23334948, year = {2013}, author = {Ahantarig, A and Trinachartvanit, W and Baimai, V and Grubhoffer, L}, title = {Hard ticks and their bacterial endosymbionts (or would be pathogens).}, journal = {Folia microbiologica}, volume = {58}, number = {5}, pages = {419-428}, pmid = {23334948}, issn = {1874-9356}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; *Bacterial Physiological Phenomena ; *Biodiversity ; Ixodidae/*microbiology/physiology ; *Symbiosis ; }, abstract = {The symbiotic microorganisms of arthropod vectors are highly significant from several points of view, partly due to their possible roles in the transmission of pathogenic causative agents by blood-sucking vectors. Although ticks are well studied because of their significance to human health, novel microbial associations remain to be described. This review summarises several endosymbiotic bacterial species in hard ticks from various parts of the world, including Coxiella-, Francisella-, Rickettsia- and Arsenophonus-like symbionts as well as Candidatus Midichloria mitochondrii and Wolbachia. New methodologies for the isolation and characterization of tick-associated bacteria will, in turn, encourage new strategies of tick control by studying their endosymbionts.}, } @article {pmid23331872, year = {2013}, author = {Patricelli, D and Sielezniew, M and Ponikwicka-Tyszko, D and Ratkiewicz, M and Bonelli, S and Barbero, F and Witek, M and Buś, MM and Rutkowski, R and Balletto, E}, title = {Contrasting genetic structure of rear edge and continuous range populations of a parasitic butterfly infected by Wolbachia.}, journal = {BMC evolutionary biology}, volume = {13}, number = {}, pages = {14}, pmid = {23331872}, issn = {1471-2148}, mesh = {Animals ; Butterflies/*genetics/microbiology ; Cell Nucleus/genetics ; DNA, Mitochondrial/genetics ; *Genetic Variation ; *Genetics, Population ; Geography ; Haplotypes ; Italy ; Poland ; Polymorphism, Genetic ; Sequence Analysis, DNA ; *Wolbachia ; }, abstract = {BACKGROUND: Climatic oscillations are among the long-term factors shaping the molecular features of animals and plants and it is generally supposed that the rear edges (i.e., the low-latitude limits of distribution of any given specialised species) situated closer to glacial refugia are vital long-term stores of genetic diversity. In the present study, we compared the genetic structure of several populations of an endangered and obligate myrmecophilous butterfly (Maculinea arion) from two distinct and geographically distant parts of its European distribution (i.e., Italy and Poland), which fully represent the ecological and morphological variation occurring across the continent.

RESULTS: We sequenced the COI mitochondrial DNA gene (the 'barcoding gene') and the EF-1α nuclear gene and found substantial genetic differentiation among M. arion Italian populations in both markers. Eleven mtDNA haplotypes were present in Italy. In contrast, almost no mtDNA polymorphisms was found in the Polish M. arion populations, where genetic differentiation at the nuclear gene was low to moderate. Interestingly, the within-population diversity levels in the EF-1α gene observed in Italy and in Poland were comparable. The genetic data did not support any subspecies divisions or any ecological specialisations. All of the populations studied were infected with a single strain of Wolbachia and our screening suggested 100% prevalence of the bacterium.

CONCLUSIONS: Differences in the genetic structure of M. arion observed in Italy and in Poland may be explained by the rear edge theory. Although we were not able to pinpoint any specific evolutionarily significant units, we suggest that the Italian peninsula should be considered as a region of special conservation concern and one that is important for maintaining the genetic diversity of M. arion in Europe. The observed pattern of mtDNA differentiation among the populations could not be explained by an endosymbiotic infection.}, } @article {pmid23324387, year = {2013}, author = {Duplouy, A and Iturbe-Ormaetxe, I and Beatson, SA and Szubert, JM and Brownlie, JC and McMeniman, CJ and McGraw, EA and Hurst, GD and Charlat, S and O'Neill, SL and Woolfit, M}, title = {Draft genome sequence of the male-killing Wolbachia strain wBol1 reveals recent horizontal gene transfers from diverse sources.}, journal = {BMC genomics}, volume = {14}, number = {}, pages = {20}, pmid = {23324387}, issn = {1471-2164}, mesh = {Adenosine Triphosphatases/genetics ; Animals ; Bacterial Proteins/genetics ; Butterflies/microbiology ; *Gene Transfer, Horizontal ; Genome, Bacterial/*genetics ; Male ; Membrane Transport Proteins/genetics ; Molecular Sequence Annotation ; Phylogeny ; SEC Translocation Channels ; SecA Proteins ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: The endosymbiont Wolbachia pipientis causes diverse and sometimes dramatic phenotypes in its invertebrate hosts. Four Wolbachia strains sequenced to date indicate that the constitution of the genome is dynamic, but these strains are quite divergent and do not allow resolution of genome diversification over shorter time periods. We have sequenced the genome of the strain wBol1-b, found in the butterfly Hypolimnas bolina, which kills the male offspring of infected hosts during embyronic development and is closely related to the non-male-killing strain wPip from Culex pipiens.

RESULTS: The genomes of wBol1-b and wPip are similar in genomic organisation, sequence and gene content, but show substantial differences at some rapidly evolving regions of the genome, primarily associated with prophage and repetitive elements. We identified 44 genes in wBol1-b that do not have homologs in any previously sequenced strains, indicating that Wolbachia's non-core genome diversifies rapidly. These wBol1-b specific genes include a number that have been recently horizontally transferred from phylogenetically distant bacterial taxa. We further report a second possible case of horizontal gene transfer from a eukaryote into Wolbachia.

CONCLUSIONS: Our analyses support the developing view that many endosymbiotic genomes are highly dynamic, and are exposed and receptive to exogenous genetic material from a wide range of sources. These data also suggest either that this bacterial species is particularly permissive for eukaryote-to-prokaryote gene transfers, or that these transfers may be more common than previously believed. The wBol1-b-specific genes we have identified provide candidates for further investigations of the genomic bases of phenotypic differences between closely-related Wolbachia strains.}, } @article {pmid23308142, year = {2013}, author = {Ahmed, MZ and De Barro, PJ and Ren, SX and Greeff, JM and Qiu, BL}, title = {Evidence for horizontal transmission of secondary endosymbionts in the Bemisia tabaci cryptic species complex.}, journal = {PloS one}, volume = {8}, number = {1}, pages = {e53084}, pmid = {23308142}, issn = {1932-6203}, mesh = {Animals ; Bacteroidetes/*genetics/physiology ; Biological Evolution ; Enterobacteriaceae/*genetics/physiology ; Hemiptera/*genetics/*microbiology/physiology ; Phylogeny ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {Bemisia tabaci (Hemiptera: Aleyrodidae) is a globally distributed pest composed of at least 34 morphologically indistinguishable cryptic species. At least seven species of endosymbiont have been found infecting some or all members of the complex. The origin(s) of the associations between specific endosymbionts and their whitefly hosts is unknown. Infection is normally vertical, but horizontal transmission does occur and is one way for new infections to be introduced into individuals. The relationships between the different members of the cryptic species complex and the endosymbionts have not been well explored. In this study, the phylogenies of different cryptic species of the host with those of their endosymbionts were compared. Of particular interest was whether there was evidence for both coevolution and horizontal transmission. Congruence was observed for the primary endosymbiont, Portiera aleyrodidarum, and partial incongruence in the case of two secondary endosymbionts, Arsenophonus and Cardinium and incongruence for a third, Wolbachia. The patterns observed for the primary endosymbiont supported cospeciation with the host while the patterns for the secondary endosymbionts, and especially Wolbachia showed evidence of host shifts and extinctions through horizontal transmission rather than cospeciation. Of particular note is the observation of several very recent host shift events in China between exotic invader and indigenous members of the complex. These shifts were from indigenous members of the complex to the invader as well as from the invader to indigenous relatives.}, } @article {pmid23301109, year = {2012}, author = {Mousson, L and Zouache, K and Arias-Goeta, C and Raquin, V and Mavingui, P and Failloux, AB}, title = {The native Wolbachia symbionts limit transmission of dengue virus in Aedes albopictus.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {12}, pages = {e1989}, pmid = {23301109}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*virology ; Animal Structures/microbiology/virology ; Animals ; Bacterial Load ; Dengue Virus/*growth & development/physiology ; *Disease Vectors ; Female ; Humans ; Mice ; *Microbial Interactions ; Real-Time Polymerase Chain Reaction ; Reunion ; Saliva/virology ; *Symbiosis ; Viral Load ; Virus Replication ; Wolbachia/growth & development/*physiology ; }, abstract = {BACKGROUND: The chikungunya (CHIK) outbreak that struck La Reunion Island in 2005 was preceded by few human cases of Dengue (DEN), but which surprisingly did not lead to an epidemic as might have been expected in a non-immune population. Both arboviral diseases are transmitted to humans by two main mosquito species, Aedes aegypti and Aedes albopictus. In the absence of the former, Ae. albopictus was the only species responsible for viral transmission on La Reunion Island. This mosquito is naturally super-infected with two Wolbachia strains, wAlbA and wAlbB. While Wolbachia does not affect replication of CHIK virus (CHIKV) in Ae. albopictus, a similar effect was not observed with DEN virus (DENV).

METHODS/PRINCIPAL FINDINGS: To understand the weak vectorial status of Ae. albopictus towards DENV, we used experimental oral infections of mosquitoes from La Reunion Island to characterize the impact of Wolbachia on DENV infection. Viral loads and Wolbachia densities were measured by quantitative PCR in different organs of Ae. albopictus where DENV replication takes place after ingestion. We found that: (i) Wolbachia does not affect viral replication, (ii) Wolbachia restricts viral density in salivary glands, and (iii) Wolbachia limits transmission of DENV, as infectious viral particles were only detected in the saliva of Wolbachia-uninfected Ae. albopictus, 14 days after the infectious blood-meal.

CONCLUSIONS: We show that Wolbachia does not affect the replication of DENV in Ae. albopictus. However, Wolbachia is able to reduce viral infection of salivary glands and limit transmission, suggesting a role of Wolbachia in naturally restricting the transmission of DENV in Ae. albopictus from La Reunion Island. The extension of this conclusion to other Ae. albopictus populations should be investigated.}, } @article {pmid23287400, year = {2013}, author = {Beckmann, JF and Markowski, TW and Witthuhn, BA and Fallon, AM}, title = {Detection of the Wolbachia-encoded DNA binding protein, HU beta, in mosquito gonads.}, journal = {Insect biochemistry and molecular biology}, volume = {43}, number = {3}, pages = {272-279}, pmid = {23287400}, issn = {1879-0240}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/chemistry/*microbiology ; Amino Acid Sequence ; Animals ; Bacterial Proteins/*isolation & purification/metabolism ; Culex/chemistry/*microbiology ; DNA-Binding Proteins/*isolation & purification/metabolism ; Electrophoresis, Polyacrylamide Gel ; Female ; Gonads/chemistry ; Male ; Molecular Sequence Data ; Multigene Family ; Tandem Mass Spectrometry ; Wolbachia/*chemistry/genetics/metabolism ; }, abstract = {Wolbachia are obligate intracellular bacteria that cause cytoplasmic incompatibility in mosquitoes. In an incompatible cross, eggs of uninfected females fail to hatch when fertilized by sperm from infected males. We used polyacrylamide gel electrophoresis and tandem mass spectrometry to identify Wolbachia proteins in infected mosquito gonads. These included surface proteins with masses of 25 and 18 kDa and the DNA binding protein, HU beta. Using reverse transcriptase polymerase chain reaction, we showed that the HU gene is transcribed in Wolbachia-infected Culex pipiens and Aedes albopictus mosquitoes. We sequenced HU genes from four Wolbachia strains and compared deduced protein sequences with additional homologs from the databases. Among the Rickettsiales, Wolbachia HU has distinct N- and C-terminal basic/acidic amino acid motifs as well as a pair of conserved, cysteine residues.}, } @article {pmid23284655, year = {2012}, author = {Russell, JA and Funaro, CF and Giraldo, YM and Goldman-Huertas, B and Suh, D and Kronauer, DJ and Moreau, CS and Pierce, NE}, title = {A veritable menagerie of heritable bacteria from ants, butterflies, and beyond: broad molecular surveys and a systematic review.}, journal = {PloS one}, volume = {7}, number = {12}, pages = {e51027}, pmid = {23284655}, issn = {1932-6203}, mesh = {Animals ; Ants/*microbiology ; Bacteria/classification/*genetics ; *Bacterial Physiological Phenomena ; Butterflies/*microbiology ; Host Specificity ; Phylogeny ; }, abstract = {Maternally transmitted bacteria have been important players in the evolution of insects and other arthropods, affecting their nutrition, defense, development, and reproduction. Wolbachia are the best studied among these and typically the most prevalent. While several other bacteria have independently evolved a heritable lifestyle, less is known about their host ranges. Moreover, most groups of insects have not had their heritable microflora systematically surveyed across a broad range of their taxonomic diversity. To help remedy these shortcomings we used diagnostic PCR to screen for five groups of heritable symbionts-Arsenophonus spp., Cardinium hertigii, Hamiltonella defensa, Spiroplasma spp., and Wolbachia spp.-across the ants and lepidopterans (focusing, in the latter case, on two butterfly families-the Lycaenidae and Nymphalidae). We did not detect Cardinium or Hamiltonella in any host. Wolbachia were the most widespread, while Spiroplasma (ants and lepidopterans) and Arsenophonus (ants only) were present at low levels. Co-infections with different Wolbachia strains appeared especially common in ants and less so in lepidopterans. While no additional facultative heritable symbionts were found among ants using universal bacterial primers, microbes related to heritable enteric bacteria were detected in several hosts. In summary, our findings show that Wolbachia are the dominant heritable symbionts of ants and at least some lepidopterans. However, a systematic review of symbiont frequencies across host taxa revealed that this is not always the case across other arthropods. Furthermore, comparisons of symbiont frequencies revealed that the prevalence of Wolbachia and other heritable symbionts varies substantially across lower-level arthropod taxa. We discuss the correlates, potential causes, and implications of these patterns, providing hypotheses on host attributes that may shape the distributions of these influential bacteria.}, } @article {pmid23284297, year = {2012}, author = {Richardson, MF and Weinert, LA and Welch, JJ and Linheiro, RS and Magwire, MM and Jiggins, FM and Bergman, CM}, title = {Population genomics of the Wolbachia endosymbiont in Drosophila melanogaster.}, journal = {PLoS genetics}, volume = {8}, number = {12}, pages = {e1003129}, pmid = {23284297}, issn = {1553-7404}, support = {/WT_/Wellcome Trust/United Kingdom ; WT094664/WT_/Wellcome Trust/United Kingdom ; BB/E012868/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Bayes Theorem ; *Drosophila melanogaster/genetics/physiology ; Evolution, Molecular ; Genetic Variation ; Genome, Mitochondrial ; Haplotypes ; *Metagenomics ; Phylogeny ; *Symbiosis ; *Wolbachia/genetics/physiology ; }, abstract = {Wolbachia are maternally inherited symbiotic bacteria, commonly found in arthropods, which are able to manipulate the reproduction of their host in order to maximise their transmission. The evolutionary history of endosymbionts like Wolbachia can be revealed by integrating information on infection status in natural populations with patterns of sequence variation in Wolbachia and host mitochondrial genomes. Here we use whole-genome resequencing data from 290 lines of Drosophila melanogaster from North America, Europe, and Africa to predict Wolbachia infection status, estimate relative cytoplasmic genome copy number, and reconstruct Wolbachia and mitochondrial genome sequences. Overall, 63% of Drosophila strains were predicted to be infected with Wolbachia by our in silico analysis pipeline, which shows 99% concordance with infection status determined by diagnostic PCR. Complete Wolbachia and mitochondrial genomes show congruent phylogenies, consistent with strict vertical transmission through the maternal cytoplasm and imperfect transmission of Wolbachia. Bayesian phylogenetic analysis reveals that the most recent common ancestor of all Wolbachia and mitochondrial genomes in D. melanogaster dates to around 8,000 years ago. We find evidence for a recent global replacement of ancestral Wolbachia and mtDNA lineages, but our data suggest that the derived wMel lineage arose several thousand years ago, not in the 20th century as previously proposed. Our data also provide evidence that this global replacement event is incomplete and is likely to be one of several similar incomplete replacement events that have occurred since the out-of-Africa migration that allowed D. melanogaster to colonize worldwide habitats. This study provides a complete genomic analysis of the evolutionary mode and temporal dynamics of the D. melanogaster-Wolbachia symbiosis, as well as important resources for further analyses of the impact of Wolbachia on host biology.}, } @article {pmid23281896, year = {2013}, author = {McNulty, SN and Fischer, K and Curtis, KC and Weil, GJ and Brattig, NW and Fischer, PU}, title = {Localization of Wolbachia-like gene transcripts and peptides in adult Onchocerca flexuosa worms indicates tissue specific expression.}, journal = {Parasites & vectors}, volume = {6}, number = {}, pages = {2}, pmid = {23281896}, issn = {1756-3305}, mesh = {Animals ; Bacterial Proteins/*biosynthesis ; Gene Expression Profiling ; Gene Transfer, Horizontal ; *Genes, Bacterial ; Immunohistochemistry ; In Situ Hybridization ; Onchocerca/*microbiology ; *Transcription, Genetic ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Most filarial species in the genus Onchocerca depend on Wolbachia endobacteria to successfully carry out their life cycle. O. flexuosa is a Wolbachia-free species, but its genome contains Wolbachia-like sequences presumably obtained from Wolbachia via horizontal gene transfer. Proteogenomic studies have shown that many of these Wolbachia-like sequences are expressed in adult worms.

METHODS: Six Wolbachia-like sequences in O. flexuosa were chosen for further study based on their sequence conservation with Wolbachia genes, length of predicted open reading frames, and expression at the RNA and/or protein levels. In situ hybridization and immunohistochemical labeling were used to localize Wolbachia-like transcripts and peptides in adult worm tissues.

RESULTS: RNA probes representing three of the six target sequences produced hybridization signals in worm tissues. These probes bound to transcripts in the intestine and lateral chords of both sexes, in the hypodermis, median chords and uteri in females, and in sperm precursor cells in males. Antibodies raised to three peptides corresponding to these transcripts bound to specific bands in a soluble extract of adult O. flexuosa by Western blot that were not labeled by control antibodies in pre-immune serum. Two of the three antibodies produced labeling patterns in adult worm sections that were similar to those of the RNA probes, while the third produced a different pattern.

CONCLUSIONS: A subset of the Wolbachia-like sequences present in the genome of the Wolbachia-free filarial species O. flexuosa are transcribed in tissues where Wolbachia reside in infected filarial species. Some of the peptides and/or proteins derived from these transcripts appear to be concentrated in the same tissues while others may be exported to other regions of the worm. These results suggest that horizontally transferred Wolbachia genes and gene products may replicate important Wolbachia functions in uninfected filarial worms.}, } @article {pmid23271364, year = {2013}, author = {Fallon, AM and Baldridge, GD and Higgins, LA and Witthuhn, BA}, title = {Wolbachia from the planthopper Laodelphax striatellus establishes a robust, persistent, streptomycin-resistant infection in clonal mosquito cells.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {49}, number = {1}, pages = {66-73}, pmid = {23271364}, issn = {1543-706X}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*cytology/microbiology ; Amino Acid Substitution/genetics ; Animals ; DNA Primers/genetics ; Drug Resistance, Bacterial/*genetics ; Hemiptera/*microbiology ; Mass Spectrometry ; Mutation, Missense/genetics ; Polymerase Chain Reaction ; Ribosomal Proteins/*genetics ; *Streptomycin ; Wolbachia/genetics/*physiology ; }, abstract = {The obligate intracellular bacterium, Wolbachia pipientis (Rickettsiales: Anaplasmataceae), distorts reproduction of its arthropod hosts to facilitate invasion of naïve populations. This property makes Wolbachia an attractive "gene drive" agent with potential applications in the control of insect vector populations. Genetic manipulation of Wolbachia will require in vitro systems for its propagation, genetic modification, amplification, and introduction into target insects. Here we show that Wolbachia from the planthopper, Laodelphax striatellus, establishes a robust infection in clonal C7-10 Aedes albopictus mosquito cells. Infected cells, designated C/wStr, expressed radiolabeled proteins that were enriched in cells grown in the absence of antibiotics that inhibit Wolbachia, relative to cultures grown in medium containing tetracycline and rifampicin. Using mass spectrometry, we verified that tryptic peptides from an upregulated 24 kDa band predominantly represented proteins encoded by the Wolbachia genome, including the outer surface protein, Wsp. We further showed that resistance of Wolbachia to streptomycin is associated with a K42R mutation in Wolbachia ribosomal protein S12, and that the pattern of amino acid substitutions in ribosomal protein S12 shows distinct differences in the closely related genera, Wolbachia and Rickettsia.}, } @article {pmid23269454, year = {2013}, author = {Guidolin, AS and Cônsoli, FL}, title = {Molecular characterization of Wolbachia strains associated with the invasive Asian citrus psyllid Diaphorina citri in Brazil.}, journal = {Microbial ecology}, volume = {65}, number = {2}, pages = {475-486}, pmid = {23269454}, issn = {1432-184X}, mesh = {Animals ; Bacterial Typing Techniques ; Brazil ; Citrus ; DNA, Bacterial/genetics ; DNA, Mitochondrial/genetics ; Genotype ; Haplotypes ; Hemiptera/genetics/*microbiology ; Host Specificity ; Insect Vectors/genetics/*microbiology ; Introduced Species ; Multilocus Sequence Typing ; Phylogeny ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*classification/genetics ; }, abstract = {Wolbachia is a symbiont intensively studied due to its ability to interfere with their host's reproduction, and it has been recently proposed as an alternative tool to control insect pests or vectors of diseases. The Asian citrus psyllid Diaphorina citri is an important pest of citrus since it vectors the bacterium that causes the "Huanglongbing" disease in citrus. The frequency and diversity of Wolbachia associated with D. citri is unknown, limiting the utilization of Wolbachia as an alternative strategy for insect management. Thus, we aimed to determine the natural rate of infection, to characterize the Wolbachia strains associated with this psyllid by "multilocus sequencing typing" (MLST) and wsp analysis, and to verify the association of the symbiont to particular genotypes of the host. Analysis indicated Wolbachia infects 100 % of all specimens tested from all 15 sampled populations. MLST revealed the occurrence of five new sequence types (STs) of Wolbachia, while analysis based on the wsp sequences indicated only four different types of Wolbachia. ST-173 was predominant, while the remaining STs were population specific. Analysis of the host-symbiont relationship did not reveal any particular association of Wolbachia and haplotypes or a decrease in nucleotide diversity of D. citri in populations in which more than one ST was recorded. The consequences of the diversity of STs reported are still unknown, but the fact that Wolbachia infection is fixed and that there is one ST with a broad distribution highlights the use of this symbiont as an alternative strategy to control D. citri.}, } @article {pmid23262214, year = {2013}, author = {Sharma, R and Hoti, SL and Vasuki, V and Sankari, T and Meena, RL and Das, PK}, title = {Filamentation temperature-sensitive protein Z (FtsZ) of Wolbachia, endosymbiont of Wuchereria bancrofti: a potential target for anti-filarial chemotherapy.}, journal = {Acta tropica}, volume = {125}, number = {3}, pages = {330-338}, doi = {10.1016/j.actatropica.2012.12.004}, pmid = {23262214}, issn = {1873-6254}, mesh = {Albendazole/administration & dosage ; Amino Acid Motifs ; Amino Acid Sequence ; Animals ; Anti-Infective Agents/administration & dosage ; Bacterial Proteins/antagonists & inhibitors/*genetics/*metabolism ; Cloning, Molecular ; Culex/microbiology ; Cytoskeletal Proteins/antagonists & inhibitors/*genetics/*metabolism ; Humans ; India ; Larva/microbiology ; Molecular Sequence Data ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Wolbachia/drug effects/*genetics/*metabolism ; Wuchereria bancrofti/isolation & purification/*microbiology ; }, abstract = {Lymphatic filariasis (LF) is a leading cause of morbidity in the tropical world. It is caused by the filarial parasites Wuchereria bancrofti, Brugia malayi and Brugia timori and transmitted by vector mosquitoes. Currently a programme for the elimination of LF, Global programme for Elimination of Lymphatic Filariasis (GPELF), is underway with the strategy of mass administration of single dose of diethylcarbamazine or ivermectin, in combination with an anthelmintic drug, albendazole. However, antifilarial drugs used in the programme are only microfilaricidal but not or only partially macrofilaricidal. Hence, there is a need to identify new targets for developing antifilarial drugs. Filarial parasites harbor rickettsial endosymbionts, Wolbachia sp., which play an important role in their biology and hence are considered as potential targets for antifilarial chemotherapy development. In this study, one of the cell division proteins of Wolbachia of the major lymphatic filarial parasite, W. bancrofti, viz., filamentation temperature-sensitive protein Z (FtsZ), was explored as a drug target. The gene coding for FtsZ protein was amplified from the genomic DNA of W. bancrofti, cloned and sequenced. The derived amino acid sequence of the gene revealed that FtsZ protein is 396 amino acids long and contained the tubulin motif (GGGTGTG) involved in GTP binding and the GTP hydrolyzing motif (NLDFAD). The FtsZ gene of endosymbiont showed limited sequence homology, but exhibited functional homology with β-tubulin of its host, W. bancrofti, as it had both the functional motifs and conserved amino acids that are critical for enzymatic activity. β-tubulin is the target for the anti-helminthic activity of albendazole and since FtsZ shares functional homology with, β-tubulin it may also be sensitive to albendazole. Therefore, the effect of albendazole was tested against Wolbachia occurring in mosquitoes instead of filarial parasites as the drug has lethal effect on the latter. Third instar larvae of Culex quinquefasciatus were treated with 0.25mg/ml of albendazole (test) or tetracycline (positive control) in the rearing medium for different intervals and tested for the presence of Wolbachia by FtsZ PCR. All the treated larvae were negative for the presence of the FtsZ band, whereas all the control larvae were positive. The findings of the study, thus indicated that FtsZ is sensitive to albendazole. In view of this albendazole appears to have dual targets; FtsZ in Wolbachia and β-tubulin in W. bancrofti. Further, the functional domain of the gene was assessed for polymorphism among recombinant clones representing 120 W. bancrofti parasites, prevalent across wide geographic areas of India and found to be highly conserved among them. Since it is highly conserved and plays an important role in Wolbachia cell division it appears to be a potential target for anti-filarial chemotherapy development.}, } @article {pmid23251587, year = {2012}, author = {Li, Z and Carlow, CK}, title = {Characterization of transcription factors that regulate the type IV secretion system and riboflavin biosynthesis in Wolbachia of Brugia malayi.}, journal = {PloS one}, volume = {7}, number = {12}, pages = {e51597}, pmid = {23251587}, issn = {1932-6203}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/chemistry/*metabolism ; *Bacterial Secretion Systems/drug effects ; Biosynthetic Pathways/drug effects ; Brugia malayi/drug effects/growth & development/*microbiology ; DNA, Intergenic/genetics ; Doxycycline/pharmacology ; Gene Expression Regulation, Bacterial/drug effects ; Genes, Reporter ; Host-Parasite Interactions/drug effects ; Humans ; Life Cycle Stages/drug effects ; Molecular Sequence Data ; Operon/genetics ; Promoter Regions, Genetic/genetics ; Protein Binding/drug effects/genetics ; Recombinant Proteins/metabolism ; Riboflavin/*biosynthesis ; Sequence Homology, Amino Acid ; Transcription Factors/chemistry/*metabolism ; Wolbachia/drug effects/genetics/*metabolism ; }, abstract = {The human filarial parasite Brugia malayi harbors an endosymbiotic bacterium Wolbachia (wBm) that is required for parasite survival. Consequently, targeting wBm is a promising approach for anti-filarial drug development. The Type IV secretion system (T4SS) plays an important role in bacteria-host interactions and is under stringent regulation by transcription factors. In wBm, most T4SS genes are contained in two operons. We show the wBm is active since the essential assembly factor virB8-1, is transcribed in adult worms and larval stages, and VirB8-1 is present in parasite lysates. We also identify two transcription factors (wBmxR1 and wBmxR2) that bind to the promoter region of several genes of the T4SS. Gel shift assays show binding of wBmxR1 to regions upstream of the virB9-2 and wBmxR2 genes, whereas wBmxR2 binds to virB4-2 and wBmxR1 promoter regions. Interestingly, both transcription factors bind to the promoter of the ribA gene that precedes virB8-1, the first gene in operon 1 of the wBm T4SS. RT-PCR reveals ribA and virB8-1 genes are co-transcribed as one operon, indicating the ribA gene and T4SS operon 1 are co-regulated by both wBmxR1 and wBmxR2. RibA encodes a bi-functional enzyme that catalyzes two essential steps in riboflavin (Vitamin B2) biosynthesis. Importantly, the riboflavin pathway is absent in B. malayi. We demonstrate the pathway is functional in wBm, and observe vitamin B2 supplementation partially rescues filarial parasites treated with doxycycline, indicating Wolbachia may supply the essential vitamin to its worm host. This is the first characterization of a transcription factor(s) from wBm and first report of co-regulation of genes of the T4SS and riboflavin biosynthesis pathway. In addition, our results demonstrate a requirement of vitamin B2 for worm health and fertility, and imply a nutritional role of the symbiont for the filarial parasite host.}, } @article {pmid23236534, year = {2012}, author = {Hamlin, KL and Moss, DM and Priest, JW and Roberts, J and Kubofcik, J and Gass, K and Streit, TG and Nutman, TB and Eberhard, ML and Lammie, PJ}, title = {Longitudinal monitoring of the development of antifilarial antibodies and acquisition of Wuchereria bancrofti in a highly endemic area of Haiti.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {12}, pages = {e1941}, pmid = {23236534}, issn = {1935-2735}, mesh = {Animals ; Antibodies, Helminth/*blood ; Antigens, Helminth/*blood ; Child ; Child, Preschool ; Elephantiasis, Filarial/*diagnosis/immunology/parasitology ; *Endemic Diseases ; Enzyme-Linked Immunosorbent Assay ; Epidemiologic Methods ; Female ; Haiti/epidemiology ; Humans ; Infant ; Longitudinal Studies ; Male ; Microscopy ; *Parasitemia ; Time Factors ; Wuchereria bancrofti/*immunology/isolation & purification ; }, abstract = {Antifilarial antibody testing has been established as a sensitive and specific method of diagnosing lymphatic filariasis. However, the development of serological responses to specific filarial antigens and their relationship to acquisition of infection is poorly understood. In order to evaluate whether the development of antigen specific antifilarial antibodies precedes microfilaremia and antigenemia, we compared the antibody responses of serum samples collected between 1990 and 1999 from a cohort of 142 Haitian children followed longitudinally. Antigen status was determined using the Og4C3 ELISA and the presence of microfilaremia was detected using microscopy. Antibody responses to Wb123, a Wuchereria bancrofti L3 antigen, were measured using a Luciferase Immunoprecipitation System (LIPS) assay. Antibody responses to Bm14 and Bm33, Brugia malayi antigens and to a major surface protein (WSP) from Wolbachia were analyzed using a multiplex bead assay. Over follow-up, 80 (56%) of the children became antigen-positive and 30 (21%) developed microfilaremia. Detectable antibody responses to Bm14, Bm33, Wb123, and WSP developed in 95%, 100%, 92%, and 29% of children, respectively. With the exception of WSP, the development of antibody responses generally preceded detection of filarial antigen. Our results show that antifilarial antibody responses can serve as an important epidemiological indicator in a sentinel population of young children and thus, may be valuable as tool for surveillance in the context of lymphatic filariasis elimination programs.}, } @article {pmid23236284, year = {2012}, author = {Andrews, ES and Crain, PR and Fu, Y and Howe, DK and Dobson, SL}, title = {Reactive oxygen species production and Brugia pahangi survivorship in Aedes polynesiensis with artificial Wolbachia infection types.}, journal = {PLoS pathogens}, volume = {8}, number = {12}, pages = {e1003075}, pmid = {23236284}, issn = {1553-7374}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*parasitology ; Animals ; Brugia pahangi/*metabolism ; Elephantiasis, Filarial/metabolism/parasitology/prevention & control ; Female ; Oxidative Stress ; Reactive Oxygen Species/*metabolism ; Wolbachia/*metabolism ; }, abstract = {Heterologous transinfection with the endosymbiotic bacterium Wolbachia has been shown previously to induce pathogen interference phenotypes in mosquito hosts. Here we examine an artificially infected strain of Aedes polynesiensis, the primary vector of Wuchereria bancrofti, which is the causative agent of Lymphatic filariasis (LF) throughout much of the South Pacific. Embryonic microinjection was used to transfer the wAlbB infection from Aedes albopictus into an aposymbiotic strain of Ae. polynesiensis. The resulting strain (designated "MTB") experiences a stable artificial infection with high maternal inheritance. Reciprocal crosses of MTB with naturally infected wild-type Ae. polynesiensis demonstrate strong bidirectional incompatibility. Levels of reactive oxygen species (ROS) in the MTB strain differ significantly relative to that of the wild-type, indicating an impaired ability to regulate oxidative stress. Following a challenge with Brugia pahangi, the number of filarial worms achieving the infective stage is significantly reduced in MTB as compared to the naturally infected and aposymbiotic strains. Survivorship of MTB differed significantly from that of the wild-type, with an interactive effect between survivorship and blood feeding. The results demonstrate a direct correlation between decreased ROS levels and decreased survival of adult female Aedes polynesiensis. The results are discussed in relation to the interaction of Wolbachia with ROS production and antioxidant expression, iron homeostasis and the insect immune system. We discuss the potential applied use of the MTB strain for impacting Ae. polynesiensis populations and strategies for reducing LF incidence in the South Pacific.}, } @article {pmid23234643, year = {2012}, author = {Le, PT and Ramulu, HG and Guijarro, L and Paganini, J and Gouret, P and Chabrol, O and Raoult, D and Pontarotti, P}, title = {An automated approach for the identification of horizontal gene transfers from complete genomes reveals the rhizome of Rickettsiales.}, journal = {BMC evolutionary biology}, volume = {12}, number = {}, pages = {243}, pmid = {23234643}, issn = {1471-2148}, mesh = {Alphaproteobacteria/*genetics ; Bayes Theorem ; Electronic Data Processing ; *Evolution, Molecular ; *Gene Transfer, Horizontal ; *Genome, Bacterial ; Genomics/methods ; Phylogeny ; Proteome/analysis ; RNA, Ribosomal, 16S/genetics ; }, abstract = {BACKGROUND: Horizontal gene transfer (HGT) is considered to be a major force driving the evolutionary history of prokaryotes. HGT is widespread in prokaryotes, contributing to the genomic repertoire of prokaryotic organisms, and is particularly apparent in Rickettsiales genomes. Gene gains from both distantly and closely related organisms play crucial roles in the evolution of bacterial genomes. In this work, we focus on genes transferred from distantly related species into Rickettsiales species.

RESULTS: We developed an automated approach for the detection of HGT from other organisms (excluding alphaproteobacteria) into Rickettsiales genomes. Our systematic approach consisted of several specialized features including the application of a parsimony method for inferring phyletic patterns followed by blast filter, automated phylogenetic reconstruction and the application of patterns for HGT detection. We identified 42 instances of HGT in 31 complete Rickettsiales genomes, of which 38 were previously unidentified instances of HGT from Anaplasma, Wolbachia, Candidatus Pelagibacter ubique and Rickettsia genomes. Additionally, putative cases with no phylogenetic support were assigned gene ontology terms. Overall, these transfers could be characterized as "rhizome-like".

CONCLUSIONS: Our analysis provides a comprehensive, systematic approach for the automated detection of HGTs from several complete proteome sequences that can be applied to detect instances of HGT within other genomes of interest.}, } @article {pmid23214526, year = {2012}, author = {Schneider, DM and do Carmo, E and Bar-Yam, Y and de Aguiar, MA}, title = {Robustness against extinction by stochastic sex determination in small populations.}, journal = {Physical review. E, Statistical, nonlinear, and soft matter physics}, volume = {86}, number = {4 Pt 1}, pages = {041104}, doi = {10.1103/PhysRevE.86.041104}, pmid = {23214526}, issn = {1550-2376}, mesh = {Algorithms ; Animals ; Arthropods ; Binomial Distribution ; Female ; Genetics, Population ; Humans ; Male ; Models, Statistical ; Models, Theoretical ; *Population Dynamics ; Probability ; *Sex Determination Processes ; Sex Factors ; Stochastic Processes ; Temperature ; Wolbachia/metabolism ; }, abstract = {Sexually reproducing populations with a small number of individuals may go extinct by stochastic fluctuations in sex determination, causing all their members to become male or female in a generation. In this work we calculate the time to extinction of isolated populations with fixed number N of individuals that are updated according to the Moran birth and death process. At each time step, one individual is randomly selected and replaced by its offspring resulting from mating with another individual of the opposite sex; the offspring can be male or female with equal probability. A set of N time steps is called a generation, the average time it takes for the entire population to be replaced. The number k of females fluctuates in time, similarly to a random walk, and extinction, which is the only asymptotic possibility, occurs when k=0 or k=N. We show that it takes only one generation for an arbitrary initial distribution of males and females to approach the binomial distribution. This distribution, however, is unstable and the population eventually goes extinct in 2(N)/N generations. We also discuss the robustness of these results against bias in the determination of the sex of the offspring, a characteristic promoted by infection by the bacteria Wolbachia in some arthropod species or by temperature in reptiles.}, } @article {pmid23213446, year = {2012}, author = {Landmann, F and Bain, O and Martin, C and Uni, S and Taylor, MJ and Sullivan, W}, title = {Both asymmetric mitotic segregation and cell-to-cell invasion are required for stable germline transmission of Wolbachia in filarial nematodes.}, journal = {Biology open}, volume = {1}, number = {6}, pages = {536-547}, pmid = {23213446}, issn = {2046-6390}, abstract = {Parasitic filarial nematodes that belong to the Onchocercidae family live in mutualism with Wolbachia endosymbionts. We developed whole-mount techniques to follow the segregation patterns of Wolbachia through the somatic and germline lineages of four filarial species. These studies reveal multiple evolutionarily conserved mechanisms that are required for Wolbachia localization to the germline. During the initial embryonic divisions, Wolbachia segregate asymmetrically such that they concentrate in the posteriorly localized P(2) blastomere, a precursor to the adult germline and hypodermal lineages. Surprisingly, in the next division they are excluded from the germline precursor lineage. Rather, they preferentially segregate to the C blastomere, a source of posterior hypodermal cells. Localization to the germline is accomplished by a distinct mechanism in which Wolbachia invade first the somatic gonadal cells close to the ovarian distal tip cell, the nematode stem cell niche, from the hypodermis. This tropism is associated with a cortical F-actin disruption, suggesting an active engulfment. Significantly, germline invasion occurs only in females, explaining the lack of Wolbachia in the male germline. Once in the syncytial environment of the ovaries, Wolbachia rely on the rachis to multiply and disperse into the germ cells. The utilization of cell-to-cell invasion for germline colonization may indicate an ancestral mode of horizontal transfer that preceded the acquisition of the mutualism.}, } @article {pmid23210448, year = {2013}, author = {Taylor, MJ and Voronin, D and Johnston, KL and Ford, L}, title = {Wolbachia filarial interactions.}, journal = {Cellular microbiology}, volume = {15}, number = {4}, pages = {520-526}, doi = {10.1111/cmi.12084}, pmid = {23210448}, issn = {1462-5822}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Filarioidea/*microbiology/physiology ; Survival Analysis ; *Symbiosis ; Wolbachia/drug effects/growth & development/*physiology ; }, abstract = {Wolbachia pipientis is a widespread intracellular bacterial symbiont of arthropods and is common in insects. One of their more exotic and unexpected hosts is the filarial nematodes, notable for the parasites responsible for onchocerciasis (river blindness), lymphatic filariasis (elephantiasis) and dirofilariasis (heartworm). Wolbachia are only present in a subgroup of the filarial nematodes and do not extend to other groups of nematodes either parasitic or free-living. In the medically and veterinary important species that host Wolbachia, the symbiont has become an essential partner to key biological processes in the life of the nematode to the point where antibiotic elimination of the bacteria leads to a potent and effective anti-filarial drug treatment. We review the cellular and molecular basis of Wolbachia filarial interactions and highlight the key processes provided by the endosymbiont upon which the nematodes have become entirely dependent. This dependency is primarily restricted to periods of the lifecycle with heavy metabolic demands including growth and development of larval stages and embryogenesis in the adult female. Also, the longevity of filarial parasites is compromised following depletion of the symbiont, which for the first time has delivered a safe and effective treatment to kill adult parasites with antibiotics.}, } @article {pmid23209869, year = {2012}, author = {Wolbers, M and Kleinschmidt, I and Simmons, CP and Donnelly, CA}, title = {Considerations in the design of clinical trials to test novel entomological approaches to dengue control.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {11}, pages = {e1937}, pmid = {23209869}, issn = {1935-2735}, support = {089276/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Adult ; Child ; Child, Preschool ; *Clinical Trials as Topic ; Dengue/*prevention & control ; Epidemiologic Methods ; Humans ; Mosquito Control/*methods ; *Research Design ; }, } @article {pmid23209856, year = {2012}, author = {Schroeder, JH and Simbi, BH and Ford, L and Cole, SR and Taylor, MJ and Lawson, C and Lawrence, RA}, title = {Live Brugia malayi microfilariae inhibit transendothelial migration of neutrophils and monocytes.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {11}, pages = {e1914}, pmid = {23209856}, issn = {1935-2735}, support = {BB/F011180/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Brugia malayi/*immunology/*pathogenicity ; Cells, Cultured ; Coculture Techniques ; Cytokines/biosynthesis ; Endothelial Cells/*immunology/*parasitology ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique ; Gene Expression Profiling ; Humans ; Microarray Analysis ; Monocytes/*immunology ; Neutrophils/*immunology ; Oligonucleotide Array Sequence Analysis ; Protein Array Analysis ; *Transendothelial and Transepithelial Migration ; }, abstract = {Lymphatic filariasis is a major tropical disease caused by the parasite Brugia malayi. Microfilariae (Mf) circulate in the peripheral blood for 2-3 hours in synchronisation with maximal feeding of the mosquito vector. When absent from the peripheral blood, Mf sequester in the capillaries of the lungs. Mf are therefore in close contact with vascular endothelial cells (EC) and may induce EC immune function and/or wound repair mechanisms such as angiogenesis. In this study, Mf were co-cultured with human umbilical vein EC (HUVEC) or human lung microvascular EC (HLMVEC) and the transendothelial migration of leukocyte subsets was analysed. In addition, the protein and/or mRNA expression of chemokine, cytokine and angiogenic mediators in endothelial cells in the presence of live microfilariae were measured by a combination of cDNA arrays, protein arrays, ELISA and fluorescence antibody tests.Surprisingly, our findings indicate that Mf presence partially blocked transendothelial migration of monocytes and neutrophils, but not lymphocytes. However, Mf exposure did not result in altered vascular EC expression of key mediators of the tethering stage of extravasation, such as ICAM-1, VCAM-1 and various chemokines. To further analyse the immunological function of vascular EC in the presence of Mf, we measured the mRNA and/or protein expression of a number of pro-inflammatory mediators. We found that expression levels of the mediators tested were predominantly unaltered upon B. malayi Mf exposure. In addition, a comparison of angiogenic mediators induced by intact Mf and Wolbachia-depleted Mf revealed that even intact Mf induce the expression of remarkably few angiogenic mediators in vascular EC. Our study suggests that live microfilariae are remarkably inert in their induction and/or activation of vascular cells in their immediate local environment. Overall, this work presents important insights into the immunological function of the vascular endothelium during an infection with B. malayi.}, } @article {pmid23185484, year = {2012}, author = {Osei-Poku, J and Han, C and Mbogo, CM and Jiggins, FM}, title = {Identification of Wolbachia strains in mosquito disease vectors.}, journal = {PloS one}, volume = {7}, number = {11}, pages = {e49922}, pmid = {23185484}, issn = {1932-6203}, support = {//Wellcome Trust/United Kingdom ; 092654//Wellcome Trust/United Kingdom ; WT094664/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Culicidae/microbiology/parasitology ; *Disease Vectors ; Elephantiasis, Filarial/genetics ; Humans ; *Insect Vectors/genetics/microbiology ; *Malaria/genetics/transmission ; Phylogeny ; RNA Viruses/pathogenicity ; *Wolbachia/isolation & purification/pathogenicity ; }, abstract = {Wolbachia bacteria are common endosymbionts of insects, and some strains are known to protect their hosts against RNA viruses and other parasites. This has led to the suggestion that releasing Wolbachia-infected mosquitoes could prevent the transmission of arboviruses and other human parasites. We have identified Wolbachia in Kenyan populations of the yellow fever vector Aedes bromeliae and its relative Aedes metallicus, and in Mansonia uniformis and Mansonia africana, which are vectors of lymphatic filariasis. These Wolbachia strains cluster together on the bacterial phylogeny, and belong to bacterial clades that have recombined with other unrelated strains. These new Wolbachia strains may be affecting disease transmission rates of infected mosquito species, and could be transferred into other mosquito vectors as part of control programs.}, } @article {pmid23183308, year = {2013}, author = {Martin, OY and Puniamoorthy, N and Gubler, A and Wimmer, C and Bernasconi, MV}, title = {Infections with Wolbachia, Spiroplasma, and Rickettsia in the Dolichopodidae and other Empidoidea.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {13}, number = {}, pages = {317-330}, doi = {10.1016/j.meegid.2012.11.005}, pmid = {23183308}, issn = {1567-7257}, mesh = {Animals ; DNA, Bacterial ; Diptera/*microbiology ; Male ; Molecular Sequence Data ; Phylogeny ; Rickettsia/classification/*genetics ; Spiroplasma/classification/*genetics ; Symbiosis ; Wolbachia/classification/*genetics ; }, abstract = {Vertically transmitted reproductive parasites are both extraordinarily widespread and diverse in their effects on their invertebrate hosts. In addition to causing skewed population sex ratios via male-killing or feminization, such bacteria can further cause cytoplasmic incompatibility or parthenogenesis. Previous surveys show that the microbes Wolbachia and Spiroplasma are common in some dipteran families, e.g. Drosophilidae or Scathophagidae, and are known to be heritable symbionts and affect reproduction in the Diptera. However, little is known of Rickettsia infections and detailed surveys targeting other Dipteran families are lacking. Here 329 samples of 247 species of Diptera belonging to the Dolichopodidae, Empididae, and Hybotidae (superfamily Empidoidea) are surveyed for the presence of the endosymbionts Wolbachia, Spiroplasma, and Rickettsia. The superfamily Empidoidea contains numerous species, which have been the targets of intense research concerning reproductive traits involved in sexual selection. 151 of the species (i.e. ca. 61%) screened here, including species from key genera such as Dolichopus, Poecilobothrus or Empis, harboured one or more symbionts. Reproductive parasites are thus also common in the Empidoidae, yet effects on hosts remain unclear. Potential endosymbiont-host interactions in this group would hence be worthy of further investigation.}, } @article {pmid23182274, year = {2012}, author = {Subramanian, G and Sekeyova, Z and Raoult, D and Mediannikov, O}, title = {Multiple tick-associated bacteria in Ixodes ricinus from Slovakia.}, journal = {Ticks and tick-borne diseases}, volume = {3}, number = {5-6}, pages = {406-410}, doi = {10.1016/j.ttbdis.2012.10.001}, pmid = {23182274}, issn = {1877-9603}, mesh = {Animals ; Bacteria/*classification/genetics/*isolation & purification ; Ixodes/*microbiology ; Polymerase Chain Reaction ; Slovakia ; Trees ; }, abstract = {Eighty Ixodes ricinus ticks collected in the Rovinka forest in Slovakia were tested by real-time and routine single-step PCR for the presence of different pathogenic and endosymbiotic bacteria. No evidence of Coxiella burnetii, Diplorickettsia massiliensis, or Bartonella spp. was found. However, we identified, as the pathogenic bacteria, Anaplasma phagocytophilum in 8%, Rickettsia helvetica in 11%, Borrelia afzelii in 9%, Bo. garinii in 8%, Bo. burgdorferi sensu stricto in 5%, Bo. valaisiana in 3%, and Bo. miyamotoi in 1% of ticks. Other bacteria with unknown or suspected pathogenicity for humans were also identified: Arsenophonus nasoniae in 25%, Spiroplasma ixodetis in 3%, Candidatus Midichloria mitochondrii in 4%, Wolbachia aff. pipientis in 14%, and Ehrlichia muris in 3% of ticks. A. nasoniae was found almost exclusively in nymphs, and E. muris was only found in adult ticks. In total, 65% of ticks contained at least one species of bacteria, and 16% contained 2 or more species.}, } @article {pmid23171418, year = {2013}, author = {Moretti, R and Calvitti, M}, title = {Male mating performance and cytoplasmic incompatibility in a wPip Wolbachia trans-infected line of Aedes albopictus (Stegomyia albopicta).}, journal = {Medical and veterinary entomology}, volume = {27}, number = {4}, pages = {377-386}, doi = {10.1111/j.1365-2915.2012.01061.x}, pmid = {23171418}, issn = {1365-2915}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Female ; Male ; Mosquito Control ; Pest Control, Biological/methods ; Reproduction/physiology ; Sexual Behavior, Animal ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis Hertig (Rickettsiales: Rickettsiaceae) is a maternally inherited endosymbiont of a large number of insects and other arthropods that induces various effects on host reproductive biology. Among these, cytoplasmic incompatibility (CI) is a form of sterility induced in eggs produced by mating between infected males and females uninfected or infected by an incompatible Wolbachia strain. This phenomenon has been proposed as a potential way to produce functionally sterile males to be used in genetic control programmes. In this paper, we report on experiments carried out to evaluate the mating performances of males of an Aedes albopictus (Stegomyia albopicta) (Diptera: Culicidae) line (ARwP), harbouring a new Wolbachia infection [the wPip strain from Culex pipiens Linnaeus (Diptera: Culicidae)], in comparison with naturally infected males (SR line). ARwP males did not differ from SR males with regard to insemination capacity. Mating competitiveness did not differ significantly between lines in either laboratory or greenhouse conditions. Moreover, crosses with SR females were characterized by a 100% CI regardless of ARwP male age. All of these findings suggest that ARwP males may represent a very efficient tool for control programmes against Ae. albopictus based on the release of functionally sterile males.}, } @article {pmid23166845, year = {2012}, author = {O'Connor, L and Plichart, C and Sang, AC and Brelsfoard, CL and Bossin, HC and Dobson, SL}, title = {Open release of male mosquitoes infected with a wolbachia biopesticide: field performance and infection containment.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {11}, pages = {e1797}, pmid = {23166845}, issn = {1935-2735}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Female ; Male ; Mice ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: Lymphatic filariasis (LF) is a globally significant disease, with 1.3 billion persons in 83 countries at risk. A coordinated effort of administering annual macrofilaricidal prophylactics to the entire at-risk population has succeeded in impacting and eliminating LF transmission in multiple regions. However, some areas in the South Pacific are predicted to persist as transmission sites, due in part to the biology of the mosquito vector, which has led to a call for additional tools to augment drug treatments. Autocidal strategies against mosquitoes are resurging in the effort against invasive mosquitoes and vector borne disease, with examples that include field trials of genetically modified mosquitoes and Wolbachia population replacement. However, critical questions must be addressed in anticipation of full field trials, including assessments of field competitiveness of transfected males and the risk of unintended population replacement.

We report the outcome of field experiments testing a strategy that employs Wolbachia as a biopesticide. The strategy is based upon Wolbachia-induced conditional sterility, known as cytoplasmic incompatibility, and the repeated release of incompatible males to suppress a population. A criticism of the Wolbachia biopesticide approach is that unintended female release or horizontal Wolbachia transmission can result in population replacement instead of suppression. We present the outcome of laboratory and field experiments assessing the competitiveness of transfected males and their ability to transmit Wolbachia via horizontal transmission.

CONCLUSIONS/SIGNIFICANCE: The results demonstrate that Wolbachia-transfected Aedes polynesiensis males are competitive under field conditions during a thirty-week open release period, as indicated by mark, release, recapture and brood-hatch failure among females at the release site. Experiments demonstrate the males to be 'dead end hosts' for Wolbachia and that methods were adequate to prevent population replacement at the field site. The findings encourage the continued development and extension of a Wolbachia autocidal approach to additional medically important mosquito species.}, } @article {pmid23166822, year = {2012}, author = {Saha, S and Hunter, WB and Reese, J and Morgan, JK and Marutani-Hert, M and Huang, H and Lindeberg, M}, title = {Survey of endosymbionts in the Diaphorina citri metagenome and assembly of a Wolbachia wDi draft genome.}, journal = {PloS one}, volume = {7}, number = {11}, pages = {e50067}, pmid = {23166822}, issn = {1932-6203}, mesh = {Animals ; Base Sequence ; Demography ; Genome, Bacterial/*genetics ; Hemiptera/genetics/*microbiology ; Metagenome/*genetics ; Molecular Sequence Annotation ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA ; Symbiosis/*genetics ; Wolbachia/*genetics ; }, abstract = {Diaphorina citri (Hemiptera: Psyllidae), the Asian citrus psyllid, is the insect vector of Ca. Liberibacter asiaticus, the causal agent of citrus greening disease. Sequencing of the D. citri metagenome has been initiated to gain better understanding of the biology of this organism and the potential roles of its bacterial endosymbionts. To corroborate candidate endosymbionts previously identified by rDNA amplification, raw reads from the D. citri metagenome sequence were mapped to reference genome sequences. Results of the read mapping provided the most support for Wolbachia and an enteric bacterium most similar to Salmonella. Wolbachia-derived reads were extracted using the complete genome sequences for four Wolbachia strains. Reads were assembled into a draft genome sequence, and the annotation assessed for the presence of features potentially involved in host interaction. Genome alignment with the complete sequences reveals membership of Wolbachia wDi in supergroup B, further supported by phylogenetic analysis of FtsZ. FtsZ and Wsp phylogenies additionally indicate that the Wolbachia strain in the Florida D. citri isolate falls into a sub-clade of supergroup B, distinct from Wolbachia present in Chinese D. citri isolates, supporting the hypothesis that the D. citri introduced into Florida did not originate from China.}, } @article {pmid23166816, year = {2012}, author = {Osei-Amo, S and Hussain, M and O'Neill, SL and Asgari, S}, title = {Wolbachia-induced aae-miR-12 miRNA negatively regulates the expression of MCT1 and MCM6 genes in Wolbachia-infected mosquito cell line.}, journal = {PloS one}, volume = {7}, number = {11}, pages = {e50049}, pmid = {23166816}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Animals ; Cell Cycle Proteins/metabolism ; Computational Biology ; Gene Expression Regulation/*genetics ; MicroRNAs/genetics/*metabolism ; Monocarboxylic Acid Transporters/metabolism ; Real-Time Polymerase Chain Reaction ; Sf9 Cells ; Spodoptera ; Symporters/metabolism ; Wolbachia/growth & development/*metabolism ; }, abstract = {BACKGROUND: Best recognized for its role in manipulating host reproduction, the parasitic gram-negative Wolbachia pipientis is known to colonize a wide range of invertebrates. The endosymbiotic bacterium has recently been shown to cause a life-shortening effect as well as inhibiting replication of arboviruses in Aedes aegypti; although the molecular mechanisms behind these effects are largely unknown. MicroRNAs (miRNAs) have been determined to have a wide range of roles in regulating gene expression in eukaryotes. A recent study showed that several A. aegypti mosquito miRNAs are differentially expressed when infected with Wolbachia.

Based on the prior knowledge that one of these miRNAs, aae-miR-12, is differentially expressed in mosquitoes infected with Wolbachia, we aimed to determine any significance of this mediation. We also set out to characterize the target genes of this miRNA in the A. aegpyti genome. Bioinformatic approaches predicted a list of potential target genes and subsequent functional analyses confirmed that two of these, DNA replication licensing (MCM6) and monocarboxylate transporter (MCT1), are under the regulative control of aae-miR-12. We also demonstrated that aae-miR-12 is critical in the persistence of Wolbachia in the host cell.

CONCLUSIONS/SIGNIFICANCE: Our study has identified two target genes of aae-miR-12, a differentially expressed mosquito miRNA in Wolbachia-infected cells, and determined that the miRNA affects Wolbachia density in the host cells.}, } @article {pmid23154238, year = {2012}, author = {Short, S and Yang, G and Kille, P and Ford, AT}, title = {A widespread and distinctive form of amphipod intersexuality not induced by known feminising parasites.}, journal = {Sexual development : genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation}, volume = {6}, number = {6}, pages = {320-324}, doi = {10.1159/000343779}, pmid = {23154238}, issn = {1661-5433}, support = {G0900740/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Amphipoda/*parasitology/*physiology ; Animals ; Disorders of Sex Development/parasitology/*veterinary ; Feminization/parasitology/veterinary ; Male ; Microsporidia ; Phenotype ; Seasons ; Testis ; Wolbachia ; }, abstract = {Intersexuality occurs in a diverse range of animals, and its study offers insights into basic reproductive biology. Investigations in amphipods suggest intersexuality results from incomplete feminisation caused by sex-distorting parasites. It has also been noted that 2 intersex phenotypes occur in males of the amphipod Echinogammarus marinus, an external phenotype, in which males possess rudimentary brood plates, and an internal phenotype, in which only an ovotestis is present. This study examines the relationship between these phenotypes and finds their prevalences are independent. In addition, a cross-species microarray reveals the testicular transcriptomes of the intersex phenotypes are distinct from that of normal males and, most crucially, each other. Furthermore, the internal intersex phenotype, unlike the external phenotype, shows no correlation with infection by known feminising parasites. These findings suggest the male intersex phenotypes should not be considered stages on a single spectrum of intersexuality. Rather, they support the hypothesis that internal and external intersexuality are divergent phenotypes with separate causal mechanisms and point to the existence of a distinct and geographically widespread form of amphipod intersexuality caused by an unknown factor.}, } @article {pmid23152317, year = {2012}, author = {Morais, SA and Almeida, Fd and Suesdek, L and Marrelli, MT}, title = {Low genetic diversity in Wolbachia-Infected Culex quinquefasciatus (Diptera: Culicidae) from Brazil and Argentina.}, journal = {Revista do Instituto de Medicina Tropical de Sao Paulo}, volume = {54}, number = {6}, pages = {325-329}, doi = {10.1590/s0036-46652012000600007}, pmid = {23152317}, issn = {1678-9946}, mesh = {Animals ; Argentina ; Brazil ; Culex/*genetics/*microbiology ; Electron Transport Complex IV/*genetics ; Genes, Insect/genetics ; Genes, Mitochondrial/genetics ; Genetic Variation/*genetics ; NADH Dehydrogenase/*genetics ; *Wolbachia ; }, abstract = {Culex quinquefasciatus is a vector of human pathogens, including filarial nematodes and several viruses. Although its epidemiological relevance is known to vary across geographical regions, an understanding of its population genetic structure is still incipient. In light of this, we evaluated the genetic diversity of Cx. quinquefasciatus and Cx. pipiens x Cx. quinquefasciatus hybrids collected from nine localities in Brazil and one site in Argentina. We used mitochondrial genes cox1 and nd4, along with the coxA and wsp genes of the maternally-inherited Wolbachia endosymbiont. The nd4 fragment was invariant between samples, whilst cox1 exhibited four haplotypes that separated two types of Cx. quinquefasciatus, one clustered in southern Brazil. Low sequence diversity was generally observed, being discussed. Both Brazilian and Argentinian mosquitoes were infected with a single Wolbachia strain. As reported in previous studies with these populations, cox1 and nd4 diversity is not congruent with the population structure revealed by nuclear markers or alar morphology. Future Cx. quinquefasciatus research should, if possible, evaluate mtDNA diversity in light of other markers.}, } @article {pmid23151511, year = {2012}, author = {Bronkhorst, AW and van Cleef, KW and Vodovar, N and Ince, IA and Blanc, H and Vlak, JM and Saleh, MC and van Rij, RP}, title = {The DNA virus Invertebrate iridescent virus 6 is a target of the Drosophila RNAi machinery.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {109}, number = {51}, pages = {E3604-13}, pmid = {23151511}, issn = {1091-6490}, mesh = {Animals ; Antiviral Agents/chemistry ; DNA Viruses/*genetics ; Drosophila Proteins/metabolism ; Drosophila melanogaster ; *Gene Expression Regulation, Viral ; Gene Silencing ; Genome, Viral ; Kinetics ; Models, Genetic ; Mutation ; Polymerase Chain Reaction ; *RNA Interference ; RNA, Small Interfering/metabolism ; Sequence Analysis, DNA ; Wolbachia/metabolism ; }, abstract = {RNA viruses in insects are targets of an RNA interference (RNAi)-based antiviral immune response, in which viral replication intermediates or viral dsRNA genomes are processed by Dicer-2 (Dcr-2) into viral small interfering RNAs (vsiRNAs). Whether dsDNA virus infections are controlled by the RNAi pathway remains to be determined. Here, we analyzed the role of RNAi in DNA virus infection using Drosophila melanogaster infected with Invertebrate iridescent virus 6 (IIV-6) as a model. We show that Dcr-2 and Argonaute-2 mutant flies are more sensitive to virus infection, suggesting that vsiRNAs contribute to the control of DNA virus infection. Indeed, small RNA sequencing of IIV-6-infected WT and RNAi mutant flies identified abundant vsiRNAs that were produced in a Dcr-2-dependent manner. We observed a highly uneven distribution with strong clustering of vsiRNAs to small defined regions (hotspots) and modest coverage at other regions (coldspots). vsiRNAs mapped in similar proportions to both strands of the viral genome, suggesting that long dsRNA derived from convergent overlapping transcripts serves as a substrate for Dcr-2. In agreement, strand-specific RT-PCR and Northern blot analyses indicated that antisense transcripts are produced during infection. Moreover, we show that vsiRNAs are functional in silencing reporter constructs carrying fragments of the IIV-6 genome. Together, our data indicate that RNAi provides antiviral defense against dsDNA viruses in animals. Thus, RNAi is the predominant antiviral defense mechanism in insects that provides protection against all major classes of viruses.}, } @article {pmid23149015, year = {2013}, author = {Endersby, NM and Hoffmann, AA}, title = {Effect of Wolbachia on insecticide susceptibility in lines of Aedes aegypti.}, journal = {Bulletin of entomological research}, volume = {103}, number = {3}, pages = {269-277}, doi = {10.1017/S0007485312000673}, pmid = {23149015}, issn = {1475-2670}, mesh = {Aedes/*drug effects/*microbiology ; Animals ; Bacillus thuringiensis/pathogenicity ; Dengue/prevention & control ; Insecticide Resistance/*physiology ; Insecticides/*toxicity ; Methoprene/toxicity ; Pest Control, Biological/*methods ; Pyrethrins/toxicity ; Temefos/toxicity ; Wolbachia/*physiology ; }, abstract = {Two stable infections of Wolbachia pipientis, wMelPop and wMel, now established in Aedes aegypti, are being used in a biocontrol program to suppress the transmission of dengue. Any effects of Wolbachia infection on insecticide resistance of mosquitoes may undermine the success of this program. Bioassays of Ae. aegypti were conducted to test for differences in response to insecticides between Wolbachia infected (wMelPop, wMel) and uninfected lines. Insecticides screened were bifenthrin, the pyrethroid commonly used for adult knockdown, as well as larvicides: Bacillus thuringiensis var. israelensis, the organophosphate, temephos and the insect growth regulator, s-methoprene. While differences in response between lines were detected for some insecticides, no obvious or consistent effects related to presence of Wolbachia infection were observed. Spreading Wolbachia infections are, therefore, unlikely to affect the efficacy of traditional chemical control of mosquito outbreaks.}, } @article {pmid23146564, year = {2012}, author = {Calvitti, M and Moretti, R and Skidmore, AR and Dobson, SL}, title = {Wolbachia strain wPip yields a pattern of cytoplasmic incompatibility enhancing a Wolbachia-based suppression strategy against the disease vector Aedes albopictus.}, journal = {Parasites & vectors}, volume = {5}, number = {}, pages = {254}, pmid = {23146564}, issn = {1756-3305}, mesh = {Aedes/*microbiology/physiology ; Age Factors ; Animals ; Crosses, Genetic ; Cytoplasm ; Female ; Fertility ; Host-Pathogen Interactions ; Humans ; Insect Vectors/*microbiology/physiology ; Male ; Ovum ; Pest Control, Biological/methods ; Reproduction ; Sex Factors ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {BACKGROUND: Cytoplasmic incompatibility (CI) is induced in nature by Wolbachia bacteria, resulting in conditional male sterility. Previous research demonstrated that the two Wolbachia strains (wAlbA and wAlbB) that naturally co-infect the disease vector mosquito Aedes albopictus (Asian tiger mosquito) can be replaced with the wPip Wolbachia strain from Culex pipiens. Since Wolbachia-based vector control strategies depend upon the strength and consistency of CI, a greater understanding is needed on the CI relationships between wPip, wAlbA and wAlbB Wolbachia in Ae. albopictus.

METHODS: This work consisted of a collaborative series of crosses carried out in Italy and in US to study the CI relationships between the "wPip" infected Ae. albopictus strain (ARwP) and the superinfected SR strain. The Ae. albopictus strains used in Italian tests are the wPip infected ARwP strain (ARwPIT), the superinfected SR strain and the aposymbiotic AR strain. To understand the observed pattern of CI, crossing experiments carried out in USA focused on the study of the CI relationships between ARwP (ARwPUS) and artificially-generated single infected lines, in specific HTA and HTB, harbouring only wAlbA and wAlbB Wolbachia respectively.

RESULTS: The paper reports an unusual pattern of CI observed in crossing experiments between ARwP and SR lines. Specifically, ARwP males are able to induce full sterility in wild type females throughout most of their lifetime, while crosses between SR males and ARwP females become partially fertile with male aging. We demonstrated that the observed decrease in CI penetrance with SR male age, is related to the previously described decrease in Wolbachia density, in particular of the wAlbA strain, occurring in aged superinfected males.

CONCLUSIONS: The results here reported support the use of the ARwP Ae. albopictus line as source of "ready-made sterile males", as an alternative to gamma radiation sterilized males, for autocidal suppression strategies against the Asian tiger mosquito. In addition, the age dependent CI weakening observed in the crosses between SR males and ARwP females simplifies the downstream efforts to preserve the genetic variability within the laboratory ARwP colonies, to date based on the antibiotic treatment of wild captured superinfected mosquitoes, also reducing the costs.}, } @article {pmid23145008, year = {2012}, author = {Yang, CY and Xiao, JH and Niu, LM and Ma, GC and Cook, JM and Bian, SN and Fu, YG and Huang, DW}, title = {Chaos of Wolbachia sequences inside the compact fig syconia of Ficus benjamina (Ficus: moraceae).}, journal = {PloS one}, volume = {7}, number = {11}, pages = {e48882}, pmid = {23145008}, issn = {1932-6203}, mesh = {Animals ; Bacterial Outer Membrane Proteins/chemistry/genetics ; Female ; Ficus ; Genotype ; Male ; Phylogeny ; Population Dynamics ; Sequence Analysis, DNA ; Symbiosis ; Wasps/*microbiology ; Wolbachia/*genetics ; }, abstract = {Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium (inflorescence) habitat for the lives of a complex assemblage of Chalcidoid insects. These diverse fig wasp species have intimate ecological relationships within the closed world of the fig syconia. Previous surveys of Wolbachia, maternally inherited endosymbiotic bacteria that infect vast numbers of arthropod hosts, showed that fig wasps have some of the highest known incidences of Wolbachia amongst all insects. We ask whether the evolutionary patterns of Wolbachia sequences in this closed syconium community are different from those in the outside world. In the present study, we sampled all 17 fig wasp species living on Ficus benjamina, covering 4 families, 6 subfamilies, and 8 genera of wasps. We made a thorough survey of Wolbachia infection patterns and studied evolutionary patterns in wsp (Wolbachia Surface Protein) sequences. We find evidence for high infection incidences, frequent recombination between Wolbachia strains, and considerable horizontal transfer, suggesting rapid evolution of Wolbachia sequences within the syconium community. Though the fig wasps have relatively limited contact with outside world, Wolbachia may be introduced to the syconium community via horizontal transmission by fig wasps species that have winged males and visit the syconia earlier.}, } @article {pmid23144739, year = {2012}, author = {Xue, X and Li, SJ and Ahmed, MZ and De Barro, PJ and Ren, SX and Qiu, BL}, title = {Inactivation of Wolbachia reveals its biological roles in whitefly host.}, journal = {PloS one}, volume = {7}, number = {10}, pages = {e48148}, pmid = {23144739}, issn = {1932-6203}, mesh = {Animals ; Antibiotics, Antitubercular/pharmacology ; Body Size/drug effects ; Female ; Gene Expression/drug effects ; Genes, Bacterial/genetics ; Hemiptera/genetics/*microbiology/parasitology ; Host-Pathogen Interactions/drug effects ; In Situ Hybridization, Fluorescence ; Longevity/drug effects ; Male ; Nymph/genetics/microbiology/parasitology ; Parasite Egg Count ; RNA, Ribosomal, 16S/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Rifampin/pharmacology ; Sex Ratio ; Symbiosis/drug effects/genetics/*physiology ; Wasps/physiology ; Wolbachia/drug effects/genetics/*physiology ; }, abstract = {BACKGROUND: The whitefly Bemisia tabaci is cryptic species complex composed of numerous species. Individual species from the complex harbor a diversity of bacterial endosymbionts including Wolbachia. However, while Wolbachia is known to have a number of different roles, its role in B. tabaci is unclear. Here, the antibiotic rifampicin is used to selectively eliminate Wolbachia from B. tabaci so as to enable its roles in whitefly development and reproduction to be explored. The indirect effects of Wolbachia elimination on the biology of Encarsia bimaculata, a dominant parasitoid of B. tabaci in South China, were also investigated.

METHODOLOGY/PRINCIPAL FINDING: qRT-PCR and FISH were used to show that after 48 h exposure to 1.0 mg/ml rifampicin, Wolbachia was completely inactivated from B. tabaci Mediterranean (MED) without any significant impact on either the primary symbiont, Portiera aleyrodidarum or any of the other secondary endosymbionts present. For B. tabaci MED, Wolbachia was shown to be associated with decreased juvenile development time, increased likelihood that nymphs completed development, increased adult life span and increased percentage of female progeny. Inactivation was associated with a significant decrease in the body size of the 4(th) instar which leads us to speculate as to whether Wolbachia may have a nutrient supplementation role. The reduction in nymph body size has consequences for its parasitoid, E. bimaculata. The elimination of Wolbachia lead to a marked increase in the proportion of parasitoid eggs that completed their development, but the reduced size of the whitefly host was also associated with a significant reduction in the size of the emerging parasitoid adult and this was in turn associated with a marked reduction in adult parasitoid longevity.

CONCLUSIONS/SIGNIFICANCE: Wolbachia increases the fitness of the whitefly host and provides some protection against parasitization. These observations add to our understanding of the roles played by bacterial endosymbionts.}, } @article {pmid23133693, year = {2012}, author = {van den Hurk, AF and Hall-Mendelin, S and Pyke, AT and Frentiu, FD and McElroy, K and Day, A and Higgs, S and O'Neill, SL}, title = {Impact of Wolbachia on infection with chikungunya and yellow fever viruses in the mosquito vector Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {11}, pages = {e1892}, pmid = {23133693}, issn = {1935-2735}, support = {UC7 AI070083/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; *Antibiosis ; Chikungunya virus/*isolation & purification ; *Disease Vectors ; Wolbachia/*physiology ; Yellow fever virus/*isolation & purification ; }, abstract = {Incidence of disease due to dengue (DENV), chikungunya (CHIKV) and yellow fever (YFV) viruses is increasing in many parts of the world. The viruses are primarily transmitted by Aedes aegypti, a highly domesticated mosquito species that is notoriously difficult to control. When transinfected into Ae. aegypti, the intracellular bacterium Wolbachia has recently been shown to inhibit replication of DENVs, CHIKV, malaria parasites and filarial nematodes, providing a potentially powerful biocontrol strategy for human pathogens. Because the extent of pathogen reduction can be influenced by the strain of bacterium, we examined whether the wMel strain of Wolbachia influenced CHIKV and YFV infection in Ae. aegypti. Following exposure to viremic blood meals, CHIKV infection and dissemination rates were significantly reduced in mosquitoes with the wMel strain of Wolbachia compared to Wolbachia-uninfected controls. However, similar rates of infection and dissemination were observed in wMel infected and non-infected Ae. aegypti when intrathoracic inoculation was used to deliver virus. YFV infection, dissemination and replication were similar in wMel-infected and control mosquitoes following intrathoracic inoculations. In contrast, mosquitoes with the wMelPop strain of Wolbachia showed at least a 10(4) times reduction in YFV RNA copies compared to controls. The extent of reduction in virus infection depended on Wolbachia strain, titer and strain of the virus, and mode of exposure. Although originally proposed for dengue biocontrol, our results indicate a Wolbachia-based strategy also holds considerable promise for YFV and CHIKV suppression.}, } @article {pmid23133394, year = {2012}, author = {Penz, T and Schmitz-Esser, S and Kelly, SE and Cass, BN and Müller, A and Woyke, T and Malfatti, SA and Hunter, MS and Horn, M}, title = {Comparative genomics suggests an independent origin of cytoplasmic incompatibility in Cardinium hertigii.}, journal = {PLoS genetics}, volume = {8}, number = {10}, pages = {e1003012}, pmid = {23133394}, issn = {1553-7404}, support = {P 22703/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Arthropods/microbiology ; Bacterial Proteins/genetics/metabolism ; Bacteroidetes/*genetics/metabolism ; Biological Evolution ; Biosynthetic Pathways ; Cytoplasm/metabolism ; Gene Order ; *Genome, Bacterial ; *Genomics ; Host-Pathogen Interactions ; Symbiosis/genetics ; }, abstract = {Terrestrial arthropods are commonly infected with maternally inherited bacterial symbionts that cause cytoplasmic incompatibility (CI). In CI, the outcome of crosses between symbiont-infected males and uninfected females is reproductive failure, increasing the relative fitness of infected females and leading to spread of the symbiont in the host population. CI symbionts have profound impacts on host genetic structure and ecology and may lead to speciation and the rapid evolution of sex determination systems. Cardinium hertigii, a member of the Bacteroidetes and symbiont of the parasitic wasp Encarsia pergandiella, is the only known bacterium other than the Alphaproteobacteria Wolbachia to cause CI. Here we report the genome sequence of Cardinium hertigii cEper1. Comparison with the genomes of CI-inducing Wolbachia pipientis strains wMel, wRi, and wPip provides a unique opportunity to pinpoint shared proteins mediating host cell interaction, including some candidate proteins for CI that have not previously been investigated. The genome of Cardinium lacks all major biosynthetic pathways but harbors a complete biotin biosynthesis pathway, suggesting a potential role for Cardinium in host nutrition. Cardinium lacks known protein secretion systems but encodes a putative phage-derived secretion system distantly related to the antifeeding prophage of the entomopathogen Serratia entomophila. Lastly, while Cardinium and Wolbachia genomes show only a functional overlap of proteins, they show no evidence of laterally transferred elements that would suggest common ancestry of CI in both lineages. Instead, comparative genomics suggests an independent evolution of CI in Cardinium and Wolbachia and provides a novel context for understanding the mechanistic basis of CI.}, } @article {pmid23123345, year = {2013}, author = {Gayen, P and Nayak, A and Saini, P and Mukherjee, N and Maitra, S and Sarkar, P and Sinha Babu, SP}, title = {A double-blind controlled field trial of doxycycline and albendazole in combination for the treatment of bancroftian filariasis in India.}, journal = {Acta tropica}, volume = {125}, number = {2}, pages = {150-156}, doi = {10.1016/j.actatropica.2012.10.011}, pmid = {23123345}, issn = {1873-6254}, mesh = {Adolescent ; Adult ; Aged ; Albendazole/administration & dosage/*therapeutic use ; Animals ; Anthelmintics/administration & dosage/therapeutic use ; Anti-Bacterial Agents/administration & dosage/therapeutic use ; Bacterial Load ; Double-Blind Method ; Doxycycline/administration & dosage/*therapeutic use ; Drug Administration Schedule ; Drug Therapy, Combination ; Elephantiasis, Filarial/blood/*drug therapy/parasitology ; Female ; Humans ; India ; Logistic Models ; Male ; Middle Aged ; Parasite Load ; Time Factors ; Wolbachia/*drug effects ; Wuchereria bancrofti/*drug effects/*microbiology ; Young Adult ; }, abstract = {In a placebo controlled field trial, the effects of doxycycline (200mg/day) for 23 days followed by doxycycline (200mg/day) in combination with albendazole (ABZ) (400mg/day) for 7 days on depletion of Wolbachia endobacteria from Wuchereria bancrofti and microfilaricidal activity were studied in 68 patients (34 males and 34 females) from West Bengal, India. The drugs in combination (i.e., doxycycline+ABZ) provided the best efficacy by totally eliminating the circulating microfilaria (mf) (in 42% cases) on day 365 with (99.8%, P<0.05) suppression even on day 365 post-treatment compared to both exclusive doxycycline (69%, P<0.05) and ABZ (89%, P<0.05) groups. Thus, our results have established that a 30-day course of doxycycline in combination with a 7-day course of ABZ is sufficient to ensure long-term reduction in mf level by depleting Wolbachia from worm tissues. Doxycycline combined with ABZ led to a greater reduction in mf density in blood at 4 months (post-treatment) in comparison to doxycycline or ABZ alone. There were significant differences between the three treatments after 12 months (post-treatment). Further, the impact of a 7-day regimen of ABZ was surprisingly good in reducing mf compared to doxycycline-alone group. Adverse reactions were mild. A 30-day course of doxycycline and ABZ in combination is a safe and well-tolerated treatment for lymphatic filariasis with significant activity against microfilaremia.}, } @article {pmid23115298, year = {2013}, author = {Hussain, M and Lu, G and Torres, S and Edmonds, JH and Kay, BH and Khromykh, AA and Asgari, S}, title = {Effect of Wolbachia on replication of West Nile virus in a mosquito cell line and adult mosquitoes.}, journal = {Journal of virology}, volume = {87}, number = {2}, pages = {851-858}, pmid = {23115298}, issn = {1098-5514}, mesh = {Aedes/*virology ; Animals ; *Antibiosis ; Cell Line ; *Virus Replication ; West Nile virus/growth & development/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia as an endosymbiont is widespread in insects and other arthropods and is best known for reproductive manipulations of the host. Recently, it has been shown that wMelpop and wMel strains of Wolbachia inhibit the replication of several RNA viruses, including dengue virus, and other vector-borne pathogens (e.g., Plasmodium and filarial nematodes) in mosquitoes, providing an alternative approach to limit the transmission of vector-borne pathogens. In this study, we tested the effect of Wolbachia on the replication of West Nile Virus (WNV). Surprisingly, accumulation of the genomic RNA of WNV for all three strains of WNV tested (New York 99, Kunjin, and New South Wales) was enhanced in Wolbachia-infected Aedes aegypti cells (Aag2). However, the amount of secreted virus was significantly reduced in the presence of Wolbachia. Intrathoracic injections showed that replication of WNV in A. aegypti mosquitoes infected with wMel strain of Wolbachia was not inhibited, whereas wMelPop strain of Wolbachia significantly reduced the replication of WNV in mosquitoes. Further, when wMelPop mosquitoes were orally fed with WNV, virus infection, transmission, and dissemination rates were very low in Wolbachia-free mosquitoes and were completely inhibited in the presence of Wolbachia. The results suggest that (i) despite the enhancement of viral genomic RNA replication in the Wolbachia-infected cell line the production of secreted virus was significantly inhibited, (ii) the antiviral effect in intrathoracically infected mosquitoes depends on the strain of Wolbachia, and (iii) replication of the virus in orally fed mosquitoes was completely inhibited in wMelPop strain of Wolbachia.}, } @article {pmid23113940, year = {2013}, author = {Khoo, CC and Venard, CM and Fu, Y and Mercer, DR and Dobson, SL}, title = {Infection, growth and maintenance of Wolbachia pipientis in clonal and non-clonal Aedes albopictus cell cultures.}, journal = {Bulletin of entomological research}, volume = {103}, number = {3}, pages = {251-260}, doi = {10.1017/S0007485312000648}, pmid = {23113940}, issn = {1475-2670}, support = {R01-AI051533/AI/NIAID NIH HHS/United States ; R01-AI067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*cytology/*microbiology ; Animals ; Bacterial Proteins/genetics ; Cell Culture Techniques/methods ; Cell Line ; DNA Primers/genetics ; Drosophila/*microbiology ; In Situ Hybridization, Fluorescence ; Indoles ; Polymerase Chain Reaction ; Wolbachia/*genetics/*growth & development ; }, abstract = {Insect cell lines provide useful in vitro models for studying biological systems, including interactions between mosquitoes and obligate intracellular endosymbionts such as Wolbachia pipientis. The Aedes albopictus Aa23 cell line was the first cell line developed to allow examination of Wolbachia infections. However, Wolbachia studies using Aa23 can be complicated by the presence of different cell types in the cell line and the substantial temporal variation in infection level. Two approaches were examined to ameliorate infection variability. In the first approach, multiple Aa23 passaging regimes were tested for an effect on infection variability. Fluorescence in situ hybridization (FISH) staining was used to characterize Wolbachia infection level over time. The results demonstrate an impact of passaging method on Wolbachia infection level, with some methods resulting in loss of infection. None of the passaging methods succeeded in effectively mitigating infection level variation. In a second approach, the clonal C7-10 A. albopictus cell line was infected with Wolbachia from Aa23 cells and Drosophila simulans (Riverside), resulting in cell lines designated C7-10B and C7-10R, respectively. Characterization via FISH staining showed greater stability and uniformity of Wolbachia infection in C7-10R relative to the infection in C7-10B. Characterization of the Aa23, C7-10B and C7-10R lines is discussed as a tool for the study of Wolbachia-host cell interactions.}, } @article {pmid23079772, year = {2013}, author = {Nag, JK and Shrivastava, N and Gupta, J and Misra-Bhattacharya, S}, title = {Recombinant translation initiation factor-1 of Wolbachia is an immunogenic excretory secretory protein that elicits Th2 mediated immune protection against Brugia malayi.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {36}, number = {1}, pages = {25-38}, doi = {10.1016/j.cimid.2012.09.004}, pmid = {23079772}, issn = {1878-1667}, mesh = {Animals ; B-Lymphocytes/immunology ; Brugia malayi/*immunology ; Cell Adhesion/immunology ; Cloning, Molecular ; Cross Reactions/immunology ; Cytokines/immunology/metabolism ; Cytotoxicity, Immunologic ; Elephantiasis, Filarial/*prevention & control ; Female ; Gene Expression ; Histocompatibility Antigens Class II/immunology ; Immunoglobulin E/blood/immunology ; Immunoglobulin G/blood/immunology ; Immunoglobulin M/blood/immunology ; Lymphocyte Activation ; Macrophage Activation/immunology ; Macrophages/immunology ; Male ; Mice ; Murinae ; Prokaryotic Initiation Factor-1/genetics/*immunology/metabolism ; Reactive Oxygen Species/metabolism ; Recombinant Proteins/administration & dosage/genetics/immunology ; Th2 Cells/*immunology ; Wolbachia/genetics/*immunology ; }, abstract = {Wolbachia, the intracellular alpha-proteobacteria are required for the development, fertility and survival of filarial parasites. Wolbachia Translation initiation factor-1 (Wol Tl IF-1) is one of the factors required for Wolbachia growth and viability. In the present study, we cloned, over expressed and purified Wol Tl IF-1 that exhibited strong immuno-reactivity with various categories of bancroftian sera. Immunization with the recombinant protein resulted into significant reduction in microfilarial density (70-72%) and adult worm establishment (61-63%) in susceptible Mastomys coucha. Protection offered by Wol Tl IF-1 was found associated with humoral immune arm as observed by an increased antibody level with preponderance of IgE, IgM, IgG1 and IgG2a isotypes. The anti-Wol Tl IF-1 antibodies promoted profound adherence of peritoneal exudates cells to the surface of microfilariae and infective larvae causing cytotoxicity and their death. The present study indicates potential of recombinant Wol Tl IF-1 as a promising vaccine candidate against human lymphatic filarial infection.}, } @article {pmid23077805, year = {2012}, author = {Sungpradit, S and Chatsuwan, T and Nuchprayoon, S}, title = {Susceptibility of Wolbachia, an endosymbiont of Brugia malayi microfilariae, to doxycycline determined by quantitative PCR assay.}, journal = {The Southeast Asian journal of tropical medicine and public health}, volume = {43}, number = {4}, pages = {841-850}, pmid = {23077805}, issn = {0125-1562}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/*drug effects/genetics/microbiology ; Cell Migration Assays ; Ciprofloxacin/pharmacology ; Doxycycline/*pharmacology ; Drug Resistance, Bacterial/*genetics ; Microbial Sensitivity Tests ; Microfilariae/*drug effects/genetics ; Polymerase Chain Reaction ; Rifampin/pharmacology ; Time Factors ; Wolbachia/*drug effects/genetics ; }, abstract = {Lymphatic filariasis, caused by filarial nematodes, is a mosquito-borne disease that affects over 120 million people in the tropics and subtropics. The disease is caused mainly by Wuchereria bancrofti and Brugia malayi. Fertile adult female worms release offsprings (microfilariae) into the host blood circulatory system. Transmission-blocking agents as well as antimicrobial agents have been used to reduce microfilarial density in human and animal reservoir hosts. Doxycycline and rifampicin have an effect on the obligate intracellular gram-negative bacteria, Wolbachia, which appears to exert an influence on filarial nematode embryonic and larval development, adult female fertility, and filarial survival. We investigated the effects of doxycycline, rifampicin and ciprofloxacin on B. malayi microfilarial motility, expressed as minimum effective concentration (MEC), and on Wolbachia proliferation using quantitative PCR, expressed as the concentration of the drug to inhibit bacteria growth by 50% (IC50). MEC of doxycycline was 128 and 32 microg/ml at 12 and 52 hours, respectively, but rifampicin and ciprofloxacin were ineffective (MEC >256 microg/ml). IC50 of doxycycline was 32 and 2 microg/ml at 12 and 52 hours, but this for rifampicin (8 microg/ml) and ciprofloxacin (32 microg/ml) were obtained only after 52 hour treatment. Thus, MEC and IC50 assay methods used in this study could be applied to screen other agents targeting filariae and their endosymbiont bacteria.}, } @article {pmid23071742, year = {2012}, author = {Kohnen, A and Richter, I and Brandl, R}, title = {No concordant phylogeographies of the rose gall wasp Diplolepis rosae (Hymenoptera, Cynipidae) and two associated parasitoids across Europe.}, journal = {PloS one}, volume = {7}, number = {10}, pages = {e47156}, pmid = {23071742}, issn = {1932-6203}, mesh = {Animals ; Biological Evolution ; Europe ; Haplotypes ; Phylogeny ; Phylogeography ; Population Dynamics ; Sequence Analysis, DNA ; Wasps/genetics/microbiology/*physiology ; Wolbachia/physiology ; }, abstract = {According to the Host-tracking Hypothesis, species of higher trophic levels with a close relationship to their hosts, such as parasites or parasitoids, are expected to show spatio-temporal phylogeographic patterns similar to those of their host. Alternatively, with ecological sorting, a subset of the local species pools might shift to a related host species, thereby disengaging common phylogeographic patterns. Here, we compare the phylogeographic structures of the cynipid rose gall wasp Diplolepis rosae across Europe and of two of its most common parasitoids, the wasps Orthopelma mediator and Glyphomerus stigma, by analysing the sequences of two gene fragments (COI and ITS 2). The phylogeographic structures of the three species associated with roses were incongruent. D. rosae had the lowest genetic diversity with one major clade, O. mediator showed the classical phylogeographic structure for Europe with one eastern and one western clade, and G. stigma had the highest diversity but no geographical structuring. This discordance of geographical patterns may be explained by 1) the dispersal propensity of adult parasitoids or 2) the parasitoids having the ability to switch to another host, while the primary host becomes rare or is even not available. Furthermore there was no indication that phylogenetic patterns were affected by Wolbachia infections. Our results document that communities of closely interacting species may be the result of idiosyncratic biogeographic histories.}, } @article {pmid23063745, year = {2013}, author = {Watanabe, M and Kageyama, D and Miura, K}, title = {Transfer of a parthenogenesis-inducing Wolbachia endosymbiont derived from Trichogramma dendrolimi into Trichogramma evanescens.}, journal = {Journal of invertebrate pathology}, volume = {112}, number = {1}, pages = {83-87}, doi = {10.1016/j.jip.2012.09.006}, pmid = {23063745}, issn = {1096-0805}, mesh = {Animals ; Host-Parasite Interactions/*physiology ; Parthenogenesis ; Polymerase Chain Reaction ; Wasps/*parasitology ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia, which are maternally transmitted endosymbionts, are considered to have moved horizontally between invertebrate hosts multiple times. However, it is not well understood how easily Wolbachia are transmitted horizontally between different hosts and how frequently horizontally-transmitted Wolbachia become established in their new hosts. We transferred a parthenogenesis-inducing Wolbachia endosymbiont derived from the parasitic wasp Trichogramma dendrolimi to Trichogramma evanescens. Specifically, Wolbachia was cultivated in a mosquito cell line and the Wolbachia-infected cells were microinjected into uninfected T. evanescens. Among 276 pupae inoculated with Wolbachia-infected cells, 65 adults emerged (G0). Diagnostic PCR demonstrated that 25 of 37 G0 females (68%) were Wolbachia-positive. Among isofemale lines established from G0 females, the proportions of infected lines were 80% (20 of 25) in G1 and 100% (18 of 18) in G2. In an isofemale line, infection was stably maintained for more than 10 generations. These results indicate invasion of Wolbachia into the germline of the recipient insect. Quantitative PCR demonstrated that the Wolbachia titer in the recipient host was significantly lower than that in the native host. The absence or very low number, if any, of parthenogenetically-reproducing individuals in the recipient host may be caused by the low Wolbachia titer. The Wolbachia titer in the recipients was lower in G11 than in G5, suggesting a decline in the density. Together with a previous report, our study may imply that Wolbachia in Trichogramma species are highly adapted to their hosts, which hinders robust expression of the Wolbachia phenotype in non-native host species.}, } @article {pmid23061984, year = {2012}, author = {Graham, RI and Wilson, K}, title = {Male-killing Wolbachia and mitochondrial selective sweep in a migratory African insect.}, journal = {BMC evolutionary biology}, volume = {12}, number = {}, pages = {204}, pmid = {23061984}, issn = {1471-2148}, support = {BB/F004311/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animal Migration ; Animals ; Bacterial Proteins/genetics ; Cell Nucleus/genetics ; DNA, Mitochondrial/chemistry/*genetics ; DNA, Ribosomal/chemistry/classification/genetics ; Electron Transport Complex IV/genetics ; *Evolution, Molecular ; Female ; Geography ; Host-Pathogen Interactions ; Male ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 28S/genetics ; Seasons ; Selection, Genetic ; Sequence Analysis, DNA ; Sex Factors ; Spodoptera/*genetics/microbiology ; Symbiosis ; Tanzania ; Wolbachia/classification/*genetics/physiology ; }, abstract = {BACKGROUND: Numerous recent studies have shown that resident symbiotic microorganisms of insects play a fundamental role in host ecology and evolution. The lepidopteran pest, African armyworm (Spodoptera exempta), is a highly migratory and destructive species found throughout sub-Saharan Africa, that can experience eruptive outbreaks within the space of a single generation, making predicting population dynamics and pest control forecasting extremely difficult. Three strains of Wolbachia have recently been identified infecting this species in populations sampled from Tanzania. In this study, we examined the interaction between Wolbachia pipiensis infections and the co-inherited marker, mtDNA, within populations of armyworm, as a means to investigate the population biology and evolutionary history of Wolbachia and its host.

RESULTS: A Wolbachia-infected isofemale line was established in the laboratory. Phenotypic studies confirmed the strain wExe1 as a male-killer. Partial sequencing of the mitochondrial COI gene from 164 individual field-collected armyworm of known infection status revealed 17 different haplotypes. There was a strong association between Wolbachia infection status and mtDNA haplotype, with a single dominant haplotype, haplo1 (90.2% prevalence), harbouring the endosymbiont. All three Wolbachia strains were associated with this haplotype. This indicates that Wolbachia may be driving a selective sweep on armyworm haplotype diversity. Despite very strong biological and molecular evidence that the samples represent a single species (including from nuclear 28S gene markers), the 17 haplotypes did not fall into a monophyletic clade within the Spodoptera genus; with six haplotypes (2 each from 3 geographically separate populations) differing by >11% in their nucleotide sequence to the other eleven.

CONCLUSIONS: This study suggests that three strains of Wolbachia may be driving a selective sweep on armyworm haplotype diversity, and that based on COI sequence data, S. exempta is not a monophyletic group within the Spodoptera genus. This has clear implications for the use of mtDNA as neutral genetic markers in insects, and also demonstrates the impact of Wolbachia infections on host evolutionary genetics.}, } @article {pmid23060956, year = {2012}, author = {Pan, H and Li, X and Zhang, Y}, title = {Sex affects the infection frequencies of symbionts in Bemisia tabaci.}, journal = {Communicative & integrative biology}, volume = {5}, number = {4}, pages = {337-339}, pmid = {23060956}, issn = {1942-0889}, abstract = {While biotype, host plant and geographical location are known to affect the infection dynamics of the six secondary symbionts (S-symbionts) including Hamiltonella, Arsenophonus, Cardinium, Wolbachia, Rickettsia and Fritschea in Bemisia tabaci, it remains unclear whether sex of B. tabaci has an impact on the infection frequencies of the six S-symbionts. To address this issue, gene-specific PCR were conducted to screen for the presence of the six S-symbionts in five host plant-adapted laboratory sub-populations with the same genetic background. Significant variations were exhibited in the infection rates of Rickettsia, Cardinium, Rickettsia + Hamiltonella (RH), Rickettsia + Cardinium (RC), Hamiltonella + Cardinium (HC) and Rickettsia + Hamiltonella + Cardinium (RHC) among the five host plant-adapted sub-populations. Moreover, Rickettsia, Hamiltonella, Cardinium, RH, RC, HC and RHC were present at a significantly higher frequency in the females than in the males of the five host plant-adapted sub-populations. This indicates that sex is another important factor affecting the population dynamics of S-symbionts in B. tabaci.}, } @article {pmid23056182, year = {2012}, author = {Dematheis, F and Kurtz, B and Vidal, S and Smalla, K}, title = {Microbial communities associated with the larval gut and eggs of the Western corn rootworm.}, journal = {PloS one}, volume = {7}, number = {10}, pages = {e44685}, pmid = {23056182}, issn = {1932-6203}, mesh = {Animals ; Bacteria/classification/genetics/growth & development ; Cluster Analysis ; Coleoptera/genetics/*microbiology/physiology ; DNA, Bacterial/analysis/genetics ; DNA, Fungal/analysis/genetics ; Female ; Fungi/classification/genetics/physiology ; Gastrointestinal Tract/*microbiology ; Host-Parasite Interactions ; Host-Pathogen Interactions ; Larva/genetics/microbiology/physiology ; Metagenome/*genetics ; Metagenomics/methods ; Ovum/*microbiology ; Phylogeny ; Plant Roots/parasitology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rhizosphere ; Soil Microbiology ; Zea mays/*parasitology ; }, abstract = {BACKGROUND: The western corn rootworm (WCR) is one of the economically most important pests of maize. A better understanding of microbial communities associated with guts and eggs of the WCR is required in order to develop new pest control strategies, and to assess the potential role of the WCR in the dissemination of microorganisms, e.g., mycotoxin-producing fungi.

Total community (TC) DNA was extracted from maize rhizosphere, WCR eggs, and guts of larvae feeding on maize roots grown in three different soil types. Denaturing gradient gel electrophoresis (DGGE) and sequencing of 16S rRNA gene and ITS fragments, PCR-amplified from TC DNA, were used to investigate the fungal and bacterial communities, respectively. Microorganisms in the WCR gut were not influenced by the soil type. Dominant fungal populations in the gut were affiliated to Fusarium spp., while Wolbachia was the most abundant bacterial genus. Identical ribosomal sequences from gut and egg samples confirmed a transovarial transmission of Wolbachia sp. Betaproteobacterial DGGE indicated a stable association of Herbaspirillum sp. with the WCR gut. Dominant egg-associated microorganisms were the bacterium Wolbachia sp. and the fungus Mortierella gamsii.

CONCLUSION/SIGNIFICANCE: The soil type-independent composition of the microbial communities in the WCR gut and the dominance of only a few microbial populations suggested either a highly selective environment in the gut lumen or a high abundance of intracellular microorganisms in the gut epithelium. The dominance of Fusarium species in the guts indicated WCR larvae as vectors of mycotoxin-producing fungi. The stable association of Herbaspirillum sp. with WCR gut systems and the absence of corresponding sequences in WCR eggs suggested that this bacterium was postnatally acquired from the environment. The present study provided new insights into the microbial communities associated with larval guts and eggs of the WCR. However, their biological role remains to be explored.}, } @article {pmid23055615, year = {2012}, author = {Uday, J and Puttaraju, HP}, title = {Comparative analysis of Wolbachia surface protein in D. melanoagster, A. tabida and B. malayi.}, journal = {Bioinformation}, volume = {8}, number = {15}, pages = {711-715}, pmid = {23055615}, issn = {0973-2063}, abstract = {Wolbachia surface protein (WSP) is an eight beta-barrel transmembrane structure which participates in host immune response, cell proliferation, pathogenicity and controlled cell death program. The protein has four extracellular loops containing hyper variable regions separated by conserved regions. The WSP structure is homologous to Neisseria surface protein (Nsp A) which has about 34% similarity including antigenic variation and hydrophilicity. Recombination has a large impact on diversity of this protein including positive selection which is major constraint on protein evolution. The molecular mechanism through which Wolbachia induces various reproductive anomalies is unclear; a key feature observed for such anomalies might be because of Wolbachia undergoing extensive recombination. In Wolbachia, increased recombination is observed in ankyrin proteins, surface proteins and in some hypothetical proteins. Genetic divergence is extensive in the WSP gene, WSP is known to be a chimeric protein involved in host-symbiont interactions. Here we predicted the structural and functional variations in WSP sequences of Wolbachia present in D. melanogaster, A. tabida and in B. malayi.}, } @article {pmid23053393, year = {2012}, author = {Babu, S and Nutman, TB}, title = {Immunopathogenesis of lymphatic filarial disease.}, journal = {Seminars in immunopathology}, volume = {34}, number = {6}, pages = {847-861}, pmid = {23053393}, issn = {1863-2300}, support = {Y99 AI999999/AI/NIAID NIH HHS/United States ; Z99 AI999999/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Adaptive Immunity ; Animals ; Elephantiasis, Filarial/complications/diagnosis/*immunology ; Humans ; Immunity, Innate ; Lymphedema/etiology ; }, abstract = {Although two thirds of the 120 million people infected with lymph-dwelling filarial parasites have subclinical infections, ~40 million have lymphedema and/or other pathologic manifestations including hydroceles (and other forms of urogenital disease), episodic adenolymphangitis, tropical pulmonary eosinophilia, lymphedema, and (in its most severe form) elephantiasis. Adult filarial worms reside in the lymphatics and lymph nodes and induce changes that result in dilatation of lymphatics and thickening of the lymphatic vessel walls. Progressive lymphatic damage and pathology results from the summation of the effect of tissue alterations induced by both living and nonliving adult parasites, the host inflammatory response to the parasites and their secreted antigens, the host inflammatory response to the endosymbiont Wolbachia, and those seen as a consequence of secondary bacterial or fungal infections. Inflammatory damage induced by filarial parasites appears to be multifactorial, with endogenous parasite products, Wolbachia, and host immunity all playing important roles. This review will initially examine the prototypical immune responses engendered by the parasite and delineate the regulatory mechanisms elicited to prevent immune-mediated pathology. This will be followed by a discussion of the proposed mechanisms underlying pathogenesis, with the central theme being that pathogenesis is a two-step process-the first initiated by the parasite and host innate immune system and the second propagated mainly by the host's adaptive immune system and by other factors (including secondary infections).}, } @article {pmid23049857, year = {2012}, author = {McNulty, SN and Abubucker, S and Simon, GM and Mitreva, M and McNulty, NP and Fischer, K and Curtis, KC and Brattig, NW and Weil, GJ and Fischer, PU}, title = {Transcriptomic and proteomic analyses of a Wolbachia-free filarial parasite provide evidence of trans-kingdom horizontal gene transfer.}, journal = {PloS one}, volume = {7}, number = {9}, pages = {e45777}, pmid = {23049857}, issn = {1932-6203}, support = {R01 AI081803/AI/NIAID NIH HHS/United States ; T32 AI007172/AI/NIAID NIH HHS/United States ; T32-AI007172/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi ; Conserved Sequence ; Gene Expression Regulation ; *Gene Transfer, Horizontal ; Genome ; Gerbillinae ; Immunohistochemistry/methods ; Mass Spectrometry/methods ; Onchocerca/*microbiology ; Parasites/genetics ; Protein Structure, Tertiary ; Proteomics/*methods ; Symbiosis/genetics ; *Transcription, Genetic ; Transcriptome ; Wolbachia/*genetics ; }, abstract = {Most filarial parasites in the subfamilies Onchocercinae and Dirofilariinae depend on Wolbachia endobacteria to successfully carry out their life cycle. Recently published data indicate that the few Wolbachia-free species in these subfamilies were infected in the distant past and have subsequently shed their endosymbionts. We used an integrated transcriptomic and proteomic analysis of Onchocerca flexuosa to explore the molecular mechanisms that allow worms of this species to survive without a bacterial partner. Roche/454 sequencing of the adult transcriptome produced 16,814 isogroup and 47,252 singleton sequences that are estimated to represent approximately 41% of the complete gene set. Sequences similar to 97 Wolbachia genes were identified from the transcriptome, some of which appear on the same transcripts as sequences similar to nematode genes. Computationally predicted peptides, including those with similarity to Wolbachia proteins, were classified at the domain and pathway levels in order to assess the metabolic capabilities of O. flexuosa and compare against the Wolbachia-dependent model filaria, Brugia malayi. Transcript data further facilitated a shotgun proteomic analysis of O. flexuosa adult worm lysate, resulting in the identification of 1,803 proteins. Three of the peptides detected by mass spectroscopy map to two ABC transport-related proteins from Wolbachia. Antibodies raised to one of the Wolbachia-like peptides labeled a single 38 kDa band on Western blots of O. flexuosa lysate and stained specific worm tissues by immunohistology. Future studies will be required to determine the exact functions of Wolbachia-like peptides and proteins in O. flexuosa and to assess their roles in worm biology.}, } @article {pmid23042170, year = {2012}, author = {Morse, SF and Dick, CW and Patterson, BD and Dittmar, K}, title = {Some like it hot: evolution and ecology of novel endosymbionts in bat flies of cave-roosting bats (hippoboscoidea, nycterophiliinae).}, journal = {Applied and environmental microbiology}, volume = {78}, number = {24}, pages = {8639-8649}, pmid = {23042170}, issn = {1098-5336}, mesh = {Animals ; Bacterial Physiological Phenomena ; Chiroptera/*parasitology ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Diptera/*microbiology ; Gammaproteobacteria/*classification/genetics/*isolation & purification ; Heat-Shock Proteins/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/classification/genetics/isolation & purification ; }, abstract = {We investigated previously unknown associations between bacterial endosymbionts and bat flies of the subfamily Nycterophiliinae (Diptera, Streblidae). Molecular analyses revealed a novel clade of Gammaproteobacteria in Nycterophilia bat flies. This clade was not closely related to Arsenophonus-like microbes found in its sister genus Phalconomus and other bat flies. High population infection rates in Nycterophilia across a wide geographic area, the presence of the symbionts in pupae, the general codivergence between hosts and symbionts, and high AT composition bias in symbiont genes together suggest that this host-symbiont association is obligate in nature and ancient in origin. Some Nycterophilia samples (14.8%) also contained Wolbachia supergroup F (Alphaproteobacteria), suggesting a facultative symbiosis. Likelihood-based ancestral character mapping revealed that, initially, obligate symbionts exhibited association with host-specific Nycterophilia bat flies that use a broad temperature range of cave environments for pupal development. As this mutualism evolved, the temperature range of bat flies narrowed to an exclusive use of hot caves, which was followed by a secondary broadening of the bat flies' host associations. These results suggest that the symbiosis has influenced the environmental tolerance of parasite life history stages. Furthermore, the contingent change to an expanded host range of Nycterophilia bat flies upon narrowing the ecological niche of their developmental stages suggests that altered environmental tolerance across life history stages may be a crucial factor in shaping parasite-host relationships.}, } @article {pmid23041355, year = {2012}, author = {Lefoulon, E and Gavotte, L and Junker, K and Barbuto, M and Uni, S and Landmann, F and Laaksonen, S and Saari, S and Nikander, S and de Souza Lima, S and Casiraghi, M and Bain, O and Martin, C}, title = {A new type F Wolbachia from Splendidofilariinae (Onchocercidae) supports the recent emergence of this supergroup.}, journal = {International journal for parasitology}, volume = {42}, number = {11}, pages = {1025-1036}, doi = {10.1016/j.ijpara.2012.09.004}, pmid = {23041355}, issn = {1879-0135}, mesh = {Animals ; DNA, Bacterial/genetics ; Gene Expression Regulation, Bacterial ; Genetic Variation ; Molecular Sequence Data ; Nematoda/*microbiology ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal/genetics/metabolism ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia are vertically transmitted endosymbiotic bacteria of arthropods and onchocercid nematodes. It is commonly accepted that they co-evolved with their filarial hosts, and have secondarily been lost in some species. However, most of the data on the Wolbachia/Onchocercidae relationship have been derived from studies on two subfamilies, the Dirofilariinae and the Onchocercinae, which harbour parasites of humans and domestic animals. Within the last few years, analyses of more diverse material have suggested that some groups of Onchocercidae do not have Wolbachia, such as recently studied Splendidofilariinae from birds. This study takes advantage of the analysis of additional Splendidofilariinae, Rumenfilaria andersoni from a Finnish reindeer and Madathamugadia hiepei from a South African gecko, using PCR, immunohistochemical staining and whole-mount fluorescent analysis to detect Wolbachia and describe its strains. A DNA barcoding approach and phylogenetic analyses were used to investigate the symbiosis between Wolbachia and the Onchocercidae. A new supergroup F Wolbachia was demonstrated in M. hiepei, representing the first filarial nematode harbouring Wolbachia described in a non-mammalian host. In the adult, Wolbachia infects the female germline but not the hypodermis, and intestinal cells are also infected. The phylogenetic analyses confirmed a recent emergence of supergroup F. They also suggested several events of horizontal transmission between nematodes and arthropods in this supergroup, and the existence of different metabolic interactions between the filarial nematodes and their symbionts.}, } @article {pmid23036316, year = {2013}, author = {Laurin-Lemay, S and Angers, B and Benrey, B and Brodeur, J}, title = {Inconsistent genetic structure among members of a multitrophic system: did bruchid parasitoids (Horismenus spp.) escape the effects of bean domestication?.}, journal = {Bulletin of entomological research}, volume = {103}, number = {2}, pages = {182-192}, doi = {10.1017/S000748531200051X}, pmid = {23036316}, issn = {1475-2670}, mesh = {Animals ; *Biological Evolution ; Coleoptera/*genetics/parasitology ; *Food Chain ; Genetic Variation ; Host-Parasite Interactions ; Mexico ; Phaseolus/*genetics ; Phylogeography ; Wasps/*genetics/microbiology ; Wolbachia/isolation & purification ; }, abstract = {Anthropogenic range expansion and cultural practices have modified the distribution, abundance and genetic diversity of domesticated organisms, thereby altering multitrophic assemblages through space and time. The putative Mesoamerican domestication centre of the common bean, Phaseolus vulgaris L., in Mexico allows investigating the effects of plant domestication on the genetic structure of members of a multitrophic system. The aim of this study was to compare the evolutionary history of Horismenus parasitoids (Hymenoptera: Eulophidae) to those of their bruchid beetle hosts (Coleoptera: Bruchidae) and their domesticated host plant (P. vulgaris), in the context of traditional agriculture in Mexico. We analyzed the population genetic structure of four Horismenus species in Mexico using mitochondrial COI haplotype data. The two most abundant parasitoid species were Horismenus depressus and Horismenus missouriensis. Horismenus missouriensis were infected by Wolbachia endosymbionts and had little to no population differentiation (F(ST) = 0.06). We suspect the mitochondrial history of H. missouriensis to be blurred by Wolbachia, because differentiation among infected vs. non-infected individuals exists (F(ST) = 0.11). Populations of H. depressus were found to be highly differentiated (F(ST) = 0.34), but the genetic structuring could not be explained by tested spatial components. We then compared the genetic structure observed in this parasitoid species to previously published studies on bruchid beetles and their host plants. Despite extensive human-mediated migration and likely population homogenization of its two Acanthoscelides bruchid beetle hosts, H. depressus populations are structured like its host plant, by a recent dispersal from a diverse ancestral gene pool. Distinct evolutionary dynamics may explain inconsistent patterns among trophic levels. Parasitoids likely migrate from wild bean populations and are poorly adapted to bean storage conditions similar to their bruchid beetle hosts. Integrating several trophic levels to the study of evolutionary history has proven to be fruitful in detecting different ecological responses to human-mediated disturbances and host parasite interactions.}, } @article {pmid23028533, year = {2012}, author = {Palavesam, A and Guerrero, FD and Heekin, AM and Wang, J and Dowd, SE and Sun, Y and Foil, LD and Pérez de León, AA}, title = {Pyrosequencing-based analysis of the microbiome associated with the horn fly, Haematobia irritans.}, journal = {PloS one}, volume = {7}, number = {9}, pages = {e44390}, pmid = {23028533}, issn = {1932-6203}, mesh = {Animals ; Bacteria/*genetics ; Cattle ; DNA, Ribosomal/genetics ; Female ; Male ; Metagenome/*genetics ; Muscidae/*microbiology ; *Sequence Analysis, DNA ; Wolbachia/genetics ; }, abstract = {The horn fly, Haematobia irritans, is one of the most economically important pests of cattle. Insecticides have been a major element of horn fly management programs. Growing concerns with insecticide resistance, insecticide residues on farm products, and non-availability of new generation insecticides, are serious issues for the livestock industry. Alternative horn fly control methods offer the promise to decrease the use of insecticides and reduce the amount of insecticide residues on livestock products and give an impetus to the organic livestock farming segment. The horn fly, an obligatory blood feeder, requires the help of microflora to supply additional nutrients and metabolize the blood meal. Recent advancements in DNA sequencing methodologies enable researchers to examine the microflora diversity independent of culture methods. We used the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) method to carry out the classification analysis of bacterial flora in adult female and male horn flies and horn fly eggs. The bTEFAP method identified 16S rDNA sequences in our samples which allowed the identification of various prokaryotic taxa associated with the life stage examined. This is the first comprehensive report of bacterial flora associated with the horn fly using a culture-independent method. Several rumen, environmental, symbiotic and pathogenic bacteria associated with the horn fly were identified and quantified. This is the first report of the presence of Wolbachia in horn flies of USA origin and is the first report of the presence of Rikenella in an obligatory blood feeding insect.}, } @article {pmid23028321, year = {2012}, author = {Serbus, LR and Landmann, F and Bray, WM and White, PM and Ruybal, J and Lokey, RS and Debec, A and Sullivan, W}, title = {A cell-based screen reveals that the albendazole metabolite, albendazole sulfone, targets Wolbachia.}, journal = {PLoS pathogens}, volume = {8}, number = {9}, pages = {e1002922}, pmid = {23028321}, issn = {1553-7374}, support = {F32 GM080192/GM/NIGMS NIH HHS/United States ; R25 GM058903/GM/NIGMS NIH HHS/United States ; S10 RR022455/RR/NCRR NIH HHS/United States ; }, mesh = {Albendazole/*analogs & derivatives/pharmacology ; Animals ; Brugia malayi/drug effects/*microbiology ; Cell Line ; Drosophila melanogaster/drug effects/*microbiology ; Filariasis/*drug therapy ; Microbial Sensitivity Tests ; Microtubules/drug effects ; Symbiosis ; Wolbachia/*drug effects ; }, abstract = {Wolbachia endosymbionts carried by filarial nematodes give rise to the neglected diseases African river blindness and lymphatic filariasis afflicting millions worldwide. Here we identify new Wolbachia-disrupting compounds by conducting high-throughput cell-based chemical screens using a Wolbachia-infected, fluorescently labeled Drosophila cell line. This screen yielded several Wolbachia-disrupting compounds including three that resembled Albendazole, a widely used anthelmintic drug that targets nematode microtubules. Follow-up studies demonstrate that a common Albendazole metabolite, Albendazole sulfone, reduces intracellular Wolbachia titer both in Drosophila melanogaster and Brugia malayi, the nematode responsible for lymphatic filariasis. Significantly, Albendazole sulfone does not disrupt Drosophila microtubule organization, suggesting that this compound reduces titer through direct targeting of Wolbachia. Accordingly, both DNA staining and FtsZ immunofluorescence demonstrates that Albendazole sulfone treatment induces Wolbachia elongation, a phenotype indicative of binary fission defects. This suggests that the efficacy of Albendazole in treating filarial nematode-based diseases is attributable to dual targeting of nematode microtubules and their Wolbachia endosymbionts.}, } @article {pmid23025192, year = {2012}, author = {Beckmann, JF and Fallon, AM}, title = {Decapitation improves detection of Wolbachia pipientis (Rickettsiales: Anaplasmataceae) in Culex pipiens (Diptera: Culicidae) mosquitoes by the polymerase chain reaction.}, journal = {Journal of medical entomology}, volume = {49}, number = {5}, pages = {1103-1108}, pmid = {23025192}, issn = {0022-2585}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cricetinae ; Culex/*microbiology ; Decapitation ; False Negative Reactions ; Polymerase Chain Reaction/standards ; Wolbachia/*isolation & purification ; }, abstract = {Polymerase chain reaction (PCR) is often used to detect microorganisms, pathogens, or both, including the reproductive parasite Wolbachia pipientis (Rickettsiales: Anaplasmataceae), in mosquitoes. Natural populations of Culex pipiens L. (Diptera: Culicidae) mosquitoes are infected with one or more strains of W. pipientis, and crosses between mosquitoes harboring different Wolbachia strains provide one of the best-known examples of cytoplasmic incompatibililty (CI). When we used PCR to monitor Wolbachia in the Buckeye strain of Culex pipiens, and in a Wolbachia-cured sister colony obtained by tetracycline treatment, we noted false negative PCR reactions with DNA samples from infected mosquitoes; these results were inconsistent with direct microscopic observation of Wolbachia-like particles in gonads dissected from mosquitoes in the same population. Assays with diluted template often improved detection of positive samples, suggesting that DNA prepared from whole mosquitoes contained an inhibitor of the PCR reaction. We reconciled discrepancies between PCR and microscopy by systematic measurement of the PCR reaction in the presence of an internal standard. Mosquito decapitation before DNA extraction restored the reliability of the PCR reaction, allowing accurate determination of Wolbachia infection status in infected and tetracycline-cured mosquito populations, consistent with microscopic examination. Using PCR primers based on the Tr1 gene, we confirmed that the Wolbachia infection in the Buckeye strain of Culex pipiens belongs to the genotype designated wPip1. Finally, to explore more widely the distribution of PCR inhibitors, we demonstrated that DNA isolated from the cricket, Acheta domesticus (L.); the beetle, Tenebrio molitor L.; the honey bee, Apis mellifera L.; and the mosquito, Anopheles punctipennis Say also contained PCR inhibitors. These results underscore the importance of measuring the presence of inhibitors in PCR templates by using a known positive standard, and provide an approach that will facilitate use of PCR to monitor environmental samples of mosquitoes that harbor endosymbionts or pathogenic organisms.}, } @article {pmid23016971, year = {2012}, author = {Gilbert, SF}, title = {Ecological developmental biology: environmental signals for normal animal development.}, journal = {Evolution & development}, volume = {14}, number = {1}, pages = {20-28}, doi = {10.1111/j.1525-142X.2011.00519.x}, pmid = {23016971}, issn = {1525-142X}, mesh = {Animals ; *Biological Evolution ; Developmental Biology ; Ecology ; *Embryonic Development ; *Environment ; *Gene-Environment Interaction ; Symbiosis ; }, abstract = {The environment plays instructive roles in development and selective roles in evolution. This essay reviews several of the instructive roles whereby the organism has evolved to receive cues from the environment in order to modulate its developmental trajectory. The environmental cues can be abiotic (such as temperature or photoperiod) or biotic (such as those emanating from predators, conspecifics, or food), and the "alteration" produces a normal, not a pathological, phenotype, that is appropriate for the environment. In addition, symbiotic organisms can produce important signals during normal development. Environmental cues can be obligatory, such that the organism cannot develop without the environmental cue. These cues often permit and instruct the organism to proceed from one developmental stage to another, as when larvae receive cues to settle and undergo metamorphosis from substrates. Such obligatory cues can also be given by symbionts, as when Wolbachia bacteria prevent apoptosis in developing ovaries of some wasps. Other environmental cues can be used facultatively, allowing organisms to follow different developmental trajectories depending on whether the cue is present or not. This can be seen in the temperature-dependent determination of sex in many reptiles and in the determination of thermotolerance in aphids by their symbiotic bacteria. Signaling from the environment is essential in development, and co-development appears to be normative between symbionts and their hosts. Here, one sees the reciprocal induction of gene expression, just as within the embryonic organism. The ability of organisms to respond to environmental cues by producing different phenotypes may be critically important in evolution, and it may be an essential feature that can facilitate or limit evolution.}, } @article {pmid22990977, year = {2012}, author = {Maciel-de-Freitas, R and Aguiar, R and Bruno, RV and Guimarães, MC and Lourenço-de-Oliveira, R and Sorgine, MH and Struchiner, CJ and Valle, D and O'Neill, SL and Moreira, LA}, title = {Why do we need alternative tools to control mosquito-borne diseases in Latin America?.}, journal = {Memorias do Instituto Oswaldo Cruz}, volume = {107}, number = {6}, pages = {828-829}, doi = {10.1590/s0074-02762012000600021}, pmid = {22990977}, issn = {1678-8060}, mesh = {Alphavirus Infections/prevention & control ; Animals ; Chikungunya Fever ; Culicidae/*microbiology ; Dengue/prevention & control ; Humans ; Insect Vectors/*microbiology ; Latin America ; Malaria/prevention & control ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, abstract = {In this opinion paper, we discuss the potential and challenges of using the symbiont Wolbachia to block mosquito transmitted diseases such as dengue, malaria and chikungunya in Latin America.}, } @article {pmid22984624, year = {2012}, author = {Liu, S and Chougule, NP and Vijayendran, D and Bonning, BC}, title = {Deep sequencing of the transcriptomes of soybean aphid and associated endosymbionts.}, journal = {PloS one}, volume = {7}, number = {9}, pages = {e45161}, pmid = {22984624}, issn = {1932-6203}, mesh = {Amino Acid Sequence ; Animals ; Aphids/*genetics/microbiology/physiology ; Base Sequence ; Buchnera/genetics/physiology ; Digestive System/metabolism/microbiology ; Enterobacteriaceae/genetics/physiology ; Genes, Bacterial/genetics ; Genes, Insect/genetics ; High-Throughput Nucleotide Sequencing/*methods ; Host-Pathogen Interactions ; Insect Proteins/classification/genetics ; Molecular Sequence Data ; Pisum sativum/parasitology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid ; Glycine max/parasitology ; *Transcriptome ; Wolbachia/genetics/physiology ; }, abstract = {BACKGROUND: The soybean aphid has significantly impacted soybean production in the U.S. Transcriptomic analyses were conducted for further insight into leads for potential novel management strategies.

Transcriptomic data were generated from whole aphids and from 2,000 aphid guts using an Illumina GAII sequencer. The sequence data were assembled de novo using the Velvet assembler. In addition to providing a general overview, we demonstrate (i) the use of the Multiple-k/Multiple-C method for de novo assembly of short read sequences, followed by BLAST annotation of contigs for increased transcript identification: From 400,000 contigs analyzed, 16,257 non-redundant BLAST hits were identified; (ii) analysis of species distributions of top non-redundant hits: 80% of BLAST hits (minimum e-value of 1.0-E3) were to the pea aphid or other aphid species, representing about half of the pea aphid genes; (iii) comparison of relative depth of sequence coverage to relative transcript abundance for genes with high (membrane alanyl aminopeptidase N) or low transcript abundance; (iv) analysis of the Buchnera transcriptome: Transcripts from 57.6% of the genes from Buchnera aphidicola were identified; (v) identification of Arsenophonus and Wolbachia as potential secondary endosymbionts; (vi) alignment of full length sequences from RNA-seq data for the putative salivary gland protein C002, the silencing of which has potential for aphid management, and the putative Bacillus thuringiensis Cry toxin receptors, aminopeptidase N and alkaline phosphatase.

CONCLUSIONS/SIGNIFICANCE: THIS STUDY PROVIDES THE MOST COMPREHENSIVE DATA SET TO DATE FOR SOYBEAN APHID GENE EXPRESSION: This work also illustrates the utility of short-read transcriptome sequencing and the Multiple-k/Multiple-C method followed by BLAST annotation for rapid identification of target genes for organisms for which reference genome sequences are not available, and extends the utility to include the transcriptomes of endosymbionts.}, } @article {pmid22973543, year = {2012}, author = {Aw, WC and Dowell, FE and Ballard, JW}, title = {Using near-infrared spectroscopy to resolve the species, gender, age, and the presence of Wolbachia infection in laboratory-reared Drosophila.}, journal = {G3 (Bethesda, Md.)}, volume = {2}, number = {9}, pages = {1057-1065}, pmid = {22973543}, issn = {2160-1836}, mesh = {Animals ; Drosophila/*classification/*microbiology ; Female ; Male ; *Sex Characteristics ; *Spectroscopy, Near-Infrared ; *Wolbachia ; }, abstract = {The aim of the study was to determine the accuracy of near-infrared spectroscopy (NIRS) in determining species, gender, age, and the presence of the common endosymbiont Wolbachia in laboratory-reared Drosophila. NIRS measures the absorption of light by organic molecules. Initially, a calibration model was developed for each study. An independent set with flies not involved in initial cross-validation was then used to validate the accuracy of each calibration model. Flies from the independent sets were correctly classified into Drosophila melanogaster and Drosophila simulans with 94% and 82% accuracy, respectively, whereas flies were successfully classified by gender with accuracy greater than 90%. In the age grading test, correlation plots of the actual and predicted age for males and females of D. melanogaster and D. simulans were shown to be overlapping between the adjacent age groups. It is, however, possible to predict the age of flies as less than 9 days of age with 62-88% accuracy and flies that are equal to or older than 9 days of age with 91-98% accuracy. Finally, we used NIRS to detect the presence of Wolbachia in flies. Flies from the independent sets were successfully identified as infected or not infected with Wolbachia with approximately 90% accuracy. These results suggest that NIRS has the potential to quantify the species, gender, and presence of Wolbachia in fly populations. However, additional optimization of the protocol may be necessary before the technique can reliably estimate fly age.}, } @article {pmid22961037, year = {2012}, author = {Martínez, H and Toledo, J and Liedo, P and Mateos, M}, title = {Survey of heritable endosymbionts in southern Mexico populations of the fruit fly species Anastrepha striata and A. ludens.}, journal = {Current microbiology}, volume = {65}, number = {6}, pages = {711-718}, pmid = {22961037}, issn = {1432-0991}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Mexico ; Multilocus Sequence Typing ; Phylogeny ; Symbiosis/*genetics ; Tephritidae/classification/genetics/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Heritable endosymbiotic bacteria associated with insects are ubiquitous and taxonomically diverse. Many of these endosymbionts influence the fitness of their hosts and/or manipulate their host reproduction. Exploiting the effects of endosymbionts on hosts for pest control is a growing research area, but requires knowledge of endosymbionts associated with the target pest population. In this study, we used molecular methods to screen southern Mexico populations of two species of tephritid fruit fly pests, Anastrepha ludens and A. striata, for heritable bacteria. The only heritable endosymbiont found was Wolbachia in A. striata. Based on multilocus sequence typing and phylogenetic analyses, this Wolbachia strain is new and belongs to the Wolbachia supergroup B. Wolbachia strains previously reported in members of the genus Anastrepha in South America belong to supergroup A. We discuss the potential implications for pest control of the presence of a different Wolbachia strain in southern Mexico.}, } @article {pmid22953011, year = {2012}, author = {McNaughton, D}, title = {The importance of long-term social research in enabling participation and developing engagement strategies for new dengue control technologies.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {8}, pages = {e1785}, pmid = {22953011}, issn = {1935-2735}, mesh = {Adolescent ; Aedes/microbiology ; Aged ; Aged, 80 and over ; Animals ; Australia ; Child ; Child, Preschool ; Dengue/*prevention & control ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Mosquito Control/*methods ; Patient Acceptance of Health Care/*psychology ; Wolbachia/growth & development ; Young Adult ; }, abstract = {BACKGROUND: In recent years, new strategies aimed at reducing the capacity of mosquito vectors to transmit dengue fever have emerged. As with earlier control methods, they will have to be employed in a diverse range of communities across the globe and into the main settings for disease transmission, the homes, businesses and public buildings of residents in dengue-affected areas. However, these strategies are notably different from previous methods and draw on technologies that are not without controversy. Public engagement and authorization are critical to the future success of these programs.

This paper reports on an Australian case study where long-term social research was used to enable participation and the design of an engagement strategy tailored specifically to the sociopolitical setting of a potential trial release site of Wolbachia-infected Aedes aegytpi mosquitoes. Central themes of the social research, methods used and conclusions drawn are briefly described. Results indicate that different communities are likely to have divergent expectations, concerns and cultural sensibilities with regard to participation, engagement and authorization.

CONCLUSIONS/SIGNIFICANCE: The findings show that a range of issues need to be understood and taken into account to enable sensitive, ethical and effective engagement when seeking public support for new dengue control methods.}, } @article {pmid22951738, year = {2012}, author = {Duron, O and Bernard, J and Atyame, CM and Dumas, E and Weill, M}, title = {Rapid evolution of Wolbachia incompatibility types.}, journal = {Proceedings. Biological sciences}, volume = {279}, number = {1746}, pages = {4473-4480}, pmid = {22951738}, issn = {1471-2954}, mesh = {Animals ; Bacterial Proteins/*genetics ; Culex/*microbiology/physiology ; Cytoplasm/microbiology ; *Evolution, Molecular ; Female ; Fertility ; Host-Pathogen Interactions ; Hybridization, Genetic ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Reproduction ; Sequence Alignment ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {In most insects, the endosymbiont Wolbachia induces cytoplasmic incompatibility (CI), an embryonic mortality observed when infected males mate either with uninfected females or with females infected by an incompatible Wolbachia strain. Although the molecular mechanism of CI remains elusive, it is classically viewed as a modification-rescue model, in which a Wolbachia mod function disables the reproductive success of the sperm of infected males, unless eggs are infected and express a compatible resc function. The extent to which the modification-rescue model can predict highly complex CI pattern remains a challenging issue. Here, we show the rapid evolution of the mod-resc system in the Culex pipiens mosquito. We have surveyed four incompatible laboratory isofemale lines over 50 generations and observed in two of them that CI has evolved from complete to partial incompatibility (i.e. the production of a mixture of compatible and incompatible clutches). Emergence of the new CI types depends only on Wolbachia determinants and can be simply explained by the gain of new resc functions. Evolution of CI types in Cx. pipiens thus appears as a gradual process, in which one or several resc functions can coexist in the same individual host in addition to the ones involved in the self-compatibility. Our data identified CI as a very dynamic process. We suggest that ancestral and mutant Wolbachia expressing distinct resc functions can co-infect individual hosts, opening the possibility for the mod functions to evolve subsequently. This gives a first clue towards the understanding of how Wolbachia reached highly complex CI pattern in host populations.}, } @article {pmid22947080, year = {2012}, author = {Zug, R and Koehncke, A and Hammerstein, P}, title = {Epidemiology in evolutionary time: the case of Wolbachia horizontal transmission between arthropod host species.}, journal = {Journal of evolutionary biology}, volume = {25}, number = {11}, pages = {2149-2160}, doi = {10.1111/j.1420-9101.2012.02601.x}, pmid = {22947080}, issn = {1420-9101}, mesh = {Algorithms ; Animals ; Arthropods/*microbiology/physiology ; *Biological Evolution ; Computer Simulation ; Models, Biological ; Phylogeny ; Species Specificity ; Stochastic Processes ; Time Factors ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachia are bacterial endosymbionts that manipulate the reproduction of their arthropod hosts. Although theory suggests that infections are frequently lost within host species due to the evolution of resistance, Wolbachia infect a huge number of species worldwide. This apparent paradox suggests that horizontal transmission between host species has been a key factor in shaping the global Wolbachia pandemic. Because Wolbachia infections are thus acquired and lost like any other infection, we use a standard epidemiological model to analyse Wolbachia horizontal transmission dynamics over evolutionary time. Conceptually modifying the model, we apply it not to transmission between individuals but between species. Because, on evolutionary timescales, infections spread frequently between closely related species and occasionally over large phylogenetic distances, we represent the set of host species as a small-world network that satisfies both requirements. Our model reproduces the effect of basic epidemiological parameters, which demonstrates the validity of our approach. We find that the ratio between transmission rate and recovery rate is crucial for determining the proportion of infected species (incidence) and that, in a given host network, the incidence may still be increasing over evolutionary time. Our results also point to the importance of occasional transmission over long phylogenetic distances for the observed high incidence levels of Wolbachia. In conclusion, we are able to explain why Wolbachia are so abundant among arthropods, although selection for resistance within hosts often leads to infection loss. Furthermore, our unorthodox approach of using epidemiology in evolutionary time can be applied to all symbionts that use horizontal transmission to infect new hosts.}, } @article {pmid22945464, year = {2012}, author = {Nachappa, P and Levy, J and Tamborindeguy, C}, title = {Transcriptome analyses of Bactericera cockerelli adults in response to "Candidatus Liberibacter solanacearum" infection.}, journal = {Molecular genetics and genomics : MGG}, volume = {287}, number = {10}, pages = {803-817}, pmid = {22945464}, issn = {1617-4623}, mesh = {Aging ; Animals ; Gene Expression Profiling ; *Helicobacter ; Hemiptera/*genetics/microbiology ; Insect Proteins/analysis/*genetics ; }, abstract = {The potato/tomato psyllid, Bactericera cockerelli (Šulc) is an economically important crop pest that not only causes damage through its feeding but also transmits the bacterium, "Candidatus Liberibacter solanacearum" (CLs), which causes zebra chip disease in potato. There is some information about the phenotypic effects of phytopathogenic bacteria on their insect vectors; however, there are no published reports of the molecular mechanisms underlying phytopathogenic bacteria-insect vector interaction. In order to investigate the effects of CLs infection on B. cockerelli, transcriptomic analyses of CLs-infected and uninfected adult psyllids that were reared on potato were performed. De novo assembly of cDNA sequences generated 136,518 and 109,983 contigs for infected and uninfected insect libraries with an average contig length of 514 bp. BlastX analysis against the NCBI-nr database revealed that 33.33 % had significant matches. Gene ontology data illustrated that the majority of the expressed psyllid genes are involved in metabolic process, biological regulation, binding and catalytic activity. The psyllid transcriptome had an abundance of genes such as vitellogenin, heat shock protein, ejaculatory bulb-specific protein, ferritin, and cytochrome oxidase. Notably absent in the psyllid transcriptome were innate immunity genes induced in response to Gram-negative bacteria (IMD pathway). Several functionally diverse contigs related to symbiotic bacteria including the primary endosymbiont Carsonella ruddii, Wolbachia, and CLs in the psyllid transcriptome were identified. A total of 247 contigs showed differential expression in response to CLs infection including immune and stress-related genes and vitellogenins. Expression analyses of selected psyllid genes were performed on psyllids that were exclusively reared on potato (host of the insects used for RNAseq) and psyllids exclusively reared on tomato (alternative host of psyllids). These genes showed similar expression patterns irrespective of the host plant on which the psyllids were reared, which suggests that host-plant association may not modulate expression of these genes. Our findings suggest that the impact of CLs on psyllid transcriptome was to a large extent on genes involved in metabolic processes and to a small extent on immune and stress response genes. This study is the first description of transcriptomic changes in an insect vector in response to infection with a naturally occurring bacterial plant pathogen. Data from this study provide new sequence and gene expression resources for functional genomics of potato psyllids.}, } @article {pmid22942672, year = {2012}, author = {Xia, X}, title = {DNA replication and strand asymmetry in prokaryotic and mitochondrial genomes.}, journal = {Current genomics}, volume = {13}, number = {1}, pages = {16-27}, pmid = {22942672}, issn = {1875-5488}, abstract = {Different patterns of strand asymmetry have been documented in a variety of prokaryotic genomes as well as mitochondrial genomes. Because different replication mechanisms often lead to different patterns of strand asymmetry, much can be learned of replication mechanisms by examining strand asymmetry. Here I summarize the diverse patterns of strand asymmetry among different taxonomic groups to suggest that (1) the single-origin replication may not be universal among bacterial species as the endosymbionts Wigglesworthia glossinidia, Wolbachia species, cyanobacterium Synechocystis 6803 and Mycoplasma pulmonis genomes all exhibit strand asymmetry patterns consistent with the multiple origins of replication, (2) different replication origins in some archaeal genomes leave quite different patterns of strand asymmetry, suggesting that different replication origins in the same genome may be differentially used, (3) mitochondrial genomes from representative vertebrate species share one strand asymmetry pattern consistent with the strand-displacement replication documented in mammalian mtDNA, suggesting that the mtDNA replication mechanism in mammals may be shared among all vertebrate species, and (4) mitochondrial genomes from primitive forms of metazoans such as the sponge and hydra (representing Porifera and Cnidaria, respectively), as well as those from plants, have strand asymmetry patterns similar to single-origin or multi-origin replications observed in prokaryotes and are drastically different from mitochondrial genomes from other metazoans. This may explain why sponge and hydra mitochondrial genomes, as well as plant mitochondrial genomes, evolves much slower than those from other metazoans.}, } @article {pmid22926421, year = {2013}, author = {Uni, S and Bain, O and Suzuki, K and Agatsuma, T and Harada, M and Motokawa, M and Martin, C and Lefoulon, E and Fukuda, M and Takaoka, H}, title = {Acanthocheilonema delicata n. sp. (Nematoda: Filarioidea) from Japanese badgers (Meles anakuma): description, molecular identification, and Wolbachia screening.}, journal = {Parasitology international}, volume = {62}, number = {1}, pages = {14-23}, doi = {10.1016/j.parint.2012.08.004}, pmid = {22926421}, issn = {1873-0329}, mesh = {Animals ; Electron Transport Complex IV/genetics ; Female ; Filarioidea/anatomy & histology/classification/*genetics/*microbiology/ultrastructure ; Japan ; Male ; Molecular Sequence Data ; Mustelidae/*microbiology/*parasitology ; Phylogeny ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Acanthocheilonema delicata n. sp. (Filarioidea: Onchocercidae: Onchocercinae) is described based on adult filarioids and microfilariae obtained from subcutaneous connective tissues and skin, respectively, of Japanese badgers (Meles anakuma) in Wakayama Prefecture, Japan. No endemic species of the genus had been found in Japan. Recently, some filarioids (e.g., Acanthocheilonema reconditum, Dirofilaria spp., and Onchocerca spp.) have come to light as causative agents of zoonosis worldwide. The new species was readily distinguished from its congeners by morphologic characteristics such as body length, body width, esophagus length, spicule length, and the length of microfilariae. Based on the molecular data of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene, A. delicata n. sp. was included in the clade of the genus Acanthocheilonema but differed from two other congeneric species available for study, A. viteae and A. reconditum. Acanthocheilonema delicata n. sp. did not harbor Wolbachia. It is likely that the fauna of filarioids from mammals on the Japanese islands is characterized by a high level of endemicity.}, } @article {pmid22919073, year = {2012}, author = {Darby, AC and Armstrong, SD and Bah, GS and Kaur, G and Hughes, MA and Kay, SM and Koldkjær, P and Rainbow, L and Radford, AD and Blaxter, ML and Tanya, VN and Trees, AJ and Cordaux, R and Wastling, JM and Makepeace, BL}, title = {Analysis of gene expression from the Wolbachia genome of a filarial nematode supports both metabolic and defensive roles within the symbiosis.}, journal = {Genome research}, volume = {22}, number = {12}, pages = {2467-2477}, pmid = {22919073}, issn = {1549-5469}, support = {MR/K001744/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Anti-Bacterial Agents/metabolism ; Chromatography, Liquid ; DNA Replication ; DNA, Helminth/genetics ; Female ; *Gene Expression Regulation, Bacterial ; *Genome, Bacterial ; Male ; Onchocerca volvulus/*microbiology ; Proteomics/methods ; Riboflavin/metabolism ; Sequence Analysis, RNA ; Symbiosis/*genetics ; Tandem Mass Spectrometry ; Toll-Like Receptors/genetics/metabolism ; Transcriptome ; Up-Regulation ; Wolbachia/*genetics/immunology ; }, abstract = {The α-proteobacterium Wolbachia is probably the most prevalent, vertically transmitted symbiont on Earth. In contrast with its wide distribution in arthropods, Wolbachia is restricted to one family of animal-parasitic nematodes, the Onchocercidae. This includes filarial pathogens such as Onchocerca volvulus, the cause of human onchocerciasis, or river blindness. The symbiosis between filariae and Wolbachia is obligate, although the basis of this dependency is not fully understood. Previous studies suggested that Wolbachia may provision metabolites (e.g., haem, riboflavin, and nucleotides) and/or contribute to immune defense. Importantly, Wolbachia is restricted to somatic tissues in adult male worms, whereas females also harbor bacteria in the germline. We sought to characterize the nature of the symbiosis between Wolbachia and O. ochengi, a bovine parasite representing the closest relative of O. volvulus. First, we sequenced the complete genome of Wolbachia strain wOo, which revealed an inability to synthesize riboflavin de novo. Using RNA-seq, we also generated endobacterial transcriptomes from male soma and female germline. In the soma, transcripts for membrane transport and respiration were up-regulated, while the gonad exhibited enrichment for DNA replication and translation. The most abundant Wolbachia proteins, as determined by geLC-MS, included ligands for mammalian Toll-like receptors. Enzymes involved in nucleotide synthesis were dominant among metabolism-related proteins, whereas the haem biosynthetic pathway was poorly represented. We conclude that Wolbachia may have a mitochondrion-like function in the soma, generating ATP for its host. Moreover, the abundance of immunogenic proteins in wOo suggests a role in diverting the immune system toward an ineffective antibacterial response.}, } @article {pmid22911833, year = {2012}, author = {Fu, Y and Lan, J and Zhang, Z and Hou, R and Wu, X and Yang, D and Zhang, R and Zheng, W and Nie, H and Xie, Y and Yan, N and Yang, Z and Wang, C and Luo, L and Liu, L and Gu, X and Wang, S and Peng, X and Yang, G}, title = {Novel insights into the transcriptome of Dirofilaria immitis.}, journal = {PloS one}, volume = {7}, number = {7}, pages = {e41639}, pmid = {22911833}, issn = {1932-6203}, mesh = {Animals ; Cluster Analysis ; Computational Biology ; Databases, Genetic ; Dirofilaria immitis/*genetics ; Dogs ; Expressed Sequence Tags ; Female ; Gene Expression Regulation ; Genes, Helminth/genetics ; High-Throughput Nucleotide Sequencing ; Host-Parasite Interactions/genetics ; Intestines/parasitology ; Male ; Molecular Sequence Annotation ; Molecular Sequence Data ; RNA, Messenger/genetics/metabolism ; Sequence Homology, Nucleic Acid ; Transcriptome/*genetics ; }, abstract = {BACKGROUND: The heartworm Dirofilaria immitis is the causal agent of cardiopulmonary dirofilariosis in dogs and cats, and also infects a wide range of wild mammals as well as humans. One bottleneck for the design of fundamentally new intervention and management strategies against D. immitis may be the currently limited knowledge of fundamental molecular aspects of D. immitis.

A next-generation sequencing platform combining computational approaches was employed to assess a global view of the heartworm transcriptome. A total of 20,810 unigenes (mean length  =  1,270 bp) were assembled from 22.3 million clean reads. From these, 15,698 coding sequences (CDS) were inferred, and about 85% of the unigenes had orthologs/homologs in public databases. Comparative transcriptomic study uncovered 4,157 filarial-specific genes as well as 3,795 genes potentially involved in filarial-Wolbachia symbiosis. In addition, the potential intestine transcriptome of D. immitis (1,101 genes) was mined for the first time, which might help to discover 'hidden antigens'.

CONCLUSIONS/SIGNIFICANCE: This study provides novel insights into the transcriptome of D. immitis and sheds light on its molecular processes and survival mechanisms. Furthermore, it provides a platform to discover new vaccine candidates and potential targets for new drugs against dirofilariosis.}, } @article {pmid22911683, year = {2012}, author = {Jeffery, JA and Clements, AC and Nguyen, YT and Nguyen, le H and Tran, SH and Le, NT and Vu, NS and Ryan, PA and Kay, BH}, title = {Water level flux in household containers in Vietnam--a key determinant of Aedes aegypti population dynamics.}, journal = {PloS one}, volume = {7}, number = {7}, pages = {e39067}, pmid = {22911683}, issn = {1932-6203}, mesh = {Aedes/*growth & development ; Animals ; Dengue/transmission ; Environmental Monitoring ; Family Characteristics ; Health Surveys ; Humans ; *Insect Vectors ; Population Dynamics ; Pupa ; Rural Population ; Vietnam ; Water/*parasitology ; }, abstract = {We examined changes in the abundance of immature Aedes aegypti at the household and water storage container level during the dry-season (June-July, 2008) in Tri Nguyen village, central Vietnam. We conducted quantitative immature mosquito surveys of 171 containers in the same 41 households, with replacement of samples, every two days during a 29-day period. We developed multi-level mixed effects regression models to investigate container and household variability in pupal abundance. The percentage of houses that were positive for I/II instars, III/IV instars and pupae during any one survey ranged from 19.5-43.9%, 48.8-75.6% and 17.1-53.7%, respectively. The mean numbers of Ae. aegypti pupae per house ranged between 1.9-12.6 over the study period. Estimates of absolute pupal abundance were highly variable over the 29-day period despite relatively stable weather conditions. Most variability in pupal abundance occurred at the container rather than the household level. A key determinant of Ae. aegypti production was the frequent filling of the containers with water, which caused asynchronous hatching of Ae. aegypti eggs and development of cohorts of immatures. We calculated the probability of the water volume of a large container (>500 L) increasing or decreasing by ≥20% to be 0.05 and 0.07 per day, respectively, and for small containers (<500 L) to be 0.11 and 0.13 per day, respectively. These human water-management behaviors are important determinants of Ae. aegypti production during the dry season. This has implications for choosing a suitable Wolbachia strain for release as it appears that prolonged egg desiccation does not occur in this village.}, } @article {pmid22907174, year = {2013}, author = {Sarasa, J and Bernal, A and Fernández-Calvín, B and Bella, JL}, title = {Wolbachia induced cytogenetical effects as evidenced in Chorthippus parallelus (Orthoptera).}, journal = {Cytogenetic and genome research}, volume = {139}, number = {1}, pages = {36-43}, doi = {10.1159/000341572}, pmid = {22907174}, issn = {1424-859X}, mesh = {Animals ; Cell Shape ; Chromosome Banding ; Chromosomes, Insect/genetics ; Fertility/genetics ; Genes, Bacterial ; Genetic Markers ; Grasshoppers/cytology/*genetics/microbiology ; Host-Pathogen Interactions ; Male ; Meiosis ; RNA, Ribosomal, 16S/genetics ; Spermatids/microbiology ; Symbiosis/genetics ; Wolbachia/genetics/*physiology ; X Chromosome/genetics ; }, abstract = {The cytoplasmic incompatibility induced by the bacterial endosymbiont Wolbachia is attributed to chromatin modification in the sperm of infected individuals and is only 'rescued' by infected females after fertilization. Chorthippus parallelus is a grasshopper with 2 subspecies that form a hybrid zone in the Pyrenees in which this Wolbachia-generated cytoplasmic incompatibility has recently been described. The analysis of certain cytogenetic traits (sex chromosome-linked heterochromatic bands, nucleolar organizing region expression, spermatid size and morphology, and number of chiasmata formed) in pure and hybrid Chorthippus parallelus that are infected and not infected by this bacterium indicates that the infection affects some of these traits and, in the case of the spermatids, reveals a synergism between the infection and the hybrid condition. These results are interpreted as being secondary effects of the chromatin modification induced by Wolbachia which thereby support this model of modification/rescue. The possible effects of these cytogenetic variations on affected individuals are also considered.}, } @article {pmid22903036, year = {2012}, author = {Correa, CC and Ballard, JW}, title = {Wolbachia gonadal density in female and male Drosophila vary with laboratory adaptation and respond differently to physiological and environmental challenges.}, journal = {Journal of invertebrate pathology}, volume = {111}, number = {3}, pages = {197-204}, doi = {10.1016/j.jip.2012.08.003}, pmid = {22903036}, issn = {1096-0805}, mesh = {Adaptation, Physiological ; Animals ; Bacterial Load ; Diet ; Drosophila/genetics/immunology/*microbiology/physiology ; Female ; Genetic Variation ; Male ; Ovary/*microbiology ; Sex Factors ; *Symbiosis ; Temperature ; Testis/*microbiology ; Wolbachia/*physiology ; }, abstract = {In symbiotic associations such as those between Wolbachia and insects, the within-host symbiont density plays an important role in the maintenance of the infection in natural populations, as it relates to transmission fidelity and pathogenicity of the symbiont. Within-host density is speculated to be the result of complex interactions between the bacterial genotype, the host genotype and the environment, which may account for the substantial variation in Wolbachia titres among wild collected individuals compared to laboratory lines. Using quantitative PCR, we screened the Wolbachia gonadal density of individuals from 50 isofemale Drosophila simulans lines raised in standard conditions for at least two generations after collection from the wild. Although these newly collected lines displayed significant variation of ovarian Wolbachia titres, such variation was lost by F(19). Assaying these flies at different ages and under different environmental conditions indicated that symbiont titres in female gonads were not affected by the conditions tested. However, Wolbachia density in male gonads was consistently affected by these treatments in a line-specific way. We propose that the differences in Wolbachia densities among ovaries of F(4) flies are the consequence of large differences in the field-collected females caused by the variable environment, and carried over for at least four generations. In addition, we provide evidence of sex-specific dynamics of Wolbachia in gonads of females and males. In combination, our results support the view of sex-specific Wolbachia evolutionary interactions for males and females, which has been predicted by theory and observed experimentally.}, } @article {pmid22895162, year = {2013}, author = {Jones, RT and Vetter, SM and Montenieiri, J and Holmes, J and Bernhardt, SA and Gage, KL}, title = {Yersinia pestis infection and laboratory conditions alter flea-associated bacterial communities.}, journal = {The ISME journal}, volume = {7}, number = {1}, pages = {224-228}, pmid = {22895162}, issn = {1751-7370}, mesh = {Animals ; Bartonella/*growth & development ; Flea Infestations/veterinary ; Sciuridae ; Siphonaptera/*microbiology ; Wolbachia/*growth & development ; Yersinia pestis/*physiology ; }, abstract = {We collected Oropsylla montana from rock squirrels, Spermophilus varigatus, and infected a subset of collected fleas with Yersinia pestis, the etiological agent of plague. We used bar-tagged DNA pyrosequencing to characterize bacterial communities of wild, uninfected controls and infected fleas. Bacterial communities within Y. pestis-infected fleas were substantially more similar to one another than communities within wild or control fleas, suggesting that infection alters the bacterial community in a directed manner such that specific bacterial lineages are severely reduced in abundance or entirely eliminated from the community. Laboratory conditions also significantly altered flea-associated bacterial communities relative to wild communities, but much less so than Y. pestis infection. The abundance of Firmicutes decreased considerably in infected fleas, and Bacteroidetes were almost completely eliminated from both the control and infected fleas. Bartonella and Wolbachia were unaffected or responded positively to Y. pestis infection.}, } @article {pmid22892635, year = {2012}, author = {Veneti, Z and Zabalou, S and Papafotiou, G and Paraskevopoulos, C and Pattas, S and Livadaras, I and Markakis, G and Herren, JK and Jaenike, J and Bourtzis, K}, title = {Loss of reproductive parasitism following transfer of male-killing Wolbachia to Drosophila melanogaster and Drosophila simulans.}, journal = {Heredity}, volume = {109}, number = {5}, pages = {306-312}, pmid = {22892635}, issn = {1365-2540}, mesh = {Animals ; Drosophila melanogaster ; *Evolution, Molecular ; Female ; Host-Pathogen Interactions/*genetics ; Longevity/genetics ; Male ; *Phenotype ; *Sex Characteristics ; Symbiosis/genetics ; Wolbachia/*physiology ; }, abstract = {Wolbachia manipulate insect host biology through a variety of means that result in increased production of infected females, enhancing its own transmission. A Wolbachia strain (wInn) naturally infecting Drosophila innubila induces male killing, while native strains of D. melanogaster and D. simulans usually induce cytoplasmic incompatibility (CI). In this study, we transferred wInn to D. melanogaster and D. simulans by embryonic microinjection, expecting conservation of the male-killing phenotype to the novel hosts, which are more suitable for genetic analysis. In contrast to our expectations, there was no effect on offspring sex ratio. Furthermore, no CI was observed in the transinfected flies. Overall, transinfected D. melanogaster lines displayed lower transmission rate and lower densities of Wolbachia than transinfected D. simulans lines, in which established infections were transmitted with near-perfect fidelity. In D. simulans, strain wInn had no effect on fecundity and egg-to-adult development. Surprisingly, one of the two transinfected lines tested showed increased longevity. We discuss our results in the context of host-symbiont co-evolution and the potential of symbionts to invade novel host species.}, } @article {pmid22891783, year = {2012}, author = {Wachi, N and Abe, Y and Inomata, N and Szmidt, AE and Tachida, H}, title = {Speciation history of three closely related oak gall wasps, Andricus mukaigawae, A. kashiwaphilus, and A. pseudoflos (Hymenoptera: Cynipidae) inferred from nuclear and mitochondrial DNA sequences.}, journal = {Molecular ecology}, volume = {21}, number = {19}, pages = {4681-4694}, doi = {10.1111/j.1365-294X.2012.05727.x}, pmid = {22891783}, issn = {1365-294X}, mesh = {Animals ; Bayes Theorem ; Cell Nucleus/*genetics ; DNA, Mitochondrial/*genetics ; Gene Flow ; *Genetic Speciation ; Genetic Variation ; Likelihood Functions ; Models, Genetic ; Molecular Sequence Data ; Parthenogenesis ; Phylogeny ; Quercus/*parasitology ; Sequence Analysis, DNA ; Wasps/*classification/genetics/microbiology ; Wolbachia ; }, abstract = {The Andricus mukaigawae complex of oak gall wasps is composed of cyclically parthenogenetic species: A. mukaigawae and Andricus kashiwaphilus, and a parthenogenetic species, Andricus pseudoflos. The component species differ in life history, host plant, karyotype, and asexual gall shape, although little difference is found in the external morphology of asexual adults. To understand the speciation history of this species complex, DNA sequences of one mitochondrial region and nine nuclear gene regions were investigated. The genetic relationship among the species suggested that a loss of sex occurred after host shift. Unexpectedly, two or three distinct groups in the parthenogenetic species, A. pseudoflos, were revealed by both mitochondrial and nuclear DNA data. Gene flow in nuclear genes from the species not infected by Wolbachia (A. kashiwaphilus) to the species infected by it (A. mukaigawae) was suggested by a method based on coalescent simulations. On the other hand, gene flow in mitochondrial genes was suggested to be in the opposite direction. These findings indicate possible involvement of Wolbachia infection in the speciation process of the A. mukaigawae complex.}, } @article {pmid22889830, year = {2012}, author = {Godel, C and Kumar, S and Koutsovoulos, G and Ludin, P and Nilsson, D and Comandatore, F and Wrobel, N and Thompson, M and Schmid, CD and Goto, S and Bringaud, F and Wolstenholme, A and Bandi, C and Epe, C and Kaminsky, R and Blaxter, M and Mäser, P}, title = {The genome of the heartworm, Dirofilaria immitis, reveals drug and vaccine targets.}, journal = {FASEB journal : official publication of the Federation of American Societies for Experimental Biology}, volume = {26}, number = {11}, pages = {4650-4661}, pmid = {22889830}, issn = {1530-6860}, support = {095831//Wellcome Trust/United Kingdom ; G0900740/MRC_/Medical Research Council/United Kingdom ; /BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Anthelmintics/*pharmacology/therapeutic use ; Dirofilaria immitis/drug effects/*genetics/immunology/microbiology ; Dirofilariasis/drug therapy/*parasitology/prevention & control ; Dog Diseases/drug therapy/*parasitology/prevention & control ; Dogs ; Female ; Genetic Variation ; Genome, Bacterial ; *Genome, Helminth ; Male ; Phylogeny ; Proteome ; RNA, Helminth/chemistry ; Symbiosis ; Transcriptome/genetics ; Vaccines/*immunology ; Wolbachia/genetics/physiology ; }, abstract = {The heartworm Dirofilaria immitis is an important parasite of dogs. Transmitted by mosquitoes in warmer climatic zones, it is spreading across southern Europe and the Americas at an alarming pace. There is no vaccine, and chemotherapy is prone to complications. To learn more about this parasite, we have sequenced the genomes of D. immitis and its endosymbiont Wolbachia. We predict 10,179 protein coding genes in the 84.2 Mb of the nuclear genome, and 823 genes in the 0.9-Mb Wolbachia genome. The D. immitis genome harbors neither DNA transposons nor active retrotransposons, and there is very little genetic variation between two sequenced isolates from Europe and the United States. The differential presence of anabolic pathways such as heme and nucleotide biosynthesis hints at the intricate metabolic interrelationship between the heartworm and Wolbachia. Comparing the proteome of D. immitis with other nematodes and with mammalian hosts, we identify families of potential drug targets, immune modulators, and vaccine candidates. This genome sequence will support the development of new tools against dirofilariasis and aid efforts to combat related human pathogens, the causative agents of lymphatic filariasis and river blindness.}, } @article {pmid22878693, year = {2012}, author = {Bennett, GM and Pantoja, NA and O'Grady, PM}, title = {Diversity and phylogenetic relationships of Wolbachia in Drosophila and other native Hawaiian insects.}, journal = {Fly}, volume = {6}, number = {4}, pages = {273-283}, pmid = {22878693}, issn = {1933-6942}, mesh = {Alleles ; Animals ; Drosophila/genetics/*microbiology ; Hawaii ; Insecta/classification/genetics/microbiology ; Likelihood Functions ; *Phylogeny ; Species Specificity ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Wolbachia is a genus of parasitic alphaproteobacteria found in arthropods and nematodes, and represents on of the most common, widespread endosymbionts known. Wolbachia affects a variety of reproductive functions in its host (e.g., male killing, cytoplasmic incompatibility, parthenogenesis), which have the potential to dramatically impact host evolution and species formation. Here, we present the first broad-scale study to screen natural populations of native Hawaiian insects for Wolbachia, focusing on the endemic Diptera. Results indicate that Wolbachia infects native Hawaiian taxa, with alleles spanning phylogenetic supergroups, A and B. The overall frequency of Wolbachia incidene in Hawaiian insects was 14%. The incidence of infection in native Hawaiian Diptera was 11% for individuals and 12% for all species screened. Wolbachia was not detected in two large, widespread Hawaiian dipteran families-Dolichopodidae (44 spp screened) and Limoniidae (12 spp screened). Incidence of infection within endemic Hawaiian lineages that carry Wolbachia was 18% in Drosophilidae species, 25% in Caliphoridae species, > 90% in Nesophrosyne species, 20% in Drosophila dasycnemia and 100% in Nesophrosyne craterigena. Twenty unique alleles were recovered in this study, of which 18 are newly recorded. Screening of endemic populations of D. dasycnemia across Hawaii Island revealed 4 unique alleles. Phylogenetic relationships and allele diversity provide evidence for horizontal transfer of Wolbachia among Hawaiian arthropod lineages.}, } @article {pmid22876183, year = {2012}, author = {Le Clec'h, W and Braquart-Varnier, C and Raimond, M and Ferdy, JB and Bouchon, D and Sicard, M}, title = {High virulence of Wolbachia after host switching: when autophagy hurts.}, journal = {PLoS pathogens}, volume = {8}, number = {8}, pages = {e1002844}, pmid = {22876183}, issn = {1553-7374}, mesh = {Animals ; *Autophagy ; Central Nervous System/metabolism/microbiology/ultrastructure ; *Host-Pathogen Interactions ; Isopoda ; Rickettsiaceae Infections/*metabolism/microbiology ; Species Specificity ; Vacuoles/metabolism/microbiology/ultrastructure ; Wolbachia/*pathogenicity/*physiology/ultrastructure ; }, abstract = {Wolbachia are widespread endosymbionts found in a large variety of arthropods. While these bacteria are generally transmitted vertically and exhibit weak virulence in their native hosts, a growing number of studies suggests that horizontal transfers of Wolbachia to new host species also occur frequently in nature. In transfer situations, virulence variations can be predicted since hosts and symbionts are not adapted to each other. Here, we describe a situation where a Wolbachia strain (wVulC) becomes a pathogen when transfected from its native terrestrial isopod host species (Armadillidium vulgare) to another species (Porcellio d. dilatatus). Such transfer of wVulC kills all recipient animals within 75 days. Before death, animals suffer symptoms such as growth slowdown and nervous system disorders. Neither those symptoms nor mortalities were observed after injection of wVulC into its native host A. vulgare. Analyses of wVulC's densities in main organs including Central Nervous System (CNS) of both naturally infected A. vulgare and transfected P. d. dilatatus and A. vulgare individuals revealed a similar pattern of host colonization suggesting an overall similar resistance of both host species towards this bacterium. However, for only P. d. dilatatus, we observed drastic accumulations of autophagic vesicles and vacuoles in the nerve cells and adipocytes of the CNS from individuals infected by wVulC. The symptoms and mortalities could therefore be explained by this huge autophagic response against wVulC in P. d. dilatatus cells that is not triggered in A. vulgare. Our results show that Wolbachia (wVulC) can lead to a pathogenic interaction when transferred horizontally into species that are phylogenetically close to their native hosts. This change in virulence likely results from the autophagic response of the host, strongly altering its tolerance to the symbiont and turning it into a deadly pathogen.}, } @article {pmid22874746, year = {2013}, author = {Wang, J and Brelsfoard, C and Wu, Y and Aksoy, S}, title = {Intercommunity effects on microbiome and GpSGHV density regulation in tsetse flies.}, journal = {Journal of invertebrate pathology}, volume = {112 Suppl}, number = {0}, pages = {S32-9}, pmid = {22874746}, issn = {1096-0805}, support = {R01 GM069449/GM/NIGMS NIH HHS/United States ; AI051584/AI/NIAID NIH HHS/United States ; R03 TW008755/TW/FIC NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; AI068932/AI/NIAID NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Infections/transmission ; DNA Viruses ; Female ; *Insect Viruses ; Male ; *Metagenome ; Symbiosis ; Tsetse Flies/*microbiology ; Wigglesworthia ; Wolbachia ; }, abstract = {Tsetse flies have a highly regulated and defined microbial fauna made of 3 bacterial symbionts (obligate Wigglesworthia glossinidia, commensal Sodalis glossinidius and parasitic Wolbachia pipientis) in addition to a DNA virus (Glossina pallidipes Salivary gland Hypertrophy Virus, GpSGHV). It has been possible to rear flies in the absence of either Wigglesworthia or in totally aposymbiotic state by dietary supplementation of tsetse's bloodmeal. In the absence of Wigglesworthia, tsetse females are sterile, and adult progeny are immune compromised. The functional contributions for Sodalist are less known, while Wolbachia cause reproductive manupulations known as cytoplasmic incompatibility (CI). High GpSGHV virus titers result in reduced fecundity and lifespan, and have compromised efforts to colonize flies in the insectary for large rearing purposes. Here we investigated the within community effects on the density regulation of the individual microbiome partners in tsetse lines with different symbiotic compositions. We show that absence of Wigglesworthia results in loss of Sodalis in subsequent generations possibly due to nutritional dependancies between the symbiotic partners. While an initial decrease in Wolbachia and GpSGHV levels are also noted in the absence of Wigglesworthia, these infections eventually reach homeostatic levels indicating adaptations to the new host immune environment or nutritional ecology. Absence of all bacterial symbionts also results in an initial reduction of viral titers, which recover in the second generation. Our findings suggest that in addition to the host immune system, interdependencies between symbiotic partners result in a highly tuned density regulation for tsetse's microbiome.}, } @article {pmid22851652, year = {2012}, author = {Chen, S and Oliveira, MT and Sanz, A and Kemppainen, E and Fukuoh, A and Schlicht, B and Kaguni, LS and Jacobs, HT}, title = {A cytoplasmic suppressor of a nuclear mutation affecting mitochondrial functions in Drosophila.}, journal = {Genetics}, volume = {192}, number = {2}, pages = {483-493}, pmid = {22851652}, issn = {1943-2631}, support = {R01 GM045295/GM/NIGMS NIH HHS/United States ; GM45295/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cell Nucleus/genetics/metabolism ; DNA Copy Number Variations ; DNA Replication/*genetics ; *DNA, Mitochondrial/genetics/metabolism ; Drosophila Proteins/genetics/metabolism ; *Drosophila melanogaster/genetics/metabolism/microbiology ; Evolution, Molecular ; Humans ; Mitochondrial Diseases/genetics/metabolism ; Mutation ; *Oxidative Phosphorylation ; Phenotype ; Positive Transcriptional Elongation Factor B/genetics/metabolism ; RNA, Untranslated/genetics ; Wolbachia/genetics/pathogenicity ; }, abstract = {Phenotypes relevant to oxidative phosphorylation (OXPHOS) in eukaryotes are jointly determined by nuclear and mitochondrial DNA (mtDNA). Thus, in humans, the variable clinical presentations of mitochondrial disease patients bearing the same primary mutation, whether in nuclear or mitochondrial DNA, have been attributed to putative genetic determinants carried in the "other" genome, though their identity and the molecular mechanism(s) by which they might act remain elusive. Here we demonstrate cytoplasmic suppression of the mitochondrial disease-like phenotype of the Drosophila melanogaster nuclear mutant tko(25t), which includes developmental delay, seizure sensitivity, and defective male courtship. The tko(25t) strain carries a mutation in a mitoribosomal protein gene, causing OXPHOS deficiency due to defective intramitochondrial protein synthesis. Phenotypic suppression was associated with increased mtDNA copy number and increased mitochondrial biogenesis, as measured by the expression levels of porin voltage dependent anion channel and Spargel (PGC1α). Ubiquitous overexpression of Spargel in tko(25t) flies phenocopied the suppressor, identifying it as a key mechanistic target thereof. Suppressor-strain mtDNAs differed from related nonsuppressor strain mtDNAs by several coding-region polymorphisms and by length and sequence variation in the noncoding region (NCR), in which the origin of mtDNA replication is located. Cytoplasm from four of five originally Wolbachia-infected strains showed the same suppressor effect, whereas that from neither of two uninfected strains did so, suggesting that the stress of chronic Wolbachia infection may provide evolutionary selection for improved mitochondrial fitness under metabolic stress. Our findings provide a paradigm for understanding the role of mtDNA genotype in human disease.}, } @article {pmid22848774, year = {2012}, author = {Lu, P and Bian, G and Pan, X and Xi, Z}, title = {Wolbachia induces density-dependent inhibition to dengue virus in mosquito cells.}, journal = {PLoS neglected tropical diseases}, volume = {6}, number = {7}, pages = {e1754}, pmid = {22848774}, issn = {1935-2735}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; R01AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*virology ; Animal Structures/microbiology ; Animals ; *Antibiosis ; Bacterial Load ; Cell Line ; Dengue Virus/*growth & development ; Wolbachia/*growth & development ; }, abstract = {Wolbachia is a maternal transmitted endosymbiotic bacterium that is estimated to infect up to 65% of insect species. The ability of Wolbachia to both induce viral interference and spread into mosquito vector population makes it possible to develop Wolbachia as a biological control agent for dengue control. While Wolbachia induces resistance to dengue virus in the transinfected Aedes aegypti mosquitoes, a similar effect was not observed in Aedes albopictus, which naturally carries Wolbachia infection but still serves as a dengue vector. In order to understand the mechanism of this lack of Wolbachia-mediated viral interference, we used both Ae. albopictus cell line (Aa23) and mosquitoes to characterize the impact of Wolbachia on dengue infection. A serial of sub-lethal doses of antibiotic treatment was used to partially remove Wolbachia in Aa23 cells and generate cell cultures with Wolbachia at different densities. We show that there is a strong negative linear correlation between the genome copy of Wolbachia and dengue virus with a dengue infection completely removed when Wolbacha density reaches a certain level. We then compared Wolbachia density between transinfected Ae. aegypti and naturally infected Ae. albopictus. The results show that Wolbachia density in midgut, fatbody and salivary gland of Ae. albopictus is 80-, 18-, and 24-fold less than that of Ae. aegypti, respectively. We provide evidence that Wolbachia density in somatic tissues of Ae. albopictus is too low to induce resistance to dengue virus. Our results will aid in understanding the mechanism of Wolbachia-mediated pathogen interference and developing novel methods to block disease transmission by mosquitoes carrying native Wolbachia infections.}, } @article {pmid22844425, year = {2012}, author = {Kodandaramaiah, U and Weingartner, E and Janz, N and Leski, M and Slove, J and Warren, A and Nylin, S}, title = {Investigating concordance among genetic data, subspecies circumscriptions and hostplant use in the nymphalid butterfly Polygonia faunus.}, journal = {PloS one}, volume = {7}, number = {7}, pages = {e41058}, pmid = {22844425}, issn = {1932-6203}, mesh = {Animals ; Butterflies/*classification/*genetics/microbiology ; Cluster Analysis ; DNA, Mitochondrial/genetics ; Genetic Variation/genetics ; *Magnoliopsida ; Microsatellite Repeats/genetics ; Phylogeography ; Species Specificity ; Wolbachia/physiology ; }, abstract = {Subspecies are commonly used taxonomic units to formally describe intraspecific geographic variation in morphological traits. However, the concept of subspecies is not clearly defined, and there is little agreement about what they represent in terms of evolutionary units, and whether they can be used as reliably useful units in conservation, evolutionary theory and taxonomy. We here investigate whether the morphologically well-characterized subspecies in the North American butterfly Polygonia faunus are supported by genetic data from mitochondrial sequences and eight microsatellite loci. We also investigate the phylogeographic structure of P. faunus and test whether similarities in host-plant use among populations are related to genetic similarity. Neither the nuclear nor the mitochondrial data corroborated subspecies groupings. We found three well defined genetic clusters corresponding to California, Arizona and (New Mexico+Colorado). There was little structuring among the remaining populations, probably due to gene flow across populations. We found no support for the hypothesis that similarities in host use are related to genetic proximity. The results indicate that the species underwent a recent rapid expansion, probably from two glacial refugia in western North America. The mitochondrial haplotype network indicates at least two independent expansion phases into eastern North America. Our results clearly demonstrate that subspecies in P. faunus do not conform to the structuring of genetic variation. More studies on insects and other invertebrates are needed to better understand the scope of this phenomenon. The results of this study will be crucial in designing further experiments to understand the evolution of hostplant utilization in this species.}, } @article {pmid22843518, year = {2012}, author = {Osborne, SE and Iturbe-Ormaetxe, I and Brownlie, JC and O'Neill, SL and Johnson, KN}, title = {Antiviral protection and the importance of Wolbachia density and tissue tropism in Drosophila simulans.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {19}, pages = {6922-6929}, pmid = {22843518}, issn = {1098-5336}, mesh = {Animal Structures/microbiology ; Animals ; Anti-Bacterial Agents/administration & dosage ; Bacterial Load ; Dengue Virus/*growth & development/immunology ; Drosophila/*microbiology/*virology ; Tetracycline/administration & dosage ; Wolbachia/drug effects/*growth & development/isolation & purification ; }, abstract = {Wolbachia, a maternally transmitted endosymbiont of insects, is increasingly being seen as an effective biological control agent that can interfere with transmission of pathogens, including dengue virus. However, the mechanism of antiviral protection is not well understood. The density and distribution of Wolbachia in host tissues have been implicated as contributing factors by previous studies with both mosquitoes and flies. Drosophila flies infected with five diverse strains of Wolbachia were screened for the ability to mediate antiviral protection. The three protective Wolbachia strains were more closely related and occurred at a higher density within whole flies than the two nonprotective Wolbachia strains. In this study, to further investigate the relationship between whole-fly Wolbachia density and the ability to mediate antiviral protection, tetracycline was used to decrease the abundance of the high-density, protective Wolbachia strain wAu prior to viral challenge. Antiviral protection was lost when the density of the protective Wolbachia strain was decreased to an abundance similar to that of nonprotective Wolbachia strains. We determined the Wolbachia density and distribution in tissues of the same five fly-Wolbachia combinations as used previously. The Wolbachia density within the head, gut, and Malpighian tubules correlated with the ability to mediate antiviral protection. These findings may facilitate the development of Wolbachia biological control strategies and help to predict host-Wolbachia pairings that may interfere with virus-induced pathology.}, } @article {pmid22841948, year = {2013}, author = {Soumana, IH and Simo, G and Njiokou, F and Tchicaya, B and Abd-Alla, AM and Cuny, G and Geiger, A}, title = {The bacterial flora of tsetse fly midgut and its effect on trypanosome transmission.}, journal = {Journal of invertebrate pathology}, volume = {112 Suppl}, number = {}, pages = {S89-93}, doi = {10.1016/j.jip.2012.03.029}, pmid = {22841948}, issn = {1096-0805}, mesh = {Animals ; Cattle ; Humans ; Insect Vectors/microbiology ; Intestines/*microbiology ; Symbiosis ; Trypanosoma/parasitology ; Trypanosomiasis, African/microbiology/*transmission ; Tsetse Flies/*microbiology ; }, abstract = {The tsetse fly, Glossina palpalis is a vector of the trypanosome that causes sleeping sickness in humans and nagana in cattle along with associated human health problems and massive economic losses. The insect is also known to carry a number of symbionts such as Sodalis, Wigglesworthia, Wolbachia whose effects on the physiology of the insect have been studied in depth. However, effects of other bacterial flora on the physiology of the host and vector competence have received little attention. Epidemiological studies on tsetse fly populations from different geographic sites revealed the presence of a variety of bacteria in the midgut. The most common of the flora belong to the genera Entrobacter (most common), Enterococcus, and Acinetobacter. It was a little surprising to find such diversity in the tsetse midgut since the insect is monophagous consuming vertebrate blood only. Diversity of bacteria is normally associated with polyphagous insects. In contrast to the symbionts, the role of resident midgut bacterial flora on the physiology of the fly and vector competence remains to be elucidated. With regard, Sodalis glossinidius, our data showed that flies harbouring this symbiont have three times greater probability of being infected by trypanosomes than flies without the symbiont. The data delineated in these studies under score the need to carry out detailed investigations on the role of resident bacteria on the physiology of the fly and vector competence.}, } @article {pmid22837817, year = {2012}, author = {Duncan, AB and Agnew, P and Noel, V and Demettre, E and Seveno, M and Brizard, JP and Michalakis, Y}, title = {Proteome of Aedes aegypti in response to infection and coinfection with microsporidian parasites.}, journal = {Ecology and evolution}, volume = {2}, number = {4}, pages = {681-694}, pmid = {22837817}, issn = {2045-7758}, abstract = {Hosts are frequently infected with more than one parasite or pathogen at any one time, but little is known as to how they respond to multiple immune challenges compared to those involving single infections. We investigated the proteome of Aedes aegypti larvae following infection with either Edhazardia aedis or Vavraia culicis, and coinfections involving both. They are both obligate intracellular parasites belonging to the phylum microsporidia and infect natural populations of Ae. aegypti. The results found some proteins only showing modified abundance in response to infections involving E. aedis, while others were only differentially abundant when infections involved V. culicis. Some proteins only responded with modified abundance to the coinfection condition, while others were differentially abundant in response to all three types of infection. As time since infection increased, the response to each of the single parasite infections diverged, while the response to the E. aedis and coinfection treatments converged. Some of the proteins differentially abundant in response to infection were identified. They included two vacuolar ATPases, proteins known to have a role in determining the infection success of intracellular parasites. This result suggests microsporidia could influence the infection success of other intracellular pathogens infecting vector species of mosquito, including viruses, Plasmodium and Wolbachia.}, } @article {pmid22835476, year = {2013}, author = {Doudoumis, V and Alam, U and Aksoy, E and Abd-Alla, AM and Tsiamis, G and Brelsfoard, C and Aksoy, S and Bourtzis, K}, title = {Tsetse-Wolbachia symbiosis: comes of age and has great potential for pest and disease control.}, journal = {Journal of invertebrate pathology}, volume = {112 Suppl}, number = {0}, pages = {S94-103}, pmid = {22835476}, issn = {1096-0805}, support = {AI06892/AI/NIAID NIH HHS/United States ; R03TW008413/TW/FIC NIH HHS/United States ; R03 TW008755/TW/FIC NIH HHS/United States ; D43 TW007391/TW/FIC NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; D43TW007391/TW/FIC NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; R03 TW008413/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; Gene Transfer, Horizontal/genetics ; Humans ; Pest Control, Biological/*methods ; *Symbiosis/genetics ; Trypanosomiasis, African/prevention & control ; Tsetse Flies/genetics/*microbiology ; *Wolbachia/genetics ; }, abstract = {Tsetse flies (Diptera: Glossinidae) are the sole vectors of African trypanosomes, the causative agent of sleeping sickness in human and nagana in animals. Like most eukaryotic organisms, Glossina species have established symbiotic associations with bacteria. Three main symbiotic bacteria have been found in tsetse flies: Wigglesworthia glossinidia, an obligate symbiotic bacterium, the secondary endosymbiont Sodalis glossinidius and the reproductive symbiont Wolbachia pipientis. In the present review, we discuss recent studies on the detection and characterization of Wolbachia infections in Glossina species, the horizontal transfer of Wolbachia genes to tsetse chromosomes, the ability of this symbiont to induce cytoplasmic incompatibility in Glossina morsitans morsitans and also how new environment-friendly tools for disease control could be developed by harnessing Wolbachia symbiosis.}, } @article {pmid22835046, year = {2012}, author = {Crespi, B and Schwander, T}, title = {Asexual evolution: do intragenomic parasites maintain sex?.}, journal = {Molecular ecology}, volume = {21}, number = {16}, pages = {3893-3895}, doi = {10.1111/j.1365-294X.2012.05638.x}, pmid = {22835046}, issn = {1365-294X}, mesh = {Animals ; Female ; Reproduction, Asexual/*genetics ; Wasps/*physiology ; Wolbachia/*physiology ; }, abstract = {Resolving the paradox of sex, with its twofold cost to genic transmission, remains one of the major unresolved questions in evolutionary biology. Counting this genetic cost has now gone genomic. In this issue of Molecular Ecology, Kraaijeveld et al. (2012) describe the first genome-scale comparative study of related sexual and asexual animal lineages, to test the hypothesis that asexuals bear heavier loads of deleterious transposable elements. A much higher density of such parasites might be expected, due to the inability of asexual lineages to purge transposons via mechanisms exclusive to sexual reproduction. They find that the answer is yes--and no--depending upon the family of transposons considered. Like many such advances in testing theory, more questions are raised by this study than answered, but a door has been opened to molecular evolutionary analyses of how responses to selection from intragenomic parasites might mediate the costs of sex.}, } @article {pmid22831171, year = {2012}, author = {Brennan, LJ and Haukedal, JA and Earle, JC and Keddie, B and Harris, HL}, title = {Disruption of redox homeostasis leads to oxidative DNA damage in spermatocytes of Wolbachia-infected Drosophila simulans.}, journal = {Insect molecular biology}, volume = {21}, number = {5}, pages = {510-520}, doi = {10.1111/j.1365-2583.2012.01155.x}, pmid = {22831171}, issn = {1365-2583}, mesh = {8-Hydroxy-2'-Deoxyguanosine ; Animals ; Antioxidants ; Cell Line ; Comet Assay ; *DNA Damage ; Deoxyguanosine/analogs & derivatives/analysis ; Drosophila/metabolism/*microbiology ; Female ; Glutathione ; Homeostasis ; Male ; *Oxidative Stress ; Spermatocytes/metabolism ; *Spermatogenesis ; Superoxide Dismutase/metabolism ; Testis/metabolism/microbiology ; Wolbachia/*physiology ; }, abstract = {Molecular interactions between symbiotic bacteria and their animal hosts are, as yet, poorly understood. The most widespread bacterial endosymbiont, Wolbachia pipientis, occurs in high density in testes of infected Drosophila simulans and causes cytoplasmic incompatibility (CI), a form of male-derived zygotic lethality. Wolbachia grow and divide within host vacuoles that generate reactive oxygen species (ROS), which in turn stimulate the up-regulation of antioxidant enzymes. These enzymes appear to protect the host from ROS-mediated damage, as there is no obvious fitness cost to Drosophila carrying Wolbachia infections. We have now determined that DNA from Wolbachia-infected mosquito Aedes albopictus (Aa23) cells shows a higher amount of the base 8-oxo-deoxyguanosine, a marker of oxidative DNA damage, than DNA from uninfected cells, and that Wolbachia infection in D. simulans is associated with an increase in DNA strand breaks in meiotic spermatocytes. Feeding exogenous antioxidants to male and female D. simulans dramatically increased Wolbachia numbers with no obvious effects on host fitness. These results suggest that ROS-induced DNA damage in sperm nuclei may contribute to the modification characteristic of CI expression in Wolbachia-infected males and that Wolbachia density is sensitive to redox balance in these flies.}, } @article {pmid22825287, year = {2012}, author = {Chen, LH and Wilson, ME}, title = {Dengue and chikungunya in travelers: recent updates.}, journal = {Current opinion in infectious diseases}, volume = {25}, number = {5}, pages = {523-529}, doi = {10.1097/QCO.0b013e328356ffd5}, pmid = {22825287}, issn = {1473-6527}, mesh = {*Alphavirus Infections/drug therapy/epidemiology/transmission ; Animals ; Antiviral Agents/therapeutic use ; Chikungunya Fever ; *Dengue/diagnosis/epidemiology/prevention & control/transmission ; Disease Outbreaks/prevention & control ; Disease Vectors ; Humans ; Mosquito Control/methods ; *Travel ; }, abstract = {PURPOSE OF REVIEW: Dengue virus (DENV) and chikungunya virus (CHIKV) cause significant morbidity in many world regions and their epidemiology, pathogenesis, vector issues, and control and prevention continue to fascinate researchers. This review focuses on the progress in these areas in the past 2 years.

RECENT FINDINGS: Recent studies have highlighted the features of the epidemiology of DENV and CHIKV in Africa, some places of transmission being detected through travelers as sentinels. Autochthonous transmission of both viruses has been documented in France, raising concern regarding the potential for outbreaks where a competent vector, Aedes albopictus, is present. Some aspects of immune response following DENV and CHIKV infections have been elucidated. New diagnostic techniques for DENV have been explored as well as treatment for CHIKV. Research on Wolbachia as a means of biologic control of mosquito-borne infections including DENV and CHIKV has made significant progress.

SUMMARY: These studies further our understanding of the evolving DENV and CHIKV epidemiology and potential transmission in nonendemic regions, and can contribute to the assessment of travelers. The new information on immunological responses to these viruses influences the vaccine development. Risk factors for severe disease and new therapeutic options could improve current treatment. Finally, research on Wolbachia holds promise in these two important vector-borne viral infections.}, } @article {pmid22824741, year = {2013}, author = {Merkling, SH and van Rij, RP}, title = {Beyond RNAi: antiviral defense strategies in Drosophila and mosquito.}, journal = {Journal of insect physiology}, volume = {59}, number = {2}, pages = {159-170}, doi = {10.1016/j.jinsphys.2012.07.004}, pmid = {22824741}, issn = {1879-1611}, mesh = {Animals ; Arboviruses/physiology ; Culicidae/genetics/*immunology/virology ; Drosophila melanogaster/genetics/*immunology/virology ; Evolution, Molecular ; *Immunity, Innate ; Insect Proteins/genetics/*metabolism ; RNA Interference ; Signal Transduction ; }, abstract = {Virus transmission and spread by arthropods is a major economic and public health concern. The ongoing dissemination of arthropod-borne viruses by blood-feeding insects is an important incentive to study antiviral immunity in these animals. RNA interference is a major mechanism for antiviral defense in insects, including the fruit fly Drosophila melanogaster and several vector mosquitoes. However, recent data suggest that the evolutionary conserved Toll, Imd and Jak-Stat signaling pathways also contribute to antiviral immunity. Moreover, symbionts, such as the intracellular bacterium Wolbachia and the gut microflora, influence the course of virus infection in insects. These results add an additional level of complexity to antiviral immunity, but also provide novel opportunities to control the spread of arboviruses. In this review, we provide an overview of the current knowledge and recent developments in antiviral immunity in Dipteran insects, with a focus on non-RNAi mediated inducible responses.}, } @article {pmid22815933, year = {2012}, author = {Shrivastava, N and Nag, JK and Misra-Bhattacharya, S}, title = {Molecular characterization of NAD+-dependent DNA ligase from Wolbachia endosymbiont of lymphatic filarial parasite Brugia malayi.}, journal = {PloS one}, volume = {7}, number = {7}, pages = {e41113}, pmid = {22815933}, issn = {1932-6203}, mesh = {Amino Acid Sequence ; Animals ; Blotting, Western ; Brugia malayi/metabolism ; Catalysis ; Circular Dichroism ; DNA Ligases/*chemistry ; Female ; Immunoblotting ; Male ; Mice ; Mice, Inbred BALB C ; Microscopy, Confocal/methods ; Molecular Sequence Data ; Phylogeny ; Recombinant Proteins/metabolism ; Sequence Homology, Amino Acid ; Spectrometry, Fluorescence/methods ; Wolbachia/*metabolism ; }, abstract = {The lymphatic filarial parasite, Brugia malayi contains Wolbachia endobacteria that are essential for development, viability and fertility of the parasite. Therefore, wolbachial proteins have been currently seen as the potential antifilarial drug targets. NAD(+)-dependent DNA ligase is characterized as a promising drug target in several organisms due to its crucial, indispensable role in DNA replication, recombination and DNA repair. We report here the cloning, expression and purification of NAD(+)-dependent DNA ligase of Wolbachia endosymbiont of B. malayi (wBm-LigA) for its molecular characterization. wBm-LigA has all the domains that are present in nearly all the eubacterial NAD(+)-dependent DNA ligases such as N-terminal adenylation domain, OB fold, helix-hairpin-helix (HhH) and BRCT domain except zinc-binding tetracysteine domain. The purified recombinant protein (683-amino acid) was found to be biochemically active and was present in its native form as revealed by the circular dichroism and fluorescence spectra. The purified recombinant enzyme was able to catalyze intramolecular strand joining on a nicked DNA as well as intermolecular joining of the cohesive ends of BstEII restricted lamda DNA in an in vitro assay. The enzyme was localized in the various life-stages of B. malayi parasites by immunoblotting and high enzyme expression was observed in Wolbachia within B. malayi microfilariae and female adult parasites along the hypodermal chords and in the gravid portion as evident by the confocal microscopy. Ours is the first report on this enzyme of Wolbachia and these findings would assist in validating the antifilarial drug target potential of wBm-LigA in future studies.}, } @article {pmid22808174, year = {2012}, author = {Rottschaefer, SM and Lazzaro, BP}, title = {No effect of Wolbachia on resistance to intracellular infection by pathogenic bacteria in Drosophila melanogaster.}, journal = {PloS one}, volume = {7}, number = {7}, pages = {e40500}, pmid = {22808174}, issn = {1932-6203}, support = {R01 AI083932/AI/NIAID NIH HHS/United States ; }, mesh = {Analysis of Variance ; Animals ; Bacterial Load/drug effects ; Disease Resistance/drug effects/*immunology ; Drosophila melanogaster/drug effects/*immunology/*microbiology ; Enterobacteriaceae Infections/*immunology/*microbiology ; Genotype ; Intracellular Space/drug effects/*microbiology ; Listeria monocytogenes/drug effects ; Providencia/drug effects/physiology ; Salmonella typhimurium/drug effects ; Tetracycline/pharmacology ; Wolbachia/drug effects/genetics/*physiology ; }, abstract = {Multiple studies have shown that infection with the endosymbiotic bacterium Wolbachia pipientis confers Drosophila melanogaster and other insects with resistance to infection by RNA viruses. Studies investigating whether Wolbachia infection induces the immune system or confers protection against secondary bacterial infection have not shown any effect. These studies, however, have emphasized resistance against extracellular pathogens. Since Wolbachia lives inside the host cell, we hypothesized that Wolbachia might confer resistance to pathogens that establish infection by invading host cells. We therefore tested whether Wolbachia-infected D. melanogaster are protected against infection by the intracellular pathogenic bacteria Listeria monocytogenes and Salmonella typhimurium, as well as the extracellular pathogenic bacterium Providencia rettgeri. We evaluated the ability of flies infected with Wolbachia to suppress secondary infection by pathogenic bacteria relative to genetically matched controls that had been cured of Wolbachia by treatment with tetracycline. We found no evidence that Wolbachia alters host ability to suppress proliferation of any of the three pathogenic bacteria. Our results indicate that Wolbachia-induced antiviral protection does not result from a generalized response to intracellular pathogens.}, } @article {pmid22806335, year = {2012}, author = {Zhu, LY and Zhang, KJ and Zhang, YK and Ge, C and Gotoh, T and Hong, XY}, title = {Wolbachia strengthens cardinium-induced cytoplasmic incompatibility in the spider mite Tetranychus piercei McGregor.}, journal = {Current microbiology}, volume = {65}, number = {5}, pages = {516-523}, pmid = {22806335}, issn = {1432-0991}, mesh = {Animals ; Bacteroidetes/genetics/*physiology ; Cytoplasm/genetics/*microbiology/physiology ; Female ; Male ; Reproduction ; Symbiosis ; Tetranychidae/genetics/*microbiology/*physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia and Cardinium are maternally inherited intracellular bacteria that can manipulate the reproduction of their arthropod hosts, such as by inducing cytoplasmic incompatibility (CI). Although the reproductive alteration induced by Wolbachia or Cardinium have been well investigated, the effects of these two endosymbionts co-infecting the same host are poorly understood. We found that Tetranychus piercei McGregor is naturally infected with Wolbachia and Cardinium. We performed all possible crossing combinations using naturally infected and cured strains, and the results show that Wolbachia induced a weak level of CI, while Cardinium-infected and doubly infected males caused severe CI. Wolbachia and Cardinium could not rescue CI each other; however, Wolbachia boosted the expression of Cardinium-induced CI. Quantitative PCR results demonstrated that CI was associated with the infection density of Wolbachia and Cardinium.}, } @article {pmid22798369, year = {2012}, author = {Hedges, LM and Yamada, R and O'Neill, SL and Johnson, KN}, title = {The small interfering RNA pathway is not essential for Wolbachia-mediated antiviral protection in Drosophila melanogaster.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {18}, pages = {6773-6776}, pmid = {22798369}, issn = {1098-5336}, mesh = {Animals ; Drosophila melanogaster/immunology/*microbiology/*physiology ; *Metabolic Networks and Pathways ; RNA Viruses/immunology ; RNA, Small Interfering/*genetics/*metabolism ; Survival Analysis ; Wolbachia/*genetics/*metabolism ; }, abstract = {Wolbachia pipientis delays RNA virus-induced mortality in Drosophila spp. We investigated whether Wolbachia-mediated protection was dependent on the small interfering RNA (siRNA) pathway, a key antiviral defense. Compared to Wolbachia-free flies, virus-induced mortality was delayed in Wolbachia-infected flies with loss-of-function of siRNA pathway components, indicating that Wolbachia-mediated protection functions in the absence of the canonical siRNA pathway.}, } @article {pmid22763636, year = {2012}, author = {Simón, F and Siles-Lucas, M and Morchón, R and González-Miguel, J and Mellado, I and Carretón, E and Montoya-Alonso, JA}, title = {Human and animal dirofilariasis: the emergence of a zoonotic mosaic.}, journal = {Clinical microbiology reviews}, volume = {25}, number = {3}, pages = {507-544}, pmid = {22763636}, issn = {1098-6618}, mesh = {Animals ; Cats ; Communicable Diseases, Emerging/epidemiology/parasitology ; Dirofilaria immitis/immunology/metabolism/*pathogenicity ; Dirofilaria repens/immunology/metabolism/*pathogenicity ; Dirofilariasis/epidemiology/immunology/*parasitology/pathology ; Dogs ; Energy Metabolism ; Geography ; Helminth Proteins/immunology/metabolism ; Host-Parasite Interactions ; Humans ; Insect Vectors/parasitology ; Life Cycle Stages ; Population Dynamics ; Wolbachia/growth & development ; Zoonoses/epidemiology/*parasitology ; }, abstract = {Dirofilariasis represents a zoonotic mosaic, which includes two main filarial species (Dirofilaria immitis and D. repens) that have adapted to canine, feline, and human hosts with distinct biological and clinical implications. At the same time, both D. immitis and D. repens are themselves hosts to symbiotic bacteria of the genus Wolbachia, the study of which has resulted in a profound shift in the understanding of filarial biology, the mechanisms of the pathologies that they produce in their hosts, and issues related to dirofilariasis treatment. Moreover, because dirofilariasis is a vector-borne transmitted disease, their distribution and infection rates have undergone significant modifications influenced by global climate change. Despite advances in our knowledge of D. immitis and D. repens and the pathologies that they inflict on different hosts, there are still many unknown aspects of dirofilariasis. This review is focused on human and animal dirofilariasis, including the basic morphology, biology, protein composition, and metabolism of Dirofilaria species; the climate and human behavioral factors that influence distribution dynamics; the disease pathology; the host-parasite relationship; the mechanisms involved in parasite survival; the immune response and pathogenesis; and the clinical management of human and animal infections.}, } @article {pmid22738314, year = {2012}, author = {Toews, DP and Brelsford, A}, title = {The biogeography of mitochondrial and nuclear discordance in animals.}, journal = {Molecular ecology}, volume = {21}, number = {16}, pages = {3907-3930}, doi = {10.1111/j.1365-294X.2012.05664.x}, pmid = {22738314}, issn = {1365-294X}, mesh = {Amphibians/genetics ; Animals ; Birds/genetics ; Cell Nucleus/*genetics ; DNA, Mitochondrial/*genetics ; Fishes/genetics ; *Genetics, Population ; Haplotypes ; Insecta/genetics ; Mammals/genetics ; *Phylogeography ; Reptiles/genetics ; }, abstract = {Combining nuclear (nuDNA) and mitochondrial DNA (mtDNA) markers has improved the power of molecular data to test phylogenetic and phylogeographic hypotheses and has highlighted the limitations of studies using only mtDNA markers. In fact, in the past decade, many conflicting geographic patterns between mitochondrial and nuclear genetic markers have been identified (i.e. mito-nuclear discordance). Our goals in this synthesis are to: (i) review known cases of mito-nuclear discordance in animal systems, (ii) to summarize the biogeographic patterns in each instance and (iii) to identify common drivers of discordance in various groups. In total, we identified 126 cases in animal systems with strong evidence of discordance between the biogeographic patterns obtained from mitochondrial DNA and those observed in the nuclear genome. In most cases, these patterns are attributed to adaptive introgression of mtDNA, demographic disparities and sex-biased asymmetries, with some studies also implicating hybrid zone movement, human introductions and Wolbachia infection in insects. We also discuss situations where divergent mtDNA clades seem to have arisen in the absence of geographic isolation. For those cases where foreign mtDNA haplotypes are found deep within the range of a second taxon, data suggest that those mtDNA haplotypes are more likely to be at a high frequency and are commonly driven by sex-biased asymmetries and/or adaptive introgression. In addition, we discuss the problems with inferring the processes causing discordance from biogeographic patterns that are common in many studies. In many cases, authors presented more than one explanation for discordant patterns in a given system, which indicates that likely more data are required. Ideally, to resolve this issue, we see important future work shifting focus from documenting the prevalence of mito-nuclear discordance towards testing hypotheses regarding the drivers of discordance. Indeed, there is great potential for certain cases of mitochondrial introgression to become important natural systems within which to test the effect of different mitochondrial genotypes on whole-animal phenotypes.}, } @article {pmid22732604, year = {2012}, author = {Chen, SJ and Lu, F and Cheng, JA and Jiang, MX and Way, MO}, title = {Identification and biological role of the endosymbionts Wolbachia in rice water weevil (Coleoptera: Curculionidae).}, journal = {Environmental entomology}, volume = {41}, number = {3}, pages = {469-477}, doi = {10.1603/EN11195}, pmid = {22732604}, issn = {1938-2936}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/genetics ; China ; Female ; Molecular Sequence Data ; Multilocus Sequence Typing ; *Pest Control, Biological ; Phylogeny ; Polymerase Chain Reaction ; Reproduction/drug effects ; Tetracycline/pharmacology ; Texas ; Weevils/drug effects/*microbiology/physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia spp. are obligate intracellular bacteria present in reproductive tissues of many arthropod species. Wolbachia infection status and roles in host reproduction were studied in the rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera, Curculionidae), an introduced species in China. We examined Wolbachia infection status in five populations in China where it reproduces parthenogenetically, and one native population in Southeast Texas, where it reproduces bisexually. All populations were infected by Wolbachia, and all specimens in each population were infected by Wolbachia of a single strain. Phylogenetic analyses based on multilocus sequence typing system indicated that Wolbachia in non-native L. oryzophilus weevils diverges evidently from those in native weevils. After treatments with tetracycline, parthenogenetic weevils reduced the fecundity significantly and eggs were not viable. Our results suggest that Wolbachia are necessary for oocyte production in L oryzophilus.}, } @article {pmid22731846, year = {2012}, author = {Graham, RI and Grzywacz, D and Mushobozi, WL and Wilson, K}, title = {Wolbachia in a major African crop pest increases susceptibility to viral disease rather than protects.}, journal = {Ecology letters}, volume = {15}, number = {9}, pages = {993-1000}, doi = {10.1111/j.1461-0248.2012.01820.x}, pmid = {22731846}, issn = {1461-0248}, support = {BB/F004311/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Africa ; Animals ; Biological Control Agents ; Disease Susceptibility/veterinary ; Host-Parasite Interactions ; Insect Control ; Nucleopolyhedroviruses/*pathogenicity ; Population Dynamics ; Spodoptera/*microbiology/*virology ; Wolbachia ; }, abstract = {Wolbachia are common vertically transmitted endosymbiotic bacteria found in < 70% of insect species. They have generated considerable recent interest due to the capacity of some strains to protect their insect hosts against viruses and the potential for this to reduce vector competence of a range of human diseases, including dengue. In contrast, here we provide data from field populations of a major crop pest, African armyworm (Spodoptera exempta), which show that the prevalence and intensity of infection with a nucleopolydrovirus (SpexNPV) is positively associated with infection with three strains of Wolbachia. We also use laboratory bioassays to demonstrate that infection with one of these strains, a male-killer, increases host mortality due to SpexNPV by 6-14 times. These findings suggest that rather than protecting their lepidopteran host from viral infection, Wolbachia instead make them more susceptible. This finding potentially has implications for the biological control of other insect crop pests.}, } @article {pmid22727144, year = {2012}, author = {Sjödin, A and Svensson, K and Ohrman, C and Ahlinder, J and Lindgren, P and Duodu, S and Johansson, A and Colquhoun, DJ and Larsson, P and Forsman, M}, title = {Genome characterisation of the genus Francisella reveals insight into similar evolutionary paths in pathogens of mammals and fish.}, journal = {BMC genomics}, volume = {13}, number = {}, pages = {268}, pmid = {22727144}, issn = {1471-2164}, mesh = {Animals ; *Biological Evolution ; DNA Transposable Elements ; DNA, Bacterial/genetics ; Fishes/microbiology ; Francisella/*classification/genetics ; *Genome, Bacterial ; Mammals/microbiology ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Recombination, Genetic ; Sequence Analysis, DNA ; }, abstract = {BACKGROUND: Prior to this study, relatively few strains of Francisella had been genome-sequenced. Previously published Francisella genome sequences were largely restricted to the zoonotic agent F. tularensis. Only limited data were available for other members of the Francisella genus, including F. philomiragia, an opportunistic pathogen of humans, F. noatunensis, a serious pathogen of farmed fish, and other less well described endosymbiotic species.

RESULTS: We determined the phylogenetic relationships of all known Francisella species, including some for which the phylogenetic positions were previously uncertain. The genus Francisella could be divided into two main genetic clades: one included F. tularensis, F. novicida, F. hispaniensis and Wolbachia persica, and another included F. philomiragia and F. noatunensis.Some Francisella species were found to have significant recombination frequencies. However, the fish pathogen F. noatunensis subsp. noatunensis was an exception due to it exhibiting a highly clonal population structure similar to the human pathogen F. tularensis.

CONCLUSIONS: The genus Francisella can be divided into two main genetic clades occupying both terrestrial and marine habitats. However, our analyses suggest that the ancestral Francisella species originated in a marine habitat. The observed genome to genome variation in gene content and IS elements of different species supports the view that similar evolutionary paths of host adaptation developed independently in F. tularensis (infecting mammals) and F. noatunensis subsp. noatunensis (infecting fish).}, } @article {pmid22698088, year = {2013}, author = {Skaljac, M and Zanić, K and Hrnčić, S and Radonjić, S and Perović, T and Ghanim, M}, title = {Diversity and localization of bacterial symbionts in three whitefly species (Hemiptera: Aleyrodidae) from the east coast of the Adriatic Sea.}, journal = {Bulletin of entomological research}, volume = {103}, number = {1}, pages = {48-59}, doi = {10.1017/S0007485312000399}, pmid = {22698088}, issn = {1475-2670}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; *Bacterial Physiological Phenomena ; Croatia ; Female ; Hemiptera/genetics/*microbiology/*physiology ; In Situ Hybridization, Fluorescence ; Montenegro ; Polymerase Chain Reaction ; Species Specificity ; *Symbiosis ; }, abstract = {Several whitefly species (Hemiptera: Aleyrodidae) are cosmopolitan phloem-feeders that cause serious damage in numerous agricultural crops. All whitefly species harbor a primary bacterial symbiont and a diverse array of secondary symbionts which may influence several aspects of the insect's biology. We surveyed infections by secondary symbionts in Bemisia tabaci (Gennadius), Trialeurodes vaporariorum (Westwood) and Siphoninus phillyreae (Haliday) from areas in the east cost of the Adriatic Sea. Both the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) B. tabaci genetic groups were detected in Montenegro, whereas only the MED was confirmed in Croatia. Trialeurodes vaporariorum and S. phillyreae were found in all areas surveyed. MEAM1 and MED exhibited similarity to previously reported infections, while populations of T. vaporariorum from Montenegro harbored Rickettsia, Wolbachia and Cardinium in addition to previously reported Hamiltonella and Arsenopnohus. Siphoninus phillyreae harbored Hamiltonella, Wolbachia, Cardinium and Arsenophonus, with the latter appearing in two alleles. Multiple infections of all symbionts were common in the three insect species tested, with some reaching near fixation. Florescent in situ hybridization showed new localization patterns for Hamiltonella in S. phillyreae, and the morphology of the bacteriosome differed from that observed in other whitefly species. Our results show new infections with bacterial symbionts in the whitefly species studied. Infections with the same symbionts in reproductively isolated whitefly species confirm complex relationships between whiteflies and bacterial symbionts, and suggest possible horizontal transfer of some of these bacteria.}, } @article {pmid22685581, year = {2012}, author = {Zug, R and Hammerstein, P}, title = {Still a host of hosts for Wolbachia: analysis of recent data suggests that 40% of terrestrial arthropod species are infected.}, journal = {PloS one}, volume = {7}, number = {6}, pages = {e38544}, pmid = {22685581}, issn = {1932-6203}, mesh = {Algorithms ; Animals ; Arthropods/classification/*microbiology ; *Host Specificity ; Models, Statistical ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular bacteria that manipulate the reproduction of their arthropod hosts in remarkable ways. They are predominantly transmitted vertically from mother to offspring but also occasionally horizontally between species. In doing so, they infect a huge range of arthropod species worldwide. Recently, a statistical analysis estimated the infection frequency of Wolbachia among arthropod hosts to be 66%. At the same time, the authors of this analysis highlighted some weaknesses of the underlying data and concluded that in order to improve the estimate, a larger number of individuals per species should be assayed and species be chosen more randomly. Here we apply the statistical approach to a more appropriate data set from a recent survey that tested both a broad range of species and a sufficient number of individuals per species. Indeed, we find a substantially different infection frequency: We now estimate the proportion of Wolbachia-infected species to be around 40% which is lower than the previous estimate but still points to a surprisingly high number of arthropods harboring the bacteria. Notwithstanding this difference, we confirm the previous result that, within a given species, typically most or only a few individuals are infected. Moreover, we extend our analysis to include several reproductive parasites other than Wolbachia that were also screened for in the aforementioned empirical survey. For these symbionts we find a large variation in estimated infection frequencies and corroborate the finding that Wolbachia are the most abundant endosymbionts among arthropod species.}, } @article {pmid22685158, year = {2012}, author = {Gueguen, G and Onemola, B and Govind, S}, title = {Association of a new Wolbachia strain with, and its effects on, Leptopilina victoriae, a virulent wasp parasitic to Drosophila spp.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {16}, pages = {5962-5966}, pmid = {22685158}, issn = {1098-5336}, support = {G12 MD007603/MD/NIMHD NIH HHS/United States ; G12 RR003060/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; Drosophila/parasitology ; Host Specificity ; Molecular Sequence Data ; Oocytes/microbiology ; Phylogeny ; Sequence Analysis, DNA ; Wasps/*microbiology/physiology ; Wolbachia/*classification/genetics/*isolation & purification/physiology ; }, abstract = {Wolbachia bacteria are ubiquitous intracellular bacteria of arthropods. Often considered reproductive parasites, they can benefit certain host species. We describe a new Wolbachia strain from Leptopilina victoriae, a Drosophila wasp. The strain is closely related to Wolbachia from Culex sp. Located to the posterior poles of oocytes, it manipulates its host's reproduction by inducing a male development type of cytoplasmic incompatibility. We also report its diverse effects on the wasp's life history traits.}, } @article {pmid22679870, year = {2012}, author = {Hurst, TP and Pittman, G and O'Neill, SL and Ryan, PA and Nguyen, HL and Kay, BH}, title = {Impacts of Wolbachia infection on predator prey relationships: evaluating survival and horizontal transfer between wMelPop infected Aedes aegypti and its predators.}, journal = {Journal of medical entomology}, volume = {49}, number = {3}, pages = {624-630}, doi = {10.1603/me11277}, pmid = {22679870}, issn = {0022-2585}, mesh = {Aedes/*microbiology ; Animals ; Copepoda ; Larva/microbiology ; Poecilia ; *Predatory Behavior ; Spiders ; Wolbachia/*physiology ; }, abstract = {The wMelPop strain of Wolbachia is currently being investigated for its potential use as a biological control agent to reduce the ability of Aedes aegypti (L.) mosquitoes to transmit dengue viruses. The survival of a potential wMelPop infected Ae. aegypti strain for field release is important as a higher susceptibility to predation in the wMelPop strain could result in difficulties in achieving fixation. We investigated immature and adult survival as a function of susceptibility to predation by six naturally occurring predator species; cyclopoid copepods, fish, predatory Toxorhynchites mosquito larvae and a salticid jumping spider. The trials indicated that wMelPop infected and uninfected Ae. aegypti larvae and adults were equally susceptible to predation to all six tested predators. In addition to evaluating any potential fitness costs to the infected host, we were unable to demonstrate horizontal transfer of wMelPop via consumption of infected Ae. aegypti larvae to the above predators. That susceptibility to predation was consistent across mosquito life stage, predator species and experimental venue is strong evidence that despite the neurotrophic and extensive nature of wMelPop infection, behavioral changes are not occurring, or at least not a determining factor in survival when exposed to a predator. Based on our results and the ecology of Wolbachia and mosquito predators, horizontal transfer of wMelPop from Ae. aegypti into naturally occurring predators is not cause for concern.}, } @article {pmid22679853, year = {2012}, author = {Morningstar, RJ and Hamer, GL and Goldberg, TL and Huang, S and Andreadis, TG and Walker, ED}, title = {Diversity of Wolbachia pipientis strain wPip in a genetically admixtured, above-ground Culex pipiens (Diptera: Culicidae) population: association with form molestus ancestry and host selection patterns.}, journal = {Journal of medical entomology}, volume = {49}, number = {3}, pages = {474-481}, pmid = {22679853}, issn = {0022-2585}, support = {R37 AI021884/AI/NIAID NIH HHS/United States ; R37AI21884/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/genetics ; Culex/*microbiology ; Female ; Genetic Variation ; Host Specificity ; Wolbachia/*genetics/virology ; }, abstract = {Analysis of molecular genetic diversity in nine marker regions of five genes within the bacteriophage WO genomic region revealed high diversity of the Wolbachia pipentis strain wPip in a population of Culex pipiens L. sampled in metropolitan Chicago, IL. From 166 blood fed females, 50 distinct genetic profiles of wPip were identified. Rarefaction analysis suggested a maximum of 110 profiles out of a possible 512 predicted by combinations of the nine markers. A rank-abundance curve showed that few strains were common and most were rare. Multiple regression showed that markers associated with gene Gp2d, encoding a partial putative capsid protein, were significantly associated with ancestry of individuals either to form molestus or form pipiens, as determined by prior microsatellite allele frequency analysis. None of the other eight markers was associated with ancestry to either form, nor to ancestry to Cx. quinquefasciatus Say. Logistic regression of host choice (mammal vs. avian) as determined by bloodmeal analysis revealed that significantly fewer individuals that had fed on mammals had the Gp9a genetic marker (58.5%) compared with avian-fed individuals (88.1%). These data suggest that certain wPip molecular genetic types are associated with genetic admixturing in the Cx. pipiens complex of metropolitan Chicago, IL, and that the association extends to phenotypic variation related to host preference.}, } @article {pmid22677398, year = {2012}, author = {Yamauchi, A and Telschow, A}, title = {Bistability of endosymbiont evolution of genome size and host sex control.}, journal = {Journal of theoretical biology}, volume = {309}, number = {}, pages = {58-66}, doi = {10.1016/j.jtbi.2012.05.014}, pmid = {22677398}, issn = {1095-8541}, mesh = {Animals ; Computer Simulation ; *Evolution, Molecular ; Female ; Genome Size/*genetics ; Male ; Models, Biological ; *Sex Ratio ; Symbiosis/*genetics ; }, abstract = {Eukaryotic organisms often harbor several genetic factors in their cytoplasm. These cytoplasmic genetic elements (CGEs) include both eukaryotic organelles (mitochondria, chloroplasts) and bacterial endosymbionts, which have evolved from free-living bacteria. A common feature of CGEs is their cytoplasmic inheritance from mother to offspring. A striking difference is that some CGEs have evolved a short genome size (e.g., animal mitochondria), while others cause a sex ratio distortion (SRD) in their hosts (e.g., Wolbachia). In this study, we sought to resolve the evolution of these endosymbiont properties using a population genetics approach. Our model divides the endosymbiont genome into a functional part and a part that can cause SRD, and our results indicate that the cytoplasmic inheritance system at the initiation of symbiosis plays a key role in determining the evolutionary trajectory of CGEs. We show that in endosymbiotic evolution, two states can be bistable, depending on the parameters. The evolution of the cytoplasmic inheritance system from biparental to uniparental can result in hysteresis in the evolution of cytoplasmic symbionts.}, } @article {pmid22676974, year = {2012}, author = {Rugman-Jones, PF and Hoddle, MS and Amrich, R and Heraty, JM and Stouthamer-Ingel, CE and Stouthamer, R}, title = {Phylogeographic structure, outbreeding depression, and reluctant virgin oviposition in the bean thrips, Caliothrips fasciatus (Pergande) (Thysanoptera: Thripidae), in California.}, journal = {Bulletin of entomological research}, volume = {102}, number = {6}, pages = {698-709}, doi = {10.1017/S0007485312000302}, pmid = {22676974}, issn = {1475-2670}, mesh = {Animals ; California ; Crosses, Genetic ; Electron Transport Complex IV/genetics ; Female ; Genetic Variation ; Male ; Oviposition ; Parthenogenesis ; Phylogeography ; *Reproductive Isolation ; Symbiosis ; Thysanoptera/*genetics/microbiology ; Wolbachia/physiology ; }, abstract = {Bean thrips, Caliothrips fasciatus, is native to western North America. Once considered a pest of several crops in its native area, its pest status has waned over recent decades. However, due to its habit of aggregating in the navel of navel oranges, bean thrips remains economically important because some countries importing oranges from California have designated it a quarantine pest. Despite continued propagule pressure, bean thrips has never established outside North America. We examined genetic variation in mitochondrial DNA among Californian populations of C. fasciatus and found that potentially two cryptic species are present (supported by Kimura 2-P distances): a common widespread form B and a rarer form A with a very limited distribution. Form B showed strong phylogeographic structure, with many haplotypes having a limited geographic distribution. Inter-population crossing experiments between three geographically isolated populations of form B resulted in the production of some female offspring, indicating a degree of compatibility between these populations of this haplodiploid species. However, substantial outbreeding depression was also detected. A low frequency of offspring production by hetero-population pairs was evidence of pre-mating isolation, while post-mating isolation was also evident in the elevated mortality of fertilized eggs in successful hetero-population crosses. One surprising finding was the total lack of offspring production by virgin females when isolated individually. However, virgin females did produce sons in the presence of other virgin females. A test for the presence of Wolbachia showed that form B was not infected, but that some populations of the rarer form A were.}, } @article {pmid22675165, year = {2012}, author = {Hancock, PA and Godfray, HC}, title = {Modelling the spread of Wolbachia in spatially heterogeneous environments.}, journal = {Journal of the Royal Society, Interface}, volume = {9}, number = {76}, pages = {3045-3054}, pmid = {22675165}, issn = {1742-5662}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Age Factors ; Animals ; *Environment ; Host-Pathogen Interactions ; Insecta/*microbiology ; *Models, Biological ; Population Density ; Population Dynamics ; *Symbiosis ; Wolbachia/*growth & development ; }, abstract = {The endosymbiont Wolbachia infects a large number of insect species and is capable of rapid spread when introduced into a novel host population. The bacteria spread by manipulating their hosts' reproduction, and their dynamics are influenced by the demographic structure of the host population and patterns of contact between individuals. Reaction-diffusion models of the spatial spread of Wolbachia provide a simple analytical description of their spatial dynamics but do not account for significant details of host population dynamics. We develop a metapopulation model describing the spatial dynamics of Wolbachia in an age-structured host insect population regulated by juvenile density-dependent competition. The model produces similar dynamics to the reaction-diffusion model in the limiting case where the host's habitat quality is spatially homogeneous and Wolbachia has a small effect on host fitness. When habitat quality varies spatially, Wolbachia spread is usually much slower, and the conditions necessary for local invasion are strongly affected by immigration of insects from surrounding regions. Spread is most difficult when variation in habitat quality is spatially correlated. The results show that spatial variation in the density-dependent competition experienced by juvenile host insects can strongly affect the spread of Wolbachia infections, which is important to the use of Wolbachia to control insect vectors of human disease and other pests.}, } @article {pmid22674991, year = {2012}, author = {Shaw, AE and Veronesi, E and Maurin, G and Ftaich, N and Guiguen, F and Rixon, F and Ratinier, M and Mertens, P and Carpenter, S and Palmarini, M and Terzian, C and Arnaud, F}, title = {Drosophila melanogaster as a model organism for bluetongue virus replication and tropism.}, journal = {Journal of virology}, volume = {86}, number = {17}, pages = {9015-9024}, pmid = {22674991}, issn = {1098-5514}, support = {BBS/E/I/00001701/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; G0801822/MRC_/Medical Research Council/United Kingdom ; MC_U130115834/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/6/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Bluetongue/*virology ; Bluetongue virus/genetics/*physiology ; Cattle ; Cattle Diseases/*virology ; Cell Line ; Ceratopogonidae/virology ; *Disease Models, Animal ; Drosophila melanogaster/genetics/*virology ; Insect Vectors/virology ; *Viral Tropism ; *Virus Replication ; }, abstract = {Bluetongue virus (BTV) is the etiological agent of bluetongue (BT), a hemorrhagic disease of ruminants that can cause high levels of morbidity and mortality. BTV is an arbovirus transmitted between its ruminant hosts by Culicoides biting midges (Diptera: Ceratopogonidae). Recently, Europe has experienced some of the largest BT outbreaks ever recorded, including areas with no known history of the disease, leading to unprecedented economic and animal welfare issues. The current lack of genomic resources and genetic tools for Culicoides restricts any detailed study of the mechanisms involved in the virus-insect interactions. In contrast, the genome of the fruit fly (Drosophila melanogaster) has been successfully sequenced, and it is used extensively as a model of molecular pathways due to the existence of powerful genetic technology. In this study, D. melanogaster is investigated as a model for the replication and tropism of BTV. Using reverse genetics, a modified BTV-1 that expresses the fluorescent mCherry protein fused to the viral nonstructural protein NS3 (BTV-1/NS3mCherry) was generated. We demonstrate that BTV-1/NS3mCherry is not only replication competent as it retains many characteristics of the wild-type virus but also replicates efficiently in D. melanogaster after removal of the bacterial endosymbiont Wolbachia pipientis by antibiotic treatment. Furthermore, confocal microscopy shows that the tissue tropism of BTV-1/NS3mCherry in D. melanogaster resembles that described previously for BTV in Culicoides. Overall, the data presented in this study demonstrate the feasibility of using D. melanogaster as a genetic model to investigate BTV-insect interactions that cannot be otherwise addressed in vector species.}, } @article {pmid22671555, year = {2012}, author = {Xiao, JH and Wang, NX and Murphy, RW and Cook, J and Jia, LY and Huang, DW}, title = {Wolbachia infection and dramatic intraspecific mitochondrial DNA divergence in a fig wasp.}, journal = {Evolution; international journal of organic evolution}, volume = {66}, number = {6}, pages = {1907-1916}, doi = {10.1111/j.1558-5646.2011.01561.x}, pmid = {22671555}, issn = {1558-5646}, mesh = {Animals ; Base Sequence ; DNA Primers ; DNA, Mitochondrial/genetics ; Gene Transfer, Horizontal ; Gram-Negative Bacterial Infections/*genetics/microbiology ; Haplotypes ; Polymerase Chain Reaction ; Wasps/*genetics ; Wolbachia/*isolation & purification ; }, abstract = {Mitochondria and Wolbachia are maternally inherited genomes that exhibit strong linkage disequilibrium in many organisms. We surveyed Wolbachia infections in 187 specimens of the fig wasp species, Ceratosolen solmsi, and found an infection prevalence of 89.3%. DNA sequencing of 20 individuals each from Wolbachia-infected and -uninfected subpopulations revealed extreme mtDNA divergence (up to 9.2% and 15.3% in CO1 and cytochrome b, respectively) between infected and uninfected wasps. Further, mtDNA diversity was significantly reduced within the infected group. Our sequencing of a large part of the mitochondrial genome from both Wolbachia-infected and -uninfected individuals revealed that high sequence divergence is common throughout the mitochondrial genome. These patterns suggest a partial selective sweep of mitochondria subsequent to the introduction of Wolbachia into C. solsmi, by hybrid introgression from a related species.}, } @article {pmid22669278, year = {2012}, author = {Lu, MH and Zhang, KJ and Hong, XY}, title = {Tripartite associations among bacteriophage WO, Wolbachia, and host affected by temperature and age in Tetranychus urticae.}, journal = {Experimental & applied acarology}, volume = {58}, number = {3}, pages = {207-220}, pmid = {22669278}, issn = {1572-9702}, mesh = {Animals ; Bacteriophages/genetics/*physiology ; Cloning, Molecular ; Phylogeny ; *Temperature ; Tetranychidae/genetics/*microbiology ; Time Factors ; Wolbachia/genetics/physiology/*virology ; }, abstract = {A phage density model of cytoplasmic incompatibility (CI), which means lytic phages reduce bacterial density associated with CI, significantly enhances our understanding of the tripartite associations among bacteriophage WO, Wolbachia and host. However, WO may alternate between lytic and lysogenic life cycles or change phage production under certain conditions including temperature, host age and host species background. Here, extreme temperatures can induce an alteration in the life cycle of WO and change the tripartite associations among WO, Wolbachia and CI. Based on the accumulation of the WO load, WO can transform into the lytic life cycle with increasing age. These findings confirmed that the environment plays an important role in the associations among WO, Wolbachia and host.}, } @article {pmid24672682, year = {2012}, author = {Gentil, K and Hoerauf, A and Pearlman, E}, title = {Differential induction of Th2- and Th1-associated responses by filarial antigens and endosymbiotic Wolbachia in a murine model of river blindness.}, journal = {European journal of microbiology & immunology}, volume = {2}, number = {2}, pages = {134-139}, pmid = {24672682}, issn = {2062-509X}, abstract = {Immune responses to filarial parasites like the river blindness inducing Onchocerca volvulus are obscured by combined reactions to the filarial nematodes themselves and their endosymbiont bacteria Wolbachia. Overall, infection with filarial nematodes induces a strong Th2 response characterized by IL-5 production and to a lesser degree a Th1 response and IFNγ production. Neutrophil and eosinophil infiltration into the corneal stroma are hallmark features of Onchocerca volvulus stimulation in a mouse model of river blindness. To determine the splenic and corneal response to filarial antigens in the absence of Wolbachia, C57BL/6 mice were immunized subcutaneously with either endosymbiotic Wolbachia alone, a soluble extract from the filaria Acanthocheilonema viteae that does not contain Wolbachia, or both, and injected into the corneal stroma. Neutrophil and eosinophil infiltration into the cornea was assessed by immunohistochemistry. In addition, Th1- and Th2-associated responses to filaria or Wolbachia were investigated by determining IL-5 and IFN-γ production by splenocytes. We found that A. viteae in the absence of Wolbachia induced IL-5 production and eosinophil infiltration, but not IFN-γ. Conversely, Wolbachia induced IFN-γ production and no migration of eosinophils. There was no difference in neutrophil infiltration. Together, these findings demonstrate a distinct Th-associated phenotype induced by filaria and Wolbachia.}, } @article {pmid22649020, year = {2012}, author = {Turba, ME and Zambon, E and Zannoni, A and Russo, S and Gentilini, F}, title = {Detection of Wolbachia DNA in blood for diagnosing filaria-associated syndromes in cats.}, journal = {Journal of clinical microbiology}, volume = {50}, number = {8}, pages = {2624-2630}, pmid = {22649020}, issn = {1098-660X}, mesh = {Animals ; Bacterial Proteins/genetics ; Blood/*microbiology ; Cat Diseases/*diagnosis ; Cats ; DNA, Bacterial/genetics/*isolation & purification ; Dirofilaria immitis/*microbiology ; Filariasis/diagnosis/*veterinary ; Polymerase Chain Reaction/*methods ; Sensitivity and Specificity ; Veterinary Medicine/methods ; Wolbachia/genetics/*isolation & purification ; }, abstract = {A fundamental role for the endosymbiotic bacteria Wolbachia pipientis in the pathogenesis of Dirofilaria immitis infections has emerged in recent years. Diagnostic opportunities arising from this breakthrough have not yet been fully exploited. This study was aimed at developing conventional and real-time PCR assays to carry out a molecular survey in a convenience sample of cats living in an area where D. immitis is endemic and to evaluate the detection of bacterial DNA in blood as a surrogate assay for diagnosing filaria-associated syndromes in cats. COI and FtsZ loci were used as targets for D. immitis and Wolbachia PCR assays, respectively, and real-time TaqMan PCR assays were used only for Wolbachia. A convenience sample of 307 disease-affected or healthy cats examined at a University facility were PCR tested, and their medical records were investigated. Conventional nested PCR for Wolbachia amplified the endosymbionts of both D. immitis and D. repens, while real-time PCR was highly specific only for the former. Observed prevalences of 0.3 and 10.4% were found using conventional nested PCR assays for D. immitis and real-time PCR for Wolbachia, respectively. Similar prevalences were established using the Wolbachia nested PCR (98% concordance with real-time PCR). The group of Wolbachia-positive samples had a significantly higher proportion of subjects with respiratory signs (29.0% versus 9.7%; P = 0.002). The findings of this study indicate that a highly sensitive PCR assay can be used to detect the Wolbachia organism in the peripheral blood of cats with respiratory signs.}, } @article {pmid22645363, year = {2012}, author = {Voronin, D and Cook, DA and Steven, A and Taylor, MJ}, title = {Autophagy regulates Wolbachia populations across diverse symbiotic associations.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {109}, number = {25}, pages = {E1638-46}, pmid = {22645363}, issn = {1091-6490}, mesh = {Animals ; *Autophagy ; Bacterial Proteins/metabolism ; Brugia malayi/metabolism/microbiology ; Cell Line/metabolism ; Drosophila melanogaster/metabolism ; Nematoda/microbiology ; Subcellular Fractions/metabolism ; *Symbiosis ; TOR Serine-Threonine Kinases/antagonists & inhibitors/metabolism ; Wolbachia/*physiology ; }, abstract = {Wolbachia are widespread and abundant intracellular symbionts of arthropods and filarial nematodes. Their symbiotic relationships encompass obligate mutualism, commensalism, parasitism, and pathogenicity. A consequence of these diverse associations is that Wolbachia encounter a wide range of host cells and intracellular immune defense mechanisms of invertebrates, which they must evade to maintain their populations and spread to new hosts. Here we show that autophagy, a conserved intracellular defense mechanism and regulator of cell homeostasis, is a major immune recognition and regulatory process that determines the size of Wolbachia populations. The regulation of Wolbachia populations by autophagy occurs across all distinct symbiotic relationships and can be manipulated either chemically or genetically to modulate the Wolbachia population load. The recognition and activation of host autophagy is particularly apparent in rapidly replicating strains of Wolbachia found in somatic tissues of Drosophila and filarial nematodes. In filarial nematodes, which host a mutualistic association with Wolbachia, the use of antibiotics such as doxycycline to eliminate Wolbachia has emerged as a promising approach to their treatment and control. Here we show that the activation of host nematode autophagy reduces bacterial loads to the same magnitude as antibiotic therapy; thus we identify a bactericidal mode of action targeting Wolbachia that can be exploited for the development of chemotherapeutic agents against onchocerciasis, lymphatic filariasis, and heartworm.}, } @article {pmid22634096, year = {2012}, author = {Wiwatanaratanabutr, I}, title = {WITHDRAWN: Geographic distribution of Wolbachia infection in mosquitoes from Thailand.}, journal = {Journal of invertebrate pathology}, volume = {}, number = {}, pages = {}, doi = {10.1016/j.jip.2012.04.010}, pmid = {22634096}, issn = {1096-0805}, abstract = {This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.}, } @article {pmid22620700, year = {2012}, author = {Crowder, CD and Matthews, HE and Rounds, MA and Li, F and Schutzer, SE and Sampath, R and Hofstadler, SA and Ecker, DJ and Eshoo, MW}, title = {Detection of heartworm infection in dogs via PCR amplification and electrospray ionization mass spectrometry of nucleic acid extracts from whole blood samples.}, journal = {American journal of veterinary research}, volume = {73}, number = {6}, pages = {854-859}, doi = {10.2460/ajvr.73.6.854}, pmid = {22620700}, issn = {1943-5681}, support = {2R44AI077156-02/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Base Sequence ; DNA Primers/genetics ; DNA, Ribosomal/genetics ; Dirofilariasis/*diagnosis/genetics ; Dog Diseases/*diagnosis/genetics ; Dogs ; Mass Spectrometry/methods/veterinary ; Molecular Sequence Data ; Nucleic Acids/*blood ; Polymerase Chain Reaction/methods/veterinary ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Analysis, DNA/veterinary ; }, abstract = {OBJECTIVE: To develop and evaluate a rapid and accurate assay involving PCR amplification and electrospray ionization mass spectrometry of nucleic acid extracts from whole blood samples for the detection of Dirofilaria immitis infection in dogs.

SAMPLE: Whole blood nucleic acid extracts from 29 dogs experimentally infected with D immitis (and in which circulating D immitis antigen was detected) and 10 uninfected dogs.

PROCEDURES: 16 of the 29 whole blood samples from infected dogs were examined at the time of collection for circulating microfilaria. Nucleic acids were extracted from all whole blood specimens and underwent PCR amplification with 12 PCR primer pairs designed to detect a wide range of pathogens (including the Wolbachia endosymbiont of D immitis) and electrospray ionization mass spectrometry.

RESULTS: On the basis of assay results, heartworm infection was detected in 13 of 13 antigen-positive dogs of unknown microfilaria status, 11 of 11 antigen-positive dogs with circulating microfilaria, 0 of 3 antigen-positive dogs tested at 3 months after larval infection, 0 of 2 antigen-positive dogs with occult infections, and 0 of 10 uninfected dogs.

With the assay under investigation, it was possible to identify D immitis infection in dogs with circulating microfilaria via detection of the obligate Wolbachia endosymbiont of D immitis. It was not possible to identify dogs with occult infections, which suggested that circulating microfilaria must be present to detect infection with this assay, although further studies would be required to verify that finding.}, } @article {pmid22609369, year = {2012}, author = {Metcalf, JA and Bordenstein, SR}, title = {The complexity of virus systems: the case of endosymbionts.}, journal = {Current opinion in microbiology}, volume = {15}, number = {4}, pages = {546-552}, pmid = {22609369}, issn = {1879-0364}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; T32 GM007347/GM/NIGMS NIH HHS/United States ; T32 GM07347/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/microbiology ; Bacteriophages/*genetics/pathogenicity/physiology ; DNA, Viral/*genetics ; Evolution, Molecular ; Host-Pathogen Interactions ; *Symbiosis/genetics ; Wolbachia/*physiology/*virology ; }, abstract = {Host-microbe symbioses involving bacterial endosymbionts comprise some of the most intimate and long-lasting interactions on the planet. While restricted gene flow might be expected due to their intracellular lifestyle, many endosymbionts, especially those that switch hosts, are rampant with mobile DNA and bacteriophages. One endosymbiont, Wolbachia pipientis, infects a vast number of arthropod and nematode species and often has a significant portion of its genome dedicated to prophage sequences of a virus called WO. This phage has challenged fundamental theories of bacteriophage and endosymbiont evolution, namely the phage Modular Theory and bacterial genome stability in obligate intracellular species. WO has also opened up exciting windows into the tripartite interactions between viruses, bacteria, and eukaryotes.}, } @article {pmid22586066, year = {2012}, author = {Schiefer, A and Schmitz, A and Schäberle, TF and Specht, S and Lämmer, C and Johnston, KL and Vassylyev, DG and König, GM and Hoerauf, A and Pfarr, K}, title = {Corallopyronin A specifically targets and depletes essential obligate Wolbachia endobacteria from filarial nematodes in vivo.}, journal = {The Journal of infectious diseases}, volume = {206}, number = {2}, pages = {249-257}, pmid = {22586066}, issn = {1537-6613}, support = {R01 GM074840/GM/NIGMS NIH HHS/United States ; GM074840/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/cytology/microbiology ; Animals ; Cell Line ; Contraindications ; DNA-Directed RNA Polymerases/antagonists & inhibitors/chemistry ; Drug Resistance, Bacterial ; Enzyme Inhibitors/chemistry/pharmacology ; Female ; Filariasis/drug therapy/parasitology ; Filaricides/pharmacology ; Filarioidea/*microbiology ; Lactones/chemistry/*pharmacology ; Mice ; Mice, Inbred BALB C ; Molecular Conformation ; Rifampin/pharmacology ; Symbiosis ; Wolbachia/*drug effects/enzymology ; }, abstract = {Doxycycline and rifampicin deplete essential Wolbachia from filarial nematodes that cause lymphatic filariasis or onchocerciasis, resulting in blocked worm development and death. However, doxycycline is contraindicated for children and pregnant/breastfeeding women, as is rifampicin in the latter group with the additional specter of possible resistance development in Mycobacterium spp. Novel antibiotics with a narrower spectrum would aid in eliminating filarial diseases. Corallococcus coralloides synthesizes corallopyronin A, a noncompetitive inhibitor of RNA polymerase ineffective against Mycobacterium spp. Corallopyronin A depleted Wolbachia from infected insect cells (1.89 Thus the antibiotic is effective against intracellular bacteria despite the many intervening surfaces (blood vessels, pleura, worm cuticle) and membranes (worm cell, vesicle, Wolbachia inner and outer membranes). Corallopyronin A is an antibiotic to develop further for filariasis elimination without concern for cross-resistance development in tuberculosis.}, } @article {pmid22583543, year = {2012}, author = {Kumar, N and Creasy, T and Sun, Y and Flowers, M and Tallon, LJ and Dunning Hotopp, JC}, title = {Efficient subtraction of insect rRNA prior to transcriptome analysis of Wolbachia-Drosophila lateral gene transfer.}, journal = {BMC research notes}, volume = {5}, number = {}, pages = {230}, pmid = {22583543}, issn = {1756-0500}, mesh = {Actins/genetics ; Animals ; Drosophila/*genetics/*microbiology ; Gene Expression Profiling/*methods ; Gene Transfer, Horizontal/*genetics ; Genes, Insect/genetics ; RNA, Messenger/genetics/metabolism ; RNA, Ribosomal, 18S/genetics/*isolation & purification ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Numerous methods exist for enriching bacterial or mammalian mRNA prior to transcriptome experiments. Yet there persists a need for methods to enrich for mRNA in non-mammalian animal systems. For example, insects contain many important and interesting obligate intracellular bacteria, including endosymbionts and vector-borne pathogens. Such obligate intracellular bacteria are difficult to study by traditional methods. Therefore, genomics has greatly increased our understanding of these bacteria. Efficient subtraction methods are needed for removing both bacteria and insect rRNA in these systems to enable transcriptome-based studies.

FINDINGS: A method is described that efficiently removes >95% of insect rRNA from total RNA samples, as determined by microfluidics and transcriptome sequencing. This subtraction yielded a 6.2-fold increase in mRNA abundance. Such a host rRNA-depletion strategy, in combination with bacterial rRNA depletion, is necessary to analyze transcription of obligate intracellular bacteria. Here, transcripts were identified that arise from a lateral gene transfer of an entire Wolbachia bacterial genome into a Drosophila ananassae chromosome. In this case, an rRNA depletion strategy is preferred over polyA-based enrichment since transcripts arising from bacteria-to-animal lateral gene transfer may not be poly-adenylated.

CONCLUSIONS: This enrichment method yields a significant increase in mRNA abundance when poly-A selection is not suitable. It can be used in combination with bacterial rRNA subtraction to enable experiments to simultaneously measure bacteria and insect mRNA in vector and endosymbiont biology experiments.}, } @article {pmid22567162, year = {2012}, author = {Smith, MA and Bertrand, C and Crosby, K and Eveleigh, ES and Fernandez-Triana, J and Fisher, BL and Gibbs, J and Hajibabaei, M and Hallwachs, W and Hind, K and Hrcek, J and Huang, DW and Janda, M and Janzen, DH and Li, Y and Miller, SE and Packer, L and Quicke, D and Ratnasingham, S and Rodriguez, J and Rougerie, R and Shaw, MR and Sheffield, C and Stahlhut, JK and Steinke, D and Whitfield, J and Wood, M and Zhou, X}, title = {Wolbachia and DNA barcoding insects: patterns, potential, and problems.}, journal = {PloS one}, volume = {7}, number = {5}, pages = {e36514}, pmid = {22567162}, issn = {1932-6203}, mesh = {Animals ; DNA Barcoding, Taxonomic/*methods ; DNA, Mitochondrial/genetics ; Insecta/classification/*genetics/*microbiology ; Phylogeny ; Wolbachia/*genetics ; }, abstract = {Wolbachia is a genus of bacterial endosymbionts that impacts the breeding systems of their hosts. Wolbachia can confuse the patterns of mitochondrial variation, including DNA barcodes, because it influences the pathways through which mitochondria are inherited. We examined the extent to which these endosymbionts are detected in routine DNA barcoding, assessed their impact upon the insect sequence divergence and identification accuracy, and considered the variation present in Wolbachia COI. Using both standard PCR assays (Wolbachia surface coding protein--wsp), and bacterial COI fragments we found evidence of Wolbachia in insect total genomic extracts created for DNA barcoding library construction. When >2 million insect COI trace files were examined on the Barcode of Life Datasystem (BOLD) Wolbachia COI was present in 0.16% of the cases. It is possible to generate Wolbachia COI using standard insect primers; however, that amplicon was never confused with the COI of the host. Wolbachia alleles recovered were predominantly Supergroup A and were broadly distributed geographically and phylogenetically. We conclude that the presence of the Wolbachia DNA in total genomic extracts made from insects is unlikely to compromise the accuracy of the DNA barcode library; in fact, the ability to query this DNA library (the database and the extracts) for endosymbionts is one of the ancillary benefits of such a large scale endeavor--which we provide several examples. It is our conclusion that regular assays for Wolbachia presence and type can, and should, be adopted by large scale insect barcoding initiatives. While COI is one of the five multi-locus sequence typing (MLST) genes used for categorizing Wolbachia, there is limited overlap with the eukaryotic DNA barcode region.}, } @article {pmid22562106, year = {2012}, author = {Ventura, IM and Martins, AB and Lyra, ML and Andrade, CA and Carvalho, KA and Klaczko, LB}, title = {Spiroplasma in Drosophila melanogaster populations: prevalence, male-killing, molecular identification, and no association with Wolbachia.}, journal = {Microbial ecology}, volume = {64}, number = {3}, pages = {794-801}, pmid = {22562106}, issn = {1432-184X}, mesh = {Animals ; Brazil ; DNA, Bacterial/genetics/isolation & purification ; Drosophila melanogaster/*microbiology ; Female ; Male ; Phylogeny ; Polymerase Chain Reaction ; Prevalence ; Sex Factors ; Sex Ratio ; Species Specificity ; Spiroplasma/classification/*genetics/isolation & purification/*physiology ; Symbiosis ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Spiroplasma endosymbionts are maternally transmitted bacteria that may kill infected sons resulting in the production of female-biased broods. The prevalence of male killers varies considerably both between and within species. Here, we evaluate the spatial and temporal status of male-killing and non-male-killing Spiroplasma infection in three Brazilian populations of Drosophila melanogaster, nearly a decade after the first occurrence report for this species. The incidence of the male-killing Spiroplasma ranged from close to 0 to 17.7 % (so far the highest estimate for a Drosophila species) with a suggestion of temporal decline in a population. We also found non-male-killing Spiroplasma coexisting in one population at lower prevalence (3-5 %), and we did not detect it in the other two. This may be taken as a suggestion of a spreading advantage conferred by the male-killing strategy. Sequencing two loci, we identified the phylogenetic position of Spiroplasma strains from the three localities, showing that all strains group closely in the poulsonii clade. Due to intensive sampling effort, we were able to test the association between Spiroplasma infections and another widespread endosymbiont, Wolbachia, whose prevalence ranged from 81.8 to 100 %. The prevalence of Wolbachia did not differ between Spiroplasma-infected and uninfected strains in our largest sample nor were the prevalences of the two endosymbionts associated across localities.}, } @article {pmid22558418, year = {2012}, author = {Hughes, GL and Pike, AD and Xue, P and Rasgon, JL}, title = {Invasion of Wolbachia into Anopheles and other insect germlines in an ex vivo organ culture system.}, journal = {PloS one}, volume = {7}, number = {4}, pages = {e36277}, pmid = {22558418}, issn = {1932-6203}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology ; Female ; Host-Pathogen Interactions ; Male ; Organ Culture Techniques/*methods ; Ovary/*microbiology ; Pest Control, Biological/*methods ; Species Specificity ; Testis/*microbiology ; Time Factors ; Wolbachia/*physiology ; }, abstract = {The common bacterial endosymbiont Wolbachia manipulates its host's reproduction to promote its own maternal transmission, and can interfere with pathogen development in many insects making it an attractive agent for the control of arthropod-borne disease. However, many important species, including Anopheles mosquitoes, are uninfected. Wolbachia can be artificially transferred between insects in the laboratory but this can be a laborious and sometimes fruitless process. We used a simple ex vivo culturing technique to assess the suitability of Wolbachia-host germline associations. Wolbachia infects the dissected germline tissue of multiple insect species when the host tissue and bacteria are cultured together. Ovary and testis infection occurs in a density-dependent manner. Wolbachia strains are more capable of invading the germline of their native or closely related rather than divergent hosts. The ability of Wolbachia to associate with the germline of novel hosts is crucial for the development of stably-transinfected insect lines. Rapid assessment of the suitability of a strain-host combination prior to transinfection may dictate use of a particular Wolbachia strain. Furthermore, the cultured germline tissues of two major Anopheline vectors of Plasmodium parasites are susceptible to Wolbachia infection. This finding further enhances the prospect of using Wolbachia for the biological control of malaria.}, } @article {pmid22558118, year = {2012}, author = {Martinez, J and Duplouy, A and Woolfit, M and Vavre, F and O'Neill, SL and Varaldi, J}, title = {Influence of the virus LbFV and of Wolbachia in a host-parasitoid interaction.}, journal = {PloS one}, volume = {7}, number = {4}, pages = {e35081}, pmid = {22558118}, issn = {1932-6203}, mesh = {Animals ; Drosophila/immunology/*microbiology/*parasitology ; *Host-Pathogen Interactions ; Immunity, Innate/*immunology ; Linear Models ; *Symbiosis ; Viruses/immunology ; Wasps/*virology ; Wolbachia/*physiology ; }, abstract = {Symbionts are widespread and might have a substantial effect on the outcome of interactions between species, such as in host-parasitoid systems. Here, we studied the effects of symbionts on the outcome of host-parasitoid interactions in a four-partner system, consisting of the parasitoid wasp Leptopilina boulardi, its two hosts Drosophila melanogaster and D. simulans, the wasp virus LbFV, and the endosymbiotic bacterium Wolbachia. The virus is known to manipulate the superparasitism behavior of the parasitoid whereas some Wolbachia strains can reproductively manipulate and/or confer pathogen protection to Drosophila hosts. We used two nuclear backgrounds for both Drosophila species, infected with or cured of their respective Wolbachia strains, and offered them to L. boulardi of one nuclear background, either infected or uninfected by the virus. The main defence mechanism against parasitoids, i.e. encapsulation, and other important traits of the interaction were measured. The results showed that virus-infected parasitoids are less frequently encapsulated than uninfected ones. Further experiments showed that this viral effect involved both a direct protective effect against encapsulation and an indirect effect of superparasitism. Additionally, the Wolbachia strain wAu affected the encapsulation ability of its Drosophila host but the direction of this effect was strongly dependent on the presence/absence of LbFV. Our results confirmed the importance of heritable symbionts in the outcome of antagonistic interactions.}, } @article {pmid22548357, year = {2012}, author = {Kraaijeveld, K and Zwanenburg, B and Hubert, B and Vieira, C and De Pater, S and Van Alphen, JJ and Den Dunnen, JT and De Knijff, P}, title = {Transposon proliferation in an asexual parasitoid.}, journal = {Molecular ecology}, volume = {21}, number = {16}, pages = {3898-3906}, doi = {10.1111/j.1365-294X.2012.5582.x}, pmid = {22548357}, issn = {1365-294X}, mesh = {Animals ; Biological Evolution ; DNA Methylation ; DNA Transposable Elements ; Female ; Genome, Insect ; Long Interspersed Nucleotide Elements ; Molecular Sequence Data ; Parthenogenesis/genetics ; Reproduction, Asexual/*genetics ; Terminal Repeat Sequences ; Wasps/microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {The widespread occurrence of sex is one of the most elusive problems in evolutionary biology. Theory predicts that asexual lineages can be driven to extinction by uncontrolled proliferation of vertically transmitted transposable elements (TEs), which accumulate because of the inefficiency of purifying selection in the absence of sex and recombination. To test this prediction, we compared genome-wide TE load between a sexual lineage of the parasitoid wasp Leptopilina clavipes and a lineage of the same species that is rendered asexual by Wolbachia-induced parthenogenesis. We obtained draft genome sequences at 15-20× coverage of both the sexual and the asexual lineages using next-generation sequencing. We identified transposons of most major classes in both lineages. Quantification of TE abundance using coverage depth showed that copy numbers in the asexual lineage exceeded those in the sexual lineage for DNA transposons, but not LTR and LINE-like elements. However, one or a small number of gypsy-like LTR elements exhibited a fourfold higher coverage in the asexual lineage. Quantitative PCR showed that high loads of this gypsy-like TE were characteristic for 11 genetically distinct asexual wasp lineages when compared to sexual lineages. We found no evidence for an overall increase in copy number for all TE types in asexuals as predicted by theory. Instead, we suggest that the expansions of specific TEs are best explained as side effects of (epi)genetic manipulations of the host genome by Wolbachia. Asexuality is achieved in a myriad of ways in nature, many of which could similarly result in TE proliferation.}, } @article {pmid22544247, year = {2012}, author = {Alam, U and Hyseni, C and Symula, RE and Brelsfoard, C and Wu, Y and Kruglov, O and Wang, J and Echodu, R and Alioni, V and Okedi, LM and Caccone, A and Aksoy, S}, title = {Implications of microfauna-host interactions for trypanosome transmission dynamics in Glossina fuscipes fuscipes in Uganda.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {13}, pages = {4627-4637}, pmid = {22544247}, issn = {1098-5336}, support = {R01 AI068932/AI/NIAID NIH HHS/United States ; R03 TW008755/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; Biota ; Host-Pathogen Interactions ; Microbial Interactions ; Trypanosoma/*growth & development ; Trypanosomiasis, African/transmission ; Tsetse Flies/*microbiology/*parasitology ; Uganda ; Viruses/*growth & development ; Wolbachia/*growth & development ; }, abstract = {Tsetse flies (Diptera: Glossinidae) are vectors for African trypanosomes (Euglenozoa: kinetoplastida), protozoan parasites that cause African trypanosomiasis in humans (HAT) and nagana in livestock. In addition to trypanosomes, two symbiotic bacteria (Wigglesworthia glossinidia and Sodalis glossinidius) and two parasitic microbes, Wolbachia and a salivary gland hypertrophy virus (SGHV), have been described in tsetse. Here we determined the prevalence of and coinfection dynamics between Wolbachia, trypanosomes, and SGHV in Glossina fuscipes fuscipes in Uganda over a large geographical scale spanning the range of host genetic and spatial diversity. Using a multivariate analysis approach, we uncovered complex coinfection dynamics between the pathogens and statistically significant associations between host genetic groups and pathogen prevalence. It is important to note that these coinfection dynamics and associations with the host were not apparent by univariate analysis. These associations between host genotype and pathogen are particularly evident for Wolbachia and SGHV where host groups are inversely correlated for Wolbachia and SGHV prevalence. On the other hand, trypanosome infection prevalence is more complex and covaries with the presence of the other two pathogens, highlighting the importance of examining multiple pathogens simultaneously before making generalizations about infection and spatial patterns. It is imperative to note that these novel findings would have been missed if we had employed the standard univariate analysis used in previous studies. Our results are discussed in the context of disease epidemiology and vector control.}, } @article {pmid22540028, year = {2012}, author = {Zeh, JA and Bonilla, MM and Adrian, AJ and Mesfin, S and Zeh, DW}, title = {From father to son: transgenerational effect of tetracycline on sperm viability.}, journal = {Scientific reports}, volume = {2}, number = {}, pages = {375}, pmid = {22540028}, issn = {2045-2322}, mesh = {Animals ; *Arachnida ; *Intergenerational Relations ; Male ; *Paternal Exposure ; Sperm Count ; Spermatozoa/*drug effects ; Tetracycline/*adverse effects ; }, abstract = {The broad-spectrum antibiotic tetracycline is used in animal production, antimicrobial therapy, and for curing arthropods infected with bacterial endosymbionts such as Wolbachia. Tetracycline inhibits mitochondrial translation, and recent evidence indicates that male reproductive traits may be particularly sensitive to this antibiotic. Here, we report the first multi-generation investigation of tetracycline's effects on ejaculate traits. In a study of the pseudoscorpion, Cordylochernes scorpioides, in which siblings were randomly assigned to control and tetracycline treatments across replicate full-sibling families, tetracycline did not affect body size in either sex, female reproduction or sperm number. However, tetracycline-treated males exhibited significantly reduced sperm viability compared to control males, and transmitted this toxic effect of tetracycline on sperm to their untreated sons but not to their F2 grandsons. These results are consistent with tetracycline-induced epigenetic changes in the male germline, and suggest the need for further investigation of transgenerational effects of tetracycline on male reproductive function.}, } @article {pmid22537833, year = {2013}, author = {Balmand, S and Lohs, C and Aksoy, S and Heddi, A}, title = {Tissue distribution and transmission routes for the tsetse fly endosymbionts.}, journal = {Journal of invertebrate pathology}, volume = {112 Suppl}, number = {0}, pages = {S116-22}, pmid = {22537833}, issn = {1096-0805}, support = {R01 AI081774/AI/NIAID NIH HHS/United States ; R01 GM069449/GM/NIGMS NIH HHS/United States ; AI06892/AI/NIAID NIH HHS/United States ; GM069449/GM/NIGMS NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Enterobacteriaceae ; In Situ Hybridization, Fluorescence ; *Symbiosis ; Tsetse Flies/*microbiology ; *Wigglesworthia ; *Wolbachia ; }, abstract = {The tsetse fly Glossina is the vector of the protozoan Trypanosoma brucei spp., which causes Human and Animal African Trypanosomiasis in sub-Saharan African countries. To supplement their unbalanced vertebrate bloodmeal diet, flies permanently harbor the obligate bacterium Wigglesworthia glossinidia, which resides in bacteriocytes in the midgut bacteriome organ as well as in milk gland organ. Tsetse flies also harbor the secondary facultative endosymbionts (S-symbiont) Sodalis glossinidius that infects various tissues and Wolbachia that infects germ cells. Tsetse flies display viviparous reproductive biology where a single embryo hatches and completes its entire larval development in utero and receives nourishments in the form of milk secreted by mother's accessory glands (milk glands). To analyze the precise tissue distribution of the three endosymbiotic bacteria and to infer the way by which each symbiotic partner is transmitted from parent to progeny, we conducted a Fluorescence In situ Hybridization (FISH) study to survey bacterial spatial distribution across the fly tissues. We show that bacteriocytes are mono-infected with Wigglesworthia, while both Wigglesworthia and Sodalis are present in the milk gland lumen. Sodalis was further seen in the uterus, spermathecae, fat body, milk and intracellular in the milk gland cells. Contrary to Wigglesworthia and Sodalis, Wolbachia were the only bacteria infecting oocytes, trophocytes, and embryos at early embryonic stages. Furthermore, Wolbachia were not seen in the milk gland and in the fat body. This work further highlights the diversity of symbiont interactions in multipartner associations and supports two maternal routes of symbiont inheritance in the tsetse fly: Wolbachia through oocytes, and, Wigglesworthia and Sodalis by means of milk gland bacterial infection at early post-embryonic stages.}, } @article {pmid22536103, year = {2012}, author = {Cordaux, R and Pichon, S and Hatira, HB and Doublet, V and Grève, P and Marcadé, I and Braquart-Varnier, C and Souty-Grosset, C and Charfi-Cheikhrouha, F and Bouchon, D}, title = {Widespread Wolbachia infection in terrestrial isopods and other crustaceans.}, journal = {ZooKeys}, volume = {}, number = {176}, pages = {123-131}, pmid = {22536103}, issn = {1313-2970}, abstract = {Wolbachia bacteria are obligate intracellular alpha-Proteobacteria of arthropods and nematodes. Although widespread among isopod crustaceans, they have seldom been found in non-isopod crustacean species. Here, we report Wolbachia infection in fourteen new crustacean species. Our results extend the range of Wolbachia infections in terrestrial isopods and amphipods (class Malacostraca). We report the occurrence of two different Wolbachia strains in two host species (a terrestrial isopod and an amphipod). Moreover, the discovery of Wolbachia in the goose barnacle Lepas anatifera (subclass Thecostraca) establishes Wolbachia infection in class Maxillopoda. The new bacterial strains are closely related to B-supergroup Wolbachia strains previously reported from crustacean hosts. Our results suggest that Wolbachia infection may be much more widespread in crustaceans than previously thought. The presence of related Wolbachia strains in highly divergent crustacean hosts suggests that Wolbachia endosymbionts can naturally adapt to a wide range of crustacean hosts. Given the ability of isopod Wolbachia strains to induce feminization of genetic males or cytoplasmic incompatibility, we speculate that manipulation of crustacean-borne Wolbachia bacteria might represent potential tools for controlling crustacean species of commercial interest and crustacean or insect disease vectors.}, } @article {pmid22534530, year = {2012}, author = {Gravitz, L}, title = {Vector control: The last bite.}, journal = {Nature}, volume = {484}, number = {7395}, pages = {S26-7}, pmid = {22534530}, issn = {1476-4687}, mesh = {Animals ; Beauveria/physiology ; Culicidae/drug effects/microbiology/parasitology/physiology ; Feeding Behavior ; Female ; Insect Vectors/drug effects/microbiology/parasitology/physiology ; Insecticide Resistance ; Insecticide-Treated Bednets/statistics & numerical data ; Malaria/epidemiology/parasitology/*prevention & control/*transmission ; Male ; Mosquito Control/*methods/statistics & numerical data/*trends ; Wolbachia/physiology ; World Health Organization ; }, } @article {pmid22533729, year = {2012}, author = {Zélé, F and Nicot, A and Duron, O and Rivero, A}, title = {Infection with Wolbachia protects mosquitoes against Plasmodium-induced mortality in a natural system.}, journal = {Journal of evolutionary biology}, volume = {25}, number = {7}, pages = {1243-1252}, doi = {10.1111/j.1420-9101.2012.02519.x}, pmid = {22533729}, issn = {1420-9101}, mesh = {Animals ; Culex/*microbiology/*parasitology/physiology ; Disease Vectors ; *Plasmodium ; *Wolbachia ; }, abstract = {In recent years, there has been a shift in the one host-one parasite paradigm with the realization that, in the field, most hosts are coinfected with multiple parasites. Coinfections are particularly relevant when the host is a vector of diseases, because multiple infections can have drastic consequences for parasite transmission at both the ecological and evolutionary timescales. Wolbachia pipientis is the most common parasitic microorganism in insects, and as such, it is of special interest for understanding the role of coinfections in the outcome of parasite infections. Here, we investigate whether Wolbachia can modulate the effect of Plasmodium on what is, arguably, the most important component of the vectorial capacity of mosquitoes: their longevity. For this purpose, and in contrast to recent studies that have focused on mosquito-Plasmodium and/or mosquito-Wolbachia combinations not found in nature, we work on a Wolbachia-mosquito-Plasmodium triad with a common evolutionary history. Our results show that Wolbachia protects mosquitoes from Plasmodium-induced mortality. The results are consistent across two different strains of Wolbachia and repeatable across two different experimental blocks. To our knowledge, this is the first time that such an effect has been shown for Plasmodium-infected mosquitoes and, in particular, in a natural Wolbachia-host combination. We discuss different mechanistic and evolutionary explanations for these results as well as their consequences for Plasmodium transmission.}, } @article {pmid22531297, year = {2012}, author = {Brelsfoard, CL and Dobson, SL}, title = {Population genetic structure of Aedes polynesiensis in the Society Islands of French Polynesia: implications for control using a Wolbachia-based autocidal strategy.}, journal = {Parasites & vectors}, volume = {5}, number = {}, pages = {80}, pmid = {22531297}, issn = {1756-3305}, support = {R01-A1067434//PHS HHS/United States ; }, mesh = {Aedes/classification/genetics/*growth & development/*microbiology ; Animals ; *Biota ; Gene Flow ; Genotype ; Microsatellite Repeats ; Mosquito Control/*methods ; Polynesia ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Aedes polynesiensis is the primary vector of Wuchereria bancrofti in the South Pacific and an important vector of dengue virus. An improved understanding of the mosquito population genetics is needed for insight into the population dynamics and dispersal, which can aid in understanding the epidemiology of disease transmission and control of the vector. In light of the potential release of a Wolbachia infected strain for vector control, our objectives were to investigate the microgeographical and temporal population genetic structure of A. polynesiensis within the Society Islands of French Polynesia, and to compare the genetic background of a laboratory strain intended for release into its population of origin.

METHODS: A panel of eight microsatellite loci were used to genotype A. polynesiensis samples collected in French Polynesia from 2005-2008 and introgressed A. polynesiensis and Aedes riversi laboratory strains. Examination of genetic differentiation was performed using F-statistics, STRUCTURE, and an AMOVA. BAYESASS was used to estimate direction and rates of mosquito movement.

RESULTS: FST values, AMOVA, and STRUCTURE analyses suggest low levels of intra-island differentiation from multiple collection sites on Tahiti, Raiatea, and Maupiti. Significant pair-wise FST values translate to relatively minor levels of inter-island genetic differentiation between more isolated islands and little differentiation between islands with greater commercial traffic (i.e., Tahiti, Raiatea, and Moorea). STRUCTURE analyses also indicate two population groups across the Society Islands, and the genetic makeup of Wolbachia infected strains intended for release is similar to that of wild-type populations from its island of origin, and unlike that of A. riversi.

CONCLUSIONS: The observed panmictic population on Tahiti, Raiatea, and Moorea is consistent with hypothesized gene flow occurring between islands that have relatively high levels of air and maritime traffic, compared to that of the more isolated Maupiti and Tahaa. Gene flow and potential mosquito movement is discussed in relation to trials of applied autocidal strategies.}, } @article {pmid22530989, year = {2012}, author = {McNulty, SN and Mullin, AS and Vaughan, JA and Tkach, VV and Weil, GJ and Fischer, PU}, title = {Comparing the mitochondrial genomes of Wolbachia-dependent and independent filarial nematode species.}, journal = {BMC genomics}, volume = {13}, number = {}, pages = {145}, pmid = {22530989}, issn = {1471-2164}, support = {R03 AI092306/AI/NIAID NIH HHS/United States ; T32-AI007172/AI/NIAID NIH HHS/United States ; R03-AI092306/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Filarioidea/classification/*genetics ; *Genome, Mitochondrial ; Loa/classification/genetics ; Mitochondrial Proteins/genetics/metabolism ; Nucleic Acid Conformation ; Onchocerca/classification/genetics ; Phylogeny ; RNA, Ribosomal/chemistry/metabolism ; RNA, Transfer/chemistry/metabolism ; Wuchereria bancrofti/classification/*genetics ; }, abstract = {BACKGROUND: Many species of filarial nematodes depend on Wolbachia endobacteria to carry out their life cycle. Other species are naturally Wolbachia-free. The biological mechanisms underpinning Wolbachia-dependence and independence in filarial nematodes are not known. Previous studies have indicated that Wolbachia have an impact on mitochondrial gene expression, which may suggest a role in energy metabolism. If Wolbachia can supplement host energy metabolism, reduced mitochondrial function in infected filarial species may account for Wolbachia-dependence. Wolbachia also have a strong influence on mitochondrial evolution due to vertical co-transmission. This could drive alterations in mitochondrial genome sequence in infected species. Comparisons between the mitochondrial genome sequences of Wolbachia-dependent and independent filarial worms may reveal differences indicative of altered mitochondrial function.

RESULTS: The mitochondrial genomes of 5 species of filarial nematodes, Acanthocheilonema viteae, Chandlerella quiscali, Loa loa, Onchocerca flexuosa, and Wuchereria bancrofti, were sequenced, annotated and compared with available mitochondrial genome sequences from Brugia malayi, Dirofilaria immitis, Onchocerca volvulus and Setaria digitata. B. malayi, D. immitis, O. volvulus and W. bancrofti are Wolbachia-dependent while A. viteae, C. quiscali, L. loa, O. flexuosa and S. digitata are Wolbachia-free. The 9 mitochondrial genomes were similar in size and AT content and encoded the same 12 protein-coding genes, 22 tRNAs and 2 rRNAs. Synteny was perfectly preserved in all species except C. quiscali, which had a different order for 5 tRNA genes. Protein-coding genes were expressed at the RNA level in all examined species. In phylogenetic trees based on mitochondrial protein-coding sequences, species did not cluster according to Wolbachia dependence.

CONCLUSIONS: Thus far, no discernable differences were detected between the mitochondrial genome sequences of Wolbachia-dependent and independent species. Additional research will be needed to determine whether mitochondria from Wolbachia-dependent filarial species show reduced function in comparison to the mitochondria of Wolbachia-independent species despite their sequence-level similarities.}, } @article {pmid22530696, year = {2012}, author = {Andersen, SB and Boye, M and Nash, DR and Boomsma, JJ}, title = {Dynamic Wolbachia prevalence in Acromyrmex leaf-cutting ants: potential for a nutritional symbiosis.}, journal = {Journal of evolutionary biology}, volume = {25}, number = {7}, pages = {1340-1350}, doi = {10.1111/j.1420-9101.2012.02521.x}, pmid = {22530696}, issn = {1420-9101}, mesh = {Animals ; Ants/growth & development/*microbiology/physiology ; In Situ Hybridization, Fluorescence ; Larva/microbiology/physiology ; Pupa/microbiology/physiology ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {Wolbachia are renowned as reproductive parasites, but their phenotypic effects in eusocial insects are not well understood. We used a combination of qrt-PCR, fluorescence in situ hybridization and laser scanning confocal microscopy to evaluate the dynamics of Wolbachia infections in the leaf-cutting ant Acromyrmex octospinosus across developmental stages of sterile workers. We confirm that workers are infected with one or two widespread wsp genotypes of Wolbachia, show that colony prevalence is always 100% and characterize two rare recombinant genotypes. One dominant genotype is always present and most abundant, whereas another only proliferates in adult workers of some colonies and is barely detectable in larvae and pupae. An explanation may be that Wolbachia genotypes compete for host resources in immature stages while adult tissues provide substantially more niche space. Tissue-specific prevalence of the two genotypes differs, with the rarer genotype being over-represented in the adult foregut and thorax muscles. Both genotypes occur extracellularly in the foregut, suggesting an unknown mutualistic function in worker ant nutrition. Both genotypes are also abundant in the faecal fluid of the ants, suggesting that they may have extended functional phenotypes in the fungus garden that the ants manure with their own faeces.}, } @article {pmid22530444, year = {2012}, author = {Behbahani, A}, title = {Wolbachia infection and mitochondrial DNA comparisons among Culex mosquitoes in South West Iran.}, journal = {Pakistan journal of biological sciences : PJBS}, volume = {15}, number = {1}, pages = {54-57}, doi = {10.3923/pjbs.2012.54.57}, pmid = {22530444}, issn = {1028-8880}, mesh = {Animals ; Culex/cytology/*genetics/*microbiology ; DNA, Mitochondrial/*genetics ; Haplotypes ; Insect Vectors/cytology/genetics/microbiology ; Iran ; Wolbachia/genetics/*pathogenicity ; }, abstract = {The control of mosquito borne diseases needs new methods given widespread insecticide resistance in many mosquito species. The inherited endosymbiont Wolbachia, found in many arthropods, provides a biological system to reduce the transmission of these diseases and replace the population of vectors with non-vectors using cytoplasmic incompatibility. The aim of this study was to understand the rate of Wolbachia infection among Culex species in the region and to see the effect of Wolbachia infection on mitochondrial genome. In this study three species of Culex mosquitoes were collected from Shoushtar in south west of Iran and examined for Wolbachia infection by Polymerase Chain Reaction (PCR). All of the C. quinquefasciatus specimens were infected with Wolbachia, while C. tritaeniorynchus and C. theileri showed no infection with Wolbachia. The 340 bp of AT rich of mtDNA was sequenced from 30 individuals, 10 individuals of each species. Three sequence haplotypes were found in C. tritaeniorynchus and C. theileri while there was only one haplotype in C. quinquefasciatus. The reduction of haplotypes diversity may be result of a sweep of Wolbachia in this species.}, } @article {pmid22529882, year = {2012}, author = {Fagen, JR and Giongo, A and Brown, CT and Davis-Richardson, AG and Gano, KA and Triplett, EW}, title = {Characterization of the Relative Abundance of the Citrus Pathogen Ca. Liberibacter Asiaticus in the Microbiome of Its Insect Vector, Diaphorina citri, using High Throughput 16S rRNA Sequencing.}, journal = {The open microbiology journal}, volume = {6}, number = {}, pages = {29-33}, pmid = {22529882}, issn = {1874-2858}, abstract = {The relationship between the causal agent of Huanglongbing (HLB), Ca. Liberibacter asiaticus(Las), and the naturally occurring endosymbiotic community of its insect vector, the Asian citrus psyllid (ACP), Diaphorina citri, was studied. Variation was observed in the titer of Las within an ACP population feeding on the same material. The cause of this disparity is unknown, and has implications for Las transmission and the spread of HLB. This study utilizes culture independent methods to establish the relationship between the ACP's microbial community and Las acquisition. DNA from 21 psyllids was amplified using universal 16S rRNA primers with Illumina adaptor regions and a sample-specific 7- base identifier. These amplicons were then batch-sequenced on the Illumina platform. The resulting sequences were separated by the identifier, and compared to known sequences in a 16S rRNA database. The microbial communities of each psyllid were compared to determine whether a correlation exists between the ACP's endosymbionts and level of Las acquisition.ACPs were dominated by the same four bacterialgenera regardless of the abundance of Ca.Liberibacter. A combination of qPCR and Illumina sequencing was used to establish an infection gradient among the sampled ACPs. The Ca. Liberibacter titer within the insect was found to have a strong negative relationship with an endosymbiont residing in the syncytium of the mycetocyte and a positive relationship with Wolbachia. These correlations have implications in the acquisition of Las by the ACP as well as the activities of Las within this vector.}, } @article {pmid22525064, year = {2012}, author = {Hail, D and Dowd, SE and Bextine, B}, title = {Identification and location of symbionts associated with potato psyllid (Bactericera cockerelli) lifestages.}, journal = {Environmental entomology}, volume = {41}, number = {1}, pages = {98-107}, doi = {10.1603/EN11198}, pmid = {22525064}, issn = {1938-2936}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; Hemiptera/*microbiology ; *Metagenome ; Nymph/microbiology ; Ovum/microbiology ; Pest Control, Biological ; Plant Diseases/microbiology ; Polymerase Chain Reaction ; Principal Component Analysis ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/analysis ; Sequence Analysis, DNA ; Symbiosis ; }, abstract = {The potato psyllid (Bactericera cockerelli, Sulc) is an invasive pest of solenaceous plants including potatoes (Solanum tuberosum L.)and tomatoes (Solanum lycopersicum L.). The insect transmits the phytopathogen Candidatus Liberibacter solanacearum, which has been identified as the causal agent of Zebra Chip in potatoes. The microbiome of the potato psyllid provides knowledge of the insect's bacterial makeup which enables researchers to develop targeted biological control strategies. In this study, the microbes associated with four B. cockerelli life stages were evaluated by 16S bTEFAP pyrosequencing. The sequences were compared with a 16S-rDNA database derived from NCBI's GenBank. Some bacteria identified are initial discoveries. Species of Wolbachia, Rhizobium, Gordonia, Mycobacterium, Xanthomonas and others were also detected and an assessment of the microbiome associated with B. cockerelli was established.}, } @article {pmid22522691, year = {2012}, author = {Lee, SF and White, VL and Weeks, AR and Hoffmann, AA and Endersby, NM}, title = {High-throughput PCR assays to monitor Wolbachia infection in the dengue mosquito (Aedes aegypti) and Drosophila simulans.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {13}, pages = {4740-4743}, pmid = {22522691}, issn = {1098-5336}, mesh = {Aedes/*microbiology ; Animals ; Drosophila/*microbiology ; Fluorescence ; High-Throughput Screening Assays/economics/*methods ; Polymerase Chain Reaction/economics/*methods ; Sensitivity and Specificity ; Wolbachia/*genetics/*isolation & purification ; }, abstract = {We have developed and validated two new fluorescence-based PCR assays to detect the Wolbachia wMel strain in Aedes aegypti and the wRi and wAu strains in Drosophila simulans. The new assays are accurate, informative, and cost-efficient for large-scale Wolbachia screening.}, } @article {pmid22516306, year = {2013}, author = {Schneider, DI and Garschall, KI and Parker, AG and Abd-Alla, AM and Miller, WJ}, title = {Global Wolbachia prevalence, titer fluctuations and their potential of causing cytoplasmic incompatibilities in tsetse flies and hybrids of Glossina morsitans subgroup species.}, journal = {Journal of invertebrate pathology}, volume = {112 Suppl}, number = {Suppl 1}, pages = {S104-15}, pmid = {22516306}, issn = {1096-0805}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; DNA Fingerprinting ; Insect Vectors/microbiology ; Minisatellite Repeats/*genetics ; Polymerase Chain Reaction/methods ; Prevalence ; Tsetse Flies/*microbiology ; Wolbachia/*genetics ; }, abstract = {We demonstrate the high applicability of a novel VNTR-based (Variable-Number-Tandem-Repeat) molecular screening tool for fingerprinting Wolbachia-infections in tsetse flies. The VNTR-141 locus provides reliable and concise differentiation between Wolbachia strains deriving from Glossina morsitans morsitans, Glossina morsitans centralis, and Glossina brevipalpis. Moreover, we show that certain Wolbachia-infections in Glossina spp. are capable of escaping standard PCR screening methods by 'hiding' as low-titer infections below the detection threshold. By applying a highly sensitive PCR-blot technique to our Glossina specimen, we were able to enhance the symbiont detection limit substantially and, consequently, trace unequivocally Wolbachia-infections at high prevalence in laboratory-reared G. swynnertoni individuals. To our knowledge, Wolbachia-persistence was reported exclusively for field-collected samples, and at low prevalence only. Finally, we highlight the substantially higher Wolbachia titer levels found in hybrid Glossina compared to non-hybrid hosts and the possible impact of these titers on hybrid host fitness that potentially trigger incipient speciation in tsetse flies.}, } @article {pmid22504806, year = {2012}, author = {Matsuura, Y and Kikuchi, Y and Meng, XY and Koga, R and Fukatsu, T}, title = {Novel clade of alphaproteobacterial endosymbionts associated with stinkbugs and other arthropods.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {12}, pages = {4149-4156}, pmid = {22504806}, issn = {1098-5336}, mesh = {Alphaproteobacteria/*classification/genetics/*isolation & purification/physiology ; Animals ; Arthropods/*microbiology ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Here we report a novel clade of secondary endosymbionts associated with insects and other arthropods. Seed bugs of the genus Nysius (Hemiptera: Lygaeidae) harbor the primary gammaproteobacterial symbiont Schneideria nysicola within a pair of bacteriomes in the abdomen. Our survey of Nysius species for their facultative bacterial associates consistently yielded a novel type of alphaproteobacterial 16S rRNA gene sequence in addition to those of Wolbachia. Diagnostic PCR survey of 343 individuals representing 24 populations of four Nysius species revealed overall detection rates of the alphaproteobacteria at 77.6% in Nysius plebeius, 87.7% in Nysius sp. 1, 81.0% in Nysius sp. 2, and 100% in Nysius expressus. Further survey of diverse stinkbugs representing 24 families, 191 species, and 582 individuals detected the alphaproteobacteria from an additional 12 species representing six families. Molecular phylogenetic analysis showed that the alphaproteobacteria from the stinkbugs form a distinct and coherent monophyletic group in the order Rickettsiales together with several uncharacterized endosymbionts from fleas and ticks. The alphaproteobacterial symbiont clade was allied to bacterial clades such as the endosymbionts of acanthamoebae, the endosymbionts of cnidarians, and Midichloria spp., the mitochondrion-associated endosymbionts of ticks. In situ hybridization and electron microscopy identified small filamentous bacterial cells in various tissues of N. plebeius, including the bacteriome and ovary. The concentrated localization of the symbiont cells at the anterior pole of oocytes indicated its vertical transmission route through host insect generations. The designation "Candidatus Lariskella arthropodarum" is proposed for the endosymbiont clade.}, } @article {pmid22504801, year = {2012}, author = {Salunke, BK and Salunkhe, RC and Dhotre, DP and Walujkar, SA and Khandagale, AB and Chaudhari, R and Chandode, RK and Ghate, HV and Patole, MS and Werren, JH and Shouche, YS}, title = {Determination of Wolbachia diversity in butterflies from Western Ghats, India, by a multigene approach.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {12}, pages = {4458-4467}, pmid = {22504801}, issn = {1098-5336}, mesh = {Animals ; Butterflies/*microbiology ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; *Genetic Variation ; Genotype ; India ; Molecular Sequence Data ; *Multilocus Sequence Typing ; Recombination, Genetic ; Wolbachia/*classification/*genetics ; }, abstract = {Members of the genus Wolbachia are intracellular bacteria that are widespread in arthropods and establish diverse symbiotic associations with their hosts, ranging from mutualism to parasitism. Here we present the first detailed analyses of Wolbachia in butterflies from India with screening of 56 species. Twenty-nine species (52%) representing five families were positive for Wolbachia. This is the first report of Wolbachia infection in 27 of the 29 species; the other two were reported previously. This study also provides the first evidence of infection in the family Papilionidae. A striking diversity was observed among Wolbachia strains in butterfly hosts based on five multilocus sequence typing (MLST) genes, with 15 different sequence types (STs). Thirteen STs are new to the MLST database, whereas ST41 and ST125 were reported earlier. Some of the same host species from this study carried distinctly different Wolbachia strains, whereas the same or different butterfly hosts also harbored closely related Wolbachia strains. Butterfly-associated STs in the Indian sample originated by recombination and point mutation, further supporting the role of both processes in generating Wolbachia diversity. Recombination was detected only among the STs in this study and not in those from the MLST database. Most of the strains were remarkably similar in their wsp genotype, despite divergence in MLST. Only two wsp alleles were found among 25 individuals with complete hypervariable region (HVR) peptide profiles. Although both wsp and MLST show variability, MLST gives better separation between the strains. Completely different STs were characterized for the individuals sharing the same wsp alleles.}, } @article {pmid22504433, year = {2012}, author = {Wu, K and Hoy, MA}, title = {Cardinium is associated with reproductive incompatibility in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae).}, journal = {Journal of invertebrate pathology}, volume = {110}, number = {3}, pages = {359-365}, doi = {10.1016/j.jip.2012.03.027}, pmid = {22504433}, issn = {1096-0805}, mesh = {Animals ; Bacterial Infections/microbiology/*veterinary ; Bacteroidetes/*physiology ; Breeding ; Crosses, Genetic ; Female ; Fertility/physiology ; Host-Pathogen Interactions/*physiology ; Longevity ; Male ; Mites/*microbiology/physiology ; Pest Control, Biological ; Reproduction/*physiology ; Sex Factors ; Symbiosis/physiology ; }, abstract = {Cardinium, a bacterium from the Bacteroidetes group, is associated with reproductive manipulations such as cytoplasmic incompatibility, parthenogenesis, and feminization in some arthropod species. We, and others, have shown that Cardinium, but not Wolbachia, is an endosymbiont in some populations of Metaseiulus occidentalis, a phytoseiid mite that is an important predator of spider mite pests of agricultural crops. However, the precise biological effects that Cardinium may have on M. occidentalis remain unclear. In this study we show, in two sets of crosses between different colonies of Cardinium-free (C-) M. occidentalis females and Cardinium-containing (C+) males, that fecundity was reduced in parental females, F1 progeny survival rates were reduced, and fewer female progeny were produced when compared to the reciprocal and control crosses. There were no differences in these attributes in the reciprocal and two control crosses. Cardinium was transmitted maternally but there was no observed paternal transmission. Finally, Cardinium did not cause asexual (thelytoky) reproduction in M. occidentalis. Thus, Cardinium is associated with nonreciprocal reproductive incompatibility in M. occidentalis and our results support the hypothesis that Cardinium is a reproductive parasite in this agriculturally important predator. Cardinium may therefore affect the evolution and ecology of M. occidentalis and biological control efforts using this mite.}, } @article {pmid22500583, year = {2012}, author = {Caragata, EP and Walker, T}, title = {Using bacteria to treat diseases.}, journal = {Expert opinion on biological therapy}, volume = {12}, number = {6}, pages = {701-712}, doi = {10.1517/14712598.2012.677429}, pmid = {22500583}, issn = {1744-7682}, mesh = {Animals ; Culicidae/growth & development/*microbiology/*parasitology ; Dengue/epidemiology/parasitology/*prevention & control/transmission ; Humans ; *Insect Vectors ; Larva/microbiology/parasitology ; Malaria/epidemiology/parasitology/*prevention & control/transmission ; Mosquito Control/*methods ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {INTRODUCTION: Mosquito-borne diseases such as malaria and dengue fever result in significant morbidity and mortality in developing countries. Vector control is often the most effective strategy to prevent disease transmission and novel methods are required to complement existing insecticide-based strategies. Biological control uses natural predators or pathogens to kill mosquitoes or reduce their capacity to transmit disease. Bacteria such as Wolbachia have been proposed to have the potential to provide effective biological control of mosquitoes.

AREAS COVERED: A review of the potential role of bacteria in the control of mosquito-borne diseases highlighting the advantages and disadvantages of each strategy. In particular, a comprehensive summary of the progress made using the bacterial endosymbiont Wolbachia for dengue control.

EXPERT OPINION: Pathogenic bacteria such as Bti can be used to kill mosquito larvae and several endosymbiotic bacteria such as Asaia could be genetically transformed to alter the mosquito's ability to transmit pathogens. The endosymbiotic bacterium Wolbachia has been successfully introduced into the principal vector of dengue, Aedes aegypti, and induces a variety of phenotypic effects that are predicted to reduce dengue transmission. The release of Wolbachia-infected mosquitoes has been undertaken as part of preliminary trials to determine the applied use of this bacterium for mosquito-borne disease control.}, } @article {pmid22497736, year = {2012}, author = {Pichon, S and Bouchon, D and Liu, C and Chen, L and Garrett, RA and Grève, P}, title = {The expression of one ankyrin pk2 allele of the WO prophage is correlated with the Wolbachia feminizing effect in isopods.}, journal = {BMC microbiology}, volume = {12}, number = {}, pages = {55}, pmid = {22497736}, issn = {1471-2180}, mesh = {Alleles ; Animals ; Ankyrins/*genetics ; Female ; Gene Expression Profiling ; *Gene Expression Regulation, Viral ; Isopoda/*microbiology/physiology ; Male ; Molecular Sequence Data ; Prophages/*genetics ; Sequence Analysis, DNA ; Sex Characteristics ; Viral Proteins/*biosynthesis/genetics ; Wolbachia/*virology ; }, abstract = {BACKGROUND: The maternally inherited α-Proteobacteria Wolbachia pipientis is an obligate endosymbiont of nematodes and arthropods, in which they induce a variety of reproductive alterations, including Cytoplasmic Incompatibility (CI) and feminization. The genome of the feminizing wVulC Wolbachia strain harboured by the isopod Armadillidium vulgare has been sequenced and is now at the final assembly step. It contains an unusually high number of ankyrin motif-containing genes, two of which are homologous to the phage-related pk1 and pk2 genes thought to contribute to the CI phenotype in Culex pipiens. These genes encode putative bacterial effectors mediating Wolbachia-host protein-protein interactions via their ankyrin motifs.

RESULTS: To test whether these Wolbachia homologs are potentially involved in altering terrestrial isopod reproduction, we determined the distribution and expression of both pk1 and pk2 genes in the 3 Wolbachia strains that induce CI and in 5 inducing feminization of their isopod hosts. Aside from the genes being highly conserved, we found a substantial copy number variation among strains, and that is linked to prophage diversity. Transcriptional analyses revealed expression of one pk2 allele (pk2b2) only in the feminizing Wolbachia strains of isopods.

CONCLUSIONS: These results reveal the need to investigate the functions of Wolbachia ankyrin gene products, in particular those of Pk2, and their host targets with respect to host sex manipulation.}, } @article {pmid22494817, year = {2012}, author = {Vavre, F and Charlat, S}, title = {Making (good) use of Wolbachia: what the models say.}, journal = {Current opinion in microbiology}, volume = {15}, number = {3}, pages = {263-268}, doi = {10.1016/j.mib.2012.03.005}, pmid = {22494817}, issn = {1879-0364}, mesh = {Animals ; Arthropods ; *Biological Control Agents ; Biological Evolution ; Female ; Filarioidea ; Host-Pathogen Interactions ; Male ; *Models, Biological ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia, probably the most common animal endosymbiont, infects a wide range of arthropods as well as filarial nematodes. Generally vertically transmitted from mothers to offspring, it has evolved various strategies, ranging from brutal male-killing to mutualism, which facilitate invasion and persistence of the infections within host populations. Current interest in Wolbachia as a potential control agent against harmful nematodes and arthropods makes it important to be able to predict Wolbachia epidemiology and evolutionary trajectory. Here we highlight recent theoretical developments and suggest future modelling and empirical directions for basic and applied research in this domain.}, } @article {pmid22468989, year = {2012}, author = {Dhami, MK and Turner, AP and Deines, P and Beggs, JR and Taylor, MW}, title = {Ultrastructural and molecular characterization of a bacterial symbiosis in the ecologically important scale insect family Coelostomidiidae.}, journal = {FEMS microbiology ecology}, volume = {81}, number = {3}, pages = {537-546}, doi = {10.1111/j.1574-6941.2012.01378.x}, pmid = {22468989}, issn = {1574-6941}, mesh = {Animals ; Bacteroidetes/classification/genetics/*isolation & purification/physiology ; Female ; Gammaproteobacteria/classification/genetics/*isolation & purification/physiology ; Hemiptera/*microbiology/physiology ; In Situ Hybridization, Fluorescence ; New Zealand ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Scale insects are important ecologically and as agricultural pests. The majority of scale insect taxa feed exclusively on plant phloem sap, which is carbon rich but deficient in essential amino acids. This suggests that, as seen in the related aphids and psyllids, scale insect nutrition might also depend upon bacterial symbionts, yet very little is known about scale insect-bacteria symbioses. We report here the first identification and molecular characterization of symbiotic bacteria associated with the New Zealand giant scale Coelostomidia wairoensis, using fluorescence in situ hybridization (FISH), transmission electron microscopy (TEM) and 16S rRNA gene-based analysis. Dissection and FISH confirmed the location of the bacteria in large, paired, multilobate organs in the abdominal region of the insect. TEM indicated that the dominant pleomorphic bacteria were confined to bacteriocytes in the sheath-enclosed bacteriome. Phylogenetic analysis revealed the presence of three distinct bacterial types, the bacteriome-associated B-symbiont (Bacteroidetes), an Erwinia-related symbiont (Gammaproteobacteria) and Wolbachia sp. (Alphaproteobacteria). This study extends the current knowledge of scale insect symbionts and is the first microbiological investigation of the ecologically important coelostomidiid scales.}, } @article {pmid22465193, year = {2012}, author = {Ricci, I and Damiani, C and Capone, A and DeFreece, C and Rossi, P and Favia, G}, title = {Mosquito/microbiota interactions: from complex relationships to biotechnological perspectives.}, journal = {Current opinion in microbiology}, volume = {15}, number = {3}, pages = {278-284}, doi = {10.1016/j.mib.2012.03.004}, pmid = {22465193}, issn = {1879-0364}, mesh = {Animals ; Biotechnology ; Culicidae/*microbiology/*physiology ; Host-Pathogen Interactions ; Insect Vectors/*microbiology/*physiology ; Metagenome/physiology ; Wolbachia/physiology ; Yeasts/physiology ; }, abstract = {To date around 3500 different species of mosquito have been described, several tens of which are vectors of pathogens of remarkable interest in public health. Mosquitoes are present all around the world showing a great ability to adapt to very different types of habitats where they play relevant ecological roles. It is very likely that components of the mosquito microbiota have given the mosquito a great capacity to adapt to different environments. Current advances in understanding the mosquito-microbiota relationships may have a great impact in a better understanding of some traits of mosquito biology and in the development of innovative mosquito-borne disease-control strategies aimed to reduce mosquito vectorial capacity and/or inhibiting pathogen transmission.}, } @article {pmid24031883, year = {2012}, author = {Zimmermann, BL and Almerão, MP and Bouchon, D and Araujo, PB}, title = {Detection of Wolbachia (Alphaproteobacteria: rickettsiales) in three species of terrestrial isopods (crustacea: isopoda: oniscidea) in Brazil.}, journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]}, volume = {43}, number = {2}, pages = {711-715}, pmid = {24031883}, issn = {1517-8382}, abstract = {Terrestrial isopods are widely infected with Wolbachia. However, little is known about the presence of bacteria in the Neotropical species. The objective of this study was to test the hypothesis of presence of Wolbachia infection in the native species of terrestrial isopods, Atlantoscia floridana and Circoniscus bezzii, and in the introduced species Burmoniscus meeusei.}, } @article {pmid22450512, year = {2012}, author = {Watanabe, M and Tagami, Y and Miura, K and Kageyama, D and Stouthamer, R}, title = {Distribution patterns of Wolbachia endosymbionts in the closely related flower bugs of the genus Orius: implications for coevolution and horizontal transfer.}, journal = {Microbial ecology}, volume = {64}, number = {2}, pages = {537-545}, pmid = {22450512}, issn = {1432-184X}, mesh = {Animals ; *Biological Evolution ; Cloning, Molecular ; Flowers/*parasitology ; *Gene Transfer, Horizontal ; Heteroptera/classification/*microbiology ; Japan ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Wolbachia/classification/*genetics/physiology ; }, abstract = {Vertical transmission is the primary route of the endosymbiont Wolbachia for its own spread among invertebrate hosts, but horizontal transmission between different hosts is believed to have occurred multiple times. However, it is not well known how Wolbachia commonly spread among closely related hosts. We focused on the closely related species of the minute pirate bugs belonging to the genus Orius, which are important biological control agents in agricultural crops because they are the most useful natural enemy of various tiny pests, such as thrips. Here, we examined five Orius species (Orius sauteri, Orius nagaii, Orius minutus, Orius strigicollis, and Orius tantillus) from eight geographic localities in Japan for Wolbachia infection. Two distinct strains, wOus1 and wOus2, were detected based on Wolbachia surface protein (wsp) gene sequencing. Furthermore, multilocus sequence typing revealed that each of the strains comprised two variants that differed in a single nucleotide. The overall distribution patterns of the two Wolbachia strains were found to differ among host species: prevalent double infection with wOus1 and wOus2 in O. strigicollis; fixation of single infection with wOus2 in O. nagaii; occurrence of single infection with wOus1 in O. sauteri; prevalence of single infection with wOus1 in O. minutus with an exception in a single population; and lack of Wolbachia infection in O. tantillus. Such differences in the distribution patterns of Wolbachia may reflect the evolutionary history of Wolbachia infection among Orius species and/or ecological and physiological differences among the Orius species that determine the invasiveness and maintenance of the two Wolbachia strains.}, } @article {pmid22448745, year = {2012}, author = {Sekeyová, Z and Mediannikov, O and Roux, V and Subramanian, G and Spitalská, E and Kristofík, J and Darolová, A and Raoult, D}, title = {Identification of Rickettsia africae and Wolbachia sp. in Ceratophyllus garei fleas from Passerine birds migrated from Africa.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {12}, number = {7}, pages = {539-543}, pmid = {22448745}, issn = {1557-7759}, mesh = {Africa ; Animal Migration ; Animals ; Bird Diseases/*parasitology/transmission ; DNA, Bacterial/genetics ; Europe ; Flea Infestations/parasitology/transmission/veterinary ; Genes, Bacterial/genetics ; Insect Vectors/*microbiology ; Phylogeny ; RNA, Bacterial/genetics ; Rickettsia/genetics/*isolation & purification ; Rickettsia Infections/transmission ; Rickettsiaceae Infections/transmission ; Siphonaptera/*microbiology ; Slovakia ; Songbirds/*parasitology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The aim of the study was to reveal new aspects of the role of flea vector taken from migratory birds by screening of specimens with molecular biological methods. A field study was done in fishponds in Slovakia. Actually, 47 fleas were collected from reed warblers (Acrocephalus scirpaceus) and their nests. DNA was extracted and analyzed for representatives of the orders Rickettsiales. A rickettsia that shares 99.7% of identity by gltA gene with Rickettsia africae was identified in Ceratophyllus garei collected from A. scirpaceus. Moreover, two Wolbachia sp. were also detected in fleas. This is the first record of R. africae and Wolbachia sp. identified so far in Central Europe in fleas collected from migratory bird returning from Africa. This molecular study extends the geographic range and vector spectrum of arthropod-borne agents.}, } @article {pmid22447527, year = {2011}, author = {Drosopoulou, E and Augustinos, AA and Nakou, I and Koeppler, K and Kounatidis, I and Vogt, H and Papadopoulos, NT and Bourtzis, K and Mavragani-Tsipidou, P}, title = {Genetic and cytogenetic analysis of the American cherry fruit fly, Rhagoletis cingulata (Diptera: Tephritidae).}, journal = {Genetica}, volume = {139}, number = {11-12}, pages = {1449-1464}, pmid = {22447527}, issn = {1573-6857}, mesh = {Alleles ; Animals ; Chromosomes, Insect ; Cytogenetic Analysis ; Female ; Genes, Insect ; Karyotype ; Male ; Microsatellite Repeats ; Mitosis ; Phylogeny ; Polytene Chromosomes ; Sex Chromosomes ; Tephritidae/*genetics ; }, abstract = {The American eastern cherry fruit fly, Rhagoletis cingulata, a pest of cherries in the western hemisphere, invaded Europe in 1983, and since then dispersed to several European countries. Information on the genetics and cytogenetics of this pest is very scarce. The mitotic karyotype and detailed photographic maps of the salivary gland polytene chromosomes of R. cingulata are presented here. The mitotic metaphase complement consists of six pairs of chromosomes with the sex chromosomes being very small and similar in size. The analysis of the salivary gland polytene complement shows a total number of five long chromosomes (10 polytene arms), which correspond to the five autosomes of the mitotic nuclei and an extrachromosomal heterochromatic mass, which corresponds to the sex chromosomes. The banding patterns and the most characteristic features and prominent landmarks of each polytene chromosome are presented and discussed. Chromosomal homologies between R. cingulata, R. completa and R. cerasi are also proposed, based on the comparison of chromosome banding patterns. Furthermore, the detection and characterization of Wolbachia pipientis in the R. cingulata population studied is presented and the potential correlation with the asynaptic phenomena found in its polytene complement is discussed. In addition, 10 out of 24 microsatellite markers developed for other Rhagoletis species are cross-amplified, evaluated and proposed as useful markers for population and genetic studies in R. cingulata.}, } @article {pmid22433886, year = {2011}, author = {Sunish, IP and Rajendran, R and Paramasivan, R and Dhananjeyan, KJ and Tyagi, BK}, title = {Wolbachia endobacteria in a natural population of Culex quinquefasciatus from filariasis endemic villages of south India and its phylogenetic implication.}, journal = {Tropical biomedicine}, volume = {28}, number = {3}, pages = {569-576}, pmid = {22433886}, issn = {2521-9855}, mesh = {Animals ; Culex/*microbiology ; DNA, Bacterial/chemistry/genetics ; Endemic Diseases ; Female ; Filariasis/epidemiology ; Humans ; India/epidemiology ; Male ; *Phylogeny ; Prevalence ; Sequence Analysis, DNA ; Sequence Homology ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Understanding Wolbachia mosquito interactions have been recognized as an important concept to develop novel vector control strategies. The prevalence of Wolbachia endobacteria in a natural population of the filariasis vector Culex quinquefasciatus was determined by the polymerase chain reaction method. Earlier workers had estimated the infection rates of Wolbachia with only one or very few individuals per species. In our study large number of specimens were assayed, and a total of 750 adult Culex quinquefasciatus mosquitoes were collected from three south Indian villages of Tirukoilur and Mugaiyur blocks, monthly for a period of five months (December 2006 to April 2007) and screened for the presence of Wolbachia. The percentage prevalence in adult males ranged from 88% to 96%; while in females from 84% to 100%. An overall prevalence of 91.2% was observed. There was no significant difference observed in the proportion of mosquitoes positive for Wolbachia between males and females, and also between different months of the survey; except during the month of February '07. The wsp gene sequence of the Wolbachia strain of Cx. quinquefasciatus detected was BLAST analysed and showed 99% sequence similarity with Wolbachia sp. of Culex pipiens isolated from different geographical regions. Phylogenetic analysis based on wsp gene fragments showed that the present Wolbachia isolate was closely related with Wolbachia from Culex pipens pipiens, Niphotettix virescens (Order: Hemiptera) and Cnaphalocrosis medinalis (Order: Lepidoptera).}, } @article {pmid22433115, year = {2012}, author = {Zouache, K and Michelland, RJ and Failloux, AB and Grundmann, GL and Mavingui, P}, title = {Chikungunya virus impacts the diversity of symbiotic bacteria in mosquito vector.}, journal = {Molecular ecology}, volume = {21}, number = {9}, pages = {2297-2309}, doi = {10.1111/j.1365-294X.2012.05526.x}, pmid = {22433115}, issn = {1365-294X}, mesh = {Aedes/*microbiology/*virology ; Alphaproteobacteria/physiology ; Animals ; *Bacterial Physiological Phenomena ; Bacteroidetes/physiology ; Biodiversity ; Chikungunya virus/genetics/*physiology ; Enterobacteriaceae/physiology ; Gammaproteobacteria/physiology ; Host-Pathogen Interactions ; Insect Vectors/*microbiology/*virology ; *Symbiosis ; Virus Replication ; Wolbachia/physiology ; }, abstract = {Mosquitoes transmit numerous arboviruses including dengue and chikungunya virus (CHIKV). In recent years, mosquito species Aedes albopictus has expanded in the Indian Ocean region and was the principal vector of chikungunya outbreaks in La Reunion and neighbouring islands in 2005 and 2006. Vector-associated bacteria have recently been found to interact with transmitted pathogens. For instance, Wolbachia modulates the replication of viruses or parasites. However, there has been no systematic evaluation of the diversity of the entire bacterial populations within mosquito individuals particularly in relation to virus invasion. Here, we investigated the effect of CHIKV infection on the whole bacterial community of Ae. albopictus. Taxonomic microarrays and quantitative PCR showed that members of Alpha- and Gammaproteobacteria phyla, as well as Bacteroidetes, responded to CHIKV infection. The abundance of bacteria from the Enterobacteriaceae family increased with CHIKV infection, whereas the abundance of known insect endosymbionts like Wolbachia and Blattabacterium decreased. Our results clearly link the pathogen propagation with changes in the dynamics of the bacterial community, suggesting that cooperation or competition occurs within the host, which may in turn affect the mosquito traits like vector competence.}, } @article {pmid22433114, year = {2012}, author = {Tamarozzi, F and Tendongfor, N and Enyong, PA and Esum, M and Faragher, B and Wanji, S and Taylor, MJ}, title = {Long term impact of large scale community-directed delivery of doxycycline for the treatment of onchocerciasis.}, journal = {Parasites & vectors}, volume = {5}, number = {}, pages = {53}, pmid = {22433114}, issn = {1756-3305}, mesh = {Adult ; Aged ; Aged, 80 and over ; Animals ; Anti-Bacterial Agents/*administration & dosage ; Biopsy ; Cameroon/epidemiology ; Doxycycline/*administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Onchocerca volvulus/*microbiology ; Onchocerciasis/epidemiology/parasitology/*prevention & control ; Skin/parasitology ; Treatment Outcome ; Wolbachia/*drug effects ; }, abstract = {BACKGROUND: Anti-Wolbachia treatment with doxycycline is effective in sterilising and killing adult Onchocerca volvulus nematodes, proving superior to ivermectin and of great potential as an alternative approach for the treatment and control of onchocerciasis, particularly in areas of Loa loa co-endemicity. Nevertheless, the length of the required treatment poses potential logistical problems and risk of poor compliance, raising a barrier to the use of doxycycline in Mass Drug Administration (MDA) strategies. In 2007 and 2008 a feasibility trial of community-directed treatment with doxycycline was carried out in two health districts in Cameroon, co-endemic for O. volvulus and L. loa. With 17,519 eligible subjects, the therapeutic coverage was 73.8% with 97.5% compliance, encouraging the feasibility of using doxycycline community-directed delivery in restricted populations of this size. The current study evaluated the effectiveness of this community-directed delivery of doxycycline four years after delivery.

FINDINGS: Infection with O. volvulus was evaluated by skin biopsy and nodule palpation. Of the 507 subjects recruited, 375 had completed the treatment with doxycycline followed by one or two rounds of annual ivermectin MDA and 132 received one or two rounds of annual ivermectin MDA alone. Statistically significant lower microfilarial prevalence (17.0% [doxycycline plus ivermectin group], 27.0% [ivermectin only group], p = 0.014) and load (p = 0.012) were found in people that had received doxycycline followed by ivermectin compared to those who received ivermectin only.

CONCLUSIONS: This study demonstrates the long-term effectiveness of doxycycline treatment delivered with a community-directed strategy even when evaluated four years after delivery in an area of ongoing transmission. This finding shows that a multi-week course of treatment is not a barrier to community-delivery of MDA in restricted populations of this size and supports its implementation to compliment existing control strategies for onchocerciasis, where needed.}, } @article {pmid22427798, year = {2012}, author = {Rasgon, JL}, title = {Wolbachia induces male-specific mortality in the mosquito Culex pipiens (LIN strain).}, journal = {PloS one}, volume = {7}, number = {3}, pages = {e30381}, pmid = {22427798}, issn = {1932-6203}, support = {F31 GM020092/GM/NIGMS NIH HHS/United States ; R21 AI070178/AI/NIAID NIH HHS/United States ; GM020092/GM/NIGMS NIH HHS/United States ; AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Biological Evolution ; Culex/*microbiology/physiology ; Female ; Genetic Fitness/genetics ; Life Tables ; Longevity/*physiology ; Male ; *Mortality ; Sex Factors ; Wolbachia/*pathogenicity/physiology ; }, abstract = {BACKGROUND: Wolbachia are maternally inherited endosymbionts that infect a diverse range of invertebrates, including insects, arachnids, crustaceans and filarial nematodes. Wolbachia are responsible for causing diverse reproductive alterations in their invertebrate hosts that maximize their transmission to the next generation. Evolutionary theory suggests that due to maternal inheritance, Wolbachia should evolve toward mutualism in infected females, but strict maternal inheritance means there is no corresponding force to select for Wolbachia strains that are mutualistic in males.

Using cohort life-table analysis, we demonstrate that in the mosquito Culex pipiens (LIN strain), Wolbachia-infected females show no fitness costs due to infection. However, Wolbachia induces up to a 30% reduction in male lifespan.

CONCLUSIONS/SIGNIFICANCE: These results indicate that the Wolbachia infection of the Culex pipiens LIN strain is virulent in a sex-specific manner. Under laboratory situations where mosquitoes generally mate at young ages, Wolbachia strains that reduce male survival could evolve by drift because increased mortality in older males is not a significant selective force.}, } @article {pmid22419438, year = {2012}, author = {Beukeboom, LW}, title = {Microbial manipulation of host sex determination. Endosymbiotic bacteria can directly manipulate their host's sex determination towards the production of female offspring.}, journal = {BioEssays : news and reviews in molecular, cellular and developmental biology}, volume = {34}, number = {6}, pages = {484-488}, doi = {10.1002/bies.201100192}, pmid = {22419438}, issn = {1521-1878}, mesh = {Animals ; Biological Evolution ; Female ; Feminization/genetics ; Genotype ; Host-Pathogen Interactions ; Lepidoptera/*genetics/microbiology ; Male ; Reproduction/genetics ; Sex Chromosomes/*genetics ; *Sex Determination Processes ; Symbiosis/*genetics ; Wolbachia/metabolism/*pathogenicity ; }, abstract = {A recent study in the lepidopteran Ostrinia scapulalis shows that endosymbionts can actively manipulate the sex determination mechanism of their host. Wolbachia bacteria alter the sex-specific splicing of the doublesex master switch gene. In ZZ males of this female heterogametic system, the female isoform of doublesex is produced in the presence of the bacteria. The effect is a lethal feminization of genotypic males. Curing of ZW females leads to males that die, indicating that the bacteria have an obligate role in proper sex determination and development of their host. Microbial intervention with host sex determination may be a driving force behind the evolutionary turnover of sex determination mechanisms.}, } @article {pmid22411632, year = {2012}, author = {Torres, L and Almazán, C and Ayllón, N and Galindo, RC and Rosario-Cruz, R and Quiroz-Romero, H and Gortazar, C and de la Fuente, J}, title = {Identification of microorganisms in partially fed female horn flies, Haematobia irritans.}, journal = {Parasitology research}, volume = {111}, number = {3}, pages = {1391-1395}, pmid = {22411632}, issn = {1432-1955}, mesh = {Animals ; Cattle/blood ; Diptera/*microbiology/*physiology ; Expressed Sequence Tags ; Female ; Mycobacterium bovis/classification/genetics/*isolation & purification ; Picornaviridae/classification/genetics/*isolation & purification ; RNA-Directed DNA Polymerase ; Real-Time Polymerase Chain Reaction ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {The horn fly Haematobia irritans (Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of cattle. The parasitism of horn flies interferes with cattle feeding, thus reducing weight gain and milk production. Additionally, horn flies are mechanical vectors of pathogens that cause disease in cattle. The aims of this study were to identify microorganisms in partially fed female horn flies through mining of expressed sequence tags (ESTs) and to characterize microorganism prevalence using real-time RT-PCR. Seven unigenes containing 24 ESTs were homologous to infectious agents. Microorganisms identified in partially fed female horn flies ESTs included Nora virus (3 unigenes; 8 ESTs), Wolbachia endosymbionts (3 unigenes; 3 ESTs), and Mycobacterium bovis (1 unigene; 13 ESTs). These results expanded the repertoire of microorganisms that could cause persistent infections or be mechanically transmitted by horn flies and support further studies on the role of horn flies in the epidemiology of these pathogens in Mexico.}, } @article {pmid22408242, year = {2012}, author = {Mavingui, P and Valiente Moro, C and Tran-Van, V and Wisniewski-Dyé, F and Raquin, V and Minard, G and Tran, FH and Voronin, D and Rouy, Z and Bustos, P and Lozano, L and Barbe, V and González, V}, title = {Whole-genome sequence of Wolbachia strain wAlbB, an endosymbiont of tiger mosquito vector Aedes albopictus.}, journal = {Journal of bacteriology}, volume = {194}, number = {7}, pages = {1840}, pmid = {22408242}, issn = {1098-5530}, mesh = {Aedes/*microbiology/physiology ; Animals ; Base Sequence ; *Genome, Bacterial ; Insect Vectors/*microbiology/physiology ; Molecular Sequence Data ; Phylogeny ; Symbiosis ; Wolbachia/classification/*genetics/isolation & purification/physiology ; }, abstract = {Although bacteria of the genus Wolbachia induced significant extended phenotypes to infected hosts, most molecular mechanisms involved are still unknown. To gain insight into the bacterial genetic determinants, we sequenced the whole genome of Wolbachia wAlbB strain, a commensal obligate intracellular of the tiger mosquito Aedes albopictus.}, } @article {pmid22403688, year = {2012}, author = {Wang, L and Zhou, C and He, Z and Wang, ZG and Wang, JL and Wang, YF}, title = {Wolbachia infection decreased the resistance of Drosophila to lead.}, journal = {PloS one}, volume = {7}, number = {3}, pages = {e32643}, pmid = {22403688}, issn = {1932-6203}, mesh = {Animals ; Drosophila melanogaster/*drug effects/immunology/*microbiology/physiology ; *Drug Resistance ; Environmental Pollutants/*toxicity ; Female ; Food Contamination ; Gram-Negative Bacterial Infections/genetics/*immunology/metabolism ; Host-Pathogen Interactions ; Lead/*toxicity ; Longevity/drug effects ; Male ; Malondialdehyde/metabolism ; Oxidative Stress/drug effects ; Superoxide Dismutase/metabolism ; Survival Analysis ; Wolbachia/*pathogenicity/physiology ; }, abstract = {BACKGROUND: The heavy metal lead has been shown to be associated with a genotoxic risk. Drosophila melanogaster is a model organism commonly utilized in genetic toxicology testing. The endosymbionts--Wolbachia are now very common in both wild populations and laboratory stocks of Drosophila. Wolbachia may induce resistance to pathogenic viruses, filarial nematodes and Plasmodium in fruit fly and mosquito hosts. However the effect of Wolbachia infection on the resistance of their hosts to heavy metal is unknown.

Manipulating the lead content in the diet of Drosophila melanogaster, we found that lead consumption had no different effects on developmental time between Wolbachia-infected (Dmel wMel) and -uninfected (Dmel T) flies. While in Pb-contaminated medium, significantly reduced amount of pupae and adults of Dmel wMel were emerged, and Dmel wMel adults had significantly shorter longevity than that of Dmel T flies. Lead infusion in diet resulted in significantly decreased superoxide dismutase (SOD) activity in Dmel T flies (P<0.05), but not in Dmel wMel flies. Correspondingly, lead cultures induced a 10.8 fold increase in malonaldehyde (MDA) contents in Dmel T larvae (P<0.05). While in Dmel wMel larvae, it resulted in only a 1.3 fold increase. By quantitative RT-PCR, we showed that lead infused medium caused significantly increased expression level of relish and CecA2 genes in Dmel T flies (P<0.01). Lead cultures did not change dramatically the expression of these genes in Dmel wMel flies.

CONCLUSIONS/SIGNIFICANCE: These results suggest that Wolbachia infection decreased the resistance of Drosophila to lead likely by limiting the production of peroxides resulted from lead, thus being unable to activate the immunological pathway in the host to prevent them from lead damage. This represents a novel Wolbachia-host interaction and provides information that researchers working on Drosophila toxicology should take in consideration the presence of Wolbachia in the stocks they are analyzing.}, } @article {pmid22392278, year = {2012}, author = {Palayam, M and Lakshminarayanan, K and Radhakrishnan, M and Krishnaswamy, G}, title = {Preliminary analysis to target pyruvate phosphate dikinase from wolbachia endosymbiont of Brugia malayi for designing anti-filarial agents.}, journal = {Interdisciplinary sciences, computational life sciences}, volume = {4}, number = {1}, pages = {74-82}, doi = {10.1007/s12539-011-0109-2}, pmid = {22392278}, issn = {1867-1462}, mesh = {Algorithms ; Amino Acid Sequence ; Amino Acids/metabolism ; Animals ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/*genetics/metabolism ; *Brugia malayi ; DNA, Bacterial ; Databases, Factual ; *Drug Design ; Filariasis/*microbiology ; Genome, Bacterial ; Gluconeogenesis/genetics ; Glycolysis/genetics ; Models, Molecular ; Molecular Sequence Data ; Pyruvate, Orthophosphate Dikinase/*genetics/metabolism ; Sequence Homology ; Signal Transduction/genetics ; Symbiosis ; Wolbachia/enzymology/*genetics ; }, abstract = {Filariasis causing nematode Brugia malayi is shown to harbor wolbachia bacteria as symbionts. The sequenced genome of the wolbachia endosymbiont from B.malayi (wBm) offers an unprecedented opportunity to identify new wolbachia drug targets. Genome analysis of the glycolytic/gluconeogenic pathway has revealed that wBm lacks pyruvate kinase (PK) and may instead utilize the enzyme pyruvate phosphate dikinase (PPDK; ATP: pyruvate, orthophosphate phosphotransferase, EC 2.7.9.1). PPDK catalyses the reversible conversion of AMP, PPi and phosphoenolpyruvate into ATP, Pi and pyruvate. Most organisms including mammals exclusively possess PK. Therefore the absence of PPDK in mammals makes this enzyme as attractive wolbachia drug target. In the present study we have modeled the three dimensional structure of wBm PPDK. The template with 50% identity and 67% similarity in amino acid sequence was employed for homology-modeling approach. The putative active site consists of His476, Arg360, Glu358, Asp344, Arg112, Lys43 and Glu346 was selected as site of interest for designing suitable inhibitor molecules. Docking studies were carried out using induced fit algorithms with OPLS force field of Schrödinger's Glide. The lead molecules which inhibit the PPDK activity are taken from the small molecule library (Pubchem database) and the interaction analysis showed that these compounds may inhibit the function of PPDK in wBm.}, } @article {pmid22386851, year = {2012}, author = {Dhamodharan, R and Hoti, SL and Sankari, T}, title = {Characterization of cofactor-independent phosphoglycerate mutase isoform-1 (Wb-iPGM) gene: a drug and diagnostic target from human lymphatic filarial parasite, Wuchereria bancrofti.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {12}, number = {5}, pages = {957-965}, doi = {10.1016/j.meegid.2012.02.005}, pmid = {22386851}, issn = {1567-7257}, mesh = {Amino Acid Sequence ; Animals ; Base Sequence ; DNA, Complementary/genetics ; Filariasis/*parasitology ; *Genes, Helminth ; Helminth Proteins/chemistry/*genetics/metabolism ; Humans ; Molecular Sequence Annotation ; Molecular Sequence Data ; Phosphoglycerate Mutase/chemistry/*genetics/metabolism ; Phylogeny ; Sequence Alignment ; Sequence Homology, Amino Acid ; Wuchereria bancrofti/*enzymology/*genetics ; }, abstract = {The inter-conversion of 3-phosphoglycerate and 2-phosphoglycerate during glycolysis and gluconeogenesis in filarial nematodes, is catalyzed by a co-factor-independent phosphoglycerate mutase (iPGM). The gene encoding iPGM isoform-1 was amplified from Wuchereria bancrofti, the major causative agent of human lymphatic filariasis. Partial genomic DNA (gDNA) fragment of the gene was also amplified from periodic and sub-periodic forms of W. bancrofti and Brugia malayi and sequenced. The Wb-iPGM isoform-1 gene encodes an ORF of 515 amino acids and is found to share 99.4%, 96.0%, and 64.0% amino acid sequence identity with iPGM of B. malayi, Onchocerca volvulus, and Caenorhabditis elegans, respectively. Serine and all the other 13 amino acid residues involved in the catalytic function of iPGM are highly conserved. Further comparison of iPGM nucleotide and amino acid sequences of Wolbachia of B. malayi with Wb-iPGM showed 41% and 54.4% similarity, respectively. The analysis of partial genomic and amino acid sequences and phylogenetic tree of Wb-iPGM indicated that this gene, apart from being a potential drug target, could provide diagnostic, taxonomical, and evolutionary markers. This is the first report of the characterization of iPGM gene from W. bancrofti.}, } @article {pmid22383972, year = {2012}, author = {Pan, H and Li, X and Ge, D and Wang, S and Wu, Q and Xie, W and Jiao, X and Chu, D and Liu, B and Xu, B and Zhang, Y}, title = {Factors affecting population dynamics of maternally transmitted endosymbionts in Bemisia tabaci.}, journal = {PloS one}, volume = {7}, number = {2}, pages = {e30760}, pmid = {22383972}, issn = {1932-6203}, mesh = {Animals ; Bacteroidetes/*metabolism ; China ; DNA Primers/genetics ; Genetic Variation ; Geography ; Hemiptera/metabolism/*microbiology ; Insecta ; Models, Genetic ; Polymerase Chain Reaction/methods ; Population Dynamics ; Rickettsia/*metabolism ; Species Specificity ; Symbiosis ; Wolbachia/*metabolism ; }, abstract = {While every individual of Bemisia tabaci (Hemiptera: Aleyrodidae) harbors the primary symbiont (P-symbiont) Portiera, the infection frequencies of the six secondary symbionts (S-symbionts) including Hamiltonella, Arsenophonus, Cardinium, Wolbachia, Rickettsia and Fritschea vary greatly among different populations. To characterize the factors influencing the infection dynamics of the six S-symbionts in B. tabaci, gene-specific PCR were conducted to screen for the presence of the P-symbiont Portiera and the six S-symbionts in 61 (17 B and 44 Q biotypes) field populations collected from different plant species and locations in China. All individuals of the 61 populations hosted the P-symbiont Portiera, but none of them harbored Arsenophonus and Fritschea. The presence and infection rates of Hamiltonella, Cardinium, Rickettsia, Wolbachia and their co-infections Rickettsia + Hamiltonella (RH), Rickettsia + Cardinium (RC), Hamiltonella + Cardinium (HC) and Rickettsia + Hamiltonella + Cardinium (RHC) varied significantly among the 61 field populations; and the observed variations can be explained by biotypes, sexes, host plants and geographical locations of these field populations. Taken together, at least three factors including biotype, host plant and geographical location affect the infection dynamics of S-symbionts in B. tabaci.}, } @article {pmid22383881, year = {2012}, author = {Rancès, E and Ye, YH and Woolfit, M and McGraw, EA and O'Neill, SL}, title = {The relative importance of innate immune priming in Wolbachia-mediated dengue interference.}, journal = {PLoS pathogens}, volume = {8}, number = {2}, pages = {e1002548}, pmid = {22383881}, issn = {1553-7374}, mesh = {Aedes/genetics/immunology/microbiology/virology ; Animals ; Antibiosis/genetics/*immunology ; Dengue/genetics/*immunology/microbiology ; Dengue Virus/*immunology/physiology ; Drosophila melanogaster/genetics/immunology/microbiology/virology ; Female ; Gene Expression Profiling ; Immunity, Innate/genetics/*physiology ; Microarray Analysis ; Rickettsiaceae Infections/genetics/immunology/microbiology/virology ; Signal Transduction/genetics/immunology ; Wolbachia/immunology/*physiology ; }, abstract = {The non-virulent Wolbachia strain wMel and the life-shortening strain wMelPop-CLA, both originally from Drosophila melanogaster, have been stably introduced into the mosquito vector of dengue fever, Aedes aegypti. Each of these Wolbachia strains interferes with viral pathogenicity and/or dissemination in both their natural Drosophila host and in their new mosquito host, and it has been suggested that this virus interference may be due to host immune priming by Wolbachia. In order to identify aspects of the mosquito immune response that might underpin virus interference, we used whole-genome microarrays to analyse the transcriptional response of A. aegypti to the wMel and wMelPop-CLA Wolbachia strains. While wMel affected the transcription of far fewer host genes than wMelPop-CLA, both strains activated the expression of some immune genes including anti-microbial peptides, Toll pathway genes and genes involved in melanization. Because the induction of these immune genes might be associated with the very recent introduction of Wolbachia into the mosquito, we also examined the same Wolbachia strains in their original host D. melanogaster. First we demonstrated that when dengue viruses were injected into D. melanogaster, virus accumulation was significantly reduced in the presence of Wolbachia, just as in A. aegypti. Second, when we carried out transcriptional analyses of the same immune genes up-regulated in the new heterologous mosquito host in response to Wolbachia we found no over-expression of these genes in D. melanogaster, infected with either wMel or wMelPop. These results reinforce the idea that the fundamental mechanism involved in viral interference in Drosophila and Aedes is not dependent on the up-regulation of the immune effectors examined, although it cannot be excluded that immune priming in the heterologous mosquito host might enhance the virus interference trait.}, } @article {pmid22380432, year = {2012}, author = {Unckless, RL and Jaenike, J}, title = {Maintenance of a male-killing Wolbachia in Drosophila innubila by male-killing dependent and male-killing independent mechanisms.}, journal = {Evolution; international journal of organic evolution}, volume = {66}, number = {3}, pages = {678-689}, doi = {10.1111/j.1558-5646.2011.01485.x}, pmid = {22380432}, issn = {1558-5646}, mesh = {Animals ; Disease Transmission, Infectious ; Drosophila/genetics/growth & development/*microbiology ; Female ; Fertility ; *Host-Pathogen Interactions ; Inbreeding ; Larva ; Male ; Population Density ; Wings, Animal/growth & development ; Wolbachia/*physiology ; }, abstract = {Many maternally inherited endosymbionts manipulate their host's reproduction in various ways to enhance their own fitness. One such mechanism is male killing (MK), in which sons of infected mothers are killed by the endosymbiont during development. Several hypotheses have been proposed to explain the advantages of MK, including resource reallocation from sons to daughters of infected females, avoidance of inbreeding by infected females, and, if transmission is not purely maternal, the facilitation of horizontal transmission to uninfected females. We tested these hypotheses in Drosophila innubila, a mycophagous species infected with MK Wolbachia. There was no evidence of horizontal transmission in the wild and no evidence Wolbachia reduced levels of inbreeding. Resource reallocation does appear to be operative, as Wolbachia-infected females are slightly larger, on average, than uninfected females, although the selective advantage of larger size is insufficient to account for the frequency of infection in natural populations. Wolbachia-infected females from the wild-although not those from the laboratory-were more fecund than uninfected females. Experimental studies revealed that Wolbachia can boost the fecundity of nutrient-deprived flies and reduce the adverse effect of RNA virus infection. Thus, this MK endosymbiont can provide direct, MK-independent fitness benefits to infected female hosts in addition to possible benefits mediated via MK.}, } @article {pmid22376198, year = {2012}, author = {Machtelinckx, T and Van Leeuwen, T and Van De Wiele, T and Boon, N and De Vos, WH and Sanchez, JA and Nannini, M and Gheysen, G and De Clercq, P}, title = {Microbial community of predatory bugs of the genus Macrolophus (Hemiptera: Miridae).}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S9}, pmid = {22376198}, issn = {1471-2180}, mesh = {Animals ; Female ; Gene Transfer, Horizontal ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Microbiota ; Ovary/microbiology ; Phylogeny ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis/genetics ; Rickettsia/*classification/genetics/*isolation & purification ; Sequence Analysis, RNA/methods ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: The predatory mirids of the genus Macrolophus are key natural enemies of various economically important agricultural pests. Both M. caliginosus and M. pygmaeus are commercially available for the augmentative biological control of arthropod pests in European greenhouses. The latter species is known to be infected with Wolbachia -inducing cytoplasmic incompatibility in its host- but the presence of other endosymbionts has not been demonstrated. In the present study, the microbial diversity was examined in various populations of M. caliginosus and M. pygmaeus by 16S rRNA sequencing and denaturing gradient gel electrophoresis.

RESULTS: Besides Wolbachia, a co-infection of 2 Rickettsia species was detected in all M. pygmaeus populations. Based on a concatenated alignment of the 16S rRNA gene, the gltA gene and the coxA gene, the first is phylogenetically related to Rickettsia bellii, whereas the other is closely related to Rickettsia limoniae. All M. caliginosus populations were infected with the same Wolbachia and limoniae-like Rickettsia strain as M. pygmaeus, but did not harbour the bellii-like Rickettsia strain. Interestingly, individuals with a single infection were not found. A PCR assay on the ovaries of M. pygmaeus and M. caliginosus indicated that all endosymbionts are vertically transmitted. The presence of Wolbachia and Rickettsia in oocytes was confirmed by a fluorescence in situ hybridisation. A bio-assay comparing an infected and an uninfected M. pygmaeus population suggested that the endosymbionts had minor effects on nymphal development of their insect host and did not influence its fecundity.

CONCLUSION: Two species of the palaearctic mirid genus Macrolophus are infected with multiple endosymbionts, including Wolbachia and Rickettsia. Independent of the origin, all tested populations of both M. pygmaeus and M. caliginosus were infected with three and two endosymbionts, respectively. There was no indication that infection with endosymbiotic bacteria had a fitness cost in terms of development and fecundity of the predators.}, } @article {pmid22376177, year = {2012}, author = {Longdon, B and Fabian, DK and Hurst, GD and Jiggins, FM}, title = {Male-killing Wolbachia do not protect Drosophila bifasciata against viral infection.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S8}, pmid = {22376177}, issn = {1471-2180}, support = {WT081279MA//Wellcome Trust/United Kingdom ; //Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Drosophila/*growth & development/*microbiology/physiology/virology ; Female ; Host-Pathogen Interactions ; Insect Viruses/physiology ; Male ; RNA Viruses/*physiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Insect symbionts employ multiple strategies to enhance their spread through populations, and some play a dual role as both a mutualist and a reproductive manipulator. It has recently been found that this is the case for some strains of Wolbachia, which both cause cytoplasmic incompatibility and protect their hosts against viruses. Here, we carry out the first test as to whether a male-killing strain of Wolbachia also provides a direct benefit to its host by providing antiviral protection to its host Drosophila bifasciata. We infected flies with two positive sense RNA viruses known to replicate in a range of Drosophila species (Drosophila C virus and Flock House virus) and measure the rate of death in Wolbachia positive and negative host lines with the same genetic background.

RESULTS: Both viruses caused considerable mortality to D. bifasciata flies, with Drosophila C virus killing 43% more flies than the uninfected controls and Flock House virus killing 78% more flies than the uninfected controls. However, viral induced mortality was unaffected by the presence of Wolbachia.

CONCLUSION: In the first male-killing Wolbachia strain tested for antiviral effects, we found no evidence that it conferred protection against two RNA viruses. We show that although antiviral resistance is widespread across the Wolbachia phylogeny, the trait seems to have been lost or gained along some lineages. We discuss the potential mechanisms of this, and can seemingly discount protection against these viruses as a reason why this symbiont has spread through Drosophila populations.}, } @article {pmid22376153, year = {2012}, author = {Kremer, N and Charif, D and Henri, H and Gavory, F and Wincker, P and Mavingui, P and Vavre, F}, title = {Influence of Wolbachia on host gene expression in an obligatory symbiosis.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S7}, pmid = {22376153}, issn = {1471-2180}, mesh = {Animals ; Female ; Gene Expression Profiling ; *Gene Expression Regulation ; *Genes, Insect ; Host-Pathogen Interactions ; Male ; Ovary/*metabolism/microbiology ; Symbiosis ; Wasps/*genetics/immunology/microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia are intracellular bacteria known to be facultative reproductive parasites of numerous arthropod hosts. Apart from these reproductive manipulations, recent findings indicate that Wolbachia may also modify the host's physiology, notably its immune function. In the parasitoid wasp, Asobara tabida, Wolbachia is necessary for oogenesis completion, and aposymbiotic females are unable to produce viable offspring. The absence of egg production is also associated with an increase in programmed cell death in the ovaries of aposymbiotic females, suggesting that a mechanism that ensures the maintenance of Wolbachia in the wasp could also be responsible for this dependence. In order to decipher the general mechanisms underlying host-Wolbachia interactions and the origin of the dependence, we developed transcriptomic approaches to compare gene expression in symbiotic and aposymbiotic individuals.

RESULTS: As no genetic data were available on A. tabida, we constructed several Expressed Sequence Tags (EST) libraries, and obtained 12,551 unigenes from this species. Gene expression was compared between symbiotic and aposymbiotic ovaries through in silico analysis and in vitro subtraction (SSH). As pleiotropic functions involved in immunity and development could play a major role in the establishment of dependence, the expression of genes involved in oogenesis, programmed cell death (PCD) and immunity (broad sense) was analyzed by quantitative RT-PCR. We showed that Wolbachia might interfere with these numerous biological processes, in particular some related to oxidative stress regulation. We also showed that Wolbachia may interact with immune gene expression to ensure its persistence within the host.

CONCLUSIONS: This study allowed us to constitute the first major dataset of the transcriptome of A. tabida, a species that is a model system for both host/Wolbachia and host/parasitoid interactions. More specifically, our results highlighted that symbiont infection may interfere with numerous pivotal processes at the individual level, suggesting that the impact of Wolbachia should also be investigated beyond reproductive manipulations.}, } @article {pmid22376025, year = {2012}, author = {Doudoumis, V and Tsiamis, G and Wamwiri, F and Brelsfoard, C and Alam, U and Aksoy, E and Dalaperas, S and Abd-Alla, A and Ouma, J and Takac, P and Aksoy, S and Bourtzis, K}, title = {Detection and characterization of Wolbachia infections in laboratory and natural populations of different species of tsetse flies (genus Glossina).}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S3}, pmid = {22376025}, issn = {1471-2180}, support = {AI06892/AI/NIAID NIH HHS/United States ; R03TW008413/TW/FIC NIH HHS/United States ; D43 TW007391/TW/FIC NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; R03 TW008413/TW/FIC NIH HHS/United States ; D43TW007391/TW/FIC NIH HHS/United States ; }, mesh = {Africa ; Animals ; Bacterial Typing Techniques ; Cell Nucleus/genetics ; Gene Transfer, Horizontal ; Genome, Insect ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Tsetse Flies/classification/genetics/*microbiology ; Wolbachia/classification/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia is a genus of endosymbiotic α-Proteobacteria infecting a wide range of arthropods and filarial nematodes. Wolbachia is able to induce reproductive abnormalities such as cytoplasmic incompatibility (CI), thelytokous parthenogenesis, feminization and male killing, thus affecting biology, ecology and evolution of its hosts. The bacterial group has prompted research regarding its potential for the control of agricultural and medical disease vectors, including Glossina spp., which transmits African trypanosomes, the causative agents of sleeping sickness in humans and nagana in animals.

RESULTS: In the present study, we employed a Wolbachia specific 16S rRNA PCR assay to investigate the presence of Wolbachia in six different laboratory stocks as well as in natural populations of nine different Glossina species originating from 10 African countries. Wolbachia was prevalent in Glossina morsitans morsitans, G. morsitans centralis and G. austeni populations. It was also detected in G. brevipalpis, and, for the first time, in G. pallidipes and G. palpalis gambiensis. On the other hand, Wolbachia was not found in G. p. palpalis, G. fuscipes fuscipes and G. tachinoides. Wolbachia infections of different laboratory and natural populations of Glossina species were characterized using 16S rRNA, the wsp (Wolbachia Surface Protein) gene and MLST (Multi Locus Sequence Typing) gene markers. This analysis led to the detection of horizontal gene transfer events, in which Wobachia genes were inserted into the tsetse flies fly nuclear genome.

CONCLUSIONS: Wolbachia infections were detected in both laboratory and natural populations of several different Glossina species. The characterization of these Wolbachia strains promises to lead to a deeper insight in tsetse flies-Wolbachia interactions, which is essential for the development and use of Wolbachia-based biological control methods.}, } @article {pmid22376001, year = {2012}, author = {Jelić, M and Castro, JA and Kurbalija Novičić, Z and Kenig, B and Dimitrijević, D and Savić Veselinović, M and Jovanović, M and Milovanović, D and Stamenković-Radak, M and Andjelković, M}, title = {Absence of linkage disequilibria between chromosomal arrangements and mtDNA haplotypes in natural populations of Drosophila subobscura from the Balkan Peninsula.}, journal = {Genome}, volume = {55}, number = {3}, pages = {214-221}, doi = {10.1139/g2012-004}, pmid = {22376001}, issn = {1480-3321}, mesh = {Animals ; Chromosomes/*genetics ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics ; Haplotypes/*genetics ; *Linkage Disequilibrium ; Restriction Mapping ; Serbia ; Wolbachia/genetics ; }, abstract = {The genetic structure of Drosophila subobscura from the Balkan Peninsula was studied with respect to restriction site polymorphism of mitochondrial DNA in populations from the Derventa River Gorge and Sicevo Gorge (Serbia). To investigate the role of cytonuclear interactions in shaping mitochondrial DNA variability in natural populations of this species, the study was complemented with the analysis of linkage disequilibria between mitochondrial haplotypes and chromosomal inversion arrangements. Similar to other populations of D. subobscura, two main haplotypes (I and II) were found, as well as a series of less common ones. The frequencies of haplotypes I and II accounted for 25.8% and 71.0%, respectively, in the population from the Derventa River Gorge, and for 32.4% and 58.1%, respectively, in the population from Sicevo Gorge. One of the haplotypes harbored a large insertion (2.7 kb) in the A+T rich region. The frequency distribution of both haplotypes did not depart from neutrality. Contrary to prior studies, we did not detect any significant linkage disequilibrium between the two most frequent mtDNA haplotypes and any of the chromosomal arrangements in either of the populations. We conclude that linkage disequilibrium is not a general occurrence in natural populations of D. subobscura, and we discuss how transient coadaptations, ecologically specific selective pressures, and demographics could contribute to population-specific patterns of linkage disequilibrium.}, } @article {pmid22375935, year = {2012}, author = {Zhukova, MV and Kiseleva, E}, title = {The virulent Wolbachia strain wMelPop increases the frequency of apoptosis in the female germline cells of Drosophila melanogaster.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S15}, pmid = {22375935}, issn = {1471-2180}, mesh = {Animals ; Apoptosis ; Drosophila melanogaster/*microbiology/physiology/ultrastructure ; Female ; Germ Cells/microbiology/*physiology/ultrastructure ; Microscopy, Fluorescence ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: Wolbachia are bacterial endosymbionts of many arthropod species in which they manipulate reproductive functions. The distribution of these bacteria in the Drosophila ovarian cells at different stages of oogenesis has been amply described. The pathways along which Wolbachia influences Drosophila oogenesis have been, so far, little studied. It is known that Wolbachia are abundant in the somatic stem cell niche of the Drosophila germarium. A checkpoint, where programmed cell death, or apoptosis, can occur, is located in region 2a/2b of the germarium, which comprises niche cells. Here we address the question whether or not the presence of Wolbachia in germarium cells can affect the frequency of cyst apoptosis in the checkpoint.

RESULTS: Our current fluorescent microscopic observations showed that the wMel and wMelPop strains had different effects on female germline cells of D. melanogaster. The Wolbachia strain wMel did not affect the frequency of apoptosis in cells of the germarium. The presence of the Wolbachia strain wMelPop in the D. melanogasterw1118 ovaries increased the number of germaria where cells underwent apoptosis in the checkpoint. Based on the appearance in the electron microscope, there was no difference in morphological features of apoptotic cystocytes between Wolbachia-infected and uninfected flies. Bacteria with normal ultrastructure and large numbers of degenerating bacteria were found in the dying cyst cells.

CONCLUSIONS: Our current study demonstrated that the Wolbachia strain wMelPop affects the egg chamber formation in the D. melanogaster ovaries. This led to an increase in the number of germaria containing apoptotic cells. It is suggested that Wolbachia can adversely interfere either with the cystocyte differentiation into the oocyte or with the division of somatic stem cells giving rise to follicle cells and, as a consequence, to improper ratio of germline cells to follicle cells and, ultimately, to apoptosis of cysts. There was no similar adverse effect in D. melanogaster Canton S infected with the Wolbachia strain wMel. This was taken to mean that the observed increase in frequency of apoptosis was not the general effect of Wolbachia on germline cells of D. melanogaster, it was rather induced by the virulent Wolbachia strain wMelPop.}, } @article {pmid22375894, year = {2012}, author = {Ros, VI and Fleming, VM and Feil, EJ and Breeuwer, JA}, title = {Diversity and recombination in Wolbachia and Cardinium from Bryobia spider mites.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S13}, pmid = {22375894}, issn = {1471-2180}, mesh = {Animals ; Bacteroides/classification/*genetics ; Genes, Bacterial ; Molecular Sequence Data ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Tetranychidae/classification/*microbiology ; Wolbachia/*classification/*genetics ; }, abstract = {BACKGROUND: Wolbachia and Cardinium are endosymbiotic bacteria infecting many arthropods and manipulating host reproduction. Although these bacteria are maternally transmitted, incongruencies between phylogenies of host and parasite suggest an additional role for occasional horizontal transmission. Consistent with this view is the strong evidence for recombination in Wolbachia, although it is less clear to what extent recombination drives diversification within single host species and genera. Furthermore, little is known concerning the population structures of other insect endosymbionts which co-infect with Wolbachia, such as Cardinium. Here, we explore Wolbachia and Cardinium strain diversity within nine spider mite species (Tetranychidae) from 38 populations, and quantify the contribution of recombination compared to point mutation in generating Wolbachia diversity.

RESULTS: We found a high level of genetic diversity for Wolbachia, with 36 unique strains detected (64 investigated mite individuals). Sequence data from four Wolbachia genes suggest that new alleles are 7.5 to 11 times more likely to be generated by recombination than point mutation. Consistent with previous reports on more diverse host samples, our data did not reveal evidence for co-evolution of Wolbachia with its host. Cardinium was less frequently found in the mites, but also showed a high level of diversity, with eight unique strains detected in 15 individuals on the basis of only two genes. A lack of congruence among host and Cardinium phylogenies was observed.

CONCLUSIONS: We found a high rate of recombination for Wolbachia strains obtained from host species of the spider mite family Tetranychidae, comparable to rates found for horizontally transmitted bacteria. This suggests frequent horizontal transmission of Wolbachia and/or frequent horizontal transfer of single genes. Our findings strengthens earlier reports of recombination for Wolbachia, and shows that high recombination rates are also present on strains from a restrictive host range. Cardinium was found co-infecting several spider mite species, and phylogenetic comparisons suggest also horizontal transmission of Cardinium among hosts.}, } @article {pmid22375862, year = {2012}, author = {Riegler, M and Iturbe-Ormaetxe, I and Woolfit, M and Miller, WJ and O'Neill, SL}, title = {Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S12}, pmid = {22375862}, issn = {1471-2180}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; DNA Fingerprinting/*methods ; DNA, Bacterial/analysis ; Drosophila melanogaster/microbiology ; Genetic Markers ; Genetic Variation ; Genome, Bacterial ; Phylogeny ; Tandem Repeat Sequences ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Strains of the endosymbiotic bacterium Wolbachia pipientis are extremely diverse both genotypically and in terms of their induced phenotypes in invertebrate hosts. Despite extensive molecular characterisation of Wolbachia diversity, little is known about the actual genomic diversity within or between closely related strains that group tightly on the basis of existing gene marker systems, including Multiple Locus Sequence Typing (MLST). There is an urgent need for higher resolution fingerprinting markers of Wolbachia for studies of population genetics, horizontal transmission and experimental evolution.

RESULTS: The genome of the wMel Wolbachia strain that infects Drosophila melanogaster contains inter- and intragenic tandem repeats that may evolve through expansion or contraction. We identified hypervariable regions in wMel, including intergenic Variable Number Tandem Repeats (VNTRs), and genes encoding ankyrin (ANK) repeat domains. We amplified these markers from 14 related Wolbachia strains belonging to supergroup A and were successful in differentiating size polymorphic alleles. Because of their tandemly repeated structure and length polymorphism, the markers can be used in a PCR-diagnostic multilocus typing approach, analogous to the Multiple Locus VNTR Analysis (MLVA) established for many other bacteria and organisms. The isolated markers are highly specific for supergroup A and not informative for other supergroups. However, in silico analysis of completed genomes from other supergroups revealed the presence of tandem repeats that are variable and could therefore be useful for typing target strains.

CONCLUSIONS: Wolbachia genomes contain inter- and intragenic tandem repeats that evolve through expansion or contraction. A selection of polymorphic tandem repeats is a novel and useful PCR diagnostic extension to the existing MLST typing system of Wolbachia, as it allows rapid and inexpensive high-throughput fingerprinting of closely related strains for which polymorphic markers were previously lacking.}, } @article {pmid22375833, year = {2012}, author = {Pinto, SB and Mariconti, M and Bazzocchi, C and Bandi, C and Sinkins, SP}, title = {Wolbachia surface protein induces innate immune responses in mosquito cells.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S11}, pmid = {22375833}, issn = {1471-2180}, support = {079059/WT_/Wellcome Trust/United Kingdom ; 095121/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/immunology/microbiology ; Animals ; Anopheles/*immunology/microbiology ; Bacterial Outer Membrane Proteins/*metabolism ; Cell Line ; Gene Expression Regulation ; Host-Parasite Interactions ; Insect Proteins/*genetics/immunology ; Molecular Sequence Data ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia endosymbiotic bacteria are capable of inducing chronic upregulation of insect immune genes in some situations and this phenotype may influence the transmission of important insect-borne pathogens. However the molecules involved in these interactions have not been characterized.

RESULTS: Here we show that recombinant Wolbachia Surface Protein (WSP) stimulates increased transcription of immune genes in mosquito cells derived from the mosquito Anopheles gambiae, which is naturally uninfected with Wolbachia; at least two of the upregulated genes, TEP1 and APL1, are known to be important in Plasmodium killing in this species. When cells from Aedes albopictus, which is naturally Wolbachia-infected, were challenged with WSP lower levels of upregulation were observed than for the An. gambiae cells.

CONCLUSIONS: We have found that WSP is a strong immune elicitor in a naturally Wolbachia-uninfected mosquito species (Anopheles gambiae) while a milder elicitor in a naturally-infected species (Aedes albopictus). Since the WSP of a mosquito non-native (nematode) Wolbachia strain was used, these data suggest that there is a generalized tolerance to WSP in Ae. albopictus.}, } @article {pmid22375708, year = {2012}, author = {Chevalier, F and Herbinière-Gaboreau, J and Charif, D and Mitta, G and Gavory, F and Wincker, P and Grève, P and Braquart-Varnier, C and Bouchon, D}, title = {Feminizing Wolbachia: a transcriptomics approach with insights on the immune response genes in Armadillidium vulgare.}, journal = {BMC microbiology}, volume = {12 Suppl 1}, number = {Suppl 1}, pages = {S1}, pmid = {22375708}, issn = {1471-2180}, mesh = {Animals ; Expressed Sequence Tags ; Female ; Gene Expression Profiling/*methods ; Gene Expression Regulation ; Isopoda/*genetics/*immunology/microbiology/physiology ; Male ; Ovary/immunology/metabolism ; Symbiosis ; Wolbachia/immunology/*physiology ; }, abstract = {BACKGROUND: Wolbachia are vertically transmitted bacteria known to be the most widespread endosymbiont in arthropods. They induce various alterations of the reproduction of their host, including feminization of genetic males in isopod crustaceans. In the pill bug Armadillidium vulgare, the presence of Wolbachia is also associated with detrimental effects on host fertility and lifespan. Deleterious effects have been demonstrated on hemocyte density, phenoloxidase activity, and natural hemolymph septicemia, suggesting that infected individuals could have defective immune capacities. Since nothing is known about the molecular mechanisms involved in Wolbachia-A. vulgare interactions and its secondary immunocompetence modulation, we developed a transcriptomics strategy and compared A. vulgare gene expression between Wolbachia-infected animals (i.e., "symbiotic" animals) and uninfected ones (i.e., "asymbiotic" animals) as well as between animals challenged or not challenged by a pathogenic bacteria.

RESULTS: Since very little genetic data is available on A. vulgare, we produced several EST libraries and generated a total of 28 606 ESTs. Analyses of these ESTs revealed that immune processes were over-represented in most experimental conditions (responses to a symbiont and to a pathogen). Considering canonical crustacean immune pathways, these genes encode antimicrobial peptides or are involved in pathogen recognition, detoxification, and autophagy. By RT-qPCR, we demonstrated a general trend towards gene under-expression in symbiotic whole animals and ovaries whereas the same gene set tends to be over-expressed in symbiotic immune tissues.

CONCLUSION: This study allowed us to generate the first reference transcriptome ever obtained in the Isopoda group and to identify genes involved in the major known crustacean immune pathways encompassing cellular and humoral responses. Expression of immune-related genes revealed a modulation of host immunity when females are infected by Wolbachia, including in ovaries, the crucial tissue for the Wolbachia route of transmission.}, } @article {pmid22368278, year = {2012}, author = {Weiss, BL and Maltz, M and Aksoy, S}, title = {Obligate symbionts activate immune system development in the tsetse fly.}, journal = {Journal of immunology (Baltimore, Md. : 1950)}, volume = {188}, number = {7}, pages = {3395-3403}, pmid = {22368278}, issn = {1550-6606}, support = {R01 AI081774/AI/NIAID NIH HHS/United States ; AI051584/AI/NIAID NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; 069449//PHS HHS/United States ; R01 AI051584-03/AI/NIAID NIH HHS/United States ; }, mesh = {Adjuvants, Immunologic/pharmacology ; Animals ; Disease Resistance ; Enterobacteriaceae/physiology ; Escherichia coli/pathogenicity ; Female ; Gene Expression Profiling ; Hemocytes/*immunology/transplantation ; Hemolymph/cytology ; Immunity, Cellular ; Immunity, Humoral ; Insect Proteins/biosynthesis/genetics ; Larva/microbiology ; Symbiosis/*immunology ; Tissue Extracts/pharmacology ; Transcription Factors/biosynthesis/genetics ; Tsetse Flies/genetics/growth & development/*immunology/*microbiology ; Wigglesworthia/chemistry/immunology/*physiology ; Wolbachia/physiology ; }, abstract = {Many insects rely on the presence of symbiotic bacteria for proper immune system function. However, the molecular mechanisms that underlie this phenomenon are poorly understood. Adult tsetse flies (Glossina spp.) house three symbiotic bacteria that are vertically transmitted from mother to offspring during this insect's unique viviparous mode of reproduction. Larval tsetse that undergo intrauterine development in the absence of their obligate mutualist, Wigglesworthia, exhibit a compromised immune system during adulthood. In this study, we characterize the immune phenotype of tsetse that develop in the absence of all of their endogenous symbiotic microbes. Aposymbiotic tsetse (Glossina morsitans morsitans [Gmm(Apo)]) present a severely compromised immune system that is characterized by the absence of phagocytic hemocytes and atypical expression of immunity-related genes. Correspondingly, these flies quickly succumb to infection with normally nonpathogenic Escherichia coli. The susceptible phenotype exhibited by Gmm(Apo) adults can be reversed when they receive hemocytes transplanted from wild-type donor flies prior to infection. Furthermore, the process of immune system development can be restored in intrauterine Gmm(Apo) larvae when their mothers are fed a diet supplemented with Wigglesworthia cell extracts. Our finding that molecular components of Wigglesworthia exhibit immunostimulatory activity within tsetse is representative of a novel evolutionary adaptation that steadfastly links an obligate symbiont with its host.}, } @article {pmid22347356, year = {2012}, author = {Evison, SE and Roberts, KE and Laurenson, L and Pietravalle, S and Hui, J and Biesmeijer, JC and Smith, JE and Budge, G and Hughes, WO}, title = {Pervasiveness of parasites in pollinators.}, journal = {PloS one}, volume = {7}, number = {1}, pages = {e30641}, pmid = {22347356}, issn = {1932-6203}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Bees/*parasitology/virology ; DNA, Viral/genetics ; Hymenoptera/*parasitology/virology ; Insect Viruses/genetics/isolation & purification ; Parasites/*pathogenicity ; *Pollination ; Polymerase Chain Reaction ; Wasps/*parasitology/virology ; }, abstract = {Many pollinator populations are declining, with large economic and ecological implications. Parasites are known to be an important factor in the some of the population declines of honey bees and bumblebees, but little is known about the parasites afflicting most other pollinators, or the extent of interspecific transmission or vectoring of parasites. Here we carry out a preliminary screening of pollinators (honey bees, five species of bumblebee, three species of wasp, four species of hoverfly and three genera of other bees) in the UK for parasites. We used molecular methods to screen for six honey bee viruses, Ascosphaera fungi, Microsporidia, and Wolbachia intracellular bacteria. We aimed simply to detect the presence of the parasites, encompassing vectoring as well as actual infections. Many pollinators of all types were positive for Ascosphaera fungi, while Microsporidia were rarer, being most frequently found in bumblebees. We also detected that most pollinators were positive for Wolbachia, most probably indicating infection with this intracellular symbiont, and raising the possibility that it may be an important factor in influencing host sex ratios or fitness in a diversity of pollinators. Importantly, we found that about a third of bumblebees (Bombus pascuorum and Bombus terrestris) and a third of wasps (Vespula vulgaris), as well as all honey bees, were positive for deformed wing virus, but that this virus was not present in other pollinators. Deformed wing virus therefore does not appear to be a general parasite of pollinators, but does interact significantly with at least three species of bumblebee and wasp. Further work is needed to establish the identity of some of the parasites, their spatiotemporal variation, and whether they are infecting the various pollinator species or being vectored. However, these results provide a first insight into the diversity, and potential exchange, of parasites in pollinator communities.}, } @article {pmid22332237, year = {2012}, author = {Wu, M and Scott, AJ}, title = {Phylogenomic analysis of bacterial and archaeal sequences with AMPHORA2.}, journal = {Bioinformatics (Oxford, England)}, volume = {28}, number = {7}, pages = {1033-1034}, doi = {10.1093/bioinformatics/bts079}, pmid = {22332237}, issn = {1367-4811}, mesh = {Algorithms ; Archaea/classification/*genetics ; Bacteria/classification/*genetics ; Computational Biology/*methods ; Genome, Archaeal ; Genome, Bacterial ; Genomics/*methods ; Metagenome ; *Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA/methods ; Sequence Analysis, Protein/methods ; Software ; }, abstract = {SUMMARY: With the explosive growth of bacterial and archaeal sequence data, large-scale phylogenetic analyses present both opportunities and challenges. Here we describe AMPHORA2, an automated phylogenomic inference tool that can be used for high-throughput, high-quality genome tree reconstruction and metagenomic phylotyping. Compared with its predecessor, AMPHORA2 has several major enhancements and new functions: it has a greatly expanded phylogenetic marker database and can analyze both bacterial and archaeal sequences; it incorporates probability-based sequence alignment masks that improve the phylogenetic accuracy; it can analyze DNA as well as protein sequences and is more sensitive in marker identification; finally, it is over 100× faster in metagenomic phylotyping.

AVAILABILITY: http://wolbachia.biology.virginia.edu/WuLab/Software.html.

CONTACT: mw4yv@virginia.edu

SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.}, } @article {pmid22326972, year = {2012}, author = {Kamalakannan, V and Kirthika, S and Haripriya, K and Babu, S and Narayanan, RB}, title = {Wolbachia heat shock protein 60 induces pro-inflammatory cytokines and apoptosis in monocytes in vitro.}, journal = {Microbes and infection}, volume = {14}, number = {7-8}, pages = {610-618}, pmid = {22326972}, issn = {1769-714X}, support = {Y99 AI999999/AI/NIAID NIH HHS/United States ; Z99 AI999999/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Apoptosis/*physiology ; Cell Line ; Chaperonin 60/*metabolism ; Cytokines/*biosynthesis ; Filariasis/immunology/microbiology ; Gene Expression Regulation ; Humans ; Inflammation/*microbiology ; Lymphocytes/immunology ; Monocytes/*immunology ; Nitric Oxide Synthase/genetics ; Recombinant Proteins/metabolism ; Wolbachia/*metabolism ; }, abstract = {Recombinant Wolbachia heat shock protein 60 (rWmhsp60) induces gene expression of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in human monocytic cell line THP-1. In addition, it inhibits the phagocytic activity and does not alter the nitric oxide production by differentiated THP-1 macrophages, which corroborates with no significant change in inducible nitric oxide synthase gene expression in rWmhsp60 treated THP-1 monocytes. Further, 24 h stimulation of peripheral blood mononuclear cells from normal individuals by rWmhsp60 reveals that monocytes enter the late apoptotic stage, while lymphocytes do not show apoptosis. Thus these findings suggest that rWmhsp60 may contribute to inflammation mediated monocyte dysfunction in filarial pathogenesis.}, } @article {pmid22321609, year = {2012}, author = {Rao, RU and Huang, Y and Abubucker, S and Heinz, M and Crosby, SD and Mitreva, M and Weil, GJ}, title = {Effects of doxycycline on gene expression in Wolbachia and Brugia malayi adult female worms in vivo.}, journal = {Journal of biomedical science}, volume = {19}, number = {1}, pages = {21}, pmid = {22321609}, issn = {1423-0127}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/*metabolism ; Doxycycline/*pharmacology ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Bacterial/*drug effects ; Oligonucleotide Array Sequence Analysis ; Wolbachia/*metabolism ; }, abstract = {BACKGROUND: Most filarial nematodes contain Wolbachia symbionts. The purpose of this study was to examine the effects of doxycycline on gene expression in Wolbachia and adult female Brugia malayi.

METHODS: Brugia malayi infected gerbils were treated with doxycycline for 6-weeks. This treatment largely cleared Wolbachia and arrested worm reproduction. RNA recovered from treated and control female worms was labeled by random priming and hybridized to the Version 2- filarial microarray to obtain expression profiles.

RESULTS AND DISCUSSION: Results showed significant changes in expression for 200 Wolbachia (29% of Wolbachia genes with expression signals in untreated worms) and 546 B. malayi array elements after treatment. These elements correspond to known genes and also to novel genes with unknown biological functions. Most differentially expressed Wolbachia genes were down-regulated after treatment (98.5%). In contrast, doxycycline had a mixed effect on B. malayi gene expression with many more genes being significantly up-regulated after treatment (85% of differentially expressed genes). Genes and processes involved in reproduction (gender-regulated genes, collagen, amino acid metabolism, ribosomal processes, and cytoskeleton) were down-regulated after doxycycline while up-regulated genes and pathways suggest adaptations for survival in response to stress (energy metabolism, electron transport, anti-oxidants, nutrient transport, bacterial signaling pathways, and immune evasion).

CONCLUSIONS: Doxycycline reduced Wolbachia and significantly decreased bacterial gene expression. Wolbachia ribosomes are believed to be the primary biological target for doxycycline in filarial worms. B. malayi genes essential for reproduction, growth and development were also down-regulated; these changes are consistent with doxycycline effects on embryo development and reproduction. On the other hand, many B. malayi genes involved in energy production, electron-transport, metabolism, anti-oxidants, and others with unknown functions had increased expression signals after doxycycline treatment. These results suggest that female worms are able to compensate in part for the loss of Wolbachia so that they can survive, albeit without reproductive capacity. This study of doxycycline induced changes in gene expression has provided new clues regarding the symbiotic relationship between Wolbachia and B. malayi.}, } @article {pmid22293464, year = {2012}, author = {Singh, ST and Priya, NG and Kumar, J and Rana, VS and Ellango, R and Joshi, A and Priyadarshini, G and Asokan, R and Rajagopal, R}, title = {Diversity and phylogenetic analysis of endosymbiotic bacteria from field caught Bemisia tabaci from different locations of North India based on 16S rDNA library screening.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {12}, number = {2}, pages = {411-419}, doi = {10.1016/j.meegid.2012.01.015}, pmid = {22293464}, issn = {1567-7257}, mesh = {Animals ; Bacteria/*classification/*genetics ; DNA, Ribosomal/chemistry ; Evolution, Molecular ; *Genetic Variation ; Hemiptera/*microbiology ; India ; *Phylogeny ; Phylogeography ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Bemisia tabaci is the major vector pest of agricultural crops all over the world. In this study we report the different bacterial endosymbionts associated with B. tabaci sampled from 14 different locations in North India. Using 16S rDNA clone library sequences we were able to identify Portiera, the primary endosymbiont of B. tabaci, and other secondary endosymbionts like Cardinium, Wolbachia, Rickettsia and Arsenophonus. Along with these we also detected Bacillus, Enterobacter, Paracoccus and Acinetobacter. These secondary endosymbionts were not uniformly distributed in all the locations. Phylogenetic analysis of 16S rDNA sequences of Cardinium, Wolbachia, Rickettsia and Arsenophonus showed that each of these bacteria form a separate cluster when compared to their respective counterparts from other parts of the world. MtCO1 gene based phylogenetic analysis showed the presence of Asia I and Asia II genetic groups of B. tabaci in N. India. The multiple correspondence analyses showed no correlation between the host genetic group and the endosymbiont diversity. These results suggest that the bacterial endosymbiont diversity of B. tabaci is much larger and complex than previously perceived and probably N. Indian strains of the bacterial symbionts could have evolved from some other ancestor.}, } @article {pmid22292021, year = {2012}, author = {Plantard, O and Bouju-Albert, A and Malard, MA and Hermouet, A and Capron, G and Verheyden, H}, title = {Detection of Wolbachia in the tick Ixodes ricinus is due to the presence of the hymenoptera endoparasitoid Ixodiphagus hookeri.}, journal = {PloS one}, volume = {7}, number = {1}, pages = {e30692}, pmid = {22292021}, issn = {1932-6203}, mesh = {Animals ; Bacterial Outer Membrane Proteins/analysis/genetics ; Female ; Food Chain ; Host-Parasite Interactions/genetics/*physiology ; Host-Pathogen Interactions/genetics/physiology ; Hymenoptera/*microbiology/physiology ; Ixodes/*microbiology/*parasitology/physiology ; Phylogeny ; Symbiosis/physiology ; Tick Infestations/microbiology ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {The identification of micro-organisms carried by ticks is an important issue for human and animal health. In addition to their role as pathogen vectors, ticks are also the hosts for symbiotic bacteria whose impact on tick biology is poorly known. Among these, the bacterium Wolbachia pipientis has already been reported associated with Ixodes ricinus and other tick species. However, the origins of Wolbachia in ticks and their consequences on tick biology (known to be very diverse in invertebrates, ranging from nutritional symbionts in nematodes to reproductive manipulators in insects) are unknown. Here we report that the endoparasitoid wasp Ixodiphagus hookeri (Hymenoptera, Chalcidoidea, Encyrtidae)--strictly associated with ticks for their development--infested at almost 100% prevalence by a W. pipientis strain belonging to a Wolbachia supergroup that has already been reported as associated with other hymenopteran parasitoids. In a natural population of I. ricinus that suffers high parasitism rates due to I. hookeri, we used specific PCR primers for both hymenopteran and W. pipientis gene fragments to show that all unfed tick nymphs parasitized by I. hookeri also harbored Wolbachia, while unparasitized ticks were Wolbachia-free. We demonstrated experimentally that unfed nymphs obtained from larvae exposed to I. hookeri while gorging on their vertebrate host also harbor Wolbachia. We hypothesize that previous studies that have reported W. pipientis in ticks are due to the cryptic presence of the endoparasitoid wasp I. hookeri. This association has remained hidden until now because parasitoids within ticks cannot be detected until engorgement of the nymphs brings the wasp eggs out of diapause. Finally, we discuss the consequences of this finding for our understanding of the tick microbiome, and their possible role in horizontal gene transfer among pathogenic and symbiotic bacteria.}, } @article {pmid22291901, year = {2012}, author = {Riparbelli, MG and Giordano, R and Ueyama, M and Callaini, G}, title = {Wolbachia-mediated male killing is associated with defective chromatin remodeling.}, journal = {PloS one}, volume = {7}, number = {1}, pages = {e30045}, pmid = {22291901}, issn = {1932-6203}, mesh = {Animals ; Chromatin Assembly and Disassembly/genetics/*physiology ; Drosophila/embryology/genetics/*microbiology ; Embryo, Nonmammalian ; Embryonic Development/physiology ; Female ; Genetic Predisposition to Disease ; Male ; Meiosis/genetics/physiology ; Rickettsiaceae Infections/embryology/microbiology/*mortality/veterinary ; *Sex Characteristics ; Spindle Apparatus/genetics/metabolism/physiology ; Wolbachia/*pathogenicity/*physiology ; }, abstract = {Male killing, induced by different bacterial taxa of maternally inherited microorganisms, resulting in highly distorted female-biased sex-ratios, is a common phenomenon among arthropods. Some strains of the endosymbiont bacteria Wolbachia have been shown to induce this phenotype in particular insect hosts. High altitude populations of Drosophila bifasciata infected with Wolbachia show selective male killing during embryonic development. However, since this was first reported, circa 60 years ago, the interaction between Wolbachia and its host has remained unclear. Herein we show that D. bifasciata male embryos display defective chromatin remodeling, improper chromatid segregation and chromosome bridging, as well as abnormal mitotic spindles and gradual loss of their centrosomes. These defects occur at different times in the early development of male embryos leading to death during early nuclear division cycles or large defective areas of the cellular blastoderm, culminating in abnormal embryos that die before eclosion. We propose that Wolbachia affects the development of male embryos by specifically targeting male chromatin remodeling and thus disturbing mitotic spindle assembly and chromosome behavior. These are the first observations that demonstrate fundamental aspects of the cytological mechanism of male killing and represent a solid base for further molecular studies of this phenomenon.}, } @article {pmid22280837, year = {2012}, author = {Tsagkarakou, A and Mouton, L and Kristoffersen, JB and Dokianakis, E and Grispou, M and Bourtzis, K}, title = {Population genetic structure and secondary endosymbionts of Q Bemisia tabaci (Hemiptera: Aleyrodidae) from Greece.}, journal = {Bulletin of entomological research}, volume = {102}, number = {3}, pages = {353-365}, doi = {10.1017/S0007485311000757}, pmid = {22280837}, issn = {1475-2670}, mesh = {Animals ; Ecosystem ; Female ; Gene Flow ; *Genetics, Population ; Greece ; Hemiptera/classification/*genetics/microbiology ; Magnoliopsida/parasitology ; Male ; Microsatellite Repeats ; Phylogeography ; *Polymorphism, Genetic ; *Symbiosis ; Wolbachia/physiology ; }, abstract = {We investigated the molecular diversity of the major agricultural pest Bemisia tabaci and of its associated secondary endosymbionts in Greece. Analyzing mitochondrial DNA, we found that the Q1 (=Q west) is predominant. We used eight microsatellite polymorphic markers to study the genetic structure of 37 populations from mainland and insular Greece, collected on different host species from outdoor and protected crops as well as from non-cultivated plants. In some cases, gene flow was found to be low even between populations separated by just a few kilometres. Bayesian analysis identified two main genetic groups, the first encompassing populations from south Crete and the second composed of populations from north Crete, two other Aegean islands and mainland Greece. Genetic differentiation was not correlated with different host plant species or habitat, or greenhouse versus open environment populations. Gene flow significantly decreased with geographic distance, but no isolation by distance existed when only the samples from mainland Greece or only the samples from Crete were considered. The secondary symbionts Wolbachia and Hamiltonella were present at high frequencies while Arsenophonus, Cardinium and Rickettsia were absent from Greek populations. Multilocus sequence typing of Wolbachia identified two Wolbachia strains. These two strains were found together in most of the populations studied but never in the same host individual. Their role on the observed population structure is discussed.}, } @article {pmid22277830, year = {2012}, author = {Mediannikov, O and Audoly, G and Diatta, G and Trape, JF and Raoult, D}, title = {New Rickettsia sp. in tsetse flies from Senegal.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {35}, number = {2}, pages = {145-150}, doi = {10.1016/j.cimid.2011.12.011}, pmid = {22277830}, issn = {1878-1667}, mesh = {Animals ; Bacterial Proteins/genetics ; Molecular Sequence Data ; Phylogeny ; Rickettsia/classification/*genetics/isolation & purification ; Senegal ; Tsetse Flies/*microbiology ; }, abstract = {Tsetse flies are blood-sucking insects transmitting African trypanosomiasis. They are known to harbor also three intracellular bacteria that play important role in their lifecycle: Wigglesworthia glossinidia, Sodalis glossinidius and Wolbachia sp. We have studied 78 Glossina morsitans submorsitans collected in Senegal. In all studied flies we amplified genes of bacterium phylogenetically close to obligate intracellular pathogen Rickettsia felis, the agent of spotted fever in humans. We also visualized this rickettsia in the cells of tsetse flies by fluorescence in situ hybridization. The role of this probable fourth endosymbiotic bacterium of tsetse flies in Glossina lifecycle and possible pathogenecity for humans should be further investigated.}, } @article {pmid22266203, year = {2012}, author = {Martins, C and Souza, RF and Bueno, OC}, title = {Presence and distribution of the endosymbiont Wolbachia among Solenopsis spp. (Hymenoptera: Formicidae) from Brazil and its evolutionary history.}, journal = {Journal of invertebrate pathology}, volume = {109}, number = {3}, pages = {287-296}, doi = {10.1016/j.jip.2012.01.001}, pmid = {22266203}, issn = {1096-0805}, mesh = {Amino Acid Sequence ; Animals ; Ants/*microbiology ; Bacterial Outer Membrane Proteins/genetics ; *Biological Evolution ; Brazil ; Disease Transmission, Infectious ; Genes, Insect/genetics ; Gram-Negative Bacterial Infections/*epidemiology/*genetics/transmission ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia are intracellular bacteria that commonly infect arthropods. Its prevalence among ants of the genus Solenopsis is high. In the present study, the presence and distribution of these endosymbionts was examined among populations of Solenopsis spp. from Brazil. A phylogenetic analysis based on the wsp gene was conducted to infer the evolutionary history of Wolbachia infections within the populations surveyed. A high frequency of Wolbachia bacteria was observed among the genus Solenopsis, 51% of the colonies examined were infected. Incidence was higher in populations from southern Brazil. However, little genetic variability was found among different Wolbachia strains within supergroups A and B. Our findings also suggest that horizontal transmission events can occur through the social parasite S. daguerrei.}, } @article {pmid22251722, year = {2011}, author = {Olivier-Espejel, S and Sabree, ZL and Noge, K and Becerra, JX}, title = {Gut microbiota in nymph and adults of the giant mesquite bug (Thasus neocalifornicus) (Heteroptera: Coreidae) is dominated by Burkholderia acquired de novo every generation.}, journal = {Environmental entomology}, volume = {40}, number = {5}, pages = {1102-1110}, doi = {10.1603/EN10309}, pmid = {22251722}, issn = {1938-2936}, mesh = {Animals ; Burkholderia/genetics/*isolation & purification ; Culture Techniques ; Female ; Gastrointestinal Tract/microbiology ; Heteroptera/growth & development/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Nymph/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/*genetics ; Sequence Analysis, RNA ; Symbiosis ; }, abstract = {The coreid bug Thasus neocalifornicus Brailovsky and Barrera, commonly known as the giant mesquite bug, is a ubiquitous insect of the southwestern United States. Both nymphs and adults are often found aggregated on mesquite trees (Prosopis spp.: Fabaceae) feeding on seedpods and plant sap. We characterized the indigenous bacterial populations of nymphs and adults of this species by using molecular and phylogenetic techniques and culturing methods. Results show that this insect's bacterial gut community has a limited diversity dominated by Burkholderia associates. Phylogenetic analysis by using 16s rRNA sequences suggests that these β-Proteobacteria are closely related to those symbionts obtained from other heteropteran midgut microbial communities but not to Burkholderia symbionts associated with other insect orders. These bacteria were absent from the eggs and were not found in all younger nymphs, suggesting that they are acquired after the insects have hatched. Rearing experiments of nymphs with potentially Burkholderia contaminated soil suggested that if this symbiont is not acquired, giant mesquite bugs experience higher mortality. Egg, whole-body DNA extractions of younger nymphs, and midgut DNA extractions of fifth-instar nymphs and adults also revealed the presence of α-Proteobacteria from the Wolbachia genus. However, this bacterium was also present in reproductive organs of adults, indicating that this symbiont is not specific to the gut.}, } @article {pmid22251682, year = {2011}, author = {Floate, KD and Coghlin, PC and Dosdall, L}, title = {A test using Wolbachia bacteria to identify Eurasian source populations of cabbage seedpod weevil, Ceutorhynchus obstrictus (Marsham), in North America.}, journal = {Environmental entomology}, volume = {40}, number = {4}, pages = {818-823}, doi = {10.1603/EN10315}, pmid = {22251682}, issn = {1938-2936}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; Europe ; Female ; Genes, Bacterial ; Genes, Mitochondrial ; Introduced Species ; Male ; North America ; Reproduction ; Weevils/*microbiology ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Previous research using mitochondrial haplotypes indicates that North American populations of cabbage seedpod weevil, Ceutorhynchus obstrictus (Marsham), originated from at least two separate introductions from source populations in Eurasia. We tested this hypothesis by comparing the genetic variation of symbiotic Wolbachia bacteria in C. obstrictus among seven North American and four European populations. Because Wolbachia are maternally inherited, infections acquired by a host species at one geographic location theoretically may be present in derivative populations that have established in new regions. Use of the conserved MLST Wolbachia genes gatB, coxA, hcpA, fbpA, and ftsZ identified one strain present in all beetles. Use of the variable wsp gene identified three distinct isolates of this strain that appear to co-occur in all populations and potentially in all individuals. Use of the variable wspB gene provided independent support for the presence of these isolates and evidence of a wspB pseudogene. The lack of genetic variation for these Wolbachia genes among host populations prevents their use to clarify the origins of C. obstrictus in North America. However, the results are an interesting example illustrating disjunction in genetic variation between mitochondria and a maternally-inherited symbiont.}, } @article {pmid22251681, year = {2011}, author = {Gorsane, F and Ben Halima, A and Ben Khalifa, M and Bel-Kadhi, MS and Fakhfakh, H}, title = {Molecular characterization of Bemisia tabaci populations in Tunisia: genetic structure and evidence for multiple acquisition of secondary symbionts.}, journal = {Environmental entomology}, volume = {40}, number = {4}, pages = {809-817}, doi = {10.1603/EN10162}, pmid = {22251681}, issn = {1938-2936}, mesh = {Animals ; Electron Transport Complex IV/genetics ; Genes, Mitochondrial ; *Genetic Variation ; Geography ; Hemiptera/*genetics/microbiology ; Sequence Analysis, DNA ; *Symbiosis ; Tunisia ; }, abstract = {A survey was conducted during 2009-2010 seasons to identify the distribution of Bemisia tabaci (Gennadius) biotypes in Tunisia. The genetic affiliation of collected populations was determined by polymerase chain reaction (PCR)-restriction fragment-length polymorphism (TaqI) of the mitochondrial cytochrom oxidase I (mtCOI) gene. Results, validated by sequencing and phylogenetic analysis, allowed the clustering of sampled sweetpotato whiteflies into B and Q biotypes. As B. tabaci harbors the obligatory bacterium Portiera aleyrodidarum, and a diverse array of secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Cardinium, Arsenophonus, and Fritschea, we report here the infectious status of Tunisian populations by secondary symbionts to find out a correlation between bacterial composition to biotype. The genetic variability and structure of B. tabaci populations in Tunisia was driven by analysis of molecular variance (AMOVA) and the hypothesis of isolation by distance was explored. Selective neutrality and genetic haplotype network tests suggested that Tunisian sweetpotato whiteflies have been undergoing a potential expansion followed by gene flow restriction.}, } @article {pmid22236497, year = {2012}, author = {Albers, A and Esum, ME and Tendongfor, N and Enyong, P and Klarmann, U and Wanji, S and Hoerauf, A and Pfarr, K}, title = {Retarded Onchocerca volvulus L1 to L3 larval development in the Simulium damnosum vector after anti-wolbachial treatment of the human host.}, journal = {Parasites & vectors}, volume = {5}, number = {}, pages = {12}, pmid = {22236497}, issn = {1756-3305}, mesh = {Adult ; Animals ; Anti-Bacterial Agents/*administration & dosage ; Cameroon ; DNA, Bacterial/genetics ; Doxycycline/*administration & dosage ; Human Experimentation ; Humans ; Larva/growth & development/microbiology ; Male ; Middle Aged ; Onchocerca volvulus/*growth & development/microbiology ; Polymerase Chain Reaction ; Simuliidae/*parasitology ; Treatment Outcome ; Wolbachia/*drug effects/isolation & purification ; Young Adult ; }, abstract = {BACKGROUND: The human parasite Onchocerca volvulus harbours Wolbachia endosymbionts essential for worm embryogenesis, larval development and adult survival. In this study, the development of Wolbachia-depleted microfilariae (first stage larvae) to infective third stage larvae (L3) in the insect vector Simulium damnosum was analysed.

METHODS: Infected volunteers in Cameroon were randomly and blindly allocated into doxycycline (200 mg/day for 6 weeks) or placebo treatment groups. After treatment, blackflies were allowed to take a blood meal on the volunteers, captured and dissected for larval counting and DNA extraction for quantitative real-time PCR analysis.

RESULTS: PCR results showed a clear reduction in Wolbachia DNA after doxycycline treatment in microfilariae from human skin biopsies with > 50% reduction at one month post-treatment, eventually reaching a reduction of > 80%. Larval stages recovered from the insect vector had similar levels of reduction of endosymbiotic bacteria. Larval recoveries were analysed longitudinally after treatment to follow the kinetics of larval development. Beginning at three months post-treatment, significantly fewer L3 were seen in the blackflies that had fed on doxycycline treated volunteers. Concomitant with this, the proportion of second stage larvae (L2) was significantly increased in this group.

CONCLUSIONS: Doxycycline treatment and the resulting decline of Wolbachia endobacteria from the microfilaria resulted in retarded development of larvae in the insect vector. Thus, anti-wolbachial treatment could have an additive effect for interrupting transmission by reducing the number of L3 that can be transmitted by blackflies.}, } @article {pmid22233324, year = {2011}, author = {Chai, HN and Du, YZ and Qiu, BL and Zhai, BP}, title = {Detection and phylogenetic analysis of Wolbachia in the Asiatic rice leafroller, Cnaphalocrocis medinalis, in Chinese populations.}, journal = {Journal of insect science (Online)}, volume = {11}, number = {}, pages = {123}, pmid = {22233324}, issn = {1536-2442}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; China ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/genetics ; Moths/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {Wolbachia are a group of intracellular inherited endosymbiontic bacteria infecting a wide range of insects. In this study the infection status of Wolbachia (Rickettsiales: Rickettsiaceae) was measured in the Asiatic rice leafroller, Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae), from twenty locations in China by sequencing wsp, ftsZ and 16S rDNA genes. The results showed high infection rates of Wolbachia in C. medinalis populations. Wolbachia was detected in all geographically separate populations; the average infection rate was ~ 62.5%, and the highest rates were 90% in Wenzhou and Yangzhou populations. The Wolbachia detected in different C. medinalis populations were 100% identical to each other when wsp, ftsZ, and 16S rDNA sequences were compared, with all sequences belonging to the Wolbachia B supergroup. Based on wsp, ftsZ and 16S rDNA sequences of Wolbachia, three phylogenetic trees of similar pattern emerged. This analysis indicated the possibility of inter-species and intra-species horizontal transmission of Wolbachia in different arthropods in related geographical regions. The migration route of C. medinalis in mainland China was also discussed since large differentiation had been found between the wsp sequences of Chinese and Thai populations.}, } @article {pmid22227942, year = {2011}, author = {Ravikumar, H and Prakash, BM and Sampathkumar, S and Puttaraju, HP}, title = {Molecular subgrouping of Wolbachia and bacteriophage WO infection among some Indian Drosophila species.}, journal = {Journal of genetics}, volume = {90}, number = {3}, pages = {507-510}, pmid = {22227942}, issn = {0973-7731}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacteriophages/*genetics/pathogenicity ; Drosophila/genetics/*microbiology/virology ; *Genome, Bacterial ; Genome, Insect ; India ; Phenotype ; Rickettsiaceae Infections/microbiology/transmission ; Wolbachia/classification/genetics/isolation & purification/*pathogenicity/virology ; }, } @article {pmid22219393, year = {2012}, author = {Sugimoto, TN and Ishikawa, Y}, title = {A male-killing Wolbachia carries a feminizing factor and is associated with degradation of the sex-determining system of its host.}, journal = {Biology letters}, volume = {8}, number = {3}, pages = {412-415}, pmid = {22219393}, issn = {1744-957X}, mesh = {Animals ; DNA-Binding Proteins/metabolism ; Female ; Genotype ; Insect Proteins/metabolism ; Japan ; Larva/genetics/growth & development/metabolism/microbiology ; Male ; Moths/genetics/*growth & development/metabolism/*microbiology ; Protein Isoforms/metabolism ; Sex Differentiation ; Sex Ratio ; Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Endosymbiotic bacteria of the genus Wolbachia induce diverse reproductive alterations in their insect hosts. Wolbachia (wSca) infecting the moth Ostrinia scapulalis causes unusual male killing, in which males (genotype: ZZ) selectively die during embryonic and larval development, whereas females (genotype: ZW), in turn, selectively die when cured of infection. To gain insight into the interaction between wSca and the host, we analysed phenotypic and genetic sexes of the embryos and larvae of normal, wSca-infected, and infected-and-cured O. scapulalis by diagnosing the sex-specifically spliced transcripts of Osdsx-a homologue of the sex-determining gene doublesex-and sex chromatin in interphase nuclei, respectively. It was observed that the female-type Osdsx was expressed in the infected male (ZZ) progenies destined to die, whereas the male-type Osdsx was expressed in the cured female (ZW) progenies destined to die. These findings suggest that (i) wSca, a male killer, carries a genetic factor that feminizes the male host, (ii) the sex-determining system of the host is degraded, and (iii) a mismatch between the genetic and phenotypic sexes underlies the sex-specific death.}, } @article {pmid22218612, year = {2012}, author = {Tulgetske, GM and Stouthamer, R}, title = {Characterization of intersex production in Trichogramma kaykai infected with parthenogenesis-inducing Wolbachia.}, journal = {Die Naturwissenschaften}, volume = {99}, number = {2}, pages = {143-152}, pmid = {22218612}, issn = {1432-1904}, mesh = {Animals ; Disorders of Sex Development/microbiology/pathology ; Female ; Male ; Time Factors ; Wasps/anatomy & histology/genetics/*microbiology ; Wolbachia/*physiology ; }, abstract = {Sexually aberrant individuals, displaying both male and female characteristics, are rare in occurrence but are documented throughout the animal kingdom. In parasitoid wasps of the genus Trichogramma, such individuals typically appear as a result of rearing Wolbachia-infected thelytokous wasps at high temperatures. Sexually aberrant Trichogramma have been referred to interchangeably in the literature as gynandromorphs, sexual mosaics and intersexes. However, accurately used, the terms "gynandromorph" and "sexual mosaic" describe an individual composed of a mixture of genetically distinct tissues corresponding to the sexual phenotypes observed, while "intersex" refers to an individual having a uniform genetic constitution but with some tissues exhibiting sexual phenotypes conflicting with the associated genotype. Here, we investigate the heat-induced production of sexually aberrant offspring by thelytokous Trichogramma kaykai. Aberrant individuals were rare, but each was characterized as one of 11 morphotypes ranging from very feminine to very masculine. Overall, the production of aberrant individuals increased with time from the onset of maternal oviposition. However, while the production of males also increased with time, the degree of masculinity of aberrant individuals did not; the different morphotypes appeared to be produced haphazardly. We conclude that the aberrant individuals produced by T. kaykai are actually intersexes and not gynandromorphs. The wasp's close association with Wolbachia and the absence of intersexes in uninfected populations allow us to discuss a possible origin of the condition.}, } @article {pmid22214329, year = {2012}, author = {Genchi, C and Kramer, LH and Sassera, D and Bandi, C}, title = {Wolbachia and its implications for the immunopathology of filariasis.}, journal = {Endocrine, metabolic & immune disorders drug targets}, volume = {12}, number = {1}, pages = {53-56}, doi = {10.2174/187153012799279108}, pmid = {22214329}, issn = {2212-3873}, mesh = {Animals ; Filariasis/*immunology/*microbiology ; Filarioidea/*immunology/*microbiology ; Host-Parasite Interactions/immunology ; Humans ; Immunomodulation/physiology ; Symbiosis/immunology/physiology ; Wolbachia/immunology/*physiology ; }, abstract = {Filarial infections are characterized by immunopathological phenomena, that are responsible for the onset of often dramatic pathological outcomes, such as blindness (Onchocerca volvulus) and elephantiasis (W. bancrofti). In addition, the long-term survival (as long as 10 years) of these parasites in otherwise immunocompetent hosts indicates that these nematodes are capable of manipulating the host immune response. The ground-breaking discovery of the bacterial endosymbiont Wolbachia, which resides in most filarial nematodes causing disease, has led to increasing interest in the role it may play in immuno-modulation, pro-inflammatory pathology and other aspects of filarial infection. Indeed, Wolbachia has been shown to be responsible for exacerbating inflammation (as in river blindness), while at the same time blocking efficient elimination of parasites through the host immune response (Onchocerca ochengi). While studies aimed at identifying Wolbachia as a potential target for anti-filarial therapy are at the forefront of current research, understanding its role in the immunology of filarial infection is a fascinating field that has yet to uncover many secrets.}, } @article {pmid22210220, year = {2012}, author = {Hughes, GL and Vega-Rodriguez, J and Xue, P and Rasgon, JL}, title = {Wolbachia strain wAlbB enhances infection by the rodent malaria parasite Plasmodium berghei in Anopheles gambiae mosquitoes.}, journal = {Applied and environmental microbiology}, volume = {78}, number = {5}, pages = {1491-1495}, pmid = {22210220}, issn = {1098-5336}, support = {R01 AI031478/AI/NIAID NIH HHS/United States ; R21 AI070178/AI/NIAID NIH HHS/United States ; 5R01AI031478/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/parasitology ; Animals ; Anopheles/*parasitology ; Drosophila melanogaster/parasitology ; Gastrointestinal Tract/parasitology ; Mice ; Parasite Egg Count ; Plasmodium berghei/*microbiology/*pathogenicity ; *Symbiosis ; Virulence ; Wolbachia/*physiology ; }, abstract = {Wolbachia, a common bacterial endosymbiont of insects, has been shown to protect its hosts against a wide range of pathogens. However, not all strains exert a protective effect on their host. Here we assess the effects of two divergent Wolbachia strains, wAlbB from Aedes albopictus and wMelPop from Drosophila melanogaster, on the vector competence of Anopheles gambiae challenged with Plasmodium berghei. We show that the wAlbB strain significantly increases P. berghei oocyst levels in the mosquito midgut while wMelPop modestly suppresses oocyst levels. The wAlbB strain is avirulent to mosquitoes while wMelPop is moderately virulent to mosquitoes pre-blood meal and highly virulent after mosquitoes have fed on mice. These various effects on P. berghei levels suggest that Wolbachia strains differ in their interactions with the host and/or pathogen, and these differences could be used to dissect the molecular mechanisms that cause interference of pathogen development in mosquitoes.}, } @article {pmid26467955, year = {2012}, author = {Kageyama, D and Narita, S and Watanabe, M}, title = {Insect Sex Determination Manipulated by Their Endosymbionts: Incidences, Mechanisms and Implications.}, journal = {Insects}, volume = {3}, number = {1}, pages = {161-199}, pmid = {26467955}, issn = {2075-4450}, abstract = {The sex-determining systems of arthropods are surprisingly diverse. Some species have male or female heterogametic sex chromosomes while other species do not have sex chromosomes. Most species are diploids but some species, including wasps, ants, thrips and mites, are haplodiploids (n in males; 2n in females). Many of the sexual aberrations, such as sexual mosaics, sex-specific lethality and conversion of sexuality, can be explained by developmental defects including double fertilization of a binucleate egg, loss of a sex chromosome or perturbation of sex-determining gene expression, which occur accidentally or are induced by certain environmental conditions. However, recent studies have revealed that such sexual aberrations can be caused by various groups of vertically-transmitted endosymbiotic microbes such as bacteria of the genera Wolbachia, Rickettsia, Arsenophonus, Spiroplasma and Cardinium, as well as microsporidian protists. In this review, we first summarize the accumulated data on endosymbiont-induced sexual aberrations, and then discuss how such endosymbionts affect the developmental system of their hosts and what kinds of ecological and evolutionary effects these endosymbionts have on their host populations.}, } @article {pmid22206033, year = {2011}, author = {Atyame, CM and Pasteur, N and Dumas, E and Tortosa, P and Tantely, ML and Pocquet, N and Licciardi, S and Bheecarry, A and Zumbo, B and Weill, M and Duron, O}, title = {Cytoplasmic incompatibility as a means of controlling Culex pipiens quinquefasciatus mosquito in the islands of the south-western Indian Ocean.}, journal = {PLoS neglected tropical diseases}, volume = {5}, number = {12}, pages = {e1440}, pmid = {22206033}, issn = {1935-2735}, mesh = {Animals ; Culex/*growth & development/*microbiology ; Female ; Indian Ocean Islands ; Infertility ; Male ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; Survival Analysis ; Wolbachia/*growth & development ; }, abstract = {The use of the bacterium Wolbachia is an attractive alternative method to control vector populations. In mosquitoes, as in members of the Culex pipiens complex, Wolbachia induces a form of embryonic lethality called cytoplasmic incompatibility, a sperm-egg incompatibility occurring when infected males mate either with uninfected females or with females infected with incompatible Wolbachia strain(s). Here we explore the feasibility of the Incompatible Insect Technique (IIT), a species-specific control approach in which field females are sterilized by inundative releases of incompatible males. We show that the Wolbachia wPip(Is) strain, naturally infecting Cx. p. pipiens mosquitoes from Turkey, is a good candidate to control Cx. p. quinquefasciatus populations on four islands of the south-western Indian Ocean (La Réunion, Mauritius, Grande Glorieuse and Mayotte). The wPip(Is) strain was introduced into the nuclear background of Cx. p. quinquefasciatus mosquitoes from La Réunion, leading to the LR[wPip(Is)] line. Total embryonic lethality was observed in crosses between LR[wPip(Is)] males and all tested field females from the four islands. Interestingly, most crosses involving LR[wPip(Is)] females and field males were also incompatible, which is expected to reduce the impact of any accidental release of LR[wPip(Is)] females. Cage experiments demonstrate that LR[wPip(Is)] males are equally competitive with La Réunion males resulting in demographic crash when LR[wPip(Is)] males were introduced into La Réunion laboratory cages. These results, together with the geographic isolation of the four south-western Indian Ocean islands and their limited land area, support the feasibility of an IIT program using LR[wPip(Is)] males and stimulate the implementation of field tests for a Cx. p. quinquefasciatus control strategy on these islands.}, } @article {pmid22194999, year = {2011}, author = {Bordenstein, SR and Bordenstein, SR}, title = {Temperature affects the tripartite interactions between bacteriophage WO, Wolbachia, and cytoplasmic incompatibility.}, journal = {PloS one}, volume = {6}, number = {12}, pages = {e29106}, pmid = {22194999}, issn = {1932-6203}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/*physiology ; Colony Count, Microbial ; Cytoplasm/*metabolism ; Models, Biological ; Penetrance ; Wasps/*microbiology ; Wolbachia/growth & development/*physiology/*virology ; }, abstract = {Wolbachia infections are a model for understanding intracellular, bacterial symbioses. While the symbiosis is often studied from a binary perspective of host and bacteria, it is increasingly apparent that additional trophic levels can influence the symbiosis. For example, Wolbachia in arthropods harbor a widespread temperate bacteriophage, termed WO, that forms virions and rampantly transfers between coinfections. Here we test the hypothesis that temperatures at the extreme edges of an insect's habitable range alter bacteriophage WO inducibility and in turn, Wolbachia densities and the penetrance of cytoplasmic incompatibility. We report four key findings using the model wasp, Nasonia vitripennis: First, both cold treatment at 18 C and heat treatment at 30 C reduce Wolbachia densities by as much as 74% relative to wasps reared at 25 C. Second, in all cases where Wolbachia densities decline due to temperature changes, phage WO densities increase and inversely associate with Wolbachia densities. Heat has a marked effect on phage WO, yielding phage densities that are 552% higher than the room temperature control. Third, there is a significant affect of insect family on phage WO and endoysmbiont densities. Fourth, at extreme temperatures, there was a temperature-mediated adjustment to the density threshold at which Wolbachia cause complete cytoplasmic incompatibility. Taken together, these results demonstrate that temperature simultaneously affects phage WO densities, endosymbiont densities, and the penetrance of cytoplasmic incompatibility. While temperature shock enhances bacteriophage inducibility and the ensuing bacterial mortality in a wide range of medically and industrially-important bacteria, this is the first investigation of the associations in an obligate intracellular bacteria. Implications to a SOS global sensing feedback mechanism in Wolbachia are discussed.}, } @article {pmid22193955, year = {2011}, author = {Serbus, LR and Ferreccio, A and Zhukova, M and McMorris, CL and Kiseleva, E and Sullivan, W}, title = {A feedback loop between Wolbachia and the Drosophila gurken mRNP complex influences Wolbachia titer.}, journal = {Journal of cell science}, volume = {124}, number = {Pt 24}, pages = {4299-4308}, pmid = {22193955}, issn = {1477-9137}, support = {F32 GM080192/GM/NIGMS NIH HHS/United States ; R25 GM058903/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila Proteins/analysis/*genetics ; Drosophila melanogaster/genetics/microbiology/ultrastructure ; Feedback, Physiological ; Microtubules/physiology ; Oocytes/microbiology/ultrastructure ; Oogenesis ; RNA-Binding Proteins/analysis ; Ribonucleoproteins/*physiology ; Symbiosis ; Transforming Growth Factor alpha/*genetics ; Wolbachia/*physiology/ultrastructure ; }, abstract = {Although much is known about interactions between bacterial endosymbionts and their hosts, little is known concerning the host factors that influence endosymbiont titer. Wolbachia endosymbionts are globally dispersed throughout most insect species and are the causative agent in filarial nematode-mediated disease. Our investigation indicates that gurken (grk), a host gene encoding a crucial axis determinant, has a cumulative, dosage-sensitive impact on Wolbachia growth and proliferation during Drosophila oogenesis. This effect appears to be mediated by grk mRNA and its protein-binding partners Squid and Hrp48/Hrb27C, implicating the grk mRNA-protein (mRNP) complex as a rate-limiting host factor controlling Wolbachia titer. Furthermore, highly infected flies exhibit defects that match those occurring with disruption of grk mRNPs, such as nurse cell chromatin disruptions and malformation of chorionic appendages. These findings suggest a feedback loop in which Wolbachia interaction with the grk mRNP affects both Wolbachia titer and grk mRNP function.}, } @article {pmid22192469, year = {2012}, author = {Schraiber, JG and Kaczmarczyk, AN and Kwok, R and Park, M and Silverstein, R and Rutaganira, FU and Aggarwal, T and Schwemmer, MA and Hom, CL and Grosberg, RK and Schreiber, SJ}, title = {Constraints on the use of lifespan-shortening Wolbachia to control dengue fever.}, journal = {Journal of theoretical biology}, volume = {297}, number = {}, pages = {26-32}, doi = {10.1016/j.jtbi.2011.12.006}, pmid = {22192469}, issn = {1095-8541}, mesh = {Aedes/*microbiology/physiology/virology ; Animals ; Dengue/*prevention & control/transmission ; Fertility/physiology ; Host-Pathogen Interactions ; Humans ; Insect Vectors/*microbiology/virology ; Longevity ; Models, Biological ; Pest Control, Biological/*methods ; Population Dynamics ; Wolbachia/*physiology ; }, abstract = {Dengue fever, a viral disease spread by the mosquito Aedes aegypti, affects 50-100 million people a year in many tropical countries. Because the virus must incubate within mosquito hosts for two weeks before being able to transmit the infection, shortening the lifespan of mosquitoes may curtail dengue transmission. We developed a continuous time reaction-diffusion model of the spatial spread of Wolbachia through a population of A. aegypti. This model incorporates the lifespan-shortening effects of Wolbachia on infected A. aegypti and the fitness advantage to infected females due to cytoplasmic incompatibility (CI). We found that local establishment of the Wolbachia infection can occur if the fitness advantage due to CI exceeds the fitness reduction due to lifespan-shortening effects, in accordance with earlier results concerning fecundity reduction. However, spatial spread is possible only if the fitness advantage due to CI is twice as great as the fitness reduction due to lifespan shortening effects. Moreover, lifespan-shortening and fecundity-reduction can have different effects on the speed of wave-retreat. Using data from the literature, we estimated all demographic parameters for infected and uninfected mosquitoes and computed the velocities of spread of infection. Our most optimistic estimates suggest that the spatial spread of lifespan-shortening Wolbachia may be so slow that efficient spatial spread would require a prohibitively large number of point releases. However, as these estimates of demographic parameters may not accurately reflect natural conditions, further research is necessary to corroborate these predictions.}, } @article {pmid22188300, year = {2012}, author = {Verne, S and Johnson, M and Bouchon, D and Grandjean, F}, title = {Effects of parasitic sex-ratio distorters on host genetic structure in the Armadillidium vulgare-Wolbachia association.}, journal = {Journal of evolutionary biology}, volume = {25}, number = {2}, pages = {264-276}, doi = {10.1111/j.1420-9101.2011.02413.x}, pmid = {22188300}, issn = {1420-9101}, mesh = {Animals ; DNA, Mitochondrial/chemistry ; Female ; Humans ; Isopoda/genetics/*microbiology ; Linkage Disequilibrium ; Male ; Microsatellite Repeats ; Polymorphism, Genetic ; Sex Determination Processes/genetics ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {In the pill bug Armadillidium vulgare (Crustacea, Oniscidea), Wolbachia facilitates its spread through vertical transmission via the eggs by inducing feminization of genetic males. The spread of feminizing Wolbachia within and across populations is therefore expected to influence mitochondrial DNA (mtDNA) genetic structure by hitchhiking. To test this hypothesis, we analysed nuclear and mtDNA genetic structure, and Wolbachia prevalence in 13 populations of the pill bug host. Wolbachia prevalence (ranging from 0% to 100% of sampled females) was highly variable among populations. All three Wolbachia strains previously observed in A. vulgare were present (wVulC, wVulM and wVulP) with wVulC being the most prevalent (nine of 13 populations). The host showed a genetic structure on five microsatellite loci that is compatible with isolation by distance. The strong genetic structure observed on host mtDNA was correlated with Wolbachia prevalence: three mitotypes were in strong linkage disequilibrium with the three strains of Wolbachia. Neutrality tests showed that the mtDNA polymorphism is not neutral, and we thus suggest that this unusual pattern of mtDNA polymorphism found in A. vulgare was due to Wolbachia.}, } @article {pmid22182607, year = {2011}, author = {Hail, D and Lauzìere, I and Dowd, SE and Bextine, B}, title = {Culture independent survey of the microbiota of the glassy-winged sharpshooter (Homalodisca vitripennis) using 454 pyrosequencing.}, journal = {Environmental entomology}, volume = {40}, number = {1}, pages = {23-29}, doi = {10.1603/EN10115}, pmid = {22182607}, issn = {1938-2936}, mesh = {Animals ; Bacteria/*classification/genetics ; Bacterial Typing Techniques ; Biodiversity ; DNA, Bacterial/genetics ; Gastrointestinal Tract/microbiology ; Hemiptera/*microbiology ; Hemolymph/microbiology ; *Metagenome ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Symbiosis ; }, abstract = {The glassy-winged sharpshooter, Homalodisca vitripennis (Germar), is an invasive pest that has spread across the southern and western United States. H. vitripennis is highly polyphagous and voracious, feeding on at least 100 plant species and consuming up to 100 times its weight in xylem fluid daily. The insect is a vector of the phytopathogen Xylella fastidiosa (Wells), which is the causative agent of Pierce's disease in grapevines. To evaluate the microbial flora associated with H. vitripennis, total DNA extracts from hemolymph, alimentary canal excretions, and whole insect bodies were subjected to 16S rDNA pyrosequencing using the bTEFAP methodology and the resulting sequences (370-520 bp in length) were compared with a curated high quality 16S database derived from GenBank http://www.ncbi.nlm.nih.gov. Species from the genera Wolbachia, Delftia (formerly Pseudomonas), Pectobacterium, Moraxella, Serratia, Bacillus, and many others were detected and a comprehensive picture of the microbiome associated with H. vitripennis was established. Some of the bacteria identified in this report are initial discoveries; providing a breadth of knowledge to the microbial flora of this insect pest can serve as a reservoir of information for developing biological control strategies.}, } @article {pmid22182545, year = {2010}, author = {Zhang, H and Zhang, KJ and Hong, XY}, title = {Population dynamics of noncytoplasmic incompatibility-inducing Wolbachia in Nilaparvata lugens and its effects on host adult life span and female fitness.}, journal = {Environmental entomology}, volume = {39}, number = {6}, pages = {1801-1809}, doi = {10.1603/EN10051}, pmid = {22182545}, issn = {1938-2936}, mesh = {Animals ; China ; Female ; Fertility ; Hemiptera/*microbiology/physiology ; *Longevity ; Male ; Nymph/microbiology ; *Oviparity ; Wolbachia/*physiology ; }, abstract = {Wolbachia are bacteria that live intracellularly in a wide variety of arthropods. They are maternally inherited and can affect both reproduction and fitness of its host. When infected males mate with uninfected females or females infected by a different Wolbachia strain, there is often a failure of karyogamy, which is usually attributed to cytoplasmic incompatibility (CI). We measured the strength of CI induced by Wolbachia and the fitness effects in three Chinese populations of the brown planthopper Nilaparvata lugens from Hainan, Yunnan, and Guangxi provinces, respectively. No evidence for CI was found in any of the populations, whereas an enhanced fecundity and shortened longevity were observed only in the Hainan population. The infection density was significantly higher in the Hainan population than in the Guangxi population. The Wolbachia strain infecting the three populations appeared to be the same based on the nucleotide sequence of the wsp gene. Therefore, the variable effects of Wolbachia on host fitness seem to be the result of differences in the host genetic background and Wolbachia infection density. The ability of the non-CI-inducing Wolbachia to maintain themselves in their hosts may be attributed to their positive effects on host fecundity and efficient maternal transmission.}, } @article {pmid22174869, year = {2011}, author = {Augustinos, AA and Santos-Garcia, D and Dionyssopoulou, E and Moreira, M and Papapanagiotou, A and Scarvelakis, M and Doudoumis, V and Ramos, S and Aguiar, AF and Borges, PA and Khadem, M and Latorre, A and Tsiamis, G and Bourtzis, K}, title = {Detection and characterization of Wolbachia infections in natural populations of aphids: is the hidden diversity fully unraveled?.}, journal = {PloS one}, volume = {6}, number = {12}, pages = {e28695}, pmid = {22174869}, issn = {1932-6203}, mesh = {Alleles ; Animals ; Aphids/*microbiology ; DNA, Ribosomal/genetics ; *Genetic Variation ; Genotype ; Gram-Negative Bacterial Infections/*microbiology ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny ; Polymerase Chain Reaction ; Population Dynamics ; RNA, Ribosomal, 16S/genetics ; Wolbachia/*genetics/*isolation & purification ; }, abstract = {Aphids are a serious threat to agriculture, despite being a rather small group of insects. The about 4,000 species worldwide engage in highly interesting and complex relationships with their microbial fauna. One of the key symbionts in arthropods is Wolbachia, an α-Proteobacterium implicated in many important biological processes and believed to be a potential tool for biological control. Aphids were thought not to harbour Wolbachia; however, current data suggest that its presence in aphids has been missed, probably due to the low titre of the infection and/or to the high divergence of the Wolbachia strains of aphids. The goal of the present study is to map the Wolbachia infection status of natural aphids populations, along with the characterization of the detected Wolbachia strains. Out of 425 samples from Spain, Portugal, Greece, Israel and Iran, 37 were found to be infected. Our results, based mainly on 16S rRNA gene sequencing, indicate the presence of two new Wolbachia supergroups prevailing in aphids, along with some strains belonging either to supergroup B or to supergroup A.}, } @article {pmid22174680, year = {2011}, author = {Alam, U and Medlock, J and Brelsfoard, C and Pais, R and Lohs, C and Balmand, S and Carnogursky, J and Heddi, A and Takac, P and Galvani, A and Aksoy, S}, title = {Wolbachia symbiont infections induce strong cytoplasmic incompatibility in the tsetse fly Glossina morsitans.}, journal = {PLoS pathogens}, volume = {7}, number = {12}, pages = {e1002415}, pmid = {22174680}, issn = {1553-7374}, support = {R01 AI068932/AI/NIAID NIH HHS/United States ; R01 GM069449/GM/NIGMS NIH HHS/United States ; AI06892/AI/NIAID NIH HHS/United States ; GM069449/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm ; Disease Resistance/*physiology ; Female ; Fertility/genetics ; In Situ Hybridization, Fluorescence ; Insect Vectors/genetics ; Male ; *Models, Theoretical ; Pest Control, Biological/*methods ; Phenotype ; Symbiosis/genetics ; Tsetse Flies/genetics/*microbiology ; *Wolbachia ; }, abstract = {Tsetse flies are vectors of the protozoan parasite African trypanosomes, which cause sleeping sickness disease in humans and nagana in livestock. Although there are no effective vaccines and efficacious drugs against this parasite, vector reduction methods have been successful in curbing the disease, especially for nagana. Potential vector control methods that do not involve use of chemicals is a genetic modification approach where flies engineered to be parasite resistant are allowed to replace their susceptible natural counterparts, and Sterile Insect technique (SIT) where males sterilized by chemical means are released to suppress female fecundity. The success of genetic modification approaches requires identification of strong drive systems to spread the desirable traits and the efficacy of SIT can be enhanced by identification of natural mating incompatibility. One such drive mechanism results from the cytoplasmic incompatibility (CI) phenomenon induced by the symbiont Wolbachia. CI can also be used to induce natural mating incompatibility between release males and natural populations. Although Wolbachia infections have been reported in tsetse, it has been a challenge to understand their functional biology as attempts to cure tsetse of Wolbachia infections by antibiotic treatment damages the obligate mutualistic symbiont (Wigglesworthia), without which the flies are sterile. Here, we developed aposymbiotic (symbiont-free) and fertile tsetse lines by dietary provisioning of tetracycline supplemented blood meals with yeast extract, which rescues Wigglesworthia-induced sterility. Our results reveal that Wolbachia infections confer strong CI during embryogenesis in Wolbachia-free (Gmm(Apo)) females when mated with Wolbachia-infected (Gmm(Wt)) males. These results are the first demonstration of the biological significance of Wolbachia infections in tsetse. Furthermore, when incorporated into a mathematical model, our results confirm that Wolbachia can be used successfully as a gene driver. This lays the foundation for new disease control methods including a population replacement approach with parasite resistant flies. Alternatively, the availability of males that are reproductively incompatible with natural populations can enhance the efficacy of the ongoing sterile insect technique (SIT) applications by eliminating the need for chemical irradiation.}, } @article {pmid22172602, year = {2012}, author = {Martin, OY and Gubler, A and Wimmer, C and Germann, C and Bernasconi, MV}, title = {Infections with Wolbachia and Spiroplasma in the Scathophagidae and other Muscoidea.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {12}, number = {2}, pages = {315-323}, doi = {10.1016/j.meegid.2011.11.012}, pmid = {22172602}, issn = {1567-7257}, mesh = {Animals ; DNA, Bacterial ; Diptera/*microbiology ; Molecular Sequence Data ; Phylogeny ; Spiroplasma/classification/*genetics ; Wolbachia/classification/*genetics ; }, abstract = {The microbes Wolbachia and Spiroplasma are common reproductive parasites of arthropods and may strongly influence reproduction of infected hosts and also impact on reproductive isolation. Such infections could hence influence results of many studies assessing reproductive behaviour and fitness of possible hosts, as well as reproductive isolation. Previous work indicates that infections with the microbes Wolbachia and Spiroplasma are common in the Drosophilidae. However, extensive and targeted surveys of other Dipteran families are lacking. Here we survey the yellow dung fly Scathophaga stercoraria and a range of other species from the Muscoidea (families Scathophagidae, Anthomyiidae, Fanniidae and Muscidae) collected in the field or obtained from museum collections for infection with the widespread reproductive parasites Wolbachia and Spiroplasma. Both have been shown to be heritable symbionts and affect reproduction in other Diptera. S. stercoraria is a very important model for the study of sexual selection, and in particular of postcopulatory processes, as it has played a major role in the history of research on sperm competition and cryptic female choice. Infections with Wolbachia were found to be widespread across the Muscoidea, whereas infections with Spiroplasma were rarer. We discuss the consequences of these findings and directions for future research on the impact of reproductive parasites on host reproduction in the Scathophagidae.}, } @article {pmid22152674, year = {2011}, author = {Tijsse-Klasen, E and Braks, M and Scholte, EJ and Sprong, H}, title = {Parasites of vectors--Ixodiphagus hookeri and its Wolbachia symbionts in ticks in The Netherlands.}, journal = {Parasites & vectors}, volume = {4}, number = {}, pages = {228}, pmid = {22152674}, issn = {1756-3305}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Genes, rRNA ; Hymenoptera/genetics/growth & development/*microbiology ; Ixodes/parasitology ; Netherlands ; RNA, Ribosomal, 28S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*isolation & purification/*physiology ; }, abstract = {BACKGROUND: Ixodiphagus hookeri is a parasitic wasp of ixodid ticks around the world. It has been studied as a potential bio-control agent for several tick species. We suspected that the presence of Wolbachia infected I. hookeri eggs in ticks is responsible for incidental detection of Wolbachia DNA in tick samples.

METHODS: The 28S rRNA and 16S rRNA genes of a specimen of I. hookeri was amplified and sequenced. PCR on part of the 28S rRNA gene was used to detect parasitic wasp DNA in 349 questing Ixodes ricinus ticks from various sampling sites. Furthermore, the wsp gene of Wolbachia was sequenced from the I. hookeri specimen and a subset of ticks was tested using this marker.

RESULTS: Several sequences from tick specimens were identical to the Wolbachia sequence of the I. hookeri specimen. Ixodiphagus hookeri was detected in 9.5% of all tested ticks, varying between 4% and 26% depending on geographic location. Ten out of eleven sampling sites throughout the Netherlands were positive for I. hookeri. Eighty-seven percent of I. hookeri-positive but only 1.6% of I. hookeri-negative ticks were Wolbachia positive. Detection of I. hookeri DNA was strongly associated with the detection of Wolbachia in ticks.

CONCLUSION: This is the first reported case of I. hookeri in the Netherlands. Furthermore I. hookeri harbours Wolbachia species and is broadly distributed in the Netherlands. While detection of Wolbachia DNA in ticks might often be due to parasitism with this wasp, other sources of Wolbachia DNA in ticks might exist as well.}, } @article {pmid22152604, year = {2011}, author = {Bowman, DD}, title = {Introduction to the alpha-proteobacteria: Wolbachia and Bartonella, Rickettsia, Brucella, Ehrlichia, and Anaplasma.}, journal = {Topics in companion animal medicine}, volume = {26}, number = {4}, pages = {173-177}, doi = {10.1053/j.tcam.2011.09.002}, pmid = {22152604}, issn = {1946-9837}, mesh = {Anaplasma/growth & development ; Animals ; Bartonella/growth & development ; Brucella/growth & development ; Cat Diseases/drug therapy/parasitology ; Cats ; Dirofilaria immitis/drug effects/*microbiology ; Dirofilariasis/*drug therapy/parasitology ; Dog Diseases/drug therapy/parasitology ; Dogs ; Doxycycline/*therapeutic use ; Ehrlichia/growth & development ; Filaricides/*therapeutic use ; Rickettsia/growth & development ; Symbiosis ; Wolbachia/growth & development ; }, abstract = {Wolbachia is an obligate intracellular endosymbiont and likely mutualist living within the heartworm Dirofilaria immitis and a number of other filarial nematodes in the family Onchocercidae. The bacterial infection is passed from worm to worm transovarially; the organisms are in ovarian cells, the developing microfilariae, and multiply and persist in all later developmental stages through the mosquito and into the next host. Besides being present in the ovaries of the adult worms, they also are present in large numbers within the hypodermal tissues of the nematode. It is now know that these bacteria that were first observed in heartworms more than 30 years ago are actually related to similar Wolbachia bacteria that are found in arthropods. Wolbachia is an alpha-proteobacteria, and this group includes a number of important arthropod-transmitted bacterial agents of dogs and cats: Rickettsia rickettsii, R. felis, Anaplasma platys, Ehrlichia canis, E. chaffeensis, and E. ewingii. Alpha-proteobacteria are also important as obligate intracellular mutualists in plants in which they are responsible for nitrogen fixation. Recent work on the treatment of heartworms in dogs with doxycycline stems from related work with the human filarial nematode Onchocerca volvulus that causes river blindness in people.}, } @article {pmid22151691, year = {2011}, author = {Muñoz, AG and Baxter, SW and Linares, M and Jiggins, CD}, title = {Deep mitochondrial divergence within a Heliconius butterfly species is not explained by cryptic speciation or endosymbiotic bacteria.}, journal = {BMC evolutionary biology}, volume = {11}, number = {}, pages = {358}, pmid = {22151691}, issn = {1471-2148}, mesh = {Amplified Fragment Length Polymorphism Analysis ; Animals ; Bacteria ; Butterflies/classification/*genetics/microbiology ; Cluster Analysis ; Colombia ; DNA, Mitochondrial/*genetics ; *Evolution, Molecular ; *Genetic Speciation ; Genetics, Population ; Haplotypes ; Phylogeny ; Polymorphism, Genetic ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {BACKGROUND: Cryptic population structure can be an indicator of incipient speciation or historical processes. We investigated a previously documented deep break in the mitochondrial haplotypes of Heliconius erato chestertonii to explore the possibility of cryptic speciation, and also the possible presence of endosymbiont bacteria that might drive mitochondrial population structure.

RESULTS: Among a sample of 315 individuals from 16 populations of western Colombia, two principal mtDNA clades were detected with 2.15% divergence and we confirmed this structure was weakly associated with geography. The first mtDNA clade included 87% of individuals from northern populations and was the sister group of H. erato members of Andes western, while the second clade contained most individuals from southern populations (78%), which shared haplotypes with an Ecuadorian race of H. erato. In contrast, analysis using AFLP markers showed H. e. chestertonii to be a genetically homogeneous species with no association between mitochondrial divergence and AFLP structure. The lack of congruence between molecular markers suggests that cryptic speciation is not a plausible explanation for the deep mitochondrial divergence in H. e chestertonii. We also carried out the first tests for the presence of endosymbiontic bacteria in Heliconius, and identified two distinct lineages of Wolbachia within H. e. chestertonii. However, neither of the principal mitochondrial clades of H. e. chestertonii was directly associated with the patterns of infection.

CONCLUSIONS: We conclude that historical demographic processes are the most likely explanation for the high mitochondrial differentiation in H. e. chestertonii, perhaps due to gene flow between Cauca valley H. e. chestertonii and west Pacific slope populations of H. erato.}, } @article {pmid22145623, year = {2011}, author = {Zheng, Y and Wang, JL and Liu, C and Wang, CP and Walker, T and Wang, YF}, title = {Differentially expressed profiles in the larval testes of Wolbachia infected and uninfected Drosophila.}, journal = {BMC genomics}, volume = {12}, number = {}, pages = {595}, pmid = {22145623}, issn = {1471-2164}, mesh = {Animals ; Drosophila melanogaster ; *Gene Expression Profiling ; In Situ Hybridization ; Larva/*metabolism ; Male ; Polymerase Chain Reaction ; Testis/growth & development/*metabolism/microbiology ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Wolbachia are endosymbiotic bacteria that are frequently found in arthropods and nematodes. These maternally inherited bacteria manipulate host reproduction by several mechanisms including cytoplasmic incompatibility (CI). CI is the most common phenotype induced by Wolbachia and results in the developmental arrest of embryos derived from crosses between Wolbachia-infected males and uninfected females. Although the molecular mechanisms of CI are currently unknown, several studies suggest that host sperm is modified by Wolbachia during spermatogenesis.

RESULTS: We compared the gene expression of Drosophila melanogaster larval testes with and without the wMel strain of Wolbachia to identify candidate genes that could be involved in the interaction between Wolbachia and the insect host. Microarray, quantitative RT-PCR and in situ hybridization analyses were carried out on D. melanogaster larval testes to determine the effect of Wolbachia infection on host gene expression. A total of 296 genes were identified by microarray analysis to have at least a 1.5 fold change [q-value < 5%] in expression. When comparing Wolbachia-infected flies to uninfected flies, 167 genes were up-regulated and 129 genes down-regulated. Differential expression of genes related to metabolism, immunity, reproduction and other functions were observed. Quantitative RT-PCR (qRT-PCR) confirmed 12 genes are differentially expressed in the testes of the 3rd instar larvae of Wolbachia-infected and uninfected flies. In situ hybridization demonstrated that Wolbachia infection changes the expression of several genes putatively associated with spermatogenesis including JH induced protein-26 and Mst84Db, or involved in immune (kenny) or metabolism (CG4988-RA).

CONCLUSIONS: Wolbachia change the gene expression of 296 genes in the larval testes of D. melanogaster including genes related to metabolism, immunity and reproduction. Interestingly, most of the genes putatively involved in immunity were up-regulated in the presence of Wolbachia. In contrast, most of the genes putatively associated with reproduction (especially spermatogenesis) were down-regulated in the presence of Wolbachia. These results suggest Wolbachia may activate the immune pathway but inhibit spermatogenesis. Our data provide a significant panel of candidate genes that may be involved in the interaction between Wolbachia and their insect hosts. This forms a basis to help elucidate the underlying mechanisms of Wolbachia-induced CI in Drosophila and the influence of Wolbachia on spermatogenesis.}, } @article {pmid22140592, year = {2011}, author = {Li, Z and Garner, AL and Gloeckner, C and Janda, KD and Carlow, CK}, title = {Targeting the Wolbachia cell division protein FtsZ as a new approach for antifilarial therapy.}, journal = {PLoS neglected tropical diseases}, volume = {5}, number = {11}, pages = {e1411}, pmid = {22140592}, issn = {1935-2735}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/*antagonists & inhibitors/genetics/metabolism ; Berberine/*pharmacology ; Brugia malayi/*microbiology ; Cytoskeletal Proteins/*antagonists & inhibitors/genetics/metabolism ; DNA, Bacterial/chemistry/genetics ; Enzyme Inhibitors/pharmacology ; Filaricides/*pharmacology ; GTP Phosphohydrolases/*antagonists & inhibitors/genetics/metabolism ; Molecular Sequence Data ; Sequence Analysis, DNA ; Wolbachia/*drug effects ; }, abstract = {The use of antibiotics targeting the obligate bacterial endosymbiont Wolbachia of filarial parasites has been validated as an approach for controlling filarial infection in animals and humans. Availability of genomic sequences for the Wolbachia (wBm) present in the human filarial parasite Brugia malayi has enabled genome-wide searching for new potential drug targets. In the present study, we investigated the cell division machinery of wBm and determined that it possesses the essential cell division gene ftsZ which was expressed in all developmental stages of B. malayi examined. FtsZ is a GTPase thereby making the protein an attractive Wolbachia drug target. We described the molecular characterization and catalytic properties of Wolbachia FtsZ. We also demonstrated that the GTPase activity was inhibited by the natural product, berberine, and small molecule inhibitors identified from a high-throughput screen. Furthermore, berberine was also effective in reducing motility and reproduction in B. malayi parasites in vitro. Our results should facilitate the discovery of selective inhibitors of FtsZ as a novel anti-symbiotic approach for controlling filarial infection. NOTE: The nucleotide sequences reported in this paper are available in GenBank™ Data Bank under the accession number wAlB-FtsZ (JN616286).}, } @article {pmid22130021, year = {2011}, author = {Vavre, F and Mavingui, P}, title = {[Endosymbionts of arthropods and nematodes: allies to fight infectious diseases?].}, journal = {Medecine sciences : M/S}, volume = {27}, number = {11}, pages = {953-958}, doi = {10.1051/medsci/20112711010}, pmid = {22130021}, issn = {0767-0974}, mesh = {Animals ; Arthropods/genetics/*microbiology ; Bacteria/genetics/pathogenicity ; Bacterial Physiological Phenomena ; Communicable Diseases/*therapy ; Host-Parasite Interactions/genetics ; Humans ; Models, Biological ; Molecular Targeted Therapy/methods/trends ; Nematoda/genetics/*microbiology ; Symbiosis/genetics/*physiology ; }, abstract = {Arthropods and nematodes are important protagonists in human health because either they act as vectors of pathogens (bacteria, protozoa, viruses or fungus), or are themselves parasites. Fighting infectious diseases is based essentially on vaccination (prevention) or chemotherapeutic (curative) approaches in human, but one can envisage as an alternative to reduce the number of vectors or limit their ability to spread pathogens. Such strategies controlling dissemination will undoubtedly benefit from the knowledge accumulated by recent works on powerful mechanisms developed by symbiotic insect bacteria such as Wolbachia to popagate in arthropods and nematods. This review summarizes these recent data, and indicate how these mechanisms can be manipulated to reduce the dissemination of insect vectors propagating human diseases.}, } @article {pmid22126456, year = {2012}, author = {Taylor, M and Mediannikov, O and Raoult, D and Greub, G}, title = {Endosymbiotic bacteria associated with nematodes, ticks and amoebae.}, journal = {FEMS immunology and medical microbiology}, volume = {64}, number = {1}, pages = {21-31}, doi = {10.1111/j.1574-695X.2011.00916.x}, pmid = {22126456}, issn = {1574-695X}, mesh = {Amoeba/*microbiology/physiology ; Animals ; Bacterial Physiological Phenomena ; Nematoda/*microbiology/physiology ; *Symbiosis ; Ticks/*microbiology/physiology ; Wolbachia/*isolation & purification/physiology ; }, abstract = {Endosymbiosis is a mutualistic, parasitic or commensal symbiosis in which one symbiont is living within the body of another organism. Such symbiotic relationship with free-living amoebae and arthropods has been reported with a large biodiversity of microorganisms, encompassing various bacterial clades and to a lesser extent some fungi and viruses. By contrast, current knowledge on symbionts of nematodes is still mainly restricted to Wolbachia and its interaction with filarial worms that lead to increased pathogenicity of the infected nematode. In this review article, we aim to highlight the main characteristics of symbionts in term of their ecology, host cell interactions, parasitism and co-evolution, in order to stimulate future research in a field that remains largely unexplored despite the availability of modern tools.}, } @article {pmid22123956, year = {2012}, author = {Pan, X and Zhou, G and Wu, J and Bian, G and Lu, P and Raikhel, AS and Xi, Z}, title = {Wolbachia induces reactive oxygen species (ROS)-dependent activation of the Toll pathway to control dengue virus in the mosquito Aedes aegypti.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {109}, number = {1}, pages = {E23-31}, pmid = {22123956}, issn = {1091-6490}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; R37 AI024716/AI/NIAID NIH HHS/United States ; R37AI24716/AI/NIAID NIH HHS/United States ; R01AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/*microbiology/*virology ; Animals ; Animals, Genetically Modified ; Antioxidants/metabolism ; Cecropins/metabolism ; Defensins/metabolism ; Dengue/immunology/virology ; Dengue Virus/*physiology ; Fat Body/metabolism ; Gene Expression Regulation ; Gene Knockdown Techniques ; Gram-Negative Bacterial Infections/immunology/microbiology ; Humans ; Insect Proteins/genetics/metabolism ; Models, Immunological ; Organ Specificity ; Oxidative Stress ; Reactive Oxygen Species/*metabolism ; *Signal Transduction/genetics ; Toll-Like Receptors/*genetics/metabolism ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally transmitted symbiotic bacteria that can spread within insect populations because of their unique ability to manipulate host reproduction. When introduced to nonnative mosquito hosts, Wolbachia induce resistance to a number of human pathogens, including dengue virus (DENV), Plasmodium, and filarial nematodes, but the molecular mechanism involved is unclear. In this study, we have deciphered how Wolbachia infection affects the Aedes aegypti host in inducing resistance to DENV. The microarray assay indicates that transcripts of genes with functions related to immunity and reduction-oxidation (redox) reactions are up-regulated in Ae. aegypti infected with Wolbachia. Infection with this bacterium leads to induction of oxidative stress and an increased level of reactive oxygen species in its mosquito host. Reactive oxygen species elevation is linked to the activation of the Toll pathway, which is essential in mediating the expression of antioxidants to counterbalance oxidative stress. This immune pathway also is responsible for activation of antimicrobial peptides-defensins and cecropins. We provide evidence that these antimicrobial peptides are involved in inhibition of DENV proliferation in Wolbachia-infected mosquitoes. Utilization of transgenic Ae. aegypti and the RNAi depletion approach has been instrumental in proving the role of defensins and cecropins in the resistance of Wolbachia-infected Ae. aegypti to DENV. These results indicate that a symbiotic bacterium can manipulate the host defense system to facilitate its own persistent infection, resulting in a compromise of the mosquito's ability to host human pathogens. Our discoveries will aid in the development of control strategies for mosquito-transmitted diseases.}, } @article {pmid22123944, year = {2012}, author = {Blagrove, MS and Arias-Goeta, C and Failloux, AB and Sinkins, SP}, title = {Wolbachia strain wMel induces cytoplasmic incompatibility and blocks dengue transmission in Aedes albopictus.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {109}, number = {1}, pages = {255-260}, pmid = {22123944}, issn = {1091-6490}, support = {079059//Wellcome Trust/United Kingdom ; 095121//Wellcome Trust/United Kingdom ; /BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Aedes/genetics/immunology/*microbiology/*virology ; Animals ; Crosses, Genetic ; Cytoplasm/*microbiology ; Dengue/*transmission/*virology ; Dengue Virus/*physiology ; Drosophila melanogaster/microbiology ; Gene Expression Regulation ; Saliva/virology ; Wolbachia/*physiology ; }, abstract = {Wolbachia inherited bacteria are able to invade insect populations using cytoplasmic incompatibility and provide new strategies for controlling mosquito-borne tropical diseases, such as dengue. The overreplicating wMelPop strain was recently shown to strongly inhibit the replication of dengue virus when introduced into Aedes aegypti mosquitoes, as well as to stimulate chronic immune up-regulation. Here we show that stable introduction of the wMel strain of Drosophila melanogaster into Aedes albopictus, a vector of dengue and other arboviruses, abolished the transmission capacity of dengue virus-challenged mosquitoes. Immune up-regulation was observed in the transinfected line, but at a much lower level than that previously found for transinfected Ae. aegypti. Transient infection experiments suggest that this difference is related to Ae. albopictus immunotolerance of Wolbachia, rather than to the Wolbachia strain used. This study provides an example of strong pathogen inhibition in a naturally Wolbachia-infected mosquito species, demonstrating that this inhibition is not limited to naturally naïve species, and suggests that the Wolbachia strain is more important than host background for viral inhibition. Complete bidirectional cytoplasmic incompatibility was observed with WT strains infected with the naturally occurring Ae. albopictus Wolbachia, and this provides a mechanism for introducing wMel into natural populations of this species.}, } @article {pmid22116282, year = {2011}, author = {Ankola, K and Brueckner, D and Puttaraju, HP}, title = {Wolbachia endosymbiont infection in two Indian butterflies and female-biased sex ratio in the Red Pierrot, Talicada nyseus.}, journal = {Journal of biosciences}, volume = {36}, number = {5}, pages = {845-850}, pmid = {22116282}, issn = {0973-7138}, mesh = {Animals ; Butterflies/*microbiology/physiology ; Female ; Fertility ; Male ; Molecular Typing ; Sex Factors ; *Sex Ratio ; *Symbiosis ; *Wolbachia ; }, abstract = {The maternally inherited obligate bacteria Wolbachia is known to infect various lepidopteran insects. However, so far only a few butterfly species harbouring this bacterium have been thoroughly studied. The current study aims to identify the infection status of these bacteria in some of the commonly found butterfly species in India. A total of nine butterfly species belonging to four different families were screened using PCR with Wolbachia-specific wsp and ftsZ primers. The presence of the Wolbachia super group 'B' in the butterflies Red Pierrot, Talicada nyseus (Guerin) (Lepidoptera: Lycaenidae) and Blue Mormon, Papilio polymnestor Cramer (Papilionidae), is documented for the first time in India. The study also gives an account on the lifetime fecundity and female-biased sex ratio in T. nyseus, suggesting a putative role for Wolbachia in the observed female-biased sex ratio distortion.}, } @article {pmid22110767, year = {2011}, author = {Xue, HJ and Li, WZ and Nie, RE and Yang, XK}, title = {Recent speciation in three closely related sympatric specialists: inferences using multi-locus sequence, post-mating isolation and endosymbiont data.}, journal = {PloS one}, volume = {6}, number = {11}, pages = {e27834}, pmid = {22110767}, issn = {1932-6203}, mesh = {Animals ; Base Sequence ; Coleoptera/*genetics/microbiology/physiology ; Genes, Insect/genetics ; Genetic Loci/*genetics ; *Genetic Speciation ; Male ; *Phylogeny ; Plants ; *Reproductive Isolation ; Symbiosis/*genetics/physiology ; Sympatry/*genetics/physiology ; Time Factors ; Wolbachia/pathogenicity ; }, abstract = {Shifting between unrelated host plants is relatively rare for phytophagous insects, and distinct host specificity may play crucial roles in reproductive isolation. However, the isolation status and the relationship between parental divergence and post-mating isolation among closely related sympatric specialists are still poorly understood. Here, multi-locus sequence were used to estimate the relationship among three host plant-specific closely related flea beetles, Altica cirsicola, A. fragariae and A. viridicyanea (abbreviated as AC, AF and AV respectively). The tree topologies were inconsistent using different gene or different combinations of gene fragments. The relationship of AF+(AC+AV) was supported, however, by both gene tree and species tree based on concatenated data. Post-mating reproductive data on the results of crossing these three species are best interpreted in the light of a well established phylogeny. Nuclear-induced but not Wolbachia-induced unidirectional cytoplasmic incompatibility, which was detected in AC-AF and AF-AV but not in AC-AV, may also suggest more close genetic affinity between AC and AV. Prevalence of Wolbachia in these three beetles, and the endosymbiont in most individuals of AV and AC sharing a same wsp haplotype may give another evidence of AF+(AC+AV). Our study also suggested that these three flea beetles diverged in a relative short time (0.94 My), which may be the result of shifting between unrelated host plants and distinct host specificity. Incomplete post-mating isolation while almost complete lineage sorting indicated that effective pre-mating isolation among these three species should have evolved.}, } @article {pmid22100545, year = {2012}, author = {Abdul-Ghani, R and Al-Mekhlafi, AM and Alabsi, MS}, title = {Microbial control of malaria: biological warfare against the parasite and its vector.}, journal = {Acta tropica}, volume = {121}, number = {2}, pages = {71-84}, doi = {10.1016/j.actatropica.2011.11.001}, pmid = {22100545}, issn = {1873-6254}, mesh = {Animals ; Bacillus/pathogenicity ; *Biological Control Agents ; Culicidae/*microbiology/*virology ; Fungi/pathogenicity ; Malaria/*epidemiology/*prevention & control ; Pest Control, Biological/*methods ; Viruses/pathogenicity ; }, abstract = {Microbial applications in malaria transmission control have drawn global attention. Mosquito midgut microbiota can modulate vector immunity and block Plasmodium development. Paratransgenic manipulation of bacterial symbionts and Wolbachia can affect reproductive characteristics of mosquitoes. Bacillus-based biolarvicides can control mosquito larvae in different breeding habitats, but their effectiveness differs according to the type of formulation applied, and the physical and ecological conditions of the environment. Entomopathogenic fungi show promise as effective and evolution-proof agents against adult mosquitoes. In addition, transgenic fungi can express anti-plasmodial effector molecules that can target the parasite inside its vector. Despite showing effectiveness in domestic environments as well as against insecticide-resistant mosquitoes, claims towards their deployability in the field and their possible use in integrated vector management programmes have yet to be investigated. Viral pathogens show efficacy in the interruption of sporogonic development of the parasite, and protozoal pathogens exert direct pathogenic potential on larvae and adults with substantial effects on mosquito longevity and fecundity. However, the technology required for their isolation and maintenance impedes their field application. Many agents show promising findings; however, the question remains about the epidemiologic reality of these approaches because even those that have been tried under field conditions still have certain limitations. This review addresses aspects of the microbial control of malaria between proof-of-concept and epidemiologic reality.}, } @article {pmid22098559, year = {2012}, author = {Scott, AL and Ghedin, E and Nutman, TB and McReynolds, LA and Poole, CB and Slatko, BE and Foster, JM}, title = {Filarial and Wolbachia genomics.}, journal = {Parasite immunology}, volume = {34}, number = {2-3}, pages = {121-129}, pmid = {22098559}, issn = {1365-3024}, support = {Z01 AI000512-21/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Filariasis/parasitology ; Filarioidea/*genetics/microbiology ; Genome, Bacterial/genetics/physiology ; Genome, Helminth/*genetics/physiology ; Genomics/*methods ; Humans ; Proteomics ; RNA, Small Untranslated/genetics ; Symbiosis ; Transcriptome ; Wolbachia/*genetics ; }, abstract = {Filarial nematode parasites, the causative agents for a spectrum of acute and chronic diseases including lymphatic filariasis and river blindness, threaten the well-being and livelihood of hundreds of millions of people in the developing regions of the world. The 2007 publication on a draft assembly of the 95-Mb genome of the human filarial parasite Brugia malayi- representing the first helminth parasite genome to be sequenced - has been followed in rapid succession by projects that have resulted in the genome sequencing of six additional filarial species, seven nonfilarial nematode parasites of animals and nearly 30 plant parasitic and free-living species. Parallel to the genomic sequencing, transcriptomic and proteomic projects have facilitated genome annotation, expanded our understanding of stage-associated gene expression and provided a first look at the role of epigenetic regulation of filarial genomes through microRNAs. The expansion in filarial genomics will also provide a significant enrichment in our knowledge of the diversity and variability in the genomes of the endosymbiotic bacterium Wolbachia leading to a better understanding of the genetic principles that govern filarial-Wolbachia mutualism. The goal here is to provide an overview of the trends and advances in filarial and Wolbachia genomics.}, } @article {pmid22085419, year = {2011}, author = {Biliske, JA and Batista, PD and Grant, CL and Harris, HL}, title = {The bacteriophage WORiC is the active phage element in wRi of Drosophila simulans and represents a conserved class of WO phages.}, journal = {BMC microbiology}, volume = {11}, number = {}, pages = {251}, pmid = {22085419}, issn = {1471-2180}, mesh = {Animals ; Bacteriophages/*genetics/physiology ; DNA, Viral/genetics ; Drosophila/*microbiology ; Female ; Genome, Viral ; Larva/microbiology ; Male ; Phylogeny ; Prophages/*genetics/physiology ; Sequence Analysis, DNA ; Virus Replication ; Wolbachia/*virology ; }, abstract = {BACKGROUND: The alphaproteobacterium Wolbachia pipientis, the most common endosymbiont in eukaryotes, is found predominantly in insects including many Drosophila species. Although Wolbachia is primarily vertically transmitted, analysis of its genome provides evidence for frequent horizontal transfer, extensive recombination and numerous mobile genetic elements. The genome sequence of Wolbachia in Drosophila simulans Riverside (wRi) is available along with the integrated bacteriophages, enabling a detailed examination of phage genes and the role of these genes in the biology of Wolbachia and its host organisms. Wolbachia is widely known for its ability to modify the reproductive patterns of insects. One particular modification, cytoplasmic incompatibility, has previously been shown to be dependent on Wolbachia density and inversely related to the titer of lytic phage. The wRi genome has four phage regions, two WORiBs, one WORiA and one WORiC.

RESULTS: In this study specific primers were designed to distinguish between these four prophage types in wRi, and quantitative PCR was used to measure the titer of bacteriophages in testes, ovaries, embryos and adult flies. In all tissues tested, WORiA and WORiB were not found to be present in excess of their integrated prophages; WORiC, however, was found to be present extrachromosomally. WORiC is undergoing extrachromosomal replication in wRi. The density of phage particles was found to be consistent in individual larvae in a laboratory population. The WORiC genome is organized in conserved blocks of genes and aligns most closely with other known lytic WO phages, WOVitA and WOCauB.

CONCLUSIONS: The results presented here suggest that WORiC is the lytic form of WO in D. simulans, is undergoing extrachromosomal replication in wRi, and belongs to a conserved family of phages in Wolbachia.}, } @article {pmid22079163, year = {2012}, author = {Sabesan, S and Jambulingam, P}, title = {What ails Wolbachia transinfection to control disease vectors?.}, journal = {Trends in parasitology}, volume = {28}, number = {1}, pages = {1-2}, doi = {10.1016/j.pt.2011.10.004}, pmid = {22079163}, issn = {1471-5007}, mesh = {Animals ; Culicidae/microbiology/virology ; Dengue/*transmission ; Dengue Virus/*growth & development ; Insect Vectors/microbiology/virology ; Mosquito Control/methods ; Symbiosis ; *Transfection ; Transgenes ; Wolbachia/genetics/*physiology ; }, } @article {pmid22076370, year = {2011}, author = {James, S and Simmons, CP and James, AA}, title = {Ecology. Mosquito trials.}, journal = {Science (New York, N.Y.)}, volume = {334}, number = {6057}, pages = {771-772}, doi = {10.1126/science.1213798}, pmid = {22076370}, issn = {1095-9203}, mesh = {*Aedes/genetics/microbiology/physiology/virology ; Animals ; Animals, Genetically Modified ; Cluster Analysis ; Confounding Factors, Epidemiologic ; Dengue/epidemiology/*prevention & control/transmission ; Dengue Virus/physiology ; Genetic Engineering ; Humans ; *Insect Vectors/genetics/microbiology/physiology/virology ; *Pest Control, Biological ; Randomized Controlled Trials as Topic ; Wolbachia/physiology ; }, } @article {pmid22072969, year = {2011}, author = {Landmann, F and Voronin, D and Sullivan, W and Taylor, MJ}, title = {Anti-filarial activity of antibiotic therapy is due to extensive apoptosis after Wolbachia depletion from filarial nematodes.}, journal = {PLoS pathogens}, volume = {7}, number = {11}, pages = {e1002351}, pmid = {22072969}, issn = {1553-7374}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; *Apoptosis ; Brugia malayi/cytology/drug effects/metabolism/*microbiology ; Caspase 3/biosynthesis ; Doxycycline/pharmacology ; Female ; Filariasis/drug therapy/*parasitology ; Gerbillinae/parasitology ; Helminth Proteins/biosynthesis ; Humans ; In Situ Nick-End Labeling ; Larva/microbiology ; Male ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/*drug effects/*physiology ; }, abstract = {Filarial nematodes maintain a mutualistic relationship with the endosymbiont Wolbachia. Depletion of Wolbachia produces profound defects in nematode development, fertility and viability and thus has great promise as a novel approach for treating filarial diseases. However, little is known concerning the basis for this mutualistic relationship. Here we demonstrate using whole mount confocal microscopy that an immediate response to Wolbachia depletion is extensive apoptosis in the adult germline, and in the somatic cells of the embryos, microfilariae and fourth-stage larvae (L4). Surprisingly, apoptosis occurs in the majority of embryonic cells that had not been infected prior to antibiotic treatment. In addition, no apoptosis occurs in the hypodermal chords, which are populated with large numbers of Wolbachia, although disruption of the hypodermal cytoskeleton occurs following their depletion. Thus, the induction of apoptosis upon Wolbachia depletion is non-cell autonomous and suggests the involvement of factors originating from Wolbachia in the hypodermal chords. The pattern of apoptosis correlates closely with the nematode tissues and processes initially perturbed following depletion of Wolbachia, embryogenesis and long-term sterilization, which are sustained for several months until the premature death of the adult worms. Our observations provide a cellular mechanism to account for the sustained reductions in microfilarial loads and interruption of transmission that occurs prior to macrofilaricidal activity following antibiotic therapy of filarial nematodes.}, } @article {pmid22069350, year = {2011}, author = {Babalola, OE}, title = {Ocular onchocerciasis: current management and future prospects.}, journal = {Clinical ophthalmology (Auckland, N.Z.)}, volume = {5}, number = {}, pages = {1479-1491}, pmid = {22069350}, issn = {1177-5483}, abstract = {This paper reviews the current management of onchocerciasis and its future prospects. Onchocerciasis is a disease affecting millions of people in Africa, South and Central America, and Yemen. It is spread by the blackfly as a vector and caused by the filarial nematode, Onchocerca volvulus. A serious attempt was made by the Onchocerciasis Control Program between 1975 and 2002 to eliminate the vector in eleven of the endemic countries in West Africa, and with remarkable success. Formerly, the treatment was with diethyl carbamazine for the microfilaria and suramin for the adult worm. These drugs are now known to be toxic and unsuitable for mass distribution. In particular, they precipitate optic nerve disease. With the discovery of ivermectin, a much safer microfilaricide, and the decision of Merck to distribute the drug free of charge for as long as needed, the strategy of control switched to mass drug administration through community-directed treatment with ivermectin. So far, millions have received this annual or biannual treatment through the African Program for Onchocerciasis Control and the Onchocerciasis Elimination Program for the Americas. However, the problem with ivermectin is that it is a monotherapy microfilaricide which has limited effect on the adult worm, and thus will need to be continued for the life span of the adult worm, which may last up to 15 years. There are also early reports of resistance. Serious encephalopathy and death may occur when ivermectin is used in subjects heavily infested with loiasis. It seems unlikely that a break in transmission will occur with community-directed treatment with ivermectin in Africa because of population migrations and the highly efficient vector, but in the Americas some countries such as Columbia and the Oaxaca focus in Mexico have reported eradication. Vector control is only now applicable in selected situations, and particularly to control the nuisance value of the blackfly. Trials are ongoing for alternatives to ivermectin. Candidate drugs include moxidectin, a macrofilaricide, doxycycline which targets the Wolbachia endosymbiont, and flubendazole, which shows promise with the newer oral cyclodextrin formulation.}, } @article {pmid22069152, year = {2011}, author = {Berlanga, M and Paster, BJ and Grandcolas, P and Guerrero, R}, title = {Comparison of the gut microbiota from soldier and worker castes of the termite Reticulitermes grassei.}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {14}, number = {2}, pages = {83-93}, doi = {10.2436/20.1501.01.138}, pmid = {22069152}, issn = {1618-1905}, support = {DE-10374/DE/NIDCR NIH HHS/United States ; DE-11443/DE/NIDCR NIH HHS/United States ; }, mesh = {Animals ; Bacteria/*classification/*genetics ; *Biodiversity ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Gastrointestinal Tract/microbiology ; Isoptera/*microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {The bacterial microbiota from the whole gut of soldier and worker castes of the termite Reticulitermes grassei was isolated and studied. In addition, the 16S rDNA bacterial genes from gut DNA were PCR-amplified using Bacteria-selective primers, and the 16S rDNA amplicons subsequently cloned into Escherichia coli. Sequences of the cloned inserts were then used to determine closest relatives by comparison with published sequences and with sequences from our previous work. The clones were found to be affiliated with the phyla Spirochaetes, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Synergistetes, Verrucomicrobia, and candidate phyla Termite Group 1 (TG1) and Termite Group 2 (TG2). No significant differences were observed with respect to the relative bacterial abundances between soldier and worker phylotypes. The phylotypes obtained in this study were compared with reported sequences from other termites, especially those of phylotypes related to Spirochaetes, Wolbachia (an Alphaproteobacteria), Actinobacteria, and TG1. Many of the clone phylotypes detected in soldiers grouped with those of workers. Moreover, clones CRgS91 (soldiers) and CRgW68 (workers), both affiliated with 'Endomicrobia', were the same phylotype. Soldiers and workers also seemed to have similar relative protist abundances. Heterotrophic, poly-β-hydroxyalkanoate-accumulating bacteria were isolated from the gut of soldiers and shown to be affiliated with Actinobacteria and Gammaproteobacteria. We noted that Wolbachia was detected in soldiers but not in workers. Overall, the maintenance by soldiers and workers of comparable axial and radial redox gradients in the gut is consistent with the similarities in the prokaryotes and protists comprising their microbiota.}, } @article {pmid22068943, year = {2011}, author = {Marcon, HS and Domingues, DS and Coscrato, VE and Selivon, D and Perondini, AL and Marion, CL}, title = {New mariner elements in Anastrepha species (Diptera: Tephritidae).}, journal = {Neotropical entomology}, volume = {40}, number = {5}, pages = {568-574}, pmid = {22068943}, issn = {1678-8052}, mesh = {Animals ; DNA-Binding Proteins/*genetics ; Phylogeny ; Tephritidae/*classification/*genetics ; Transposases/*genetics ; }, abstract = {Mariner-like elements (MLE) are members from class II of transposable elements also known as DNA transposons. These elements have a wide distribution among different groups of organisms, including insects, which can be explained by horizontal and vertical gene-transfer. MLE families have been described in tephritid flies and other genera. During screening for Wolbachia bacteria in fruit flies of the genus Anastrepha, we discovered two sequences related to mariner-like elements. Based on these sequences, we designed primers that allowed us to isolate and characterize two new mariner-like elements (Anmar1 and Anmar2) in Anastrepha flies. These elements, which belong to the mellifera and rosa subfamilies have a low nucleotide diversity, and are probably inactive and acquired by vertical transfer. This is the first report of mariner-like transposons in flies found in South America.}, } @article {pmid22059383, year = {2011}, author = {Marsden, CD and Lee, Y and Nieman, CC and Sanford, MR and Dinis, J and Martins, C and Rodrigues, A and Cornel, AJ and Lanzaro, GC}, title = {Asymmetric introgression between the M and S forms of the malaria vector, Anopheles gambiae, maintains divergence despite extensive hybridization.}, journal = {Molecular ecology}, volume = {20}, number = {23}, pages = {4983-4994}, pmid = {22059383}, issn = {1365-294X}, support = {T32 AI074550-02/AI/NIAID NIH HHS/United States ; T32 AI074550/AI/NIAID NIH HHS/United States ; T32AI074550/AI/NIAID NIH HHS/United States ; R21AI062929/AI/NIAID NIH HHS/United States ; R21 AI062929/AI/NIAID NIH HHS/United States ; R21 AI062929-02/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*genetics/microbiology ; *Gene Flow ; Genetic Variation ; *Genetics, Population ; Genotyping Techniques ; Guinea-Bissau ; *Hybridization, Genetic ; Polymorphism, Single Nucleotide ; *Reproductive Isolation ; Sequence Analysis, DNA ; Wolbachia ; }, abstract = {The suggestion that genetic divergence can arise and/or be maintained in the face of gene flow has been contentious since first proposed. This controversy and a rarity of good examples have limited our understanding of this process. Partially reproductively isolated taxa have been highlighted as offering unique opportunities for identifying the mechanisms underlying divergence with gene flow. The African malaria vector, Anopheles gambiae s.s., is widely regarded as consisting of two sympatric forms, thought by many to represent incipient species, the M and S molecular forms. However, there has been much debate about the extent of reproductive isolation between M and S, with one view positing that divergence may have arisen and is being maintained in the presence of gene flow, and the other proposing a more advanced speciation process with little realized gene flow because of low hybrid fitness. These hypotheses have been difficult to address because hybrids are typically rare (<1%). Here, we assess samples from an area of high hybridization and demonstrate that hybrids are fit and responsible for extensive introgression. Nonetheless, we show that strong divergent selection at a subset of loci combined with highly asymmetric introgression has enabled M and S to remain genetically differentiated despite extensive gene flow. We propose that the extent of reproductive isolation between M and S varies across West Africa resulting in a 'geographic mosaic of reproductive isolation'; a finding which adds further complexity to our understanding of divergence in this taxon and which has considerable implications for transgenic control strategies.}, } @article {pmid22032328, year = {2012}, author = {McNulty, SN and Fischer, K and Mehus, JO and Vaughan, JA and Tkach, VV and Weil, GJ and Fischer, PU}, title = {Absence of Wolbachia endobacteria in Chandlerella quiscali, an avian filarial parasite.}, journal = {The Journal of parasitology}, volume = {98}, number = {2}, pages = {382-387}, pmid = {22032328}, issn = {1937-2345}, support = {R03 AI092306/AI/NIAID NIH HHS/United States ; T32 AI007172/AI/NIAID NIH HHS/United States ; T32-AI007172/AI/NIAID NIH HHS/United States ; R03-AI092306/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Antigens, Surface/immunology ; Bacterial Proteins/immunology ; Bird Diseases/*parasitology ; DNA, Bacterial/chemistry ; DNA, Helminth/chemistry ; DNA, Ribosomal/chemistry ; Female ; Filariasis/parasitology/*veterinary ; Filarioidea/classification/genetics/*microbiology ; Immunohistochemistry/veterinary ; In Situ Hybridization/veterinary ; Male ; Molecular Sequence Data ; Passeriformes/*parasitology ; Polymerase Chain Reaction/veterinary ; RNA, Bacterial/genetics ; RNA, Helminth/chemistry ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 5S/chemistry ; Wolbachia/genetics/immunology/*physiology ; }, abstract = {Chandlerella quiscali is a filarial nematode parasite of the common grackle (Quiscalus quiscula), a widespread bird species found throughout most of North America. Worms collected from wild-caught birds were morphologically identified as C. quiscali and tested for the presence of Wolbachia, an alphaproteobacterial endosymbiont required for reproduction and maturation by many filarial species. Although various methods, including polymerase chain reaction, in situ hybridization, and immunohistology, were used, we were unable to detect evidence of colonization with Wolbachia. Due to the widespread distribution of the grackle host, localization within the host, and high prevalence, C. quiscali may be among the most easily obtainable of Wolbachia-free filarial species. Further studies of C. quiscali and other Wolbachia-free filarial species may help to clarify the reason(s) that some filarial species require Wolbachia but others do not.}, } @article {pmid22022599, year = {2011}, author = {Verspoor, RL and Haddrill, PR}, title = {Genetic diversity, population structure and Wolbachia infection status in a worldwide sample of Drosophila melanogaster and D. simulans populations.}, journal = {PloS one}, volume = {6}, number = {10}, pages = {e26318}, pmid = {22022599}, issn = {1932-6203}, mesh = {Animals ; Drosophila/*genetics/*microbiology ; Drosophila melanogaster/*genetics/*microbiology ; Female ; *Genetic Variation ; Genetics, Population ; Gram-Negative Bacterial Infections/microbiology ; Microsatellite Repeats/genetics ; Symbiosis/genetics ; Wolbachia/*physiology ; }, abstract = {Drosophila melanogaster and its close relatives have been extremely important model species in the development of population genetic models that serve to explain patterns of diversity in natural populations, a major goal of evolutionary biology. A detailed picture of the evolutionary history of these species is beginning to emerge, as the relative importance of forces including demographic changes and natural selection is established. A continuing aim is to characterise levels of genetic diversity in a large number of populations of these species, covering a wide geographic area. We have used collections from five previously un-sampled wild populations of D. melanogaster and two of D. simulans, across three continents. We estimated levels of genetic diversity within, and divergence between, these populations, and looked for evidence of genetic structure both between ancestral and derived populations, and amongst derived populations. We also investigated the prevalence of infection with the bacterial endosymbiont Wolbachia. We found that D. melanogaster populations from Sub-Saharan Africa are the most diverse, and that divergence is highest between these and non-Sub-Saharan populations. There is strong evidence for structuring of populations between Sub-Saharan Africa and the rest of the world, and some evidence for weak structure amongst derived populations. Populations from Sub-Saharan Africa also differ in the prevalence of Wolbachia infection, with very low levels of infection compared to populations from the rest of the world.}, } @article {pmid22022422, year = {2011}, author = {Carpi, G and Cagnacci, F and Wittekindt, NE and Zhao, F and Qi, J and Tomsho, LP and Drautz, DI and Rizzoli, A and Schuster, SC}, title = {Metagenomic profile of the bacterial communities associated with Ixodes ricinus ticks.}, journal = {PloS one}, volume = {6}, number = {10}, pages = {e25604}, pmid = {22022422}, issn = {1932-6203}, mesh = {Animals ; Bacteria/classification/*genetics ; Base Sequence ; DNA, Complementary/genetics ; Genome, Bacterial/genetics ; Host-Pathogen Interactions/genetics ; Ixodes/*microbiology ; Metagenomics/*methods ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Software ; }, abstract = {Assessment of the microbial diversity residing in arthropod vectors of medical importance is crucial for monitoring endemic infections, for surveillance of newly emerging zoonotic pathogens, and for unraveling the associated bacteria within its host. The tick Ixodes ricinus is recognized as the primary European vector of disease-causing bacteria in humans. Despite I. ricinus being of great public health relevance, its microbial communities remain largely unexplored to date. Here we evaluate the pathogen-load and the microbiome in single adult I. ricinus by using 454- and Illumina-based metagenomic approaches. Genomic DNA-derived sequences were taxonomically profiled using a computational approach based on the BWA algorithm, allowing for the identification of known tick-borne pathogens at the strain level and the putative tick core microbiome. Additionally, we assessed and compared the bacterial taxonomic profile in nymphal and adult I. ricinus pools collected from two distinct geographic regions in Northern Italy by means of V6-16S rRNA amplicon pyrosequencing and community based ecological analysis. A total of 108 genera belonging to representatives of all bacterial phyla were detected and a rapid qualitative assessment for pathogenic bacteria, such as Borrelia, Rickettsia and Candidatus Neoehrlichia, and for other bacteria with mutualistic relationship or undetermined function, such as Wolbachia and Rickettsiella, was possible. Interestingly, the ecological analysis revealed that the bacterial community structure differed between the examined geographic regions and tick life stages. This finding suggests that the environmental context (abiotic and biotic factors) and host-selection behaviors affect their microbiome.Our data provide the most complete picture to date of the bacterial communities present within I. ricinus under natural conditions by using high-throughput sequencing technologies. This study further demonstrates a novel detection strategy for the microbiomes of arthropod vectors in the context of epidemiological and ecological studies.}, } @article {pmid22021671, year = {2011}, author = {Fast, EM and Toomey, ME and Panaram, K and Desjardins, D and Kolaczyk, ED and Frydman, HM}, title = {Wolbachia enhance Drosophila stem cell proliferation and target the germline stem cell niche.}, journal = {Science (New York, N.Y.)}, volume = {334}, number = {6058}, pages = {990-992}, pmid = {22021671}, issn = {1095-9203}, support = {K22 AI074909/AI/NIAID NIH HHS/United States ; 1K22AI74909-01A1/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Apoptosis ; Cell Proliferation ; Drosophila/*cytology/*microbiology/physiology ; Female ; Germ Cells/*cytology/physiology ; Male ; Mitosis ; Oogenesis ; Stem Cell Niche/*physiology ; Stem Cells/*cytology/physiology ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are widespread maternally transmitted intracellular bacteria that infect most insect species and are able to alter the reproduction of innumerous hosts. The cellular bases of these alterations remain largely unknown. Here, we report that Drosophila mauritiana infected with a native Wolbachia wMau strain produces about four times more eggs than the noninfected counterpart. Wolbachia infection leads to an increase in the mitotic activity of germline stem cells (GSCs), as well as a decrease in programmed cell death in the germarium. Our results suggest that up-regulation of GSC division is mediated by a tropism of Wolbachia for the GSC niche, the cellular microenvironment that supports GSCs.}, } @article {pmid22012236, year = {2011}, author = {de Albuquerque, AL and Magalhães, T and Ayres, CF}, title = {High prevalence and lack of diversity of Wolbachia pipientis in Aedes albopictus populations from Northeast Brazil.}, journal = {Memorias do Instituto Oswaldo Cruz}, volume = {106}, number = {6}, pages = {773-776}, doi = {10.1590/s0074-02762011000600021}, pmid = {22012236}, issn = {1678-8060}, mesh = {Aedes/*microbiology ; Animals ; *Genetic Variation/genetics ; Insect Vectors/*microbiology ; Polymerase Chain Reaction ; Prevalence ; Wolbachia/*genetics/isolation & purification ; }, abstract = {The use of Wolbachia as a tool to control insect vectors has recently been suggested. In this context, studies on the prevalence and diversity of this bacterium in wild populations are relevant. Here, we evaluated the diversity of two Wolbachiagenes (ftsZ and wsp) and the prevalence of this endosymbiont in wild Aedes albopictus. Using semi-nested polymerase chain reaction, our results showed that 99.3% of the individuals were superinfected with Wolbachia. In regards to genetic diversity, the two genes showed no variation within or among mosquito populations. An analysis of other Wolbachia markers may help to clarify the relationship between insect and endosymbiont.}, } @article {pmid21980455, year = {2011}, author = {Wong, ZS and Hedges, LM and Brownlie, JC and Johnson, KN}, title = {Wolbachia-mediated antibacterial protection and immune gene regulation in Drosophila.}, journal = {PloS one}, volume = {6}, number = {9}, pages = {e25430}, pmid = {21980455}, issn = {1932-6203}, mesh = {Animals ; Drosophila melanogaster/genetics/*immunology/*microbiology ; Gene Expression Regulation/*immunology ; Host-Pathogen Interactions/*genetics/*immunology ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Species Specificity ; Wolbachia/drug effects/growth & development/*immunology/pathogenicity ; }, abstract = {The outcome of microbial infection of insects is dependent not only on interactions between the host and pathogen, but also on the interactions between microbes that co-infect the host. Recently the maternally inherited endosymbiotic bacteria Wolbachia has been shown to protect insects from a range of microbial and eukaryotic pathogens. Mosquitoes experimentally infected with Wolbachia have upregulated immune responses and are protected from a number of pathogens including viruses, bacteria, Plasmodium and filarial nematodes. It has been hypothesised that immune upregulation underpins Wolbachia-mediated protection. Drosophila is a strong model for understanding host-Wolbachia-pathogen interactions. Wolbachia-mediated antiviral protection in Drosophila has been demonstrated for a number of different Wolbachia strains. In this study we investigate whether Wolbachia-infected flies are also protected against pathogenic bacteria. Drosophila simulans lines infected with five different Wolbachia strains were challenged with the pathogenic bacteria Pseudomonas aeruginosa PA01, Serratia marcescens and Erwinia carotovora and mortality compared to paired lines without Wolbachia. No difference in mortality was observed in the flies with or without Wolbachia. Similarly no antibacterial protection was observed for D. melanogaster infected with Wolbachia. Interestingly, D. melanogaster Oregon RC flies which are naturally infected with Wolbachia showed no upregulation of the antibacterial immune genes TepIV, Defensin, Diptericin B, PGRP-SD, Cecropin A1 and Attacin D compared to paired flies without Wolbachia. Taken together these results indicate that Wolbachia-mediated antibacterial protection is not ubiquitous in insects and furthermore that the mechanisms of antibacterial and antiviral protection are independent. We suggest that the immune priming and antibacterial protection observed in Wolbachia-infected mosquitoes may be a consequence of the recent artificial introduction of the symbiont into insects that normally do not carry Wolbachia and that antibacterial protection is unlikely to be found in insects carrying long-term Wolbachia infections.}, } @article {pmid21975225, year = {2011}, author = {Crain, PR and Mains, JW and Suh, E and Huang, Y and Crowley, PH and Dobson, SL}, title = {Wolbachia infections that reduce immature insect survival: predicted impacts on population replacement.}, journal = {BMC evolutionary biology}, volume = {11}, number = {}, pages = {290}, pmid = {21975225}, issn = {1471-2148}, support = {AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Computer Simulation ; Female ; *Host-Pathogen Interactions ; Insecta/*microbiology ; Larva/microbiology ; Male ; *Models, Biological ; Population Dynamics ; *Symbiosis ; *Wolbachia ; }, abstract = {BACKGROUND: The evolutionary success of Wolbachia bacteria, infections of which are widespread in invertebrates, is largely attributed to an ability to manipulate host reproduction without imposing substantial fitness costs. Here, we describe a stage-structured model with deterministic immature lifestages and a stochastic adult female lifestage. Simulations were conducted to better understand Wolbachia invasions into uninfected host populations. The model includes conventional Wolbachia parameters (the level of cytoplasmic incompatibility, maternal inheritance, the relative fecundity of infected females, and the initial Wolbachia infection frequency) and a new parameter termed relative larval viability (RLV), which is the survival of infected larvae relative to uninfected larvae.

RESULTS: The results predict the RLV parameter to be the most important determinant for Wolbachia invasion and establishment. Specifically, the fitness of infected immature hosts must be close to equal to that of uninfected hosts before population replacement can occur. Furthermore, minute decreases in RLV inhibit the invasion of Wolbachia despite high levels of cytoplasmic incompatibility, maternal inheritance, and low adult fitness costs.

CONCLUSIONS: The model described here takes a novel approach to understanding the spread of Wolbachia through a population with explicit dynamics. By combining a stochastic female adult lifestage and deterministic immature/adult male lifestages, the model predicts that even those Wolbachia infections that cause minor decreases in immature survival are unlikely to invade and spread within the host population. The results are discussed in relation to recent theoretical and empirical studies of natural population replacement events and proposed applied research, which would use Wolbachia as a tool to manipulate insect populations.}, } @article {pmid21966105, year = {2010}, author = {Mahajan, RS and Veerpathran, A and Dakshinamoorthy, G and Sharma, RD and Goswami, K and Reddy, MV}, title = {Effect of Certain Antibiotics Against Filarial Parasite Brugia malayi In Vitro: Possible Role of Oxidative Stress.}, journal = {Indian journal of clinical biochemistry : IJCB}, volume = {25}, number = {4}, pages = {362-366}, pmid = {21966105}, issn = {0974-0422}, abstract = {WHO-Tropical Disease Research scheme highlighted the need for development of new anti-filarial drugs. Certain antibiotics have recently been found effective against Wolbachia, co-existing symbiotically with filarial parasites. Inflammatory response entails oxidative mechanism to educe direct anti-microbial effect. In the present study microfilariae were maintained in vitro in medium supplemented with varying concentrations of tetracycline, doxycycline (20-100 μg/ml) or ciprofloxacin (50-250 μg/ml) separately to find out any involvement of oxidative mechanism in the anti-filarial effect of these antibiotics. Loss of motility of the microfilariae was measured after 48 h and correlated with the levels of MDA, nitric oxide and protein-carbonylation. Significant loss of microfilarial motility was recorded with increasing concentration of tetracycline and doxycycline but with ciprofloxacin the effect was not marked. Agents with high antifilarial activity revealed significant association with oxidative parameters in a dose dependent manner. The result suggests that oxidative effect might be exploited to design novel antifilarial drug candidate.}, } @article {pmid21949820, year = {2011}, author = {Kent, BN and Funkhouser, LJ and Setia, S and Bordenstein, SR}, title = {Evolutionary genomics of a temperate bacteriophage in an obligate intracellular bacteria (Wolbachia).}, journal = {PloS one}, volume = {6}, number = {9}, pages = {e24984}, pmid = {21949820}, issn = {1932-6203}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Bacteriophages/*genetics ; *Evolution, Molecular ; Genes, Bacterial ; Genome, Bacterial ; *Genomics ; Lysogeny ; Phylogeny ; Recombination, Genetic ; Selection, Genetic ; Wolbachia/*genetics ; }, abstract = {Genome evolution of bacteria is usually influenced by ecology, such that bacteria with a free-living stage have large genomes and high rates of horizontal gene transfer, while obligate intracellular bacteria have small genomes with typically low amounts of gene exchange. However, recent studies indicate that obligate intracellular species that host-switch frequently harbor agents of horizontal transfer such as mobile elements. For example, the temperate double-stranded DNA bacteriophage WO in Wolbachia persistently transfers between bacterial coinfections in the same host. Here we show that despite the phage's rampant mobility between coinfections, the prophage's genome displays features of constraint related to its intracellular niche. First, there is always at least one intact prophage WO and usually several degenerate, independently-acquired WO prophages in each Wolbachia genome. Second, while the prophage genomes are modular in composition with genes of similar function grouping together, the modules are generally not interchangeable with other unrelated phages and thus do not evolve by the Modular Theory. Third, there is an unusual core genome that strictly consists of head and baseplate genes; other gene modules are frequently deleted. Fourth, the prophage recombinases are diverse and there is no conserved integration sequence. Finally, the molecular evolutionary forces acting on prophage WO are point mutation, intragenic recombination, deletion, and purifying selection. Taken together, these analyses indicate that while lateral transfer of phage WO is pervasive between Wolbachia with occasional new gene uptake, constraints of the intracellular niche obstruct extensive mixture between WO and the global phage population. Although the Modular Theory has long been considered the paradigm of temperate bacteriophage evolution in free-living bacteria, it appears irrelevant in phages of obligate intracellular bacteria.}, } @article {pmid21946455, year = {2012}, author = {Wu, K and Hoy, MA}, title = {Extended starvation reduced and eliminated Wolbachia, but not Cardinium, from Metaseiulus occidentalis females (Acari: Phytoseiidae): a need to reassess Wolbachia's status in this predatory mite?.}, journal = {Journal of invertebrate pathology}, volume = {109}, number = {1}, pages = {20-26}, doi = {10.1016/j.jip.2011.09.005}, pmid = {21946455}, issn = {1096-0805}, mesh = {Animals ; Bacterial Load ; Bacteroidetes/*physiology ; DNA, Bacterial/isolation & purification ; Female ; *Host-Pathogen Interactions ; Mites/*microbiology/physiology ; Ovum/microbiology ; Pest Control, Biological ; Polymerase Chain Reaction/methods ; Starvation/*physiopathology ; Symbiosis/physiology ; Wolbachia/*physiology ; }, abstract = {The presence of Wolbachia and Cardinium bacteria has been documented in many arthropod species, including the predatory mite Metaseiulus (=Typhlodromus or Galendomus) occidentalis (Nesbitt) (Acari: Phytoseiidae). We show that Tetranychus urticae, the prey of Metaseiulus occidentalis, contains Wolbachia and no detectable Cardinium using quantitative PCR (qPCR). Starvation for 72 h at 22°C eliminated most, if not all, Wolbachia in M. occidentalis adult females from 7 laboratory colonies. Refeeding of M. occidentalis with T. urticae after starvation for 72 h restored the amounts of Wolbachia in M. occidentalis to those of prestarvation levels, suggesting that Wolbachia detected in M. occidentalis starved for shorter periods of time in current, and some previous, studies likely came from T. urticae. Furthermore, eggs from all M. occidentalis colonies examined were free of Wolbachia if they were surface-decontaminated with 0.3% sodium hypochlorite before DNA extraction. Cardinium was present in 6 of 14 laboratory colonies of M. occidentalis. Starvation for 3, 24, 48, and 72 h had no effect on the amounts of Cardinium in adult females from the Cardinium-positive colonies. Eggs from these colonies were positive for Cardinium but contained less than 1% of the titers found in adult females. The data suggest that Cardinium, but not Wolbachia, is an endosymbiont in certain populations of M. occidentalis. In light of our current findings, we recommend specific practices for the identification of potential symbionts in predatory arthropod species using the PCR.}, } @article {pmid21945051, year = {2012}, author = {Müller, MJ and von Mühlen, C and Valiati, VH and da Silva Valente, VL}, title = {Wolbachia pipientis is associated with different mitochondrial haplotypes in natural populations of Drosophila willistoni.}, journal = {Journal of invertebrate pathology}, volume = {109}, number = {1}, pages = {152-155}, doi = {10.1016/j.jip.2011.08.011}, pmid = {21945051}, issn = {1096-0805}, mesh = {Animals ; Brazil ; DNA, Mitochondrial/genetics ; Drosophila/genetics/*microbiology ; Evolution, Molecular ; Gene Transfer, Horizontal ; Genetic Variation ; Gram-Negative Bacterial Infections/microbiology/*veterinary ; Haplotypes ; Host-Pathogen Interactions ; Mitochondria/*genetics/microbiology ; Pest Control, Biological ; Symbiosis ; Wolbachia/*pathogenicity/physiology ; }, abstract = {The prevalence of the endosymbiont Wolbachia pipientis and its effects on mitochondrial genetic diversity were analyzed in natural populations of Drosophila willistoni, a neotropical species recently infected. Total infection rate was 55% and no evidence was found that the Wolbachia infection decreased the diversity of mtDNA. Wolbachia was seen to be associated with different mitochondria, suggesting multiple horizontal transmission events and/or transmission paternal leakage of mitochondrial and/or Wolbachia. These hypotheses are evaluated in the context of the present study and other research.}, } @article {pmid21944995, year = {2011}, author = {Liu, C and Kelen, PV and Ghedin, E and Lustigman, S and Unnasch, TR}, title = {Analysis of transcriptional regulation of tetracycline responsive genes in Brugia malayi.}, journal = {Molecular and biochemical parasitology}, volume = {180}, number = {2}, pages = {106-111}, pmid = {21944995}, issn = {1872-9428}, support = {R01 AI072465/AI/NIAID NIH HHS/United States ; R01 AI072465-04/AI/NIAID NIH HHS/United States ; 1R01AI072465/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Base Sequence ; Brugia malayi/*genetics/metabolism ; *Gene Expression Regulation ; Helminth Proteins/*genetics/metabolism ; Molecular Sequence Data ; *Response Elements ; TATA Box ; Tetracycline/*metabolism ; *Transcription, Genetic ; }, abstract = {The Wolbachia endosymbiont of the human filarial parasites is necessary for parasite reproduction, making it an attractive chemotherapeutic target. Previous studies have demonstrated that mRNA levels of several nuclearly encoded genes are altered as a result of exposure to antibiotics that eliminate the endosymbiont, suggesting that they may be involved in maintaining the parasite-endosymbiont relationship. Here, we tested the hypothesis that the increase in mRNA levels of certain nuclearly encoded genes of Brugia malayi in response to tetracycline treatment involved specific regulatory elements present in the promoters of these genes. The promoters of three such genes (BmRPL13, BmRPS4 and BmHSP70) were tested for tetracycline responsiveness utilizing a homologous transient transcription system. Reporter gene expression driven by all three promoters was up-regulated in transfected embryos exposed to tetracycline. Substitution mutagenesis was employed to map the cis-acting elements responsible for this response in the BmHSP70 promoter. Tetracycline responsiveness was found to be distinct from the cis-acting elements involved in regulating the stress response from the BmHSP70 promoter; rather, tetracycline responsiveness was mediated by a TATAA-box like element. This study represents the first demonstration of small molecule-mediated gene regulation of a native B. malayi promoter.}, } @article {pmid21940637, year = {2011}, author = {Cerveau, N and Leclercq, S and Leroy, E and Bouchon, D and Cordaux, R}, title = {Short- and long-term evolutionary dynamics of bacterial insertion sequences: insights from Wolbachia endosymbionts.}, journal = {Genome biology and evolution}, volume = {3}, number = {}, pages = {1175-1186}, pmid = {21940637}, issn = {1759-6653}, mesh = {Base Sequence ; *DNA Transposable Elements ; *Evolution, Molecular ; *Genome, Bacterial ; Phylogeny ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Transposable elements (TE) are one of the major driving forces of genome evolution, raising the question of the long-term dynamics underlying their evolutionary success. Long-term TE evolution can readily be reconstructed in eukaryotes, thanks to many degraded copies constituting genomic fossil records of past TE proliferations. By contrast, bacterial genomes usually experience high sequence turnover and short TE retention times, thereby obscuring ancient TE evolutionary patterns. We found that Wolbachia bacterial genomes contain 52-171 insertion sequence (IS) TEs. IS account for 11% of Wolbachia wRi, which is one of the highest IS genomic coverage reported in prokaryotes to date. We show that many IS groups are currently expanding in various Wolbachia genomes and that IS horizontal transfers are frequent among strains, which can explain the apparent synchronicity of these IS proliferations. Remarkably, >70% of Wolbachia IS are nonfunctional. They constitute an unusual bacterial IS genomic fossil record providing direct empirical evidence for a long-term IS evolutionary dynamics following successive periods of intense transpositional activity. Our results show that comprehensive IS annotations have the potential to provide new insights into prokaryote TE evolution and, more generally, prokaryote genome evolution. Indeed, the identification of an important IS genomic fossil record in Wolbachia demonstrates that IS elements are not always of recent origin, contrary to the conventional view of TE evolution in prokaryote genomes. Our results also raise the question whether the abundance of IS fossils is specific to Wolbachia or it may be a general, albeit overlooked, feature of prokaryote genomes.}, } @article {pmid21936319, year = {2011}, author = {Brelsfoard, CL and Dobson, SL}, title = {Wolbachia effects on host fitness and the influence of male aging on cytoplasmic incompatibility in Aedes polynesiensis (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {48}, number = {5}, pages = {1008-1015}, doi = {10.1603/me10202}, pmid = {21936319}, issn = {0022-2585}, support = {R01-AI067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/growth & development/*microbiology/*physiology ; Aging ; Animals ; Cytoplasm/microbiology ; Female ; Fertility ; Genetic Fitness ; Host-Pathogen Interactions ; Larva/growth & development/microbiology/physiology ; Longevity ; Male ; *Pest Control, Biological ; Polymerase Chain Reaction ; Population Dynamics ; Sex Ratio ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacteria Wolbachia manipulate host reproduction by inducing a form of sterility known as cytoplasmic incompatibility (CI), promoting the invasion of infection into natural host populations. CI has received attention for use in applied strategies to control insect vectors of disease. Thus, to understand both naturally occurring Wolbachia invasions and evaluate potential applied strategies, it is important to understand Wolbachia interactions with its host, including impacts on fitness and the CI level. In this study, we examined for an effect of Wolbachia on survivorship, developmental time, sex ratio, longevity, fecundity, and egg hatch of Aedes polynesiensis Marks, which is the primary vector of Wuchereria bancrofti in the South Pacific. In this study, we have compared strains of A. polynesiensis that are naturally and artificially infected with Wolbachia and additional strains that are aposymbiotic (Wolbachia removed to generate an uninfected strain). Artificially infected strains were observed to have increased larval mortality and decreased adult longevity when compared with aposymbiotic strains. Naturally infected strains were observed to have decreased larval mortality, pupal mortality, increased adult longevity, and a larger adult size when compared with aposymbiotic strains. Artificially infected males that were 4 wk old were able to induce high rates of CI, similar to young males. We discuss the results in relation to the natural spread of Wolbachia and Wolbachia-based applied strategies to modify A. polynesiensis populations.}, } @article {pmid21936318, year = {2011}, author = {Endersby, NM and Hoffmann, AA and White, VL and Ritchie, SA and Johnson, PH and Weeks, AR}, title = {Changes in the genetic structure of Aedes aegypti (Diptera: Culicidae) populations in Queensland, Australia, across two seasons: implications for potential mosquito releases.}, journal = {Journal of medical entomology}, volume = {48}, number = {5}, pages = {999-1007}, pmid = {21936318}, issn = {0022-2585}, support = {R29 AI034355-03/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/classification/*genetics/microbiology/virology ; Animals ; Dengue/*prevention & control ; Dengue Virus/*physiology ; Genetic Variation ; Insect Vectors/classification/*genetics/microbiology/virology ; *Pest Control, Biological ; Population Dynamics ; Queensland ; Seasons ; Wolbachia/*physiology ; }, abstract = {Diseases transmitted by mosquitoes could be controlled if vector populations were replaced with strains that have reduced vector competency. Such a strategy is being developed for control of dengue virus which is transmitted by Aedes aegypti (L.) (Diptera: Culicidae). Mosquitoes artificially infected with the bacterium, Wolbachia pipientis Hertig, are being assessed as candidates for release at the adult stage with the aim of replacement of the wild population. Wolbachia can reduce the capacity of Ae. aegypti to transmit dengue virus and has potential to be driven through the natural population via a system of cytoplasmic incompatibility. Deployment of benign mosquito strains will be influenced by population size and structure of wild-type Ae. aegypti in proposed release areas, as well as rates of gene flow among populations in the wet and dry tropical seasons. Mosquitoes from northern Queensland were screened with genetic markers to find an optimal locality for release of a benign strain of Ae. aegypti. The inland towns of Chillagoe and Charters Towers and the coastal town of Ingham had mosquito populations that were partly genetically isolated from mosquitoes in other areas across both seasons. These locations may be suitable release sites if it is important for the released strain to be restricted during initial phases of implementation. Smaller genetic differences were also evident among other regions and were consistent over two seasons (wet and dry).}, } @article {pmid21934703, year = {2012}, author = {Reumer, BM and van Alphen, JJ and Kraaijeveld, K}, title = {Occasional males in parthenogenetic populations of Asobara japonica (Hymenoptera: Braconidae): low Wolbachia titer or incomplete coadaptation?.}, journal = {Heredity}, volume = {108}, number = {3}, pages = {341-346}, pmid = {21934703}, issn = {1365-2540}, mesh = {Animals ; DNA, Mitochondrial ; Female ; Haplotypes ; Host-Pathogen Interactions ; Hymenoptera/*genetics/*microbiology ; Male ; Molecular Sequence Data ; Parthenogenesis/*genetics ; Reproduction/genetics ; Sex Ratio ; Symbiosis ; Temperature ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria known to manipulate the reproduction of their hosts. Some populations of the parasitoid wasp Asobara japonica are infected with Wolbachia and reproduce parthenogenetically, while other populations are not infected and reproduce sexually. Wolbachia-infected A. japonica females regularly produce small numbers of male offspring. Because all females in the field are infected and infected females are not capable of sexual reproduction, male production seems to be maladaptive. We investigated why these females nevertheless produce males. We tested three hypotheses: high rearing temperatures could result in higher offspring sex ratios (more males), low Wolbachia titer of the mother could lead to higher offspring sex ratios and/or the Wolbachia infection is of relatively recent origin and not enough time has passed to allow complete coadaptation between Wolbachia and host. In all, 33% of the Wolbachia-infected females produced males and 56% of these males were also infected with Wolbachia. Neither offspring sex ratio nor male infection frequency was significantly affected by rearing temperature or Wolbachia concentration of the mother. The mitochondrial DNA sequence of one of the uninfected populations was identical to that of two of the infected populations. Therefore, the initial Wolbachia infection of A. japonica must have occurred recently. Mitochondrial sequence variation among the infected populations suggests that the spread of Wolbachia through the host populations involved horizontal transmission. We conclude that the occasional male production by Wolbachia-infected females is most likely a maladaptive side effect of incomplete coevolution between symbiont and host in this relatively young infection.}, } @article {pmid21923674, year = {2012}, author = {Henri, H and Mouton, L}, title = {High-resolution melting technology: a new tool for studying the Wolbachia endosymbiont diversity in the field.}, journal = {Molecular ecology resources}, volume = {12}, number = {1}, pages = {75-81}, doi = {10.1111/j.1755-0998.2011.03064.x}, pmid = {21923674}, issn = {1755-0998}, mesh = {Animals ; Arthropods/*microbiology/physiology ; *Biodiversity ; *Genetic Techniques ; Phylogeny ; *Symbiosis ; Transition Temperature ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Stable infections by maternally transmitted symbionts are frequently found in field populations, especially in arthropods. Many questions remain regarding their contribution to host biology and ecology, and especially on environmental adaptation of their host. Wolbachia is one of the most common endosymbiont of invertebrates. This cytoplasmically inherited endocellular bacterium induces number of reproductive alterations in its arthropod hosts and various fitness effects that allow it to spread in host populations. To better understand the influence of Wolbachia on host phenotypes and consequences of the manipulation of reproduction on the host genetic differentiation, it is crucial to be able to discriminate Wolbachia strains and determine their prevalence, which requires exhaustive screening. In the present report, we proposed the use of a new tool for the population studies, based on the high resolution melting (HRM) analysis, less expensive and faster than the 'classical' methods for large-scale studies. We investigated the effectiveness of HRM to explore and characterize the diversity of Wolbachia strains. Results obtained showed that HRM is a powerful tool to identify strains and detect polymorphism in singly infected hosts. When individuals harboured a mixture of Wolbachia strains (multiple infections), there is a risk of underestimation of the diversity if the proportions of the strains are highly different. However, the same limitations exist for the other techniques commonly used. Overall, this study demonstrated that HRM analysis is a rapid and reliable technique useful for studying, without a priori, Wolbachia strains diversity in field populations.}, } @article {pmid21922235, year = {2012}, author = {McHaffie, J}, title = {Dirofilaria immitis and Wolbachia pipientis: a thorough investigation of the symbiosis responsible for canine heartworm disease.}, journal = {Parasitology research}, volume = {110}, number = {2}, pages = {499-502}, pmid = {21922235}, issn = {1432-1955}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Dirofilaria immitis/*microbiology/*pathogenicity/physiology ; Dirofilariasis/drug therapy/*parasitology ; Dog Diseases/drug therapy/*parasitology ; Dogs ; Doxycycline/therapeutic use ; *Symbiosis ; Treatment Outcome ; United States ; Wolbachia/*physiology ; }, abstract = {Canine heartworm disease wreaks havoc inside canines all throughout the modern world, including the USA. Any region where mosquitoes thrive will provide efficient dog-to-dog transportation for the microfilaria of the infectious nematode Dirofilaria immitis. Veterinary scientists have recently discovered both phylogenetic and biochemical evidence for the obligate symbiosis of D. immitis and the bacteria Wolbachia pipientis. As a result, veterinarians have initiated testing of antibiotic therapies either instead of, or together with, the currently utilized antiparasitic treatments for canine heartworm. The toxicity of melarsomine adulticidal therapies has prompted an abundance of new research involving doxycycline and other antibiotics, which will be addressed in this review. As our knowledge of the Wolbachia endosymbiont expands, so will our abilities to minimize toxicity and maximize efficiency of heartworm treatments.}, } @article {pmid21919732, year = {2012}, author = {Morchón, R and Carretón, E and Grandi, G and González-Miguel, J and Montoya-Alonso, JA and Simón, F and Genchi, C and Kramer, LH}, title = {Anti-Wolbachia Surface Protein antibodies are present in the urine of dogs naturally infected with Dirofilaria immitis with circulating microfilariae but not in dogs with occult infections.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {12}, number = {1}, pages = {17-20}, pmid = {21919732}, issn = {1557-7759}, mesh = {Animals ; Antibodies, Bacterial/*urine ; Bacterial Outer Membrane Proteins/*immunology ; *Dirofilaria immitis/immunology/microbiology ; Dirofilariasis/blood/*immunology/urine ; Dog Diseases/blood/*immunology/urine ; Dogs ; Microfilariae/immunology/microbiology ; Wolbachia/*immunology ; }, abstract = {Heartworm infection (Dirofilaria immitis) can cause kidney damage due to the presence of circulating microfilariae (mf) that contribute to the production and deposit of immune complexes. It has been shown that mf are a major source of Wolbachia antigen during active infection. Here the authors compared urine samples from 19 naturally infected dogs with (mf+) and 12 without (mf-) microfilariae for the presence of proteinuria and anti-Wolbachia Surface Protein (-WSP) IgG in ELISA. Kidneys from 6 mf+ and 3 mf- dogs were also examined by anti-WSP immuno-histochemistry. All infected dogs showed proteinuria, but mf+ dogs had significantly higher values compared to mf-dogs. Mf+ dogs had optical density values for anti-WSP IgG consistently higher than established cut-off values and were significantly higher than values for mf- dogs. Kidneys from mf+ dogs showed Wolbachia+ mf in glomerular capillaries. Results strongly suggest that Wolbachia associated with circulating mf may contribute to immune-mediated kidney disease in dogs with heartworm infection.}, } @article {pmid21907309, year = {2011}, author = {Almeida, Fd and Moura, AS and Cardoso, AF and Winter, CE and Bijovsky, AT and Suesdek, L}, title = {Effects of Wolbachia on fitness of Culex quinquefasciatus (Diptera; Culicidae).}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {11}, number = {8}, pages = {2138-2143}, doi = {10.1016/j.meegid.2011.08.022}, pmid = {21907309}, issn = {1567-7257}, mesh = {Animals ; Crosses, Genetic ; Culex/anatomy & histology/genetics/*microbiology/physiology ; Female ; *Genetic Fitness ; Longevity ; Male ; Oviposition ; Reproduction/physiology ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia are α-proteobacteria that were first reported in Culex pipiens mosquitoes early in the twentieth century. Since then, the effect of Wolbachia on their host's reproduction has drawn attention and has been increasingly investigated. Given the extreme complexity of this interaction, new study cases are welcomed to enhance its understanding. The present work addressed the influence of Wolbachia on Cx. quinquefasciatus, the cosmopolitan member of the Cx. pipiens complex. Samples of a Cx. quinquefasciatus colony (wPip(+)) originated from individuals naturally infected by Wolbachiapipientis B strain, were cured with tetracycline, yielding a Wolbachia-free colony (wPip(-)). Both the presence of bacteria and the efficiency of bacterial elimination were checked by PCR of the wsp gene. Total reproductive unidirectional incompatibility occurred when wPip(-) females were crossed with wPip(+) males, whereas the other three types of reciprocal crosses were viable. Reproductive aspects were also comparatively evaluated between colonies. Concerning oviposition time during the first gonotrophic cycle, wPip(+) females developed and laid eggs earlier than did wPip(-) females. Reproductive fitness was higher among wPip(-) than wPip(+) females regarding the following parameters: fertility: egg rafts/fed females; fecundity: eggs/raft, and viability: larvae/eggs. Conversely, longevity of wPip(-) females was lower. Summarising, although the infected mosquitoes have the advantage of a higher longevity, they have lower reproductive fitness. Our results are partly distinct from all other reports on Aedes and Culex mosquitoes previously published.}, } @article {pmid21881776, year = {2011}, author = {Walker, T and Moreira, LA}, title = {Can Wolbachia be used to control malaria?.}, journal = {Memorias do Instituto Oswaldo Cruz}, volume = {106 Suppl 1}, number = {}, pages = {212-217}, doi = {10.1590/s0074-02762011000900026}, pmid = {21881776}, issn = {1678-8060}, mesh = {Aedes/*microbiology ; Animals ; Humans ; Insect Vectors/*microbiology ; Malaria/*prevention & control ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, abstract = {Malaria is a mosquito-borne infectious disease caused by Plasmodium parasites transmitted by the infectious bite of Anopheles mosquitoes. Vector control of malaria has predominantly focused on targeting the adult mosquito through insecticides and bed nets. However, current vector control methods are often not sustainable for long periods so alternative methods are needed. A novel biocontrol approach for mosquito-borne diseases has recently been proposed, it uses maternally inherited endosymbiotic Wolbachia bacteria transinfected into mosquitoes in order to interfere with pathogen transmission. Transinfected Wolbachia strains in Aedes aegypti mosquitoes, the primary vector of dengue fever, directly inhibit pathogen replication, including Plasmodium gallinaceum, and also affect mosquito reproduction to allow Wolbachia to spread through mosquito populations. In addition, transient Wolbachia infections in Anopheles gambiae significantly reduce Plasmodium levels. Here we review the prospects of using a Wolbachia-based approach to reduce human malaria transmission through transinfection of Anopheles mosquitoes.}, } @article {pmid21884292, year = {2011}, author = {Kremer, N and Huigens, ME}, title = {Vertical and horizontal transmission drive bacterial invasion.}, journal = {Molecular ecology}, volume = {20}, number = {17}, pages = {3496-3498}, doi = {10.1111/j.1365-294X.2011.05194.x}, pmid = {21884292}, issn = {1365-294X}, mesh = {Animals ; Female ; *Genes, Bacterial ; *Host-Pathogen Interactions ; Male ; Wasps/*genetics ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {A huge variety of Arthropod species is infected with endosymbiotic Wolbachia bacteria that manipulate their host's reproduction to invade populations. In addition to vertical transmission from mother to offspring through the egg cytoplasm, it has been demonstrated through phylogenetic analyses and natural transfer experiments that horizontal transmission of Wolbachia (i.e. contagion) can occur between Arthropod hosts. More recently, factors influencing horizontal transfer have also been explored. While it is clear that horizontal transmission between species plays a major role in the evolutionary history of Wolbachia infections among insects, its role in the spread of a new infection through a host population, notably through within-species transfers, remained unknown. In this issue of Molecular Ecology, Kraaijeveld et al. (2011) present the first evidence that horizontal transmission played a key role in the early spread of parthenogenesis-inducing Wolbachia through the parasitoid wasp Leptopilina clavipes. To support their finding, the authors studied genetic variation in three types of markers, including host nuclear DNA, mitochondrial DNA and Wolbachia DNA. Specifically, they examined potential associations between their diversity patterns. No diversity was detected in Wolbachia genes, indicating that a single Wolbachia strain must have infected and spread through L. clavipes. In addition, a correlation between substantial variation in mitochondrial and nuclear genotypes suggested that horizontal transmission played an important role in the current clonal genetic variation in this wasp. Such horizontal transmission could be facilitated by a specific host ecology (e.g. parasitoid wasps sharing the same host resource) and potentially impact co-evolution between host and symbiont.}, } @article {pmid21880685, year = {2011}, author = {Watts, G}, title = {Bacterium blocks transmission of dengue by mosquitoes.}, journal = {BMJ (Clinical research ed.)}, volume = {343}, number = {}, pages = {d5545}, doi = {10.1136/bmj.d5545}, pmid = {21880685}, issn = {1756-1833}, mesh = {Aedes/microbiology/*physiology ; Animals ; Dengue/prevention & control/*transmission ; *Dengue Virus ; Insect Vectors/microbiology/*physiology ; Wolbachia/*physiology ; }, } @article {pmid21870980, year = {2011}, author = {Mitsuhashi, W and Ikeda, H and Muraji, M}, title = {Fifty-year trend towards suppression of Wolbachia-induced male-killing by its butterfly host, Hypolimnas bolina.}, journal = {Journal of insect science (Online)}, volume = {11}, number = {}, pages = {92}, pmid = {21870980}, issn = {1536-2442}, mesh = {Animals ; Biological Evolution ; Butterflies/genetics/immunology/*microbiology ; Female ; *Host-Pathogen Interactions ; Male ; Reproduction ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Some intracellular symbionts of arthropods induce a variety of reproductive alterations in their hosts, and the alterations tend to spread easily within the host populations. A few cases involving the spread of alteration-inducing Wolbachia bacteria in natural populations with time have been reported, but the investigations on the increasing trend in counteracting the bacterial effect on hosts in natural populations (i.e., increased resistance in hosts against the alterations) have been limited. In the present study, the prevalence of an alteration, killing of male Hypolimnas bolina (L.) (Lepidoptera: Nymphalidae) butterflies by their inherited Wolbachia strain in the wild in Japan, was surveyed over a continuous 50-year period, which is far longer than ever before analyzed in studies of dynamics between reproductive alteration-inducing symbionts and their host arthropods. Thus, the results in this study provide the first instance of a long-term trend involving a change in reproductive alteration; and it strongly suggests a change in the opposite direction (i.e., suppression of male-killing) in natural populations. This change in the current combination of the Wolbachia and butterflies appears to be dependent upon the host taxon (race).}, } @article {pmid21866160, year = {2011}, author = {Hoffmann, AA and Montgomery, BL and Popovici, J and Iturbe-Ormaetxe, I and Johnson, PH and Muzzi, F and Greenfield, M and Durkan, M and Leong, YS and Dong, Y and Cook, H and Axford, J and Callahan, AG and Kenny, N and Omodei, C and McGraw, EA and Ryan, PA and Ritchie, SA and Turelli, M and O'Neill, SL}, title = {Successful establishment of Wolbachia in Aedes populations to suppress dengue transmission.}, journal = {Nature}, volume = {476}, number = {7361}, pages = {454-457}, pmid = {21866160}, issn = {1476-4687}, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Dengue/microbiology/*prevention & control/*transmission/virology ; Dengue Virus/isolation & purification/*physiology ; Drosophila melanogaster/microbiology ; Female ; Humans ; Insect Vectors/microbiology/physiology/virology ; Male ; Pest Control, Biological/*methods ; Queensland ; Time Factors ; Wolbachia/isolation & purification/*physiology ; }, abstract = {Genetic manipulations of insect populations for pest control have been advocated for some time, but there are few cases where manipulated individuals have been released in the field and no cases where they have successfully invaded target populations. Population transformation using the intracellular bacterium Wolbachia is particularly attractive because this maternally-inherited agent provides a powerful mechanism to invade natural populations through cytoplasmic incompatibility. When Wolbachia are introduced into mosquitoes, they interfere with pathogen transmission and influence key life history traits such as lifespan. Here we describe how the wMel Wolbachia infection, introduced into the dengue vector Aedes aegypti from Drosophila melanogaster, successfully invaded two natural A. aegypti populations in Australia, reaching near-fixation in a few months following releases of wMel-infected A. aegypti adults. Models with plausible parameter values indicate that Wolbachia-infected mosquitoes suffered relatively small fitness costs, leading to an unstable equilibrium frequency <30% that must be exceeded for invasion. These findings demonstrate that Wolbachia-based strategies can be deployed as a practical approach to dengue suppression with potential for area-wide implementation.}, } @article {pmid21866159, year = {2011}, author = {Walker, T and Johnson, PH and Moreira, LA and Iturbe-Ormaetxe, I and Frentiu, FD and McMeniman, CJ and Leong, YS and Dong, Y and Axford, J and Kriesner, P and Lloyd, AL and Ritchie, SA and O'Neill, SL and Hoffmann, AA}, title = {The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations.}, journal = {Nature}, volume = {476}, number = {7361}, pages = {450-453}, pmid = {21866159}, issn = {1476-4687}, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Dengue/*prevention & control/transmission/virology ; Dengue Virus/classification/isolation & purification/*physiology ; Female ; Genetic Fitness ; Humans ; Insect Vectors/microbiology/physiology/virology ; Male ; Pest Control, Biological/*methods ; Reproduction/physiology ; Saliva/virology ; Wolbachia/*classification/*physiology ; }, abstract = {Dengue fever is the most important mosquito-borne viral disease of humans with more than 50 million cases estimated annually in more than 100 countries. Disturbingly, the geographic range of dengue is currently expanding and the severity of outbreaks is increasing. Control options for dengue are very limited and currently focus on reducing population abundance of the major mosquito vector, Aedes aegypti. These strategies are failing to reduce dengue incidence in tropical communities and there is an urgent need for effective alternatives. It has been proposed that endosymbiotic bacterial Wolbachia infections of insects might be used in novel strategies for dengue control. For example, the wMelPop-CLA Wolbachia strain reduces the lifespan of adult A. aegypti mosquitoes in stably transinfected lines. This life-shortening phenotype was predicted to reduce the potential for dengue transmission. The recent discovery that several Wolbachia infections, including wMelPop-CLA, can also directly influence the susceptibility of insects to infection with a range of insect and human pathogens has markedly changed the potential for Wolbachia infections to control human diseases. Here we describe the successful transinfection of A. aegypti with the avirulent wMel strain of Wolbachia, which induces the reproductive phenotype cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, providing optimal phenotypic effects for invasion. Under semi-field conditions, the wMel strain increased from an initial starting frequency of 0.65 to near fixation within a few generations, invading A. aegypti populations at an accelerated rate relative to trials with the wMelPop-CLA strain. We also show that wMel and wMelPop-CLA strains block transmission of dengue serotype 2 (DENV-2) in A. aegypti, forming the basis of a practical approach to dengue suppression.}, } @article {pmid21866151, year = {2011}, author = {Rasgon, JL}, title = {Dengue fever: Mosquitoes attacked from within.}, journal = {Nature}, volume = {476}, number = {7361}, pages = {407-408}, pmid = {21866151}, issn = {1476-4687}, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Dengue/microbiology/prevention & control/*transmission/virology ; Dengue Virus/*physiology ; Drosophila melanogaster/microbiology ; Humans ; Insect Vectors/microbiology/*physiology/virology ; Pest Control, Biological/*methods ; Queensland ; Time Factors ; Wolbachia/*physiology ; }, } @article {pmid21835008, year = {2011}, author = {Melnikov, A and Galinsky, K and Rogov, P and Fennell, T and Van Tyne, D and Russ, C and Daniels, R and Barnes, KG and Bochicchio, J and Ndiaye, D and Sene, PD and Wirth, DF and Nusbaum, C and Volkman, SK and Birren, BW and Gnirke, A and Neafsey, DE}, title = {Hybrid selection for sequencing pathogen genomes from clinical samples.}, journal = {Genome biology}, volume = {12}, number = {8}, pages = {R73}, pmid = {21835008}, issn = {1474-760X}, support = {T32 AI007638/AI/NIAID NIH HHS/United States ; HG03067-05/HG/NHGRI NIH HHS/United States ; HHSN27220090018C//PHS HHS/United States ; }, mesh = {Chromosome Mapping/methods ; DNA, Protozoan/*genetics ; *Genome, Protozoan ; Humans ; Malaria, Falciparum/*parasitology ; Nucleic Acid Hybridization/methods ; Plasmodium falciparum/*genetics ; Sequence Analysis, DNA/*methods ; }, abstract = {We have adapted a solution hybrid selection protocol to enrich pathogen DNA in clinical samples dominated by human genetic material. Using mock mixtures of human and Plasmodium falciparum malaria parasite DNA as well as clinical samples from infected patients, we demonstrate an average of approximately 40-fold enrichment of parasite DNA after hybrid selection. This approach will enable efficient genome sequencing of pathogens from clinical samples, as well as sequencing of endosymbiotic organisms such as Wolbachia that live inside diverse metazoan phyla.}, } @article {pmid21829750, year = {2011}, author = {Chambers, EW and Hapairai, L and Peel, BA and Bossin, H and Dobson, SL}, title = {Male mating competitiveness of a Wolbachia-introgressed Aedes polynesiensis strain under semi-field conditions.}, journal = {PLoS neglected tropical diseases}, volume = {5}, number = {8}, pages = {e1271}, pmid = {21829750}, issn = {1935-2735}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; AI-067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*physiology ; Animals ; Competitive Behavior/*physiology ; Eggs ; Elephantiasis, Filarial/*prevention & control/transmission ; Female ; Insect Vectors/microbiology/physiology ; Male ; Mosquito Control/*methods ; Sexual Behavior, Animal/*physiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Lymphatic filariasis (LF), a global public health problem affecting approximately 120 million people worldwide, is a leading cause of disability in the developing world including the South Pacific. Despite decades of ongoing mass drug administration (MDA) in the region, some island nations have not yet achieved the threshold levels of microfilaremia established by the World Health Organization for eliminating transmission. Previously, the generation of a novel Aedes polynesiensis strain (CP) infected with an exogenous type of Wolbachia has been described. The CP mosquito is cytoplasmically incompatible (i.e., effectively sterile) when mated with wildtype mosquitoes, and a strategy was proposed for the control of A. polynesiensis populations by repeated, inundative releases of CP males to disrupt fertility of wild females. Such a strategy could lead to suppression of the vector population and subsequently lead to a reduction in the transmission of filarial worms.

CP males and F1 male offspring from wild-caught A. polynesiensis females exhibit near equal mating competitiveness with F1 females under semi-field conditions.

CONCLUSIONS/SIGNIFICANCE: While laboratory experiments are important, prior projects have demonstrated the need for additional testing under semi-field conditions in order to recognize problems before field implementation. The results reported here from semi-field experiments encourage forward progression toward small-scale field releases.}, } @article {pmid21829485, year = {2011}, author = {Cho, KO and Kim, GW and Lee, OK}, title = {Wolbachia bacteria reside in host Golgi-related vesicles whose position is regulated by polarity proteins.}, journal = {PloS one}, volume = {6}, number = {7}, pages = {e22703}, pmid = {21829485}, issn = {1932-6203}, mesh = {Animals ; *Cell Polarity ; Drosophila Proteins/*metabolism ; Drosophila melanogaster/embryology/metabolism/*microbiology ; Embryo, Nonmammalian/metabolism/*microbiology ; Female ; Golgi Apparatus/metabolism/*microbiology ; Immunoenzyme Techniques ; Male ; Microtubules/metabolism/microbiology ; Secretory Vesicles/metabolism/microbiology ; *Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia pipientis are intracellular symbiotic bacteria extremely common in various organisms including Drosophila melanogaster, and are known for their ability to induce changes in host reproduction. These bacteria are present in astral microtubule-associated vesicular structures in host cytoplasm, but little is known about the identity of these vesicles. We report here that Wolbachia are restricted only to a group of Golgi-related vesicles concentrated near the site of membrane biogenesis and minus-ends of microtubules. The Wolbachia vesicles were significantly mislocalized in mutant embryos defective in cell/planar polarity genes suggesting that cell/tissue polarity genes are required for apical localization of these Golgi-related vesicles. Furthermore, two of the polarity proteins, Van Gogh/Strabismus and Scribble, appeared to be present in these Golgi-related vesicles. Thus, establishment of polarity may be closely linked to the precise insertion of Golgi vesicles into the new membrane addition site.}, } @article {pmid21828990, year = {2011}, author = {Fenton, A and Johnson, KN and Brownlie, JC and Hurst, GD}, title = {Solving the Wolbachia paradox: modeling the tripartite interaction between host, Wolbachia, and a natural enemy.}, journal = {The American naturalist}, volume = {178}, number = {3}, pages = {333-342}, doi = {10.1086/661247}, pmid = {21828990}, issn = {1537-5323}, mesh = {Animals ; Arthropods/immunology/*microbiology ; Female ; Host-Pathogen Interactions/*immunology ; Immune Tolerance ; Immunity, Innate ; *Models, Biological ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia is one of the most common symbionts of arthropods. Its establishment requires lateral transfer to and successful transmission within novel host species. However, Wolbachia performs poorly when introduced into new host species, and models predict that Wolbachia should seldom be able to establish from low initial frequencies. Recently, various symbionts, including Wolbachia, have been shown to protect their hosts from natural enemies. Hence, Wolbachia invasion may be facilitated by the dynamic interaction between it, its host, and a natural enemy. We model such an interaction whereby Wolbachia induces either complete resistance, partial resistance, or tolerance to a host-specific pathogen and also induces the common manipulation phenotype of cytoplasmic incompatibility (CI). We show that the presence of the pathogen greatly facilitates Wolbachia invasion from rare and widens the parameter space in which "imperfect" Wolbachia strains can invade. Furthermore, positive frequency-dependent selection through CI can drive Wolbachia to very high frequencies, potentially excluding the pathogen. These results may explain a poorly understood aspect of Wolbachia biology: it is widespread, despite performing poorly after transfer to new host species. They also support the intriguing possibility that Wolbachia strains that encode both CI and natural-enemy resistance could potentially rid insects, including human disease vectors, of important pathogens.}, } @article {pmid21828986, year = {2011}, author = {Barton, NH and Turelli, M}, title = {Spatial waves of advance with bistable dynamics: cytoplasmic and genetic analogues of Allee effects.}, journal = {The American naturalist}, volume = {178}, number = {3}, pages = {E48-75}, doi = {10.1086/661246}, pmid = {21828986}, issn = {1537-5323}, mesh = {Animals ; Biological Evolution ; Gene Flow ; Insecta/*genetics/microbiology ; *Models, Genetic ; Population Dynamics ; Wolbachia ; }, abstract = {Unlike unconditionally advantageous "Fisherian" variants that tend to spread throughout a species range once introduced anywhere, "bistable" variants, such as chromosome translocations, have two alternative stable frequencies, absence and (near) fixation. Analogous to populations with Allee effects, bistable variants tend to increase locally only once they become sufficiently common, and their spread depends on their rate of increase averaged over all frequencies. Several proposed manipulations of insect populations, such as using Wolbachia or "engineered underdominance" to suppress vector-borne diseases, produce bistable rather than Fisherian dynamics. We synthesize and extend theoretical analyses concerning three features of their spatial behavior: rate of spread, conditions to initiate spread from a localized introduction, and wave stopping caused by variation in population densities or dispersal rates. Unlike Fisherian variants, bistable variants tend to spread spatially only for particular parameter combinations and initial conditions. Wave initiation requires introduction over an extended region, while subsequent spatial spread is slower than for Fisherian waves and can easily be halted by local spatial inhomogeneities. We present several new results, including robust sufficient conditions to initiate (and stop) spread, using a one-parameter cubic approximation applicable to several models. The results have both basic and applied implications.}, } @article {pmid21827871, year = {2011}, author = {Shiny, C and Krushna, NS and Babu, S and Elango, S and Manokaran, G and Narayanan, RB}, title = {Recombinant Wolbachia heat shock protein 60 (HSP60) mediated immune responses in patients with lymphatic filariasis.}, journal = {Microbes and infection}, volume = {13}, number = {14-15}, pages = {1221-1231}, pmid = {21827871}, issn = {1769-714X}, support = {Y99 AI999999/AI/NIAID NIH HHS/United States ; Z99 AI999999/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Adolescent ; Adult ; Animals ; Bacterial Proteins/genetics/*immunology/pharmacology ; Brugia malayi/immunology/*microbiology ; CTLA-4 Antigen/immunology ; Cells, Cultured ; Chaperonin 60/genetics/*immunology/pharmacology ; Child ; Elephantiasis, Filarial/*immunology/parasitology ; Female ; Humans ; Immunomodulation/drug effects ; Interferon-gamma/biosynthesis/immunology ; Interleukin-10/biosynthesis/immunology ; Interleukin-2 Receptor alpha Subunit/immunology ; Lymphocyte Activation/drug effects ; Male ; Recombinant Proteins/genetics/*immunology/pharmacology ; Symbiosis ; T-Lymphocytes, Regulatory/drug effects/*immunology ; Transforming Growth Factor beta/biosynthesis/immunology ; Wolbachia/chemistry/physiology ; }, abstract = {Wolbachia, an endosymbiont present in filarial nematodes, have been implicated in a variety of roles, including the worm development and survival. Elucidation of the role of Wolbachia in filarial nematode biology and pathogenesis has become the focus of many studies and its contribution to parasite survival or immune response is still unclear. Recombinant Wolbachia HSP60 decreases T cell activation and lymphoproliferation in filarial infected people compared to endemic controls as observed by the assessment of T cell activation markers and cytokine responses in the peripheral blood mononuclear cells. Reduced T cell activation may be linked to T regulatory cell activity since it is associated with increased expression of CTLA4 and CD25 on CD4(+) T cells in filarial infected group upon stimulation with recombinant Wolbachia HSP60. In addition, elevated interleukin-10 and TGF-β cytokines corroborate the reduced CD4(+) T cell activation and interferon-γ observed upon recombinant Wolbachia HSP60 stimulation in filarial patients. Hence, these findings indicate that Wolbachia HSP60 may also contribute to the immune modulation seen in filarial patients.}, } @article {pmid21814492, year = {2011}, author = {Sinkins, SP}, title = {Invertebrate post-segregation distorters: a new embryo-killing gene.}, journal = {PLoS biology}, volume = {9}, number = {7}, pages = {e1001114}, pmid = {21814492}, issn = {1545-7885}, support = {079059//Wellcome Trust/United Kingdom ; 095121//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Bacteroidetes/physiology ; *Chromosome Segregation ; Female ; Heredity ; Male ; Reproduction ; Tribolium/embryology/*genetics/microbiology ; Wolbachia/physiology ; }, abstract = {Cytoplasmic incompatibility induced by inherited intracellular bacteria of arthropods, and Medea elements found in flour beetles, are both forms of postsegregation distortion involving the killing of embryos in order to increase the ratio of progeny that inherit them. The recently described peel-zeel element of Caenorhabditis elegans also uses this mechanism; like Medea the genes responsible are in the nuclear genome but it shares a paternal mode of action with the bacteria. The peel-1 gene has now been shown to encode a potent toxin that is delivered by sperm, and rescued by zygotic transcription of the linked zeel-1. The predominance of self-fertilization in C. elegans has produced an unusual distribution pattern for a selfish genetic element; further population and functional studies will shed light on its evolution. The element might also have potential for use in disease control.}, } @article {pmid21799900, year = {2011}, author = {Carrington, LB and Lipkowitz, JR and Hoffmann, AA and Turelli, M}, title = {A re-examination of Wolbachia-induced cytoplasmic incompatibility in California Drosophila simulans.}, journal = {PloS one}, volume = {6}, number = {7}, pages = {e22565}, pmid = {21799900}, issn = {1932-6203}, mesh = {Animals ; California ; Cytoplasm/*microbiology ; Drosophila/*cytology/genetics/*microbiology/physiology ; Female ; Fertility ; Gram-Negative Bacterial Infections/transmission ; Hybridization, Genetic ; Infertility/*microbiology/pathology ; Male ; Mothers ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: In California Drosophila simulans, the maternally inherited Riverside strain Wolbachia infection (wRi) provides a paradigm for rapid spread of Wolbachia in nature and rapid evolutionary change. wRi induces cytoplasmic incompatibility (CI), where crosses between infected males and uninfected females produce reduced egg-hatch. The three parameters governing wRi infection-frequency dynamics quantify: the fidelity of maternal transmission, the level of cytoplasmic incompatibility, and the relative fecundity of infected females. We last estimated these parameters in nature in 1993. Here we provide new estimates, under both field and laboratory conditions. Five years ago, we found that wRi had apparently evolved over 15 years to enhance the fecundity of infected females; here we examine whether CI intensity has also evolved.

New estimates using wild-caught flies indicate that the three key parameters have remained relatively stable since the early 1990s. As predicted by our three-parameter model using field-estimated parameter values, population infection frequencies remain about 93%. Despite this relative stability, laboratory data based on reciprocal crosses and introgression suggest that wRi may have evolved to produce less intense CI (i.e., higher egg hatch from incompatible crosses). In contrast, we find no evidence that D. simulans has evolved to lower the susceptibility of uninfected females to CI.

CONCLUSIONS/SIGNIFICANCE: Evolution of wRi that reduces CI is consistent with counterintuitive theoretical predictions that within-population selection on CI-causing Wolbachia does not act to increase CI. Within taxa, CI is likely to evolve mainly via pleiotropic effects associated with the primary targets of selection on Wolbachia, i.e., host fecundity and transmission fidelity. Despite continuous, strong selection, D. simulans has not evolved appreciably to suppress CI. Our data demonstrate a lack of standing genetic variation for CI resistance in the host.}, } @article {pmid21792226, year = {2012}, author = {Raychoudhury, R and Werren, JH}, title = {Host genotype changes bidirectional to unidirectional cytoplasmic incompatibility in Nasonia longicornis.}, journal = {Heredity}, volume = {108}, number = {2}, pages = {105-114}, pmid = {21792226}, issn = {1365-2540}, mesh = {Animals ; Cytoplasm/genetics/*microbiology/physiology ; Female ; Genotype ; Host Specificity ; Male ; Reproduction ; Symbiosis ; Wasps/*genetics/microbiology/physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are the most abundant maternally inherited endosymbionts of insects and cause various reproductive alterations in their hosts. One such manipulation is cytoplasmic incompatibility (CI), which is a sperm-egg incompatibility typically resulting in zygotic death. Nasonia longicornis (Hymenoptera: Pteromalidae) has an A supergroup and two closely related B supergroup Wolbachia infections. The B supergroup bacteria co-diverged in this host genus. Both triple (wNlonAwNlonB1wNlonB2) and double infections (wNlonAwNlonB1, wNlonAwNlonB2) have been obtained from the field. In the present study, CI was determined among the three Wolbachia types in different host genetic backgrounds. Results show that host genetic background determines whether bidirectional CI or unidirectional CI occurs between the two closely related B group Wolbachia. Results show that the wNlonB1-infected males are bidirectionally incompatible with wNlonB2 in their 'native' nuclear genetic background, whereas wNlonB1 males are compatible with wNlonB2 in two other N. longicornis genetic backgrounds, resulting in unidirectional CI. In contrast, wNlonB2-infected males are incompatible with wNlonB1 females in all three host genetic backgrounds. These changes in incompatibility are not due to the loss of the bacteria. We hypothesize that a repressor gene for sperm modification by wNlonB1 is segregating in N. longicornis populations. The relevance of these findings to the potential role of Wolbachia in host-reproductive divergence and speciation is discussed.}, } @article {pmid21789233, year = {2011}, author = {Arthofer, W and Riegler, M and Schuler, H and Schneider, D and Moder, K and Miller, WJ and Stauffer, C}, title = {Allele intersection analysis: a novel tool for multi locus sequence assignment in multiply infected hosts.}, journal = {PloS one}, volume = {6}, number = {7}, pages = {e22198}, pmid = {21789233}, issn = {1932-6203}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {*Alleles ; Animals ; Bacterial Typing Techniques ; Base Sequence ; Bias ; Ceratitis capitata/microbiology ; Computer Simulation ; Host-Pathogen Interactions/*genetics ; Multilocus Sequence Typing/*methods ; Species Specificity ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia are wide-spread, endogenous α-Proteobacteria of arthropods and filarial nematodes. 15-75% of all insect species are infected with these endosymbionts that alter their host's reproduction to facilitate their spread. In recent years, many insect species infected with multiple Wolbachia strains have been identified. As the endosymbionts are not cultivable outside living cells, strain typing relies on molecular methods. A Multi Locus Sequence Typing (MLST) system was established for standardizing Wolbachia strain identification. However, MLST requires hosts to harbour individual and not multiple strains of supergroups without recombination. This study revisits the applicability of the current MLST protocols and introduces Allele Intersection Analysis (AIA) as a novel approach. AIA utilizes natural variations in infection patterns and allows correct strain assignment of MLST alleles in multiply infected host species without the need of artificial strain segregation. AIA identifies pairs of multiply infected individuals that share Wolbachia and differ in only one strain. In such pairs, the shared MLST sequences can be used to assign alleles to distinct strains. Furthermore, AIA is a powerful tool to detect recombination events. The underlying principle of AIA may easily be adopted for MLST approaches in other uncultivable bacterial genera that occur as multiple strain infections and the concept may find application in metagenomic high-throughput parallel sequencing projects.}, } @article {pmid21786068, year = {2012}, author = {Shiny, C and Krushna, NS and Haripriya, K and Babu, S and Elango, S and Manokaran, G and Narayanan, RB}, title = {Recombinant Wolbachia surface protein (WSP)-induced T cell responses in Wuchereria bancrofti infections.}, journal = {Parasitology research}, volume = {110}, number = {2}, pages = {787-797}, pmid = {21786068}, issn = {1432-1955}, support = {Z99 AI999999//Intramural NIH HHS/United States ; }, mesh = {Animals ; Antigens, CD/analysis ; Antigens, Differentiation, T-Lymphocyte/analysis ; Bacterial Outer Membrane Proteins/*immunology ; CTLA-4 Antigen/analysis ; Cell Proliferation ; Cytokines/metabolism ; Elephantiasis, Filarial/*immunology ; Humans ; Interleukin-2 Receptor alpha Subunit/analysis ; Interleukin-7 Receptor alpha Subunit/analysis ; L-Selectin/analysis ; Lectins, C-Type/analysis ; Leukocytes, Mononuclear/chemistry/immunology ; Membrane Proteins/*immunology ; T-Lymphocytes/chemistry/*immunology ; T-Lymphocytes, Regulatory/chemistry/immunology ; Wolbachia/*immunology ; Wuchereria bancrofti/*microbiology/pathogenicity ; }, abstract = {Human lymphatic filariasis is a debilitating parasitic disease characterized by downregulation of the host's immune response in asymptomatic carriers along with profound hyperreactivity in chronic patients apart from putatively immune endemic normals. The endosymbiont Wolbachia, a bacterium of filarial nematodes has received much attention as possible chemotherapeutic target and its involvement in disease pathogenesis. The role of recombinant Wolbachia surface protein (rWSP), one of the most abundantly expressed proteins of the endosymbiont, in modulating cell-mediated immune responses in patients harboring Wuchereria bancrofti infections was evaluated in the current study. rWSP-induced lymphoproliferation with peripheral blood mononuclear cells suggested an impaired proliferative response in asymptomatic microfilaremic (MF) and symptomatic chronic pathology (CP) patients compared to endemic normals (EN). This was further supported by a significantly diminished expression of CD69 along with elevated levels of CD127 and CD62L in filarial patients (MF and CP) compared to EN. Further, rWSP induced the expression of regulatory T cell markers CTLA-4 and CD25 along with suppressor cytokines IL-10 and TGF-β in MF and CP patients compared to EN. However, the rWSP-stimulated expression of IFN-γ was diminished significantly in filarial patients compared to endemic normals. Thus, these findings suggest that WSP may also contribute to the suppression of immune responses seen in filarial patients.}, } @article {pmid21782817, year = {2011}, author = {Melnikow, E and Xu, S and Liu, J and Li, L and Oksov, Y and Ghedin, E and Unnasch, TR and Lustigman, S}, title = {Interaction of a Wolbachia WSP-like protein with a nuclear-encoded protein of Brugia malayi.}, journal = {International journal for parasitology}, volume = {41}, number = {10}, pages = {1053-1061}, pmid = {21782817}, issn = {1879-0135}, support = {R01 AI072465/AI/NIAID NIH HHS/United States ; R01 AI072465-03/AI/NIAID NIH HHS/United States ; AI072465/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/*metabolism ; Brugia malayi/genetics/*metabolism/*microbiology ; Enzyme-Linked Immunosorbent Assay ; Female ; *Host-Parasite Interactions ; Mice ; Microscopy ; Protein Binding ; *Protein Interaction Mapping ; Wolbachia/genetics/*metabolism ; }, abstract = {The Brugia malayi endosymbiont Wolbachia has recently been shown to be essential for its host's survival and development. However, relatively little is known about Wolbachia proteins that interact with the filarial host and which might be important in maintaining the obligate symbiotic relationship. The Wolbachia surface proteins (WSPs) are members of the outer membrane protein family and we hypothesise that they might be involved in the Wolbachia-Brugia symbiotic relationship. Notably, immunolocalisation studies of two WSP members, WSP-0432 and WSP-0284 in B. malayi female adult worms showed that the corresponding proteins are not only present on the surface of Wolbachia but also in the host tissues, with WSP-0284 more abundant in the cuticle, hypodermis and the nuclei within the embryos. These results confirmed that WSPs might be secreted by Wolbachia into the worm's tissue. Our present studies focus on the potential involvement of WSP-0284 in the symbiotic relationship of Wolbachia with its filarial host. We show that WSP-0284 binds specifically to B. malayi crude protein extracts. Furthermore, a fragment of the hypothetical B. malayi protein (Bm1_46455) was found to bind WSP-0284 by panning of a B. malayi cDNA library. The interaction of WSP-0284 and this protein was further confirmed by ELISA and pull-down assays. Localisation by immunoelectron microscopy within Wolbachia cells as well as in the worm's tissues, cuticle and nuclei within embryos established that both proteins are present in similar locations within the parasite and the bacteria. Identifying such specific interactions between B. malayi and Wolbachia proteins should lead to a better understanding of the molecular basis of the filarial nematode and Wolbachia symbiosis.}, } @article {pmid21776894, year = {2011}, author = {Guerrero, R and Bain, O and Martin, C and Barbuto, M}, title = {A new species of Litomosoides (Nematoda: Onchocercidae), parasite of Nectomys palmipes (Rodentia: Cricetidae: Sigmodontinae) from Venezuela: description, molecular evidence, Wolbachia pipientis screening.}, journal = {Folia parasitologica}, volume = {58}, number = {2}, pages = {149-156}, doi = {10.14411/fp.2011.014}, pmid = {21776894}, issn = {0015-5683}, mesh = {Animals ; DNA, Helminth/chemistry/genetics ; Female ; Filariasis/parasitology/*veterinary ; Filarioidea/anatomy & histology/*classification/*isolation & purification ; Genes, Helminth/genetics ; Genes, rRNA/genetics ; Male ; Polymerase Chain Reaction/veterinary ; Rodent Diseases/*parasitology ; Sequence Analysis, DNA/veterinary ; Sigmodontinae/*parasitology ; Venezuela ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Abstract: The onchocercid filaria Litomosoides taylori sp. n. is described from the sigmodontine cricetid Nectomys palmipes Allen et Chapman in northeast Venezuela. A voucher specimen of the new species was used for molecular analysis of the coxI and 12S rDNA genes, and screened for the presence of the endobacterium Wolbachia pipientis. Litomosoides taylori belongs to the "sigmodontis group" of Litomosoides and a combination of characters can be used to distinguish it from the remaining 18 species forming this group. Among the five Nectomys species, all living near running water, N. squamipes also harbours Litomosoides species, L. khonae in Brazil and L. navonae in Argentina. These three Litomosoides species of the "sigmodontis group" do not share any particular characters. Gene sequences of L. taylori differ from those of the five Litomosoides species available, the three of the "carinii group" being the most distant. The new species harbours W pipientis, which is concurrent with the great majority of Litomosoides species screened to date.}, } @article {pmid21745252, year = {2011}, author = {Kodandaramaiah, U and Weingartner, E and Janz, N and Dalén, L and Nylin, S}, title = {Population structure in relation to host-plant ecology and Wolbachia infestation in the comma butterfly.}, journal = {Journal of evolutionary biology}, volume = {24}, number = {10}, pages = {2173-2185}, doi = {10.1111/j.1420-9101.2011.02352.x}, pmid = {21745252}, issn = {1420-9101}, mesh = {Animals ; Butterflies/genetics/*microbiology ; DNA, Mitochondrial/chemistry ; Female ; Male ; Microsatellite Repeats ; Phylogeography ; Polymorphism, Genetic ; Population Dynamics ; Sequence Analysis, DNA ; Wolbachia/*physiology ; }, abstract = {Experimental work on Polygonia c-album, a temperate polyphagous butterfly species, has shown that Swedish, Belgian, Norwegian and Estonian females are generalists with respect to host-plant preference, whereas females from UK and Spain are specialized on Urticaceae. Female preference is known to have a strong genetic component. We test whether the specialist and generalist populations form respective genetic clusters using data from mitochondrial sequences and 10 microsatellite loci. Results do not support this hypothesis, suggesting that the specialist and generalist traits have evolved more than once independently. Mitochondrial DNA variation suggests a rapid expansion scenario, with a single widespread haplotype occurring in high frequency, whereas microsatellite data indicate strong differentiation of the Moroccan population. Based on a comparison of polymorphism in the mitochondrial data and sequences from a nuclear gene, we show that the diversity in the former is significantly less than that expected under neutral evolution. Furthermore, we found that almost all butterfly samples were infected with a single strain of Wolbachia, a maternally inherited bacterium. We reason that indirect selection on the mitochondrial genome mediated by a recent sweep of Wolbachia infection has depleted variability in the mitochondrial sequences. We also surmise that P. c-album could have expanded out of a single glacial refugium and colonized Morocco recently.}, } @article {pmid21745250, year = {2011}, author = {Elnagdy, S and Majerus, ME and Handley, LJ}, title = {The value of an egg: resource reallocation in ladybirds (Coleoptera: Coccinellidae) infected with male-killing bacteria.}, journal = {Journal of evolutionary biology}, volume = {24}, number = {10}, pages = {2164-2172}, doi = {10.1111/j.1420-9101.2011.02346.x}, pmid = {21745250}, issn = {1420-9101}, mesh = {Animals ; Coleoptera/*microbiology ; Female ; Larva/microbiology/physiology ; Male ; Ovum ; Rickettsia/physiology ; Sex Factors ; Species Specificity ; Spiroplasma/physiology ; Wolbachia/physiology ; }, abstract = {Male-killing bacteria are thought to persist in host populations by vertical transmission and conferring direct and/or indirect fitness benefits to their hosts. Here, we test the role of indirect fitness benefits accrued from resource reallocation in species that engage in sibling egg cannibalism. We found that a single-egg meal significantly increased larval survival in 12 ladybird species, but the value of an egg (to survival) differed substantially between species. Next, we tested the impact of three male-killing bacteria on larval survival in one ladybird species, Adalia bipunctata. Spiroplasma reduced larval survival, whereas Wolbachia and Rickettsia had no effect. However, Spiroplasma-infected larvae showed the greatest response to a single-egg meal. The indirect fitness benefit obtained from a single egg is thus so large that even male-killing bacteria with direct fitness costs can persist in host populations. This study supports the hypothesis that fitness compensation via resource reallocation can explain male-killing bacteria persistence.}, } @article {pmid21739251, year = {2011}, author = {Guruprasad, NM and Mouton, L and Sumithra, and Puttaraju, HP}, title = {Survey of Wolbachia and its phage WO in the Uzifly Exorista sorbillans (Diptera: Tachinidae).}, journal = {Current microbiology}, volume = {63}, number = {3}, pages = {267-272}, pmid = {21739251}, issn = {1432-0991}, mesh = {Animals ; Bacterial Proteins/genetics ; Bacteriophages/*isolation & purification ; Bombyx/parasitology ; Capsid Proteins/genetics ; DNA, Bacterial/chemistry/genetics ; DNA, Viral/chemistry/genetics ; Diptera/*microbiology ; India ; *Lysogeny ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/*isolation & purification/*virology ; }, abstract = {Wolbachia are cytoplasmically inherited alpha-proteobacteria well known for inducing a variety of reproductive abnormalities in the diverse arthropod hosts they infect. Despite their obligate intracellular lifestyle which usually protects bacteria from phage infection, Wolbachia harbor a widespread temperate phage called WO. Evidences of horizontal phage transfers indicate that this phage could promote genetic exchanges between strains leading to evolutionary changes in the genomes of Wolbachia, and could be involved in the phenotypes these bacteria induced. In this study, we report the survey of Wolbachia and WO phage infections in 20 populations of the Uzifly Exorista sorbillans, a tachinid endoparasite of silkworm Bombyx mori, collected from different geographic regions of India. Previous studies demonstrated that Wolbachia is associated with positive reproductive fitness effects in this species. Polymerase chain reaction using the ftsZ gene encoding for a Wolbachia cell division protein and the orf7 capsid protein gene of the phage showed that all flies checked were infected by Wolbachia and its phage WO. Phylogenetic analyses based on the Wolbachia surface protein gene revealed 100% of double infections by the arthropod supergroups A and B. These results can serve as a valuable basis for understanding the evolution of Wolbachia bacteria and may provide information about the dynamics of Wolbachia-host associations. This knowledge could be exploited for the use of Wolbachia for effective control strategies of the Uzifly, a serious menace of the silkworm B. mori.}, } @article {pmid21734243, year = {2011}, author = {Tamarozzi, F and Halliday, A and Gentil, K and Hoerauf, A and Pearlman, E and Taylor, MJ}, title = {Onchocerciasis: the role of Wolbachia bacterial endosymbionts in parasite biology, disease pathogenesis, and treatment.}, journal = {Clinical microbiology reviews}, volume = {24}, number = {3}, pages = {459-468}, pmid = {21734243}, issn = {1098-6618}, mesh = {Animals ; Diethylcarbamazine/adverse effects/therapeutic use ; Filaricides/adverse effects/therapeutic use ; Humans ; Inflammation/chemically induced/microbiology ; Ivermectin/adverse effects/therapeutic use ; Onchocerca volvulus/*microbiology ; Onchocerciasis, Ocular/drug therapy/*microbiology/*parasitology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The discovery of Wolbachia intracellular bacteria within filarial nematodes, including Onchocerca volvulus, the causative agent of onchocerciasis or "river blindness," has delivered a paradigm shift in our understanding of the parasite's biology, to where we now know that the bacterial endosymbionts are essential for normal development of larvae and embryos and may support the long-term survival of adult worms. The apparent mutualistic dependency has also offered a novel approach to the treatment of onchocerciasis through the use of antibiotics to eliminate Wolbachia, delivering for the first time a treatment which has significant macrofilaricidal efficacy. Studies with other filarial nematode species have also highlighted a role for Wolbachia in transmission and infection of the mammalian host through a fascinating manipulation of mast cell-mediated vasodilation to enhance infectivity of vector-borne larvae. Wolbachia has also been identified as the principal driver of innate and adaptive Th1 inflammatory immunity, which can either contribute to disease pathogenesis or, with the Wolbachia-mediated recruitment of mast cells, enhance infectivity. The Wolbachia activation of innate inflammation also drives inflammatory adverse events in response to chemotherapy with either diethylcarbamazine (DEC) or ivermectin. In this review we summarize the experimental and field trial data which have uncovered the importance of Wolbachia symbiosis in onchocerciasis.}, } @article {pmid21731626, year = {2011}, author = {Ferri, E and Bain, O and Barbuto, M and Martin, C and Lo, N and Uni, S and Landmann, F and Baccei, SG and Guerrero, R and de Souza Lima, S and Bandi, C and Wanji, S and Diagne, M and Casiraghi, M}, title = {New insights into the evolution of Wolbachia infections in filarial nematodes inferred from a large range of screened species.}, journal = {PloS one}, volume = {6}, number = {6}, pages = {e20843}, pmid = {21731626}, issn = {1932-6203}, mesh = {Animals ; Base Sequence ; *Biological Evolution ; DNA, Ribosomal/genetics ; Female ; Filarioidea/cytology/*isolation & purification/*microbiology ; Fluorescence ; Gram-Negative Bacterial Infections/*genetics/*microbiology ; Host Specificity ; Host-Pathogen Interactions ; Phylogeny ; Polymerase Chain Reaction ; Propidium/metabolism ; Species Specificity ; Wolbachia/cytology/genetics/*physiology ; }, abstract = {BACKGROUND: Wolbachia are intriguing symbiotic endobacteria with a peculiar host range that includes arthropods and a single nematode family, the Onchocercidae encompassing agents of filariases. This raises the question of the origin of infection in filariae. Wolbachia infect the female germline and the hypodermis. Some evidences lead to the theory that Wolbachia act as mutualist and coevolved with filariae from one infection event: their removal sterilizes female filariae; all the specimens of a positive species are infected; Wolbachia are vertically inherited; a few species lost the symbiont. However, most data on Wolbachia and filaria relationships derive from studies on few species of Onchocercinae and Dirofilariinae, from mammals.

We investigated the Wolbachia distribution testing 35 filarial species, including 28 species and 7 genera and/or subgenera newly screened, using PCR, immunohistochemical staining, whole mount fluorescent analysis, and cocladogenesis analysis. (i) Among the newly screened Onchocercinae from mammals eight species harbour Wolbachia but for some of them, bacteria are absent in the hypodermis, or in variable density. (ii) Wolbachia are not detected in the pathological model Monanema martini and in 8, upon 9, species of Cercopithifilaria. (iii) Supergroup F Wolbachia is identified in two newly screened Mansonella species and in Cercopithifilaria japonica. (iv) Type F Wolbachia infect the intestinal cells and somatic female genital tract. (v) Among Oswaldofilariinae, Waltonellinae and Splendidofilariinae, from saurian, anuran and bird respectively, Wolbachia are not detected.

CONCLUSIONS/SIGNIFICANCE: The absence of Wolbachia in 63% of onchocercids, notably in the ancestral Oswaldofilariinae estimated 140 mya old, the diverse tissues or specimens distribution, and a recent lateral transfer in supergroup F Wolbachia, modify the current view on the role and evolution of the endosymbiont and their hosts. Further genomic analyses on some of the newly sampled species are welcomed to decipher the open questions.}, } @article {pmid21731302, year = {2011}, author = {Winthrop, KL and Furtado, JM and Silva, JC and Resnikoff, S and Lansingh, VC}, title = {River blindness: an old disease on the brink of elimination and control.}, journal = {Journal of global infectious diseases}, volume = {3}, number = {2}, pages = {151-155}, pmid = {21731302}, issn = {0974-8245}, abstract = {For decades, onchocerciasis (or river blindness) was one of the most common infectious causes of blindness in the world. Primarily an infection of Africa, with limited distribution in the new world, disease due to the nematode Onchocerca volvulus is rapidly diminishing as a result of large public health campaigns targeting at risk populations in Africa and the Americas. Existing and newly-developed treatment strategies offer the chance to eliminate onchocercal ocular morbidity in some parts of the world. This article reviews these treatment strategies, current clinical and epidemiologic aspects of onchocerciasis, and the next steps toward elimination.}, } @article {pmid21728798, year = {2011}, author = {Kondo, NI and Tuda, M and Toquenaga, Y and Lan, YC and Buranapanichpan, S and Horng, SB and Shimada, M and Fukatsu, T}, title = {Wolbachia infections in world populations of bean beetles (Coleoptera: Chrysomelidae: Bruchinae) infesting cultivated and wild legumes.}, journal = {Zoological science}, volume = {28}, number = {7}, pages = {501-508}, doi = {10.2108/zsj.28.501}, pmid = {21728798}, issn = {0289-0003}, mesh = {Animals ; Asia ; Coleoptera/*microbiology ; Crops, Agricultural/parasitology ; DNA, Bacterial/isolation & purification ; Fabaceae/*parasitology ; Middle East ; Phylogeny ; Uganda ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia endosymbionts are widespread among insects and other arthropods, often causing cytoplasmic incompatibility and other reproductive phenotypes in their hosts. Recently, possibilities of Wolbachia-mediated pest control and management have been proposed, and the bean beetles of the subfamily Bruchinae are known as serious pests of harvested and stored beans worldwide. Here we investigated Wolbachia infections in bean beetles from the world, representing seven genera, 20 species and 87 populations. Of 20 species examined, Wolbachia infections were detected in four species, Megabruchidius sophorae, Callosobruchus analis, C. latealbus and C. chinensis. Infection frequencies were partial in M. sophorae but perfect in the other species. In addition to C. chinensis described in the previous studies, C. latealbus was infected with two distinct Wolbachia strains. These Wolbachia strains from the bean beetles were phylogenetically not closely related to each other. Among world populations of C. chinensis, some Taiwanese populations on a wild leguminous plant, Rhynchosia minima, exhibited a peculiar Wolbachia infection pattern, suggesting the possibility that these populations comprise a distinct host race or a cryptic species.}, } @article {pmid21726632, year = {2011}, author = {Papafotiou, G and Oehler, S and Savakis, C and Bourtzis, K}, title = {Regulation of Wolbachia ankyrin domain encoding genes in Drosophila gonads.}, journal = {Research in microbiology}, volume = {162}, number = {8}, pages = {764-772}, doi = {10.1016/j.resmic.2011.06.012}, pmid = {21726632}, issn = {1769-7123}, mesh = {Animals ; Animals, Genetically Modified ; Ankyrin Repeat ; Bacterial Proteins/chemistry/genetics/metabolism ; Drosophila/*genetics/metabolism/*microbiology ; Female ; *Gene Expression Regulation ; Gonads/metabolism/microbiology ; Male ; Molecular Sequence Data ; Organ Specificity ; Wolbachia/chemistry/*genetics ; }, abstract = {The maternally inherited obligatory intracellular bacterium Wolbachia is a reproductive parasite of many insect species. Wolbachia evades the host immune system, uses the mitotic apparatus to ensure infection of daughter cells, migrates through the host to the gonads and causes reproductive phenotypes, most commonly cytoplasmic incompatibility (CI), i.e. incompatibility of sperm from infected males and eggs from uninfected females. Due to the interconnected facts that Wolbachia is not ex vivo culturable and that no established transformation system exists, virtually nothing is known about Wolbachia-host interactions at the macromolecular level. Intriguingly, the Wolbachia genome codes for an unusually high number of ankyrin repeat (ANK) proteins. ANKs mediate protein-protein interactions in many different contexts. More common in eukaryotes, they also occur in prokaryotes. Some intracellular pathogenic bacteria export ANK effector proteins to the host cytoplasm. This makes the Wolbachia ANK genes candidates for mediating interactions with host cells. We quantified expression of ANK genes of Wolbachia strain wMel in adult gonads and detected host sex-specific regulation of two wMel ANK genes in the gonads in two different backgrounds. Regulation was tissue-specific and independent of host background. We further analyzed expression of their homologues in strains wAu and wRi and found regulation only in wAu. Regulation was tissue-specific and there was no correlation between regulation of these genes and the ability of a strain to induce CI.}, } @article {pmid21722251, year = {2011}, author = {Hoerauf, A and Pfarr, K and Mand, S and Debrah, AY and Specht, S}, title = {Filariasis in Africa--treatment challenges and prospects.}, journal = {Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases}, volume = {17}, number = {7}, pages = {977-985}, doi = {10.1111/j.1469-0691.2011.03586.x}, pmid = {21722251}, issn = {1469-0691}, mesh = {Africa/epidemiology ; Albendazole/adverse effects/pharmacology/therapeutic use ; Animals ; Elephantiasis, Filarial/*drug therapy/*epidemiology ; Filaricides/pharmacology/therapeutic use ; Humans ; Ivermectin/adverse effects/pharmacology/therapeutic use ; Onchocerca volvulus/microbiology/pathogenicity/physiology ; Onchocerciasis/*drug therapy/*epidemiology ; Symbiosis ; Wolbachia/physiology ; Wuchereria bancrofti/microbiology/pathogenicity/physiology ; }, abstract = {Lymphatic filariasis (LF) and onchocerciasis are parasitic nematode infections that are responsible for a major disease burden in the African continent. Disease symptoms are induced by the immune reactions of the host, with lymphoedema and hydrocoele in LF, and dermatitis and ocular inflammation in onchocerciasis. Wuchereria bancrofti and Onchocerca volvulus, the species causing LF and onchocerciasis in Africa, live in mutual symbiosis with Wolbachia endobacteria, which cause a major part of the inflammation leading to symptoms and are antibiotic targets for treatment. The standard microfilaricidal drugs ivermectin and albendazole are used in mass drug administration programmes, with the aim of interrupting transmission, with a consequent reduction in the burden of infection and, in some situations, leading to regional elimination of LF and onchocerciasis. Co-endemicity of Loa loa with W. bancrofti or O. volvulus is an impediment to mass drug administration with ivermectin and albendazole, owing to the risk of encephalopathy being encountered upon administration of ivermectin. Research into new treatment options is exploring several improved delivery strategies for the classic drugs or new antibiotic treatment regimens for anti-wolbachial chemotherapy.}, } @article {pmid23508470, year = {2011}, author = {Dhamodharan, R and Hoti, S and Sivapragasam, G and Das, M}, title = {Cloning and sequence analysis of partial genomic DNA coding for HtrA-type serine protease of Wolbachia from human lymphatic filarial parasite, Wuchereria bancrofti.}, journal = {Tropical parasitology}, volume = {1}, number = {2}, pages = {76-82}, pmid = {23508470}, issn = {2229-5070}, abstract = {BACKGROUND: Periplasmic serine proteases of HtrA type of Wolbachia have been shown to play a role in the pathogenesis of filarial disease.

AIMS: This study was aimed to sequence Wb-HtrA serine protease and analyze its phylogenetic position by comparing with other filarial and non-filarial nematode homologs.

MATERIALS AND METHODS: Partial HtrA gene fragment was amplified from DNA isolated from periodic and sub-periodic Wuchereria bancrofti parasites collected from Pondicherry and Nicobar islands, respectively. The amplicons were sequenced, and sequence homology and phylogenetic relationship with other filarial and non-filarial nematodes were analyzed.

RESULTS: Partial orthologue of HtrA-type serine protease from Wolbachia of W. bancrofti was amplified, cloned and sequenced. The deduced amino acid sequence exhibited 87%, 81% and 74% identity with the homologous Wolbachia proteases identified from Brugia malayi, Onchocerca volvulus and Drosophila melanogaster, respectively. The Wb-HtrA has arthologues in several proteobacteria with very high homology and hence is highly conserved not only among Wolbachia of filarial parasites but also across proteobacteria. The phylogenetic tree constructed using Neighbor-Joining method showed two main clusters: cluster-I containing bacteria that dwell in diverse habitats such as soil, fresh and marine waters and plants and cluster-II comprising Anaplasma sp. and Erlichia, and Wolbachia endosymbionts of insects and nematodes, in distinct groups.

CONCLUSIONS: HtrA-type serine protease from Wolbachia of W. bancrofti is highly conserved among filarial parasites. It will be of interest to know whether filarial Wolbachia HtrA type of serine protease might influence apoptosis and lymphatic epithelium, thereby playing a role in the filarial pathogenesis. Such information will be useful for identifying targets for the development of newer drugs for filariasis treatment, especially for preventing lymphatic pathology.}, } @article {pmid21687646, year = {2011}, author = {Debrah, AY and Mand, S and Marfo-Debrekyei, Y and Batsa, L and Albers, A and Specht, S and Klarmann, U and Pfarr, K and Adjei, O and Hoerauf, A}, title = {Macrofilaricidal Activity in Wuchereria bancrofti after 2 Weeks Treatment with a Combination of Rifampicin plus Doxycycline.}, journal = {Journal of parasitology research}, volume = {2011}, number = {}, pages = {201617}, pmid = {21687646}, issn = {2090-0031}, abstract = {Infection with the filarial nematode Wuchereria bancrofti can lead to lymphedema, hydrocele, and elephantiasis. Since adult worms cause pathology in lymphatic filariasis (LF), it is imperative to discover macrofilaricidal drugs for the treatment of the infection. Endosymbiotic Wolbachia in filariae have emerged as a new target for antibiotics which can lead to macrofilaricidal effects. In Ghana, a pilot study was carried out with 39 LF-infected men; 12 were treated with 200 mg doxycycline/day for 4 weeks, 16 were treated with a combination of 200 mg doxycycline/day + 10 mg/kg/day rifampicin for 2 weeks, and 11 patients received placebo. Patients were monitored for Wolbachia and microfilaria loads, antigenaemia, and filarial dance sign (FDS). Both 4-week doxycycline and the 2-week combination treatment reduced Wolbachia load significantly. At 18 months posttreatment, four-week doxycycline resulted in 100% adult worm loss, and the 2-week combination treatment resulted in a 50% adult worm loss. In conclusion, this pilot study with a combination of 2-week doxycycline and rifampicin demonstrates moderate macrofilaricidal activity against W. bancrofti.}, } @article {pmid21676204, year = {2011}, author = {Petridis, M and Chatzidimitriou, D}, title = {Characterization of an intergenic polymorphic site (pp-hC1A_5) in Wolbachia pipientis (wPip).}, journal = {Molecular ecology resources}, volume = {11}, number = {4}, pages = {753-756}, doi = {10.1111/j.1755-0998.2011.02991.x}, pmid = {21676204}, issn = {1755-0998}, mesh = {Animals ; Cluster Analysis ; Culex/microbiology ; DNA Primers/genetics ; DNA, Bacterial/chemistry/genetics ; *DNA, Intergenic ; Genotype ; Molecular Sequence Data ; Phylogeography ; *Polymorphism, Genetic ; Sequence Analysis, DNA ; Wolbachia/*classification/*genetics/isolation & purification ; }, abstract = {Wolbachia pipientis (wPip) is an intracellular bacterium causing cytoplasmic incompatibility in arthropods, including mosquitoes of the Culex pipiens complex. Here, we present a method useful for genotyping within the wPip group. Primers were designed using a Tandem Repeat Finder program to amplify an intergenic, polymorphic site (pp-hC1A_5) of wPip. The polymorphic site is located between genes that code for polynucleotide phosphorylase and a hypothetical protein (C1A_5). Comparison of these wPip genomic regions from C. pipiens mosquitoes sampled in different geographic regions revealed deletions of fragments that proved useful in phylogenetic analysis.}, } @article {pmid21674166, year = {2011}, author = {Vashishtha, A and Sharama, KK and Lakhanpaul, S}, title = {Co-existence, phylogeny and putative role of Wolbachia and yeast-like symbiont (YLS) in Kerria lacca (Kerr).}, journal = {Current microbiology}, volume = {63}, number = {2}, pages = {206-212}, pmid = {21674166}, issn = {1432-0991}, mesh = {Animals ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Fungal/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Genes, rRNA ; Hemiptera/*microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Fungal/genetics ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 18S/genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; *Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/*physiology ; Yeasts/classification/genetics/*isolation & purification/*physiology ; }, abstract = {This study reports the first detection of Wolbachia and yeast-like symbiont (YLS) harbored in Kerria lacca (Kerr), a scale insect, latter of which produces an economically important natural resin, known as lac. Wolbachia was detected using PCR amplification and sequencing of 16S rDNA; and further confirmation and phylogenetic analysis was carried out by fast evolving wsp gene. Neighbor-joining and maximum parsimonious (MP) analysis showed that this strain belongs to subgroup "ori" of Wolbachia super group B of arthropods. Wolbachia of K. lacca is hereby designated as "wKerlac" according to Wolbachia nomenclature system. Histological study revealed the presence of yeast-like endosymbiont, which was also confirmed by PCR amplification of 18S rDNA. Phylogenetic analysis revealed that YLS of K. lacca is quite distinct from YLS of aphid, planthoppers, and beetles. Putative roles of Wolbachia in lecanoid chromosome system of sex determination and in biased sex ratio of K. lacca populations; and YLS in nutritional supplementation and detoxifying substances which are deleterious to K. lacca, are hereby, suggested.}, } @article {pmid21668807, year = {2011}, author = {Kraaijeveld, K and Franco, P and de Knijff, P and Stouthamer, R and van Alphen, JJ}, title = {Clonal genetic variation in a Wolbachia-infected asexual wasp: horizontal transmission or historical sex?.}, journal = {Molecular ecology}, volume = {20}, number = {17}, pages = {3644-3652}, doi = {10.1111/j.1365-294X.2011.05150.x}, pmid = {21668807}, issn = {1365-294X}, mesh = {Animals ; DNA, Mitochondrial/genetics ; Disease Transmission, Infectious ; Female ; *Genes, Bacterial ; Genetic Variation ; Haplotypes ; *Host-Pathogen Interactions ; Male ; Microsatellite Repeats ; Molecular Sequence Data ; Multilocus Sequence Typing/methods ; Parthenogenesis/genetics ; Phylogeography ; Sequence Analysis, DNA ; Sex Factors ; Wasps/*genetics/microbiology ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {Wolbachia are endocellular bacteria known for manipulating the reproductive systems of many of their invertebrate hosts. Wolbachia are transmitted vertically from mother to offspring. In addition, new infections result from horizontal transmission between different host species. However, to what extent horizontal transmission plays a role in the spread of a new infection through the host population is unknown. Here, we investigate whether horizontal transmission of Wolbachia can explain clonal genetic variation in natural populations of Leptopilina clavipes, a parasitoid wasp infected with a parthenogenesis-inducing Wolbachia. We assessed variance of markers on the nuclear, mitochondrial and Wolbachia genomes. The nuclear and mitochondrial markers displayed significant and congruent variation among thelytokous wasp lineages, showing that multiple lineages have become infected with Wolbachia. The alternative hypothesis in which a single female became infected, the daughters of which mated with males (thus introducing nuclear genetic variance) cannot account for the presence of concordant variance in mtDNA. All Wolbachia markers, including the hypervariable wsp gene, were invariant, suggesting that only a single strain of Wolbachia is involved. These results show that Wolbachia has transferred horizontally to infect multiple female lineages during the early spread through L. clavipes. Remarkably, multiple thelytokous lineages have persisted side by side in the field for tens of thousands of generations.}, } @article {pmid21663992, year = {2011}, author = {Cordaux, R and Bouchon, D and Grève, P}, title = {The impact of endosymbionts on the evolution of host sex-determination mechanisms.}, journal = {Trends in genetics : TIG}, volume = {27}, number = {8}, pages = {332-341}, doi = {10.1016/j.tig.2011.05.002}, pmid = {21663992}, issn = {0168-9525}, mesh = {Animals ; *Biological Evolution ; Diploidy ; Female ; Fertilization ; Haploidy ; Inheritance Patterns ; Isopoda/genetics/physiology ; Male ; Parthenogenesis ; Phenotype ; Sex Chromosomes/genetics ; *Sex Determination Processes ; Sex Ratio ; *Symbiosis ; Wolbachia/genetics/pathogenicity/physiology ; }, abstract = {The past years have revealed that inherited bacterial endosymbionts are important sources of evolutionary novelty for their eukaryotic hosts. In this review we discuss a fundamental biological process of eukaryotes influenced by bacterial endosymbionts: the mechanisms of sex determination. Because they are maternally inherited, several endosymbionts of arthropods, known as reproductive parasites, have developed strategies to convert non-transmitting male hosts into transmitting females through feminization of genetic males and parthenogenesis induction. Recent investigations have also highlighted that endosymbionts can impact upon host sex determination more subtly through genetic conflicts, resulting in selection of host nuclear genes resisting endosymbiont effects. Paradoxically, it is because of their selfish nature that reproductive parasites are such powerful agents of evolutionary change in their host sex-determination mechanisms. They might therefore represent excellent models for studying transitions between sex-determining systems and, more generally, the evolution of sex-determination mechanisms in eukaryotes.}, } @article {pmid21629728, year = {2011}, author = {Fischer, K and Beatty, WL and Jiang, D and Weil, GJ and Fischer, PU}, title = {Tissue and stage-specific distribution of Wolbachia in Brugia malayi.}, journal = {PLoS neglected tropical diseases}, volume = {5}, number = {5}, pages = {e1174}, pmid = {21629728}, issn = {1935-2735}, mesh = {Animal Structures/microbiology ; Animals ; Antibodies, Bacterial/immunology/isolation & purification ; Antibodies, Monoclonal/immunology/isolation & purification ; Brugia malayi/*growth & development/*microbiology/physiology ; Female ; Humans ; Immunohistochemistry/methods ; In Situ Hybridization, Fluorescence/methods ; Male ; Microscopy/methods ; Parasitology/methods ; *Symbiosis ; Wolbachia/*growth & development/*isolation & purification/physiology ; }, abstract = {BACKGROUND: Most filarial parasite species contain Wolbachia, obligatory bacterial endosymbionts that are crucial for filarial development and reproduction. They are targets for alternative chemotherapy, but their role in the biology of filarial nematodes is not well understood. Light microscopy provides important information on morphology, localization and potential function of these bacteria. Surprisingly, immunohistology and in situ hybridization techniques have not been widely used to monitor Wolbachia distribution during the filarial life cycle.

METHODS/PRINCIPAL FINDINGS: A monoclonal antibody directed against Wolbachia surface protein and in situ hybridization targeting Wolbachia 16S rRNA were used to monitor Wolbachia during the life cycle of B. malayi. In microfilariae and vector stage larvae only a few cells contain Wolbachia. In contrast, large numbers of Wolbachia were detected in the lateral chords of L4 larvae, but no endobacteria were detected in the genital primordium. In young adult worms (5 weeks p.i.), a massive expansion of Wolbachia was observed in the lateral chords adjacent to ovaries or testis, but no endobacteria were detected in the growth zone of the ovaries, uterus, the growth zone of the testis or the vas deferens. Confocal laser scanning and transmission electron microscopy showed that numerous Wolbachia are aligned towards the developing ovaries and single endobacteria were detected in the germline. In inseminated females (8 weeks p.i.) Wolbachia were observed in the ovaries, embryos and in decreasing numbers in the lateral chords. In young males Wolbachia were found in distinct zones of the testis and in large numbers in the lateral chords in the vicinity of testicular tissue but never in mature spermatids or spermatozoa.

CONCLUSIONS: Immunohistology and in situ hybridization show distinct tissue and stage specific distribution patterns for Wolbachia in B. malayi. Extensive multiplication of Wolbachia occurs in the lateral chords of L4 and young adults adjacent to germline cells.}, } @article {pmid21625582, year = {2011}, author = {Hughes, GL and Koga, R and Xue, P and Fukatsu, T and Rasgon, JL}, title = {Wolbachia infections are virulent and inhibit the human malaria parasite Plasmodium falciparum in Anopheles gambiae.}, journal = {PLoS pathogens}, volume = {7}, number = {5}, pages = {e1002043}, pmid = {21625582}, issn = {1553-7374}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/immunology/*microbiology/*parasitology ; Gene Expression ; Host-Parasite Interactions ; Humans ; In Situ Hybridization, Fluorescence ; Malaria, Falciparum/parasitology/prevention & control ; Oocysts/growth & development ; Plasmodium falciparum/*growth & development/immunology ; Polymerase Chain Reaction ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Endosymbiotic Wolbachia bacteria are potent modulators of pathogen infection and transmission in multiple naturally and artificially infected insect species, including important vectors of human pathogens. Anopheles mosquitoes are naturally uninfected with Wolbachia, and stable artificial infections have not yet succeeded in this genus. Recent techniques have enabled establishment of somatic Wolbachia infections in Anopheles. Here, we characterize somatic infections of two diverse Wolbachia strains (wMelPop and wAlbB) in Anopheles gambiae, the major vector of human malaria. After infection, wMelPop disseminates widely in the mosquito, infecting the fat body, head, sensory organs and other tissues but is notably absent from the midgut and ovaries. Wolbachia initially induces the mosquito immune system, coincident with initial clearing of the infection, but then suppresses expression of immune genes, coincident with Wolbachia replication in the mosquito. Both wMelPop and wAlbB significantly inhibit Plasmodium falciparum oocyst levels in the mosquito midgut. Although not virulent in non-bloodfed mosquitoes, wMelPop exhibits a novel phenotype and is extremely virulent for approximately 12-24 hours post-bloodmeal, after which surviving mosquitoes exhibit similar mortality trajectories to control mosquitoes. The data suggest that if stable transinfections act in a similar manner to somatic infections, Wolbachia could potentially be used as part of a strategy to control the Anopheles mosquitoes that transmit malaria.}, } @article {pmid21622788, year = {2011}, author = {Casper-Lindley, C and Kimura, S and Saxton, DS and Essaw, Y and Simpson, I and Tan, V and Sullivan, W}, title = {Rapid fluorescence-based screening for Wolbachia endosymbionts in Drosophila germ line and somatic tissues.}, journal = {Applied and environmental microbiology}, volume = {77}, number = {14}, pages = {4788-4794}, pmid = {21622788}, issn = {1098-5336}, mesh = {Animals ; DNA, Bacterial/analysis ; Drosophila melanogaster/*microbiology ; Germ Cells/microbiology ; Microscopy, Fluorescence/*methods ; Nucleic Acids/metabolism ; Polymerase Chain Reaction ; Spiroplasma/genetics/isolation & purification ; *Symbiosis ; Time Factors ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia is a globally distributed bacterial endosymbiont present in arthropods and nematodes. The advent of sensitive PCR-based approaches has greatly facilitated the identification of Wolbachia-infected individuals and analysis of population infection levels. Here, a complementary visual fluorescence-based Wolbachia screening approach is described. Through the use of the fluorescent dye Syto-11, Wolbachia can be efficiently detected in various Drosophila tissues, including ovaries. Syto-11 also stains Wolbachia in other insects. Because Wolbachia is inherited through the maternal germ line, bacteria reside in the ovaries of flies in infected populations. An advantage of this staining approach is that it informs about Wolbachia titer as well as its tissue and cellular distribution. Using this method, the infection status of insect populations in two central California locations was determined, and variants with unusually low or high Wolbachia titers were isolated. In addition, a variant with ovarioles containing both infected and uninfected egg chambers was identified. Syto-11 staining of Cardinium- and Spiroplasma-infected insects was also analyzed.}, } @article {pmid21612535, year = {2011}, author = {Foongladda, S and Inthawong, D and Kositanont, U and Gaywee, J}, title = {Rickettsia, Ehrlichia, Anaplasma, and Bartonella in ticks and fleas from dogs and cats in Bangkok.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {11}, number = {10}, pages = {1335-1341}, doi = {10.1089/vbz.2010.0174}, pmid = {21612535}, issn = {1557-7759}, mesh = {Anaplasma/genetics/*isolation & purification ; Animals ; Bartonella/genetics/*isolation & purification ; Cats ; Ctenocephalides/*microbiology ; DNA Primers ; DNA, Bacterial/chemistry/genetics ; Dogs ; Ehrlichia/genetics/*isolation & purification ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Rhipicephalus/*microbiology ; Rickettsia/genetics/*isolation & purification ; Sequence Analysis, DNA ; Species Specificity ; Thailand ; }, abstract = {Flea and tick specimens (5-10 fleas or ticks) on dogs and cats from various sites in Bangkok were tested by polymerase chain reaction and DNA sequencing to detect DNA of bacteria Rickettsia (gltA and 17 kDa genes), Anaplasmataceae (16S rRNA gene), and Bartonella (pap31 and its genes). We confirmed that Rickettsia sp. related to Rickettsia felis was detected in 66 of 98 (67.4%) flea specimens from dogs, whereas 8 Bartonella henselae and 2 Bartonella clarridgeiae were detected in 10 of 54 (18.5%) flea specimens from cats. Further, this work provides the first evidence of 10 Ehrlichia canis (3.3%), 7 Anaplasma platys (2.3%), and 2 Wolbachia spp. (0.66%) in 304 Rhipicephalus sanguineus tick specimens in Thailand.}, } @article {pmid21612526, year = {2011}, author = {Wong, SS and Teng, JL and Poon, RW and Choi, GK and Chan, KH and Yeung, ML and Hui, JJ and Yuen, KY}, title = {Comparative evaluation of a point-of-care immunochromatographic test SNAP 4Dx with molecular detection tests for vector-borne canine pathogens in Hong Kong.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {11}, number = {9}, pages = {1269-1277}, doi = {10.1089/vbz.2010.0265}, pmid = {21612526}, issn = {1557-7759}, mesh = {Animals ; Arthropod Vectors/microbiology ; Babesia/immunology ; Chromatography, Affinity/*standards ; DNA Primers ; Databases, Nucleic Acid ; Dirofilaria immitis/immunology ; Dog Diseases/blood/*diagnosis/epidemiology/*parasitology ; Dogs ; Ehrlichia/immunology ; Hong Kong/epidemiology ; Point-of-Care Systems ; Polymerase Chain Reaction/*standards ; Reproducibility of Results ; }, abstract = {There are no comprehensive studies on the performance of commonly used point-of-care diagnostic enzyme immunoassay for common arthropod-borne canine pathogens. A comparative evaluation of an immunochromatographic test for these infections with a comprehensive polymerase chain reaction (PCR) test panel was performed on 100 pet dogs and 100 stray dogs without obvious clinical symptoms. Of the 162 positive test results from both immunochromatographic test and PCR, there was 85.2% concordance. The 24 discordant results between serology and PCR occurred in tests involving Ehrlichia canis (14) and Anaplasma platys (10), which may be related to the time of infection. No positive cases of borreliosis or rickettsiosis were detected. One important limitation of the immunochromatographic test was its lack of testing for babesiosis and hepatozoonosis. The former is the most prevalent arthropod-borne canine infection in our cohort (41%). Coinfections were found in 19% stray dogs and 6% of pet dogs with both tests (p < 0.01). Seventeen and 8 samples from stray and pet dogs, respectively, were initially positive in the PCR test for Ehrlichia. However, on sequencing of the PCR amplicon, 10 from stray and 2 from pet dogs were found to be Wolbachia sequences instead, with 100% nucleotide identity to the 16S rRNA sequence of Wolbachia endosymbiont of Dirofilaria immitis. The presence of Wolbachia DNAemia (6%) correlated well with the molecular test and immunochromatographic antigen test for D. immitis.}, } @article {pmid21606368, year = {2011}, author = {Bennuru, S and Meng, Z and Ribeiro, JM and Semnani, RT and Ghedin, E and Chan, K and Lucas, DA and Veenstra, TD and Nutman, TB}, title = {Stage-specific proteomic expression patterns of the human filarial parasite Brugia malayi and its endosymbiont Wolbachia.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {108}, number = {23}, pages = {9649-9654}, pmid = {21606368}, issn = {1091-6490}, support = {HHSN261200800001C/RC/CCR NIH HHS/United States ; HHSN261200800001E/CA/NCI NIH HHS/United States ; /ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/*analysis/classification ; Brugia malayi/growth & development/*metabolism/microbiology ; Chromatography, Liquid/methods ; Cluster Analysis ; Female ; Filariasis/parasitology ; Helminth Proteins/*analysis/classification ; Host-Pathogen Interactions ; Humans ; Larva/growth & development/metabolism/microbiology ; Life Cycle Stages ; Male ; Proteome/*analysis/classification ; Proteomics/*methods ; Symbiosis ; Tandem Mass Spectrometry ; Wolbachia/*metabolism/physiology ; }, abstract = {Global proteomic analyses of pathogens have thus far been limited to unicellular organisms (e.g., protozoa and bacteria). Proteomic analyses of most eukaryotic pathogens (e.g., helminths) have been restricted to specific organs, specific stages, or secretomes. We report here a large-scale proteomic characterization of almost all the major mammalian stages of Brugia malayi, a causative agent of lymphatic filariasis, resulting in the identification of more than 62% of the products predicted from the Bm draft genome. The analysis also yielded much of the proteome of Wolbachia, the obligate endosymbiont of Bm that also expressed proteins in a stage-specific manner. Of the 11,610 predicted Bm gene products, 7,103 were definitively identified from adult male, adult female, blood-borne and uterine microfilariae, and infective L3 larvae. Among the 4,956 gene products (42.5%) inferred from the genome as "hypothetical," the present study was able to confirm 2,336 (47.1%) as bona fide proteins. Analysis of protein families and domains coupled with stage-specific expression highlight the important pathways that benefit the parasite during its development in the host. Gene set enrichment analysis identified extracellular matrix proteins and those with immunologic effects as enriched in the microfilarial and L3 stages. Parasite sex- and stage-specific protein expression identified those pathways related to parasite differentiation and demonstrates stage-specific expression by the Bm endosymbiont Wolbachia as well.}, } @article {pmid21598659, year = {2011}, author = {Belousov, AO and Kozeretskaia, IA}, title = {[Symbiotic bacteria, which modify reproduction processes of Drosophila melanogaster].}, journal = {Mikrobiolohichnyi zhurnal (Kiev, Ukraine : 1993)}, volume = {73}, number = {2}, pages = {43-52}, pmid = {21598659}, issn = {1028-0987}, mesh = {Animals ; Bacteroidetes/*growth & development ; Drosophila melanogaster/growth & development/*microbiology/physiology ; Female ; Male ; Reproduction ; Spiroplasma/*growth & development ; Symbiosis ; Wolbachia/*growth & development ; }, abstract = {Cytoplasmic bacteria-symbionts are actively investigated all over the world for the last ten years. The scale of their spreading in natural and laboratory populations of invertebrates, especially arthropods, is impressing scientists' imagination. Ways of their intraspecific and interspecific transmission are various. The nature and mechanisms of their interaction both between themselves and with their hosts are extremely diverse. Cytoplasmic incompatibility, parthenogenesis, male-killing, feminization, tissue degeneration and others are those phenotypic effects, which intracellular bacteria can cause in their hosts. Moreover, the same bacterium can have diverse influences on different hosts. So it is possible to talk about many-sidedness of endosymbiont interaction with a host, which is determined both by bacterium and host biology. However, majority of phenotypic effects is directed to the successful colonization of hosts' populations by increasing the number of infected females. Alas, the mechanisms of interaction are still not enough investigated. Besides, intracellular bacteria-symbionts can be one of the factors of the so-called "infectious species formation". In any investigation, model organisms, Drosophila melanogaster belonging to them, take a special place. Among drosophila's endosymbionts, only bacteria from genera Wolbachia, Spiroplasma and Cardinium were broadly investigated: their spreading in Drosophila melanogaster populations all over the world, the infecting level and those phenotypic effects, which they can cause. So this review is dedicated to analysis of these studies.}, } @article {pmid21595768, year = {2011}, author = {Dyer, KA and Burke, C and Jaenike, J}, title = {Wolbachia-mediated persistence of mtDNA from a potentially extinct species.}, journal = {Molecular ecology}, volume = {20}, number = {13}, pages = {2805-2817}, doi = {10.1111/j.1365-294X.2011.05128.x}, pmid = {21595768}, issn = {1365-294X}, mesh = {Animals ; Base Sequence ; Biological Evolution ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics/microbiology ; Female ; Gene Transfer, Horizontal ; Haplotypes ; Maine ; Models, Genetic ; Molecular Sequence Data ; Multilocus Sequence Typing ; New York ; Pennsylvania ; Phylogeny ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Symbiosis ; Time Factors ; Wolbachia/*genetics ; }, abstract = {Drosophila quinaria is polymorphic for infection with Wolbachia, a maternally transmitted endosymbiont. Wolbachia-infected individuals carry mtDNA that is only distantly related to the mtDNA of uninfected individuals, and the clade encompassing all mtDNA haplotypes within D. quinaria also includes the mtDNA of several other species of Drosophila. Nuclear gene variation reveals no difference between the Wolbachia-infected and uninfected individuals of D. quinaria, indicating that they all belong to the same interbreeding biological species. We suggest that the Wolbachia and the mtDNA with which it is associated were derived via interspecific hybridization and introgression. The sequences in the Wolbachia and the associated mtDNA are ≥6% divergent from those of any known Drosophila species. Thus, in spite of nearly complete species sampling, the sequences from which these mitochondria were derived remain unknown, raising the possibility that the donor species is extinct. The association between Wolbachia infection and mtDNA type within D. quinaria suggests that Wolbachia may be required for the continued persistence of the mtDNA from an otherwise extinct Drosophila species. We hypothesize that pathogen-protective effects conferred by Wolbachia operate in a negative frequency-dependent manner, thus bringing about a stable polymorphism for Wolbachia infection.}, } @article {pmid21584395, year = {2011}, author = {Yun, Y and Peng, Y and Liu, FX and Lei, C}, title = {Wolbachia screening in spiders and assessment of horizontal transmission between predator and prey.}, journal = {Neotropical entomology}, volume = {40}, number = {2}, pages = {164-169}, pmid = {21584395}, issn = {1678-8052}, mesh = {Animals ; Bacterial Infections/transmission/veterinary ; Spiders/*microbiology ; Wolbachia/*isolation & purification ; }, abstract = {Recent studies have revealed that the prevalence of Wolbachia in arthropods is attributable not only to its vertical transmission, but also to its horizontal transfer. In order to assess the horizontal transmission of Wolbachia between predator and prey, arthropods belonging to 11 spider families and six insect families were collected in the same field of rice. The distribution of Wolbachia in these arthropods was detected by diagnostic PCR amplification of the wsp (Wolbachia outer surface protein gene) and 16S rDNA genes. Nurscia albofasciata Strand (Araneae: Titanoecidae), Propylea japonica Thunberg (Coleoptera: Coccinellidae), Paederus fuscipes Curtis (Coleoptera: Staphylinidae), and Nilaparvata lugens Stal (Homoptera: Delphacidae) were infected with Wolbachia. This is the first report of infection of N. albofasciata and P. fuscipes by Wolbachia. No direct evidence indicated the existence of horizontal transmission of Wolbachia between predator and prey.}, } @article {pmid21576469, year = {2011}, author = {Hussain, M and Frentiu, FD and Moreira, LA and O'Neill, SL and Asgari, S}, title = {Wolbachia uses host microRNAs to manipulate host gene expression and facilitate colonization of the dengue vector Aedes aegypti.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {108}, number = {22}, pages = {9250-9255}, pmid = {21576469}, issn = {1091-6490}, mesh = {Aedes/*metabolism ; Animals ; Dengue/*transmission ; *Gene Expression Regulation ; Gene Silencing ; Green Fluorescent Proteins/metabolism ; Insect Vectors/metabolism ; MicroRNAs/*genetics/metabolism ; Models, Biological ; RNA Interference ; Time Factors ; Virus Replication ; Wolbachia/*metabolism ; }, abstract = {The obligate endosymbiont Wolbachia pipientis is found in a wide range of invertebrates where they are best known for manipulating host reproduction. Recent studies have shown that Wolbachia also can modulate the lifespan of host insects and interfere with the development of human pathogens in mosquito vectors. Despite considerable study, very little is known about the molecular interactions between Wolbachia and its hosts that might mediate these effects. Using microarrays, we show that the microRNA (miRNA) profile of the mosquito, Aedes aegypti, is significantly altered by the wMelPop-CLA strain of W. pipientis. We found that a host miRNA (aae-miR-2940) is induced after Wolbachia infection in both mosquitoes and cell lines. One target of aae-miR-2940 is the Ae. aegypti metalloprotease gene. Interestingly, expression of the target gene was induced after Wolbachia infection, ectopic expression of the miRNA independent of Wolbachia, or transfection of an artificial mimic of the miRNA into mosquito cells. We also confirmed the interaction of aae-miR-2940 with the target sequences using GFP as a reporter gene. Silencing of the metalloprotease gene in both Wolbachia-infected cells and adult mosquitoes led to a significant reduction in Wolbachia density, as did inhibition of the miRNA in cells. These results indicate that manipulation of the mosquito metalloprotease gene via aae-miR-2940 is crucial for efficient maintenance of the endosymbiont. This report shows how Wolbachia alters the host miRNA profile and provides insight into the mechanisms of host manipulation used by this widespread endosymbiont.}, } @article {pmid21573076, year = {2011}, author = {Saridaki, A and Sapountzis, P and Harris, HL and Batista, PD and Biliske, JA and Pavlikaki, H and Oehler, S and Savakis, C and Braig, HR and Bourtzis, K}, title = {Wolbachia prophage DNA adenine methyltransferase genes in different Drosophila-Wolbachia associations.}, journal = {PloS one}, volume = {6}, number = {5}, pages = {e19708}, pmid = {21573076}, issn = {1932-6203}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Amino Acid Sequence ; Animals ; Base Sequence ; Blotting, Southern ; Drosophila/*microbiology ; Genes, Viral/*genetics ; Genome, Bacterial/genetics ; Host-Pathogen Interactions/*genetics ; Molecular Sequence Data ; Open Reading Frames/genetics ; Phylogeny ; Polymerase Chain Reaction ; Prophages/*enzymology/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; Site-Specific DNA-Methyltransferase (Adenine-Specific)/chemistry/*genetics ; Wolbachia/genetics/*virology ; }, abstract = {Wolbachia is an obligatory intracellular bacterium which often manipulates the reproduction of its insect and isopod hosts. In contrast, Wolbachia is an essential symbiont in filarial nematodes. Lately, Wolbachia has been implicated in genomic imprinting of host DNA through cytosine methylation. The importance of DNA methylation in cell fate and biology calls for in depth studying of putative methylation-related genes. We present a molecular and phylogenetic analysis of a putative DNA adenine methyltransferase encoded by a prophage in the Wolbachia genome. Two slightly different copies of the gene, met1 and met2, exhibit a different distribution over various Wolbachia strains. The met2 gene is present in the majority of strains, in wAu, however, it contains a frameshift caused by a 2 bp deletion. Phylogenetic analysis of the met2 DNA sequences suggests a long association of the gene with the Wolbachia host strains. In addition, our analysis provides evidence for previously unnoticed multiple infections, the detection of which is critical for the molecular elucidation of modification and/or rescue mechanism of cytoplasmic incompatibility.}, } @article {pmid21572955, year = {2011}, author = {Bossan, B and Koehncke, A and Hammerstein, P}, title = {A new model and method for understanding Wolbachia-induced cytoplasmic incompatibility.}, journal = {PloS one}, volume = {6}, number = {5}, pages = {e19757}, pmid = {21572955}, issn = {1932-6203}, mesh = {Animals ; Biological Evolution ; Cytoplasm/*microbiology ; Female ; Host-Parasite Interactions/*physiology ; Insecta/*microbiology ; Male ; *Models, Biological ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular bacteria transmitted almost exclusively vertically through eggs. In response to this mode of transmission, Wolbachia strategically manipulate their insect hosts' reproduction. In the most common manipulation type, cytoplasmic incompatibility, infected males can only mate with infected females, but infected females can mate with all males. The mechanism of cytoplasmic incompatibility is unknown; theoretical and empirical findings need to converge to broaden our understanding of this phenomenon. For this purpose, two prominent models have been proposed: the mistiming-model and the lock-key-model. The former states that Wolbachia manipulate sperm of infected males to induce a fatal delay of the male pronucleus during the first embryonic division, but that the bacteria can compensate the delay by slowing down mitosis in fertilized eggs. The latter states that Wolbachia deposit damaging "locks" on sperm DNA of infected males, but can also provide matching "keys" in infected eggs to undo the damage. The lock-key-model, however, needs to assume a large number of locks and keys to explain all existing incompatibility patterns. The mistiming-model requires fewer assumptions but has been contradicted by empirical results. We therefore expand the mistiming-model by one quantitative dimension to create the new, so-called goalkeeper-model. Using a method based on formal logic, we show that both lock-key- and goalkeeper-model are consistent with existing data. Compared to the lock-key-model, however, the goalkeeper-model assumes only two factors and provides an idea of the evolutionary emergence of cytoplasmic incompatibility. Available cytological evidence suggests that the hypothesized second factor of the goalkeeper-model may indeed exist. Finally, we suggest empirical tests that would allow to distinguish between the models. Generalizing our results might prove interesting for the study of the mechanism and evolution of other host-parasite interactions.}, } @article {pmid21559467, year = {2011}, author = {Narzisi, G and Mishra, B}, title = {Comparing de novo genome assembly: the long and short of it.}, journal = {PloS one}, volume = {6}, number = {4}, pages = {e19175}, pmid = {21559467}, issn = {1932-6203}, mesh = {Algorithms ; Biotechnology/methods ; Computational Biology/methods ; Genetic Techniques ; *Genome ; Genome, Bacterial ; Genome, Human ; Genome-Wide Association Study/methods ; Genomics ; Humans ; ROC Curve ; Reproducibility of Results ; Sequence Analysis, DNA/*methods ; Software ; }, abstract = {Recent advances in DNA sequencing technology and their focal role in Genome Wide Association Studies (GWAS) have rekindled a growing interest in the whole-genome sequence assembly (WGSA) problem, thereby, inundating the field with a plethora of new formalizations, algorithms, heuristics and implementations. And yet, scant attention has been paid to comparative assessments of these assemblers' quality and accuracy. No commonly accepted and standardized method for comparison exists yet. Even worse, widely used metrics to compare the assembled sequences emphasize only size, poorly capturing the contig quality and accuracy. This paper addresses these concerns: it highlights common anomalies in assembly accuracy through a rigorous study of several assemblers, compared under both standard metrics (N50, coverage, contig sizes, etc.) as well as a more comprehensive metric (Feature-Response Curves, FRC) that is introduced here; FRC transparently captures the trade-offs between contigs' quality against their sizes. For this purpose, most of the publicly available major sequence assemblers--both for low-coverage long (Sanger) and high-coverage short (Illumina) reads technologies--are compared. These assemblers are applied to microbial (Escherichia coli, Brucella, Wolbachia, Staphylococcus, Helicobacter) and partial human genome sequences (Chr. Y), using sequence reads of various read-lengths, coverages, accuracies, and with and without mate-pairs. It is hoped that, based on these evaluations, computational biologists will identify innovative sequence assembly paradigms, bioinformaticists will determine promising approaches for developing "next-generation" assemblers, and biotechnologists will formulate more meaningful design desiderata for sequencing technology platforms. A new software tool for computing the FRC metric has been developed and is available through the AMOS open-source consortium.}, } @article {pmid21559343, year = {2011}, author = {Zheng, Y and Ren, PP and Wang, JL and Wang, YF}, title = {Wolbachia-induced cytoplasmic incompatibility is associated with decreased Hira expression in male Drosophila.}, journal = {PloS one}, volume = {6}, number = {4}, pages = {e19512}, pmid = {21559343}, issn = {1932-6203}, mesh = {Animals ; Cell Cycle Proteins/biosynthesis/*physiology ; Crosses, Genetic ; Cytoplasm/*metabolism/microbiology ; Drosophila Proteins/biosynthesis/*physiology ; Drosophila melanogaster ; Female ; Fertility/genetics ; Histone Chaperones/biosynthesis/*physiology ; Male ; Microscopy, Fluorescence/methods ; Mutation ; Phenotype ; Reproduction ; Reverse Transcriptase Polymerase Chain Reaction ; Rickettsiaceae Infections/*metabolism ; Time Factors ; Transcription Factors/biosynthesis/*physiology ; Wolbachia/*metabolism ; }, abstract = {BACKGROUND: Wolbachia are obligate endosymbiotic bacteria that infect numerous species of arthropods and nematodes. Wolbachia can induce several reproductive phenotypes in their insect hosts including feminization, male-killing, parthenogenesis and cytoplasmic incompatibility (CI). CI is the most common phenotype and occurs when Wolbachia-infected males mate with uninfected females resulting in no or very low numbers of viable offspring. However, matings between males and females infected with the same strain of Wolbachia result in viable progeny. Despite substantial scientific effort, the molecular mechanisms underlying CI are currently unknown.

Gene expression studies were undertaken in Drosophila melanogaster and D. simulans which display differential levels of CI using quantitative RT-PCR. We show that Hira expression is correlated with the induction of CI and occurs in a sex-specific manner. Hira expression is significantly lower in males which induce strong CI when compared to males inducing no CI or Wolbachia-uninfected males. A reduction in Hira expression is also observed in 1-day-old males that induce stronger CI compared to 5-day-old males that induce weak or no CI. In addition, Hira mutated D. melanogaster males mated to uninfected females result in significantly decreased hatch rates comparing with uninfected crosses. Interestingly, wMel-infected females may rescue the hatch rates. An obvious CI phenotype with chromatin bridges are observed in the early embryo resulting from Hira mutant fertilization, which strongly mimics the defects associated with CI.

CONCLUSIONS/SIGNIFICANCE: Our results suggest Wolbachia-induced CI in Drosophila occurs due to a reduction in Hira expression in Wolbachia-infected males leading to detrimental effects on sperm fertility resulting in embryo lethality. These results may help determine the underlying mechanism of CI and provide further insight in to the important role Hira plays in the interaction of Wolbachia and its insect host.}, } @article {pmid21546911, year = {2011}, author = {Iturbe-Ormaetxe, I and Walker, T and O' Neill, SL}, title = {Wolbachia and the biological control of mosquito-borne disease.}, journal = {EMBO reports}, volume = {12}, number = {6}, pages = {508-518}, pmid = {21546911}, issn = {1469-3178}, mesh = {Animals ; Australia ; Culicidae/*microbiology ; Dengue/prevention & control/transmission ; Genetic Fitness ; Insect Vectors/*microbiology ; Malaria/prevention & control/transmission ; *Mosquito Control ; Wolbachia/genetics/*physiology ; }, abstract = {Mosquito-borne diseases such as malaria, dengue fever and filariasis cause an enormous health burden to people living in tropical and subtropical regions of the world. Despite years of intense effort to control them, many of these diseases are increasing in prevalence, geographical distribution and severity, and options to control them are limited. The transinfection of mosquitos with the maternally inherited, endosymbiotic bacteria Wolbachia is a promising new biocontrol approach. Fruit fly Wolbachia strains can invade and sustain themselves in mosquito populations, reduce adult lifespan, affect mosquito reproduction and interfere with pathogen replication. Wolbachia-infected Aedes aegypti mosquitoes have been released in areas of Australia in which outbreaks of dengue fever occur, as a prelude to the application of this technology in dengue-endemic areas of south-east Asia.}, } @article {pmid21541357, year = {2011}, author = {Hancock, PA and Sinkins, SP and Godfray, HC}, title = {Strategies for introducing Wolbachia to reduce transmission of mosquito-borne diseases.}, journal = {PLoS neglected tropical diseases}, volume = {5}, number = {4}, pages = {e1024}, pmid = {21541357}, issn = {1935-2735}, support = {079059/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Culicidae/*microbiology ; Female ; Insect Control/*methods ; Male ; Models, Statistical ; Pest Control, Biological/*methods ; Wolbachia/*pathogenicity ; }, abstract = {Certain strains of the endosymbiont Wolbachia have the potential to lower the vectorial capacity of mosquito populations and assist in controlling a number of mosquito-borne diseases. An important consideration when introducing Wolbachia-carrying mosquitoes into natural populations is the minimisation of any transient increase in disease risk or biting nuisance. This may be achieved by predominantly releasing male mosquitoes. To explore this, we use a sex-structured model of Wolbachia-mosquito interactions. We first show that Wolbachia spread can be initiated with very few infected females provided the infection frequency in males exceeds a threshold. We then consider realistic introduction scenarios involving the release of batches of infected mosquitoes, incorporating seasonal fluctuations in population size. For a range of assumptions about mosquito population dynamics we find that male-biased releases allow the infection to spread after the introduction of low numbers of females, many fewer than with equal sex-ratio releases. We extend the model to estimate the transmission rate of a mosquito-borne pathogen over the course of Wolbachia establishment. For a range of release strategies we demonstrate that male-biased release of Wolbachia-infected mosquitoes can cause substantial transmission reductions without transiently increasing disease risk. The results show the importance of including mosquito population dynamics in studying Wolbachia spread and that male-biased releases can be an effective and safe way of rapidly establishing the symbiont in mosquito populations.}, } @article {pmid21533137, year = {2011}, author = {Chevalier, F and Herbinière-Gaboreau, J and Bertaux, J and Raimond, M and Morel, F and Bouchon, D and Grève, P and Braquart-Varnier, C}, title = {The immune cellular effectors of terrestrial isopod Armadillidium vulgare: meeting with their invaders, Wolbachia.}, journal = {PloS one}, volume = {6}, number = {4}, pages = {e18531}, pmid = {21533137}, issn = {1932-6203}, mesh = {Animals ; Flow Cytometry ; Hemocytes/ultrastructure ; In Situ Hybridization, Fluorescence ; Isopoda/*immunology/microbiology ; Microscopy, Electron, Transmission ; Phagocytosis ; Wolbachia/*immunology ; }, abstract = {BACKGROUND: Most of crustacean immune responses are well described for the aquatic forms whereas almost nothing is known for the isopods that evolved a terrestrial lifestyle. The latter are also infected at a high prevalence with Wolbachia, an endosymbiotic bacterium which affects the host immune system, possibly to improve its transmission. In contrast with insect models, the isopod Armadillidium vulgare is known to harbor Wolbachia inside the haemocytes.

In A. vulgare we characterized three haemocyte types (TEM, flow cytometry): the hyaline and semi-granular haemocytes were phagocytes, while semi-granular and granular haemocytes performed encapsulation. They were produced in the haematopoietic organs, from central stem cells, maturing as they moved toward the edge (TEM). In infected individuals, live Wolbachia (FISH) colonized 38% of the haemocytes but with low, variable densities (6.45±0.46 Wolbachia on average). So far they were not found in hyaline haemocytes (TEM). The haematopoietic organs contained 7.6±0.7×10(3)Wolbachia, both in stem cells and differentiating cells (FISH). While infected and uninfected one-year-old individuals had the same haemocyte density, in infected animals the proportion of granular haemocytes in particular decreased by one third (flow cytometry, Pearson's test = 12 822.98, df = 2, p<0.001).

CONCLUSIONS/SIGNIFICANCE: The characteristics of the isopod immune system fell within the range of those known from aquatic crustaceans. The colonization of the haemocytes by Wolbachia seemed to stand from the haematopoietic organs, which may act as a reservoir to discharge Wolbachia in the haemolymph, a known route for horizontal transfer. Wolbachia infection did not affect the haemocyte density, but the quantity of granular haemocytes decreased by one third. This may account for the reduced prophenoloxidase activity observed previously in these animals.}, } @article {pmid21526927, year = {2011}, author = {Bouwma, AM and Shoemaker, D}, title = {Wolbachia wSinvictaA infections in natural populations of the fire ant Solenopsis invicta: testing for phenotypic effects.}, journal = {Journal of insect science (Online)}, volume = {11}, number = {}, pages = {11}, pmid = {21526927}, issn = {1536-2442}, mesh = {Animals ; Ants/*microbiology/*physiology ; Argentina ; Base Sequence ; Female ; *Host-Pathogen Interactions ; Male ; Membrane Proteins/genetics ; Molecular Sequence Data ; *Phenotype ; Polymerase Chain Reaction ; Population Dynamics ; Sequence Analysis, DNA ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are intracellular bacteria that commonly infect many arthropods and some nematodes. In arthropods, these maternally transmitted bacteria often induce a variety of phenotypic effects to enhance their own spread within host populations. Wolbachia phenotypic effects generally either provide benefits to infected host females (cytoplasmic incompatibility, positive fitness effects) or bias host sex ratio in favor of females (male-killing, parthenogenesis, feminization), all of which increase the relative production of infected females in host populations. Wolbachia surveys have found infections to be exceedingly common in ants, but little is known at this juncture as to what phenotypic effects, if any, they induce in this group. Previous studies have demonstrated that individuals from native populations of the invasive fire ant Solenopsis invicta commonly harbor one or more of three Wolbachia variants. One of the variants, wSinvictaA, typically occurs at low prevalence in S. invicta populations, appears to have been transmitted horizontally into S. invicta three or more times, and has been lost repeatedly from host lineages over time. In order to determine the phenotypic effects and likely population dynamics of wSinvictaA infections in these ants, brood production patterns of newly mated fire ant queens were studied during simulated claustral founding and measured wSinvictaA transmission fidelity within mature single-queen families. No clear evidence was found for Wolbachia-induced cytoplasmic incompatibility, significant fitness effects, or male-killing. Maternal transmission was perfect to both virgin queens and males. Possible mechanisms for how this variant could be maintained in host populations are discussed.}, } @article {pmid21525789, year = {2011}, author = {Ringo, J and Sharon, G and Segal, D}, title = {Bacteria-induced sexual isolation in Drosophila.}, journal = {Fly}, volume = {5}, number = {4}, pages = {310-315}, doi = {10.4161/fly.5.4.15835}, pmid = {21525789}, issn = {1933-6942}, mesh = {Animals ; Cues ; Drosophila melanogaster/genetics/*microbiology/physiology ; Genetic Speciation ; *Mating Preference, Animal ; Reproduction ; *Social Isolation ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Commensal bacteria can induce sexual isolation between populations of Drosophila. This phenomenon has implications for speciation, and raises questions about its behavioral and developmental mechanisms, which are not yet known. In this Extra View, we discuss related work by others, bearing directly on these issues, and we speculate about how bacteria might influence fly behavior. There are many reports of interaction between Drosophila and their microbiota that significantly impacts mating preferences. Sexual isolation can be enhanced or reduced by altering the culture media, or the microbiota inhabiting those media. More dramatically, the endoparasite Wolbachia has induced strong mate preferences in some instances. While a sudden, ecologically induced shift in mating preferences falls far short of the changes required for speciation, it might be a first step in that direction. We hypothesize that bacteria-induced sexual isolation is caused by chemosensory cues. In our experiments, bacteria altered the profile of cuticular hydrocarbons, which function as sex pheromones. Commensal bacteria may act directly on these hydrocarbons, or they may affect their synthesis. Alternatively, bacterial metabolites might perfume the flies in ways that affect mate choice. In that event, habituation or conditioning likely plays a role.}, } @article {pmid21515811, year = {2011}, author = {Atyame, CM and Delsuc, F and Pasteur, N and Weill, M and Duron, O}, title = {Diversification of Wolbachia endosymbiont in the Culex pipiens mosquito.}, journal = {Molecular biology and evolution}, volume = {28}, number = {10}, pages = {2761-2772}, doi = {10.1093/molbev/msr083}, pmid = {21515811}, issn = {1537-1719}, mesh = {Animals ; Cluster Analysis ; Culex/genetics/*microbiology ; DNA, Mitochondrial/genetics ; Gene Rearrangement ; Genetic Markers ; Phylogeny ; Phylogeography ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {The α-proteobacteria Wolbachia are among the most common intracellular bacteria and have recently emerged as important drivers of arthropod biology. Wolbachia commonly act as reproductive parasites in arthropods by inducing cytoplasmic incompatibility (CI), a type of conditional sterility between hosts harboring incompatible infections. In this study, we examined the evolutionary histories of Wolbachia infections, known as wPip, in the common house mosquito Culex pipiens, which exhibits the greatest variation in CI crossing patterns observed in any insect. We first investigated a panel of 20 wPip strains for their genetic diversity through a multilocus scheme combining 13 Wolbachia genes. Because Wolbachia depend primarily on maternal transmission for spreading within arthropod populations, we also studied the variability in the coinherited Cx. pipiens mitochondria. In total, we identified 14 wPip haplotypes, which all share a monophyletic origin and clearly cluster into five distinct wPip groups. The diversity of Cx. pipiens mitochondria was extremely reduced, which is likely a consequence of cytoplasmic hitchhiking driven by a unique and recent Wolbachia invasion. Phylogenetic evidence indicates that wPip infections and mitochondrial DNA have codiverged through stable cotransmission within the cytoplasm and shows that a rapid diversification of wPip has occurred. The observed pattern demonstrates that a considerable degree of Wolbachia diversity can evolve within a single host species over short evolutionary periods. In addition, multiple signatures of recombination were found in most wPip genomic regions, leading us to conclude that the mosaic nature of wPip genomes may play a key role in their evolution.}, } @article {pmid21495472, year = {2011}, author = {Coffey, R}, title = {Outsmarting dengue fever.}, journal = {Scientific American}, volume = {304}, number = {4}, pages = {16}, doi = {10.1038/scientificamerican0411-16}, pmid = {21495472}, issn = {0036-8733}, mesh = {Aedes/*immunology/microbiology ; Animals ; Bacterial Vaccines/*administration & dosage ; Dengue/*prevention & control ; *Disease Vectors ; Humans ; Wolbachia/immunology ; }, } @article {pmid21481216, year = {2011}, author = {Simões, PM and Mialdea, G and Reiss, D and Sagot, MF and Charlat, S}, title = {Wolbachia detection: an assessment of standard PCR protocols.}, journal = {Molecular ecology resources}, volume = {11}, number = {3}, pages = {567-572}, doi = {10.1111/j.1755-0998.2010.02955.x}, pmid = {21481216}, issn = {1755-0998}, mesh = {Animals ; Arthropods/microbiology ; DNA Primers/genetics ; Entomology/*methods ; Nematoda/microbiology ; Polymerase Chain Reaction/*methods ; RNA, Ribosomal, 16S/genetics ; Sensitivity and Specificity ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia is a large monophyletic genus of intracellular bacteria, traditionally detected using PCR assays. Its considerable phylogenetic diversity and impact on arthropods and nematodes make it urgent to assess the efficiency of these screening protocols. The sensitivity and range of commonly used PCR primers and of a new set of 16S primers were evaluated on a wide range of hosts and Wolbachia strains. We show that certain primer sets are significantly more efficient than others but that no single protocol can ensure the specific detection of all known Wolbachia infections.}, } @article {pmid21476990, year = {2011}, author = {Thierry, M and Becker, N and Hajri, A and Reynaud, B and Lett, JM and Delatte, H}, title = {Symbiont diversity and non-random hybridization among indigenous (Ms) and invasive (B) biotypes of Bemisia tabaci.}, journal = {Molecular ecology}, volume = {20}, number = {10}, pages = {2172-2187}, doi = {10.1111/j.1365-294X.2011.05087.x}, pmid = {21476990}, issn = {1365-294X}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Enterobacteriaceae/classification/genetics/physiology ; Hemiptera/*microbiology ; Hybridization, Genetic/genetics ; Microsatellite Repeats/*genetics ; Phylogeny ; Rickettsia/classification/genetics/physiology ; Symbiosis/genetics ; Wolbachia/classification/genetics/physiology ; }, abstract = {The whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a worldwide pest and a vector of numerous plant viruses. B. tabaci is composed of dozens of morphologically indistinguishable biotypes and its taxonomic status is still controversial. This phloem-feeder harbours the primary symbiont Portiera aleyrodidarum and potentially six secondary symbionts: Cardinium, Arsenophonus, Hamiltonella, Rickettsia, Wolbachia and Fritschea. In the southwest Indian Ocean, La Réunion hosts two biotypes of this species: B (invasive) and Ms (indigenous). A multiplex PCR was developed to study the symbiont community of B. tabaci on La Réunion. Symbiont community prevalence and composition, host mitochondrial and nuclear genetic diversity, as well as host plant and localization, were described on field populations of La Réunion for B and Ms B. tabaci biotypes and their hybrids. A clear association between symbiotypes and biotypes was shown. Cardinium, Arsenophonus and Rickettsia were found in the Ms biotype (73.6%, 64.2% and 3.3%, respectively). Hamiltonella (exclusively) and Rickettsia were found in the B biotype (78% and 91.2%, respectively). Hybrids harboured all symbiotypes found in Ms and B populations, but with a higher prevalence of Ms symbiotypes than expected under random hybridization. An unexpected majority was Cardinium mono-infected (65.6%), and a striking minority (9%) harboured Cardinium/Arsenophonus. In the hybrids only, genetic diversity was linked to symbiotype. Among the hybrids, significant links were found between symbiotypes and: (i) mitochondrial COI sequences, i.e. maternal origin; and (ii) alleles of nuclear microsatellite loci, specific to either Ms or B parental biotype. Taken together, our results suggest that Cardinium and/or Arsenophonus may manipulate the reproduction of indigenous (Ms) with invasive (B) biotypes of Bemisia tabaci.}, } @article {pmid21460541, year = {2011}, author = {Hancock, PA and Sinkins, SP and Godfray, HC}, title = {Population dynamic models of the spread of Wolbachia.}, journal = {The American naturalist}, volume = {177}, number = {3}, pages = {323-333}, doi = {10.1086/658121}, pmid = {21460541}, issn = {1537-5323}, support = {079059/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animal Migration ; Animals ; Host-Pathogen Interactions ; Insect Vectors/microbiology ; Life Cycle Stages ; *Models, Biological ; Population Density ; Population Dynamics ; Seasons ; *Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia are endosymbionts that are found in many insect species and can spread rapidly when introduced into a naive host population. Most Wolbachia spread when their infection frequency exceeds a threshold normally calculated using purely population genetic models. However, spread may also depend on the population dynamics of the insect host. We develop models to explore interactions between host population dynamics and Wolbachia infection frequency for an age-structured insect population regulated by larval density dependence. We first derive a new expression for the threshold frequency that extends existing theory to incorporate important details of the insect's life history. In the presence of immigration and emigration, the threshold also depends on the form of density-dependent regulation. We show how the type of immigration (constant or pulsed) and the temporal dynamics of the host population can strongly affect the spread of Wolbachia. The results help understand the natural dynamics of Wolbachia infections and aid the design of programs to introduce Wolbachia to control insects that are disease vectors or pests.}, } @article {pmid21457543, year = {2011}, author = {Sun, XJ and Xiao, JH and Cook, JM and Feng, G and Huang, DW}, title = {Comparisons of host mitochondrial, nuclear and endosymbiont bacterial genes reveal cryptic fig wasp species and the effects of Wolbachia on host mtDNA evolution and diversity.}, journal = {BMC evolutionary biology}, volume = {11}, number = {}, pages = {86}, pmid = {21457543}, issn = {1471-2148}, mesh = {Animals ; Bacterial Proteins/*genetics ; DNA, Mitochondrial/*genetics ; *Evolution, Molecular ; Ficus ; *Genetic Variation ; Insect Proteins/genetics ; Molecular Sequence Data ; Phylogeny ; *Symbiosis ; Wasps/classification/*genetics/*microbiology/physiology ; Wolbachia/*genetics/physiology ; }, abstract = {BACKGROUND: Figs and fig-pollinating wasp species usually display a highly specific one-to-one association. However, more and more studies have revealed that the "one-to-one" rule has been broken. Co-pollinators have been reported, but we do not yet know how they evolve. They may evolve from insect speciation induced or facilitated by Wolbachia which can manipulate host reproduction and induce reproductive isolation. In addition, Wolbachia can affect host mitochondrial DNA evolution, because of the linkage between Wolbachia and associated mitochondrial haplotypes, and thus confound host phylogeny based on mtDNA. Previous research has shown that fig wasps have the highest incidence of Wolbachia infection in all insect taxa, and Wolbachia may have great influence on fig wasp biology. Therefore, we look forward to understanding the influence of Wolbachia on mitochondrial DNA evolution and speciation in fig wasps.

RESULTS: We surveyed 76 pollinator wasp specimens from nine Ficus microcarpa trees each growing at a different location in Hainan and Fujian Provinces, China. We found that all wasps were morphologically identified as Eupristina verticillata, but diverged into three clades with 4.22-5.28% mtDNA divergence and 2.29-20.72% nuclear gene divergence. We also found very strong concordance between E. verticillata clades and Wolbachia infection status, and the predicted effects of Wolbachia on both mtDNA diversity and evolution by decreasing mitochondrial haplotypes.

CONCLUSIONS: Our study reveals that the pollinating wasp E. verticillata on F. microcarpa has diverged into three cryptic species, and Wolbachia may have a role in this divergence. The results also indicate that Wolbachia strains infecting E. verticillata have likely resulted in selective sweeps on host mitochondrial DNA.}, } @article {pmid21457171, year = {2011}, author = {Adachi-Hagimori, T and Miura, K and Abe, Y}, title = {Gene flow between sexual and asexual strains of parasitic wasps: a possible case of sympatric speciation caused by a parthenogenesis-inducing bacterium.}, journal = {Journal of evolutionary biology}, volume = {24}, number = {6}, pages = {1254-1262}, doi = {10.1111/j.1420-9101.2011.02257.x}, pmid = {21457171}, issn = {1420-9101}, mesh = {Animals ; Female ; Fertilization/genetics/physiology ; *Gene Flow ; Genetic Speciation ; Genome ; Japan ; Male ; Parthenogenesis/*genetics ; Population Dynamics ; *Sexual Behavior, Animal ; Spermatozoa/physiology ; Wasps/*genetics/microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Sympatric speciation is strictly defined as the emergence of two species from a population in which mating has been random with respect to the place of birth of the mating partners. Mathematical models have shown that sympatric speciation is possible, but very few examples have been documented in nature. In this article, we demonstrate that arrhenotokous and thelytokous strains of a parasitic wasp, Neochrysocharis formosa, speciated sympatrically through infection by a symbiotic bacterium Rickettsia for the following reasons: First, Rickettsia infection was detected in all of the thelytokous strains collected throughout Japan. Second, the arrhenotokous and thelytokous strains have been collected sympatrically. Third, crossing experiments between the two strains did not result in fertilized offspring. In addition, the two strains were genetically isolated at the nuclear and mitochondrial genes. Fourth, the two strains showed a sister relationship in nuclear 28S rRNA gene. Finally, thelytokous females treated with antibiotics produced Rickettsia-free male offspring of the same reproductive form as arrhenotokous females indicating that the thelytokous strain could have speciated sympatrically from an individual of the arrhenotokous strain.}, } @article {pmid24031693, year = {2011}, author = {Marcon, HS and Coscrato, VE and Selivon, D and Perondini, AL and Marino, CL}, title = {Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae).}, journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]}, volume = {42}, number = {2}, pages = {778-785}, pmid = {24031693}, issn = {1517-8382}, abstract = {Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity.}, } @article {pmid23908883, year = {2011}, author = {Chang, MS and Christophel, EM and Gopinath, D and Abdur, RM and , and Vectorborne, O and Diseases, P and , and , }, title = {Challenges and future perspective for dengue vector control in the Western Pacific Region.}, journal = {Western Pacific surveillance and response journal : WPSAR}, volume = {2}, number = {2}, pages = {9-16}, pmid = {23908883}, issn = {2094-7321}, abstract = {Dengue remains a significant public health issue in the Western Pacific Region. In the absence of a vaccine, vector control is the mainstay for dengue prevention and control. In this paper we describe vector surveillance and vector control in the Western Pacific countries and areas. Vector surveillance and control strategies used by countries and areas of the Western Pacific Region vary. Vector control strategies include chemical, biological and environmental management that mainly target larval breeding sites. The use of insecticides targeting larvae and adult mosquitoes remains the mainstay of vector control programmes. Existing vector control tools have several limitations in terms of cost, delivery and long-term sustainability. However, there are several new innovative tools in the pipeline. These include Release of Insects Carrying a Dominant Lethal system and Wolbachia, an endosymbiotic bacterium, to inhibit dengue virus in the vector. In addition, the use of biological control such as larvivorous fish in combination with community participation has potential to be scaled up. Any vector control strategy should be selected based on evidence and appropriateness for the entomological and epidemiological setting and carried out in both inter-epidemic and epidemic periods. Community participation and interagency collaboration are required for effective and sustainable dengue prevention and control. Countries and areas are now moving towards integrated vector management.}, } @article {pmid21445333, year = {2011}, author = {Ritchie, SA and Johnson, PH and Freeman, AJ and Odell, RG and Graham, N and Dejong, PA and Standfield, GW and Sale, RW and O'Neill, SL}, title = {A secure semi-field system for the study of Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {5}, number = {3}, pages = {e988}, pmid = {21445333}, issn = {1935-2735}, mesh = {Aedes/*growth & development/*microbiology ; Animals ; Australia ; Containment of Biohazards/methods ; Entomology/*methods ; Female ; Male ; Mosquito Control/*methods ; Pest Control, Biological/methods ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: New contained semi-field cages are being developed and used to test novel vector control strategies of dengue and malaria vectors. We herein describe a new Quarantine Insectary Level-2 (QIC-2) laboratory and field cages (James Cook University Mosquito Research Facility Semi-Field System; MRF SFS) that are being used to measure the impact of the endosymbiont Wolbachia pipientis on populations of Aedes aegypti in Cairns Australia.

The MRF consists of a single QIC-2 laboratory/insectary that connects through a central corridor to two identical QIC-2 semi-field cages. The semi-field cages are constructed of two layers of 0.25 mm stainless steel wire mesh to prevent escape of mosquitoes and ingress of other insects. The cages are covered by an aluminum security mesh to prevent penetration of the cages by branches and other missiles in the advent of a tropical cyclone. Parts of the cage are protected from UV light and rainfall by 90% shade cloth and a vinyl cover. A wooden structure simulating the understory of a Queenslander-style house is also situated at one end of each cage. The remainder of the internal aspect of the cage is covered with mulch and potted plants to emulate a typical yard. An air conditioning system comprised of two external ACs that feed cooled, moistened air into the cage units. The air is released from the central ceiling beam from a long cloth tube that disperses the airflow and also prevents mosquitoes from escaping the cage via the AC system. Sensors located inside and outside the cage monitor ambient temperature and relative humidity, with AC controlled to match ambient conditions. Data loggers set in the cages and outside found a <2 °C temperature difference. Additional security features include air curtains over exit doors, sticky traps to monitor for escaping mosquitoes between layers of the mesh, a lockable vestibule leading from the connecting corridor to the cage and from inside to outside of the insectary, and screened (0.25 mm mesh) drains within the insectary and the cage. A set of standard operating procedures (SOP) has been developed to ensure that security is maintained and for enhanced surveillance for escaping mosquitoes on the JCU campus where the MRF is located. A cohort of male and female Aedes aegypti mosquitoes were released in the cage and sampled every 3-4 days to determine daily survival within the cage; log linear regression from BG-sentinel trapping collections produced an estimated daily survival of 0.93 and 0.78 for females and males, respectively.

CONCLUSIONS/SIGNIFICANCE: The MRF SFS allows us to test novel control strategies within a secure, contained environment. The air-conditioning system maintains conditions within the MRF cages comparable to outside ambient conditions. This cage provides a realistic transitional platform between the laboratory and the field in which to test novel control measures on quarantine level insects.}, } @article {pmid21410491, year = {2011}, author = {Zhang, X and Norris, DE and Rasgon, JL}, title = {Distribution and molecular characterization of Wolbachia endosymbionts and filarial nematodes in Maryland populations of the lone star tick (Amblyomma americanum).}, journal = {FEMS microbiology ecology}, volume = {77}, number = {1}, pages = {50-56}, pmid = {21410491}, issn = {1574-6941}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21AI067386/AI/NIAID NIH HHS/United States ; R21 AI067386/AI/NIAID NIH HHS/United States ; R03AI079297/AI/NIAID NIH HHS/United States ; R03 AI079297/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Female ; Filarioidea/classification/*genetics/isolation & purification ; Ixodidae/*microbiology/*parasitology ; Male ; Maryland ; Multilocus Sequence Typing ; Nematoda/microbiology ; Nymph/microbiology/parasitology ; Phylogeny ; Polymerase Chain Reaction ; *Symbiosis ; Wolbachia/classification/*genetics/isolation & purification ; }, abstract = {The lone star tick Amblyomma americanum is host to a wide diversity of endosymbiotic bacteria. We identified a novel Wolbachia symbiont infecting A. americanum. Multilocus sequence typing phylogenetically placed the endosymbiont in the increasingly diverse F supergroup. We assayed a total of 1031 ticks (119 females, 78 males and 834 nymphs in 89 pools) from 16 Maryland populations for infection. Infection frequencies in the natural populations were approximately 5% in females and <2% (minimum infection rate) in nymphs; infection was not detected in males. Infected populations were only observed in southern Maryland, suggesting the possibility that Wolbachia is currently invading Maryland A. americanum populations. Because F supergroup Wolbachia have been detected previously in filarial nematodes, tick samples were assayed for nematodes by PCR. Filarial nematodes were detected in 70% and 9% of Wolbachia-positive and Wolbachia-negative tick samples, respectively. While nematodes were more common in Wolbachia-positive tick samples, the lack of a strict infection concordance (Wolbachia-positive, nematode-negative and Wolbachia-negative, nematode-positive ticks) suggests that Wolbachia prevalence in ticks is not due to nematode infection. Supporting this hypothesis, phylogenetic analysis indicated that the nematodes were likely a novel species within the genus Acanthocheilonema, which has been previously shown to be Wolbachia-free.}, } @article {pmid21399578, year = {2010}, author = {Ravikumar, H and Ramachandraswamy, N and Sampathkumar, S and Prakash, BM and Huchesh, HC and Uday, J and Puttaraju, HP}, title = {A preliminary survey for Wolbachia and bacteriophage WO infections in Indian mosquitoes (Diptera: Culicidae).}, journal = {Tropical biomedicine}, volume = {27}, number = {3}, pages = {384-393}, pmid = {21399578}, issn = {2521-9855}, mesh = {Animals ; Bacteriophages/*isolation & purification ; Culicidae/*microbiology ; DNA, Bacterial/genetics ; Genes, Bacterial ; India ; Polymerase Chain Reaction ; Wolbachia/classification/genetics/*isolation & purification/*virology ; }, abstract = {Maternally inherited Wolbachia endosymbiotic bacteria are known to induce various kinds of reproductive alterations in their arthropod hosts. It has been proposed that this bacterium can be used as a tool for gene drive system in mosquitoes and also for the reduction of population size and modulating population age structure in order to reduce disease transmission. In the present study, we carried out a survey to determine the prevalence of Wolbachia and its phage WO infection in Indian mosquitoes and classified Wolbachia infection into groups A and B based on extensive polymerase chain reaction assay using Wolbachia specific wsp and orf7 gene primers. Out of 20 fieldcaught mosquito species, eight species have shown to be infected. Singly infected with Wolbachia A was found in two species and B group found in four species, while double infection with AB group were found in two species. All the screened mosquito species with positive Wolbachia infection were also infected with phage WO. The knowledge of variation in Wolbachia and phage WO infection rates and inferred susceptibility to infection among different mosquito genera has fundamental implications for designing and successful application of Wolbachia based vector-borne disease control strategies.}, } @article {pmid21398605, year = {2011}, author = {Specht, S and Frank, JK and Alferink, J and Dubben, B and Layland, LE and Denece, G and Bain, O and Förster, I and Kirschning, CJ and Martin, C and Hoerauf, A}, title = {CCL17 controls mast cells for the defense against filarial larval entry.}, journal = {Journal of immunology (Baltimore, Md. : 1950)}, volume = {186}, number = {8}, pages = {4845-4852}, doi = {10.4049/jimmunol.1000612}, pmid = {21398605}, issn = {1550-6606}, mesh = {Animals ; Antigens, Helminth/immunology ; Bone Marrow Cells/immunology/metabolism ; Capillary Permeability/immunology ; Cell Degranulation/immunology ; Cells, Cultured ; Chemokine CCL17/genetics/*immunology/metabolism ; Dendritic Cells/immunology/metabolism ; Female ; Filariasis/genetics/*immunology/parasitology ; Filarioidea/*immunology/microbiology/physiology ; Flow Cytometry ; Green Fluorescent Proteins/genetics/metabolism ; Host-Parasite Interactions/immunology ; Larva/immunology/microbiology/physiology ; Lung/immunology/metabolism ; Lymph Nodes/immunology/metabolism ; Mast Cells/*immunology/physiology ; Mice ; Mice, Inbred C3H ; Mice, Knockout ; Microscopy, Confocal ; Skin/immunology/metabolism ; Time Factors ; Wolbachia/immunology ; }, abstract = {Filarial parasites have to trespass many barriers to successfully settle within their mammalian host, which is equipped with mechanical borders and complex weaponry of an evolved immune system. However, little is known about mechanisms of early local events in filarial infections. In this study, bone marrow-derived dendritic cells not only upregulated activation markers CD40 and CD80 upon in vitro stimulation with filarial extracts, but also secreted CCL17, a chemokine known to be produced upon microbial challenge. Mice deficient for CCL17 had an up to 4-fold higher worm burden compared with controls by day 10 of infection with the murine filaria Litomosoides sigmodontis. Also, numbers of mast cells (MCs) invading the skin and degranulation were significantly increased, which was associated with enhanced vascular permeability and larval establishment. This phenotype was reverted by inhibition of MC degranulation with disodium cromoglycate or by blockade of histamine. In addition, we showed that CCL17-mediated vascular permeability was dependent on the presence of Wolbachia endosymbionts and TLR2. Our findings reveal that CCL17 controls filarial larval entry by limiting MC-dependent vascular permeability.}, } @article {pmid21379333, year = {2011}, author = {Hughes, GL and Ren, X and Ramirez, JL and Sakamoto, JM and Bailey, JA and Jedlicka, AE and Rasgon, JL}, title = {Wolbachia infections in Anopheles gambiae cells: transcriptomic characterization of a novel host-symbiont interaction.}, journal = {PLoS pathogens}, volume = {7}, number = {2}, pages = {e1001296}, pmid = {21379333}, issn = {1553-7374}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21 AI070178-01A1/AI/NIAID NIH HHS/United States ; R21 AI070178-02/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/genetics/*metabolism/*microbiology ; Biomarkers/metabolism ; Drosophila/genetics/microbiology ; Gene Expression Profiling ; Humans ; Malaria/*genetics/*microbiology ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Wolbachia/*metabolism/*pathogenicity ; }, abstract = {The endosymbiotic bacterium Wolbachia is being investigated as a potential control agent in several important vector insect species. Recent studies have shown that Wolbachia can protect the insect host against a wide variety of pathogens, resulting in reduced transmission of parasites and viruses. It has been proposed that compromised vector competence of Wolbachia-infected insects is due to up-regulation of the host innate immune system or metabolic competition. Anopheles mosquitoes, which transmit human malaria parasites, have never been found to harbor Wolbachia in nature. While transient somatic infections can be established in Anopheles, no stable artificially-transinfected Anopheles line has been developed despite numerous attempts. However, cultured Anopheles cells can be stably infected with multiple Wolbachia strains such as wAlbB from Aedes albopictus, wRi from Drosophila simulans and wMelPop from Drosophila melanogaster. Infected cell lines provide an amenable system to investigate Wolbachia-Anopheles interactions in the absence of an infected mosquito strain. We used Affymetrix GeneChip microarrays to investigate the effect of wAlbB and wRi infection on the transcriptome of cultured Anopheles Sua5B cells, and for a subset of genes used quantitative PCR to validate results in somatically-infected Anopheles mosquitoes. Wolbachia infection had a dramatic strain-specific effect on gene expression in this cell line, with almost 700 genes in total regulated representing a diverse array of functional classes. Very strikingly, infection resulted in a significant down-regulation of many immune, stress and detoxification-related transcripts. This is in stark contrast to the induction of immune genes observed in other insect hosts. We also identified genes that may be potentially involved in Wolbachia-induced reproductive and pathogenic phenotypes. Somatically-infected mosquitoes had similar responses to cultured cells. The data show that Wolbachia has a profound and unique effect on Anopheles gene expression in cultured cells, and has important implications for mechanistic understanding of Wolbachia-induced phenotypes and potential novel strategies to control malaria.}, } @article {pmid21364694, year = {2011}, author = {Christie, JS and Picornell, A and Moya, A and Ramon, MM and Castro, JA}, title = {Mitochondrial DNA effects on fitness in Drosophila subobscura.}, journal = {Heredity}, volume = {107}, number = {3}, pages = {239-245}, pmid = {21364694}, issn = {1365-2540}, mesh = {Animals ; Cell Nucleus/*genetics ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics ; Female ; Fertility/genetics ; Genetic Drift ; *Genetic Fitness ; Genetic Variation ; Haplotypes ; Hybrid Vigor ; Longevity/genetics ; Male ; Mutation ; Selection, Genetic ; Spain ; Wolbachia/isolation & purification ; }, abstract = {We tested different fitness components on a series of conspecific mtDNA haplotypes, detected by RFLPs in Drosophila subobscura. Additionally, haplotype VIII, endemic to the Canary Islands, was tested upon its own native nuclear DNA background and upon that of the rest of mtDNAs tested herein. We found that both nuclear and mitochondrial DNA can have a significant effect upon their hosts' fitness, and that negative selection is one of the mechanisms that can intervene in this species' mtDNA haplotype pattern. We discuss the importance of this mechanism in relation to genetic drift, in the form of periodic population bottlenecks, and how the latter can enhance the former. We also detected a significant positive effect of haplotype VIII upon fitness that could explain in part the dominance of this endemic haplotype on some of the Canary Islands, and a mitochondrial heterosis involving this haplotype when on a foreign nuclear DNA background.}, } @article {pmid21356077, year = {2011}, author = {Moreira, LA and Ye, YH and Turner, K and Eyles, DW and McGraw, EA and O'Neill, SL}, title = {The wMelPop strain of Wolbachia interferes with dopamine levels in Aedes aegypti.}, journal = {Parasites & vectors}, volume = {4}, number = {}, pages = {28}, pmid = {21356077}, issn = {1756-3305}, mesh = {Aedes/*chemistry/*microbiology/physiology ; Animals ; Biosynthetic Pathways ; Dopamine/*biosynthesis ; Female ; Gene Expression Profiling ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is an intracellular bacterium that has been stably transinfected into the mosquito vector of dengue, Aedes aegypti. This inherited infection causes a range of metabolic and phenotypic alterations in the mosquito, which might be related to neuronal abnormalities. In order to determine if these alterations were caused by the manipulation of neuroamines by this bacterium, we studied the expression of genes involved in the dopamine biosynthetic pathway and also measured the amount of dopamine in infected and uninfected mosquitoes of different ages. Wolbachia-infected mosquitoes exhibit greater expression of some genes related to the melanization pathway, but not for those directly linked to dopamine production. Although dopamine levels were higher in Wolbachia-positive mosquitoes this was not consistent across all insect ages nor was it related to the previously described Wolbachia induced "bendy" and "shaky" phenotypes.}, } @article {pmid21351390, year = {2011}, author = {Christodoulou, M}, title = {Biological vector control of mosquito-borne diseases.}, journal = {The Lancet. Infectious diseases}, volume = {11}, number = {2}, pages = {84-85}, doi = {10.1016/s1473-3099(11)70017-2}, pmid = {21351390}, issn = {1474-4457}, mesh = {Animals ; Animals, Genetically Modified/parasitology ; Culicidae/*microbiology/*parasitology ; Dengue/*prevention & control ; *Disease Vectors ; Female ; Humans ; Malaria, Falciparum/*prevention & control ; Male ; Mosquito Control/*methods ; Wolbachia/pathogenicity ; }, } @article {pmid21350051, year = {2011}, author = {Foster, J and Slatko, B and Bandi, C and Kumar, S}, title = {Recombination in wolbachia endosymbionts of filarial nematodes?.}, journal = {Applied and environmental microbiology}, volume = {77}, number = {5}, pages = {1921-1922}, pmid = {21350051}, issn = {1098-5336}, mesh = {Animals ; DNA, Bacterial/*genetics ; Filarioidea/*microbiology ; *Recombination, Genetic ; Sequence Alignment ; Symbiosis ; Wolbachia/*genetics/physiology ; }, } @article {pmid21349119, year = {2011}, author = {Nakamura, Y and Gotoh, T and Imanishi, S and Mita, K and Kurtti, TJ and Noda, H}, title = {Differentially expressed genes in silkworm cell cultures in response to infection by Wolbachia and Cardinium endosymbionts.}, journal = {Insect molecular biology}, volume = {20}, number = {3}, pages = {279-289}, doi = {10.1111/j.1365-2583.2010.01056.x}, pmid = {21349119}, issn = {1365-2583}, mesh = {Animals ; Antimicrobial Cationic Peptides/genetics ; *Bacteroidetes ; Bombyx/genetics/*immunology/*microbiology ; Cells, Cultured ; Cytoplasm/immunology/microbiology ; *Gene Expression Regulation ; Immunity/*genetics ; Oligonucleotide Array Sequence Analysis ; Symbiosis/genetics/*immunology ; *Wolbachia ; }, abstract = {Wolbachia and Cardinium are bacterial endosymbionts that are widely distributed amongst arthropods. Both cause reproductive alterations, such as cytoplasmic incompatibility, parthenogenesis and feminization. Here we studied differentially expressed genes in Wolbachia- and Cardinium-infected Bm-aff3 silkworm cells using a silkworm microarray. Wolbachia infection did not alter gene expression or induce or suppress immune responses. In contrast, Cardinium infection induced many immune-related genes, including antimicrobial peptides, pattern recognition receptors and a serine protease. Host immune responses differed, possibly because of the different cell wall structures of Wolbachia and Cardinium because the former lacks genes encoding lipopolysaccharide components and two racemases for peptidoglycan formation. A few possibly non-immune-related genes were differentially expressed, but their involvement in host reproductive alteration was unclear.}, } @article {pmid21337952, year = {2011}, author = {McMeniman, CJ and Hughes, GL and O'Neill, SL}, title = {A Wolbachia symbiont in Aedes aegypti disrupts mosquito egg development to a greater extent when mosquitoes feed on nonhuman versus human blood.}, journal = {Journal of medical entomology}, volume = {48}, number = {1}, pages = {76-84}, doi = {10.1603/me09188}, pmid = {21337952}, issn = {0022-2585}, mesh = {Aedes/*microbiology/physiology ; Animals ; Feeding Behavior ; Female ; Guinea Pigs ; Humans ; Mice ; *Oviparity ; Ovum/growth & development ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {A vertebrate bloodmeal is required by female mosquitoes of most species to obtain nutrients for egg maturation. The yellowfever mosquito, Aedes aegypti (L.), feeds predominantly on humans, despite having the capacity to use blood from other hosts for this process. Here, we report that female Ae. aegypti infected with a virulent strain of the intracellular bacterium Wolbachia pipientis (wMelPop) from Drosophila melanogaster (Meigen) have a reduced ability to use blood for egg development. Blood feeding by wMelPop-infected females on mouse, guinea pig, or chicken hosts resulted in a near complete abolishment of reproductive output associated with both a decline in the numbers of eggs oviposited as well as the hatching rate of successfully laid eggs. In contrast, the reproductive output of wMelPop-infected females fed human blood was only mildly affected in comparison to individuals fed animal blood sources. Blood-feeding assays over two reproductive cycles definitively illustrated a nutritional interaction between host blood source and egg development in wMelPop-infected Ae. aegypti. Removal of Wolbachia from mosquitoes using antibiotic treatment rescued egg development on all blood sources. Further investigation of this phenotype may provide new insights into the nutritional basis of mosquito anthropophily.}, } @article {pmid21329550, year = {2011}, author = {Chu, D and Gao, CS and De Barro, P and Zhang, YJ and Wan, FH and Khan, IA}, title = {Further insights into the strange role of bacterial endosymbionts in whitefly, Bemisia tabaci: comparison of secondary symbionts from biotypes B and Q in China.}, journal = {Bulletin of entomological research}, volume = {101}, number = {4}, pages = {477-486}, doi = {10.1017/S0007485311000083}, pmid = {21329550}, issn = {1475-2670}, mesh = {Animals ; Bacterial Typing Techniques ; China ; Hemiptera/*microbiology ; Introduced Species ; Mediterranean Region ; Rickettsia/classification/isolation & purification ; *Symbiosis ; Wolbachia/classification/isolation & purification ; }, abstract = {The percentage infection of secondary symbionts (SS) (Wolbachia, Arsenophonus, Rickettsia, Hamiltonella, Fritschea and Cardinium) in the exotic Bemisia tabaci (Genn.) invaders, commonly known as biotypes B and Q from China, were determined by PCR. In total, 373 biotype B and 1830 biotype Q individuals were screened for the presence of SS. Biotype B was more abundant than biotype Q from 2005 to 2006, and biotype Q was more abundant from 2007 to 2009. Each of the SS, with the exception of Fritschea, was detected in both biotypes B and Q; Fritschea was found in none of the samples examined. For biotype B, the percentage infection of Hamiltonella was the highest (92.0%) followed by Rickettsia (70.2%). For biotype Q, the percentage infection of Hamiltonella was again the highest (73.3%). Arsenophonus was the least common of the SS observed in both biotypes B and Q. The percentage infection of Wolbachia, Rickettsia and Hamiltonella in biotype B was each significantly higher than in biotype Q, whereas the percentage infection of Cardinium in biotype B was significantly lower than in biotype Q. The percentage infection of SS in biotypes B and Q varied from year to year over the period 2005-2009. Furthermore, within biotype Q, two distinct subgroups were identified which differ from each other in terms of their SS complement. We discuss these results in the light of the potentially influential factors and roles of the SS.}, } @article {pmid21327558, year = {2011}, author = {Nachappa, P and Levy, J and Pierson, E and Tamborindeguy, C}, title = {Diversity of endosymbionts in the potato psyllid, Bactericera cockerelli (Triozidae), vector of zebra chip disease of potato.}, journal = {Current microbiology}, volume = {62}, number = {5}, pages = {1510-1520}, pmid = {21327558}, issn = {1432-0991}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; *Biodiversity ; Female ; Hemiptera/*microbiology/physiology ; Insect Vectors/*microbiology ; Male ; Molecular Sequence Data ; Phylogeny ; Plant Diseases/*microbiology ; Rhizobiaceae/classification/genetics/isolation & purification/physiology ; Solanum tuberosum/*microbiology/parasitology ; *Symbiosis ; }, abstract = {Zebra chip disease is an emerging, serious disease of solanaceous crops and the causal agent is a bacterium "Candidatus Liberibacter solanacearum" (CLs), also known as "Candidatus Liberibacter psyllaurous", which is transmitted by the potato psyllid, Bactericera cockerelli (Šulc). We performed bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP) of the 16S rDNA genes to determine the bacterial microbiota in adult insects from CLs-uninfected and CLs-infected strains of B. cockerelli and potato leaf samples. We obtained sequences from five bacterial species among the two psyllid strains, including "Candidatus Carsonella ruddii", Wolbachia, CLs, and two transient bacteria, Acinetobacter and Methylibium. We did not detect any common bacteria between psyllids and potato leaf samples using pyrosequencing. We performed PCR analysis using species-specific 16S rDNA primers to confirm pyrosequencing results in individual psyllids including eggs, early-instars, late-instars, and adults of both sexes from both CLs-uninfected and CLs-infected psyllid strains. The primary endosymbiont, "Candidatus Carsonella ruddii" and Wolbachia were detected in all life-stages and sexes of both strains using PCR analyses. The percentage of CLs-infected individuals increased from early-instar (0%), late-instar (40%) until adulthood (60%) in the CLs-infected strain. We believe that CLs levels in early-instars are probably too low to be detected by standard PCR. Using PCR analyses, we confirmed the presence of Acinetobacter in CLs-uninfected and CLs-infected adults (75 and 25%, respectively) but not Methylibium. Further, we detected Acinetobacter in potato leaves using PCR indicating that the psyllids may have acquired this bacterium via feeding on the host plant.}, } @article {pmid21324604, year = {2011}, author = {Li, Z and Galvin, BD and Raverdy, S and Carlow, CK}, title = {Identification and characterization of the cofactor-independent phosphoglycerate mutases of Dirofilaria immitis and its Wolbachia endosymbiont.}, journal = {Veterinary parasitology}, volume = {176}, number = {4}, pages = {350-356}, doi = {10.1016/j.vetpar.2011.01.020}, pmid = {21324604}, issn = {1873-2550}, support = {AI061865/AI/NIAID NIH HHS/United States ; }, mesh = {2,3-Diphosphoglycerate/metabolism ; Amino Acid Sequence ; Animals ; Bacterial Proteins/genetics/isolation & purification/metabolism ; Cloning, Molecular ; DNA, Complementary/chemistry/isolation & purification ; Dirofilaria immitis/*enzymology/genetics/microbiology ; Female ; Gene Expression ; Glyceric Acids/metabolism ; Helminth Proteins/genetics/isolation & purification/metabolism ; Molecular Sequence Data ; Phosphoglycerate Mutase/chemistry/genetics/isolation & purification/*metabolism ; Phylogeny ; Recombinant Proteins/genetics/isolation & purification/metabolism ; Sequence Alignment ; Symbiosis ; Wolbachia/*enzymology/genetics/physiology ; }, abstract = {Drug treatments for heartworm disease have not changed significantly in the last decade. Due to concerns about possible drug resistance and their lower efficacy against adult worms, there is a need for the development of new antifilarial drug therapies. The recent availability of genomic sequences for the related filarial parasite Brugia malayi and its Wolbachia endosymbiont enables genome-wide searching for new drug targets. Phosphoglycerate mutase (PGM) enzymes catalyze the critical isomerization of 3-phosphoglycerate (3-PG) and 2-phosphoglycerate (2-PG) in glycolytic and gluconeogenic metabolic pathways. There are two unrelated PGM enzymes, which are structurally distinct and possess different mechanisms of action. The mammalian enzyme requires 2,3-bisphosphoglycerate as a cofactor (dependent PGM or dPGM), while the other type of PGM does not (independent PGM or iPGM). In the present study, we have determined that Dirofilaria immitis and its Wolbachia endosymbiont both possess active iPGM. We describe the molecular characterization and catalytic properties of each enzyme. Our results will facilitate the discovery of selective inhibitors of these iPGMs as potentially novel drug treatments for heartworm disease.}, } @article {pmid21318277, year = {2011}, author = {Ahantarig, A and Malaisri, P and Hirunkanokpun, S and Sumrandee, C and Trinachartvanit, W and Baimai, V}, title = {Detection of Rickettsia and a novel Haemaphysalis shimoga symbiont bacterium in ticks in Thailand.}, journal = {Current microbiology}, volume = {62}, number = {5}, pages = {1496-1502}, pmid = {21318277}, issn = {1432-0991}, mesh = {Animals ; Female ; Ixodidae/*classification/genetics/*microbiology/physiology ; Male ; Molecular Sequence Data ; Phylogeny ; Rickettsia/classification/genetics/*isolation & purification/*physiology ; *Symbiosis ; Thailand ; }, abstract = {In this study, we identified two Haemaphysalis species present at the Khao Yai National Park in Thailand and investigated the presence of rickettsia in these ticks. A total of 166 Haemaphysalis specimens were collected randomly under leaves along visitor paths at five locations in the park. Male and female adults of two different Haemaphysalis species, H. shimoga and H. lagrangei, were identified. Polymerase chain reaction (PCR) analysis revealed Rickettsia bacteria in these two Haemaphysalis species; this study represents the first time such presence has been reported in Thailand. The infection rates of Rickettsia were in both H. shimoga (7.41%) and H. lagrangei (10.17%) at these locations in addition to two pools of Haemahysalis nymphs (28.57%). Furthermore, 25.93% of H. shimoga showed positive results that matched Haemaphysalis longicornis symbionts (92% sequence identity) and the Coxeilla burnetti 16S ribosomal RNA gene (90% sequence identity). We propose that this is a novel H. shimoga symbiont bacterium in Thailand and might be a novel Coxeilla-like agent or Coxeilla sp. found in H. shimoga. In contrast, we did not observe any Wolbachia bacteria, which also belong to the order Rickettsiales, in the same group of Haemaphysalis ticks. Furthermore, PCR was used to detect three other genera of bacteria, Anaplasma, Ehrlichia and Borrelia, none of which were identified in the Haemaphysalis ticks studied.}, } @article {pmid21292630, year = {2011}, author = {Kent, BN and Salichos, L and Gibbons, JG and Rokas, A and Newton, IL and Clark, ME and Bordenstein, SR}, title = {Complete bacteriophage transfer in a bacterial endosymbiont (Wolbachia) determined by targeted genome capture.}, journal = {Genome biology and evolution}, volume = {3}, number = {}, pages = {209-218}, pmid = {21292630}, issn = {1759-6653}, support = {F31 AI091343/AI/NIAID NIH HHS/United States ; R01 GM085163/GM/NIGMS NIH HHS/United States ; R24 GM084917/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Bacteriophages/*genetics/physiology ; *Gene Transfer, Horizontal ; *Genome, Bacterial ; Symbiosis ; Wolbachia/*genetics/physiology/*virology ; }, abstract = {Bacteriophage flux can cause the majority of genetic diversity in free-living bacteria. This tenet of bacterial genome evolution generally does not extend to obligate intracellular bacteria owing to their reduced contact with other microbes and a predominance of gene deletion over gene transfer. However, recent studies suggest intracellular coinfections in the same host can facilitate exchange of mobile elements between obligate intracellular bacteria-a means by which these bacteria can partially mitigate the reductive forces of the intracellular lifestyle. To test whether bacteriophages transfer as single genes or larger regions between coinfections, we sequenced the genome of the obligate intracellular Wolbachia strain wVitB from the parasitic wasp Nasonia vitripennis and compared it against the prophage sequences of the divergent wVitA coinfection. We applied, for the first time, a targeted sequence capture array to specifically trap the symbiont's DNA from a heterogeneous mixture of eukaryotic, bacterial, and viral DNA. The tiled array successfully captured the genome with 98.3% efficiency. Examination of the genome sequence revealed the largest transfer of bacteriophage and flanking genes (52.2 kb) to date between two obligate intracellular coinfections. The mobile element transfer occurred in the recent evolutionary past based on the 99.9% average nucleotide identity of the phage sequences between the two strains. In addition to discovering an evolutionary recent and large-scale horizontal phage transfer between coinfecting obligate intracellular bacteria, we demonstrate that "targeted genome capture" can enrich target DNA to alleviate the problem of isolating symbiotic microbes that are difficult to culture or purify from the conglomerate of organisms inside eukaryotes.}, } @article {pmid21292403, year = {2011}, author = {Kramer, L and Grandi, G and Passeri, B and Gianelli, P and Genchi, M and Dzimianski, MT and Supakorndej, P and Mansour, AM and Supakorndej, N and McCall, SD and McCall, JW}, title = {Evaluation of lung pathology in Dirofilaria immitis-experimentally infected dogs treated with doxycycline or a combination of doxycycline and ivermectin before administration of melarsomine dihydrochloride.}, journal = {Veterinary parasitology}, volume = {176}, number = {4}, pages = {357-360}, doi = {10.1016/j.vetpar.2011.01.021}, pmid = {21292403}, issn = {1873-2550}, mesh = {Administration, Oral ; Animals ; Anti-Bacterial Agents/administration & dosage/*therapeutic use ; Antiparasitic Agents/administration & dosage/*therapeutic use ; Arsenicals/administration & dosage/therapeutic use ; Dirofilaria immitis/drug effects/pathogenicity ; Dirofilariasis/*drug therapy/pathology ; Dog Diseases/*drug therapy/parasitology/pathology ; Dogs ; Doxycycline/administration & dosage/therapeutic use ; Drug Therapy, Combination/veterinary ; Female ; Injections, Intramuscular/veterinary ; Ivermectin/administration & dosage/therapeutic use ; Lung/*pathology ; Male ; Time Factors ; Triazines/administration & dosage/therapeutic use ; Wolbachia/drug effects/pathogenicity ; }, abstract = {Adulticide therapy in heartworm (Dirofilaria immitis)-infected dogs can lead to thromboembolism, which can seriously compromise post-treatment health status. Lung pathology following adulticide therapy was evaluated in three groups of experimentally infected dogs. Group 1 was treated with doxycycline at 20 mg/kg per os once daily for 30 days post infection followed by an intramuscular injection of melarsomine dihydrochloride (2.5 mg/kg) at Week 12, followed 1 month later by two injections 24 h apart. Group 2 was treated as described for Group 1, with the addition of ivermectin at 6 mcg/kg given monthly per os for 24 weeks post-infection. Group 3 received melarsomine alone, as described above. All dogs were necropsied at Week 24 and lung pathology was evaluated. Lesion criteria included perivascular inflammation and endothelial proliferation. Lesions were scored by two independent pathologists who were blinded as to treatment. Results indicate that doxycycline treatment alone or combined with ivermectin had lower lesion scores than lungs from dogs who had received melarsomine alone. Dogs that received the combined doxycycline/ivermectin protocol and treated with adulticide showed less severe arterial lesions and the virtual absence of thrombi.}, } @article {pmid21283830, year = {2011}, author = {Bai, X and Mamidala, P and Rajarapu, SP and Jones, SC and Mittapalli, O}, title = {Transcriptomics of the bed bug (Cimex lectularius).}, journal = {PloS one}, volume = {6}, number = {1}, pages = {e16336}, pmid = {21283830}, issn = {1932-6203}, mesh = {Animals ; Bedbugs/*genetics ; Ectoparasitic Infestations ; Expressed Sequence Tags ; Gene Expression Profiling/*methods ; Insecticide Resistance/*genetics ; Insecticides/pharmacology ; Microsatellite Repeats ; Polymorphism, Single Nucleotide ; RNA, Messenger/*analysis ; Sequence Analysis ; United States ; }, abstract = {BACKGROUND: Bed bugs (Cimex lectularius) are blood-feeding insects poised to become one of the major pests in households throughout the United States. Resistance of C. lectularius to insecticides/pesticides is one factor thought to be involved in its sudden resurgence. Despite its high-impact status, scant knowledge exists at the genomic level for C. lectularius. Hence, we subjected the C. lectularius transcriptome to 454 pyrosequencing in order to identify potential genes involved in pesticide resistance.

Using 454 pyrosequencing, we obtained a total of 216,419 reads with 79,596,412 bp, which were assembled into 35,646 expressed sequence tags (3902 contigs and 31744 singletons). Nearly 85.9% of the C. lectularius sequences showed similarity to insect sequences, but 44.8% of the deduced proteins of C. lectularius did not show similarity with sequences in the GenBank non-redundant database. KEGG analysis revealed putative members of several detoxification pathways involved in pesticide resistance. Lamprin domains, Protein Kinase domains, Protein Tyrosine Kinase domains and cytochrome P450 domains were among the top Pfam domains predicted for the C. lectularius sequences. An initial assessment of putative defense genes, including a cytochrome P450 and a glutathione-S-transferase (GST), revealed high transcript levels for the cytochrome P450 (CYP9) in pesticide-exposed versus pesticide-susceptible C. lectularius populations. A significant number of single nucleotide polymorphisms (296) and microsatellite loci (370) were predicted in the C. lectularius sequences. Furthermore, 59 putative sequences of Wolbachia were retrieved from the database.

CONCLUSIONS: To our knowledge this is the first study to elucidate the genetic makeup of C. lectularius. This pyrosequencing effort provides clues to the identification of potential detoxification genes involved in pesticide resistance of C. lectularius and lays the foundation for future functional genomics studies.}, } @article {pmid21283625, year = {2011}, author = {Friberg, U and Miller, PM and Stewart, AD and Rice, WR}, title = {Mechanisms promoting the long-term persistence of a Wolbachia infection in a laboratory-adapted population of Drosophila melanogaster.}, journal = {PloS one}, volume = {6}, number = {1}, pages = {e16448}, pmid = {21283625}, issn = {1932-6203}, support = {R01 HD057974/HD/NICHD NIH HHS/United States ; 1R01HD057974-01/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; Clinical Laboratory Techniques ; Cytoplasm ; Drosophila melanogaster/*microbiology ; Female ; Host Specificity/*physiology ; Insecta/microbiology ; Male ; Phenotype ; Rickettsiaceae Infections ; Symbiosis/*physiology ; *Wolbachia ; }, abstract = {Intracellular bacteria of the genus Wolbachia are widespread endosymbionts across diverse insect taxa. Despite this prevalence, our understanding of how Wolbachia persists within populations is not well understood. Cytoplasmic incompatibility (CI) appears to be an important phenotype maintaining Wolbachia in many insects, but it is believed to be too weak to maintain Wolbachia in Drosophila melanogaster, suggesting that Wolbachia must also have other effects on this species. Here we estimate the net selective effect of Wolbachia on its host in a laboratory-adapted population of D. melanogaster, to determine the mechanisms leading to its persistence in the laboratory environment. We found i) no significant effects of Wolbachia infection on female egg-to-adult survival or adult fitness, ii) no reduced juvenile survival in males, iii) substantial levels of CI, and iv) a vertical transmission rate of Wolbachia higher than 99%. The fitness of cured females was, however, severely reduced (a decline of 37%) due to CI in offspring. Taken together these findings indicate that Wolbachia is maintained in our laboratory environment due to a combination of a nearly perfect transmission rate and substantial CI. Our results show that there would be strong selection against females losing their infection and producing progeny free from Wolbachia.}, } @article {pmid21278277, year = {2011}, author = {Hughes, GL and Allsopp, PG and Brumbley, SM and Woolfit, M and McGraw, EA and O'Neill, SL}, title = {Variable infection frequency and high diversity of multiple strains of Wolbachia pipientis in Perkinsiella Planthoppers.}, journal = {Applied and environmental microbiology}, volume = {77}, number = {6}, pages = {2165-2168}, pmid = {21278277}, issn = {1098-5336}, mesh = {Molecular Sequence Data ; Phylogeny ; Saccharum/*microbiology ; Wolbachia/classification/*genetics ; }, abstract = {This survey of Wolbachia infections in populations of the planthoppers Perkinsiella saccharicida and Perkinsiella vitiensis revealed variable frequencies, low-titer infections, and high phylogenetic diversities of strains. These observations add to the growing realization that Wolbachia infections may be extremely common within invertebrates and yet occur infrequently within populations and at low titer within individuals.}, } @article {pmid21255170, year = {2011}, author = {Branca, A and LE Ru, BP and Vavre, F and Silvain, JF and Dupas, S}, title = {Intraspecific specialization of the generalist parasitoid Cotesia sesamiae revealed by polyDNAvirus polymorphism and associated with different Wolbachia infection.}, journal = {Molecular ecology}, volume = {20}, number = {5}, pages = {959-971}, doi = {10.1111/j.1365-294X.2010.04977.x}, pmid = {21255170}, issn = {1365-294X}, mesh = {Africa South of the Sahara ; Animals ; DNA, Viral/genetics ; Genes, Viral ; Genome, Insect ; Genotype ; Host-Parasite Interactions ; *Host-Pathogen Interactions ; Larva/parasitology ; Lepidoptera/parasitology ; Polydnaviridae/*genetics ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Wasps/genetics/*microbiology/*virology ; Wolbachia/genetics/*physiology ; }, abstract = {As a result of an intense host-parasite evolutionary arms race, parasitic wasps frequently display high levels of specialization on very few host species. For instance, in braconid wasps very few generalist species have been described. However, within this family, Cotesia sesamiae is a generalist species that is widespread in sub-Saharan Africa and develops on several lepidopteran hosts. In this study, we tested the hypothesis that C. sesamiae may be a cryptic specialist when examined at the intraspecific level. We sequenced exon 2 of CrV1, a gene of the symbiotic polyDNAvirus that is integrated into the wasp genome and is associated with host immune suppression. We found that CrV1 genotype was more closely associated with the host in which the parasitoid developed than any abiotic environmental factor tested. We also tested a correlation between CrV1 genotype and an infection with Wolbachia bacteria, which are known for their ability to induce reproductive isolation. The Wolbachia bacteria infection polymorphism was also found as a major factor explaining the genetic structure of CrV1, and, in addition, the best model explaining CrV1 genetic structure involved an interaction between Wolbachia infection and host species. We suggest that Wolbachia could act as an agent capable of maintaining advantageous alleles for host specialization in different populations of C. sesamiae. This mechanism could be applicable to other insect models because of the high prevalence of Wolbachia in insects.}, } @article {pmid21243351, year = {2011}, author = {Ishak, HD and Plowes, R and Sen, R and Kellner, K and Meyer, E and Estrada, DA and Dowd, SE and Mueller, UG}, title = {Bacterial diversity in Solenopsis invicta and Solenopsis geminata ant colonies characterized by 16S amplicon 454 pyrosequencing.}, journal = {Microbial ecology}, volume = {61}, number = {4}, pages = {821-831}, pmid = {21243351}, issn = {1432-184X}, mesh = {Animals ; Ants/*microbiology ; Bacteria/classification/genetics/*isolation & purification ; *Biodiversity ; DNA, Bacterial/*genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Sequence Analysis, DNA ; Species Specificity ; }, abstract = {Social insects harbor diverse assemblages of bacterial microbes, which may play a crucial role in the success or failure of biological invasions. The invasive fire ant Solenopsis invicta (Formicidae, Hymenoptera) is a model system for understanding the dynamics of invasive social insects and their biological control. However, little is known about microbes as biotic factors influencing the success or failure of ant invasions. This pilot study is the first attempt to characterize and compare microbial communities associated with the introduced S. invicta and the native Solenopsis geminata in the USA. Using 16S amplicon 454 pyrosequencing, bacterial communities of workers, brood, and soil from nest walls were compared between neighboring S. invicta and S. geminata colonies at Brackenridge Field Laboratory, Austin, Texas, with the aim of identifying potential pathogenic, commensal, or mutualistic microbial associates. Two samples of S. geminata workers showed high counts of Spiroplasma bacteria, a known pathogen or mutualist of other insects. A subsequent analysis using PCR and sequencing confirmed the presence of Spiroplasma in additional colonies of both Solenopsis species. Wolbachia was found in one alate sample of S. geminata, while one brood sample of S. invicta had a high count of Lactococcus. As expected, ant samples from both species showed much lower microbial diversity than the surrounding soil. Both ant species had similar overall bacterial diversities, although little overlap in specific microbes. To properly characterize a single bacterial community associated with a Solenopsis ant sample, rarefaction analyses indicate that it is necessary to obtain 5,000-10,000 sequences. Overall, 16S amplicon 454 pyrosequencing appears to be a cost-effective approach to screen whole microbial diversity associated with invasive ant species.}, } @article {pmid21235755, year = {2011}, author = {Vanthournout, B and Swaegers, J and Hendrickx, F}, title = {Spiders do not escape reproductive manipulations by Wolbachia.}, journal = {BMC evolutionary biology}, volume = {11}, number = {}, pages = {15}, pmid = {21235755}, issn = {1471-2148}, mesh = {Animals ; Bacteroidetes/classification/genetics/physiology ; Female ; Male ; Molecular Sequence Data ; Phylogeny ; Reproduction ; Rickettsia/classification/genetics/physiology ; Species Specificity ; Spiders/*microbiology/*physiology ; Symbiosis ; Wolbachia/classification/genetics/*physiology ; }, abstract = {BACKGROUND: Maternally inherited bacteria that reside obligatorily or facultatively in arthropods can increase their prevalence in the population by altering their hosts' reproduction. Such reproductive manipulations have been reported from the major arthropod groups such as insects (in particular hymenopterans, butterflies, dipterans and beetles), crustaceans (isopods) and mites. Despite the observation that endosymbiont bacteria are frequently encountered in spiders and that the sex ratio of particular spider species is strongly female biased, a direct relationship between bacterial infection and sex ratio variation has not yet been demonstrated for this arthropod order.

RESULTS: Females of the dwarf spider Oedothorax gibbosus exhibit considerable variation in the sex ratio of their clutches and were infected with at least three different endosymbiont bacteria capable of altering host reproduction i.e. Wolbachia, Rickettsia and Cardinium. Breeding experiments show that sex ratio variation in this species is primarily maternally inherited and that removal of the bacteria by antibiotics restores an unbiased sex ratio. Moreover, clutches of females infected with Wolbachia were significantly female biased while uninfected females showed an even sex ratio. As female biased clutches were of significantly smaller size compared to non-distorted clutches, killing of male embryos appears to be the most likely manipulative effect.

CONCLUSIONS: This represents to our knowledge the first direct evidence that endosymbiont bacteria, and in particular Wolbachia, might induce sex ratio variation in spiders. These findings are pivotal to further understand the diversity of reproductive phenotypes observed in this arthropod order.}, } @article {pmid21234752, year = {2011}, author = {Tabata, J and Hattori, Y and Sakamoto, H and Yukuhiro, F and Fujii, T and Kugimiya, S and Mochizuki, A and Ishikawa, Y and Kageyama, D}, title = {Male killing and incomplete inheritance of a novel spiroplasma in the moth Ostrinia zaguliaevi.}, journal = {Microbial ecology}, volume = {61}, number = {2}, pages = {254-263}, pmid = {21234752}, issn = {1432-184X}, mesh = {Animals ; Female ; Genes, Bacterial ; *Inheritance Patterns ; Male ; Moths/*microbiology ; Ovary/microbiology/ultrastructure ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Sex Ratio ; Spiroplasma/classification/genetics/*pathogenicity ; }, abstract = {Bacteria of the genus Spiroplasma are widely found in plants and arthropods. Some of the maternally transmitted Spiroplasma endosymbionts in arthropods are known to kill young male hosts (male killing). Here, we describe a new case of Spiroplasma-induced male killing in a moth, Ostrinia zaguliaevi. The all-female trait caused by Spiroplasma was maternally inherited for more than 11 generations but was spontaneously lost in several lineages. Antibiotic treatment eliminated the Spiroplasma infection and restored the 1:1 sex ratio. The survival rates and presence/absence of the W chromosome in the embryonic and larval stages of O. zaguliaevi showed that males were selectively killed, exclusively during late embryogenesis in all-female broods. Based on phylogenetic analyses of 16S rRNA, dnaA and rpoB gene sequences, the causative bacteria were identified as Spiroplasma belonging to the tick symbiont Spiroplasma ixodetis clade. Electron microscopy confirmed bacterial structures in the follicle cells and follicular sheath of adult females. Although many congeneric Ostrinia moths harbor another sex ratio-distorting bacterium (Wolbachia), only O. zaguliaevi harbors Spiroplasma.}, } @article {pmid21234244, year = {2011}, author = {Fernando, SD and Rodrigo, C and Rajapakse, S}, title = {Current evidence on the use of antifilarial agents in the management of bancroftian filariasis.}, journal = {Journal of tropical medicine}, volume = {2011}, number = {}, pages = {175941}, pmid = {21234244}, issn = {1687-9694}, abstract = {Many trials have explored the efficacy of individual drugs and drug combinations to treat bancroftian filariasis. This narrative review summarizes the current evidence for drug management of bancroftian filariasis. Diethylcarbamazine (DEC) remains the prime antifilarial agent with a well-established microfilaricidal and some macrofilaricidal effects. Ivermectin (IVM) is highly microfilaricidal but minimally macrofilaricidal. The role of albendazole (ALB) in treatment regimens is not well established though the drug has a microfilaricidal effect. The combination of DEC+ALB has a better long-term impact than IVM+ALB. Recent trials have shown that doxycycline therapy against Wolbachia, an endosymbiotic bacterium of the parasite, is capable of reducing microfilaria rates and adult worm activity. Followup studies on mass drug administration (MDA) are yet to show a complete interruption of transmission, though the infection rates are reduced to a very low level.}, } @article {pmid21229263, year = {2011}, author = {Raghavendra, K and Barik, TK and Reddy, BP and Sharma, P and Dash, AP}, title = {Malaria vector control: from past to future.}, journal = {Parasitology research}, volume = {108}, number = {4}, pages = {757-779}, pmid = {21229263}, issn = {1432-1955}, mesh = {Animals ; Drug Resistance ; History, 20th Century ; History, 21st Century ; Insecticides/pharmacology ; Malaria/*epidemiology/*prevention & control/transmission ; Mosquito Control/*history/*methods ; Pest Control, Biological/methods ; }, abstract = {Malaria is one of the most common vector-borne diseases widespread in the tropical and subtropical regions. Despite considerable success of malaria control programs in the past, malaria still continues as a major public health problem in several countries. Vector control is an essential part for reducing malaria transmission and became less effective in recent years, due to many technical and administrative reasons, including poor or no adoption of alternative tools. Of the different strategies available for vector control, the most successful are indoor residual spraying and insecticide-treated nets (ITNs), including long-lasting ITNs and materials. Earlier DDT spray has shown spectacular success in decimating disease vectors but resulted in development of insecticide resistance, and to control the resistant mosquitoes, organophosphates, carbamates, and synthetic pyrethroids were introduced in indoor residual spraying with needed success but subsequently resulted in the development of widespread multiple insecticide resistance in vectors. Vector control in many countries still use insecticides in the absence of viable alternatives. Few developments for vector control, using ovitraps, space spray, biological control agents, etc., were encouraging when used in limited scale. Likewise, recent introduction of safer vector control agents, such as insect growth regulators, biocontrol agents, and natural plant products have yet to gain the needed scale of utility for vector control. Bacterial pesticides are promising and are effective in many countries. Environmental management has shown sufficient promise for vector control and disease management but still needs advocacy for inter-sectoral coordination and sometimes are very work-intensive. The more recent genetic manipulation and sterile insect techniques are under development and consideration for use in routine vector control and for these, standardized procedures and methods are available but need thorough understanding of biology, ethical considerations, and sufficiently trained manpower for implementation being technically intensive methods. All the methods mentioned in the review that are being implemented or proposed for implementation needs effective inter-sectoral coordination and community participation. The latest strategy is evolution-proof insecticides that include fungal biopesticides, Wolbachia, and Denso virus that essentially manipulate the life cycle of the mosquitoes were found effective but needs more research. However, for effective vector control, integrated vector management methods, involving use of combination of effective tools, is needed and is also suggested by Global Malaria Control Strategy. This review article raises issues associated with the present-day vector control strategies and state opportunities with a focus on ongoing research and recent advances to enable to sustain the gains achieved so far.}, } @article {pmid21225190, year = {2010}, author = {Popovici, J and Moreira, LA and Poinsignon, A and Iturbe-Ormaetxe, I and McNaughton, D and O'Neill, SL}, title = {Assessing key safety concerns of a Wolbachia-based strategy to control dengue transmission by Aedes mosquitoes.}, journal = {Memorias do Instituto Oswaldo Cruz}, volume = {105}, number = {8}, pages = {957-964}, doi = {10.1590/s0074-02762010000800002}, pmid = {21225190}, issn = {1678-8060}, mesh = {Aedes/*microbiology ; Animals ; Dengue/prevention & control/transmission ; Dengue Virus/physiology ; Host-Parasite Interactions/*physiology ; Humans ; Insect Vectors/*microbiology ; Pest Control, Biological/*methods ; Symbiosis/physiology ; Wolbachia/*physiology ; }, abstract = {Mosquito-borne diseases such as dengue fever, chikungunya or malaria affect millions of people each year and control solutions are urgently needed. An international research program is currently being developed that relies on the introduction of the bacterial endosymbiont Wolbachia pipientis into Aedes aegypti to control dengue transmission. In order to prepare for open-field testing releases of Wolbachia-infected mosquitoes, an intensive social research and community engagement program was undertaken in Cairns, Northern Australia. The most common concern expressed by the diverse range of community members and stakeholders surveyed was the necessity of assuring the safety of the proposed approach for humans, animals and the environment. To address these concerns a series of safety experiments were undertaken. We report in this paper on the experimental data obtained, discuss the limitations of experimental risk assessment and focus on the necessity of including community concerns in scientific research.}, } @article {pmid21221516, year = {2011}, author = {Kraaijeveld, K and Reumer, BM and Mouton, L and Kremer, N and Vavre, F and van Alphen, JJ}, title = {Does a parthenogenesis-inducing Wolbachia induce vestigial cytoplasmic incompatibility?.}, journal = {Die Naturwissenschaften}, volume = {98}, number = {3}, pages = {175-180}, pmid = {21221516}, issn = {1432-1904}, mesh = {Animals ; Breeding ; Cytoplasm/*physiology ; Female ; Genes, Bacterial/genetics ; Male ; Molecular Sequence Data ; Sex Ratio ; Wasps/*microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is a maternally inherited bacterium that manipulates the reproduction of its host. Recent studies have shown that male-killing strains can induce cytoplasmic incompatibility (CI) when introgressed into a resistant host. Phylogenetic studies suggest that transitions between CI and other Wolbachia phenotypes have also occurred frequently, raising the possibility that latent CI may be widespread among Wolbachia. Here, we investigate whether a parthenogenesis-inducing Wolbachia strain can also induce CI. Parthenogenetic females of the parasitoid wasp Asobara japonica regularly produce a small number of males that may be either infected or not. Uninfected males were further obtained through removal of the Wolbachia using antibiotics and from a naturally uninfected strain. Uninfected females that had mated with infected males produced a slightly, but significantly more male-biased sex ratio than uninfected females that had mated with uninfected males. This effect was strongest in females that mated with males that had a relatively high Wolbachia titer. Quantitative PCR indicated that infected males did not show higher ratios of nuclear versus mitochondrial DNA content. Wolbachia therefore does not cause diploidization of cells in infected males. While these results are consistent with CI, other alternatives such as production of abnormal sperm by infected males cannot be completely ruled out. Overall, the effect was very small (9%), suggesting that if CI is involved it may have degenerated through the accumulation of mutations.}, } @article {pmid21214605, year = {2011}, author = {Venard, CM and Crain, PR and Dobson, SL}, title = {SYTO11 staining vs FISH staining: a comparison of two methods to stain Wolbachia pipientis in cell cultures.}, journal = {Letters in applied microbiology}, volume = {52}, number = {2}, pages = {168-176}, pmid = {21214605}, issn = {1472-765X}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; R01-AI067434/AI/NIAID NIH HHS/United States ; R01 AI067434-01A1/AI/NIAID NIH HHS/United States ; R01-AI051533/AI/NIAID NIH HHS/United States ; R01 AI051533-01A1/AI/NIAID NIH HHS/United States ; R01 AI067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/cytology ; Animals ; Cell Line ; *Fluorescent Dyes/toxicity ; *In Situ Hybridization, Fluorescence ; Wolbachia/*isolation & purification ; }, abstract = {AIMS: The Aedes albopictus C7-10 cell line was infected with Wolbachia strains wRi and wAlbB to create C7-10R and C7-10B cell lines, respectively. We compared two different methods, fluorescence in situ hybridization staining and SYTO11 staining, to describe these new Wolbachia infections in C7-10.

METHODS AND RESULTS: Both staining methods were as efficient to stain Wolbachia. A formula was developed to quantify Wolbachia infection. The infection levels in C7-10B and C7-10R differed. The live stain SYTO11 was found to be useful to visualize Wolbachia in replicating host cells. Its potential cytotoxic effect at high concentration was investigated.

CONCLUSIONS: C7-10 supported two Wolbachia infections, constituting new tools to study Wolbachia-host interactions. The different infection levels suggest that wRi and wAlbB have different requirements for their survival in C7-10 host cell line. Observation of SYTO11-stained live cells gave new insights on Wolbachia segregation pattern during host cell mitosis.

Wolbachia-induced phenotypes in their arthropod and worm hosts could potentially be used to control pest populations. However, the mechanisms underlying these phenotypes are difficult to study because of Wolbachia's intracellular lifestyle. The Wolbachia infections in C7-10 described here could be used as in vitro models to investigate Wolbachia biology.}, } @article {pmid21211038, year = {2011}, author = {Andreotti, R and Pérez de León, AA and Dowd, SE and Guerrero, FD and Bendele, KG and Scoles, GA}, title = {Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing.}, journal = {BMC microbiology}, volume = {11}, number = {1}, pages = {6}, pmid = {21211038}, issn = {1471-2180}, mesh = {Animals ; Cattle ; Corynebacterium glutamicum/classification/genetics ; Female ; Male ; Rhipicephalus/*microbiology ; Sequence Analysis, DNA ; Serratia marcescens/classification/genetics ; Staphylococcus/classification/genetics ; Staphylococcus aureus/classification/genetics ; Streptococcus/classification/genetics ; }, abstract = {BACKGROUND: Ticks are regarded as the most relevant vectors of disease-causing pathogens in domestic and wild animals. The cattle tick, Rhipicephalus (Boophilus) microplus, hinders livestock production in tropical and subtropical parts of the world where it is endemic. Tick microbiomes remain largely unexplored. The objective of this study was to explore the R. microplus microbiome by applying the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) technique to characterize its bacterial diversity. Pyrosequencing was performed on adult males and females, eggs, and gut and ovary tissues from adult females derived from samples of R. microplus collected during outbreaks in southern Texas.

RESULTS: Raw data from bTEFAP were screened and trimmed based upon quality scores and binned into individual sample collections. Bacteria identified to the species level include Staphylococcus aureus, Staphylococcus chromogenes, Streptococcus dysgalactiae, Staphylococcus sciuri, Serratia marcescens, Corynebacterium glutamicum, and Finegoldia magna. One hundred twenty-one bacterial genera were detected in all the life stages and tissues sampled. The total number of genera identified by tick sample comprised: 53 in adult males, 61 in adult females, 11 in gut tissue, 7 in ovarian tissue, and 54 in the eggs. Notable genera detected in the cattle tick include Wolbachia, Coxiella, and Borrelia. The molecular approach applied in this study allowed us to assess the relative abundance of the microbiota associated with R. microplus.

CONCLUSIONS: This report represents the first survey of the bacteriome in the cattle tick using non-culture based molecular approaches. Comparisons of our results with previous bacterial surveys provide an indication of geographic variation in the assemblages of bacteria associated with R. microplus. Additional reports on the identification of new bacterial species maintained in nature by R. microplus that may be pathogenic to its vertebrate hosts are expected as our understanding of its microbiota expands. Increased awareness of the role R. microplus can play in the transmission of pathogenic bacteria will enhance our ability to mitigate its economic impact on animal agriculture globally. This recognition should be included as part of analyses to assess the risk for re-invasion of areas like the United States of America where R. microplus was eradicated.}, } @article {pmid21203841, year = {2011}, author = {Lindh, JM and Lehane, MJ}, title = {The tsetse fly Glossina fuscipes fuscipes (Diptera: Glossina) harbours a surprising diversity of bacteria other than symbionts.}, journal = {Antonie van Leeuwenhoek}, volume = {99}, number = {3}, pages = {711-720}, doi = {10.1007/s10482-010-9546-x}, pmid = {21203841}, issn = {1572-9699}, mesh = {Animals ; Bacillus subtilis/classification/genetics ; Bacteria/classification/*genetics ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; Serratia marcescens/classification/genetics ; Symbiosis ; Tsetse Flies/*microbiology ; Wigglesworthia/classification/genetics ; }, abstract = {Three different bacterial species are regularly described from tsetse flies. However, no broad screens have been performed to investigate the existence of other bacteria in this medically and agriculturally important vector insect. Utilising both culture dependent and independent methods we show that Kenyan populations of Glossina fuscipes fuscipes harbour a surprising diversity of bacteria. Bacteria were isolated from 72% of flies with 23 different bacterial species identified. The Firmicutes phylum dominated with 16 species of which seven belong to the genus Bacillus. The tsetse fly primary symbiont, Wigglesworthia glossinidia, was identified by the culture independent pathway. However, neither the secondary symbiont Sodalis nor Wolbachia was detected with either of the methods used. Two other bacterial species were identified with the DNA based method, Bacillus subtilis and Serratia marcescens. Further studies are needed to determine how tsetse flies, which only ever feed on vertebrate blood, pick up bacteria and to investigate the possible impact of these bacteria on Glossina longevity and vector competence.}, } @article {pmid21199036, year = {2011}, author = {Toju, H and Fukatsu, T}, title = {Diversity and infection prevalence of endosymbionts in natural populations of the chestnut weevil: relevance of local climate and host plants.}, journal = {Molecular ecology}, volume = {20}, number = {4}, pages = {853-868}, doi = {10.1111/j.1365-294X.2010.04980.x}, pmid = {21199036}, issn = {1365-294X}, mesh = {Adaptation, Biological ; Animals ; *Climate ; DNA, Bacterial/genetics ; Geography ; Hippocastanaceae ; Japan ; Phylogeny ; Quercus ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*classification/physiology ; Sequence Analysis, DNA ; Spiroplasma/*classification/physiology ; *Symbiosis ; Weevils/*microbiology ; Wolbachia/*classification/physiology ; }, abstract = {Many insects are ubiquitously associated with multiple endosymbionts, whose infection patterns often exhibit spatial and temporal variations. How such endosymbiont variations are relevant to local adaptation of the host organisms is of ecological interest. Here, we report a comprehensive survey of endosymbionts in natural populations of the chestnut weevil Curculio sikkimensis, whose larvae are notorious pests of cultivated chestnuts and also infest acorns of various wild oaks. From 968 insects representing 55 localities across the Japanese Archipelago and originating from 10 host plant species, we identified six distinct endosymbiont lineages, namely Curculioniphilus, Sodalis, Serratia, Wolbachia, Rickettsia and Spiroplasma, at different infection frequencies (96.7%, 12.8%, 82.3%, 82.5%, 28.2% and 6.8%, respectively) and with different geographical distribution patterns. Multiple endosymbiont infections were very common; 3.18±0.61 (ranging from 1.74 to 5.50) endosymbionts per insect on average in each of the local populations. Five pairs of endosymbionts (Curculioniphilus-Serratia, Curculioniphilus-Wolbachia, Sodalis-Rickettsia, Wolbachia-Rickettsia and Rickettsia-Spiroplasma) co-infected the same host individuals more frequently than expected, while infections with Serratia and Wolbachia were negatively correlated to each other. Infection frequencies of the endosymbionts were significantly correlated with climatic and ecological factors: for example, higher Sodalis, Wolbachia and Rickettsia infections at localities of higher temperature; lower Wolbachia and Rickettsia infections at localities of greater snowfall; and higher Curculioniphilus, Sodalis, Serratia, Wolbachia and Rickettsia infections on acorns than on chestnuts. These patterns are discussed in relation to potential host-endosymbiont co-evolution via local adaptation across geographical populations.}, } @article {pmid21199022, year = {2011}, author = {Yu, MZ and Zhang, KJ and Xue, XF and Hong, XY}, title = {Effects of Wolbachia on mtDNA variation and evolution in natural populations of Tetranychus urticae Koch.}, journal = {Insect molecular biology}, volume = {20}, number = {3}, pages = {311-321}, doi = {10.1111/j.1365-2583.2010.01066.x}, pmid = {21199022}, issn = {1365-2583}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Electron Transport Complex IV/genetics ; Evolution, Molecular ; Haplotypes ; Phylogeny ; Polymorphism, Genetic ; Population/genetics ; Symbiosis/*genetics ; Tetranychidae/classification/*genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {We investigated the effects of Wolbachia infection on mtDNA variation in spider mites by sequencing a portion of the mitochondrial cytochrome oxidase I (COI) gene from 198 individuals of known infection status. Four Wolbachia strains were described in the current study, namely wUrtOri1, wUrtOri2, wUrtOri3 and wUrtCon1. As predicted, the haplotype and nucleotide diversity were lower in infected individuals than in uninfected individuals. However, these mtDNA haplotype data are not entirely concordant with the surface protein of wolbachia (wsp) sequence data and both infected and uninfected individuals were found of the same haplotype. Although values of Tajima's D and Fu & Li's F were consistently less than zero for most infected groups, McDonald-Kreitman tests suggested that the patterns of variation were different from those expected under neutrality in only the uninfected group. Thus, the neutrality tests do not show a clear effect of Wolbachia infection on patterns of mtDNA variation and substitution in spider mites.}, } @article {pmid26467727, year = {2011}, author = {Jenkins, TM and Eaton, TD}, title = {Population Genetic Baseline of the First Plataspid Stink Bug Symbiosis (Hemiptera: Heteroptera: Plataspidae) Reported in North America.}, journal = {Insects}, volume = {2}, number = {3}, pages = {264-272}, pmid = {26467727}, issn = {2075-4450}, abstract = {The stink bug, Megacopta cribraria, has an obligate relationship with a bacterial endosymbiont which allows it to feed on legumes. The insect is a pest of soybeans in Asia and was first reported in the Western Hemisphere in October 2009 on kudzu vine, Pueraria montana, in North Georgia, USA. By October 2010 M. cribraria had been confirmed in 80 counties in Georgia actively feeding on kudzu vine and soybean plants. Since the symbiosis may support the bug's ecological expansions, a population genetic baseline for the symbiosis was developed from mitochondrial DNA (mtDNA) and nuclear DNA (nuDNA) gene sequence collected from each insect and its primary g- proteobacterium and secondary a -proteobacterium endosymbionts. A single mitochondrial DNA haplotype was found in all insects sampled in Georgia and South Carolina identified as GA1. The GAI haplotype appears to be rapidly dispersing across Georgia and into contiguous states. Primary and secondary endosymbiont gene sequences from M. cribraria in Georgia were the same as those found in recently collected Megacopta samples from Japan. The implications of these data are discussed.}, } @article {pmid22654845, year = {2011}, author = {Negri, I}, title = {Wolbachia as an "infectious" extrinsic factor manipulating host signaling pathways.}, journal = {Frontiers in endocrinology}, volume = {2}, number = {}, pages = {115}, pmid = {22654845}, issn = {1664-2392}, abstract = {Wolbachia pipientis is a widespread endosymbiont of filarial nematodes and arthropods. While in worms the symbiosis is obligate, in arthropods Wolbachia induces several reproductive manipulations (i.e., cytoplasmic incompatibility, parthenogenesis, feminization of genetic males, and male-killing) in order to increase the number of infected females. These various phenotypic effects may be linked to differences in host physiology, and in particular to endocrine-related processes governing growth, development, and reproduction. Indeed, a number of evidences links Wolbachia symbiosis to insulin and ecdysteroid signaling, two multilayered pathways known to work antagonistically, jointly or even independently for the regulation of different molecular networks. At present it is not clear whether Wolbachia manipulates one pathway, thus affecting other related metabolic networks, or if it targets both pathways, even interacting at several points in each of them. Interestingly, in view of the interplay between hormone signaling and epigenetic machinery, a direct influence of the "infection" on hormonal signaling involving ecdysteroids might be achievable through the manipulation of the host's epigenetic pathways.}, } @article {pmid21187977, year = {2010}, author = {Ling, A and Cordaux, R}, title = {Insertion sequence inversions mediated by ectopic recombination between terminal inverted repeats.}, journal = {PloS one}, volume = {5}, number = {12}, pages = {e15654}, pmid = {21187977}, issn = {1932-6203}, mesh = {Base Sequence ; Chromosome Inversion ; DNA/genetics ; DNA Transposable Elements ; Genome ; Genomic Instability ; Molecular Sequence Data ; Phylogeny ; Promoter Regions, Genetic ; *Recombination, Genetic ; Sequence Homology, Nucleic Acid ; *Sequence Inversion ; *Terminal Repeat Sequences ; Wolbachia/genetics ; }, abstract = {Transposable elements are widely distributed and diverse in both eukaryotes and prokaryotes, as exemplified by DNA transposons. As a result, they represent a considerable source of genomic variation, for example through ectopic (i.e. non-allelic homologous) recombination events between transposable element copies, resulting in genomic rearrangements. Ectopic recombination may also take place between homologous sequences located within transposable element sequences. DNA transposons are typically bounded by terminal inverted repeats (TIRs). Ectopic recombination between TIRs is expected to result in DNA transposon inversions. However, such inversions have barely been documented. In this study, we report natural inversions of the most common prokaryotic DNA transposons: insertion sequences (IS). We identified natural TIR-TIR recombination-mediated inversions in 9% of IS insertion loci investigated in Wolbachia bacteria, which suggests that recombination between IS TIRs may be a quite common, albeit largely overlooked, source of genomic diversity in bacteria. We suggest that inversions may impede IS survival and proliferation in the host genome by altering transpositional activity. They may also alter genomic instability by modulating the outcome of ectopic recombination events between IS copies in various orientations. This study represents the first report of TIR-TIR recombination within bacterial IS elements and it thereby uncovers a novel mechanism of structural variation for this class of prokaryotic transposable elements.}, } @article {pmid21183640, year = {2011}, author = {Gonella, E and Negri, I and Marzorati, M and Mandrioli, M and Sacchi, L and Pajoro, M and Crotti, E and Rizzi, A and Clementi, E and Tedeschi, R and Bandi, C and Alma, A and Daffonchio, D}, title = {Bacterial endosymbiont localization in Hyalesthes obsoletus, the insect vector of Bois noir in Vitis vinifera.}, journal = {Applied and environmental microbiology}, volume = {77}, number = {4}, pages = {1423-1435}, pmid = {21183640}, issn = {1098-5336}, mesh = {Animals ; Bacteroidetes/classification/isolation & purification ; Base Sequence ; Betaproteobacteria/classification/isolation & purification ; Denaturing Gradient Gel Electrophoresis ; Gammaproteobacteria/classification/isolation & purification ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Insect Vectors/*microbiology ; Italy ; Microbial Consortia ; Molecular Sequence Data ; Phytoplasma/*pathogenicity ; Plant Diseases/*microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsia/classification/isolation & purification ; Sequence Analysis, DNA ; *Symbiosis ; Vitis/*microbiology ; }, abstract = {One emerging disease of grapevine in Europe is Bois noir (BN), a phytoplasmosis caused by "Candidatus Phytoplasma solani" and spread in vineyards by the planthopper Hyalesthes obsoletus (Hemiptera: Cixiidae). Here we present the first full characterization of the bacterial community of this important disease vector collected from BN-contaminated areas in Piedmont, Italy. Length heterogeneity PCR and denaturing gradient gel electrophoresis analysis targeting the 16S rRNA gene revealed the presence of a number of bacteria stably associated with the insect vector. In particular, symbiotic bacteria detected by PCR with high infection rates in adult individuals fell within the "Candidatus Sulcia muelleri" cluster in the Bacteroidetes and in the "Candidatus Purcelliella pentastirinorum" group in the Gammaproteobacteria, both previously identified in different leafhoppers and planthoppers. A high infection rate (81%) was also shown for another symbiont belonging to the Betaproteobacteria, designated the HO1-V symbiont. Because of the low level of 16S rRNA gene identity (80%) with the closest relative, an uncharacterized symbiont of the tick Haemaphysalis longicornis, we propose the new name "Candidatus Vidania fulgoroideae." Other bacterial endosymbionts identified in H. obsoletus were related to the intracellular bacteria Wolbachia pipientis, Rickettsia sp., and "Candidatus Cardinium hertigii." Fluorescent in situ hybridization coupled with confocal laser scanning microscopy and transmission electron microscopy showed that these bacteria are localized in the gut, testicles, and oocytes. As "Ca. Sulcia" is usually reported in association with other symbiotic bacteria, we propose that in H. obsoletus, it may occur in a bipartite or even tripartite relationship between "Ca. Sulcia" and "Ca. Purcelliella," "Ca. Vidania," or both.}, } @article {pmid21177682, year = {2011}, author = {Hansen, RD and Trees, AJ and Bah, GS and Hetzel, U and Martin, C and Bain, O and Tanya, VN and Makepeace, BL}, title = {A worm's best friend: recruitment of neutrophils by Wolbachia confounds eosinophil degranulation against the filarial nematode Onchocerca ochengi.}, journal = {Proceedings. Biological sciences}, volume = {278}, number = {1716}, pages = {2293-2302}, pmid = {21177682}, issn = {1471-2954}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Arsenicals/*pharmacology/therapeutic use ; Cameroon ; Cattle ; Cattle Diseases/*drug therapy/*parasitology ; Cell Degranulation/drug effects/*physiology ; Eosinophils/physiology ; Gene Expression Regulation/drug effects ; Histological Techniques ; Neutrophils/drug effects/*physiology ; Onchocerca/*microbiology ; Onchocerciasis/drug therapy/*veterinary ; Oxytetracycline/*pharmacology/therapeutic use ; Triazines/*pharmacology/therapeutic use ; Wolbachia/drug effects/immunology/*physiology ; }, abstract = {Onchocerca ochengi, a filarial parasite of cattle, represents the closest relative of the human pathogen, Onchocerca volvulus. Both species harbour Wolbachia endosymbionts and are remarkable in that adult female worms remain viable but sessile for many years while surrounded by host cells and antibodies. The basis of the symbiosis between filariae and Wolbachia is thought to be metabolic, although a role for Wolbachia in immune evasion has received little attention. Neutrophils are attracted to Wolbachia, but following antibiotic chemotherapy they are replaced by eosinophils that degranulate on the worm cuticle. However, it is unclear whether the eosinophils are involved in parasite killing or if they are attracted secondarily to dying worms. In this study, cattle infected with Onchocerca ochengi received adulticidal regimens of oxytetracycline or melarsomine. In contrast to oxytetracycline, melarsomine did not directly affect Wolbachia viability. Eosinophil degranulation increased significantly only in the oxytetracycline group; whereas nodular gene expression of bovine neutrophilic chemokines was lowest in this group. Moreover, intense eosinophil degranulation was initially associated with worm vitality, not degeneration. Taken together, these data offer strong support for the hypothesis that Wolbachia confers longevity on O. ochengi through a defensive mutualism, which diverts a potentially lethal effector cell response.}, } @article {pmid21175905, year = {2011}, author = {Martinson, VG and Danforth, BN and Minckley, RL and Rueppell, O and Tingek, S and Moran, NA}, title = {A simple and distinctive microbiota associated with honey bees and bumble bees.}, journal = {Molecular ecology}, volume = {20}, number = {3}, pages = {619-628}, doi = {10.1111/j.1365-294X.2010.04959.x}, pmid = {21175905}, issn = {1365-294X}, mesh = {Animals ; Bacteria/classification/*genetics ; Base Sequence ; Bees/*microbiology ; Burkholderia/genetics ; Cloning, Molecular ; DNA, Bacterial/genetics/isolation & purification ; DNA, Ribosomal/genetics ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial ; RNA, Ribosomal, 16S/*genetics ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Wasps/*microbiology ; Wolbachia/genetics ; }, abstract = {Specialized relationships with bacteria often allow animals to exploit a new diet by providing a novel set of metabolic capabilities. Bees are a monophyletic group of Hymenoptera that transitioned to a completely herbivorous diet from the carnivorous diet of their wasp ancestors. Recent culture-independent studies suggest that a set of distinctive bacterial species inhabits the gut of the honey bee, Apis mellifera. Here we survey the gut microbiotae of diverse bee and wasp species to test whether acquisition of these bacteria was associated with the transition to herbivory in bees generally. We found that most bee species lack phylotypes that are the same or similar to those typical of A. mellifera, rejecting the hypothesis that this dietary transition was symbiont-dependent. The most common bacteria in solitary bee species are a widespread phylotype of Burkholderia and the pervasive insect associate, Wolbachia. In contrast, several social representatives of corbiculate bees do possess distinctive bacterial phylotypes. Samples of A. mellifera harboured the same microbiota as in previous surveys, and closely related bacterial phylotypes were identified in two Asian honey bees (Apis andreniformis and Apis dorsata) and several bumble bee (Bombus) species. Potentially, the sociality of Apis and Bombus species facilitates symbiont transmission and thus is key to the maintenance of a more consistent gut microbiota. Phylogenetic analyses provide a more refined taxonomic placement of the A. mellifera symbionts.}, } @article {pmid21175696, year = {2011}, author = {Zouache, K and Raharimalala, FN and Raquin, V and Tran-Van, V and Raveloson, LH and Ravelonandro, P and Mavingui, P}, title = {Bacterial diversity of field-caught mosquitoes, Aedes albopictus and Aedes aegypti, from different geographic regions of Madagascar.}, journal = {FEMS microbiology ecology}, volume = {75}, number = {3}, pages = {377-389}, doi = {10.1111/j.1574-6941.2010.01012.x}, pmid = {21175696}, issn = {1574-6941}, mesh = {Aedes/*microbiology ; Animals ; Bacteria/*classification/genetics/growth & development/isolation & purification ; *Biodiversity ; Denaturing Gradient Gel Electrophoresis ; *Ecosystem ; Female ; Madagascar ; Male ; Molecular Sequence Data ; Phylogeny ; Principal Component Analysis ; RNA, Ribosomal, 16S/genetics ; Wolbachia/classification/genetics/physiology ; }, abstract = {Symbiotic bacteria are known to play important roles in the biology of insects, but the current knowledge of bacterial communities associated with mosquitoes is very limited and consequently their contribution to host behaviors is mostly unknown. In this study, we explored the composition and diversity of mosquito-associated bacteria in relation with mosquitoes' habitats. Wild Aedes albopictus and Aedes aegypti were collected in three different geographic regions of Madagascar. Culturing methods and denaturing gradient gel electrophoresis (DGGE) and sequencing of the rrs amplicons revealed that Proteobacteria and Firmicutes were the major phyla. Isolated bacterial genera were dominated by Bacillus, followed by Acinetobacter, Agrobacterium and Enterobacter. Common DGGE bands belonged to Acinetobacter, Asaia, Delftia, Pseudomonas, Enterobacteriaceae and an uncultured Gammaproteobacterium. Double infection by maternally inherited Wolbachia pipientis prevailed in 98% of males (n=272) and 99% of females (n=413); few individuals were found to be monoinfected with Wolbachia wAlbB strain. Bacterial diversity (Shannon-Weaver and Simpson indices) differed significantly per habitat whereas evenness (Pielou index) was similar. Overall, the bacterial composition and diversity were influenced both by the sex of individuals and by the environment inhabited by the mosquitoes; the latter might be related to both the vegetation and the animal host populations that Aedes used as food sources.}, } @article {pmid21175057, year = {2010}, author = {Bellini, R and Albieri, A and Balestrino, F and Carrieri, M and Porretta, D and Urbanelli, S and Calvitti, M and Moretti, R and Maini, S}, title = {Dispersal and survival of Aedes albopictus (Diptera: Culicidae) males in Italian urban areas and significance for sterile insect technique application.}, journal = {Journal of medical entomology}, volume = {47}, number = {6}, pages = {1082-1091}, doi = {10.1603/me09154}, pmid = {21175057}, issn = {0022-2585}, mesh = {Aedes/*physiology ; Animals ; Demography ; Italy ; Longevity ; Male ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; Urbanization ; }, abstract = {The dispersal and survival of laboratory-reared Aedes albopictus Skuse males were investigated during the summer of 2007 in three Northern Italy urban localities by mark-release-recapture techniques. Two marking methods were compared: one group of males was dusted with fluorescent pigments on the body (FP), and the other group was obtained from a strain whose natural infection of Wolbachia had been removed (WB0). FP- and WB0-marked males were released as adults and pupae, respectively, in one fixed station at each locality. Recaptures were performed by skilled technicians, within a radius of 350 m from the release site, on days 4, 5, and 7 after the release, and the males were collected while flying around the technician's body or in swarms. Recapture rates ranged from 0.63 to 4.72% for FP males and from 2.39 to 11.05% for WB0 males. The mean distance traveled for WB0 males was significantly higher than for FP males; no difference was observed between the dispersal distance measured for the males recaptured on human host versus males recaptured while swarming. No further increase of the dispersal occurred during the postrelease period investigated (from day 4 to day 7 after release). The mean survival rate at the release was 0.51 for FP-marked males and 0.81 for WB0 males. The data obtained are discussed for their significance in planning sterile insect technique programs against Ae. albopictus.}, } @article {pmid21174352, year = {2010}, author = {Hafer, N and Pike, N}, title = {Shape change in viable eggs of the collembolan Folsomia candida provides insight into the role of Wolbachia endosymbionts.}, journal = {Dong wu xue yan jiu = Zoological research}, volume = {31}, number = {6}, pages = {623-626}, doi = {10.3724/SP.J.1141.2010.06623}, pmid = {21174352}, issn = {0254-5853}, mesh = {Animals ; Arthropods/*microbiology ; Female ; Ovum/cytology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacteria of the genus Wolbachia that infect the collembolan species Folsomia candida are responsible for facilitating parthenogenetic reproduction in their hosts. This study made empirical observations of the development of eggs of F. candida which contained normal populations of Wolbachia and of eggs which were cured of Wolbachia by treatment with the antibiotic rifampicin. A marked increase in egg size accompanied by a significant change in shape from spherical to discoid occurred in viable eggs three to four days after laying. These changes did not occur in the universally inviable eggs which came from the antibiotic treatment or in the 7% of untreated eggs which were naturally inviable. We infer that Wolbachia plays a critical role in zygotic or embryonic development during or before the first three days after laying and we draw on existing knowledge in speculating on the developmental mechanisms that Wolbachia may influence.}, } @article {pmid21171866, year = {2010}, author = {Belanger, DH and Perkins, SL}, title = {Wolbachia infection and mitochondrial diversity in the canine heartworm (Dirofilaria immitis).}, journal = {Mitochondrial DNA}, volume = {21}, number = {6}, pages = {227-233}, doi = {10.3109/19401736.2010.533765}, pmid = {21171866}, issn = {1940-1744}, mesh = {Animals ; Carnivora/parasitology ; DNA, Helminth/genetics ; DNA, Mitochondrial/*genetics ; Dirofilaria immitis/cytology/*genetics/*microbiology ; *Genetic Variation ; North America ; Wolbachia/*physiology ; }, abstract = {BACKGROUND AND AIMS: Many species of filarial nematodes are infected with Wolbachia pipientis, a maternally inherited endosymbiont. In addition to manipulating host reproduction, these bacteria also affect the evolution of the mitochondrial DNA with which they are transmitted. Selective sweeps can establish a single mitochondrial lineage within a Wolbachia-infected population and purge genetic diversity. While this phenomenon has been studied in insect model systems, it has not been thoroughly examined in a filarial nematode.

MATERIALS AND METHODS: Patterns of mitochondrial diversity were examined in Dirofilaria immitis, a Wolbachia-infected species.

RESULTS: The levels of genetic diversity observed in canine heartworm were much lower than those in related species not known to be hosts of Wolbachia.

CONCLUSION: RESULTS suggest that a maternally inherited endosymbiont can depress mitochondrial diversity in a filarial host.}, } @article {pmid21151959, year = {2010}, author = {Miller, WJ and Ehrman, L and Schneider, D}, title = {Infectious speciation revisited: impact of symbiont-depletion on female fitness and mating behavior of Drosophila paulistorum.}, journal = {PLoS pathogens}, volume = {6}, number = {12}, pages = {e1001214}, pmid = {21151959}, issn = {1553-7374}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Biological Evolution ; Drosophila/*microbiology ; Female ; *Fertility ; *Sexual Behavior, Animal ; *Symbiosis ; Wolbachia ; }, abstract = {The neotropical Drosophila paulistorum superspecies, consisting of at least six geographically overlapping but reproductively isolated semispecies, has been the object of extensive research since at least 1955, when it was initially trapped mid-evolution in flagrant statu nascendi. In this classic system females express strong premating isolation patterns against mates belonging to any other semispecies, and yet uncharacterized microbial reproductive tract symbionts were described triggering hybrid inviability and male sterility. Based on theoretical models and limited experimental data, prime candidates fostering symbiont-driven speciation in arthropods are intracellular bacteria belonging to the genus Wolbachia. They are maternally inherited symbionts of many arthropods capable of manipulating host reproductive biology for their own benefits. However, it is an ongoing debate as to whether or not reproductive symbionts are capable of driving host speciation in nature and if so, to what extent. Here we have reevaluated this classic case of infectious speciation by means of present day molecular approaches and artificial symbiont depletion experiments. We have isolated the α-proteobacteria Wolbachia as the maternally transmitted core endosymbionts of all D. paulistorum semispecies that have coevolved towards obligate mutualism with their respective native hosts. In hybrids, however, these mutualists transform into pathogens by overreplication causing embryonic inviability and male sterility. We show that experimental reduction in native Wolbachia titer causes alterations in sex ratio, fecundity, and mate discrimination. Our results indicate that formerly designated Mycoplasma-like organisms are most likely Wolbachia that have evolved by becoming essential mutualistic symbionts in their respective natural hosts; they have the potential to trigger pre- and postmating isolation. Furthermore, in light of our new findings, we revisit the concept of infectious speciation and discuss potential mechanisms that can restrict or promote symbiont-induced speciation at post- and prezygotic levels in nature and under artificial laboratory conditions.}, } @article {pmid21148356, year = {2010}, author = {Enserink, M}, title = {Infectious diseases. Australia to test 'mosquito vaccine' against human disease.}, journal = {Science (New York, N.Y.)}, volume = {330}, number = {6010}, pages = {1460-1461}, doi = {10.1126/science.330.6010.1460}, pmid = {21148356}, issn = {1095-9203}, mesh = {Aedes/*microbiology/*virology ; Animals ; Dengue/prevention & control/*transmission ; Dengue Virus/*physiology ; Female ; Humans ; Insect Vectors/*microbiology/virology ; Male ; Queensland ; Wolbachia/*physiology ; }, } @article {pmid21147476, year = {2010}, author = {Lee, AC and Atkins, CE}, title = {Understanding feline heartworm infection: disease, diagnosis, and treatment.}, journal = {Topics in companion animal medicine}, volume = {25}, number = {4}, pages = {224-230}, doi = {10.1053/j.tcam.2010.09.003}, pmid = {21147476}, issn = {1946-9837}, mesh = {Animals ; Cat Diseases/*diagnosis/diagnostic imaging/surgery ; Cats ; Dirofilaria immitis/pathogenicity/*physiology ; Dirofilariasis/*diagnosis/diagnostic imaging/surgery ; Echocardiography/veterinary ; Female ; Life Cycle Stages ; Male ; Prognosis ; Pulmonary Artery/diagnostic imaging/pathology ; Radiography, Thoracic/veterinary ; Remission, Spontaneous ; Risk Factors ; }, abstract = {Feline heartworm disease is a very different clinical entity from canine heartworm disease. In cats, the arrival and death of immature heartworms in the pulmonary arteries can cause coughing and dyspnea as early as 3 months postinfection. Adult heartworms suppress the function of pulmonary intravascular macrophages and thus reduce clinical disease in chronic feline heartworm infection. Approximately 80% of asymptomatic cats self-cure. Median survival time for symptomatic cats is 1.5 years, or 4 years if only cats living beyond the day of presentation are considered. Aberrant worm migration is more frequent than it is in dogs, and sudden death can occur with no prior clinical signs. The bacterial endosymbiont Wolbachia likely contributes to the inflammatory pathology of heartworm disease, but its role is not yet fully clear. Unfortunately, the diagnosis, treatment, and management of feline heartworm disease are far from simple. Antemortem diagnosis is hampered by low worm burdens, the frequency of all-male infections, and nonspecific radiographic lesions. It is up to the veterinarian to determine the correct index of suspicion and choose the right combination of diagnostic tests to achieve an answer. Treatment is symptomatic because adulticide therapy is risky and does not increase survival time. Despite the dangers of feline heartworm disease, less than 5% of cats in the United States are on chemoprophylaxis. It is important for veterinarians to take a proactive preventive stance because heartworm infection in cats is a multisystemic disease that has no easy cure.}, } @article {pmid21135075, year = {2011}, author = {Yeap, HL and Mee, P and Walker, T and Weeks, AR and O'Neill, SL and Johnson, P and Ritchie, SA and Richardson, KM and Doig, C and Endersby, NM and Hoffmann, AA}, title = {Dynamics of the "popcorn" Wolbachia infection in outbred Aedes aegypti informs prospects for mosquito vector control.}, journal = {Genetics}, volume = {187}, number = {2}, pages = {583-595}, pmid = {21135075}, issn = {1943-2631}, mesh = {Aedes/genetics/*microbiology ; Animals ; Drosophila melanogaster/microbiology ; Female ; Genetic Fitness ; Genetic Variation ; *Insect Vectors/genetics/microbiology ; Longevity ; *Mosquito Control ; *Population Dynamics ; Survival Analysis ; Wolbachia/*physiology ; }, abstract = {Forty percent of the world's population is at risk of contracting dengue virus, which produces dengue fever with a potentially fatal hemorrhagic form. The wMelPop Wolbachia infection of Drosophila melanogaster reduces life span and interferes with viral transmission when introduced into the mosquito Aedes aegypti, the primary vector of dengue virus. Wolbachia has been proposed as an agent for preventing transmission of dengue virus. Population invasion by Wolbachia depends on levels of cytoplasmic incompatibility, fitness effects, and maternal transmission. Here we characterized these traits in an outbred genetic background of a potential target population of Ae. aegypti using two crossing schemes. Cytoplasmic incompatibility was strong in this background, and the maternal transmission rate of Wolbachia was high. The infection substantially reduced longevity of infected adult females, regardless of whether adults came from larvae cultured under high or low levels of nutrition or density. The infection reduced the viability of diapausing and nondiapausing eggs. Viability was particularly low when eggs were laid by older females and when diapausing eggs had been stored for a few weeks. The infection affected mosquito larval development time and adult body size under different larval nutrition levels and densities. The results were used to assess the potential for wMelPop-CLA to invade natural populations of Ae. aegypti and to develop recommendations for the maintenance of fitness in infected mosquitoes that need to compete against field insects.}, } @article {pmid21124735, year = {2010}, author = {Xiao, JH and Wang, NX and Li, YW and Murphy, RW and Wan, DG and Niu, LM and Hu, HY and Fu, YG and Huang, DW}, title = {Molecular approaches to identify cryptic species and polymorphic species within a complex community of fig wasps.}, journal = {PloS one}, volume = {5}, number = {11}, pages = {e15067}, pmid = {21124735}, issn = {1932-6203}, mesh = {Animals ; Bayes Theorem ; Cell Nucleus/genetics ; Cytochromes b/*genetics ; DNA, Mitochondrial/chemistry/genetics ; DNA, Ribosomal Spacer/*genetics ; Ecosystem ; Electron Transport Complex IV/*genetics ; Female ; Ficus/*parasitology ; Insect Proteins/genetics ; Male ; Molecular Sequence Data ; Peptide Elongation Factor 1/*genetics ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; Wasps/classification/*genetics ; }, abstract = {Cryptic and polymorphic species can complicate traditional taxonomic research and both of these concerns are common in fig wasp communities. Species identification is very difficult, despite great effort and the ecological importance of fig wasps. Herein, we try to identify all chalcidoid wasp species hosted by one species of fig, using both morphological and molecular methods. We compare the efficiency of four different DNA regions and find that ITS2 is highly effective for species identification, while mitochondrial COI and Cytb regions appear less reliable, possibly due to the interference signals from either nuclear copies of mtDNA, i.e. NUMTs, or the effects of Wolbachia infections. The analyses suggest that combining multiple markers is the best choice for inferring species identifications as any one marker may be unsuitable in a given case.}, } @article {pmid21119702, year = {2011}, author = {Duron, O and Raymond, M and Weill, M}, title = {Many compatible Wolbachia strains coexist within natural populations of Culex pipiens mosquito.}, journal = {Heredity}, volume = {106}, number = {6}, pages = {986-993}, pmid = {21119702}, issn = {1365-2540}, mesh = {Animals ; Breeding ; Culex/genetics/*microbiology ; Female ; France ; Genetic Variation/*genetics ; Genetics, Population ; *Host-Pathogen Interactions ; Male ; Reproduction/genetics ; Wolbachia/genetics/*physiology ; }, abstract = {Maternally inherited Wolbachia often manipulate the reproduction of arthropods to promote their transmission. In most species, Wolbachia exert a form of conditional sterility termed cytoplasmic incompatibility (CI), characterized by the death of embryos produced by the mating between individuals with incompatible Wolbachia infections. From a theoretical perspective, no stable coexistence of incompatible Wolbachia infections is expected within host populations and CI should induce the invasion of one strain or of a set of compatible strains. In this study, we investigated this prediction on CI dynamics in natural populations of the common house mosquito Culex pipiens. We surveyed the Wolbachia diversity and the expression of CI in breeding sites of the south of France between 1990 and 2005. We found that geographically close C. pipiens populations harbor considerable Wolbachia diversity, which is stably maintained over 15 years. We also observed a very low frequency of infertile clutches within each sampled site. Meanwhile, mating choice experiments conducted in laboratory conditions showed that assortative mating does not occur. Overall, this suggests that a large set of compatible Wolbachia strains are always locally dominant within mosquito populations thus, fitting with the theoretical expectations on CI dynamics.}, } @article {pmid21114563, year = {2011}, author = {Atyame, CM and Duron, O and Tortosa, P and Pasteur, N and Fort, P and Weill, M}, title = {Multiple Wolbachia determinants control the evolution of cytoplasmic incompatibilities in Culex pipiens mosquito populations.}, journal = {Molecular ecology}, volume = {20}, number = {2}, pages = {286-298}, doi = {10.1111/j.1365-294X.2010.04937.x}, pmid = {21114563}, issn = {1365-294X}, mesh = {Animals ; Biological Evolution ; Crosses, Genetic ; Culex/genetics/*microbiology/*physiology ; Cytoplasm/physiology ; DNA, Mitochondrial/genetics ; Female ; Genetic Markers ; Indian Ocean ; Male ; Mitosis ; Polymerase Chain Reaction ; *Polymorphism, Genetic ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia are maternally inherited endosymbionts that can invade arthropod populations through manipulation of their reproduction. In mosquitoes, Wolbachia induce embryonic death, known as cytoplasmic incompatibility (CI), whenever infected males mate with females either uninfected or infected with an incompatible strain. Although genetic determinants of CI are unknown, a functional model involving the so-called mod and resc factors has been proposed. Natural populations of Culex pipiens mosquito display a complex CI relationship pattern associated with the highest Wolbachia (wPip) genetic polymorphism reported so far. We show here that C. pipiens populations from La Réunion, a geographically isolated island in the southwest of the Indian Ocean, are infected with genetically closely related wPip strains. Crossing experiments reveal that these Wolbachia are all mutually compatible. However, crosses with genetically more distant wPip strains indicate that Wolbachia strains from La Réunion belong to at least five distinct incompatibility groups (or crossing types). These incompatibility properties which are strictly independent from the nuclear background, formally establish that in C. pipiens, CI is controlled by several Wolbachia mod/resc factors.}, } @article {pmid21114562, year = {2011}, author = {Caragata, EP and Poinsignon, A and Moreira, LA and Johnson, PH and Leong, YS and Ritchie, SA and O'Neill, SL and McGraw, EA}, title = {Improved accuracy of the transcriptional profiling method of age grading in Aedes aegypti mosquitoes under laboratory and semi-field cage conditions and in the presence of Wolbachia infection.}, journal = {Insect molecular biology}, volume = {20}, number = {2}, pages = {215-224}, doi = {10.1111/j.1365-2583.2010.01059.x}, pmid = {21114562}, issn = {1365-2583}, mesh = {Aedes/genetics/*microbiology/*physiology ; Aging/genetics ; Animals ; Australia ; Female ; Gene Expression Profiling/*methods ; Genes, Insect ; Models, Biological ; Symbiosis ; Transcription, Genetic ; Wolbachia/*physiology ; }, abstract = {Transcriptional profiling is an effective method of predicting age in the mosquito Aedes aegypti in the laboratory, however, its effectiveness is limited to younger mosquitoes. To address this we used a microarray to identify new gene candidates that show significant expression changes in older mosquitoes. These genes were then used to create a revised model, which upon evaluation in both laboratory and semi-field conditions, proved to have improved accuracy overall and for older mosquitoes. In association with the development of symbiont-based control strategies for Ae. aegypti, we also tested the model's accuracy for Wolbachia-infected mosquitoes and found no decline in performance. Our findings suggest that the new model is a robust and powerful tool for age determination in Australian Ae. aegypti populations.}, } @article {pmid21075139, year = {2011}, author = {Thomas, P and Kenny, N and Eyles, D and Moreira, LA and O'Neill, SL and Asgari, S}, title = {Infection with the wMel and wMelPop strains of Wolbachia leads to higher levels of melanization in the hemolymph of Drosophila melanogaster, Drosophila simulans and Aedes aegypti.}, journal = {Developmental and comparative immunology}, volume = {35}, number = {3}, pages = {360-365}, doi = {10.1016/j.dci.2010.11.007}, pmid = {21075139}, issn = {1879-0089}, mesh = {Aedes/immunology/*parasitology ; Animals ; Chromatography, High Pressure Liquid ; Dopamine/metabolism ; Drosophila/immunology/*parasitology ; Female ; Hemolymph/*metabolism ; Host-Parasite Interactions ; Male ; Melanins/*metabolism ; Wolbachia/*immunology ; }, abstract = {Introduction of the life-shortening strain of Wolbachia pipientis, wMelPop, into the key dengue vector, Aedes aegypti, and the anti-pathogen effects in Wolbachia-infected hosts highlights the need for more research into its interactions with its original host, Drosophila melanogaster, and the novel mosquito host. The visual difference in darkness between the eggs of wMelPop Wolbachia-infected and uninfected mosquito hosts after egg deposition led to further investigation into melanization levels of the insects. Both D. melanogaster and A. aegypti infected with wMelPop showed increased levels of melanization, especially in females. This result was also seen in D. melanogaster and Drosophila simulans infected with the closely related wMel strain. D. simulans infected with other strains of Wolbachia did not display this difference. HPLC analysis of hemolymph from mosquitoes showed that this difference was not due to dopamine levels in the host as they were no different in wMelPop-infected and control mosquitoes before or after blood feeding.}, } @article {pmid21074526, year = {2011}, author = {Pechgit, P and Intarapuk, A and Pinyoowong, D and Bhumiratana, A}, title = {Touchdown-touchup nested PCR for low-copy gene detection of benzimidazole-susceptible Wuchereria bancrofti with a Wolbachia endosymbiont imported by migrant carriers.}, journal = {Experimental parasitology}, volume = {127}, number = {2}, pages = {559-568}, doi = {10.1016/j.exppara.2010.10.022}, pmid = {21074526}, issn = {1090-2449}, mesh = {Adult ; Algorithms ; Animals ; Anthelmintics/pharmacology ; Benzimidazoles/pharmacology ; Carrier State/*diagnosis/ethnology/parasitology/transmission ; DNA, Bacterial/blood ; Elephantiasis, Filarial/*diagnosis/ethnology/transmission ; Female ; Humans ; Male ; Myanmar/ethnology ; Polymerase Chain Reaction ; Symbiosis ; Thailand ; *Transients and Migrants ; Wolbachia/*physiology ; Wuchereria bancrofti/drug effects/genetics/*isolation & purification/microbiology ; Young Adult ; }, abstract = {A novel, sensitive and specific touchdown-touchup nested PCR (TNPCR) technique based on two useful molecular markers, a Wuchereria bancrofti β-tubulin gene involved in benzimidazole susceptibility and a Wolbachia ftsZ gene involved in cell division, was developed to simultaneously detect the parasite W. bancrofti (W1) with its Wolbachia endosymbiont (W2) from both microfilaremic and post-treatment samples of at-risk migrant carriers infected with geographical W. bancrofti isolates. The detection and characterization of authentically low-copy gene-derived amplicons revealed no false positive identifications in amicrofilaremia with or without antigenemia. The W1-TNPCR was 100-fold more sensitive than the W2-TNPCR regardless of the microfilarial DNA isolation method and compared well with the thick blood film and membrane filtration techniques. These locus-specific TNPCRs could also detect Wolbachia-carrying W. bancrofti genotype in addition to a link to benzimidazole sensitivity among those with unknown infection origins that exhibited microfilaremia responsiveness against treatment with diethylcarbamazine plus albendazole. These TNPCR methods can augment the results of microscopic detection of the parasite because these methods enhance DNA isolation and PCR amplification capabilities.}, } @article {pmid21073054, year = {2010}, author = {Sanprasert, V and Sujariyakul, A and Nuchprayoon, S}, title = {A single dose of doxycycline in combination with diethylcarbamazine for treatment of bancroftian filariasis.}, journal = {The Southeast Asian journal of tropical medicine and public health}, volume = {41}, number = {4}, pages = {800-812}, pmid = {21073054}, issn = {0125-1562}, mesh = {Adolescent ; Adult ; Animals ; Diethylcarbamazine/*administration & dosage/adverse effects ; Double-Blind Method ; Doxycycline/*administration & dosage/adverse effects ; Drug Administration Schedule ; Drug Therapy, Combination ; Elephantiasis, Filarial/*drug therapy ; Female ; Filaricides/*administration & dosage/adverse effects ; Humans ; Male ; Middle Aged ; *Wuchereria bancrofti ; Young Adult ; }, abstract = {Standard treatment of lymphatic filariasis with diethylcarbamazine (DEC) is associated with systemic adverse reactions, thought to be due to the release of microfilariae material and Wolbachia endosymbiotic bacteria into the blood. Combination treatments with doxycycline for 3-8 weeks are more effective than standard treatment. However, long-term use of antibiotics may contribute to drug resistance and are not practical for use in remote areas. We assessed whether a single dose of doxycycline combined with the standard DEC regimen would reduce the incidence and severity of adverse reactions and increase the efficacy of standard treatment. Forty-four subjects from Tak Province were recruited into the randomized double-blind clinical trial study: 25 received DEC (300 mg) combined with a placebo, and 19 received DEC (300 mg) combined with doxycycline (200 mg). The incidences of adverse reactions to standard treatment were lower in the doxycycline group (45.5%) than in the placebo group (58.8%). Severe reactions occurred only in the placebo group (3 of 25 subjects). The severity of adverse reactions was significantly lower in the doxycycline group (mean score 0.45) than in the placebo group (mean score 1.17). The levels of IL-6 and Wolbachia DNA in the plasma were significantly lower in the doxycycline group. The filarial antigen levels were significantly lower in the doxycycline group at months 6 after treatment.}, } @article {pmid21062211, year = {2010}, author = {Farrokhi, S and Ashouri, A and Shirazi, J and Allahvari, H and Huigens, ME}, title = {A comparative study on the functional response of Wolbachia-infected and uninfected forms of the parasitoid wasp Trichogramma brassicae.}, journal = {Journal of insect science (Online)}, volume = {10}, number = {}, pages = {167}, pmid = {21062211}, issn = {1536-2442}, mesh = {Animals ; Host-Parasite Interactions/physiology ; Iran ; Logistic Models ; Models, Biological ; Moths/cytology/*parasitology ; Ovum/cytology ; Pest Control, Biological/*methods ; Sexual Behavior, Animal/*physiology ; Species Specificity ; Wasps/*microbiology/physiology ; *Wolbachia ; }, abstract = {Trichogramma species (Hymenoptera: Trichogrammatidae) are haplo-diploid egg parasitoids that are frequently used as biological control agents against lepidopteran pests. These wasps display two reproductive modes, including arrhenotoky (bisexuality) and thelytoky (unisexuality). Thelytokous forms are often associated with the presence of endosymbiotic Wolbachia bacteria. The use of thelytokous wasps has long been considered as a way to enhance the efficacy of biological control. The present study investigates the potential of a thelytokous Wolbachia-infected and an arrhenotokous uninfected Trichogramma brassicae Bezdenko strain as inundative biocontrol agents by evaluating their functional response towards different egg densities of the factitious host, the Angoumois grain moth, Sitotroga cerealella (Olivier) (Lepidoptera: Gelechiidae). The results revealed a type II functional response for both strains in which parasitism efficiency decreases with host egg density because of an increasing host handling time. A model with an indicator variable was used to compare the parameters of Holling's disc equation in different data sets. It was demonstrated that the two strains did not differ in host attack rate. However, the Wolbachia-infected strain did have an increased host handling time when compared to the bisexual strain. Some applied aspects of the findings are discussed.}, } @article {pmid21061989, year = {2010}, author = {Ahmed, MZ and Ren, SX and Mandour, NS and Greeff, JM and Qiu, BL}, title = {Prevalence of Wolbachia supergroups A and B in Bemisia tabaci (Hemiptera: Aleyrodidae) and some of its natural enemies.}, journal = {Journal of economic entomology}, volume = {103}, number = {5}, pages = {1848-1859}, doi = {10.1603/ec10033}, pmid = {21061989}, issn = {0022-0493}, mesh = {Animals ; Asia ; Hemiptera/*microbiology ; Mediterranean Region ; Middle East ; Pest Control/methods ; Plant Diseases/microbiology ; Plants/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia, a bacterial symbiont, is maternally transmitted in arthropods and nematodes. We report a systematic survey of Wolbachia taxonomy in the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), and in some of its natural enemies. For the first time, Wolbachia infections in B. tabaci are correlated with various whitefly genetic groups, host plants, and natural enemies as well as with geographical regions. Polymerase chain reaction using 16S rDNA and fisZ genes revealed two Wolbachia supergroups, A and B, exist as single or double infections in B. tabaci as well as in some of its aphelinid parasitoids and predatory beetles. Approximately 89% of B. tabaci sampled were infected by Wolbachia, among which 34% were infected by A, 51% were infected by B, and 5% were infected by both A and B supergroups. These infection frequencies differed among B. tabaci genetic groups and locations. The invasive B. tabaci genetic group from the Middle East Asia Minor 1 (also referred as B biotype) and Mediterranean (also referred as Q biotype) was more likely to harbor A than B, whereas native genetic groups in AsiaI and AsiaII were more likely to harbor B than A. Although 60% of aphelinid parasitoids and 72% of coccinellid beetles also were infected by Wolbachia, they were more likely to host B than A. Furthermore, for the first time we report Wolbachia in B biotype from specimens collected outside of China. Construction of a phylogenetic tree clearly indicated that the Wolbachia sequences from different genetic groups of B. tabaci were not only similar to each other but also to sequences from beetles and parasitoids, which may provide evidence of coevolution and horizontal transmission of Wolbachia populations.}, } @article {pmid21050430, year = {2010}, author = {Rodriguero, MS and Lanteri, AA and Confalonieri, VA}, title = {Mito-nuclear genetic comparison in a Wolbachia infected weevil: insights on reproductive mode, infection age and evolutionary forces shaping genetic variation.}, journal = {BMC evolutionary biology}, volume = {10}, number = {}, pages = {340}, pmid = {21050430}, issn = {1471-2148}, mesh = {Animals ; DNA, Mitochondrial/genetics ; *Evolution, Molecular ; Female ; Genetic Variation/*genetics ; Male ; Phylogeny ; Weevils/classification/*genetics/*microbiology ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: Maternally inherited endosymbionts like Wolbachia pipientis are in linkage disequilibrium with the mtDNA of their hosts. Therefore, they can induce selective sweeps, decreasing genetic diversity over many generations. This sex ratio distorter, that is involved in the origin of parthenogenesis and other reproductive alterations, infects the parthenogenetic weevil Naupactus cervinus, a serious pest of ornamental and fruit plants.

RESULTS: Molecular evolution analyses of mitochondrial (COI) and nuclear (ITS1) sequences from 309 individuals of Naupactus cervinus sampled over a broad range of its geographical distribution were carried out. Our results demonstrate lack of recombination in the nuclear fragment, non-random association between nuclear and mitochondrial genomes and the consequent coevolution of both genomes, being an indirect evidence of apomixis. This weevil is infected by a single Wolbachia strain, which could have caused a moderate bottleneck in the invaded population which survived the initial infection.

CONCLUSIONS: Clonal reproduction and Wolbachia infection induce the coevolution of bacterial, mitochondrial and nuclear genomes. The time elapsed since the Wolbachia invasion would have erased the traces of the demographic crash in the mtDNA, being the nuclear genome the only one that retained the signal of the bottleneck. The amount of genetic change accumulated in the mtDNA and the high prevalence of Wolbachia in all populations of N. cervinus agree with the hypothesis of an ancient infection. Wolbachia probably had great influence in shaping the genetic diversity of N. cervinus. However, it would have not caused the extinction of males, since sexual and asexual infected lineages coexisted until recent times.}, } @article {pmid21047535, year = {2011}, author = {Iturbe-Ormaetxe, I and Woolfit, M and Rancès, E and Duplouy, A and O'Neill, SL}, title = {A simple protocol to obtain highly pure Wolbachia endosymbiont DNA for genome sequencing.}, journal = {Journal of microbiological methods}, volume = {84}, number = {1}, pages = {134-136}, doi = {10.1016/j.mimet.2010.10.019}, pmid = {21047535}, issn = {1872-8359}, mesh = {Animals ; Culicidae/microbiology ; DNA, Bacterial/*isolation & purification ; Diptera/microbiology ; *Genome, Bacterial ; Molecular Biology/*methods ; Sequence Analysis, DNA ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Most genome sequencing projects using intracellular bacteria face difficulties in obtaining sufficient bacterial DNA free of host contamination. We have developed a simple and rapid protocol to isolate endosymbiont DNA virtually free from fly and mosquito host DNA. We purified DNA from six Wolbachia strains in preparation for genome sequencing using this method, and achieved up to 97% pure Wolbachia sequence, even after using frozen insects. This is a significant improvement for future Wolbachia and other endosymbiont genome projects.}, } @article {pmid21040533, year = {2010}, author = {Sirviö, A and Pamilo, P}, title = {Multiple endosymbionts in populations of the ant Formica cinerea.}, journal = {BMC evolutionary biology}, volume = {10}, number = {}, pages = {335}, pmid = {21040533}, issn = {1471-2148}, mesh = {Animals ; Ants/genetics/*microbiology/physiology ; Female ; Male ; Microsatellite Repeats/genetics ; Serratia/genetics/growth & development ; Social Behavior ; Symbiosis/genetics/*physiology ; Wolbachia/genetics/growth & development ; }, abstract = {BACKGROUND: Many insects, including ants, are infected by maternally inherited Wolbachia endosymbiotic bacteria though other secondary endosymbionts have not been reported in ants. It has been suggested that the ability of Wolbachia to invade and remain in an ant population depends on the number of coexisting queens in a colony. We study the genetic and social structure of populations in the ant Formica cinerea which is known to have populations with either monogynous or polygynous colonies. We screen populations for several endosymbiotic bacteria to evaluate the presence of different endosymbionts, possible association between their prevalence and the social structure, and the association between endosymbiont prevalence and genetic differentiation of ant populations.

RESULTS: We found three endosymbiotic bacteria; 19% of the nests were infected by Wolbachia, 3.8% by Cardinium and 33% by Serratia. There was significant variation among the populations regarding the proportion of nests infected by Serratia, Wolbachia and the pooled set of all the endosymbionts. Some individuals and colonies carried two of the bacteria, the frequency of double infections agreeing with the random expectation. The proportion of infected ants (individuals or colonies) did not correlate significantly with the population level relatedness values. The difference in the prevalence of Wolbachia between population pairs correlated significantly with the genetic distance (microsatellites) of the populations.

CONCLUSIONS: The discovery of several endosymbionts and co-infections by Wolbachia and Cardinium demonstrate the importance of screening several endosymbionts when evaluating their possible effects on social life and queen-worker conflicts over sex allocation. The low prevalence of Wolbachia in F. cinerea departs from the pattern observed in many other Formica ants in which all workers have been infected. It is likely that the strain of Wolbachia in F. cinerea differs from those in other Formica species. The correlation between the difference in Wolbachia prevalence and the pair-wise genetic distance of populations suggests that spreading of the bacteria is restricted by the isolation of the host populations.}, } @article {pmid21039221, year = {2010}, author = {Kazura, JW}, title = {Higher-dose, more frequent treatment of Wuchereria bancrofti.}, journal = {Clinical infectious diseases : an official publication of the Infectious Diseases Society of America}, volume = {51}, number = {11}, pages = {1236-1237}, doi = {10.1086/657064}, pmid = {21039221}, issn = {1537-6591}, mesh = {Albendazole/*administration & dosage ; Animals ; Drug Therapy, Combination/methods ; Filariasis/*drug therapy/*epidemiology/transmission ; Filaricides/*administration & dosage ; Humans ; Ivermectin/*administration & dosage ; Parasitemia/drug therapy ; Symbiosis ; Wolbachia/drug effects/physiology ; Wuchereria bancrofti/*drug effects/*isolation & purification ; }, } @article {pmid21033053, year = {2010}, author = {Wiwatanaratanabutr, I and Allan, S and Linthicum, K and Kittayapong, P}, title = {Strain-specific differences in mating, oviposition, and host-seeking behavior between Wolbachia-infected and uninfected Aedes albopictus.}, journal = {Journal of the American Mosquito Control Association}, volume = {26}, number = {3}, pages = {265-273}, doi = {10.2987/09-5937.1}, pmid = {21033053}, issn = {8756-971X}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Carbon Dioxide ; Feeding Behavior/*physiology ; Female ; Humans ; Male ; Methylene Chloride ; Oviposition/*physiology ; Sexual Behavior, Animal/*physiology ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia are maternally inherited bacteria that cause various reproductive alterations in their arthropod hosts, including cytoplasmic incompatibility. In this study, we compared mating, oviposition, and host-seeking behavior of Wolbachia-infected (Houston [HOU], Gainesville [GNV]) and Houston uninfected (HT1) Aedes albopictus. In mating assays with virgin mosquitoes, mating success of Wolbachia-infected males was significantly higher than uninfected strains. Mating success was highest with HOU males exposed to infected (95%) and uninfected females (100%), and lowest with HT1 males exposed to infected (20%) and uninfected (25%) females. Results suggested that Wolbachia infection may influence the reproductive behavior of this mosquito. There were no clear differences in oviposition responses between strains, with all strains ovipositing significantly more often on hay infusion and larval rearing water than on water controls and least frequently on 4-methylphenol. Strains of Ae. albopictus females were host-seeking a human when given a choice. Responses to a human arm, acetone, CO2, and dichloromethane were generally higher from the Houston strains than from the GNV strain. Responses of HOU and HT1 females differed from GNV with greater responses to the arm and CO2.}, } @article {pmid20981548, year = {2011}, author = {Ahantarig, A and Chauvatcharin, N and Ruang-areerate, T and Baimai, V and Kittayapong, P}, title = {Infection incidence and relative density of the bacteriophage WO-B in Aedes albopictus mosquitoes from fields in Thailand.}, journal = {Current microbiology}, volume = {62}, number = {3}, pages = {816-820}, pmid = {20981548}, issn = {1432-0991}, mesh = {Aedes/*microbiology/virology ; Animals ; Bacteriophages/*isolation & purification ; DNA, Viral/genetics/isolation & purification ; Female ; Geography ; Thailand ; Wolbachia/*virology ; }, abstract = {We have used real-time quantitative PCR to measure, for the first time, the relative phage WO-B orf7 density and infection incidence in Aedes albopictus mosquitoes from fields in Thailand. Our results showed that the infection incidence of phage WO-B in this mosquito, sampled from geographically different places in Thailand, was 97.9%. Average relative densities of the offspring were different when collected from diverse parts and reared under the same conditions in the laboratory. Our results also revealed that geographical differences within Thailand did not influence the maternal transmission rate of bacteriophage WO-B. In addition, the orf7 loci might not be strictly associated with Wolbachia, because less than 100% of them were maternally inherited. This discovery does not support the hypothesis that bacteriophage WO-B is involved in Aedes albopictus' cytoplasmic incompatibility. Whether this bacteriophage actually is involved in Wolbachia-induced cytoplasmic incompatibility in this mosquito thus needs further investigation, and additional densities of phage WO-B loci should be integrated.}, } @article {pmid20976219, year = {2010}, author = {Frentiu, FD and Robinson, J and Young, PR and McGraw, EA and O'Neill, SL}, title = {Wolbachia-mediated resistance to dengue virus infection and death at the cellular level.}, journal = {PloS one}, volume = {5}, number = {10}, pages = {e13398}, pmid = {20976219}, issn = {1932-6203}, mesh = {Aedes/virology ; Animals ; *Cell Death ; Cell Line ; Cell Proliferation ; Dengue Virus/*pathogenicity/physiology ; Insect Vectors ; Virus Replication ; Wolbachia/cytology/*immunology/virology ; }, abstract = {BACKGROUND: Dengue is currently the most important arthropod-borne viral disease of humans. Recent work has shown dengue virus displays limited replication in its primary vector, the mosquito Aedes aegypti, when the insect harbors the endosymbiotic bacterium Wolbachia pipientis. Wolbachia-mediated inhibition of virus replication may lead to novel methods of arboviral control, yet the functional and cellular mechanisms that underpin it are unknown.

Using paired Wolbachia-infected and uninfected Aedes-derived cell lines and dengue virus, we confirm the phenomenon of viral inhibition at the cellular level. Although Wolbachia imposes a fitness cost to cells via reduced proliferation, it also provides a significant degree of protection from virus-induced mortality. The extent of viral inhibition is related to the density of Wolbachia per cell, with highly infected cell lines showing almost complete protection from dengue infection and dramatically reduced virus titers compared to lines not infected with the bacteria.

CONCLUSIONS/SIGNIFICANCE: We have shown that cells infected with Wolbachia display inhibition of dengue virus replication, that the extent of inhibition is related to bacterial density and that Wolbachia infection, although costly, will provide a fitness benefit in some circumstances. Our results parallel findings in mosquitoes and flies, indicating that cell line models will provide useful and experimentally tractable models to study the mechanisms underlying Wolbachia-mediated protection from viruses.}, } @article {pmid20949416, year = {2010}, author = {Frank, K and Heald, RD}, title = {The emerging role of Wolbachia species in heartworm disease.}, journal = {Compendium (Yardley, PA)}, volume = {32}, number = {4}, pages = {E4}, pmid = {20949416}, issn = {1940-8315}, mesh = {Animals ; Anti-Bacterial Agents/adverse effects/therapeutic use ; Cat Diseases/drug therapy/*microbiology ; Cats ; Dirofilaria immitis/*microbiology ; Dirofilariasis/*microbiology ; Dog Diseases/drug therapy/*microbiology ; Dogs ; Wolbachia/drug effects/*pathogenicity ; }, abstract = {Heartworm disease was first recognized in dogs more than 100 years ago and is still prevalent among dogs and found in cats worldwide. The complications of heartworm disease can be devastating, and treatment carries risks. Wolbachia spp are gram-negative bacteria that infect filarial nematodes, including Dirofilaria immitis, and elicit an inflammatory response in cats and dogs. Antimicrobial therapy directed against these bacteria has resulted in decreased microfilarial loads, inhibition of the development of larval worms, female worm infertility, and reduced numbers of Wolbachia organisms. Antimicrobial therapy against Wolbachia spp may be useful in treating heartworm disease in cats and dogs, but further research is needed.}, } @article {pmid20949079, year = {2010}, author = {Kambris, Z and Blagborough, AM and Pinto, SB and Blagrove, MS and Godfray, HC and Sinden, RE and Sinkins, SP}, title = {Wolbachia stimulates immune gene expression and inhibits plasmodium development in Anopheles gambiae.}, journal = {PLoS pathogens}, volume = {6}, number = {10}, pages = {e1001143}, pmid = {20949079}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; 079059/WT_/Wellcome Trust/United Kingdom ; LDAD_P15820/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Animals, Genetically Modified ; Anopheles/*immunology/microbiology/parasitology ; Antibiosis/genetics/immunology ; Communicable Disease Control/methods ; Female ; Gene Expression Regulation/immunology ; Gene Knockdown Techniques ; Host-Parasite Interactions/genetics/immunology ; Immunity, Innate/*genetics ; Insect Proteins/antagonists & inhibitors/genetics/physiology ; Malaria/immunology/prevention & control ; Plasmodium/*growth & development/*immunology/pathogenicity ; Up-Regulation/genetics ; Wolbachia/genetics/*physiology ; }, abstract = {The over-replicating wMelPop strain of the endosymbiont Wolbachia pipientis has recently been shown to be capable of inducing immune upregulation and inhibition of pathogen transmission in Aedes aegypti mosquitoes. In order to examine whether comparable effects would be seen in the malaria vector Anopheles gambiae, transient somatic infections of wMelPop were created by intrathoracic inoculation. Upregulation of six selected immune genes was observed compared to controls, at least two of which (LRIM1 and TEP1) influence the development of malaria parasites. A stably infected An. gambiae cell line also showed increased expression of malaria-related immune genes. Highly significant reductions in Plasmodium infection intensity were observed in the wMelPop-infected cohort, and using gene knockdown, evidence for the role of TEP1 in this phenotype was obtained. Comparing the levels of upregulation in somatic and stably inherited wMelPop infections in Ae. aegypti revealed that levels of upregulation were lower in the somatic infections than in the stably transinfected line; inhibition of development of Brugia filarial nematodes was nevertheless observed in the somatic wMelPop infected females. Thus we consider that the effects observed in An. gambiae are also likely to be more pronounced if stably inherited wMelPop transinfections can be created, and that somatic infections of Wolbachia provide a useful model for examining effects on pathogen development or dissemination. The data are discussed with respect to the comparative effects on malaria vectorial capacity of life shortening and direct inhibition of Plasmodium development that can be produced by Wolbachia.}, } @article {pmid20946650, year = {2010}, author = {Johnston, KL and Wu, B and Guimarães, A and Ford, L and Slatko, BE and Taylor, MJ}, title = {Lipoprotein biosynthesis as a target for anti-Wolbachia treatment of filarial nematodes.}, journal = {Parasites & vectors}, volume = {3}, number = {}, pages = {99}, pmid = {20946650}, issn = {1756-3305}, abstract = {BACKGROUND: Lymphatic filariasis and onchocerciasis are debilitating diseases caused by filarial nematodes. Disease pathogenesis is induced by inflammatory responses following the death of the parasite. Wolbachia endosymbionts of filariae are potent inducers of innate and adaptive inflammation and bacterial lipoproteins have been identified as the ligands that bind toll-like receptors (TLR) 2 and TLR6. Lipoproteins are important structural and functional components of bacteria and therefore enzymes involved in Wolbachia lipoprotein biosynthesis are potential chemotherapeutic targets.

RESULTS: Globomycin, a signal peptidase II (LspA) inhibitor, has activity against Gram-negative bacteria and a putative lspA gene has been identified from the Wolbachia genome of Brugia malayi (wBm). The amino acids required for function are strictly conserved and functionality was verified by complementation tests in a temperature-sensitive Escherichia coli lspA mutant. Also, transformation of wild type E. coli with Wolbachia lspA conferred significant globomycin resistance. A cell-based screen has been developed utilizing a Wolbachia-containing Aedes albopictus cell line to assay novel compounds active against Wolbachia. Globomycin was screened using this assay, which resulted in a dose-dependent reduction in Wolbachia load. Furthermore, globomycin was also effective in reducing the motility and viability of adult B. malayi in vitro.

CONCLUSIONS: These studies validate lipoprotein biosynthesis as a target in an organism for which no genetic tools are available. Further studies to evaluate drugs targeting this pathway are underway as part of the A-WOL drug discovery and development program.}, } @article {pmid20944019, year = {2011}, author = {Chafee, ME and Zecher, CN and Gourley, ML and Schmidt, VT and Chen, JH and Bordenstein, SR and Clark, ME and Bordenstein, SR}, title = {Decoupling of host-symbiont-phage coadaptations following transfer between insect species.}, journal = {Genetics}, volume = {187}, number = {1}, pages = {203-215}, pmid = {20944019}, issn = {1943-2631}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {*Adaptation, Physiological/genetics ; Animals ; Bacteriophages/metabolism/*physiology ; Cytoplasm/metabolism/microbiology/virology ; Female ; Hymenoptera/genetics/microbiology/*physiology/*virology ; Male ; Sexual Behavior, Animal/physiology ; Species Specificity ; Starvation/genetics/microbiology/virology ; Symbiosis/genetics/*physiology ; Virion/metabolism ; Wolbachia/*physiology ; }, abstract = {Transferring endosymbiotic bacteria between different host species can perturb the coordinated regulation of the host and bacterial genomes. Here we use the most common maternally transmitted bacteria, Wolbachia pipientis, to test the consequences of host genetic background on infection densities and the processes underlying those changes in the parasitoid wasp genus Nasonia. Introgressing the genome of Nasonia giraulti into the infected cytoplasm of N. vitripennis causes a two-order-of-magnitude increase in bacterial loads in adults and a proliferation of the infection to somatic tissues. The host effect on W. pipientis distribution and densities is associated with a twofold decrease in densities of the temperate phage WO-B. Returning the bacteria from the new host species back to the resident host species restores the bacteria and phage to their native densities. To our knowledge, this is the first study to report a host-microbe genetic interaction that affects the densities of both W. pipientis and bacteriophage WO-B. The consequences of the increased bacterial density include a reduction in fecundity, an increase in levels of cytoplasmic incompatibility (CI), and unexpectedly, male-to-female transfer of the bacteria to uninfected females and an increased acceptance of densely infected females to interspecific mates. While paternal inheritance of the W. pipientis was not observed, the high incidence of male-to-female transfer in the introgressed background raises the possibility that paternal transmission could be more likely in hybrids where paternal leakage of other cytoplasmic elements is also known to occur. Taken together, these results establish a major change in W. pipientis densities and tissue tropism between closely related species and support a model in which phage WO, Wolbachia, and arthropods form a tripartite symbiotic association in which all three are integral to understanding the biology of this widespread endosymbiosis.}, } @article {pmid20939837, year = {2010}, author = {Kobayashi, Y and Telschow, A}, title = {Cytoplasmic feminizing elements in a two-population model: infection dynamics, gene flow modification, and the spread of autosomal suppressors.}, journal = {Journal of evolutionary biology}, volume = {23}, number = {12}, pages = {2558-2568}, doi = {10.1111/j.1420-9101.2010.02116.x}, pmid = {20939837}, issn = {1420-9101}, mesh = {Animals ; Biological Evolution ; Female ; *Gene Flow ; Host-Parasite Interactions/*genetics ; Male ; *Models, Genetic ; Population Dynamics ; Sex Chromosomes ; *Sex Determination Processes ; Sex Ratio ; Wolbachia/physiology ; }, abstract = {We investigate the dynamics of a cytoplasmic parasitic element with feminizing effect in a two-population model. We assume that the host species has a ZZ/ZW sex determination system. Our analysis reveals that the feminizer and the W chromosome can stably coexist by dominating different populations if the transmission rate differs significantly between the populations and migration is sufficiently weak. In the equilibrium of coexistence, genetic influx at any host autosomal locus is strongly enhanced in the population where infection is prevalent but not modified in the other population. We further explore conditions for the spread of autosomal suppressor genes that reduce transmission of feminizing elements to the cost of host viability, and compute their equilibrium frequencies. Our results confirm the hypothesis that selfish genetic elements convert infected host populations into genetic sinks, thereby restricting the spread of transmission suppressors.}, } @article {pmid20889885, year = {2010}, author = {Bennuru, S and Maldarelli, G and Kumaraswami, V and Klion, AD and Nutman, TB}, title = {Elevated levels of plasma angiogenic factors are associated with human lymphatic filarial infections.}, journal = {The American journal of tropical medicine and hygiene}, volume = {83}, number = {4}, pages = {884-890}, pmid = {20889885}, issn = {1476-1645}, mesh = {Adolescent ; Adult ; Aged ; Angiogenic Proteins/*blood/metabolism ; Angiopoietins/blood/metabolism ; Animals ; Doxycycline/therapeutic use ; Elephantiasis, Filarial/*blood/drug therapy/epidemiology/parasitology ; Fibroblast Growth Factor 2/blood/metabolism ; Filaricides/therapeutic use ; Humans ; India/epidemiology ; Mali/epidemiology ; Middle Aged ; Vascular Endothelial Growth Factors/blood/metabolism ; *Wuchereria bancrofti ; Young Adult ; }, abstract = {Lymphatic dilatation, dysfunction, and lymphangiogenesis are hallmarks of patent lymphatic filariasis, observed even in those with subclinical microfilaremia, through processes associated, in part, by vascular endothelial growth factors (VEGFs). A panel of pro-angiogenic factors was measured in the plasma of subjects from filaria-endemic regions using multiplexed immunological assays. Compared with endemic normal control subjects, those with both subclinical microfilaremia, and those with longstanding lymphedema had significantly elevated levels of VEGF-A, VEGF-C, VEGF-D, and angiopoietins (Ang-1/Ang-2), with only levels of basic fibroblast growth factor (bFGF) and placental growth factor (PlGF) being elevated only if lymphedema was evident. Furthermore, levels of these factors 1-year post-treatment with doxycycline were similar to pretreatment levels suggesting a minimal role, if any, for Wolbachia. Our data support the concept that filarial infection per se is associated with elevated levels of most of the known pro-angiogenic factors, with only a few being associated with the serious pathologic consequences associated with Wuchereria bancrofti infection.}, } @article {pmid20880864, year = {2011}, author = {Lewis, Z and Champion de Crespigny, FE and Sait, SM and Tregenza, T and Wedell, N}, title = {Wolbachia infection lowers fertile sperm transfer in a moth.}, journal = {Biology letters}, volume = {7}, number = {2}, pages = {187-189}, pmid = {20880864}, issn = {1744-957X}, mesh = {Animals ; Female ; Fertility/physiology ; Male ; Moths/*microbiology/physiology ; Spermatozoa/*microbiology ; Wolbachia/*isolation & purification ; }, abstract = {The endosymbiotic bacterium Wolbachia pipientis manipulates host reproduction by rendering infected males reproductively incompatible with uninfected females (cytoplasmic incompatibility; CI). CI is believed to occur as a result of Wolbachia-induced modifications to sperm during maturation, which prevent infected sperm from initiating successful zygote development when fertilizing uninfected females' eggs. However, the mechanism by which CI occurs has been little studied outside the genus Drosophila. Here, we show that in the sperm heteromorphic Mediterranean flour moth, Ephestia kuehniella, infected males transfer fewer fertile sperm at mating than uninfected males. In contrast, non-fertile apyrene sperm are not affected. This indicates that Wolbachia may only affect fertile sperm production and highlights the potential of the Lepidoptera as a model for examining the mechanism by which Wolbachia induces CI in insects.}, } @article {pmid20859003, year = {2010}, author = {Sakamoto, L and Ichikawa, Y and Sakata, Y and Matsumoto, K and Inokuma, H}, title = {Detection of Anaplasma bovis DNA in the peripheral blood of domestic dogs in Japan.}, journal = {Japanese journal of infectious diseases}, volume = {63}, number = {5}, pages = {349-352}, pmid = {20859003}, issn = {1884-2836}, mesh = {Anaplasma/*genetics/isolation & purification ; Anaplasmosis/epidemiology/*microbiology ; Animals ; Cross-Sectional Studies ; DNA, Bacterial/*blood ; Dog Diseases/blood/epidemiology/*microbiology ; Dogs ; Ehrlichia/genetics/isolation & purification ; Ehrlichiosis/epidemiology/microbiology/veterinary ; Geography ; Japan/epidemiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {The prevalence of Ehrlichia and Anaplasma in 1,427 dogs from 32 Japanese prefectures was evaluated by PCR and DNA nucleotide sequencing. PCR screening demonstrated that 18 dogs (1.3%) were positive for Anaplasmataceae. Sequence analysis revealed that 14 of the amplicons were most closely related to Wolbachia spp., symbionts of Dirofilaria immitis, whereas three were identified as Anaplasma bovis. The remaining amplicon could not be sequenced. Almost the entire sequence of 16S rRNA (1,452 bp) from one of the positive specimens was determined, and subsequent phylogenetic analysis confirmed that the detected sequence was that of A. bovis. This is the first detection of A. bovis DNA fragments in dogs. Species-specific nested PCR showed that 15 (1.1%) of the 1,427 dogs involved in this study were positive for A. bovis. The geographical distribution of these dogs ranged from Aomori Prefecture in northern Japan to Kagoshima Prefecture in the south. The relationship between A. bovis infection and clinical disease is not yet clearly understood.}, } @article {pmid20855945, year = {2010}, author = {Blatch, SA and Meyer, KW and Harrison, JF}, title = {Effects of dietary folic acid level and symbiotic folate production on fitness and development in the fruit fly Drosophila melanogaster.}, journal = {Fly}, volume = {4}, number = {4}, pages = {312-319}, doi = {10.4161/fly.4.4.13258}, pmid = {20855945}, issn = {1933-6942}, mesh = {Animals ; Drosophila melanogaster/*drug effects/growth & development/microbiology ; Folic Acid/biosynthesis/*pharmacology ; Larva/drug effects/growth & development/microbiology ; Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {Folic acid is a vitamin for probably all animals. When converted to folate forms, it is used in DNA synthesis and amino acid metabolism. Literature suggests insects must consume folates, folates do not affect others, is a toxin for some, and that a few insects synthesize it. It has been reported that Drosophila melanogaster does not consistently need dietary folate because it can synthesize it. This seems unlikely since animals generally lack this ability. More likely, folates thought to have been made by the fly came from microbial symbionts. We aimed to clarify how dietary folic acid affects fitness and development in fruit flies and whether flies may receive folates from microbial symbionts. We found larvae were more viable and developed faster with increasing dietary folic acid, with the surprising exception that larvae fed nearly-zero folic acid developed faster. Their body folate levels did not significantly differ from those that consumed up to 600 times more folic acid. However, these flies fed little folate only achieved normal body folate levels and development times when antibiotics were excluded from the diet. When flies consumed near-zero folates with antibiotics, their body folate levels decreased and development was prolonged. An assay for the endosymbiont Wolbachia in flies used to generate the experimental flies did not show presence of these bacteria. Our data suggest D. melanogaster can harbor unknown bacterial symbiont(s) that provide essential folates to their host when it is scarce in the diet, allowing the fruit fly to maintain growth and development.}, } @article {pmid20854481, year = {2011}, author = {Yamada, R and Iturbe-Ormaetxe, I and Brownlie, JC and O'Neill, SL}, title = {Functional test of the influence of Wolbachia genes on cytoplasmic incompatibility expression in Drosophila melanogaster.}, journal = {Insect molecular biology}, volume = {20}, number = {1}, pages = {75-85}, doi = {10.1111/j.1365-2583.2010.01042.x}, pmid = {20854481}, issn = {1365-2583}, mesh = {Animals ; Animals, Genetically Modified/genetics/metabolism/microbiology ; Cytoplasm/*genetics/microbiology ; Drosophila melanogaster/cytology/*genetics/metabolism/*microbiology ; Female ; Gene Expression Regulation ; Genes, Insect ; *Genetic Techniques ; Male ; Molecular Sequence Data ; Wolbachia/*genetics/physiology ; }, abstract = {Wolbachia are inherited intracellular bacteria that infect a broad range of invertebrate hosts. They commonly manipulate host reproduction in a variety of ways and thereby favour their invasion into host populations. While the biology of Wolbachia has been extensively studied at the ecological and phenotypic level, little is known about the molecular mechanisms underlying the interaction between Wolbachia and their hosts. Recent comparative genomics studies of Wolbachia strains have revealed putative candidate genes involved in the expression of cytoplasmic incompatibility (CI) in insects. However the functional testing of these genes is hindered by the lack of available genetic tools in Wolbachia. To circumvent this problem we generated transgenic Drosophila lines expressing various Wolbachia CI candidate genes under the control of the GAL4/UAS system in order to evaluate their possible role in Wolbachia-related phenotypes in Drosophila. The expression of a number of these genes in Drosophila melanogaster failed to mimic or alter CI phenotypes across a range of Wolbachia backgrounds or in the absence of Wolbachia.}, } @article {pmid20850932, year = {2010}, author = {Neary, JM and Trees, AJ and Ekale, DD and Tanya, VN and Hetzel, U and Makepeace, BL}, title = {Onchocerca armillata contains the endosymbiotic bacterium Wolbachia and elicits a limited inflammatory response.}, journal = {Veterinary parasitology}, volume = {174}, number = {3-4}, pages = {267-276}, pmid = {20850932}, issn = {1873-2550}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aging ; Animals ; Aorta/pathology ; Cattle ; Cattle Diseases/parasitology/pathology ; Female ; Inflammation/parasitology/pathology/*veterinary ; Male ; Onchocerca/*microbiology ; Onchocerciasis/parasitology/pathology/veterinary ; Sex Characteristics ; Wolbachia/*physiology ; }, abstract = {Human onchocerciasis, also known as River Blindness, is a debilitating disease caused by the filarial nematode Onchocerca volvulus. Many, but not all, filarial nematodes carry within their tissues endosymbiotic, Rickettsia-like bacteria of the genus Wolbachia. Onchocerca spp. infections in cattle offer the most relevant, analogous host-parasite model system. West African cattle are commonly co-infected with four Onchocerca spp.; two of these are Wolbachia-positive (Onchocerca gutturosa and Onchocerca ochengi), and the remainder are of unknown Wolbachia status (Onchocerca dukei and Onchocerca armillata). Previous studies have suggested that worm survival is dependent on this bacterium. O. armillata, an abundant parasite of African cattle that has received little attention, is a primitive species that may lack Wolbachia. The objectives of this study were to determine if O. armillata carries Wolbachia and to provide preliminary descriptions of the host inflammatory cell environment around the adult worms. The findings may support or refute the hypothesis that a prime contribution of Wolbachia is to permit long-term survival and reproduction of certain Onchocerca spp. (including O. volvulus in humans). O. armillata adult worms were found in the aorta of 90.7% of cattle (n=54) slaughtered at an abattoir in Ngaoundéré, Adamawa Region, Cameroon. The presence of Wolbachia in O. armillata was confirmed by a specific anti-Wolbachia surface protein antibody detected using a peroxidase conjugate (immunohistochemistry) and PCR for detection of Wolbachia-specific sequences within DNA extracts from frozen worms. Tissue sections stained with haematoxylin and eosin showed the host cell response to be dominated by macrophages and fibroblasts. This is unusual compared with nodule-dwelling Wolbachia-positive Onchocerca spp., where the host response is typically characterised by granulocytes, and suggests that the mechanisms for worm survival employed by this species (which is probably motile) may differ.}, } @article {pmid20847694, year = {2010}, author = {Bockarie, MJ and Deb, RM}, title = {Elimination of lymphatic filariasis: do we have the drugs to complete the job?.}, journal = {Current opinion in infectious diseases}, volume = {23}, number = {6}, pages = {617-620}, doi = {10.1097/QCO.0b013e32833fdee5}, pmid = {20847694}, issn = {1473-6527}, mesh = {Africa ; Animals ; Antiparasitic Agents/pharmacology/*therapeutic use ; Drug Administration Schedule ; Drug Therapy, Combination ; Elephantiasis, Filarial/complications/*drug therapy/transmission ; Filaricides/administration & dosage/pharmacology/*therapeutic use ; Global Health ; Humans ; Insect Vectors/parasitology ; Loiasis/complications ; Program Evaluation ; Time Factors ; Treatment Outcome ; World Health Organization ; }, abstract = {PURPOSE OF REVIEW: Lymphatic filariasis is targeted for elimination globally through mass drug administration (MDA) with diethylcarbamazine or ivermectin monotherapy, or either drug in combination with albendazole. However, many countries that have implemented MDA annually for over 5 years are yet to interrupt transmission. This review describes the current drugs used in MDA and highlights the challenges facing the WHO Global Programme to Eliminate Lymphatic Filariasis (GPELF).

RECENT FINDINGS: Current drugs used for MDA implementation by national elimination programmes only temporarily clear microfilariae without killing all adult worms. Generally, reports of serious adverse events associated with MDA for lymphatic filariasis using current drugs are uncommon. However, in areas in Africa where lymphatic filariasis co-exists with Loa loa, progressive neurologic decline and encephalopathy within a few days of taking ivermectin have caused great concern. Doxycycline, which is effective at eliminating the Wolbachia symbiont from the lymphatic filariasis parasite, is showing promise as an alternative treatment option for areas where lymphatic filariasis is co-endemic with Loa loa.

SUMMARY: Alternative and effective MDA regimens and strategies will be needed if the GPELF is to achieve the goals of global elimination of lymphatic filariasis by 2020. Further research to test new drug regimens (including single high doses of albendazole) or alternative treatment regimens (including biannual treatment schedules) may also be necessary. A new drug, moxidectin, which is currently under development for use against onchocerciasis, may be effective against lymphatic filariasis.}, } @article {pmid20819906, year = {2011}, author = {Leclercq, S and Giraud, I and Cordaux, R}, title = {Remarkable abundance and evolution of mobile group II introns in Wolbachia bacterial endosymbionts.}, journal = {Molecular biology and evolution}, volume = {28}, number = {1}, pages = {685-697}, doi = {10.1093/molbev/msq238}, pmid = {20819906}, issn = {1537-1719}, mesh = {Animals ; Base Sequence ; DNA Transposable Elements/*genetics ; *Evolution, Molecular ; Gene Transfer, Horizontal ; Genome, Bacterial ; *Introns ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Symbiosis/*genetics ; Wolbachia/classification/*genetics ; }, abstract = {The streamlined genomes of ancient obligate endosymbionts generally lack transposable elements, as a consequence of their intracellular confinement. Yet, the genomes of Wolbachia, one of the most abundant bacterial endosymbionts on Earth, are littered with transposable elements, in particular insertion sequences (ISs). This paradox raises the question of whether or not such a mobile DNA proliferation reflects a special feature of ISs. In this study, we focused on another class of transposable elements, group II introns, and conducted an in-depth analysis of their content and the microevolutionary processes responsible for their dynamics within Wolbachia genomes. We report an exceptionally high intron abundance and striking differences in copy numbers between Wolbachia strains as well as between intron families. Our bioinformatics and experimental results provide strong evidence that intron diversity is mainly caused by recent (and perhaps ongoing) mobility and horizontal transfers. Our data also support several temporally independent intron invasions during Wolbachia evolution. Furthermore, group II intron spread in some Wolbachia strains may be regulated through gene conversion-mediated inactivation of intron copies. Finally, we found introns to be involved in numerous genomic rearrangements. This underscores the high recombinogenic potential of group II introns, contrary to general expectations. Overall, our study represents the first comprehensive analysis of group II intron evolutionary dynamics in obligate intracellular bacteria. Our results show that bacterial endosymbionts with reduced genomes can sustain high loads of mobile group II introns, as hypothesized for the endosymbiont ancestor of mitochondria during early eukaryote evolution.}, } @article {pmid20817243, year = {2010}, author = {Sacchi, L and Genchi, M and Clementi, E and Negri, I and Alma, A and Ohler, S and Sassera, D and Bourtzis, K and Bandi, C}, title = {Bacteriocyte-like cells harbour Wolbachia in the ovary of Drosophila melanogaster (Insecta, Diptera) and Zyginidia pullula (Insecta, Hemiptera).}, journal = {Tissue & cell}, volume = {42}, number = {5}, pages = {328-333}, doi = {10.1016/j.tice.2010.07.009}, pmid = {20817243}, issn = {1532-3072}, mesh = {Animals ; Drosophila melanogaster/*microbiology ; Female ; Hemiptera/*microbiology ; Ovary/*cytology/microbiology ; Symbiosis ; Wolbachia ; }, abstract = {Wolbachia is the most widespread bacterial endosymbiont in insects. It is responsible for a variety of reproductive alterations of the hosts. Wolbachia is transmitted through the germline from mother to offspring and, in rare cases, between individuals. This implies that acquired properties (through symbiosis with Wolbachia) can become heritable. We investigated the transovarial inheritance of Wolbachia in two phylogenetically distant insects, Drosophila melanogaster and Zyginidia pullula. We detected in both systems bacteriocyte-like cells, densely packed with Wolbachia endosymbionts, at the tip of the ovarioles. Bacteriocytes are cells specialized to harbour bacteria, typical of mutualistic insect symbiosis. Our observations of bacteriocyte-like cells harbouring Wolbachia in the ovary emphasize the plasticity of the female reproductive system of insects, which maintains its function while some cells are densely colonized by bacteria. In summary, there is evidence from different insects that bacteria which behave as parasites of reproduction are harboured by cells resembling bacteriocytes, which appear to mediate transmission of the bacteria to the progeny. It seems a valid hypothesis that the bacteriocyte-like cells that we observed are not the result of a co-evolution of host and symbiont, considering that Wolbachia is not an obligatory symbiont in Drosophila and Zyginidia.}, } @article {pmid20810215, year = {2010}, author = {Rossi, MI and Paiva, J and Bendas, A and Mendes-de-Almeida, F and Knackfuss, F and Miranda, M and Guerrero, J and Fernandes, O and Labarthe, N}, title = {Effects of doxycycline on the endosymbiont Wolbachia in Dirofilaria immitis (Leidy, 1856)--naturally infected dogs.}, journal = {Veterinary parasitology}, volume = {174}, number = {1-2}, pages = {119-123}, doi = {10.1016/j.vetpar.2010.07.019}, pmid = {20810215}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Antigens, Bacterial/blood ; DNA, Bacterial/blood ; Dirofilaria immitis/drug effects/*microbiology ; Dog Diseases/*microbiology/*parasitology ; Dogs ; Doxycycline/*pharmacology ; Time Factors ; Wolbachia/*drug effects ; }, abstract = {Dirofilaria immitis carries intracellular endosymbiotic bacteria of the genus Wolbachia, known to be vital for the worms and sensitive to tetracycline antibiotics. With the purpose of studying the interaction between D. immitis and the endosymbiont Wolbachia sp., heartworm naturally infected microfilaremic or antigenemic dogs were treated with doxycycline (10mg/kg/day of the drug in three cycles of 21 days each, with 6-month intervals). Blood samples were collected on days 0, 7 and 21 of each treatment as well as on day 111 after the beginning of each cycle. A final sample was collected on day 723 from the beginning of the first treatment. The samples were examined for the presence and number of microfilariae and the presence of antigen as well as the presences of D. immitis and Wolbachia sp. DNA using PCR (polymerase chain reaction). With this approach, an evaluation of the effect of doxycycline on antigenemia and on the presence of Wolbachia sp. DNA in dogs with heartworm infection was possible. Doxycycline treatment did not alter the detection of adult parasite antigens with the exception of two animals, though the number of animals carrying Wolbachia sp. DNA decreased, despite the presence of the microfilariae. The effect of the antibiotic therapy on the worms may have interfered with the transmission of heartworm disease because the population of microfilariae and the number of microfilaremic dogs were reduced and the microfilariae positive samples that were found did not test positive for Wolbachia sp. in many cases. These findings suggest that in areas were doxycycline is extensively used D. immitis transmission may be impaired by the reduction on the number of microfilariae and on the endosymbiotic bacteria in the larvae turning them incapable of completing development once they infected a new host.}, } @article {pmid20809419, year = {2011}, author = {Brattig, NW and Racz, P and Hoerauf, A and Büttner, DW}, title = {Strong expression of TGF-beta in human host tissues around subcutaneous Dirofilaria repens.}, journal = {Parasitology research}, volume = {108}, number = {6}, pages = {1347-1354}, pmid = {20809419}, issn = {1432-1955}, mesh = {Animals ; Antibodies, Monoclonal ; B-Lymphocytes/metabolism ; Dirofilaria/*immunology ; Dirofilariasis/*immunology ; HLA-DR Antigens/biosynthesis ; Humans ; Lymph Nodes/cytology/pathology ; Lymphocytes/metabolism ; Lymphoid Tissue/metabolism/pathology ; Macrophages/metabolism ; Skin/parasitology ; Subcutaneous Tissue/*parasitology ; Transforming Growth Factor beta/biosynthesis/genetics/*immunology ; Wolbachia/pathogenicity ; }, abstract = {Dirofilaria repens and other Dirofilaria species are widely distributed parasitic nematodes of carnivores, which occasionally are transmitted to men, causing subcutaneous nodules. In humans, it usually occurs only as single male or female filariae without production of microfilariae. The non-productive living or dead Dirofilaria worms in subcutaneous biopsies from 15 human patients permitted us to study the role of the pleiotropic and immunoregulatory cytokine transforming growth factor beta (TGF-beta) independent from the influence of microfilariae. Antiserum against latent TGF-beta 1 was used for an immunohistological examination. In the infiltrates around female and male filariae, there occurred strongly TGF-beta-positive macrophages, mast cells, endothelial cells, fibrocytes, and giant cells adjacent to dead worms. In one nodule, secondary lymph follicles were observed with clearly TGF-beta-positive B cells in the mantle zone and weakly positive macrophages and B cells in the germinal centre. A network of CD35-positive follicular dendritic cells was observed in the germinal centre. All Dirofilaria contained Wolbachia endobacteria, which probably had attracted the numerous TGF-beta-negative neutrophils near to the worm. Wolbachia were phagocytosed by neutrophils adjacent to dead filariae. Macrophages and lymphocytes expressed the MHC class II molecule HLA-DR in small accumulations of immune cells in the outer zone of the infiltrate and the mantle zone and germinal centre of secondary lymph follicles. It is concluded that single non-productive Dirofilaria worms elicit a strong expression of TGF-beta. This result is in accordance with observations on Onchocerca volvulus from patients with the hyporeactive (generalised) form.}, } @article {pmid20807539, year = {2010}, author = {Gavotte, L and Mercer, DR and Stoeckle, JJ and Dobson, SL}, title = {Costs and benefits of Wolbachia infection in immature Aedes albopictus depend upon sex and competition level.}, journal = {Journal of invertebrate pathology}, volume = {105}, number = {3}, pages = {341-346}, pmid = {20807539}, issn = {1096-0805}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; R01-AI067434/AI/NIAID NIH HHS/United States ; R01 AI067434-01A1/AI/NIAID NIH HHS/United States ; R01-AI051533/AI/NIAID NIH HHS/United States ; R01 AI051533-01A1/AI/NIAID NIH HHS/United States ; R01 AI067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Bacterial Infections/*physiopathology ; Female ; Male ; Pest Control, Biological/*methods ; Polymerase Chain Reaction ; Sex Ratio ; *Wolbachia ; }, abstract = {Bacterial endosymbionts induce various effects on hosts and can dramatically impact host fitness and development. An example is provided by obligate, maternally-inherited Wolbachia, which infect a broad range of invertebrates. Wolbachia are capable of altering host reproduction, thereby promoting infection spread. Wolbachia also pose direct physiological costs and benefits to hosts, complicating their categorization as parasites or mutualists. This study examines for an effect of Wolbachia infection in intra-specific larval competition by Aedes albopictus mosquitoes, with the goal of examining for an impact of Wolbachia infection in mixed populations. Similar to prior work examining for an influence of Wolbachia infection on the fitness of A. albopictus in adults, the results presented here support the hypothesized impact of Wolbachia across all life stages, including immatures. The differential competitiveness of infected larvae detected in our experiments indicates that Wolbachia infected A. albopictus females are less competitive relative to uninfected females when competing under highly competitive conditions. In contrast, under low competitive pressures, infected females experience higher survivorship. Thus, Wolbachia infection shifts from parasitism to mutualism as a function of developmental conditions. Results are discussed in relation to the invasion and persistence of Wolbachia in A. albopictus populations. The results are important to the evolution of stable Wolbachia symbioses, including Wolbachia invasion of an uninfected population. The resulting infection dynamics that occur in an infected population are discussed.}, } @article {pmid20739055, year = {2010}, author = {Taylor, MJ and Hoerauf, A and Bockarie, M}, title = {Lymphatic filariasis and onchocerciasis.}, journal = {Lancet (London, England)}, volume = {376}, number = {9747}, pages = {1175-1185}, doi = {10.1016/S0140-6736(10)60586-7}, pmid = {20739055}, issn = {1474-547X}, mesh = {Africa South of the Sahara ; Age Factors ; Albendazole/therapeutic use ; Animals ; Anti-Bacterial Agents/*therapeutic use ; Antinematodal Agents/*therapeutic use ; Blindness/parasitology ; Culicidae ; Dermatitis/parasitology ; Dermatologic Agents/therapeutic use ; Diethylcarbamazine/therapeutic use ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; *Elephantiasis, Filarial/complications/diagnosis/drug therapy/epidemiology/physiopathology/transmission ; Filaricides/therapeutic use ; Gram-Negative Bacterial Infections/complications ; Granuloma/parasitology ; Humans ; India ; Ivermectin/therapeutic use ; Lymphadenitis/parasitology ; Lymphangitis/parasitology ; Lymphedema/parasitology ; Macrolides/therapeutic use ; *Onchocerciasis/complications/diagnosis/drug therapy/epidemiology/physiopathology/transmission ; Prevalence ; Symbiosis ; Wolbachia/drug effects ; }, abstract = {Lymphatic filariasis and onchocerciasis are parasitic helminth diseases that constitute a serious public health issue in tropical regions. The filarial nematodes that cause these diseases are transmitted by blood-feeding insects and produce chronic and long-term infection through suppression of host immunity. Disease pathogenesis is linked to host inflammation invoked by the death of the parasite, causing hydrocoele, lymphoedema, and elephantiasis in lymphatic filariasis, and skin disease and blindness in onchocerciasis. Most filarial species that infect people co-exist in mutualistic symbiosis with Wolbachia bacteria, which are essential for growth, development, and survival of their nematode hosts. These endosymbionts contribute to inflammatory disease pathogenesis and are a target for doxycycline therapy, which delivers macrofilaricidal activity, improves pathological outcomes, and is effective as monotherapy. Drugs to treat filariasis include diethylcarbamazine, ivermectin, and albendazole, which are used mostly in combination to reduce microfilariae in blood (lymphatic filariasis) and skin (onchocerciasis). Global programmes for control and elimination have been developed to provide sustained delivery of drugs to affected communities to interrupt transmission of disease and ultimately eliminate this burden on public health.}, } @article {pmid20738784, year = {2010}, author = {Frost, CL and Fernández-Marín, H and Smith, JE and Hughes, WO}, title = {Multiple gains and losses of Wolbachia symbionts across a tribe of fungus-growing ants.}, journal = {Molecular ecology}, volume = {19}, number = {18}, pages = {4077-4085}, doi = {10.1111/j.1365-294X.2010.04764.x}, pmid = {20738784}, issn = {1365-294X}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Ants/*microbiology ; DNA, Bacterial/genetics ; Multilocus Sequence Typing ; *Phylogeny ; Species Specificity ; Symbiosis ; Wolbachia/classification/*genetics ; }, abstract = {Although the intracellular bacterium Wolbachia is ubiquitous in insects, it has a unique relationship with New World ants on which particular bacterial strains have specialized. However, data are from distantly related hosts and detailed phylogenetic information which could reveal transmission dynamics are lacking. Here, we investigate host-Wolbachia relationships in the monophyletic fungus-growing ant tribe Attini, screening 23 species and using multilocus sequence typing to reliably identify Wolbachia strains. This technique reduces the significant problem of recombination seen using traditional single gene techniques. The relationship between Wolbachia and the fungus-growing ants appears complex and dynamic. There is evidence of co-cladogenesis, supporting vertical transmission; however, this is incomplete, demonstrating that horizontal transmission has also occurred. Importantly, the infection prevalence is frequently different between closely related taxa, with the Acromyrmex leaf-cutting ants appearing particularly prone to infection and there being no consistent relationship with any of the major life history transitions. We suggest that infection loss and horizontal transmission have driven epidemics or selective sweeps of Wolbachia, resulting in multiple gains and losses of infection across the fungus-growing ants.}, } @article {pmid20730111, year = {2010}, author = {Slatko, BE and Taylor, MJ and Foster, JM}, title = {The Wolbachia endosymbiont as an anti-filarial nematode target.}, journal = {Symbiosis (Philadelphia, Pa.)}, volume = {51}, number = {1}, pages = {55-65}, pmid = {20730111}, issn = {0334-5114}, abstract = {Human disease caused by parasitic filarial nematodes is a major cause of global morbidity. The parasites are transmitted by arthropod intermediate hosts and are responsible for lymphatic filariasis (elephantiasis) or onchocerciasis (river blindness). Within these filarial parasites are intracellular alpha-proteobacteria, Wolbachia, that were first observed almost 30 years ago. The obligate endosymbiont has been recognized as a target for anti-filarial nematode chemotherapy as evidenced by the loss of worm fertility and viability upon antibiotic treatment in an extensive series of human trials. While current treatments with doxycycline and rifampicin are not practical for widespread use due to the length of required treatments and contraindications, anti-Wolbachia targeting nevertheless appears a promising alternative for filariasis control in situations where current programmatic strategies fail or are unable to be delivered and it provides a superior efficacy for individual therapy. The mechanisms that underlie the symbiotic relationship between Wolbachia and its nematode hosts remain elusive. Comparative genomics, bioinfomatic and experimental analyses have identified a number of potential interactions, which may be drug targets. One candidate is de novo heme biosynthesis, due to its absence in the genome sequence of the host nematode, Brugia malayi, but presence in Wolbachia and its potential roles in worm biology. We describe this and several additional candidate targets, as well as our approaches for understanding the nature of the host-symbiont relationship.}, } @article {pmid20728536, year = {2010}, author = {Sugimoto, TN and Fujii, T and Kayukawa, T and Sakamoto, H and Ishikawa, Y}, title = {Expression of a doublesex homologue is altered in sexual mosaics of Ostrinia scapulalis moths infected with Wolbachia.}, journal = {Insect biochemistry and molecular biology}, volume = {40}, number = {12}, pages = {847-854}, doi = {10.1016/j.ibmb.2010.08.004}, pmid = {20728536}, issn = {1879-0240}, mesh = {Amino Acid Sequence ; Animals ; DNA-Binding Proteins/*genetics/metabolism ; Female ; Gene Expression Regulation ; Insect Proteins/*genetics/metabolism ; Male ; Molecular Sequence Data ; Mosaicism ; Moths/drug effects/*genetics/metabolism/*microbiology ; Reverse Transcriptase Polymerase Chain Reaction ; Sex Determination Processes ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/*physiology ; }, abstract = {A homologue of the sex-determining gene doublesex, Osdsx, was identified in the adzuki bean borer Ostrinia scapulalis. Three isoforms of the Osdsx transcript (Osdsx(M), Osdsx(FL) and Osdsx(FS)) differing in length were found. Osdsx(M) was specifically found in males, and contained an 852-bp ORF encoding a protein of 284 amino acids. Osdsx(FL) and Osdsx(FS) were found in females, and had the same 813-bp ORF encoding a protein of 271 amino acids. The Osdsx gene was inferred to have six exons and five introns. The variation in the transcript could be explained by the alternative splicing of Osdsx: Osdsx(M) was formed by the splicing of exons 1, 2, 5 and 6, Osdsx(FS) by the splicing of exons 1-4 and 6, and Osdsx(FL) by the splicing of exons 1-6. RT-PCR analysis indicated that Osdsx was transcribed in a sex-specific manner in all somatic tissues examined, regardless of developmental stage. In Wolbachia-induced sexual mosaics of O. scapulalis, which are genetically male, the female-specific isoform of Osdsx (Osdsx(FL)) was shown to be expressed in addition to the male-specific isoform (Osdsx(M)). This finding provides the first evidence that Wolbachia manipulates the sex of its host by interfering either with the sex-specific splicing of Osdsx itself or with another upstream sex determination process.}, } @article {pmid20723069, year = {2010}, author = {Gueguen, G and Vavre, F and Gnankine, O and Peterschmitt, M and Charif, D and Chiel, E and Gottlieb, Y and Ghanim, M and Zchori-Fein, E and Fleury, F}, title = {Endosymbiont metacommunities, mtDNA diversity and the evolution of the Bemisia tabaci (Hemiptera: Aleyrodidae) species complex.}, journal = {Molecular ecology}, volume = {19}, number = {19}, pages = {4365-4376}, doi = {10.1111/j.1365-294X.2010.04775.x}, pmid = {20723069}, issn = {1365-294X}, mesh = {Animals ; Bacteria/*classification/genetics ; *Biological Evolution ; DNA, Mitochondrial/genetics ; DNA, Ribosomal Spacer/genetics ; Hemiptera/*genetics/*microbiology ; Linkage Disequilibrium ; Molecular Sequence Data ; *Phylogeny ; *Symbiosis ; }, abstract = {Bemisia tabaci, an invasive pest that causes crop damage worldwide, is a highly differentiated species complex, divided into biotypes that have mainly been defined based on mitochondrial DNA sequences. Although endosymbionts can potentially induce population differentiation, specialization and indirect selection on mtDNA, studies have largely ignored these influential passengers in B. tabaci, despite as many as seven bacterial endosymbionts have been identified. Here, we investigate the composition of the whole bacterial community in worldwide populations of B. tabaci, together with host genetic differentiation, focusing on the invasive B and Q biotypes. Among 653 individuals studied, more than 95% of them harbour at least one secondary endosymbiont, and multiple infections are very common. In addition, sequence analyses reveal a very high diversity of facultative endosymbionts in B. tabaci, with some bacterial genus being represented by more than one strain. In the B and Q biotypes, nine different strains of bacteria have been identified. The mtDNA-based phylogeny of B. tabaci also reveals a very high nucleotide diversity that partitions the two ITS clades (B and Q) into six CO1 genetic groups. Each genetic group is in linkage disequilibrium with a specific combination of endosymbionts. All together, our results demonstrate the rapid dynamics of the bacterial endosymbiont-host associations at a small evolutionary scale, questioning the role of endosymbiotic communities in the evolution of the Bemisia tabaci species complex and strengthening the need to develop a metacommunity theory of inherited endosymbionts.}, } @article {pmid20722287, year = {2010}, author = {Serga, SV and Demidov, SV and Kozeretskaia, IA}, title = {[Infection with Wolbachia does not influence crossing over in Drosophila melanogaster].}, journal = {TSitologiia i genetika}, volume = {44}, number = {4}, pages = {55-60}, pmid = {20722287}, issn = {0564-3783}, mesh = {Animals ; *Crossing Over, Genetic ; Drosophila melanogaster/*genetics/*microbiology ; Female ; Genes, Insect ; Male ; Mutation ; *Symbiosis ; Wolbachia/*physiology ; X Chromosome/genetics ; }, abstract = {The influence of infection with endosymbiotic bacteria Wolbachia on crossingover in Drosophila melanogaster between the white and cut genes in the X chromosome was studied. Reciprocal crosses have been conducted between infected and non-infected fruit fly strains. The results showed no significant effect of Wolbachia infection on the crossingover rates in D. melanogaster between the white and cut genes in X-chromosome.}, } @article {pmid20700535, year = {2010}, author = {Glaser, RL and Meola, MA}, title = {The native Wolbachia endosymbionts of Drosophila melanogaster and Culex quinquefasciatus increase host resistance to West Nile virus infection.}, journal = {PloS one}, volume = {5}, number = {8}, pages = {e11977}, pmid = {20700535}, issn = {1932-6203}, support = {R21 AI076258/AI/NIAID NIH HHS/United States ; AI076258/AI/NIAID NIH HHS/United States ; }, mesh = {Animal Feed/virology ; Animals ; Chikungunya virus/physiology ; Culex/*microbiology/physiology ; Drosophila melanogaster/*microbiology/physiology ; Female ; *Immunity, Innate ; Insect Vectors/microbiology/physiology ; La Crosse virus/physiology ; Male ; Phenotype ; *Symbiosis ; West Nile Fever/*immunology/microbiology/transmission ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The bacterial endosymbiont Wolbachia pipientis has been shown to increase host resistance to viral infection in native Drosophila hosts and in the normally Wolbachia-free heterologous host Aedes aegypti when infected by Wolbachia from Drosophila melanogaster or Aedes albopictus. Wolbachia infection has not yet been demonstrated to increase viral resistance in a native Wolbachia-mosquito host system.

In this study, we investigated Wolbachia-induced resistance to West Nile virus (WNV; Flaviviridae) by measuring infection susceptibility in Wolbachia-infected and Wolbachia-free D. melanogaster and Culex quinquefasciatus, a natural mosquito vector of WNV. Wolbachia infection of D. melanogaster induces strong resistance to WNV infection. Wolbachia-infected flies had a 500-fold higher ID50 for WNV and produced 100,000-fold lower virus titers compared to flies lacking Wolbachia. The resistance phenotype was transmitted as a maternal, cytoplasmic factor and was fully reverted in flies cured of Wolbachia. Wolbachia infection had much less effect on the susceptibility of D. melanogaster to Chikungunya (Togaviridae) and La Crosse (Bunyaviridae) viruses. Wolbachia also induces resistance to WNV infection in Cx. quinquefasciatus. While Wolbachia had no effect on the overall rate of peroral infection by WNV, Wolbachia-infected mosquitoes produced lower virus titers and had 2 to 3-fold lower rates of virus transmission compared to mosquitoes lacking Wolbachia.

CONCLUSIONS/SIGNIFICANCE: This is the first demonstration that Wolbachia can increase resistance to arbovirus infection resulting in decreased virus transmission in a native Wolbachia-mosquito system. The results suggest that Wolbachia reduces vector competence in Cx. quinquefasciatus, and potentially in other Wolbachia-infected mosquito vectors.}, } @article {pmid22127168, year = {2010}, author = {Giordano, R and Weber, E and Waite, J and Bencivenga, N and Krogh, PH and Soto-Adames, F}, title = {Effect of a high dose of three antibiotics on the reproduction of a parthenogenetic strain of Folsomia candida (Isotomidae: Collembola).}, journal = {Environmental entomology}, volume = {39}, number = {4}, pages = {1170-1177}, doi = {10.1603/EN10027}, pmid = {22127168}, issn = {1938-2936}, mesh = {Ampicillin/administration & dosage/toxicity ; Animals ; Anti-Bacterial Agents/administration & dosage/*toxicity ; Arthropods/*drug effects/microbiology ; Body Size/*drug effects ; Female ; Oviparity/*drug effects ; Ovum/*drug effects ; Oxytetracycline/administration & dosage/toxicity ; Parthenogenesis ; Symbiosis ; Tylosin/administration & dosage/toxicity ; Wolbachia ; }, abstract = {Folsomia candida Willem (Isotomidae: Collembola) is an edaphic parthenogenetic species commonly used in ecotoxicity studies. We exposed F. candida to a high dose of three antibiotics, tylosin, ampicillin, and oxytetracycline, that target different bacterial groups. Possible toxic effects were assessed through egg production, hatching, and body size. All three antibiotics caused toxic effects. Treatment with oxytetracycline proved the most toxic. This group showed the smallest body size and lowest number of eggs laid, likely the result of a combination of antibiotic toxicity and avoidance of the antibiotic spiked food. Active toxin avoidance by F. candida in toxicological assays may play a role in minimizing their exposure to toxic compounds. Despite the administration of high doses of oxytetracycline, F. candida individuals remained infected with the intracellular bacteria Wolbachia indicating that this strain is resistant to this antibiotic or that the host or its gut flora had detoxified the compound. An increase in percent egg hatch with time was seen in the ampicillin and oxytetracycline treatments, indicating a possible accommodation of the host and/or gut-flora to these antibiotics.}, } @article {pmid21594185, year = {2010}, author = {Arthofer, W and N Avtzis, D and Riegler, M and Stauffer, C}, title = {Mitochondrial phylogenies in the light of pseudogenes and Wolbachia: re-assessment of a bark beetle dataset.}, journal = {ZooKeys}, volume = {}, number = {56}, pages = {269-280}, pmid = {21594185}, issn = {1313-2970}, support = {P 21147/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Phylogenetic studies based on mtDNA become increasingly questioned because of potential pitfalls due to mitochondrial pseudogenes and mitochondrial selective sweeps. While the inclusion of nuclear markers should preferentially be considered for future studies, there is no need to abandon mtDNA as long as tests for the known mtDNA artefacts are performed. In this study we presentadditionaldata and test previous phylogeographical studies of Pityogenes chalcographus. We did not detect nuclear copies (numts) of the previously used mitochondrial markers by performing a combined long range/nested PCR of the COI gene and by an in silico analysis of the COI sequence data. This confirms the robustness of our previous phylogenetic study of Pityogenes chalcographus. Results of an in-situ hybridization of Wolbachia in Pityogenes chalcographus confirm the presence of this endosysmbiont in this species. However, we did not detect a correlation between infection status, geographical region and mtDNA haplotypes. The hybridisation data also support a previous hypothesis that infections do not result from parasitoids or parasitic nematodes, insect surface or laboratory contaminations and are hence a true infection of Pityogenes chalcographus. We conclude that the deep structure found in mitochondrial populations of Pityogenes chalcographus indeed represents the evolutionary history of European populations.}, } @article {pmid23129882, year = {2009}, author = {Paily, KP and Hoti, SL and Das, PK}, title = {A review of the complexity of biology of lymphatic filarial parasites.}, journal = {Journal of parasitic diseases : official organ of the Indian Society for Parasitology}, volume = {33}, number = {1-2}, pages = {3-12}, pmid = {23129882}, issn = {0971-7196}, abstract = {There are about five more common, including Wuchereria bancrofti and Brugia malayi, and four less common filarial parasites infecting human. Genetic analysis of W. bancrofti populations in India showed that two strains of the species are prevalent in the country. The adult filarial parasites are tissue specific in the human host and their embryonic stage, called microfilariae (mf), are found in the blood or skin of the host, depending upon the species of the parasite. Three genetically determined physiological races exist in W. bancrofti and B. malayi, based on the microfilarial periodicity. They are the nocturnally periodic, nocturnally subperiodic and diurnally subperiodic forms. The susceptibility of a mosquito species to filarial infection depends on various factors, which could be genetic, physiological or physical. Survival analysis of Culex quinquefasciatus infected with W. bancrofti showed that the parasite load in the mosquito is a risk factor of vector survival. The extrinsic life cycle of the parasite is initiated when the mf are ingested by a mosquito vector during feeding on the host blood. On maturity, most of the infective L3 stage larvae migrate to the head and proboscis of the mosquito to get transmitted to the mammalian host during subsequent feeding. They develop to the adult L5 stage and the period of development and the longevity of the parasites varies according to the species of the nematode and the mammalian host. The rate of production of mf by the adult female was found to be stable at least for a period of five years. The life span of the mf has some influence on the dynamics of transmission of filariasis. Recent studies show that the endosymbiont, Wolbachia, plays an important role in the survival of filarial parasites. The possibility of in vitro and in vivo culture of filarial parasites is also reviewed.}, } @article {pmid23100759, year = {2009}, author = {Rajagopal, R}, title = {Beneficial interactions between insects and gut bacteria.}, journal = {Indian journal of microbiology}, volume = {49}, number = {2}, pages = {114-119}, pmid = {23100759}, issn = {0046-8991}, abstract = {Insects are amongst the most successful of animals, both in terms of diversity and in colonizing all ecological niches. Recent studies have highlighted the benefi ciary roles that bacteria play in the success and establishment of insects. By adopting techniques like 16S rRNA sequencing we are now in a position to understand the diversity of bacteria present in insect guts. It has been shown that some of these bacteria, like Wolbachia and Cardinium are involved in manipulating insect populations and distorting their sex ratio. Attempts have been made to culture these bacteria in insect cell lines, as they are recalcitrant to culture under normal microbiological conditions. The diversity of bacteria associated with insects and the functional role played by them in the insect is discussed below.}, } @article {pmid21564565, year = {2009}, author = {Duplouy, A and Vermenot, C and Davies, N and Roderick, G and Hurst, GD and Charlat, S}, title = {Assessing risks of Wolbachia DNA cross-specimen contamination following mass collection and ethanol storage.}, journal = {Molecular ecology resources}, volume = {9}, number = {1}, pages = {46-50}, doi = {10.1111/j.1755-0998.2008.02421.x}, pmid = {21564565}, issn = {1755-098X}, abstract = {Wolbachia and other intracellular bacteria that manipulate reproduction are widespread and can have major consequences on the ecology and evolution of their hosts. Several studies have attempted to assess the host range of these bacteria based on polymerase chain reaction assays on material preserved and collected using a variety of methods. While collecting in the field, mass storage in ethanol before sorting specimens in the laboratory is by far the easiest technique, and an integral component of Malaise trapping. This implicitly relies on the assumption that mass ethanol storage does not produce cross-contamination of Wolbachia DNA among specimens. Here we test this assumption. The absence of cross contamination between known positive and negative samples stored within a vial indicate there is no reason to believe collective storage of specimens creates artefactual increases in the incidence of Wolbachia or other intracellular bacteria.}, } @article {pmid21585936, year = {2008}, author = {Unckless, RL and Boelio, LM and Schifano, M and Dyer, KA}, title = {Isolation and characterization of 30 polymorphic microsatellite loci from the mycophagous fly Drosophila innubila.}, journal = {Molecular ecology resources}, volume = {8}, number = {4}, pages = {939-942}, doi = {10.1111/j.1755-0998.2008.02124.x}, pmid = {21585936}, issn = {1755-098X}, abstract = {Drosophila innubila is a mushroom-feeding member of the quinaria group, found in the woodlands and forests of the 'sky islands' in Arizona and New Mexico and extending south into central Mexico. Here, we describe and characterize 30 polymorphic microsatellite loci from D. innubila collected in the Chiricahua Mountains in Arizona. The number of alleles ranged from three to 21, and observed heterozygosity ranged from 0.0513 to 0.9737. Six loci were putatively X-linked, six departed from Hardy-Weinberg equilibrium, seven had evidence of null alleles, and six showed evidence of linkage disequilibrium. These markers will be useful for examining population structure of D. innubila and its association with male-killing Wolbachia.}, } @article {pmid21412357, year = {2008}, author = {Shanchez-Contreras, M and Vlisidou, I}, title = {The diversity of insect-bacteria interactions and its applications for disease control.}, journal = {Biotechnology & genetic engineering reviews}, volume = {25}, number = {}, pages = {203-243}, doi = {10.5661/bger-25-203}, pmid = {21412357}, issn = {0264-8725}, mesh = {Animals ; Bacteria/genetics/pathogenicity ; Genetic Engineering ; *Host-Pathogen Interactions/genetics/physiology ; Insect Control/*methods ; Insect Vectors/microbiology ; Insecta/genetics/*microbiology/physiology ; Symbiosis/genetics/physiology ; Virulence ; Wolbachia/genetics/pathogenicity/physiology ; }, abstract = {Prokaryotic microorganisms are widespread in all environments on Earth, establishing diverse interactions with many eukaryotic taxa, including insects. These associations may be symbiotic, pathogenic and vectoring. Independently of the type of interaction, each association starts with the adhesion of the microorganism to the host, entry and "invasion" of the host, then progresses to establishment and dissemination within the host, by avoiding host immune responses, and concludes with transmission back to the environment or to a new host. Advances in genomics and genetics have allowed the dissection of these processes and provided important information on the elements driving the shaping of the members of each association. Furthermore, many mechanisms involved in the establishment of the associations have been scrutinised, along with the development of new methods for the management of insect populations.}, } @article {pmid24937885, year = {2004}, author = {Klei, TR and Chirgwin, SR and Solano-Aguilar, G and Shea-Donohue, T and Madden, K and Dawson, H and Ledbetter, T and Urban, J and Gasbarre, LC and Sonstegard, T and Van Tassell, CP and Araujo, R}, title = {AAVP Symposium: new approaches in the study of animal parasites.}, journal = {Veterinary parasitology}, volume = {125}, number = {1-2}, pages = {147-161}, doi = {10.1016/j.vetpar.2004.05.011}, pmid = {24937885}, issn = {1873-2550}, mesh = {Animals ; Genomics/trends ; Parasites/microbiology ; Parasitic Diseases, Animal/*parasitology ; Research/*trends ; Veterinary Medicine/*trends ; Wolbachia/physiology ; }, abstract = {The following three papers are a very small window onto the types of research being pursued by members of the American Association of Veterinary Parasitologists. They are related by the fact that newer areas in the biology of parasites and their hosts are discussed. The first paper by Dr. Tom Klei, gives a brief view of the interactions between host and parasite of the fascinating organism Wolbachia, a parasite of parasites. The second paper by Dr. Gloria Solano-Aguilar addresses the use of probiotics to alter the host–parasite interface and influence host resistance. The final paper by Dr. Lou Gasbarre outlines an example of integration of the genomics revolution into Veterinary Parasitology. While the subjects are diverse, they demonstrate the vitality of the AAVP.}, } @article {pmid24937882, year = {2004}, author = {, and Guerrero, J and McCall, JW and Genchi, C and Bazzocchi, C and Kramer, L and Simòn, F and Martarino, M}, title = {Recent advances in heartworm disease.}, journal = {Veterinary parasitology}, volume = {125}, number = {1-2}, pages = {105-130}, doi = {10.1016/j.vetpar.2004.05.008}, pmid = {24937882}, issn = {1873-2550}, mesh = {Animals ; Anthelmintics/*therapeutic use ; Dirofilariasis/*drug therapy ; Dog Diseases/*drug therapy ; Dogs ; Nematoda/physiology ; }, abstract = {This compilation of articles consists of four papers presented at the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP) (held in New Orleans, LA, USA, on 10–14 August 2003) in a symposium session titled “ Recent Advances in Heartworm Disease,” organized and chaired by JohnW. McCall and Jorge Guerrero. The first paper(Guerrero) covered the American Heartworm Society’s most recent revision of their guidelines for the diagnosis, prevention, and management of heartworm infection in dogs, based on new research and clinical experience, particularly in the areas of heartworm chemoprophylaxis, adulticide therapy,and serologic testing and retesting. The entire updated 2003 “Guidelines” are presented herein.One paper (McCall) reviewed the “soft-kill” adulticidal and “safety-net” (reach-back, retroactive,clinical prophylactic) activity of prolonged dosing of prophylactic doses of macrocyclic lactones,concluding that ivermectin is the most effective in this way, milbemycin oxime is the least effective,and the activity of injectable moxidectin and selamectin lies between that of ivermectin and milbemycin oxime. The two remaining papers provided an overview of the discovery, rediscovery,phylogeny, and biological association between Wolbachia endosymbionts and filarial nematodes(Genchi and co-authors) and compelling evidence that Wolbachia may play a major role in the immunopathogenesis of filarial diseases of man and animals (Kramer and co-authors).}, } @article {pmid21708727, year = {2002}, author = {Telschow, A and Hammerstein, P and Werren, JH}, title = {Effects of wolbachia on genetic divergence between populations: mainland-island model.}, journal = {Integrative and comparative biology}, volume = {42}, number = {2}, pages = {340-351}, doi = {10.1093/icb/42.2.340}, pmid = {21708727}, issn = {1540-7063}, abstract = {Cytoplasmic incompatibility (CI) induced by intracellular bacteria is a possible mechanism for speciation. Growing empirical evidence suggests that bacteria of the group Wolbachia may indeed act as isolating factors in recent insect speciation. Wolbachia are cytoplasmically transmitted and can cause uni- or bidirectional CI. We present a mainland-island model to investigate how much impact Wolbachia can have on genetic divergence between populations. In the first scenario we assume that the island population has diverged at a selected locus and ask whether genetic divergence will be maintained after introduction of migration from the mainland. In the second we explore whether divergence will originate under migration. For simplicity, the host organisms are modeled as haploid sexuals. Simulations show that if each population is initially infected with a different strain of Wolbachia, then higher levels of divergence occur at the locally selected locus than in the absence of Wolbachia. A weaker effect is seen when there is only unidirectional CI caused by a single strain of Wolbachia on the island. CI increases divergence because it reduces effective migration between mainland and island. Migrants suffer from being confronted with the wrong CI system and this also applies to their matrilineal descendants. Moreover, there is a strong linkage disequilibrium between host genotype and infection state, which helps to maintain Wolbachia differences between the populations in the face of migration A sex bias in migration can either increase or decrease the effect of Wolbachia on divergence. Results support the view that Wolbachia has the potential for increasing divergence between populations and thus could enhance probabilities of speciation.}, } @article {pmid21708726, year = {2002}, author = {Clark, ME and Karr, TL}, title = {Distribution of wolbachia within Drosophila reproductive tissue: implications for the expression of cytoplasmic incompatibility.}, journal = {Integrative and comparative biology}, volume = {42}, number = {2}, pages = {332-339}, doi = {10.1093/icb/42.2.332}, pmid = {21708726}, issn = {1540-7063}, abstract = {A PCR based quantitative assay was used to determine Wolbachia infection levels in three different Drosophila strains. In addition, confocal microscopy was used to confirm and calibrate these results. Wolbachia infection levels ranged from 2,600 to 18,500 per egg. Single ovaries and testes from each of the three strains were also assayed using the calibrated quantitative PCR assay. A general correlation was found between bacterial levels in eggs and those found in ovaries and testis. These infection levels were consistent with the expression of cytoplasmic incompatibility (CI). In two strains of D. simulans, although the overall bacterial numbers were not significantly different, they exhibited different levels of CI. A direct correlation between the number of infected developing sperm cysts in these strains and CI levels was observed. This calibrated assay should provide a useful baseline for future comparative work, particularly between laboratories.}, } @article {pmid21238270, year = {1998}, author = {Wilkinson, T}, title = {Wolbachia come of age.}, journal = {Trends in ecology & evolution}, volume = {13}, number = {6}, pages = {213-214}, doi = {10.1016/s0169-5347(98)01386-x}, pmid = {21238270}, issn = {0169-5347}, } @article {pmid24173522, year = {1996}, author = {Dobson, S and Tanouye, M}, title = {The paternal sex ratio chromosome induces chromosome loss independently of Wolbachia in the wasp Nasonia vitripennis.}, journal = {Development genes and evolution}, volume = {206}, number = {3}, pages = {207-217}, doi = {10.1007/s004270050046}, pmid = {24173522}, issn = {0949-944X}, abstract = {The paternal sex ratio chromosome (PSR) is a paternally-inherited supernumerary chromosome found in some males of Nasonia vitripennis. PSR induces the loss of N. vitripennis's paternal autosomes in early fertilized embryos. Previous examinations have not directly addressed the complication of PSR's co-occurrence with Wolbachia. Wolbachia is the name assigned to a group of cytoplasmic bacteria which induce numerous reproductive alterations in their hosts. In Nasonia, Wolbachia cause cytoplasmic incompatibility (CI) which also results in paternal chromosome loss. Here we address the question of whether PSR's function (i.e. PSR's transmission and/or ability to induce chromosome loss) depends upon or interacts with Wolbachia. A strain of PSR males is artificially cleared of Wolbachia. Test crosses and cytological observations of this strain demonstrate that PSR's transmission and ability to induce chromosome loss is not dependent upon Wolbachia. Comparisons suggest an absence of interactions between PSR and Wolbachia when they co-occur. Fluorescent and confocal microscopy are used to examine and compare early embryos. Observations demonstrate that microtubule interactions with chromatin do not appear to cause the initial loss of the paternal chromosomes. Cytological observations presented here also differ from previous reports of PSR- and Wolbachia-induced chromosome loss.}, } @article {pmid20700141, year = {2011}, author = {Watanabe, M and Miura, K and Hunter, MS and Wajnberg, E}, title = {Superinfection of cytoplasmic incompatibility-inducing Wolbachia is not additive in Orius strigicollis (Hemiptera: Anthocoridae).}, journal = {Heredity}, volume = {106}, number = {4}, pages = {642-648}, pmid = {20700141}, issn = {1365-2540}, mesh = {Animals ; Cytoplasm/genetics/*microbiology/physiology ; Female ; Hemiptera/genetics/*microbiology/physiology ; Male ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Cytoplasmic incompatibility (CI) allows the intracellular, maternally inherited bacterial symbiont Wolbachia to invade arthropod host populations by inducing infertility in crosses between infected males and uninfected females. The general pattern is consistent with a model of sperm modification, rescued only by egg cytoplasm infected with the same strain of symbiont. The predacious flower bug Orius strigicollis is superinfected with two strains of Wolbachia, wOus1 and wOus2. Typically, superinfections of CI Wolbachia are additive in their effects; superinfected males are incompatible with uninfected and singly infected females. In this study, we created an uninfected line, and lines singly infected with wOus1 and wOus2 by antibiotic treatment. Then, all possible crosses were conducted among the four lines. The results indicated that while wOus2 induces high levels of CI, wOus1 induces very weak or no CI, but can rescue CI caused by wOus2 to a limited extent. Levels of incompatibility in crosses with superinfected males did not show the expected pattern. In particular, superinfected males caused extremely weak CI when mated with either singly infected or uninfected females. An analysis of symbiont densities showed that wOus1 densities were significantly higher than wOus2 densities in superinfected males, and wOus2 densities were lower, but not significantly, in superinfected relative to singly infected males. These data lend qualified support for the hypothesis that wOus1 interferes with the ability of wOus2 to cause CI by suppressing wOus2 densities. To our knowledge, this is the first clear case of non-additive CI in a natural superinfection.}, } @article {pmid20695281, year = {2010}, author = {Ball, TS and Ritchie, SR}, title = {Sampling biases of the BG-sentinel trap with respect to physiology, age, and body size of adult Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {47}, number = {4}, pages = {649-656}, doi = {10.1603/me09218}, pmid = {20695281}, issn = {0022-2585}, mesh = {Aging/*physiology ; Animals ; Body Size/*physiology ; Culicidae/*physiology ; Entomology/*instrumentation ; Female ; Male ; Population Surveillance ; Selection Bias ; }, abstract = {Currently, Aedes aegypti (L.) control strategies are being developed that involve manipulation of the vector at the adult stage (e.g., the use of the bacterial endosymbiont Wolbachia to shorten the life span of the vector population). These novel strategies demand adult sampling methods to measure changes in population size, structure (age, sex ratio), and, ultimately, the success of the program. Each sampling method presents certain biases. Once these biases are defined, methods used to estimate population size and structure can be calibrated accordingly, resulting in more accurate and complex estimates of the vector population. A series of mark-release-recapture experiments with adult Ae. aegypti were conducted in a large outdoor flight cage and an indoor setting in far north Queensland, Australia. The biases of the BG-Sentinel trap (BGS) were investigated across several categories, as follows: 1) mosquito age; 2) sex; 3) physiological status; and 4) body size. Biases were not detected across age groups or body sizes. A significant bias was detected across physiological groups: nulliparous females were recaptured at a significantly lower rate than all other groups except blood-fed parous females, which were also recaptured at a low rate by the BGS. Males were recaptured at a higher rate than all groups, but only a significant difference in recapture rates was observed between males and nulliparous females. Previous studies show that the BGS is a highly effective tool for Ae. aegypti surveillance. The BGS proves to be a reliable tool in Ae. aegypti surveillance with consistent sampling outcomes. The sampling bias of the BGS is measurable and can be used to generate more accurate estimates of the adult population and its attributes.}, } @article {pmid20689574, year = {2010}, author = {Landmann, F and Foster, JM and Slatko, B and Sullivan, W}, title = {Asymmetric Wolbachia segregation during early Brugia malayi embryogenesis determines its distribution in adult host tissues.}, journal = {PLoS neglected tropical diseases}, volume = {4}, number = {7}, pages = {e758}, pmid = {20689574}, issn = {1935-2735}, mesh = {Animal Structures/microbiology ; Animals ; Brugia malayi/*growth & development/*microbiology ; Female ; Fluorescent Antibody Technique ; Male ; Staining and Labeling/methods ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia are required for filarial nematode survival and fertility and contribute to the immune responses associated with human filarial diseases. Here we developed whole-mount immunofluorescence techniques to characterize Wolbachia somatic and germline transmission patterns and tissue distribution in Brugia malayi, a nematode responsible for lymphatic filariasis. In the initial embryonic divisions, Wolbachia segregate asymmetrically such that they occupy only a small subset of cells in the developing embryo, facilitating their concentration in the adult hypodermal chords and female germline. Wolbachia are not found in male reproductive tissues and the absence of Wolbachia from embryonic germline precursors in half of the embryos indicates Wolbachia loss from the male germline may occur in early embryogenesis. Wolbachia rely on fusion of hypodermal cells to populate adult chords. Finally, we detect Wolbachia in the secretory canal lumen suggesting living worms may release bacteria and/or their products into their host.}, } @article {pmid20689172, year = {2010}, author = {Salunke, BK and Salunkhe, RC and Patole, MS and Shouche, YS}, title = {Wolbachia and termite association: present status and future implications.}, journal = {Journal of biosciences}, volume = {35}, number = {2}, pages = {171-175}, pmid = {20689172}, issn = {0973-7138}, mesh = {Animals ; Isoptera/*microbiology ; Phylogeny ; *Symbiosis ; Wolbachia/classification/genetics/*metabolism ; }, } @article {pmid20688703, year = {2010}, author = {Chinnadurai, SK and Birkenheuer, AJ and Blanton, HL and Maggi, RG and Belfiore, N and Marr, HS and Breitschwerdt, EB and Stoskopf, MK}, title = {Prevalence of selected vector-borne organisms and identification of Bartonella species DNA in North American river otters (Lontra canadensis).}, journal = {Journal of wildlife diseases}, volume = {46}, number = {3}, pages = {947-950}, doi = {10.7589/0090-3558-46.3.947}, pmid = {20688703}, issn = {1943-3700}, mesh = {Animals ; Animals, Wild/microbiology ; Bartonella/*isolation & purification ; Bartonella Infections/epidemiology/transmission/*veterinary ; DNA, Bacterial/blood ; Female ; Male ; North Carolina/epidemiology ; Otters/*virology ; Polymerase Chain Reaction/veterinary ; Prevalence ; Rivers ; }, abstract = {Trapper-killed North American river otters (Lontra canadensis) in North Carolina, USA, were screened for multiple vector-borne bacteria known to be pathogenic to mammals. Blood was collected from 30 carcasses in 2006, from 35 in 2007, and from one live otter in 2008. Samples were screened using conventional polymerase chain reaction (PCR) tests for DNA from Bartonella spp., Ehrlichia spp., and spotted fever group Rickettsia spp. All samples were negative for Rickettsia spp. Twelve of 30 samples from 2006 produced amplicons using the assay designed to detect Ehrlichia spp., but sequencing revealed that the amplified DNA fragment was from a novel Wolbachia sp., thought to be an endosymbiote of a Dirofilaria sp. Between 2006 and 2007, DNA from a novel Bartonella sp. was detected in 19 of 65 animals (29%). Blood from one live otter captured in 2008 was found positive for this Bartonella sp. by both PCR and culture. The pathogenicity of this Bartonella species in river otters or other mammals is unknown.}, } @article {pmid20676599, year = {2010}, author = {Sicard, M and Chevalier, F and De Vlechouver, M and Bouchon, D and Grève, P and Braquart-Varnier, C}, title = {Variations of immune parameters in terrestrial isopods: a matter of gender, aging and Wolbachia.}, journal = {Die Naturwissenschaften}, volume = {97}, number = {9}, pages = {819-826}, pmid = {20676599}, issn = {1432-1904}, mesh = {Aging ; Animals ; Crosses, Genetic ; Female ; Feminization ; Hemocytes/cytology/microbiology ; Hemolymph/physiology ; Immunocompetence ; Isopoda/*genetics/growth & development/immunology/microbiology ; Male ; Phenotype ; Sex Characteristics ; Sex Chromosomes/genetics ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Ecological factors modulate animal immunocompetence and potentially shape the evolution of their immune systems. Not only environmental parameters impact on immunocompetence: Aging is one major cause of variability of immunocompetence between individuals, and sex-specific levels of immunocompetence have also been frequently described. Moreover, a growing core of data put in light that vertically transmitted symbionts can dramatically modulate the immunocompetence of their hosts. In this study, we addressed the influence of gender, age and the feminising endosymbiont Wolbachia (wVulC) on variations in haemocyte density, total PO activity and bacterial load in the haemolymph of the terrestrial isopod Armadillidium vulgare. This host-symbiont system is of particular interest to address this question since: (1) wVulC was previously shown as immunosuppressive in middle-aged females and (2) wVulC influences sex determination. We show that age, gender and Wolbachia modulate together immune parameters in A. vulgare. However, wVulC, which interacts with aging, appears to be the prominent factor interfering with both PO activity and haemocyte density. This interference with immune parameters is not the only aspect of wVulC virulence on its host, as reproduction and survival are also altered.}, } @article {pmid20673071, year = {2010}, author = {Chu, D and Liu, G and Wan, F and Tao, Y and Gill, RJ}, title = {Phylogenetic analysis and rapid identification of the whitefly, Bemisia afer, in China.}, journal = {Journal of insect science (Online)}, volume = {10}, number = {}, pages = {86}, pmid = {20673071}, issn = {1536-2442}, mesh = {Animals ; China ; Endangered Species ; Female ; Genetic Markers ; Hemiptera/anatomy & histology/*classification/microbiology/physiology ; Male ; *Phylogeny ; Wolbachia/physiology ; }, abstract = {The phylogenetic relationship between the whitefly Bemisia afer (Priesner & Hosny) (Hemiptera: Aleyrodidae) from China and other populations among the world were analyzed based on the mitochondrial cytochrome oxidase I (mtCOI) gene. Phylogenetic analysis of mtCOI sequences and those of reference B. afer sequences showed that the populations of the species could be separated into 5 clades (I-V). There were at least two clades of the species from China (IV and V). These data suggested that B. afer might be a species complex. The Chinese B. afer populations were most divergent with B. afer from the United Kingdom and African countries. The distance between the Chinese B. afer (IV and V) and clades I, II, and III is more than 32%, while the distance among clades I, II, III is lower than 7.7%. A new set of primers specific to B. afer was designed to amplify a region of approximately 400 bp to discriminate B. afer from other Bemisia species in China based on mtCOI sequences.}, } @article {pmid20667099, year = {2010}, author = {Stouthamer, R and Russell, JE and Vavre, F and Nunney, L}, title = {Intragenomic conflict in populations infected by Parthenogenesis Inducing Wolbachia ends with irreversible loss of sexual reproduction.}, journal = {BMC evolutionary biology}, volume = {10}, number = {}, pages = {229}, pmid = {20667099}, issn = {1471-2148}, mesh = {Animals ; *Biological Evolution ; Cell Nucleus/genetics ; Female ; Fertilization ; *Genome, Insect ; Male ; Models, Biological ; Mutation ; Ovum ; Parthenogenesis/*genetics ; Sex Ratio ; *Symbiosis ; Wasps/*genetics/microbiology ; *Wolbachia ; }, abstract = {BACKGROUND: The maternally inherited, bacterial symbiont, parthenogenesis inducing (PI) Wolbachia, causes females in some haplodiploid insects to produce daughters from both fertilized and unfertilized eggs. The symbionts, with their maternal inheritance, benefit from inducing the production of exclusively daughters, however the optimal sex ratio for the nuclear genome is more male-biased. Here we examine through models how an infection with PI-Wolbachia in a previously uninfected population leads to a genomic conflict between PI-Wolbachia and the nuclear genome. In most natural populations infected with PI-Wolbachia the infection has gone to fixation and sexual reproduction is impossible, specifically because the females have lost their ability to fertilize eggs, even when mated with functional males.

RESULTS: The PI Wolbachia infection by itself does not interfere with the fertilization process in infected eggs, fertilized infected eggs develop into biparental infected females. Because of the increasingly female-biased sex ratio in the population during a spreading PI-Wolbachia infection, sex allocation alleles in the host that cause the production of more sons are rapidly selected. In haplodiploid species a reduced fertilization rate leads to the production of more sons. Selection for the reduced fertilization rate leads to a spread of these alleles through both the infected and uninfected population, eventually resulting in the population becoming fixed for both the PI-Wolbachia infection and the reduced fertilization rate. Fertilization rate alleles that completely interfere with fertilization ("virginity alleles") will be selected over alleles that still allow for some fertilization. This drives the final resolution of the conflict: the irreversible loss of sexual reproduction and the complete dependence of the host on its symbiont.

CONCLUSIONS: This study shows that dependence among organisms can evolve rapidly due to the resolution of the conflicts between cytoplasmic and nuclear genes, and without requiring a mutualism between the partners.}, } @article {pmid20651929, year = {2010}, author = {Sonthayanon, P and Peacock, SJ and Chierakul, W and Wuthiekanun, V and Blacksell, SD and Holden, MT and Bentley, SD and Feil, EJ and Day, NP}, title = {High rates of homologous recombination in the mite endosymbiont and opportunistic human pathogen Orientia tsutsugamushi.}, journal = {PLoS neglected tropical diseases}, volume = {4}, number = {7}, pages = {e752}, pmid = {20651929}, issn = {1935-2735}, support = {/WT_/Wellcome Trust/United Kingdom ; 073135/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Bacterial Typing Techniques/methods ; DNA Fingerprinting/methods ; DNA, Bacterial/chemistry/genetics ; Humans ; Mites/microbiology ; Orientia tsutsugamushi/*genetics/isolation & purification ; *Polymorphism, Genetic ; *Recombination, Genetic ; Scrub Typhus/microbiology ; Sequence Analysis, DNA/methods ; }, abstract = {Orientia tsutsugamushi is an intracellular alpha-proteobacterium which resides in trombiculid mites, and is the causative agent of scrub typhus in East Asia. The genome sequence of this species has revealed an unprecedented number of repeat sequences, most notably of the genes encoding the conjugative properties of a type IV secretion system (T4SS). Although this observation is consistent with frequent intragenomic recombination, the extent of homologous recombination (gene conversion) in this species is unknown. To address this question, and to provide a protocol for the epidemiological surveillance of this important pathogen, we have developed a multilocus sequence typing (MLST) scheme based on 7 housekeeping genes (gpsA, mdh, nrdB, nuoF, ppdK, sucD, sucB). We applied this scheme to the two published genomes, and to DNA extracted from blood taken from 84 Thai scrub typhus patients, from 20 cultured Thai patient isolates, 1 Australian patient sample, and from 3 cultured type strains. These data demonstrated that the O. tsutsugamushi population was both highly diverse [Simpson's index (95% CI) = 0.95 (0.92-0.98)], and highly recombinogenic. These results are surprising given the intracellular life-style of this species, but are broadly consistent with results obtained for Wolbachia, which is an alpha-proteobacterial reproductive parasite of arthropods. We also compared the MLST data with ompA sequence data and noted low levels of consistency and much higher discrimination by MLST. Finally, twenty-five percent of patients in this study were simultaneously infected with multiple sequence types, suggesting multiple infection caused by either multiple mite bites, or multiple strains co-existing within individual mites.}, } @article {pmid20647151, year = {2010}, author = {Cook, PE and McGraw, EA}, title = {Wolbachia pipientis: an expanding bag of tricks to explore for disease control.}, journal = {Trends in parasitology}, volume = {26}, number = {8}, pages = {373-375}, doi = {10.1016/j.pt.2010.05.006}, pmid = {20647151}, issn = {1471-5007}, mesh = {*Aedes/immunology/microbiology/parasitology/virology ; Animals ; Chikungunya virus/*growth & development ; Dengue Virus/*growth & development ; Immune System/immunology ; Insect Vectors/immunology/microbiology/parasitology/virology ; *Pest Control, Biological ; Plasmodium gallinaceum/*growth & development ; Wolbachia/*immunology ; }, abstract = {Wolbachia pipientis are maternally inherited, endosymbiotic bacteria that are widespread among insects. Two recent studies have demonstrated that Wolbachia inhibits the ability of medically significant pathogens, including filarial nematodes, dengue virus and Plasmodium to form infections in the mosquito vector, Aedes aegypti. We highlight the added value of these traits for Wolbachia based biocontrol strategies and evaluate the evidence for the idea that the insect immune response is responsible for the pathogen inhibition.}, } @article {pmid20644622, year = {2010}, author = {McMeniman, CJ and O'Neill, SL}, title = {A virulent Wolbachia infection decreases the viability of the dengue vector Aedes aegypti during periods of embryonic quiescence.}, journal = {PLoS neglected tropical diseases}, volume = {4}, number = {7}, pages = {e748}, pmid = {20644622}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Drosophila melanogaster/microbiology ; Ovum/microbiology ; Survival Analysis ; Wolbachia/isolation & purification/*pathogenicity ; }, abstract = {A new approach for dengue control has been proposed that relies on life-shortening strains of the obligate intracellular bacterium Wolbachia pipientis to modify mosquito population age structure and reduce pathogen transmission. Previously we reported the stable transinfection of the major dengue vector Aedes aegypti with a life-shortening Wolbachia strain (wMelPop-CLA) from the vinegar fly Drosophila melanogaster. Here, we report a further characterization of the phenotypic effects of this virulent Wolbachia infection on several life-history traits of Ae. aegypti. Minor costs of wMelPop-CLA infection for pre-imaginal survivorship, development and adult size were found. However, we discovered that the wMelPop-CLA infection dramatically decreased the viability of desiccated Ae. aegypti eggs over time. Similarly, the reproductive fitness of wMelPop-CLA infected Ae. aegypti females declined with age. These results reveal a general pattern associated with wMelPop-CLA induced pathogenesis in this mosquito species, where host fitness costs increase during aging of both immature and adult life-history stages. In addition to influencing the invasion dynamics of this particular Wolbachia strain, we suggest that the negative impact of wMelPop-CLA on embryonic quiescence may have applied utility as a tool to reduce mosquito population size in regions with pronounced dry seasons or in regions that experience cool winters.}, } @article {pmid20631135, year = {2010}, author = {Gottlieb, Y and Zchori-Fein, E and Mozes-Daube, N and Kontsedalov, S and Skaljac, M and Brumin, M and Sobol, I and Czosnek, H and Vavre, F and Fleury, F and Ghanim, M}, title = {The transmission efficiency of tomato yellow leaf curl virus by the whitefly Bemisia tabaci is correlated with the presence of a specific symbiotic bacterium species.}, journal = {Journal of virology}, volume = {84}, number = {18}, pages = {9310-9317}, pmid = {20631135}, issn = {1098-5514}, mesh = {Animals ; Bacterial Proteins/metabolism ; Begomovirus/*isolation & purification ; Chaperonin 60/metabolism ; DNA, Bacterial/chemistry/genetics ; Disease Vectors ; Enterobacteriaceae/metabolism/*physiology ; Hemiptera/*microbiology/*virology ; Israel ; Molecular Sequence Data ; Plant Diseases/*virology ; Protein Binding ; Sequence Analysis, DNA ; *Symbiosis ; Two-Hybrid System Techniques ; Wolbachia/metabolism/*physiology ; }, abstract = {Tomato yellow leaf curl virus (TYLCV) (Geminiviridae: Begomovirus) is exclusively vectored by the whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). TYLCV transmission depends upon a 63-kDa GroEL protein produced by the vector's endosymbiotic bacteria. B. tabaci is a species complex comprising several genetically distinct biotypes that show different secondary-symbiont fauna. In Israel, the B biotype harbors Hamiltonella, and the Q biotype harbors Wolbachia and Arsenophonus. Both biotypes harbor Rickettsia and Portiera (the obligatory primary symbionts). The aim of this study was to determine which B. tabaci symbionts are involved in TYLCV transmission using B. tabaci populations collected in Israel. Virus transmission assays by B. tabaci showed that the B biotype efficiently transmits the virus, while the Q biotype scarcely transmits it. Yeast two-hybrid and protein pulldown assays showed that while the GroEL protein produced by Hamiltonella interacts with TYLCV coat protein, GroEL produced by Rickettsia and Portiera does not. To assess the role of Wolbachia and Arsenophonus GroEL proteins (GroELs), we used an immune capture PCR (IC-PCR) assay, employing in vivo- and in vitro-synthesized GroEL proteins from all symbionts and whitefly artificial feeding through membranes. Interaction between GroEL and TYLCV was found to occur in the B biotype, but not in the Q biotype. This assay further showed that release of virions protected by GroEL occurs adjacent to the primary salivary glands. Taken together, the GroEL protein produced by Hamiltonella (present in the B biotype, but absent in the Q biotype) facilitates TYLCV transmission. The other symbionts from both biotypes do not seem to be involved in transmission of this virus.}, } @article {pmid20628578, year = {2010}, author = {Majerus, TM and Majerus, ME}, title = {Intergenomic arms races: detection of a nuclear rescue gene of male-killing in a ladybird.}, journal = {PLoS pathogens}, volume = {6}, number = {7}, pages = {e1000987}, pmid = {20628578}, issn = {1553-7374}, mesh = {Animals ; Biological Evolution ; Coleoptera/*genetics/*microbiology ; Female ; Genes, Bacterial ; Male ; Sex Characteristics ; *Sex Ratio ; Symbiosis/*genetics ; *Wolbachia ; }, abstract = {Many species of arthropod are infected by deleterious inherited micro-organisms. Typically these micro-organisms are inherited maternally. Consequently, some, particularly bacteria of the genus Wolbachia, employ a variety of strategies that favour female over male hosts. These strategies include feminisation, induction of parthenogenesis and male-killing. These strategies result in female biased sex ratios in host populations, which lead to selection for host factors that promote male production. In addition, the intra-genomic conflict produced by the difference in transmission of these cytoplasmic endosymbionts and nuclear factors will impose a pressure favouring nuclear factors that suppress the effects of the symbiont. During investigations of the diversity of male-killing bacteria in ladybirds (Coccinellidae), unexpected patterns of vertical transmission of a newly discovered male-killing taxon were observed in the ladybird Cheilomenes sexmaculata. Initial analysis suggested that the expression of the bacterial male-killing trait varies according to the male(s) a female has mated with. By swapping males between females, a male influence on the expression of the male-killing trait was confirmed. Experiments were then performed to determine the nature of the interaction. These studies showed that a single dominant allele, which rescues male progeny of infected females from the pathological effect of the male-killer, exists in this species. The gene shows typical Mendelian autosomal inheritance and is expressed irrespective of the parent from which it is inherited. Presence of the rescue gene in either parent does not significantly affect the inheritance of the symbiont. We conclude that C. sexmaculata is host to a male-killing gamma-proteobacterium. Further, this beetle is polymorphic for a nuclear gene, the dominant allele of which rescues infected males from the pathogenic effects of the male-killing agent. These findings represent the first reported case of a nuclear suppressor of male-killing in a ladybird. They are considered in regard to sex ratio and intra-genomic conflict theories, and models of the evolutionary dynamics and distribution of inherited symbionts.}, } @article {pmid20616278, year = {2010}, author = {Jaenike, J and Unckless, R and Cockburn, SN and Boelio, LM and Perlman, SJ}, title = {Adaptation via symbiosis: recent spread of a Drosophila defensive symbiont.}, journal = {Science (New York, N.Y.)}, volume = {329}, number = {5988}, pages = {212-215}, doi = {10.1126/science.1188235}, pmid = {20616278}, issn = {1095-9203}, mesh = {*Adaptation, Physiological ; Animals ; DNA, Mitochondrial/genetics ; Drosophila/genetics/microbiology/parasitology/*physiology ; Female ; Fertility ; Haplotypes ; Host-Parasite Interactions ; Molecular Sequence Data ; Polymerase Chain Reaction ; Spiroplasma/isolation & purification/*physiology ; *Symbiosis ; Tylenchida/anatomy & histology/*physiology ; Wolbachia/isolation & purification/physiology ; }, abstract = {Recent studies have shown that some plants and animals harbor microbial symbionts that protect them against natural enemies. Here we demonstrate that a maternally transmitted bacterium, Spiroplasma, protects Drosophila neotestacea against the sterilizing effects of a parasitic nematode, both in the laboratory and the field. This nematode parasitizes D. neotestacea at high frequencies in natural populations, and, until recently, almost all infections resulted in complete sterility. Several lines of evidence suggest that Spiroplasma is spreading in North American populations of D. neotestacea and that a major adaptive change to a symbiont-based mode of defense is under way. These findings demonstrate the profound and potentially rapid effects of defensive symbionts, which are increasingly recognized as major players in the ecology of species interactions.}, } @article {pmid20606691, year = {2011}, author = {White, JA and Kelly, SE and Cockburn, SN and Perlman, SJ and Hunter, MS}, title = {Endosymbiont costs and benefits in a parasitoid infected with both Wolbachia and Cardinium.}, journal = {Heredity}, volume = {106}, number = {4}, pages = {585-591}, pmid = {20606691}, issn = {1365-2540}, support = {2 K12 GM00708-06/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteroidetes/*physiology ; Female ; *Host-Pathogen Interactions ; Male ; Reproduction ; *Symbiosis ; Wasps/genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Theory suggests that maternally inherited endosymbionts can promote their spread and persistence in host populations by enhancing the production of daughters by infected hosts, either by improving overall host fitness, or through reproductive manipulation. In the doubly infected parasitoid wasp Encarsia inaron, Wolbachia manipulates host reproduction through cytoplasmic incompatibility (CI), but Cardinium does not. We investigated the fitness costs and/or benefits of infection by each bacterium in differentially cured E. inaron as a potential explanation for persistence of Cardinium in this population. We introgressed lines infected with Wolbachia, Cardinium or both with the cured line to create a similar genetic background, and evaluated several parasitoid fitness parameters. We found that symbiont infection resulted in both fitness costs and benefits for E. inaron. The cost was lower initial egg load for all infected wasps. The benefit was increased survivorship, which in turn increased male production for wasps infected with only Cardinium. Female production was unaffected by symbiont infection; we therefore have not yet identified a causal fitness effect that can explain the persistence of Cardinium in the population. Interestingly, the Cardinium survivorship benefit was not evident when Wolbachia was also present in the host, and the reproduction of doubly infected individuals did not differ significantly from uninfected wasps. Therefore, the results of our study show that even when multiple infections seem to have no effect on a host, there may be a complex interaction of costs and benefits among symbionts.}, } @article {pmid20603211, year = {2010}, author = {Crainey, JL and Hurst, J and Wilson, MD and Hall, A and Post, RJ}, title = {Construction and characterisation of a BAC library made from field specimens of the onchocerciasis vector Simulium squamosum (Diptera: Simuliidae).}, journal = {Genomics}, volume = {96}, number = {4}, pages = {251-257}, doi = {10.1016/j.ygeno.2010.06.004}, pmid = {20603211}, issn = {1089-8646}, support = {G0600015/MRC_/Medical Research Council/United Kingdom ; 77615/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Base Sequence ; Chromosome Mapping ; Chromosomes, Artificial, Bacterial/*genetics ; Cloning, Molecular ; *Disease Vectors ; Genes, Insect/genetics ; *Genomic Library ; Humans ; Microsatellite Repeats/genetics ; Onchocerciasis/parasitology/*transmission ; Simuliidae/*genetics/growth & development ; Specimen Handling/methods ; Wolbachia/genetics ; }, abstract = {A Bacterial Artificial Chromosome (BAC) library was made from wild-caught Simulium squamosum, which is an important vector of human onchocerciasis. The library is composed of 12,288 BACs, with an average insert size of 128 kb, and is expected to contain ~1.54 GB of cloned DNA. Random BAC-end sequencing generated over 95 kb of DNA sequence data from which putative S. squamosum gene sequences and novel repetitive DNA families were identified, including DNA transposons, retrotransposons and simple sequence repeats (SSRs). The sequence survey also provided evidence of DNA of microbial origin, and dissection of sample blackflies indicated that some of those used to prepare the library were likely to be parasitized by the mermithid Isomermis lairdi. Hybridisations with a set of three independent blackfly single-copy genes and two Wolbachia genes suggest that the library provides around 13-fold coverage of the S. squamosum genome and about 12-fold coverage of its Wolbachia endosymbiont.}, } @article {pmid20601501, year = {2010}, author = {Fu, Y and Gavotte, L and Mercer, DR and Dobson, SL}, title = {Artificial triple Wolbachia infection in Aedes albopictus yields a new pattern of unidirectional cytoplasmic incompatibility.}, journal = {Applied and environmental microbiology}, volume = {76}, number = {17}, pages = {5887-5891}, pmid = {20601501}, issn = {1098-5336}, support = {R01 AI067434/AI/NIAID NIH HHS/United States ; AI-67434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*growth & development/*microbiology ; Animals ; Cytoplasm/*microbiology ; Drosophila/microbiology ; Fertility ; Reproduction ; Symbiosis ; Wolbachia/growth & development/metabolism/*physiology ; }, abstract = {Obligately intracellular Wolbachia bacteria infect numerous invertebrates and often manipulate host reproduction to facilitate the spread of infection. An example of reproductive manipulation is Wolbachia-induced cytoplasmic incompatibility (CI), which occurs commonly in insects. This CI has been the focus both of basic scientific studies of naturally occurring invasion events and of applied investigations on the use of Wolbachia as a vehicle to drive desired genotypes into insect populations ("gene drive" or "population replacement" strategies). The latter application requires an ability to generate artificial infections that cause a pattern of unidirectional incompatibility with the targeted host population. A suggested target of population replacement strategies is the mosquito Aedes albopictus (Asian tiger mosquito), an important invasive pest and disease vector. Aedes albopictus individuals are naturally "superinfected" with two Wolbachia types: wAlbA and wAlbB. Thus, generating a strain that is unidirectionally incompatible with field populations requires the introduction of an additional infection into the preexisting superinfection. Although prior reports demonstrate an ability to transfer Wolbachia infections to A. albopictus artificially, including both intra- and interspecific Wolbachia transfers, previous efforts have not generated a strain capable of invading natural populations. Here we describe the generation of a stable triple infection by introducing Wolbachia wRi from Drosophila simulans into a naturally superinfected A. albopictus strain. The triple-infected strain displays a pattern of unidirectional incompatibility with the naturally infected strain. This unidirectional CI, combined with a high fidelity of maternal inheritance and low fecundity effects, suggests that the artificial cytotype could serve as an appropriate vehicle for gene drive.}, } @article {pmid20597993, year = {2010}, author = {Rodriguero, MS and Confalonieri, VA and Guedes, JV and Lanteri, AA}, title = {Wolbachia infection in the tribe Naupactini (Coleoptera, Curculionidae): association between thelytokous parthenogenesis and infection status.}, journal = {Insect molecular biology}, volume = {19}, number = {5}, pages = {631-640}, doi = {10.1111/j.1365-2583.2010.01018.x}, pmid = {20597993}, issn = {1365-2583}, mesh = {Animals ; Female ; Multilocus Sequence Typing ; Parthenogenesis ; Polymerase Chain Reaction ; Symbiosis ; Weevils/*microbiology/*physiology ; Wolbachia/*classification/genetics/physiology ; }, abstract = {Several parthenogenetic species of broad-nosed weevils exist, some of them of economic importance because of their pest status. Screening of the maternally inherited Wolbachia bacterium in 29 weevils of the tribe Naupactini, using multilocus sequence typing allowed us to assess a significant correlation between asexuality and infection, and suggests an involvement of Wolbachia in the origin of this reproductive mode. The nine Wolbachia strains retrieved from the Naupactini belong to the B supergroup. Phylogenetic analysis of these strains, along with other 23 strains obtained from arthropods and nematodes, supports previous hypotheses that horizontal transfer of Wolbachia amongst species from unrelated taxa has been pervasive.}, } @article {pmid20597434, year = {2010}, author = {Martin, C and Gavotte, L}, title = {The bacteria Wolbachia in filariae, a biological Russian dolls' system: new trends in antifilarial treatments.}, journal = {Parasite (Paris, France)}, volume = {17}, number = {2}, pages = {79-89}, doi = {10.1051/parasite/2010172079}, pmid = {20597434}, issn = {1252-607X}, mesh = {Animals ; Anthelmintics/therapeutic use ; Arthropods/microbiology ; Filariasis/*drug therapy/*microbiology/transmission ; Filarioidea/genetics/microbiology ; Gram-Negative Bacteria/isolation & purification ; Humans ; Vertebrates/parasitology ; Wolbachia/*genetics ; Wuchereria bancrofti/microbiology ; }, abstract = {Filarial nematode species can host Wolbachia bacterial endosymbionts. To understand the symbiosis, a higher level of complexity should be considered, taking in account the tripartite association between Wolbachia, filariae and mammals. This overview article discusses the biology of Wolbachia in filariae, including their distribution and phylogeny, mechanisms of action, inflammatory consequences on mammal host and biological control implications for filariases. Potential directions for future research are also discussed.}, } @article {pmid20596533, year = {2010}, author = {Barr, KL and Hearne, LB and Briesacher, S and Clark, TL and Davis, GE}, title = {Microbial symbionts in insects influence down-regulation of defense genes in maize.}, journal = {PloS one}, volume = {5}, number = {6}, pages = {e11339}, pmid = {20596533}, issn = {1932-6203}, mesh = {Animals ; *Down-Regulation ; *Genes, Plant ; Insecta/*microbiology ; Reverse Transcriptase Polymerase Chain Reaction ; *Symbiosis ; Wolbachia/*isolation & purification ; Zea mays/*genetics/immunology ; }, abstract = {Diabrotica virgifera virgifera larvae are root-feeding insects and significant pests to maize in North America and Europe. Little is known regarding how plants respond to insect attack of roots, thus complicating the selection for plant defense targets. Diabrotica virgifera virgifera is the most successful species in its genus and is the only Diabrotica beetle harboring an almost species-wide Wolbachia infection. Diabrotica virgifera virgifera are infected with Wolbachia and the typical gut flora found in soil-living, phytophagous insects. Diabrotica virgifera virgifera larvae cannot be reared aseptically and thus, it is not possible to observe the response of maize to effects of insect gut flora or other transient microbes. Because Wolbachia are heritable, it is possible to investigate whether Wolbachia infection affects the regulation of maize defenses. To answer if the success of Diabrotica virgifera virgifera is the result of microbial infection, Diabrotica virgifera virgifera were treated with antibiotics to eliminate Wolbachia and a microarray experiment was performed. Direct comparisons made between the response of maize root tissue to the feeding of antibiotic treated and untreated Diabrotica virgifera virgifera show down-regulation of plant defenses in the untreated insects compared to the antibiotic treated and control treatments. Results were confirmed via QRT-PCR. Biological and behavioral assays indicate that microbes have integrated into Diabrotica virgifera virgifera physiology without inducing negative effects and that antibiotic treatment did not affect the behavior or biology of the insect. The expression data and suggest that the pressure of microbes, which are most likely Wolbachia, mediate the down-regulation of many maize defenses via their insect hosts. This is the first report of a potential link between a microbial symbiont of an insect and a silencing effect in the insect host plant. This is also the first expression profile for a plant attacked by a root-feeding insect.}, } @article {pmid20585501, year = {2010}, author = {Negri, I and Pellecchia, M and Grève, P and Daffonchio, D and Bandi, C and Alma, A}, title = {Sex and stripping: The key to the intimate relationship between Wolbachia and host?.}, journal = {Communicative & integrative biology}, volume = {3}, number = {2}, pages = {110-115}, pmid = {20585501}, issn = {1942-0889}, abstract = {Wolbachia pipientis is known to infect only arthropods and nematodes (mainly filarial worms). A unique feature shared by the two Phyla is the ability to replace the exoskeleton, a process known as ecdysis. This shared characteristic is thought to reflect a common ancestry. Arthropod moulting is induced by the steroid hormone 20-hydroxyecdysone (20E) and a role for ecdysteroids in nematode ecdysis has also been suggested. Removing Wolbachia from filarial worms impairs the host's development. From analyses of the genome of Wolbachia harbored by the filarial nematode Brugia malayi and that of its host, the bacterium may provide a source of heme, an essential component of cytochrome P450's that are necessary for steroid hormone biosynthetic pathways.In arthropods, Wolbachia is a reproductive manipulator, inducing various phenotypic effects that may be due to differences in host physiology, in particular, endocrine-related processes governing development and reproduction. Insect steroids have well-defined roles in the coordination of multiple developmental processes, and in adults they control important aspects of reproduction, including ovarian development, oogenesis, sexual behavior, and in some taxa vitellogenin biosynthesis.According to some authors ecdysteroids may also act as sex hormones. In insects sex differentiation is generally thought to be a strictly genetic process, in which each cell decides its own sexual fate based on its sex chromosome constitution, but, surprisingly, recent data demonstrate that in Drosophila sex determination is not cell-autonomous, as it happens in mammals. Thus the presence of signals coordinating the development of a gender-specific phenotype cannot be excluded.This could explain why Wolbachia interferes with insect reproduction; and also could explain why Wolbachia interferes with insect development.Thus, is "sex (=reproduction) and stripping (=ecdysis)" the key to the intimate relationship between Wolbachia and its host?}, } @article {pmid20549240, year = {2010}, author = {Junpee, A and Tencomnao, T and Sanprasert, V and Nuchprayoon, S}, title = {Association between Toll-like receptor 2 (TLR2) polymorphisms and asymptomatic bancroftian filariasis.}, journal = {Parasitology research}, volume = {107}, number = {4}, pages = {807-816}, pmid = {20549240}, issn = {1432-1955}, mesh = {5' Untranslated Regions ; Adolescent ; Adult ; Animals ; Female ; Filariasis/*genetics/immunology/*parasitology ; Gene Frequency ; Genes, Reporter ; *Genetic Predisposition to Disease ; Haplotypes ; Humans ; Linkage Disequilibrium ; Luciferases/genetics/metabolism ; Male ; Polymorphism, Restriction Fragment Length ; *Polymorphism, Single Nucleotide ; Sequence Deletion ; Thailand ; Toll-Like Receptor 2/*genetics/immunology ; Transcription, Genetic ; Wuchereria bancrofti/*immunology/pathogenicity ; Young Adult ; }, abstract = {Lymphatic filariasis is mainly caused by the filarial nematodes Wuchereria bancrofti and Brugia malayi. Wolbachia, intracellular symbiotic bacteria in filarial parasite, is known to induce immune response predominantly through Toll-like receptor 2 (TLR2). This study was performed to investigate the association between polymorphisms of the TLR2 gene and susceptibility to asymptomatic bancroftian filariasis. A total of 142 unrelated asymptomatic bancroftian filariasis patients and 151 endemic normal controls in Tak province, Thailand were recruited into this study. The -196 to -173 deletion (del) polymorphism in the 5' untranslated region was investigated by allele-specific polymerase chain reaction. Two single nucleotide polymorphisms, +597 T>C and +1350 T>C, in exon 3 were identified by polymerase chain reaction-restriction fragment length polymorphism analysis. Furthermore, we analyzed the functional difference between the TLR2 -196 to -173 del and wild-type (wt) alleles using the luciferase reporter assay. All three polymorphisms were associated with a higher risk of asymptomatic bancroftian filariasis and were in strong linkage disequilibrium with each other. The TLR2 haplotype -196 to -173del/+597C/+1350C was strongly associated with an increased risk of asymptomatic bancroftian filariasis. The TLR2 -196 to -173 del allele had a significantly lower transcriptional activity than wt allele. The results of our study indicate that TLR2 -196 to -173 del, +597 T>C and +1350 T>C polymorphisms are associated with asymptomatic bancroftian filariasis in Thailand. Our functional study also supports this finding with respect to differential TLR2 gene expression by -196 to -173 del polymorphism.}, } @article {pmid20543958, year = {2010}, author = {McNulty, SN and Foster, JM and Mitreva, M and Dunning Hotopp, JC and Martin, J and Fischer, K and Wu, B and Davis, PJ and Kumar, S and Brattig, NW and Slatko, BE and Weil, GJ and Fischer, PU}, title = {Endosymbiont DNA in endobacteria-free filarial nematodes indicates ancient horizontal genetic transfer.}, journal = {PloS one}, volume = {5}, number = {6}, pages = {e11029}, pmid = {20543958}, issn = {1932-6203}, support = {T32 AI007172/AI/NIAID NIH HHS/United States ; T32-AI007172/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA, Bacterial/*genetics ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Nematoda/*microbiology ; RNA, Messenger/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; *Symbiosis ; Wolbachia/*genetics/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia are among the most abundant symbiotic microbes on earth; they are present in about 66% of all insect species, some spiders, mites and crustaceans, and most filarial nematode species. Infected filarial nematodes, including many pathogens of medical and veterinary importance, depend on Wolbachia for proper development and survival. The mechanisms behind this interdependence are not understood. Interestingly, a minority of filarial species examined to date are naturally Wolbachia-free.

We used 454 pyrosequencing to survey the genomes of two distantly related Wolbachia-free filarial species, Acanthocheilonema viteae and Onchocerca flexuosa. This screen identified 49 Wolbachia-like DNA sequences in A. viteae and 114 in O. flexuosa. qRT-PCR reactions detected expression of 30 Wolbachia-like sequences in A. viteae and 56 in O. flexuosa. Approximately half of these appear to be transcribed from pseudogenes. In situ hybridization showed that two of these pseudogene transcripts were specifically expressed in developing embryos and testes of both species.

CONCLUSIONS/SIGNIFICANCE: These results strongly suggest that the last common ancestor of extant filarial nematodes was infected with Wolbachia and that this former endosymbiont contributed to their genome evolution. Horizontally transferred Wolbachia DNA may explain the ability of some filarial species to live and reproduce without the endosymbiont while other species cannot.}, } @article {pmid20533041, year = {2011}, author = {Yun, Y and Lei, C and Peng, Y and Liu, F and Chen, J and Chen, L}, title = {Wolbachia strains typing in different geographic population spider, Hylyphantes graminicola (Linyphiidae).}, journal = {Current microbiology}, volume = {62}, number = {1}, pages = {139-145}, pmid = {20533041}, issn = {1432-0991}, mesh = {Animals ; Arachnida/*microbiology ; Bacterial Typing Techniques ; China ; Genotype ; Geography ; Multilocus Sequence Typing ; Phylogeny ; Symbiosis ; Wolbachia/*classification/genetics/*isolation & purification/physiology ; }, abstract = {The Wolbachia endosymbiont of spiders has not been extensively examined. In order to investigate the distribution, evolutionary history, and reproductive phenotype of Wolbachia in spiders in China, we tested 11 geographic populations of Hylyphantes graminicola. Wolbachia infection has been detected in each population. 10 Wolbachia strains have been characterized by multilocus sequence typing (MLST). Phylogenetic analyses indicated that eight Wolbachia strains in H. graminicola belonged to supergroup B, and two belonged to supergroup A. No correlation existed between Wolbachia diversity and host's geographic distance. The significant correlation was observed between pairwise distance of H. graminicola COI and genetic divergence of associated Wolbachia strains. We also found that Wolbachia infection frequencies in hosts varied over geographic space.}, } @article {pmid20529071, year = {2010}, author = {Stahlhut, JK and Desjardins, CA and Clark, ME and Baldo, L and Russell, JA and Werren, JH and Jaenike, J}, title = {The mushroom habitat as an ecological arena for global exchange of Wolbachia.}, journal = {Molecular ecology}, volume = {19}, number = {9}, pages = {1940-1952}, doi = {10.1111/j.1365-294X.2010.04572.x}, pmid = {20529071}, issn = {1365-294X}, mesh = {Agaricales ; Animals ; Bacterial Typing Techniques ; DNA, Bacterial/genetics ; Diptera/*microbiology ; *Food Chain ; Geography ; Models, Genetic ; Phylogeny ; Sequence Analysis, DNA ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia infect a variety of arthropod and nematode hosts, but in arthropods, host phylogenetic relationships are usually poor predictors of strain similarity. This suggests that new infections are often established by horizontal transmission. To gain insight into the factors affecting the probability of horizontal transmission among host species, we ask how host phylogeny, geographical distribution and ecology affect patterns of Wolbachia strain similarity. We used multilocus sequence typing (MLST) to characterize Wolbachia strain similarity among dipteran hosts associated with fleshy mushrooms. Wolbachia Supergroup A was more common than Supergroup B in Diptera, and also more common in mycophagous than non-mycophagous Diptera. Within Supergroup A, host family within Diptera had no effect on strain similarity, and there was no tendency for Wolbachia strains from sympatric host species to be more similar to one another than to strains from hosts in different biogeographical realms. Supergroup A strains differed between mycophagous and non-mycophagous Diptera more than expected by chance, suggesting that ecological associations can facilitate horizontal transmission of Wolbachia within mycophagous fly communities. For Supergroup B, there were no significant associations between strain similarity and host phylogeny, biogeography, or ecology. We identified only two cases in which closely related hosts carried closely related Wolbachia strains, evidence that Wolbachia-host co-speciation or early introgression can occur but may not be a major contributor to overall strain diversity. Our results suggest that horizontal transmission of Wolbachia can be influenced by host ecology, thus leading to partial restriction of Wolbachia strains or strain groups to particular guilds of insects.}, } @article {pmid20515696, year = {2010}, author = {Chang, J and Masters, A and Avery, A and Werren, JH}, title = {A divergent Cardinium found in daddy long-legs (Arachnida: Opiliones).}, journal = {Journal of invertebrate pathology}, volume = {105}, number = {3}, pages = {220-227}, doi = {10.1016/j.jip.2010.05.017}, pmid = {20515696}, issn = {1096-0805}, mesh = {Animals ; Arachnida/*microbiology ; Base Sequence ; Incidence ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal/genetics ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Recent studies indicate that a newly described bacterial endosymbiont, Cardinium, is widespread in arthropods and induces different reproductive manipulations in hosts. In this study, we used a portion of the 16S rRNA gene of the Cardinium to screen 16 Opilionid species from the suborder Palptores. We found the incidence of Cardinium in these Opiliones was significantly higher than in other pooled arthropods (31.2% versus 7.2%, P=0.007). Phylogenetic analyses using maximum parsimony (MP) and Bayesian analysis revealed two distinct clades in Opiliones. One is a divergent monophyletic clade with strong support that has so far not been found in other arthropods, and a second one contains Cardinium both from Opiliones and other arthropods. There is not complete concordance of the Cardinium strains with host phylogeny, suggesting some horizontal movement of the bacteria among Opiliones. Although the divergence in the sequenced 16S rRNA region between the Cardinium infecting Opiliones and Cardinium from other arthropods is greater than among Cardinium found in other arthropods, all are monophyletic with respect to the outgroup bacteria (endosymbionts of Acanthamoeba). Based on high pairwise genetic distances, deep branch, and a distinct phylogenetic grouping, we conclude that some Opiliones harbor a newly discovered Cardinium clade.}, } @article {pmid20500529, year = {2010}, author = {Küchler, SM and Dettner, K and Kehl, S}, title = {Molecular characterization and localization of the obligate endosymbiotic bacterium in the birch catkin bug Kleidocerys resedae (Heteroptera: Lygaeidae, Ischnorhynchinae).}, journal = {FEMS microbiology ecology}, volume = {73}, number = {2}, pages = {408-418}, doi = {10.1111/j.1574-6941.2010.00890.x}, pmid = {20500529}, issn = {1574-6941}, mesh = {Animals ; DNA, Bacterial/genetics ; Gammaproteobacteria/*classification/genetics/isolation & purification ; Heteroptera/*microbiology ; Larva/microbiology ; *Phylogeny ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {In contrast to specific bacterial symbionts of many stinkbugs, which are harboured extracellularly in the lumina of midgut sacs or tubular outgrowths, the obligate endosymbiont of birch catkin bug Kleidocerys resedae (Heteroptera: Lygaeidae) resides in a red-coloured, raspberry-shaped mycetome, localized abdominally, close to the midgut section. Phylogenetic analysis, based on the 16S rRNA gene and the groEL (chaperonin) gene, showed that the bacteria belong to the gamma-subdivision of the Proteobacteria and revealed a phylogenetic relationship with bacterial endosymbionts of Wigglesworthia glossinidia, the primary symbiont of tse-tse fly Glossina brevipalpis. Furthermore, RFLP analysis and sequencing revealed that K. resedae was also infected by Alphaproteobacteria of the genera Wolbachia and Rickettsia. The distribution and transmission of Kleidocerys endosymbiont in adults and all nymph stages were studied using FISH. The detection of symbionts at the anterior poles of developing eggs indicated that endosymbionts are transmitted vertically to offspring. Ultrastructural examinations by electron microscopy revealed the packed accommodation of rod-shaped bacteria in the cytoplasm of mycetocytes. A new genus and species name, 'Candidatus Kleidoceria schneideri', is proposed for this newly characterized clade of symbiotic bacteria.}, } @article {pmid20482609, year = {2010}, author = {Kremer, N and Dedeine, F and Charif, D and Finet, C and Allemand, R and Vavre, F}, title = {Do variable compensatory mechanisms explain the polymorphism of the dependence phenotype in the Asobara tabida-wolbachia association?.}, journal = {Evolution; international journal of organic evolution}, volume = {64}, number = {10}, pages = {2969-2979}, doi = {10.1111/j.1558-5646.2010.01034.x}, pmid = {20482609}, issn = {1558-5646}, mesh = {Animals ; Apoptosis ; Female ; Gene Expression Regulation ; *Genetic Variation ; Homeostasis ; Host-Pathogen Interactions ; Hymenoptera/genetics/microbiology/*physiology ; Iron/metabolism ; Oogenesis/genetics/physiology ; Oxidative Stress ; Phenotype ; *Polymorphism, Genetic ; Symbiosis ; Wolbachia/genetics/pathogenicity/*physiology ; }, abstract = {Wolbachia are symbiotic intracellular bacteria, which are classified as reproductive parasites. Although generally facultative, Wolbachia is necessary for Asobara tabida (Hymenoptera), because aposymbiotic females do not produce any offspring. Interestingly, the ovarian phenotype of aposymbiotic females is variable: some females do not produce any eggs, whereas others do produce some eggs, but these are aborted. Here, we show that the ovarian phenotype of aposymbiotic females is highly polymorphic within populations, although dependence remains complete in both cases. We also identified some lines in which aposymbiotic females were able to produce a very few viable offspring, further extending the range of variation observed. These results suggest that various factors actively maintain polymorphism. We demonstrated that Wolbachia is necessary to trigger oogenetic processes, but that the ovarian phenotype was determined by the host only. Phenotypic variation was also correlated with the differential expression of genes controlling iron homeostasis and oxidative stress, which are potentially involved in the evolution of dependence. This suggests that variation in the ovarian phenotype could reflect selection for different levels of compensatory mechanisms in response to Wolbachia infection, and that polymorphism is maintained through selection on different antagonist traits influenced by oxidative stress.}, } @article {pmid20462456, year = {2010}, author = {Zhang, F and Guo, H and Zheng, H and Zhou, T and Zhou, Y and Wang, S and Fang, R and Qian, W and Chen, X}, title = {Massively parallel pyrosequencing-based transcriptome analyses of small brown planthopper (Laodelphax striatellus), a vector insect transmitting rice stripe virus (RSV).}, journal = {BMC genomics}, volume = {11}, number = {}, pages = {303}, pmid = {20462456}, issn = {1471-2164}, mesh = {Animals ; Contig Mapping ; Expressed Sequence Tags ; Gene Expression Profiling/*methods ; Genes, Insect/genetics ; Hemiptera/*genetics/metabolism/physiology/virology ; Insect Proteins/genetics/metabolism ; Insect Vectors/*genetics/metabolism/physiology/virology ; Microsatellite Repeats/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA/*methods ; Symbiosis ; *Tenuivirus ; Wolbachia ; }, abstract = {BACKGROUND: The small brown planthopper (Laodelphax striatellus) is an important agricultural pest that not only damages rice plants by sap-sucking, but also acts as a vector that transmits rice stripe virus (RSV), which can cause even more serious yield loss. Despite being a model organism for studying entomology, population biology, plant protection, molecular interactions among plants, viruses and insects, only a few genomic sequences are available for this species. To investigate its transcriptome and determine the differences between viruliferous and naïve L. striatellus, we employed 454-FLX high-throughput pyrosequencing to generate EST databases of this insect.

RESULTS: We obtained 201,281 and 218,681 high-quality reads from viruliferous and naïve L. striatellus, respectively, with an average read length as 230 bp. These reads were assembled into contigs and two EST databases were generated. When all reads were combined, 16,885 contigs and 24,607 singletons (a total of 41,492 unigenes) were obtained, which represents a transcriptome of the insect. BlastX search against the NCBI-NR database revealed that only 6,873 (16.6%) of these unigenes have significant matches. Comparison of the distribution of GO classification among viruliferous, naïve, and combined EST databases indicated that these libraries are broadly representative of the L. striatellus transcriptomes. Functionally diverse transcripts from RSV, endosymbiotic bacteria Wolbachia and yeast-like symbiotes were identified, which reflects the possible lifestyles of these microbial symbionts that live in the cells of the host insect. Comparative genomic analysis revealed that L. striatellus encodes similar innate immunity regulatory systems as other insects, such as RNA interference, JAK/STAT and partial Imd cascades, which might be involved in defense against viral infection. In addition, we determined the differences in gene expression between vector and naïve samples, which generated a list of candidate genes that are potentially involved in the symbiosis of L. striatellus and RSV.

CONCLUSIONS: To our knowledge, the present study is the first description of a genomic project for L. striatellus. The identification of transcripts from RSV, Wolbachia, yeast-like symbiotes and genes abundantly expressed in viruliferous insect, provided a starting-point for investigating the molecular basis of symbiosis among these organisms.}, } @article {pmid20462452, year = {2010}, author = {Skaljac, M and Zanic, K and Ban, SG and Kontsedalov, S and Ghanim, M}, title = {Co-infection and localization of secondary symbionts in two whitefly species.}, journal = {BMC microbiology}, volume = {10}, number = {}, pages = {142}, pmid = {20462452}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; Croatia ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Geography ; Hemiptera/*growth & development/*microbiology ; In Situ Hybridization, Fluorescence ; Polymerase Chain Reaction ; Population Density ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 23S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {BACKGROUND: Whiteflies are cosmopolitan phloem-feeding pests that cause serious damage to many crops worldwide due to direct feeding and vectoring of many plant viruses. The sweetpotato whitefly Bemisia tabaci (Gennadius) and the greenhouse whitefly Trialeurodes vaporariorum (Westwood) are two of the most widespread and damaging whitefly species. To complete their unbalanced diet, whiteflies harbor the obligatory bacterium Portiera aleyrodidarum. B. tabaci further harbors a diverse array of secondary symbionts, including Hamiltonella, Arsenophonus, Cardinium, Wolbachia, Rickettsia and Fritschea. T. vaporariorum is only known to harbor P. aleyrodidarum and Arsenophonus. We conducted a study to survey the distribution of whitefly species in Croatia, their infection status by secondary symbionts, and the spatial distribution of these symbionts in the developmental stages of the two whitefly species.

RESULTS: T. vaporariorum was found to be the predominant whitefly species across Croatia, while only the Q biotype of B. tabaci was found across the coastal part of the country. Arsenophonus and Hamiltonella were detected in collected T. vaporariorum populations, however, not all populations harbored both symbionts, and both symbionts showed 100% infection rate in some of the populations. Only the Q biotype of B. tabaci was found in the populations tested and they harbored Hamiltonella, Rickettsia, Wolbachia and Cardinium, while Arsenophonus and Fritschea were not detected in any B. tabaci populations. None of the detected symbionts appeared in all populations tested, and multiple infections were detected in some of the populations. All endosymbionts tested were localized inside the bacteriocyte in both species, but only Rickettsia and Cardinium in B. tabaci showed additional localization outside the bacteriocyte.

CONCLUSIONS: Our study revealed unique co-infection patterns by secondary symbionts in B. tabaci and T. vaporariorum. Co-sharing of the bacteriocyte by the primary and different secondary symbionts is maintained through vertical transmission via the egg, and is unique to whiteflies. This system provides opportunities to study interactions among symbionts that co-inhabit the same cell in the same host: these can be cooperative or antagonistic, may affect the symbiotic contents over time, and may also affect the host by competing with the primary symbiont for space and resources.}, } @article {pmid20457156, year = {2010}, author = {Rossi, MI and Aguiar-Alves, F and Santos, S and Paiva, J and Bendas, A and Fernandes, O and Labarthe, N}, title = {Detection of Wolbachia DNA in blood from dogs infected with Dirofilaria immitis.}, journal = {Experimental parasitology}, volume = {126}, number = {2}, pages = {270-272}, doi = {10.1016/j.exppara.2010.05.002}, pmid = {20457156}, issn = {1090-2449}, mesh = {Animals ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/*blood/chemistry ; Dirofilaria immitis/*microbiology ; Dirofilariasis/blood/*microbiology ; Dog Diseases/blood/*microbiology/parasitology ; Dogs ; Female ; Male ; Microfilariae/growth & development/microbiology ; Polymerase Chain Reaction/methods ; Sensitivity and Specificity ; Symbiosis ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Dirofilaria immitis is the causative agent of heartworm disease in canines and felines, and pulmonary dirofilariasis in man. It harbors a symbiotic intracellular bacterium from the genus Wolbachia that plays an important role in its biology and contributes to the inflammatory pathology of the heartworm. This endosymbiont is sensitive to the tetracycline family of antibiotics prompting its use in the treatment of filariasis. To track Wolbachia during treatment, primers were designed based on the FtsZ gene from Wolbachia. These primers amplify a single PCR product with the expected size from DNA samples derived from various species of worms that harbor Wolbachia (D. immitis, Brugia malayi and Brugia pahangy). The detection limit of Wolbachia DNA in the assay was 80 pg of D. immitis DNA. Furthermore, the primer set successfully amplified the expected PCR product using blood samples from dogs harboring the heartworm and circulating microfilariae.}, } @article {pmid20442735, year = {2011}, author = {Russell, JE and Stouthamer, R}, title = {The genetics and evolution of obligate reproductive parasitism in Trichogramma pretiosum infected with parthenogenesis-inducing Wolbachia.}, journal = {Heredity}, volume = {106}, number = {1}, pages = {58-67}, pmid = {20442735}, issn = {1365-2540}, mesh = {Animals ; Biological Evolution ; Female ; Fertilization ; Hymenoptera/*genetics/*microbiology/physiology ; Male ; Parthenogenesis/*genetics ; Sex Ratio ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Parthenogenesis-inducing (PI) Wolbachia belong to a class of intracellular symbionts that distort the offspring sex ratio of their hosts toward a female bias. In many PI Wolbachia-infected species sex ratio distortion has reached its ultimate expression-fixation of infection and all-female populations. This is only possible with thelytokous PI symbionts as they provide an alternative form of reproduction and remove the requirement for males and sexual reproduction. Many populations fixed for PI Wolbachia infection have lost the ability to reproduce sexually, even when cured of the infection. We examine one such population in the species Trichogramma pretiosum. Through a series of backcrossing experiments with an uninfected Trichogramma pretiosum population we were able to show that the genetic basis for the loss of female sexual function could be explained by a dominant nuclear effect. Male sexual function had not been completely lost, though some deterioration of male sexual function was also evident when males from the infected population (created through antibiotic curing of infected females) were mated to uninfected females. We discuss the dynamics of sex ratio selection in PI Wolbachia-infected populations and the evolution of non-fertilizing mutations.}, } @article {pmid20413911, year = {2010}, author = {Gayen, P and Maitra, S and Datta, S and Babu, SP}, title = {Evidence for Wolbachia symbiosis in microfilariae of Wuchereria bancrofti from West Bengal, India.}, journal = {Journal of biosciences}, volume = {35}, number = {1}, pages = {73-77}, pmid = {20413911}, issn = {0973-7138}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/*pharmacology ; Elephantiasis, Filarial/*parasitology ; Female ; Geography ; Humans ; India ; Male ; Microscopy, Electron, Transmission/methods ; Middle Aged ; Polymerase Chain Reaction/methods ; RNA, Ribosomal, 16S/metabolism ; *Symbiosis ; Wolbachia/*physiology ; Wuchereria bancrofti/*microbiology/physiology ; }, abstract = {Wolbachia are symbiotic endobacteria that infect the majority of filarial nematodes, including Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus. Recent studies have suggested that Wolbachia are necessary for the reproduction and survival of filarial nematodes and have highlighted the use of antibiotic therapy such as tetracycline/doxycycline as a novel method of treatment for infections caused by these organisms. Before such therapy is conceived and implemented on a large scale, it is necessary to assess the prevalence of the endosymbiont in W. bancrofti from different geographical locations. We present data from molecular and electron microscopic studies to provide evidence for Wolbachia symbiosis in W. bancrofti microfilariae collected from two districts (Bankura and Birbhum) of West Bengal, India.}, } @article {pmid20405054, year = {2010}, author = {Turner, JD and Tendongfor, N and Esum, M and Johnston, KL and Langley, RS and Ford, L and Faragher, B and Specht, S and Mand, S and Hoerauf, A and Enyong, P and Wanji, S and Taylor, MJ}, title = {Macrofilaricidal activity after doxycycline only treatment of Onchocerca volvulus in an area of Loa loa co-endemicity: a randomized controlled trial.}, journal = {PLoS neglected tropical diseases}, volume = {4}, number = {4}, pages = {e660}, pmid = {20405054}, issn = {1935-2735}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Adolescent ; Adult ; Animals ; Double-Blind Method ; Doxycycline/*administration & dosage/adverse effects ; Endemic Diseases ; Female ; Filaricides/*administration & dosage/adverse effects ; Humans ; Ivermectin/administration & dosage/adverse effects ; Loiasis/complications/drug therapy/epidemiology ; Male ; Middle Aged ; Onchocerca volvulus/*drug effects ; Onchocerciasis/complications/*drug therapy ; Placebos/administration & dosage ; Treatment Outcome ; Young Adult ; }, abstract = {BACKGROUND: The risk of severe adverse events following treatment of onchocerciasis with ivermectin in areas co-endemic with loiasis currently compromises the development of control programmes and the treatment of co-infected individuals. We therefore assessed whether doxycycline treatment could be used without subsequent ivermectin administration to effectively deliver sustained effects on Onchocerca volvulus microfilaridermia and adult viability. Furthermore we assessed the safety of doxycycline treatment prior to ivermectin administration in a subset of onchocerciasis individuals co-infected with low to moderate intensities of Loa loa microfilaraemia.

METHODS: A double-blind, randomized, field trial was conducted of 6 weeks of doxycycline (200 mg/day) alone, doxycycline in combination with ivermectin (150 microg/kg) at +4 months or placebo matching doxycycline + ivermectin at +4 months in 150 individuals infected with Onchocerca volvulus. A further 22 individuals infected with O. volvulus and low to moderate intensities of Loa loa infection were administered with a course of 6 weeks doxycycline with ivermectin at +4 months. Treatment efficacy was determined at 4, 12 and 21 months after the start of doxycycline treatment together with the frequency and severity of adverse events.

RESULTS: One hundred and four (60.5%) participants completed all treatment allocations and follow up assessments over the 21-month trial period. At 12 months, doxycycline/ivermectin treated individuals had lower levels of microfilaridermia and higher frequency of amicrofilaridermia compared with ivermectin or doxycycline only groups. At 21 months, microfilaridermia in doxycycline/ivermectin and doxycycline only groups was significantly reduced compared to the ivermectin only group. 89% of the doxycycline/ivermectin group and 67% of the doxycycline only group were amicrofilaridermic, compared with 21% in the ivermectin only group. O. volvulus from doxycycline groups were depleted of Wolbachia and all embryonic stages in utero. Notably, the viability of female adult worms was significantly reduced in doxycycline treated groups and the macrofilaricidal and sterilising activity was unaffected by the addition of ivermectin. Treatment with doxycycline was well tolerated and the incidence of adverse event to doxycycline or ivermectin did not significantly deviate between treatment groups.

CONCLUSIONS: A six-week course of doxycycline delivers macrofilaricidal and sterilizing activities, which is not dependent upon co-administration of ivermectin. Doxycycline is well tolerated in patients co-infected with moderate intensities of L. loa microfilariae. Therefore, further trials are warranted to assess the safety and efficacy of doxycycline-based interventions to treat onchocerciasis in individuals at risk of serious adverse reactions to standard treatments due to the co-occurrence of high intensities of L. loa parasitaemias. The development of an anti-wolbachial treatment regime compatible with MDA control programmes could offer an alternative to the control of onchocerciasis in areas of co-endemicity with loiasis and at risk of severe adverse reactions to ivermectin.

TRIAL REGISTRATION: Controlled-Trials.com ISRCTN48118452.}, } @article {pmid20402779, year = {2010}, author = {Salunke, BK and Salunkhe, RC and Dhotre, DP and Khandagale, AB and Walujkar, SA and Kirwale, GS and Ghate, HV and Patole, MS and Shouche, YS}, title = {Diversity of Wolbachia in Odontotermes spp. (Termitidae) and Coptotermes heimi (Rhinotermitidae) using the multigene approach.}, journal = {FEMS microbiology letters}, volume = {307}, number = {1}, pages = {55-64}, doi = {10.1111/j.1574-6968.2010.01960.x}, pmid = {20402779}, issn = {1574-6968}, mesh = {Animals ; Bacterial Proteins/genetics ; *Biodiversity ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; India ; Isoptera/*microbiology ; Phylogeny ; Polymorphism, Genetic ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/*classification/*genetics ; }, abstract = {The intracellular bacteria, Wolbachia, are well known for inducing reproductive alterations in arthropod hosts, especially insects. The ancient origin and huge diversity, combined with the ecological, biological and behavioral plasticity of termites, make the latter exciting candidates for studying the interactions of Wolbachia. In the present study, we investigated the distribution of Wolbachia in populations of Odontotermes spp. and Coptotermes heimi termites occurring in 14 colonies (12 Odontotermes spp. and two C. heimi) from different locations in India. A striking diversity was observed among Wolbachia strains in closely related hosts based on five MLST genes (ftsZ, coxA, fbpA, hcpA and gatB) and the 16S rRNA gene. Wolbachia variants from two supergroups (B and F) were found in both the termite genera under study. This is the first report of Wolbachia infection in the Odontotermes genus. Although F Wolbachia supergroup infection is already reported in Coptotermes lacteus and Coptotermes acinaciformis, in this study, the two C. heimi species exhibited infection by two distinctly different Wolbachia supergroups (B and F).}, } @article {pmid20394655, year = {2010}, author = {Tien, NS and Sabelis, MW and Egas, M}, title = {The maintenance of genetic variation for oviposition rate in two-spotted spider mites: inferences from artificial selection.}, journal = {Evolution; international journal of organic evolution}, volume = {64}, number = {9}, pages = {2547-2557}, doi = {10.1111/j.1558-5646.2010.00996.x}, pmid = {20394655}, issn = {1558-5646}, mesh = {Animals ; Bacteroidetes/isolation & purification ; Crosses, Genetic ; Cytoplasm/microbiology ; Female ; Gene Frequency ; Male ; *Mutation ; Oviposition/*genetics ; Tetranychidae/*genetics/microbiology/physiology ; Wolbachia/isolation & purification ; }, abstract = {Despite the directional selection acting on life-history traits, substantial amounts of standing variation for these traits have frequently been found. This variation may result from balancing selection (e.g., through genetic trade-offs) or from mutation-selection balance. These mechanisms affect allele frequencies in different ways: Under balancing selection alleles are maintained at intermediate frequencies, whereas under mutation-selection balance variation is generated by deleterious mutations and removed by directional selection, which leads to asymmetry in the distribution of allele frequencies. To investigate the importance of these two mechanisms in maintaining heritable variation in oviposition rate of the two-spotted spider mite, we analyzed the response to artificial selection. In three replicate experiments, we selected for higher and lower oviposition rate, compared to control lines. A response to selection only occurred in the downward direction. Selection for lower oviposition rate did not lead to an increase in any other component of fitness, but led to a decline in female juvenile survival. The results suggest standing variation for oviposition rate in this population consists largely of deleterious alleles, as in a mutation-selection balance. Consequently, the standing variation for this trait does not appear to be indicative of its adaptive potential.}, } @article {pmid20388268, year = {2010}, author = {Zhang, X and Luckhart, S and Tu, Z and Pfeiffer, DG}, title = {Analysis of Wolbachia strains associated with Conotrachelus nenuphar (Coleoptera: Curculionidae) in the Eastern United States.}, journal = {Environmental entomology}, volume = {39}, number = {2}, pages = {396-405}, doi = {10.1603/EN09276}, pmid = {20388268}, issn = {1938-2936}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Female ; Male ; Phylogeny ; Prunus ; Symbiosis ; United States ; Weevils/*microbiology ; Wolbachia/*physiology ; }, abstract = {We studied the distribution patterns of Wolbachia infection associated with plum curculio strains in eight states of the eastern United States. The presence of the Wolbachia-specific gene wsp identified infections of this endosymbiont in 97.8% of the 93 samples tested. Three distinct Wolbachia strains were identified. The strains wCne1 (593 bp) and wCne2 (593 bp) were 97% identical, and their sequences were both 84% identical with wCne3 (590 bp). BLASTN searches through GenBank showed strong similarities between the wsp sequences of the three strains compared with Wolbachia sequenced from other hosts. Degree of similarity with sequences in other Wolbachia strains is discussed. Polymerase chain reaction-restriction fragment length polymorphism was used for superinfection detection. Of 93 samples, 15 (16.1%), 21 (22.6%), 19 (20.4%), and 36 (38.7%) samples were infected by wCne1, wCne2, wCne1 + 2, and wCne3, respectively. Only two (2.2%) samples had no infection. The wCne3 strain was always present as a single infection. Wolbachia strains approximate the distribution of plum curculio strains: northern strain infected with wCne1 and wCne2 strains in supergroup B, and southern strain infected with wCne3 strain in supergroup A, with the mid-Atlantic region as the convergence area. Based on haplotype distribution of plum curculio mitochondrial cytochrome oxidase I, there was a closer relation of the mid-southern plum curculio clade to the far-southern clade than to the northern clade. However, Wolbachia symbionts in mid-southern plum curculio are more closely related to those in northern plum curculio than to those in far-southern plum curculio. The relationship of Wolbachia infection with reproductive incompatibility between plum curculio populations was also discussed.}, } @article {pmid20386994, year = {2010}, author = {Psaila, R and Ponti, D and Ponzi, M and Gigliani, F and Battaglia, PA}, title = {A small sequence in domain v of the mitochondrial large ribosomal RNA restores Drosophila melanogaster pole cell determination in uv-irradiated embryos.}, journal = {Cellular & molecular biology letters}, volume = {15}, number = {3}, pages = {365-376}, pmid = {20386994}, issn = {1689-1392}, mesh = {Animals ; Base Sequence ; Cell Differentiation ; Cells, Cultured ; Drosophila melanogaster/*embryology/metabolism ; Embryo, Nonmammalian/*radiation effects ; Female ; Nucleic Acid Conformation ; Protein Binding ; RNA/*chemistry/metabolism ; RNA, Mitochondrial ; RNA, Ribosomal, 16S/*chemistry/metabolism ; RNA-Binding Proteins/metabolism ; *Ultraviolet Rays ; }, abstract = {The mechanism by which the mitochondrial large rRNA is involved in the restoration of the pole cell-forming ability in Drosophila embryos is still unknown. We identified a 15-ribonucleotide sequence which is conserved from the protobacterium Wolbachia to the higher eukaryotes in domain V of the mitochondrial large rRNA. This short sequence is sufficient to restore pole cell determination in UV-irradiated Drosophila embryos. Here, we provide evidence that the conserved 15-base sequence is sufficient to restore luciferase activity in vitro. Moreover, we show that the internal GAGA sequence is involved in protein binding and that mutations in this tetranucleotide affect the sequence's ability to restore luciferase activity. The obtained results lead us to propose that mtlrRNA may be involved either in damaged protein reactivation or in protein biosynthesis during pole cell determination.}, } @article {pmid20380298, year = {2010}, author = {Calvitti, M and Moretti, R and Lampazzi, E and Bellini, R and Dobson, SL}, title = {Characterization of a new Aedes albopictus (Diptera: Culicidae)-Wolbachia pipientis (Rickettsiales: Rickettsiaceae) symbiotic association generated by artificial transfer of the wPip strain from Culex pipiens (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {47}, number = {2}, pages = {179-187}, doi = {10.1603/me09140}, pmid = {20380298}, issn = {0022-2585}, mesh = {Aedes/*microbiology ; Animals ; Culex/*microbiology ; Female ; Longevity ; Male ; Reproduction ; *Symbiosis ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachia is a maternally inherited endosymbiont inducing various effects in insects and other invertebrate hosts that facilitate the invasion of naive host populations. One of the effects is a form of sterility known as cytoplasmic incompatibility (CI) through which females are effectively sterilized when they mate with males harboring a different Wolbachia strain. The repeated mass release of cytoplasmically incompatible males can be a tool to suppress insect populations. Here, we attempt to infect an Aedes albopictus (Skuse) (Diptera: Culicidae) strain, artificially deprived of the natural Wolbachia infection, with a new Wolbachia strain from Culex pipiens (L.) (Diptera: Culicidae). Further experiments were designed to study the effects of the new infection on Ae. albopictus fitness and evaluate key parameters that affect infection dynamics, including CI level and maternal inheritance. Using embryonic microinjection, the new Wolbachia strain was successfully established in Ae. albopictus. Crosses demonstrated a pattern of bidirectional CI between naturally infected and transinfected individuals. Specifically, egg hatch was essentially absent (i.e., CI was very high) in all crosses between the transinfected males and females with a different infection status. Furthermore, naturally infected Ae. albopictus males were incompatible with the transinfected females. Maternal inheritance was close to 100%. Moreover, the new infection did not affect immature and adult survivorship, but it significantly reduced female fecundity and egg hatch rate. The results are discussed in relation to the potential use of the new Ae. albopictus-Wolbachia symbiotic association as a suitable system for the study and development of CI-based strategies for suppressing populations of this important pest and disease vector.}, } @article {pmid20379652, year = {2010}, author = {de Almeida, RP and van Lenteren, JC and Stouthamer, R}, title = {Does Wolbachia infection affect Trichogramma atopovirilia behaviour?.}, journal = {Brazilian journal of biology = Revista brasleira de biologia}, volume = {70}, number = {2}, pages = {435-442}, doi = {10.1590/s1519-69842010005000016}, pmid = {20379652}, issn = {1678-4375}, mesh = {Animals ; Female ; Oviposition/*physiology ; Wasps/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Unisexual Trichogramma forms have attracted much attention due to their potential advantages as biocontrol agents. Fitness studies have been performed and understanding the cost that Wolbachia may inflict on their hosts will help in deciding if Wolbachia infected (unisexual) forms are indeed better than sexual forms when used in biological control programmes. The influence of Wolbachia on the foraging behaviour (including walking activity and speed) of T. atopovirilia is reported in this paper. Temperature strongly affected T. atopovirilia female walking activity, but Wolbachia infected and uninfected females differed in none of the behavioural components that were measured such as walking activity and walking speed. Walking activity was highest at 25 masculineC and differed significantly from that at 20 and 15 masculineC. Trichogramma wasps were highly affected at 15 masculineC. Behaviour analysis with females showed that female wasps spend most of the time on drilling + ovipositing on host eggs followed by host drumming and walking while drumming. The parasitism rate and number of offspring did not differ significantly between infected and cured Trichogramma females. Biological control implications of these findings are discussed.}, } @article {pmid20368968, year = {2010}, author = {Bian, G and Xu, Y and Lu, P and Xie, Y and Xi, Z}, title = {The endosymbiotic bacterium Wolbachia induces resistance to dengue virus in Aedes aegypti.}, journal = {PLoS pathogens}, volume = {6}, number = {4}, pages = {e1000833}, pmid = {20368968}, issn = {1553-7374}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; R01 AI-080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/immunology/*microbiology ; Animals ; Dengue/transmission ; Dengue Virus/*physiology ; Fluorescent Antibody Technique ; Host-Parasite Interactions/physiology ; Insect Vectors/immunology/*microbiology ; Pest Control, Biological/*methods ; Reverse Transcriptase Polymerase Chain Reaction ; Symbiosis/*physiology ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Genetic strategies that reduce or block pathogen transmission by mosquitoes have been proposed as a means of augmenting current control measures to reduce the growing burden of vector-borne diseases. The endosymbiotic bacterium Wolbachia has long been promoted as a potential vehicle for introducing disease-resistance genes into mosquitoes, thereby making them refractory to the human pathogens they transmit. Given the large overlap in tissue distribution and intracellular localization between Wolbachia and dengue virus in mosquitoes, we conducted experiments to characterize their interactions. Our results show that Wolbachia inhibits viral replication and dissemination in the main dengue vector, Aedes aegypti. Moreover, the virus transmission potential of Wolbachia-infected Ae. aegypti was significantly diminished when compared to wild-type mosquitoes that did not harbor Wolbachia. At 14 days post-infection, Wolbachia completely blocked dengue transmission in at least 37.5% of Ae. aegypti mosquitoes. We also observed that this Wolbachia-mediated viral interference was associated with an elevated basal immunity and increased longevity in the mosquitoes. These results underscore the potential usefulness of Wolbachia-based control strategies for population replacement.}, } @article {pmid20362581, year = {2010}, author = {Strübing, U and Lucius, R and Hoerauf, A and Pfarr, KM}, title = {Mitochondrial genes for heme-dependent respiratory chain complexes are up-regulated after depletion of Wolbachia from filarial nematodes.}, journal = {International journal for parasitology}, volume = {40}, number = {10}, pages = {1193-1202}, doi = {10.1016/j.ijpara.2010.03.004}, pmid = {20362581}, issn = {1879-0135}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; DNA, Complementary/genetics/metabolism ; Doxycycline/pharmacology ; Electron Transport/genetics/*physiology ; Filarioidea/genetics/*metabolism ; Genes, Mitochondrial/genetics/*physiology ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Up-Regulation ; Wolbachia/drug effects/*physiology ; }, abstract = {The filarial nematodes Brugia malayi, Wuchereria bancrofti and Onchocerca volvulus cause elephantiasis or dermatitis and blindness resulting in severe morbidity. Annually, 1.3 billion people are at risk of infection. Targeting the essential Wolbachia endobacteria of filarial nematodes with doxycycline has proven to be an effective therapy resulting in a block in embryogenesis, worm development and macrofilaricidal effects. However, doxycycline is contraindicated for a large portion of the at risk population. To identify new targets for anti-wolbachial therapy, understanding the molecular basis of the Wolbachia-filaria symbiosis is required. Using the B. malayi microarray we identified differentially expressed genes in the rodent filaria Litomosoides sigmodontis after depletion of Wolbachia which might have a role in symbiosis. The microarray data were filtered for regulated genes with a false discovery rate <5% and a > or = 2-fold-change. Most of the genes were differentially expressed at day 36 of tetracycline treatment, when 99.8% of Wolbachia were depleted. Several classes of genes were affected, including genes for translation, transcription, folding/sorting of proteins, motility, structure and metabolic and signalling pathways. Quantitative PCR validated 60% of the genes found to be regulated in the microarray. A nuclear encoded heme-binding protein of the globin family was up-regulated upon loss of Wolbachia. Interestingly, mitochondrial encoded subunits of respiratory chain complexes containing heme and riboflavin were also up-regulated. No change in the expression of these genes was seen in tetracycline treated Wolbachia-free Acanthocheilonema viteae. As Wolbachia synthesise heme and filaria do not, we hypothesise that without the endosymbionts no functional heme-containing enzymes can be formed, leading to loss of energy metabolism which then results in up-regulation of the mitochondrial encoded subunits in an attempt to correct the deviation from homeostasis. Our results support targeting the Wolbachia heme synthesis pathway for the discovery of new anti-filarial drugs.}, } @article {pmid20359544, year = {2010}, author = {Korten, S and Kaifi, JT and Büttner, DW and Hoerauf, A}, title = {Transforming growth factor-beta expression by host cells is elicited locally by the filarial nematode Onchocerca volvulus in hyporeactive patients independently from Wolbachia.}, journal = {Microbes and infection}, volume = {12}, number = {7}, pages = {555-564}, doi = {10.1016/j.micinf.2010.03.011}, pmid = {20359544}, issn = {1769-714X}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Antiparasitic Agents/therapeutic use ; Doxycycline/therapeutic use ; Endothelium/metabolism ; Female ; HIV Infections/complications/metabolism ; Host-Parasite Interactions ; Humans ; Ivermectin/therapeutic use ; Lymphocytes/metabolism ; Macrophages/metabolism ; Male ; Mast Cells/metabolism ; Onchocerca volvulus/*physiology ; Onchocerciasis/drug therapy/*metabolism/microbiology ; Rickettsiaceae Infections/complications ; Transforming Growth Factor beta/*metabolism ; *Wolbachia ; }, abstract = {Transforming growth factor-beta (TGF-beta) is a key cytokine in immune regulation, cell differentiation, development, wound healing, and tissue remodelling. It mediates immunosuppression in filarial infections facilitating parasite persistence, while attenuating immunopathology, which is induced by migrating microfilariae. Immunosuppression rises with parasite burden, but it remains unknown whether filariae elicit local release of immunosuppressive cytokines. Therefore, using immunohistology, we investigated the expression of stable, released latent TGF-beta1 in subcutaneous nodules from highly infected, hyporeactive onchocerciasis patients, harbouring adult Onchocerca volvulus. Since many cell types produce TGF-beta, we elucidated the cellular source, distribution and dependency on the worms' sex, productivity and vitality. We found TGF-beta1 to be abundantly expressed by T cells, plasma/B cells, macrophages, mast cells, fibrocytes, and vascular endothelial cells, particularly in onchocercomas with productive or previously productive females, damaged, dead and resorbed adult worms or microfilariae. We conclude TGF-beta to be antigen induced by the filariae since expression was scarce around subcutaneous arthropods or cholesterol crystals in onchocercomas. Enhanced expression after ivermectin or endobacteria-depleting doxycycline treatment indicates induction to depend on filariae and not on Wolbachia endobacteria. TGF-beta(+) cells were reduced in HIV co-infection. This finding of local and sustained TGF-beta induction by vital and dead filariae, untreated and after treatment, adds new aspects to immunomodulation by helminths.}, } @article {pmid20356892, year = {2010}, author = {Kaiser, W and Huguet, E and Casas, J and Commin, C and Giron, D}, title = {Plant green-island phenotype induced by leaf-miners is mediated by bacterial symbionts.}, journal = {Proceedings. Biological sciences}, volume = {277}, number = {1692}, pages = {2311-2319}, pmid = {20356892}, issn = {1471-2954}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Base Sequence ; Cytokinins/analysis ; DNA, Bacterial/chemistry/genetics ; Female ; Lepidoptera/*microbiology ; Male ; *Malus ; Molecular Sequence Data ; Plant Diseases/*parasitology ; Plant Leaves/*parasitology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/chemistry/genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Statistics, Nonparametric ; *Symbiosis ; Wolbachia/genetics/*growth & development ; }, abstract = {The life cycles of many organisms are constrained by the seasonality of resources. This is particularly true for leaf-mining herbivorous insects that use deciduous leaves to fuel growth and reproduction even beyond leaf fall. Our results suggest that an intimate association with bacterial endosymbionts might be their way of coping with nutritional constraints to ensure successful development in an otherwise senescent environment. We show that the phytophagous leaf-mining moth Phyllonorycter blancardella (Lepidoptera) relies on bacterial endosymbionts, most likely Wolbachia, to manipulate the physiology of its host plant resulting in the 'green-island' phenotype--photosynthetically active green patches in otherwise senescent leaves--and to increase its fitness. Curing leaf-miners of their symbiotic partner resulted in the absence of green-island formation on leaves, increased compensatory larval feeding and higher insect mortality. Our results suggest that bacteria impact green-island induction through manipulation of cytokinin levels. This is the first time, to our knowledge, that insect bacterial endosymbionts have been associated with plant physiology.}, } @article {pmid20345686, year = {2010}, author = {Mousson, L and Martin, E and Zouache, K and Madec, Y and Mavingui, P and Failloux, AB}, title = {Wolbachia modulates Chikungunya replication in Aedes albopictus.}, journal = {Molecular ecology}, volume = {19}, number = {9}, pages = {1953-1964}, doi = {10.1111/j.1365-294X.2010.04606.x}, pmid = {20345686}, issn = {1365-294X}, mesh = {Aedes/*microbiology/physiology/*virology ; Alphavirus Infections/*microbiology ; Animals ; Chikungunya virus/genetics/*physiology ; DNA, Bacterial/genetics ; Female ; Oviposition ; RNA, Viral/genetics ; Salivary Glands/virology ; Symbiosis ; *Virus Replication ; Wolbachia/genetics/*physiology ; }, abstract = {The Aedes albopictus mosquito has been involved as the principal vector of recent major outbreaks due to the chikungunya virus (CHIKV). The species is naturally infected by two strains of Wolbachia (wAlbA and wAlbB). Wolbachia infections are thought to have spread by manipulating the reproduction of their hosts; cytoplasmic incompatibility is the mechanism used by Wolbachia to invade natural populations of many insects including Ae. albopictus. Here, we report a study on the effects of removing Wolbachia from Ae. albopictus on CHIKV replication and examine the consequences of CHIKV infection on some life-history traits (survival and reproduction) of Wolbachia-free Ae. albopictus. We found that Wolbachia-free mosquitoes maintained a highly heterogeneous CHIKV replication compared to Wolbachia-infected individuals. In Wolbachia-infected Ae. albopictus, the regular increase of CHIKV followed by a steady viral load from day 4 post-infection onwards was concomitant with a decline in Wolbachia density. This profile was also detected when examining the two key organs for viral transmission, the midgut and the salivary glands. Moreover, Wolbachia-free Ae. albopictus was not altered in life-history traits such as survival, oviposition and hatching characteristics whether infected or not with CHIKV. We found that Wolbachia is not essential for viral replication, its presence could lead to optimize replication from day 4 post-infection onwards, coinciding with a decrease in Wolbachia density. Wolbachia may regulate viral replication in Ae. albopictus, with consequences on survival and reproduction.}, } @article {pmid20345674, year = {2010}, author = {Reumer, BM and VAN Alphen, JJ and Kraaijeveld, K}, title = {Ecology, Wolbachia infection frequency and mode of reproduction in the parasitoid wasp Tetrastichus coeruleus (Hymenoptera: Eulophidae).}, journal = {Molecular ecology}, volume = {19}, number = {8}, pages = {1733-1744}, doi = {10.1111/j.1365-294X.2010.04599.x}, pmid = {20345674}, issn = {1365-294X}, mesh = {Animals ; DNA, Bacterial/genetics ; Ecosystem ; Female ; France ; Male ; Netherlands ; *Parthenogenesis ; Sequence Analysis, DNA ; Sex Ratio ; United States ; Wasps/*microbiology/*physiology ; Wolbachia/*genetics ; }, abstract = {Whereas sexual reproduction may facilitate adaptation to complex environments with many biotic interactions, simplified environments are expected to favour asexual reproduction. In agreement with this, recent studies on invertebrates have shown a prevalence of asexual species in agricultural (simplified) but not in natural (complex) environments. We investigated whether the same correlation between reproductive mode and habitat can be found in different populations within one species. The parasitoid wasp Tetrastichus coeruleus forms an ideal model to test this question, since it occurs both in natural and agricultural environments. Further, we investigated whether Wolbachia infection caused parthenogenesis in female-biased populations. In contrast to the general pattern, in Dutch and French natural areas, we found Wolbachia-infected, highly female-biased populations that reproduce parthenogenetically. In contrast, populations on Dutch agricultural fields were not infected with Wolbachia, showed higher frequencies of males and reproduced sexually. However, we also found a female-only, Wolbachia-infected population on agricultural fields in north-eastern United States. All Wolbachia-infected populations were infected with the same Wolbachia strain. At this moment, we do not have a convincing explanation for this deviation from the general pattern of ecology and reproductive mode. It may be that asparagus agricultural fields differ from other crop fields in ways that favour sexual reproduction. Alternatively, Wolbachia may manipulate life history traits in its host, resulting in different fitness pay-offs in different habitats. The fixation of Wolbachia in the United States populations (where the species was introduced) may be due to founder effect and lack of uninfected, sexual source populations.}, } @article {pmid20300514, year = {2010}, author = {Tortosa, P and Charlat, S and Labbé, P and Dehecq, JS and Barré, H and Weill, M}, title = {Wolbachia age-sex-specific density in Aedes albopictus: a host evolutionary response to cytoplasmic incompatibility?.}, journal = {PloS one}, volume = {5}, number = {3}, pages = {e9700}, pmid = {20300514}, issn = {1932-6203}, mesh = {Aedes/*metabolism ; Animals ; Cytoplasm/*metabolism ; Evolution, Molecular ; Female ; France ; Genotype ; Geography ; Greece ; Male ; Models, Genetic ; Polymerase Chain Reaction/methods ; Sex Factors ; Time Factors ; Wolbachia/*metabolism ; }, abstract = {BACKGROUND: Wolbachia bacteria have invaded many arthropod species by inducing Cytoplasmic Incompatibility (CI). These symbionts represent fascinating objects of study for evolutionary biologists, but also powerful potential biocontrol agents. Here, we assess the density dynamics of Wolbachia infections in males and females of the mosquito Aedes albopitcus, an important vector of human pathogens, and interpret the results within an evolutionary framework.

Wolbachia densities were measured in natural populations and in age controlled mosquitoes using quantitative PCR. We show that the density dynamics of the wAlbA Wolbachia strain infecting Aedes albopictus drastically differ between males and females, with a very rapid decay of infection in males only.

CONCLUSIONS/SIGNIFICANCE: Theory predicts that Wolbachia and its hosts should cooperate to improve the transmission of infection to offspring, because only infected eggs are protected from the effects of CI. However, incompatible matings effectively lower the fertility of infected males, so that selection acting on the host genome should tend to reduce the expression of CI in males, for example, by reducing infection density in males before sexual maturation. The rapid decay of one Wolbachia infection in Aedes albopictus males, but not in females, is consistent with this prediction. We suggest that the commonly observed reduction in CI intensity with male age reflects a similar evolutionary process. Our results also highlight the importance of monitoring infection density dynamics in both males and females to assess the efficiency of Wolbachia-based control strategies.}, } @article {pmid20232169, year = {2010}, author = {Farkas, JZ and Hinow, P}, title = {Structured and unstructured continuous models for Wolbachia infections.}, journal = {Bulletin of mathematical biology}, volume = {72}, number = {8}, pages = {2067-2088}, doi = {10.1007/s11538-010-9528-1}, pmid = {20232169}, issn = {1522-9602}, mesh = {Animals ; Female ; Gram-Negative Bacterial Infections/*immunology ; Male ; *Models, Immunological ; Symbiosis ; Wolbachia/*immunology ; }, abstract = {We introduce and investigate a series of models for an infection of a diplodiploid host species by the bacterial endosymbiont Wolbachia. The continuous models are characterized by partial vertical transmission, cytoplasmic incompatibility and fitness costs associated with the infection. A particular aspect of interest is competitions between mutually incompatible strains. We further introduce an age-structured model that takes into account different fertility and mortality rates at different stages of the life cycle of the individuals. With only a few parameters, the ordinary differential equation models exhibit already interesting dynamics and can be used to predict criteria under which a strain of bacteria is able to invade a population. Interestingly, but not surprisingly, the age-structured model shows significant differences concerning the existence and stability of equilibrium solutions compared to the unstructured model.}, } @article {pmid20231892, year = {2010}, author = {Azpurua, J and De La Cruz, D and Valderama, A and Windsor, D}, title = {Lutzomyia sand fly diversity and rates of infection by Wolbachia and an exotic Leishmania species on Barro Colorado Island, Panama.}, journal = {PLoS neglected tropical diseases}, volume = {4}, number = {3}, pages = {e627}, pmid = {20231892}, issn = {1935-2735}, mesh = {Animals ; Cluster Analysis ; DNA, Protozoan/genetics ; Electron Transport Complex IV/genetics ; *Genetic Variation ; Insect Proteins/genetics ; Leishmania/genetics/*isolation & purification ; Molecular Sequence Data ; Panama ; Phylogeny ; Psychodidae/*classification/genetics/microbiology/parasitology ; RNA, Ribosomal, 18S/genetics ; Sequence Analysis, DNA ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Sand flies (Diptera, Psychodidae, Phlebotominae) in the genus Lutzomyia are the predominant vectors of the protozoan disease leishmaniasis in the New World. Within the watershed of the Panama Canal, the cutaneous form of leishmaniasis is a continuous health threat for residents, tourists and members of an international research community. Here we report the results of screening a tropical forest assemblage of sand fly species for infection by both Leishmania and a microbe that can potentially serve in vector population control, the cytoplasmically transmitted rickettsia, Wolbachia pipientis. Knowing accurately which Lutzomyia species are present, what their evolutionary relationships are, and how they are infected by strains of both Leishmania and Wolbachia is of critical value for building strategies to mitigate the impact of this disease in humans.

METHODOLOGY AND FINDINGS: We collected, sorted and then used DNA sequences to determine the diversity and probable phylogenetic relationships of the Phlebotominae occurring in the understory of Barro Colorado Island in the Republic of Panama. Sequence from CO1, the DNA barcoding gene, supported 18 morphology-based species determinations while revealing the presence of two possible "cryptic" species, one (Lu. sp. nr vespertilionis) within the Vespertilionis group, the other (Lu. gomezi) within the Lutzomyia-cruciata series. Using ITS-1 and "minicircle" primers we detected Leishmania DNA in 43.3% of Lu. trapidoi, 26.3% of Lu. gomezi individuals and in 0% of the other 18 sand fly species. Identical ITS-1 sequence was obtained from the Leishmania infecting Lu. trapidoi and Lu. gomezi, sequence which was 93% similar to Leishmania (viannia) naiffi in GenBank, a species previously unknown in Panama, but recognized as a type of cutaneous leishmaniasis vectored broadly across northern and central South America. Distinct strains of the intracellular bacterium Wolbachia were detected in three of 20 sand fly species, including Lu. trapidoi, in which it frequently co-occurred with Leishmania.

CONCLUSIONS: Both morphological and molecular methods were used to examine an assemblage of 20 sand fly species occurring in the forests of the Panama Canal area. Two of these species, members of separate clades, were found to carry Leishmania at high frequency and hence are likely vectors of leishmaniasis to humans or other mammal species. A single Leishmania species, identified with high confidence as Le. naiffi, was carried by both species. That Le. naiffi is known to cause cutaneous lesions in South America but has hitherto not been reported or implicated in Panama opens the possibility that its range has recently expanded to include the Isthmus or that it occurs as a recent introduction. The occurrence of Leishmania and Wolbachia in Lu. trapidoi identifies one important vector of the disease as a potential target for gene introductions using Wolbachia population sweeps.}, } @article {pmid20219740, year = {2010}, author = {Carrington, LB and Hoffmann, AA and Weeks, AR}, title = {Monitoring long-term evolutionary changes following Wolbachia introduction into a novel host: the Wolbachia popcorn infection in Drosophila simulans.}, journal = {Proceedings. Biological sciences}, volume = {277}, number = {1690}, pages = {2059-2068}, pmid = {20219740}, issn = {1471-2954}, mesh = {Animals ; *Biological Evolution ; Drosophila melanogaster/genetics/*microbiology ; Female ; *Host-Pathogen Interactions ; Longevity/*genetics ; Male ; Selection, Genetic ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia may act as a biological control agent for pest management; in particular, the Wolbachia variant wMelPop (popcorn) shortens host longevity and may be useful for dengue suppression. However, long-term changes in the host and Wolbachia genomes can alter Wolbachia spread and/or host effects that suppress disease. Here, we investigate the phenotypic effects of wMelPop in a non-native host, Drosophila simulans, following artificial transinfection approximately 200 generations ago. Long-term rearing and maintenance of the bacteria were at 19 degrees C in the original I-102 genetic background that was transinfected with the popcorn strain. The bacteria were then introgressed into three massbred backgrounds, and tetracycline was used to create uninfected sublines. The effect of wMelPop on longevity in this species appears to have changed; longevity was no longer reduced at 25 degrees C in some nuclear backgrounds, reflecting different geographical origin, selection or drift, although the reduction was still evident for flies held at 30 degrees C. Wolbachia influenced productivity and viability, and development time in some host backgrounds. These findings suggest that long-term attenuation of Wolbachia effects may compromise the effectiveness of this bacterium in pest control. They also emphasize the importance of host nuclear background on Wolbachia phenotypic effects.}, } @article {pmid20217091, year = {2010}, author = {Ahmed, MZ and Ren, SX and Xue, X and Li, XX and Jin, GH and Qiu, BL}, title = {Prevalence of endosymbionts in Bemisia tabaci populations and their in vivo sensitivity to antibiotics.}, journal = {Current microbiology}, volume = {61}, number = {4}, pages = {322-328}, pmid = {20217091}, issn = {1432-0991}, mesh = {Ampicillin/pharmacology ; Animals ; Anti-Bacterial Agents/pharmacology ; China ; DNA, Bacterial/analysis ; Enterobacteriaceae/*drug effects ; Halomonadaceae/*drug effects ; Hemiptera/*microbiology ; Plant Diseases ; Polymerase Chain Reaction ; Rifampin/pharmacology ; *Symbiosis ; Tetracycline/pharmacology ; Wolbachia/*drug effects ; }, abstract = {Bemisia tabaci can harbor both primary and secondary endosymbionts, and the specific endosymbionts can differ among different B. tabaci biotypes. This study determined (1) the prevalence of the primary endosymbiont Portiera aleyrodidarum and secondary endosymbionts Arsenophonus and Wolbachia in two invasive biotypes (B and Q) and one indigenous biotype (Cv) in China and (2) the in vivo effect of three antibiotics (tetracycline, ampicillin trihydrate, and rifampicin) against the endosymbionts; if an antibiotic substantially inhibits an endosymbiont, it could be used to determine the effect of that endosymbiont on B. tabaci. P. aleyrodidarum and Wolbachia were detected in all the three biotypes, while Arsenophonus was found only in the Q and Cv biotypes. P. aleyrodidarum was found in all tested individuals of the three biotypes. Infection rates of Wolbachia in the B, Cv, and Q biotypes were 58, 68, and 48%, respectively. The infection rate of Arsenophonus was 44% in the Q biotype but only 22% in the Cv biotype. The antibiotics failed to eliminate P. aleyrodidarum from any individual of the B, Cv, and Q biotypes but eliminated the secondary endosymbionts, Arsenophonus and Wolbachia, from 50 to 80% of the adult B. tabaci. The effect of the antibiotics depended on the species of endosymbiont, the antibiotic, the B. tabaci biotype, and various interactions between these factors. When used against Arsenophonus, the efficiency of rifampicin was better than ampicillin and tetracycline, regardless of B. tabaci biotype. When inactivating Wolbachia in Cv and Q biotypes, the efficiency tetracycline was better than ampicillin and rifampicin, and while the efficiency of tetracycline was better than rifampicin and ampicillin when they were used against Wolbachia in B biotype.}, } @article {pmid20199697, year = {2010}, author = {Brattig, NW and Hoerauf, A and Fischer, PU and Liebau, E and Bandi, C and Debrah, A and Büttner, M and Büttner, DW}, title = {Immunohistological studies on neoplasms of female and male Onchocerca volvulus: filarial origin and absence of Wolbachia from tumor cells.}, journal = {Parasitology}, volume = {137}, number = {5}, pages = {841-854}, pmid = {20199697}, issn = {1469-8161}, mesh = {Africa South of the Sahara ; Animals ; Doxycycline/therapeutic use ; Female ; Helminth Proteins/immunology/*isolation & purification ; Humans ; Immunohistochemistry ; Male ; Neoplasms/drug therapy/immunology/*parasitology ; Onchocerca volvulus/immunology/*isolation & purification ; Onchocerciasis/drug therapy/immunology/parasitology/*pathology ; }, abstract = {Up to 5% of untreated female Onchocerca volvulus filariae develop potentially fatal pleomorphic neoplasms, whose incidence is increased following ivermectin treatment. We studied the occurrence of 8 filarial proteins and of Wolbachia endobacteria in the tumor cells. Onchocercomas from patients, untreated and treated with antibiotics and anthelminthics, were examined by immunohistology. Neoplasms were diagnosed in 112 of 3587 female and in 2 of 1570 male O. volvulus. The following proteins and other compounds of O. volvulus were expressed in the cells of the neoplasms: glutathione S-transferase 1, lysosomal aspartic protease, cAMP-dependent protein kinase, alpha-enolase, aspartate aminotransferase, ankyrin E1, tropomyosin, heat shock protein 60, transforming growth factor-beta, and prostaglandin E(2). These findings prove the filarial origin of the neoplasms and confirm the pleomorphism of the tumor cells. Signs indicating malignancy of the neoplasms are described. Wolbachia were observed in the hypodermis, oocytes, and embryos of tumor-harbouring filariae using antibodies against Wolbachia surface protein, Wolbachia HtrA-type serine protease, and Wolbachia aspartate aminotransferase. In contrast, Wolbachia were not found in the cells of the neoplasms. Further, neoplasm-containing worms were not observed after more than 10 months after the start of sufficient treatment with doxycycline or doxycycline plus ivermectin.}, } @article {pmid20198874, year = {2010}, author = {Andrianova, BV and Goriacheva, II and Aleksandrov, ID and Gorelova, TV}, title = {[Establishment of a new continuous cell line of Drosophila melanogaster strain infected by the intracellular endosymbiotic bacterium Wolbachia pipientis under natural conditions].}, journal = {Genetika}, volume = {46}, number = {1}, pages = {14-17}, pmid = {20198874}, issn = {0016-6758}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Cell Line ; Drosophila melanogaster/*cytology/embryology/*microbiology ; Female ; Male ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {Wolbachia pipientis is an obligately intracellular bacterium infecting a number of arthropod and nematode species. At the body level, Wolbachia infection may cause parthenogenesis, feminization of genetic males, male killing, or cytoplasmic incompatibility; it may also be asymptomatic. Of special interest is DNA transfer from Wolbachia to the host insect genome, which was discovered recently. At the cellular level, the effects caused by Wolbachia have been studied more poorly. Only one of the known insect cell lines has been obtained from an insect species (the mosquito Aedes albopictus) infected by Wolbachia. In this study, a continuous cell line Dm2008Wb1 has been obtained from embryos of Drosophila melanogaster infected under natural conditions. Wolbachia both persists in a primary cell culture and is retained upon its transformation into a continuous culture. The presence of this bacterium in cells in a free form is evidenced by the fact that tetracycline treatment can cure the cells of Wolbachia and by successful transfer of Wolbachia to another cell line (S2), where it has not been detected before.}, } @article {pmid20195512, year = {2010}, author = {Grönke, S and Clarke, DF and Broughton, S and Andrews, TD and Partridge, L}, title = {Molecular evolution and functional characterization of Drosophila insulin-like peptides.}, journal = {PLoS genetics}, volume = {6}, number = {2}, pages = {e1000857}, pmid = {20195512}, issn = {1553-7404}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Body Weight/drug effects ; Diet ; Drosophila/drug effects/*genetics/growth & development/microbiology ; Drosophila Proteins/*genetics/*metabolism ; Drug Resistance/drug effects ; Energy Metabolism/drug effects/genetics ; *Evolution, Molecular ; Fertility/drug effects ; Gene Expression Regulation, Developmental/drug effects ; Genetic Loci/genetics ; Longevity/drug effects ; Mutation/genetics ; Ovum/cytology/drug effects ; Phylogeny ; Stress, Physiological/drug effects/genetics ; Survival Analysis ; Time Factors ; Wolbachia/metabolism ; Xenobiotics/pharmacology ; }, abstract = {Multicellular animals match costly activities, such as growth and reproduction, to the environment through nutrient-sensing pathways. The insulin/IGF signaling (IIS) pathway plays key roles in growth, metabolism, stress resistance, reproduction, and longevity in diverse organisms including mammals. Invertebrate genomes often contain multiple genes encoding insulin-like ligands, including seven Drosophila insulin-like peptides (DILPs). We investigated the evolution, diversification, redundancy, and functions of the DILPs, combining evolutionary analysis, based on the completed genome sequences of 12 Drosophila species, and functional analysis, based on newly-generated knock-out mutations for all 7 dilp genes in D. melanogaster. Diversification of the 7 DILPs preceded diversification of Drosophila species, with stable gene diversification and family membership, suggesting stabilising selection for gene function. Gene knock-outs demonstrated both synergy and compensation of expression between different DILPs, notably with DILP3 required for normal expression of DILPs 2 and 5 in brain neurosecretory cells and expression of DILP6 in the fat body compensating for loss of brain DILPs. Loss of DILP2 increased lifespan and loss of DILP6 reduced growth, while loss of DILP7 did not affect fertility, contrary to its proposed role as a Drosophila relaxin. Importantly, loss of DILPs produced in the brain greatly extended lifespan but only in the presence of the endosymbiontic bacterium Wolbachia, demonstrating a specific interaction between IIS and Wolbachia in lifespan regulation. Furthermore, loss of brain DILPs blocked the responses of lifespan and fecundity to dietary restriction (DR) and the DR response of these mutants suggests that IIS extends lifespan through mechanisms that both overlap with those of DR and through additional mechanisms that are independent of those at work in DR. Evolutionary conservation has thus been accompanied by synergy, redundancy, and functional differentiation between DILPs, and these features may themselves be of evolutionary advantage.}, } @article {pmid20195500, year = {2010}, author = {Nikoh, N and McCutcheon, JP and Kudo, T and Miyagishima, SY and Moran, NA and Nakabachi, A}, title = {Bacterial genes in the aphid genome: absence of functional gene transfer from Buchnera to its host.}, journal = {PLoS genetics}, volume = {6}, number = {2}, pages = {e1000827}, pmid = {20195500}, issn = {1553-7404}, support = {K12 GM000708/GM/NIGMS NIH HHS/United States ; 1K 12 GM00708/GM/NIGMS NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Aphids/*genetics/*microbiology ; Bacterial Proteins/genetics ; Buchnera/enzymology/*genetics ; Carboxypeptidases/genetics ; DNA Polymerase III/genetics ; Eukaryotic Cells/metabolism ; Gene Duplication ; Gene Fusion ; Gene Transfer, Horizontal/*genetics ; Genes, Bacterial/*genetics ; Genome ; Glycoside Hydrolases/chemistry/genetics ; Host-Pathogen Interactions/*genetics ; Molecular Sequence Data ; Muramidase/genetics ; N-Acetylmuramoyl-L-alanine Amidase/chemistry/genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; Rickettsia/genetics ; }, abstract = {Genome reduction is typical of obligate symbionts. In cellular organelles, this reduction partly reflects transfer of ancestral bacterial genes to the host genome, but little is known about gene transfer in other obligate symbioses. Aphids harbor anciently acquired obligate mutualists, Buchnera aphidicola (Gammaproteobacteria), which have highly reduced genomes (420-650 kb), raising the possibility of gene transfer from ancestral Buchnera to the aphid genome. In addition, aphids often harbor other bacteria that also are potential sources of transferred genes. Previous limited sampling of genes expressed in bacteriocytes, the specialized cells that harbor Buchnera, revealed that aphids acquired at least two genes from bacteria. The newly sequenced genome of the pea aphid, Acyrthosiphon pisum, presents the first opportunity for a complete inventory of genes transferred from bacteria to the host genome in the context of an ancient obligate symbiosis. Computational screening of the entire A. pisum genome, followed by phylogenetic and experimental analyses, provided strong support for the transfer of 12 genes or gene fragments from bacteria to the aphid genome: three LD-carboxypeptidases (LdcA1, LdcA2,psiLdcA), five rare lipoprotein As (RlpA1-5), N-acetylmuramoyl-L-alanine amidase (AmiD), 1,4-beta-N-acetylmuramidase (bLys), DNA polymerase III alpha chain (psiDnaE), and ATP synthase delta chain (psiAtpH). Buchnera was the apparent source of two highly truncated pseudogenes (psiDnaE and psiAtpH). Most other transferred genes were closely related to genes from relatives of Wolbachia (Alphaproteobacteria). At least eight of the transferred genes (LdcA1, AmiD, RlpA1-5, bLys) appear to be functional, and expression of seven (LdcA1, AmiD, RlpA1-5) are highly upregulated in bacteriocytes. The LdcAs and RlpAs appear to have been duplicated after transfer. Our results excluded the hypothesis that genome reduction in Buchnera has been accompanied by gene transfer to the host nuclear genome, but suggest that aphids utilize a set of duplicated genes acquired from other bacteria in the context of the Buchnera-aphid mutualism.}, } @article {pmid20193981, year = {2010}, author = {Dingman, P and Levy, JK and Kramer, LH and Johnson, CM and Lappin, MR and Greiner, EC and Courtney, CH and Tucker, SJ and Morchon, R}, title = {Association of Wolbachia with heartworm disease in cats and dogs.}, journal = {Veterinary parasitology}, volume = {170}, number = {1-2}, pages = {50-60}, doi = {10.1016/j.vetpar.2010.01.037}, pmid = {20193981}, issn = {1873-2550}, mesh = {Animals ; Anti-Bacterial Agents ; Antibodies, Bacterial/blood ; Antibodies, Helminth/blood ; Cat Diseases/drug therapy/immunology/*microbiology/*parasitology ; Cats ; DNA, Bacterial/chemistry/genetics ; Dirofilaria immitis/immunology/*microbiology ; Dirofilariasis/immunology/*parasitology ; Dog Diseases/drug therapy/immunology/*microbiology/*parasitology ; Dogs ; Immunohistochemistry/veterinary ; Lung Diseases, Parasitic/drug therapy/immunology/microbiology/parasitology/*veterinary ; Polymerase Chain Reaction/veterinary ; Statistics, Nonparametric ; Wolbachia/*immunology ; }, abstract = {Although the presence of adult Dirofilaria immitis in the pulmonary arteries and its associated arteritis and thromboembolic disease can explain some of the manifestations of canine and feline heartworm disease, the cause of other findings remains unclear. Cats with D. immitis antibodies but lacking adult parasites in the pulmonary arteries frequently develop histological lesions of the airways, resulting in a condition termed Heartworm-Associated Respiratory Disease. All D. immitis parasites harbor Wolbachia pipientis bacteria and D. immitis-infected animals can have circulating Wolbachia antibodies and pro-inflammatory Wolbachia antigens (WSP) deposited in tissues. Little is known about the role that Wolbachia plays in lung disease of animals naturally infected with D. immitis. The purpose of this study was to determine the contribution of Wolbachia to the pathogenesis of natural heartworm disease in cats and dogs. We hypothesized that animals having sufficient Wolbachia burden to be detected in lung tissue by immunohistochemistry and/or polymerase chain reaction (PCR) would have more severe pulmonary disease than those with bacteria below the limits of detection. We further hypothesized that animals that were immunoreactive to pro-inflammatory WSP would have more severe pulmonary lesions than those that were seronegative for WSP antibodies. Blood and lung tissue samples were collected from cats and dogs representing three different D. immitis infection statuses: heartworm-free, heartworm-exposed, heartworm-infected. There was a positive but weak correlation between the magnitude of D. immitis antibody titers and WSP titers in cats (r=0.57, p<0.001) and in dogs (r=0.39, p<0.001). Pulmonary lesions were more common in HW-infected animals than in HW-free animals. Pulmonary arteriolar occlusion was more common in HW-infected cats (57%; p=0.003) than in HW-infected dogs (17%). Although pulmonary lesions were most common in HW-infected animals, there was no clear additive effect when either Wolbachia DNA/WSP was detected in lung tissue or when circulating Wolbachia antibodies were detected. There were no significant differences in the magnitude of pulmonary lesion scores within each HW-infection status group regardless of whether Wolbachia DNA/WSP or antibodies were detected. The relationship between Wolbachia and lung pathology in heartworm-infected animals remains to be determined. The lack of clear evidence for a role of Wolbachia in heartworm disease creates a dilemma for veterinarians treating animals in D. immitis-endemic areas. Although the indiscriminant use of antibiotics should be avoided, many clinicians prescribe doxycycline based on the favorable responses observed in human filarial diseases and promising results from the first published studies of doxycycline use in D. immitis-infected dogs.}, } @article {pmid20167019, year = {2010}, author = {Darby, AC and Choi, JH and Wilkes, T and Hughes, MA and Werren, JH and Hurst, GD and Colbourne, JK}, title = {Characteristics of the genome of Arsenophonus nasoniae, son-killer bacterium of the wasp Nasonia.}, journal = {Insect molecular biology}, volume = {19 Suppl 1}, number = {}, pages = {75-89}, doi = {10.1111/j.1365-2583.2009.00950.x}, pmid = {20167019}, issn = {1365-2583}, support = {//Medical Research Council/United Kingdom ; }, mesh = {Animals ; Base Composition/genetics ; Base Sequence ; Bayes Theorem ; Enterobacteriaceae/*genetics ; Genes, Bacterial/genetics ; Genome, Bacterial/*genetics ; Genomics ; Models, Genetic ; Molecular Sequence Data ; Open Reading Frames/genetics ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Signal Transduction/genetics ; Symbiosis/*genetics ; Wasps/*microbiology ; }, abstract = {We report the properties of a draft genome sequence of the bacterium Arsenophonus nasoniae, son-killer bacterium of Nasonia vitripennis. The genome sequence data from this study are the first for a male-killing bacterium, and represent a microorganism that is unusual compared with other sequenced symbionts, in having routine vertical and horizontal transmission, two alternating hosts, and being culturable on cell-free media. The resulting sequence totals c. 3.5 Mbp and is annotated to contain 3332 predicted open reading frames (ORFs). Therefore, Arsenophonus represents a relatively large genome for an insect symbiont. The annotated ORF set suggests that the microbe is capable of a broad array of metabolic functions, well beyond those found for reproductive parasite genomes sequenced to date and more akin to horizontally transmitted and secondary symbionts. We also find evidence of genetic transfer from Wolbachia symbionts, and phage exchange with other gammaproteobacterial symbionts. These findings reflect the complex biology of a bacterium that is able to live, invade and survive multiple host environments while resisting immune responses.}, } @article {pmid20163713, year = {2010}, author = {Baldo, L and Desjardins, CA and Russell, JA and Stahlhut, JK and Werren, JH}, title = {Accelerated microevolution in an outer membrane protein (OMP) of the intracellular bacteria Wolbachia.}, journal = {BMC evolutionary biology}, volume = {10}, number = {}, pages = {48}, pmid = {20163713}, issn = {1471-2148}, mesh = {Animals ; Arthropods/immunology/microbiology/physiology ; Bacterial Outer Membrane Proteins/chemistry/*genetics/immunology ; *Evolution, Molecular ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Outer membrane proteins (OMPs) of Gram-negative bacteria are key players in the biology of bacterial-host interactions. However, while considerable attention has been given to OMPs of vertebrate pathogens, relatively little is known about the role of these proteins in bacteria that primarily infect invertebrates. One such OMP is found in the intracellular bacteria Wolbachia, which are widespread symbionts of arthropods and filarial nematodes. Recent experimental studies have shown that the Wolbachia surface protein (WSP) can trigger host immune responses and control cell death programming in humans, suggesting a key role of WSP for establishment and persistence of the symbiosis in arthropods.

RESULTS: Here we performed an analysis of 515 unique alleles found in 831 Wolbachia isolates, to investigate WSP structure, microevolution and population genetics. WSP shows an eight-strand transmembrane beta-barrel structure with four extracellular loops containing hypervariable regions (HVRs). A clustering approach based upon patterns of HVR haplotype diversity was used to group similar WSP sequences and to estimate the relative contribution of mutation and recombination during early stages of protein divergence. Results indicate that although point mutations generate most of the new protein haplotypes, recombination is a predominant force triggering diversity since the very first steps of protein evolution, causing at least 50% of the total amino acid variation observed in recently diverged proteins. Analysis of synonymous variants indicates that individual WSP protein types are subject to a very rapid turnover and that HVRs can accommodate a virtually unlimited repertoire of peptides. Overall distribution of WSP across hosts supports a non-random association of WSP with the host genus, although extensive horizontal transfer has occurred also in recent times.

CONCLUSIONS: In OMPs of vertebrate pathogens, large recombination impact, positive selection, reduced structural and compositional constraints, and extensive lateral gene transfer are considered hallmarks of evolution in response to the adaptive immune system. However, Wolbachia do not infect vertebrates. Here we predict that the rapid turnover of WSP loop motifs could aid in evading or inhibiting the invertebrate innate immune response. Overall, these features identify WSP as a strong candidate for future studies of host-Wolbachia interactions that affect establishment and persistence of this widespread endosymbiosis.}, } @article {pmid20147057, year = {2009}, author = {Werren, JH and Loehlin, DW}, title = {Curing Wolbachia infections in Nasonia (parasitoid wasp).}, journal = {Cold Spring Harbor protocols}, volume = {2009}, number = {10}, pages = {pdb.prot5312}, pmid = {20147057}, issn = {1559-6095}, support = {R24 GM084917/GM/NIGMS NIH HHS/United States ; R24 GM084917-02/GM/NIGMS NIH HHS/United States ; 1 R24 GM084917-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/chemistry ; Biology/methods ; Cytoplasm/metabolism ; Genome ; Male ; Species Specificity ; Spermatozoa/metabolism ; Wasps/genetics/*microbiology ; Wolbachia/*metabolism ; }, abstract = {Nasonia is a complex of four closely related species of wasps with several features that make it an excellent system for a variety of genetic studies. These include a short generation time, ease of rearing, interfertile species, visible and molecular markers, and a sequenced genome. However, the Nasonia species normally are completely or partially reproductively isolated owing to Wolbachia, widespread intracellular bacteria found in arthropods and nematodes. Wolbachia cause cytoplasmic incompatibility (CI) in Nasonia. Although the current genome-sequenced strains of Nasonia (AsymCX, IV7[u], RV2X[u]) are all Wolbachia-free, crosses between infected and uninfected wasps, or between infected wasps of different species, will fail because of Wolbachia. The sperm modification that induces CI appears to be established before the adult stage, so it is generally not possible to cure an infection and break the CI barrier in the same generation. However, a Wolbachia infection can be removed from a strain over a number of generations by feeding antibiotics and selecting on females showing partial CI. This protocol describes the methods to cure a Nasonia strain of its Wolbachia symbionts.}, } @article {pmid20147035, year = {2009}, author = {Werren, JH and Loehlin, DW}, title = {The parasitoid wasp Nasonia: an emerging model system with haploid male genetics.}, journal = {Cold Spring Harbor protocols}, volume = {2009}, number = {10}, pages = {pdb.emo134}, pmid = {20147035}, issn = {1559-6095}, support = {R24 GM084917/GM/NIGMS NIH HHS/United States ; R24 GM084917-02/GM/NIGMS NIH HHS/United States ; 1 R24 GM084917-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Diploidy ; Female ; Genome ; Genomics ; *Haploidy ; Insecta ; Male ; Microsatellite Repeats ; Models, Genetic ; Polymorphism, Single Nucleotide ; Quantitative Trait Loci ; Sexual Behavior, Animal ; Wasps/*genetics/*physiology ; }, abstract = {Nasonia is a complex of four closely related species of wasps that is rapidly emerging as a model for evolutionary and developmental genetics. It has several features that make it an excellent genetic system, including a short generation time, ease of rearing, interfertile species, visible and molecular markers, and a sequenced genome. The form of sex determination, called "haplodiploidy," makes Nasonia particularly suitable as a genetic tool. Females are diploid and develop from fertilized eggs, whereas males are haploid and develop from unfertilized eggs. This allows geneticists to exploit many of the advantages of haploid genetics in an otherwise complex eukaryotic organism. Nasonia readily inbreeds, permitting production of isogenic lines, and the four species in the genus are interfertile (after removal of the endosymbiont Wolbachia), facilitating movement of genes between the species for efficient positional cloning of quantitative trait loci (QTL). Genome sequencing of the genetic model Nasonia vitripennis and two interfertile species, Nasonia giraulti and Nasonia longicornis, is now completed. This genome project provides a wealth of interspecies polymorphisms (e.g., single nucleotide polymorphisms [SNPs], insertion-deletions [indels], microsatellites) to facilitate positional cloning of genes involved in species differences in behavior, morphology, and development. Advances in the genetics of this system also open a path for improvement of parasitoid insects as agents of pest control.}, } @article {pmid20097822, year = {2010}, author = {Tang, M and Lv, L and Jing, S and Zhu, L and He, G}, title = {Bacterial symbionts of the brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae).}, journal = {Applied and environmental microbiology}, volume = {76}, number = {6}, pages = {1740-1745}, pmid = {20097822}, issn = {1098-5336}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; *Bacterial Physiological Phenomena ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Gastrointestinal Tract/microbiology ; Hemiptera/*microbiology ; In Situ Hybridization ; Molecular Sequence Data ; Oryza/parasitology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {The brown planthopper (Nilaparvata lugens Stål), the most destructive pest of rice, has been identified, including biotypes with high virulence towards previously resistant rice varieties. There have also been many reports of a yeast-like symbiont of N. lugens, but little is known about the bacterial microbes. In this study, we examined the bacterial microbes in N. lugens and identified a total of 18 operational taxonomic units (OTUs) representing four phyla (Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes) by sequencing and analyzing 16S rRNA gene libraries obtained from three populations of N. lugens, which were maintained on the rice varieties TN1, Mudgo, and ASD7. Several of the OTUs were similar to previously reported secondary symbionts of other insects, including an endosymbiont of the psyllid Glycapsis brimblecombei, an Asaia sp. found in the mosquito Anopheles stephensi, and Wolbachia, found in the mite Metaseiulus occidentalis. However, the species and numbers of the detected OTUs differed substantially among the N. lugens populations. Further, in situ hybridization analysis using digoxigenin-labeled probes indicated that OTU 1 was located in hypogastrium tissues near the ovipositor and ovary in biotype 1 insects, while OTU 2 was located in the front of the ovipositor sheath in biotype 2 insects. In addition, masses of bacterium-like organisms were observed in the tubes of salivary sheaths in rice plant tissues that the insects had fed upon. The results provide indications of the diversity of the bacterial microbes harbored by the brown planthopper and of possible associations between specific bacterial microbes and biotypes of N. lugens.}, } @article {pmid20089982, year = {2010}, author = {Raoult, D}, title = {Doxycycline for Mansonella perstans infection.}, journal = {The New England journal of medicine}, volume = {362}, number = {3}, pages = {272; author reply 272-3}, doi = {10.1056/NEJMc0910675}, pmid = {20089982}, issn = {1533-4406}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Drug Resistance ; Drug Therapy, Combination ; Elephantiasis, Filarial/drug therapy ; Filaricides/*therapeutic use ; Humans ; Mansonella/*drug effects/genetics ; Mansonelliasis/complications/*drug therapy ; Rickettsiaceae Infections/complications/*drug therapy ; Wolbachia/genetics ; Wuchereria bancrofti/isolation & purification ; }, } @article {pmid20087396, year = {2010}, author = {Raychoudhury, R and Grillenberger, BK and Gadau, J and Bijlsma, R and van de Zande, L and Werren, JH and Beukeboom, LW}, title = {Phylogeography of Nasonia vitripennis (Hymenoptera) indicates a mitochondrial-Wolbachia sweep in North America.}, journal = {Heredity}, volume = {104}, number = {3}, pages = {318-326}, doi = {10.1038/hdy.2009.160}, pmid = {20087396}, issn = {1365-2540}, mesh = {Animals ; Bacterial Proteins/genetics ; Evolution, Molecular ; Genetic Variation ; Insect Proteins/genetics ; Microsatellite Repeats ; Mitochondria/*genetics/microbiology ; Models, Genetic ; Molecular Sequence Data ; Mutation ; North America ; *Phylogeny ; Wasps/*classification/genetics/microbiology ; Wolbachia/*genetics/physiology ; }, abstract = {Here we report evidence of a mitochondrial-Wolbachia sweep in North American populations of the parasitoid wasp Nasonia vitripennis, a cosmopolitan species and emerging model organism for evolutionary and genetic studies. Analysis of the genetic variation of 89 N. vitripennis specimens from Europe and North America was performed using four types of genetic markers: a portion of the mitochondrial cytochrome oxidase I gene, nine polymorphic nuclear microsatellites, sequences from 11 single-copy nuclear markers and six Wolbachia genes. The results show that the European populations have a sevenfold higher mitochondrial sequence variation than North American populations, but similar levels of microsatellite and nuclear gene sequence variation. Variation in the North American mitochondria is extremely low (pi=0.31%), despite a highly elevated mutation rate (approximately 35-40 times higher than the nuclear genes) in the mitochondria of Nasonia. The data are indicative of a mitochondrial sweep in the North American population, possibly due to Wolbachia infections that are maternally co-inherited with the mitochondria. Owing to similar levels of nuclear variation, the data could not resolve whether N. vitripennis originated in the New or the Old World.}, } @article {pmid20087395, year = {2010}, author = {Clark, ME and O'Hara, FP and Chawla, A and Werren, JH}, title = {Behavioral and spermatogenic hybrid male breakdown in Nasonia.}, journal = {Heredity}, volume = {104}, number = {3}, pages = {289-301}, pmid = {20087395}, issn = {1365-2540}, support = {R01 GM070026/GM/NIGMS NIH HHS/United States ; R01 GM070026-04/GM/NIGMS NIH HHS/United States ; R01 GM0700026-04/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Chimera/genetics/*physiology ; Courtship ; Female ; Infertility ; Male ; Sexual Behavior, Animal ; *Spermatogenesis ; Spermatozoa/*cytology ; Wasps/genetics/*physiology ; }, abstract = {Several reproductive barriers exist within the Nasonia species complex, including allopatry, premating behavioral isolation, postzygotic inviability and Wolbachia-induced cytoplasmic incompatibility. Here we show that hybrid males suffer two additional reproductive disadvantages, an inability to properly court females and decreased sperm production. Hybrid behavioral sterility, characterized by a reduced ability of hybrids to perform necessary courtship behaviors, occurs in hybrids between two species of Nasonia. Hybrid males produced in crosses between N. vitripennis and N. giraulti courted females at a reduced frequency (23-69%), compared with wild-type N. vitripennis and N. giraulti males (>93%). Reduced courtship frequency was not a simple function of inactivity among hybrids. A strong effect of cytoplasmic (mitochondrial) background was also found in N. vitripennis and N. giraulti crosses; F2 hybrids with giraulti cytoplasm showing reduced ability at most stages of courtship. Hybrids produced between a younger species pair, N. giraulti and N. longicornis, were behaviorally fertile. All males possessed motile sperm, but sperm production is greatly reduced in hybrids between the older species pair, N. vitripennis and N. giraulti. This effect on hybrid males, lowered sperm counts rather than nonfunctional sperm, is different from most described cases of hybrid male sterility, and may represent an earlier stage of hybrid sperm breakdown. The results add to previous studies of F2 hybrid inviability and behavioral sterility, and indicate that Wolbachia-induced hybrid incompatibility has arisen early in species divergence, relative to behavioral sterility and spermatogenic infertility.}, } @article {pmid20083406, year = {2010}, author = {Kent, BN and Bordenstein, SR}, title = {Phage WO of Wolbachia: lambda of the endosymbiont world.}, journal = {Trends in microbiology}, volume = {18}, number = {4}, pages = {173-181}, pmid = {20083406}, issn = {1878-4380}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; R01 GM085163-02/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/microbiology/physiology ; Bacteriophage lambda/genetics/physiology ; Bacteriophages/genetics/*physiology ; Evolution, Molecular ; *Symbiosis ; Wolbachia/genetics/*physiology/*virology ; }, abstract = {The discovery of an extraordinarily high level of mobile elements in the genome of Wolbachia, a widespread arthropod and nematode endosymbiont, suggests that this bacterium could be an excellent model for assessing the evolution and function of mobile DNA in specialized bacteria. In this paper, we discuss how studies on the temperate bacteriophage WO of Wolbachia have revealed unexpected levels of genomic flux and are challenging previously held views about the clonality of obligate intracellular bacteria. We also discuss the roles this phage might play in the Wolbachia-arthropod symbiosis and infer how this research can be translated to combating human diseases vectored by arthropods. We expect that this temperate phage will be a preeminent model system to understand phage genetics, evolution and ecology in obligate intracellular bacteria. In this sense, phage WO might be likened to phage lambda of the endosymbiont world.}, } @article {pmid20081357, year = {2010}, author = {Haselkorn, TS}, title = {The Spiroplasma heritable bacterial endosymbiont of Drosophila.}, journal = {Fly}, volume = {4}, number = {1}, pages = {80-87}, doi = {10.4161/fly.4.1.10883}, pmid = {20081357}, issn = {1933-6942}, mesh = {Animals ; Bacterial Infections/transmission ; Biodiversity ; Biological Evolution ; Drosophila/*microbiology ; Spiroplasma/*physiology ; *Symbiosis ; }, abstract = {Since the discovery of the small, gram-positive bacteria, Spiroplasma, as a sex-ratio distorting agent in Drosophila over 50 years ago, substantial progress has been made in understanding the relationship of this bacteria with its insect host. Thus far, Spiroplasma have been found as heritable endosymbionts in sixteen different species of Drosophila. In some species these bacteria cause a male-killing phenotype, where the males die during embryogenesis. In other species, however, Spiroplasma does not cause male-killing, and its fitness effects are unclear. Though recent research has identified multiple factors that affect the prevalence and transmission of Spiroplasma in Drosophila populations, much work remains to fully characterize this symbiosis. Spiroplasma is the only identified heritable bacterial endosymbiont of Drosophila, other than Wolbachia, and can serve as a useful as model for elucidating the nature of insect/bacterial interactions.}, } @article {pmid20080750, year = {2010}, author = {Hosokawa, T and Koga, R and Kikuchi, Y and Meng, XY and Fukatsu, T}, title = {Wolbachia as a bacteriocyte-associated nutritional mutualist.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {107}, number = {2}, pages = {769-774}, pmid = {20080750}, issn = {1091-6490}, mesh = {Animal Feed ; Animals ; Anti-Bacterial Agents/pharmacology ; Aphids/microbiology/physiology ; Base Sequence ; Bedbugs/microbiology ; Biological Evolution ; DNA Primers ; Dietary Supplements ; Electrophoresis, Gel, Pulsed-Field ; Female ; Fertility ; *Genome, Bacterial ; Male ; Molecular Sequence Data ; Nucleic Acid Hybridization ; Phylogeny ; Polymerase Chain Reaction ; Symbiosis ; Tsetse Flies/microbiology/physiology ; Wolbachia/classification/drug effects/genetics/*physiology ; }, abstract = {Many insects are dependent on bacterial symbionts that provide essential nutrients (ex. aphid-Buchnera and tsetse-Wiglesworthia associations), wherein the symbionts are harbored in specific cells called bacteriocytes that constitute a symbiotic organ bacteriome. Facultative and parasitic bacterial symbionts like Wolbachia have been regarded as evolutionarily distinct from such obligate nutritional mutualists. However, we discovered that, in the bedbug Cimex lectularius, Wolbachia resides in a bacteriome and appears to be an obligate nutritional mutualist. Two bacterial symbionts, a Wolbachia strain and an unnamed gamma-proteobacterium, were identified from different strains of the bedbug. The Wolbachia symbiont was detected from all of the insects examined whereas the gamma-proteobacterium was found in a part of them. The Wolbachia symbiont was specifically localized in the bacteriomes and vertically transmitted via the somatic stem cell niche of germalia to oocytes, infecting the incipient symbiotic organ at an early stage of the embryogenesis. Elimination of the Wolbachia symbiont resulted in retarded growth and sterility of the host insect. These deficiencies were rescued by oral supplementation of B vitamins, confirming the essential nutritional role of the symbiont for the host. The estimated genome size of the Wolbachia symbiont was around 1.3 Mb, which was almost equivalent to the genome sizes of parasitic Wolbachia strains of other insects. These results indicate that bacteriocyte-associated nutritional mutualism can evolve from facultative and prevalent microbial associates like Wolbachia, highlighting a previously unknown aspect of the parasitism-mutualism evolutionary continuum.}, } @article {pmid20075255, year = {2010}, author = {Werren, JH and Richards, S and Desjardins, CA and Niehuis, O and Gadau, J and Colbourne, JK and , and Werren, JH and Richards, S and Desjardins, CA and Niehuis, O and Gadau, J and Colbourne, JK and Beukeboom, LW and Desplan, C and Elsik, CG and Grimmelikhuijzen, CJ and Kitts, P and Lynch, JA and Murphy, T and Oliveira, DC and Smith, CD and van de Zande, L and Worley, KC and Zdobnov, EM and Aerts, M and Albert, S and Anaya, VH and Anzola, JM and Barchuk, AR and Behura, SK and Bera, AN and Berenbaum, MR and Bertossa, RC and Bitondi, MM and Bordenstein, SR and Bork, P and Bornberg-Bauer, E and Brunain, M and Cazzamali, G and Chaboub, L and Chacko, J and Chavez, D and Childers, CP and Choi, JH and Clark, ME and Claudianos, C and Clinton, RA and Cree, AG and Cristino, AS and Dang, PM and Darby, AC and de Graaf, DC and Devreese, B and Dinh, HH and Edwards, R and Elango, N and Elhaik, E and Ermolaeva, O and Evans, JD and Foret, S and Fowler, GR and Gerlach, D and Gibson, JD and Gilbert, DG and Graur, D and Gründer, S and Hagen, DE and Han, Y and Hauser, F and Hultmark, D and Hunter, HC and Hurst, GD and Jhangian, SN and Jiang, H and Johnson, RM and Jones, AK and Junier, T and Kadowaki, T and Kamping, A and Kapustin, Y and Kechavarzi, B and Kim, J and Kim, J and Kiryutin, B and Koevoets, T and Kovar, CL and Kriventseva, EV and Kucharski, R and Lee, H and Lee, SL and Lees, K and Lewis, LR and Loehlin, DW and Logsdon, JM and Lopez, JA and Lozado, RJ and Maglott, D and Maleszka, R and Mayampurath, A and Mazur, DJ and McClure, MA and Moore, AD and Morgan, MB and Muller, J and Munoz-Torres, MC and Muzny, DM and Nazareth, LV and Neupert, S and Nguyen, NB and Nunes, FM and Oakeshott, JG and Okwuonu, GO and Pannebakker, BA and Pejaver, VR and Peng, Z and Pratt, SC and Predel, R and Pu, LL and Ranson, H and Raychoudhury, R and Rechtsteiner, A and Reese, JT and Reid, JG and Riddle, M and Robertson, HM and Romero-Severson, J and Rosenberg, M and Sackton, TB and Sattelle, DB and Schlüns, H and Schmitt, T and Schneider, M and Schüler, A and Schurko, AM and Shuker, DM and Simões, ZL and Sinha, S and Smith, Z and Solovyev, V and Souvorov, A and Springauf, A and Stafflinger, E and Stage, DE and Stanke, M and Tanaka, Y and Telschow, A and Trent, C and Vattathil, S and Verhulst, EC and Viljakainen, L and Wanner, KW and Waterhouse, RM and Whitfield, JB and Wilkes, TE and Williamson, M and Willis, JH and Wolschin, F and Wyder, S and Yamada, T and Yi, SV and Zecher, CN and Zhang, L and Gibbs, RA}, title = {Functional and evolutionary insights from the genomes of three parasitoid Nasonia species.}, journal = {Science (New York, N.Y.)}, volume = {327}, number = {5963}, pages = {343-348}, pmid = {20075255}, issn = {1095-9203}, support = {5R24GM084917-02/GM/NIGMS NIH HHS/United States ; R01 GM070026-04S1/GM/NIGMS NIH HHS/United States ; R24 GM084917/GM/NIGMS NIH HHS/United States ; R01 GM064864-05A2/GM/NIGMS NIH HHS/United States ; R01 GM070026/GM/NIGMS NIH HHS/United States ; R01 HG000747-14/HG/NHGRI NIH HHS/United States ; U54 HG003273-03/HG/NHGRI NIH HHS/United States ; R01 HG000747/HG/NHGRI NIH HHS/United States ; R01 GM064864/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; R01GM064864/GM/NIGMS NIH HHS/United States ; R01 GM085233/GM/NIGMS NIH HHS/United States ; R01 AI055624/AI/NIAID NIH HHS/United States ; U54 HG003273/HG/NHGRI NIH HHS/United States ; R24 GM084917-02/GM/NIGMS NIH HHS/United States ; 5R01HG000747-14/HG/NHGRI NIH HHS/United States ; /ImNIH/Intramural NIH HHS/United States ; R01 GM064864-04/GM/NIGMS NIH HHS/United States ; R01 GM085163/GM/NIGMS NIH HHS/United States ; R01 GM079484/GM/NIGMS NIH HHS/United States ; R24 GM084917-01/GM/NIGMS NIH HHS/United States ; 5R01GM070026-04/GM/NIGMS NIH HHS/United States ; AI028309-13A2/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Arthropods/parasitology ; *Biological Evolution ; DNA Methylation ; DNA Transposable Elements ; Female ; Gene Transfer, Horizontal ; Genes, Insect ; Genetic Speciation ; Genetic Variation ; *Genome, Insect ; Host-Parasite Interactions ; Insect Proteins/genetics/metabolism ; Insect Viruses/genetics ; Insecta/genetics ; Male ; Molecular Sequence Data ; Quantitative Trait Loci ; Recombination, Genetic ; Sequence Analysis, DNA ; Wasp Venoms/chemistry/toxicity ; Wasps/*genetics/physiology ; Wolbachia/genetics ; }, abstract = {We report here genome sequences and comparative analyses of three closely related parasitoid wasps: Nasonia vitripennis, N. giraulti, and N. longicornis. Parasitoids are important regulators of arthropod populations, including major agricultural pests and disease vectors, and Nasonia is an emerging genetic model, particularly for evolutionary and developmental genetics. Key findings include the identification of a functional DNA methylation tool kit; hymenopteran-specific genes including diverse venoms; lateral gene transfers among Pox viruses, Wolbachia, and Nasonia; and the rapid evolution of genes involved in nuclear-mitochondrial interactions that are implicated in speciation. Newly developed genome resources advance Nasonia for genetic research, accelerate mapping and cloning of quantitative trait loci, and will ultimately provide tools and knowledge for further increasing the utility of parasitoids as pest insect-control agents.}, } @article {pmid20071985, year = {2010}, author = {Murdoch, ME}, title = {Onchodermatitis.}, journal = {Current opinion in infectious diseases}, volume = {23}, number = {2}, pages = {124-131}, doi = {10.1097/QCO.0b013e328336a256}, pmid = {20071985}, issn = {1473-6527}, mesh = {Africa ; Animals ; Anti-Bacterial Agents/therapeutic use ; Dermatitis/*parasitology ; Doxycycline/therapeutic use ; Filaricides/therapeutic use ; Humans ; Insect Control ; Ivermectin/therapeutic use ; Latin America ; Macrolides/therapeutic use ; Onchocerciasis/diagnosis/*drug therapy/*pathology ; Skin Diseases, Parasitic/diagnosis/*drug therapy/*pathology ; }, abstract = {PURPOSE OF REVIEW: This review is timely because awareness of the burden of disease from onchodermatitis has increased significantly over recent years. Recent progress in the field is reviewed with emphasis on publications within the past 2 years.

RECENT FINDINGS: Advances have been made in understanding immunopathogenesis and in diagnosis and treatment. The World Bank/WHO African Programme for Onchocerciasis Control (APOC), which uses annual community-directed treatment with ivermectin (CDTI) via the Mectizan Donation Programme, now covers 19 African countries. Development of ivermectin resistance is a concern. Unlike ivermectin, which is a microfilaricide, doxycycline, which targets Wolbachia endosymbiotic bacteria, sterilizes adult female worms and has a macrofilaricidal effect. Moxidectin, which sterilizes or kills adult worms has started a phase III trial with ivermectin. Additional primary healthcare interventions have been successfully integrated with CTDI. In Latin America, transmission has been interrupted in half of the original endemic foci and Colombia is the first nation to have achieved countrywide interruption of transmission. The first report of elimination using ivermectin in an African setting is a milestone. Two African foci using vector control plus CDTI have reported vector elimination.

SUMMARY: Results of the longer-term impact of large-scale ivermectin distribution by the APOC are awaited. Research is needed into new drug targets within Wolbachia's metabolic pathways. Elimination of transmission of disease is on the horizon but more research is needed on when and where ivermectin treatment can be stopped.}, } @article {pmid20064373, year = {2009}, author = {Moreira, LA and Iturbe-Ormaetxe, I and Jeffery, JA and Lu, G and Pyke, AT and Hedges, LM and Rocha, BC and Hall-Mendelin, S and Day, A and Riegler, M and Hugo, LE and Johnson, KN and Kay, BH and McGraw, EA and van den Hurk, AF and Ryan, PA and O'Neill, SL}, title = {A Wolbachia symbiont in Aedes aegypti limits infection with dengue, Chikungunya, and Plasmodium.}, journal = {Cell}, volume = {139}, number = {7}, pages = {1268-1278}, doi = {10.1016/j.cell.2009.11.042}, pmid = {20064373}, issn = {1097-4172}, mesh = {Aedes/*microbiology/parasitology/physiology/virology ; Animals ; Chikungunya virus/*physiology ; Dengue Virus/*physiology ; Host-Parasite Interactions ; Plasmodium gallinaceum/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacterial symbionts that are estimated to infect more than 60% of all insect species. While Wolbachia is commonly found in many mosquitoes it is absent from the species that are considered to be of major importance for the transmission of human pathogens. The successful introduction of a life-shortening strain of Wolbachia into the dengue vector Aedes aegypti that halves adult lifespan has recently been reported. Here we show that this same Wolbachia infection also directly inhibits the ability of a range of pathogens to infect this mosquito species. The effect is Wolbachia strain specific and relates to Wolbachia priming of the mosquito innate immune system and potentially competition for limiting cellular resources required for pathogen replication. We suggest that this Wolbachia-mediated pathogen interference may work synergistically with the life-shortening strategy proposed previously to provide a powerful approach for the control of insect transmitted diseases.}, } @article {pmid20049511, year = {2010}, author = {Robe, LJ and Cordeiro, J and Loreto, EL and Valente, VL}, title = {Taxonomic boundaries, phylogenetic relationships and biogeography of the Drosophila willistoni subgroup (Diptera: Drosophilidae).}, journal = {Genetica}, volume = {138}, number = {6}, pages = {601-617}, pmid = {20049511}, issn = {1573-6857}, mesh = {Animals ; Biological Evolution ; Drosophila/*classification/*genetics/microbiology ; Phylogeny ; Wolbachia ; }, abstract = {The Drosophila willistoni subgroup represents a complex with varying taxonomic levels. It encompasses D. willistoni and its five sibling species: D. equinoxialis, D. insularis, D. paulistorum, D. pavlovskiana and D. tropicalis. Of these, D. equinoxialis, D. tropicalis and D. willistoni present differentiation at subspecific level, whereas D. paulistorum represents a superspecies, formed by six semispecies. Despite this taxonomic and evolutionary complexity, many of these semi and subspecific taxa have not yet had their phylogenetic status tested in an explicitly molecular study. Aiming to contribute to the understanding of the evolution of this challenging group, we analyzed nucleotide sequences from two mitochondrial and four nuclear datasets, both individually and simultaneously, through different phylogenetic methods. High levels of incongruence were detected among partitions, especially concerning the mitochondrial sequences. As this incongruence was found to be statistically significant and robust to the use of different models and approaches, and basically restricted to mitochondrial loci, we suggest that it may stem mainly from hybridization-mediated asymmetrical introgression. Despite this, our nuclear data finally led to a phylogenetic hypothesis which further refines several aspects related to the willistoni subgroup phylogeny. In this respect, D. insularis, D. tropicalis, D. willistoni and D. equinoxialis successively branched off from the willistoni subgroup main stem, which recently subdivided to produce D. paulistorum and D. pavlovskiana. As regards the semispecies evolution, we found evidence of a recent diversification, which highly influenced the obtained results due to the associated small levels of genetic differentiation, further worsened by the possibly associated incompletely sorted ancestral polymorphisms and by the possibility of introgression. This study also raises the question of whether these semispecies are monophyletic at all. This reasoning is particularly interesting when one considers that similar levels of reproductive isolation could be attained through infection with different Wolbachia strains.}, } @article {pmid20040002, year = {2010}, author = {Hornett, EA and Engelstädter, J and Hurst, GD}, title = {Hidden cytoplasmic incompatibility alters the dynamics of male-killer/host interactions.}, journal = {Journal of evolutionary biology}, volume = {23}, number = {3}, pages = {479-487}, doi = {10.1111/j.1420-9101.2009.01872.x}, pmid = {20040002}, issn = {1420-9101}, mesh = {Animals ; Arthropods/*microbiology ; Biological Evolution ; Female ; Male ; *Models, Biological ; Wolbachia/*physiology ; }, abstract = {Wolbachia manipulate the reproduction of their arthropod hosts in a variety of ways. Recent work has demonstrated that these bacteria may combine phenotypes - possessing a 'male killing' phenotype and, where males survive, induce cytoplasmic incompatibility (CI). We here develop a mathematical model to investigate the extent to which 'hidden' CI may affect the evolutionary dynamics of host genes that suppress male-killing activity. We observed that for high prevalence infections, CI drives both suppressor and bacterium to higher frequency, such that the strain appears to solely exhibit CI. In contrast, for low prevalence infections, CI impedes suppressor invasion as surviving infected males are incompatible with the majority of females in the population. Our results demonstrate that 'hidden' phenotypes as well as observable ones can impact on the dynamics of the interaction, and knowledge of these is therefore required to predict when suppressor genes will invade, and the consequences of their invasion.}, } @article {pmid20036185, year = {2010}, author = {Saridaki, A and Bourtzis, K}, title = {Wolbachia: more than just a bug in insects genitals.}, journal = {Current opinion in microbiology}, volume = {13}, number = {1}, pages = {67-72}, doi = {10.1016/j.mib.2009.11.005}, pmid = {20036185}, issn = {1879-0364}, mesh = {Animals ; Culex/*microbiology ; Eukaryotic Cells/*microbiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Research on the intracellular bacterial symbiont Wolbachia has grown on many levels, providing interesting insights on various aspects of the microbe's biology. Although data from fully sequenced genomes of different Wolbachia strains and from experimental studies of host-microbe interactions continue to arise, most of the molecular mechanisms employed by Wolbachia to manipulate the host cytoplasmic machinery and to ensure vertical transmission are yet to be discovered. Apart from the well-established role of Wolbachia in triggering reproductive alterations, a new fascinating aspect is emerging, related to the ecological benefits that the symbiont provides to the host. The mutualistic relationship of Wolbachia strains with disease vectors remains among the top research priorities with new insights having an impact on putative anti-filarial strategies.}, } @article {pmid20034492, year = {2010}, author = {Tompkins, JB and Stitt, LE and Ardelli, BF}, title = {Brugia malayi: in vitro effects of ivermectin and moxidectin on adults and microfilariae.}, journal = {Experimental parasitology}, volume = {124}, number = {4}, pages = {394-402}, doi = {10.1016/j.exppara.2009.12.003}, pmid = {20034492}, issn = {1090-2449}, mesh = {Analysis of Variance ; Animals ; Antinematodal Agents/*pharmacology ; Antiparasitic Agents/*pharmacology ; Brugia malayi/*drug effects/microbiology/physiology ; Female ; Fertility/drug effects ; Gerbillinae ; Ivermectin/*pharmacology ; Macrolides/pharmacology ; Male ; Microfilariae/drug effects/microbiology/physiology ; Movement/drug effects ; Polymerase Chain Reaction ; RNA, Bacterial/analysis ; Random Allocation ; Time Factors ; Wolbachia/genetics/isolation & purification ; }, abstract = {The effect of ivermectin and moxidectin on the motility of Brugia malayi adults and microfilariae and on the fertility of B. malayi females was examined. Motility was reduced in adults after exposure to both drugs and worms were non-motile and dead within eight days. The motility of microfilariae was significantly reduced at all drug concentrations and ceased at concentrations of 2500 and 5000mug/mL. The motility of microfilariae released by females was reduced after exposure to both drugs, however ivermectin had a greater effect at concentrations between 170 and 5000mug/mL. Both drugs reduced the number of microfilariae released by females and within four days their release was inhibited. The presence of the bacterial endosymbiont Wolbachia was examined in adults and microfilariae after exposure to increasing concentrations of ivermectin and moxidectin. A decrease in wsp expression was correlated with increasing drug concentration.}, } @article {pmid20021758, year = {2009}, author = {Kawai, S and Matsumoto, Y and Gotoh, T and Noda, H}, title = {Transinfection of Wolbachia in planthoppers: nymphal injection of cultured Wolbachia and infection dynamics.}, journal = {Environmental entomology}, volume = {38}, number = {6}, pages = {1626-1633}, doi = {10.1603/022.038.0615}, pmid = {20021758}, issn = {1938-2936}, mesh = {Animals ; Female ; Hemiptera/*microbiology ; *Host-Pathogen Interactions ; Male ; Microinjections ; Nymph/microbiology ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia species are intracellular symbionts that cause reproductive alterations in arthropods. Transinfection experiments have been performed in many arthropod species to elucidate the interaction between Wolbachia and a new host. To ease transinfection of this bacterium to new arthropod hosts, we introduced two techniques: nymphal injection instead of embryonic injection and the use of a cultured source of Wolbachia instead of direct transfer from donors to recipients. Wolbachia in the small brown planthopper Laodelphax striatellus was cultivated in a cell line and injected into the nymphal body cavity of the brown planthopper Nilaparvata lugens together with the cells. By using these techniques, two transinfected planthopper lines were obtained. In one line, Wolbachia disappeared after several generations; in the other line, Wolbachia was retained for >7 yr. Infection rates in this latter transinfected line were approximately 80% in early generations after transinjection but decreased to <10% through 40-60 generations. Subsequent selection for Wolbachia-infected females in this line did not increase the infection rate as a temporary effect. Thus, this transinfected line of N. lugens showed cytoplasmic incompatibility, although the incompatibility level was lower than in L. striatellus, the original host. The method of transinfection presented herein is useful for transmitting intracellular symbionts between small arthropod hosts.}, } @article {pmid20016848, year = {2009}, author = {Moreira, LA and Saig, E and Turley, AP and Ribeiro, JM and O'Neill, SL and McGraw, EA}, title = {Human probing behavior of Aedes aegypti when infected with a life-shortening strain of Wolbachia.}, journal = {PLoS neglected tropical diseases}, volume = {3}, number = {12}, pages = {e568}, pmid = {20016848}, issn = {1935-2735}, mesh = {Aedes/enzymology/*microbiology/*physiology ; Animals ; Apyrase/metabolism ; Feeding Behavior ; Female ; Humans ; Insect Proteins/metabolism ; Insect Vectors/enzymology/microbiology/physiology ; Time Factors ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {BACKGROUND: Mosquitoes are vectors of many serious pathogens in tropical and sub-tropical countries. Current control strategies almost entirely rely upon insecticides, which increasingly face the problems of high cost, increasing mosquito resistance and negative effects on non-target organisms. Alternative strategies include the proposed use of inherited life-shortening agents, such as the Wolbachia bacterium. By shortening mosquito vector lifespan, Wolbachia could potentially reduce the vectorial capacity of mosquito populations. We have recently been able to stably transinfect Aedes aegypti mosquitoes with the life-shortening Wolbachia strain wMelPop, and are assessing various aspects of its interaction with the mosquito host to determine its likely impact on pathogen transmission as well as its potential ability to invade A. aegypti populations.

Here we have examined the probing behavior of Wolbachia-infected mosquitoes in an attempt to understand both the broader impact of Wolbachia infection on mosquito biology and, in particular, vectorial capacity. The probing behavior of wMelPop-infected mosquitoes at four adult ages was examined and compared to uninfected controls during video-recorded feeding trials on a human hand. Wolbachia-positive insects, from 15 days of age, showed a drastic increase in the time spent pre-probing and probing relative to uninfected controls. Two other important features for blood feeding, saliva volume and apyrase content of saliva, were also studied.

CONCLUSIONS/SIGNIFICANCE: As A. aegypti infected with wMelPop age, they show increasing difficulty in completing the process of blood feeding effectively and efficiently. Wolbachia-infected mosquitoes on average produced smaller volumes of saliva that still contained the same amount of apyrase activity as uninfected mosquitoes. These effects on blood feeding behavior may reduce vectorial capacity and point to underlying physiological changes in Wolbachia-infected mosquitoes.}, } @article {pmid20005876, year = {2010}, author = {Crainey, JL and Wilson, MD and Post, RJ}, title = {Phylogenetically distinct Wolbachia gene and pseudogene sequences obtained from the African onchocerciasis vector Simulium squamosum.}, journal = {International journal for parasitology}, volume = {40}, number = {5}, pages = {569-578}, doi = {10.1016/j.ijpara.2009.10.017}, pmid = {20005876}, issn = {1879-0135}, support = {G0600015/MRC_/Medical Research Council/United Kingdom ; 77615/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Bacterial Proteins/*genetics ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; *Disease Vectors ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology ; Simuliidae/*microbiology ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Wolbachia are intracellular bacteria mostly found in a diverse range of arthropods and filarial nematodes. They have been classified into seven distinct 'supergroups' and other lineages on the basis of molecular phylogenetics. The arthropod-infecting Wolbachia are usually regarded as reproductive parasites because they manipulate their host species' sexing system to enhance their own spread, and this has led to their investigation as potential agents of genetic control in medical entomology. We report 12 partial Wolbachia gene sequences from: aspC, aspS, dnaA, fbpA, ftsZ, GroEL, hcpA, IDA, rpoB, rpe, TopI and wsp as well as a single ftsZ pseudogene sequence, which have all been PCR-amplified from Simulium squamosum (Diptera: Simuliidae). To our knowledge this is the first such report from Simuliidae. Uninterrupted open-reading frame sequences were obtained from all 12 genes, covering approximately 6.2kb of unique DNA sequence. Phylogenetic analyses with the different coding genes gave consistent results suggesting that the Wolbachia sequences obtained here do not derive from any of the known Wolbachia supergroups or lineages. Consistent with a unique genetic status for the S. squamosumWolbachia, the hypervariable regions of the Wolbachia-specific wsp gene were distinct from all previous records in both sequence and length. As well as potential implications for newly emerging Wolbachia-based disease control methods, the results may be relevant to some problems experienced in the laboratory colonisation of Simulium damnosum sensu lato and why it is such a diverse species complex.}, } @article {pmid20002580, year = {2010}, author = {Jaenike, J and Stahlhut, JK and Boelio, LM and Unckless, RL}, title = {Association between Wolbachia and Spiroplasma within Drosophila neotestacea: an emerging symbiotic mutualism?.}, journal = {Molecular ecology}, volume = {19}, number = {2}, pages = {414-425}, doi = {10.1111/j.1365-294X.2009.04448.x}, pmid = {20002580}, issn = {1365-294X}, mesh = {Animals ; Cytoplasm/genetics ; DNA, Bacterial/genetics ; DNA, Mitochondrial/genetics ; Drosophila/genetics/*microbiology ; Evolution, Molecular ; Female ; Haplotypes ; Male ; *Phylogeny ; Spiroplasma/*genetics ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Interspecific mutualism can evolve when specific lineages of different species tend to be associated with each other from one generation to the next. Different maternally transmitted endosymbionts occurring within the same cytoplasmic lineage fulfil this requirement. Drosophila neotestacea is infected with maternally transmitted Wolbachia and Spiroplasma, which are cotransmitted at high frequency in natural populations. Molecular phylogenetic evidence indicates that both endosymbionts have been present in D. neotestacea for considerable evolutionary periods. Thus, conditions are suitable for the evolution of mutualism between them. In support of this possibility, there is a significant positive association between Wolbachia and Spiroplasma infection in many samples of D. neotestacea from natural populations. Theoretically, such a positive association can result from either mutualism between these endosymbionts or recent spread. Collections from present-day populations suggest that recent spread and mutualism have both operated to generate the positive association between Wolbachia and Spiroplasma. If selection acts on the combination of these two endosymbionts, they may be in the early stages of evolution of a more complex, cooperative association.}, } @article {pmid19956588, year = {2009}, author = {Jeffery, JA and Thi Yen, N and Nam, VS and Nghia, le T and Hoffmann, AA and Kay, BH and Ryan, PA}, title = {Characterizing the Aedes aegypti population in a Vietnamese village in preparation for a Wolbachia-based mosquito control strategy to eliminate dengue.}, journal = {PLoS neglected tropical diseases}, volume = {3}, number = {11}, pages = {e552}, pmid = {19956588}, issn = {1935-2735}, mesh = {Adolescent ; Adult ; Aedes/*microbiology/physiology ; Animals ; Child ; Child, Preschool ; Dengue/epidemiology/*prevention & control ; Female ; Housing ; Humans ; Insect Vectors/*microbiology/physiology ; Male ; Middle Aged ; *Mosquito Control ; *Pest Control, Biological ; Population Density ; Population Dynamics ; Rural Health ; *Rural Population ; Vietnam/epidemiology ; Wolbachia/*physiology ; Young Adult ; }, abstract = {BACKGROUND: A life-shortening strain of the obligate intracellular bacteria Wolbachia, called wMelPop, is seen as a promising new tool for the control of Aedes aegypti. However, developing a vector control strategy based on the release of mosquitoes transinfected with wMelPop requires detailed knowledge of the demographics of the target population.

In Tri Nguyen village (611 households) on Hon Mieu Island in central Vietnam, we conducted nine quantitative entomologic surveys over 14 months to determine if Ae. aegypti populations were spatially and temporally homogenous, and to estimate population size. There was no obvious relationship between mosquito (larval, pupal or adult) abundance and temperature and rainfall, and no area of the village supported consistently high numbers of mosquitoes. In almost all surveys, key premises produced high numbers of Ae. aegypti. However, these premises were not consistent between surveys. For an intervention based on a single release of wMelPop-infected Ae. aegypti, release ratios of infected to uninfected adult mosquitoes of all age classes are estimated to be 1.8-6.7ratio1 for gravid females (and similarly aged males) or teneral adults, respectively. We calculated that adult female mosquito abundance in Tri Nguyen village could range from 1.1 to 43.3 individuals of all age classes per house. Thus, an intervention could require the release of 2-78 wMelPop-infected gravid females and similarly aged males per house, or 7-290 infected teneral female and male mosquitoes per house.

CONCLUSIONS/SIGNIFICANCE: Given the variability we encountered, this study highlights the importance of multiple entomologic surveys when evaluating the spatial structure of a vector population or estimating population size. If a single release of wMelPop-infected Ae. aegypti were to occur when wild Ae. aegypti abundance was at its maximum, a preintervention control program would be necessary to ensure that there was no net increase in mosquito numbers. However, because of the short-term temporal heterogeneity, the inconsistent spatial structure and the impact of transient key premises that we observed, the feasibility of multiple releases of smaller numbers of mosquitoes also needs to be considered. In either case, fewer wMelPop-infected mosquitoes would then need to be released, which will likely be more acceptable to householders.}, } @article {pmid19954456, year = {2009}, author = {Shiny, C and Krushna, NS and Archana, B and Farzana, B and Narayanan, RB}, title = {Serum antibody responses to Wolbachia surface protein in patients with human lymphatic filariasis.}, journal = {Microbiology and immunology}, volume = {53}, number = {12}, pages = {685-693}, doi = {10.1111/j.1348-0421.2009.00172.x}, pmid = {19954456}, issn = {0385-5600}, mesh = {Amino Acid Sequence ; Animals ; Antibodies/*blood ; B-Lymphocytes/immunology ; Bacterial Proteins/genetics/immunology ; Brugia malayi/*microbiology ; Cloning, Molecular ; DNA, Protozoan/genetics ; Elephantiasis, Filarial/blood/immunology/microbiology ; Epitopes/analysis/chemistry/immunology ; HLA-D Antigens/immunology ; Helminth Proteins/genetics ; Humans ; Immunoglobulin E/blood ; Immunoglobulin G/blood ; Membrane Proteins/genetics/immunology ; Polymerase Chain Reaction ; T-Lymphocytes/immunology ; Wolbachia/*immunology ; }, abstract = {Wolbachia surface protein (WSP), which is the most abundantly expressed protein of Wolbachia from the human filarial parasite Brugia malayi, was chosen for the present study. B-cell epitope prediction of the WSP protein sequence indicates a high antigenicity, surface probability and hydrophilicity by DNA STAR software analysis. ProPred analysis suggests the presence of HLA class II binding regions in the WSP protein that contribute to T-cell responses and isotype reactivity. In order to validate these findings, the gene coding for endosymbiont WSP was PCR-amplified from the genomic DNA of the human filarial parasite Brugia malayi and cloned in T-7 expression vector pRSET-A. Western blot and ELISA at the total IgG level with recombiant WSP indicated a significantly elevated reactivity in CP compared to MF, EN and NEN individuals. Isotype ELISA also suggested an elevated reactivity in CP patients at the IgG1 level. In contrast, WSP-specific IgG4 levels were found to be elevated in MF patients compared to CP and EN. Besides this, WSP-specific IgE levels indicated an elevated reactivity in CP and MF patients compared to normals. Observations from ELISA supported the in silico predictions that indicate the presence of B- and T-cell epitopes. Hence, a combinatorial approach of in silico predictions and wet-lab studies provides interesting insights into the role of Wolbachia proteins in filarial pathogenesis.}, } @article {pmid19951376, year = {2010}, author = {Voronin, D and Tran-Van, V and Potier, P and Mavingui, P}, title = {Transinfection and growth discrepancy of Drosophila Wolbachia strain wMel in cell lines of the mosquito Aedes albopictus.}, journal = {Journal of applied microbiology}, volume = {108}, number = {6}, pages = {2133-2141}, doi = {10.1111/j.1365-2672.2009.04621.x}, pmid = {19951376}, issn = {1365-2672}, mesh = {Aedes/cytology/genetics/*microbiology ; Animals ; Cell Culture Techniques ; Cell Line ; Cytoplasm/microbiology ; In Situ Hybridization, Fluorescence ; Polymerase Chain Reaction ; Wolbachia/*growth & development/isolation & purification ; }, abstract = {AIM: The Wolbachia strain wMel can protect Drosophila melanogaster against pathogenic RNA viruses. To analyse the potential of this inhibitory effect against arboviruses vectorized by these mosquitoes, we here first transinfected the Aedes albopictus Aa23 and C6/36 cell lines with the Wolbachia strain wMel and then monitored their infection dynamics.

METHODS AND RESULTS: Wolbachia strain wMel was transferred into A. albopictus Aa23 and C6/36 cell lines using the shell vial technique. The presence of the bacterium in the transinfected cells was monitored by quantitative PCR and fluorescence in situ hybridization. Bacteria could be detected in the cytoplasm of both the Aa23 and C6/36 cell lines. However, the dynamics and stability of the bacterial infection differed depending on the initial cell background. The Aa23 cell line, which had been treated with a tetracycline antibiotic 2 years previously to eliminate its natural Wolbachia wAlbB-infecting strain, lost the introduced Wolbachia wMel strain after 12 passages postinfection. In contrast, the C6/36 cell line, which had originally been aposymbiotic, displayed a stable infection with Wolbachia wMel. The bacterial density in C6/36 was greater than that of the A. albopictus RML12 cell line from which the wMel strain had originated.

CONCLUSIONS: Transient or persistent transinfection of A. albopictus Aa23 and C6/36 cell lines with Wolbachia wMel strain was achieved. The results indicate the influence of the genetic background of mosquito cells in maintaining Wolbachia originating from a distant dipteral host.

The cell model built here can now be used to investigate the viral inhibitory effect of the Wolbachia wMel strain against arboviruses such as dengue and chikungunya, which are transmitted by the mosquito A. albopictus.}, } @article {pmid19943957, year = {2009}, author = {Holman, AG and Davis, PJ and Foster, JM and Carlow, CK and Kumar, S}, title = {Computational prediction of essential genes in an unculturable endosymbiotic bacterium, Wolbachia of Brugia malayi.}, journal = {BMC microbiology}, volume = {9}, number = {}, pages = {243}, pmid = {19943957}, issn = {1471-2180}, mesh = {Animals ; Brugia malayi/*microbiology ; *Computational Biology ; Conserved Sequence/genetics ; Drug Design ; Genes, Bacterial/*genetics ; Genome, Bacterial ; Rickettsiaceae/genetics ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Wolbachia (wBm) is an obligate endosymbiotic bacterium of Brugia malayi, a parasitic filarial nematode of humans and one of the causative agents of lymphatic filariasis. There is a pressing need for new drugs against filarial parasites, such as B. malayi. As wBm is required for B. malayi development and fertility, targeting wBm is a promising approach. However, the lifecycle of neither B. malayi nor wBm can be maintained in vitro. To facilitate selection of potential drug targets we computationally ranked the wBm genome based on confidence that a particular gene is essential for the survival of the bacterium.

RESULTS: wBm protein sequences were aligned using BLAST to the Database of Essential Genes (DEG) version 5.2, a collection of 5,260 experimentally identified essential genes in 15 bacterial strains. A confidence score, the Multiple Hit Score (MHS), was developed to predict each wBm gene's essentiality based on the top alignments to essential genes in each bacterial strain. This method was validated using a jackknife methodology to test the ability to recover known essential genes in a control genome. A second estimation of essentiality, the Gene Conservation Score (GCS), was calculated on the basis of phyletic conservation of genes across Wolbachia's parent order Rickettsiales. Clusters of orthologous genes were predicted within the 27 currently available complete genomes. Druggability of wBm proteins was predicted by alignment to a database of protein targets of known compounds.

CONCLUSION: Ranking wBm genes by either MHS or GCS predicts and prioritizes potentially essential genes. Comparison of the MHS to GCS produces quadrants representing four types of predictions: those with high confidence of essentiality by both methods (245 genes), those highly conserved across Rickettsiales (299 genes), those similar to distant essential genes (8 genes), and those with low confidence of essentiality (253 genes). These data facilitate selection of wBm genes for entry into drug design pipelines.}, } @article {pmid19934219, year = {2009}, author = {Albertson, R and Casper-Lindley, C and Cao, J and Tram, U and Sullivan, W}, title = {Symmetric and asymmetric mitotic segregation patterns influence Wolbachia distribution in host somatic tissue.}, journal = {Journal of cell science}, volume = {122}, number = {Pt 24}, pages = {4570-4583}, pmid = {19934219}, issn = {1477-9137}, support = {5R01GM046409/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Brain/microbiology ; Cell Division ; Centrosome/microbiology/physiology ; Drosophila melanogaster/embryology/*microbiology/*physiology ; Microtubules/microbiology/physiology ; *Mitosis ; Organ Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited bacterial endosymbionts that occupy many but not all tissues of adult insects. During the initial mitotic divisions in Drosophila embryogenesis, Wolbachia exhibit a symmetric pattern of segregation. Wolbachia undergo microtubule-dependent and cell-cycle-regulated movement between centrosomes. Symmetric segregation occurs during late anaphase when Wolbachia cluster around duplicated and separating centrosomes. This centrosome association is microtubule-dependent and promotes an even Wolbachia distribution throughout the host embryo. By contrast, during the later embryonic and larval neuroblast divisions, Wolbachia segregate asymmetrically with the apical self-renewing neuroblast. During these polarized asymmetric neuroblast divisions, Wolbachia colocalize with the apical centrosome and apically localized Par complex. This localization depends on microtubules, but not the cortical actin-based cytoskeleton. We also found that Wolbachia concentrate in specific regions of the adult brain, which might be a direct consequence of the asymmetric Wolbachia segregation in the earlier neuroblast divisions. Finally, we demonstrate that the fidelity of asymmetric segregation to the self-renewing neuroblast is lower in the virulent Popcorn strain of Wolbachia.}, } @article {pmid19916630, year = {2010}, author = {Michalski, ML and Bain, O and Fischer, K and Fischer, PU and Kumar, S and Foster, JM}, title = {Identification and phylogenetic analysis of Dirofilaria ursi (Nematoda: Filarioidea) from Wisconsin black bears (Ursus americanus) and its Wolbachia endosymbiont.}, journal = {The Journal of parasitology}, volume = {96}, number = {2}, pages = {412-419}, doi = {10.1645/GE-2208.1}, pmid = {19916630}, issn = {1937-2345}, support = {R15 AI067295/AI/NIAID NIH HHS/United States ; 1R15AI067295-01A/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; DNA, Bacterial/chemistry ; DNA, Helminth/chemistry ; DNA, Ribosomal/chemistry ; Dirofilaria/anatomy & histology/*classification/genetics/microbiology ; Dirofilariasis/epidemiology/*parasitology ; Dogs ; Female ; Immunohistochemistry/veterinary ; Male ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/veterinary ; Prevalence ; RNA, Ribosomal, 5S/genetics ; Sequence Alignment/veterinary ; *Symbiosis ; Ursidae/*parasitology ; Wisconsin/epidemiology ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Dirofilaria ursi is a filarial nematode of American black bears (Ursus americanus Pallas, 1780) that is vectored by black flies (Simuliidae) in many parts of the United States. In northwestern Wisconsin, the prevalence of microfilaremic bears during the fall hunting season was 21% (n = 47). Unsheathed blood microfilariae from Wisconsin bears possess characters consistent with the original description of D. ursi, as do adult worms observed histologically and grossly. Immunohistochemistry was used to identify the Wolbachia endosymbiont in the hypodermis and lateral cords of an adult female D. ursi. Amplification of wsp, gatB, coxA, fbpA, and ftsZ bacterial sequences from parasite DNA confirmed the presence of Wolbachia, and molecular phylogenetic analysis of the Wolbachia ftsZ gene groups the endosymbiont with Wolbachia from D. immitis and D. repens. Phylogenetic analysis of D. ursi 5s rDNA sequence confirms the morphological observations grouping this parasite as a member of Dirofilaria, and within the Dirofilaria - Onchocerca clade of filarial nematodes. This is the first report of Wolbachia characterization and molecular phylogeny information for D. ursi.}, } @article {pmid19912450, year = {2010}, author = {Duplouy, A and Hurst, GD and O'Neill, SL and Charlat, S}, title = {Rapid spread of male-killing Wolbachia in the butterfly Hypolimnas bolina.}, journal = {Journal of evolutionary biology}, volume = {23}, number = {1}, pages = {231-235}, doi = {10.1111/j.1420-9101.2009.01891.x}, pmid = {19912450}, issn = {1420-9101}, mesh = {Animals ; Butterflies/genetics/*microbiology ; DNA, Bacterial/chemistry ; DNA, Mitochondrial/chemistry ; Female ; Male ; Sequence Analysis, DNA ; Wolbachia/genetics/growth & development/*physiology ; }, abstract = {Reproductive parasites such as Wolbachia can spread through uninfected host populations by increasing the relative fitness of the infected maternal lineage. However, empirical estimates of how fast this process occurs are limited. Here we use nucleotide sequences of male-killing Wolbachia bacteria and co-inherited mitochondria to address this issue in the island butterfly Hypolimnas bolina. We show that infected specimens scattered throughout the species range harbour the same Wolbachia and mitochondrial DNA as inferred from 6337 bp of the bacterial genome and 2985 bp of the mitochondrial genome, suggesting this strain of Wolbachia has spread across the South Pacific Islands at most 3000 years ago, and probably much more recently.}, } @article {pmid19911047, year = {2009}, author = {Osborne, SE and Leong, YS and O'Neill, SL and Johnson, KN}, title = {Variation in antiviral protection mediated by different Wolbachia strains in Drosophila simulans.}, journal = {PLoS pathogens}, volume = {5}, number = {11}, pages = {e1000656}, pmid = {19911047}, issn = {1553-7374}, mesh = {Animals ; *Drosophila/immunology/microbiology/virology ; Host-Pathogen Interactions/*immunology ; Insect Viruses/*pathogenicity ; Kaplan-Meier Estimate ; Symbiosis/*immunology ; Viral Load ; Virus Diseases/immunology/microbiology/virology ; Wolbachia/genetics/*physiology ; }, abstract = {Drosophila C virus (DCV) is a natural pathogen of Drosophila and a useful model for studying antiviral defences. The Drosophila host is also commonly infected with the widespread endosymbiotic bacteria Wolbachia pipientis. When DCV coinfects Wolbachia-infected D. melanogaster, virus particles accumulate more slowly and virus induced mortality is substantially delayed. Considering that Wolbachia is estimated to infect up to two-thirds of all insect species, the observed protective effects of Wolbachia may extend to a range of both beneficial and pest insects, including insects that vector important viral diseases of humans, animals and plants. Currently, Wolbachia-mediated antiviral protection has only been described from a limited number of very closely related strains that infect D. melanogaster. We used D. simulans and its naturally occurring Wolbachia infections to test the generality of the Wolbachia-mediated antiviral protection. We generated paired D. simulans lines either uninfected or infected with five different Wolbachia strains. Each paired fly line was challenged with DCV and Flock House virus. Significant antiviral protection was seen for some but not all of the Wolbachia strain-fly line combinations tested. In some cases, protection from virus-induced mortality was associated with a delay in virus accumulation, but some Wolbachia-infected flies were tolerant to high titres of DCV. The Wolbachia strains that did protect occurred at comparatively high density within the flies and were most closely related to the D. melanogaster Wolbachia strain wMel. These results indicate that Wolbachia-mediated antiviral protection is not ubiquitous, a finding that is important for understanding the distribution of Wolbachia and virus in natural insect populations.}, } @article {pmid19906794, year = {2010}, author = {Chafee, ME and Funk, DJ and Harrison, RG and Bordenstein, SR}, title = {Lateral phage transfer in obligate intracellular bacteria (wolbachia): verification from natural populations.}, journal = {Molecular biology and evolution}, volume = {27}, number = {3}, pages = {501-505}, pmid = {19906794}, issn = {1537-1719}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/*genetics ; Bayes Theorem ; Coleoptera/genetics/*microbiology/virology ; Evolution, Molecular ; *Gene Transfer, Horizontal ; Gryllidae/genetics/*microbiology/virology ; Interspersed Repetitive Sequences ; Phylogeny ; Wolbachia/*genetics/virology ; }, abstract = {Lateral transfer of mobile DNA is a hallmark of bacteria with a free-living replicative stage; however, its significance in obligate intracellular bacteria and other heritable endosymbionts remains controversial. Comparative sequence analyses from laboratory stocks infected with Wolbachia pipientis provide some of the most compelling evidence that bacteriophage WO-B transfers laterally between infections of the same insect host. Lateral transfer between coinfections, however, has been evaluated neither in natural populations nor between closely related Wolbachia strains. Here, we analyze bacterial and phage genes from two pairs of natural sympatric field isolates, of Gryllus pennsylvanicus field crickets and of Neochlamisus bebbianae leaf beetles, to demonstrate WO-B transfers between supergroup B Wolbachia. N. bebbianae revealed the highest number of phage haplotypes yet recorded, hinting that lab lines could underestimate phage haplotype variation and lateral transfer. Finally, using the approximate age of insect host species as the maximum available time for phage transfer between host-associated bacteria, we very conservatively estimate phage WO-B transfer to occur at least once every 0-5.4 My within a host species. Increasing discoveries of mobile elements, intragenic recombination, and bacterial coinfections in host-switching obligate intracellular bacteria specify that mobile element transfer is common in these species.}, } @article {pmid19891414, year = {2009}, author = {Vaĭsman, NIa and Ilinskiĭ, IuIu and Golubovskiĭ, MD}, title = {[Population genetic analysis of Drosophila melanogaster longevity: similar effects of endosymbiont Wolbachia and tumor suppressor lgl under conditions of temperature stress].}, journal = {Zhurnal obshchei biologii}, volume = {70}, number = {5}, pages = {438-447}, pmid = {19891414}, issn = {0044-4596}, mesh = {Animals ; Drosophila Proteins/*genetics ; Drosophila melanogaster/*genetics/growth & development/microbiology ; *Genetics, Population ; Longevity/*genetics ; Stress, Physiological/*genetics ; *Symbiosis ; Temperature ; Tumor Suppressor Proteins/*genetics ; Wolbachia/*growth & development ; }, abstract = {Before, it has been shown that keeping of Drosophila wild populations under temperature stress leads to prevalence of deletion alleles of tumor suppressor lgl, and higher survival of heterozygous lgl(-)/+ females descendants (haploadaptivity). On the other hand, recent investigations have demonstrated that an additional dose of lgl gene in transgenic males has inadaptive effect--cytoplasmic incompatibility, caused by the gene, is similar to the effect caused by the widespread endosymbiont Wolbachia. Both triple dose of lgl and Wolbachia infection invoke hyperexpression of some cytoskeleton proteins which leads to sperm defects and sterility of hybrid zygotes. Proceeding from this similarity, we studied the Wolbachia effect on Drosophila longevity (LG) depending on the bacteria strain, host genotype and sex, and maintenance temperature. With use of tetracycline, six Wolbachia-free lines were produced, analogical to the infected wild ones. After analyzing LG under optimal temperature conditions (25 degrees C), the lines were subdivided into three genotypic groups, differing by the LG, mortality dynamics, and type of sexual dimorphism. Under conditions of temperature stress (16 degrees C and 29 degrees C) the lines ranks with respect to LG remained as they were, though the distinctions between them were smoothed out. In other words, the effect of the genes regulating LG parameters depends on maintenance conditions. The research also revealed sharp genetic distinctions between LG parameters of two standard laboratory lines Canton and Oregon that were taken as the control. These distinctions do not disappear in crossings. In the infected lines, in seven experiments out of thirty (20% of cases) significant sex-specific influence of Wolbachia on LG was revealed, namely, in two combinations at the optimal temperature 25 degrees C, and in five combinations at stress temperatures. Under the optimal temperature conditions the endosymbiont shortened its host LG in both cases, while at stress temperatures, in four cases out of five, it extended the host LG. Females of the long-living line Bi90 (wMel) from a mountain Kirghiz population appeared to be especially sensitive to the infection carriage--their endosymbiont-caused LG extension was observed both under cold and heat stresses. This result is analogues to the effect of diminishing the oncosupressor lgl dose. In both cases LG extends only under temperature stress, and this positive effect is sex-specific.}, } @article {pmid19851452, year = {2009}, author = {Kremer, N and Voronin, D and Charif, D and Mavingui, P and Mollereau, B and Vavre, F}, title = {Wolbachia interferes with ferritin expression and iron metabolism in insects.}, journal = {PLoS pathogens}, volume = {5}, number = {10}, pages = {e1000630}, pmid = {19851452}, issn = {1553-7374}, mesh = {Aedes/genetics/growth & development/metabolism/microbiology ; Animals ; Base Sequence ; Chromosome Mapping ; Drosophila/genetics/growth & development/metabolism/microbiology ; Female ; Ferritins/*genetics/metabolism ; Gene Expression Regulation ; Genetic Loci ; Host-Parasite Interactions/genetics ; Insecta/*genetics/growth & development/*metabolism/microbiology ; Iron/*metabolism/pharmacology ; Molecular Sequence Data ; Nucleic Acid Conformation ; Rickettsiaceae Infections/genetics/metabolism ; Symbiosis/genetics ; Wasps/genetics/growth & development/metabolism/microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an intracellular bacterium generally described as being a facultative reproductive parasite. However, Wolbachia is necessary for oogenesis completion in the wasp Asobara tabida. This dependence has evolved recently as a result of interference with apoptosis during oogenesis. Through comparative transcriptomics between symbiotic and aposymbiotic individuals, we observed a differential expression of ferritin, which forms a complex involved in iron storage. Iron is an essential element that is in limited supply in the cell. However, it is also a highly toxic precursor of Reactive Oxygen Species (ROS). Ferritin has also been shown to play a key role in host-pathogen interactions. Measuring ferritin by quantitative RT-PCR, we confirmed that ferritin was upregulated in aposymbiotic compared to symbiotic individuals. Manipulating the iron content in the diet, we showed that iron overload markedly affected wasp development and induced apoptotic processes during oogenesis in A. tabida, suggesting that the regulation of iron homeostasis may also be related to the obligate dependence of the wasp. Finally, we demonstrated that iron metabolism is influenced by the presence of Wolbachia not only in the obligate mutualism with A. tabida, but also in facultative parasitism involving Drosophila simulans and in Aedes aegypti cells. In these latter cases, the expression of Wolbachia bacterioferritin was also increased in the presence of iron, showing that Wolbachia responds to the concentration of iron. Our results indicate that Wolbachia may generally interfere with iron metabolism. The high affinity of Wolbachia for iron might be due to physiological requirement of the bacterium, but it could also be what allows the symbiont to persist in the organism by reducing the labile iron concentration, thus protecting the cell from oxidative stress and apoptosis. These findings also reinforce the idea that pathogenic, parasitic and mutualistic intracellular bacteria all use the same molecular mechanisms to survive and replicate within host cells. By impacting the general physiology of the host, the presence of a symbiont may select for host compensatory mechanisms, which extends the possible consequences of persistent endosymbiont on the evolution of their hosts.}, } @article {pmid19825290, year = {2009}, author = {Jia, FX and Yang, MS and Yang, WJ and Wang, JJ}, title = {Influence of continuous high temperature conditions on Wolbachia infection frequency and the fitness of Liposcelis tricolor (Psocoptera: Liposcelididae).}, journal = {Environmental entomology}, volume = {38}, number = {5}, pages = {1365-1372}, doi = {10.1603/022.038.0503}, pmid = {19825290}, issn = {1938-2936}, mesh = {Animals ; *Hot Temperature ; Insect Control/methods ; Insecta/growth & development/*microbiology/physiology ; Longevity ; Reproduction/physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Endosymbiotic bacteria that potentially influence reproduction and other fitness-related traits of their hosts are widespread in arthropods, and their appeal to researchers' interest is growing. In this study, the influence of continuous high temperature conditions on Wolbachia infection frequency and the fitness of Liposcelis tricolor (Psocoptera: Liposcelididae) was studied in a laboratory. The results showed that the Wolbachia infection frequency was gradually decreased when L. tricolor was reared at 33 degrees C; after six generations of treatment, no Wolbachia wsp gene product was detected, suggesting that the Wolbachia infection was completely eliminated. The combined immature development periods and survival rates of L. tricolor did not vary significantly among six generations; however, the female longevities and fecundities dramatically declined from F(1) through F(6), resulting in decreasing values of population intrinsic rate of increase (r(m)). Using r(m) values, the fitness for F(2), F(3), F(4), F(5), and F(6) relative to F(1) was calculated as 0.995, 0.953, 0.811, 0.700, and 0.552, respectively. We realize that it is not possible to distinguish between the effects of high temperature may have on the metabolic processes of the psocids and the effects of temperature-induced reduction in bacterial infection frequencies. Our study confirms that the combined use of antibiotics and heat treatment is a good control measure for psocids.}, } @article {pmid19825106, year = {2009}, author = {Pfarr, KM and Debrah, AY and Specht, S and Hoerauf, A}, title = {Filariasis and lymphoedema.}, journal = {Parasite immunology}, volume = {31}, number = {11}, pages = {664-672}, pmid = {19825106}, issn = {1365-3024}, mesh = {Animals ; Elephantiasis, Filarial/*complications/parasitology ; Humans ; Immune Tolerance ; Immunity, Innate ; Lymphatic Vessels/metabolism/physiopathology ; Lymphedema/*etiology/metabolism/physiopathology ; Vascular Endothelial Growth Factor A/immunology/metabolism ; Wolbachia/immunology ; }, abstract = {Among the causes of lymphoedema (LE), secondary LE due to filariasis is the most prevalent. It affects only a minority of the 120 million people infected with the causative organisms of lymphatic filariasis (LF), Wuchereria bancrofti and Brugia malayi/timori, but is clustered in families, indicating a genetic basis for development of this pathology. The majority of infected individuals develop filarial-specific immunosuppression that starts even before birth in cases where mothers are infected and is characterized by regulatory T-cell responses and high levels of IgG4, thus tolerating high parasite loads and microfilaraemia. In contrast, individuals with this pathology show stronger immune reactions biased towards Th1, Th2 and probably also Th17. Importantly, as for the aberrant lymph vessel development, innate immune responses that are triggered by the filarial antigen ultimately result in the activation of vascular endothelial growth factors (VEGF), thus promoting lymph vessel hyperplasia as a first step to lymphoedema development. Wolbachia endosymbionts are major inducers of these responses in vitro, and their depletion by doxycycline in LF patients reduces plasma VEGF and soluble VEGF-receptor-3 levels to those seen in endemic normals preceding pathology improvement. The search for the immunogenetic basis for LE could lead to the identification of risk factors and thus, to prevention; and has so far led to the identification of single-nucleotide polymorphisms (SNP) with potential functional relevance to VEGF, cytokine and toll-like receptor (TLR) genes. Hydrocele, a pathology with some similarity to LE in which both lymph vessel dilation and lymph extravasation are shared sequelae, has been found to be strongly associated with a VEGF-A SNP known for upregulation of this (lymph-)angiogenesis factor.}, } @article {pmid19820149, year = {2009}, author = {Suh, E and Mercer, DR and Fu, Y and Dobson, SL}, title = {Pathogenicity of life-shortening Wolbachia in Aedes albopictus after transfer from Drosophila melanogaster.}, journal = {Applied and environmental microbiology}, volume = {75}, number = {24}, pages = {7783-7788}, pmid = {19820149}, issn = {1098-5336}, mesh = {Aedes/genetics/*microbiology/*physiology ; Animals ; Cytoplasm/microbiology ; Drosophila melanogaster/*microbiology ; Embryo, Nonmammalian/microbiology ; Female ; Fertility ; Longevity ; Male ; Mosquito Control ; Symbiosis ; Wolbachia/genetics/*pathogenicity/*physiology ; }, abstract = {Maternally inherited Wolbachia bacteria have evolved mechanisms to manipulate the reproduction of their invertebrate hosts, promoting infection spread. A high fitness cost to the host is maladaptive for obligate endosymbionts, and prior studies show rapid selection of new Wolbachia associations toward commensal or mutualistic symbioses. Here, wMelPop Wolbachia is transferred from Drosophila melanogaster into the mosquito Aedes albopictus. Characterization of the resulting strain provides an extreme example of Wolbachia as a pathogen. In addition to reduced longevity and fecundity, abnormally high Wolbachia density is associated with embryonic mortality that masks the typical pattern of cytoplasmic incompatibility. The results are consistent with earlier reports that show unpredictable shifts in the Wolbachia phenotype after interspecific transfer, which can complicate proposed strategies to modify the age structure of medically important vector populations.}, } @article {pmid19815891, year = {2009}, author = {Mand, S and Pfarr, K and Sahoo, PK and Satapathy, AK and Specht, S and Klarmann, U and Debrah, AY and Ravindran, B and Hoerauf, A}, title = {Macrofilaricidal activity and amelioration of lymphatic pathology in bancroftian filariasis after 3 weeks of doxycycline followed by single-dose diethylcarbamazine.}, journal = {The American journal of tropical medicine and hygiene}, volume = {81}, number = {4}, pages = {702-711}, doi = {10.4269/ajtmh.2009.09-0155}, pmid = {19815891}, issn = {1476-1645}, mesh = {Adolescent ; Adult ; Animals ; Anthelmintics/administration & dosage/therapeutic use ; Anti-Bacterial Agents/administration & dosage/therapeutic use ; Diethylcarbamazine/*administration & dosage/*therapeutic use ; Double-Blind Method ; Doxycycline/*administration & dosage/*therapeutic use ; Drug Administration Schedule ; Elephantiasis, Filarial/*drug therapy ; Humans ; Male ; Middle Aged ; Wuchereria bancrofti ; Young Adult ; }, abstract = {In a placebo controlled trial, the effects of 21- and 10-day doxycycline treatments (200 mg/day) followed by single dose diethylcarbamazine (administered 4 months post treatment) on depletion of Wolbachia endobacteria from Wuchereria bancrofti, filaricidal activity, and amerlioration of scrotal lymph vessel dilation were studied in 57 men from Orissa, India. The 21-day doxycycline course reduced Wolbachia in W. bancrofti by 94% before diethylcarbamazine administration. After 12 months, all patients with this treatment were amicrofilaremic and different from the 10-day doxycycline (42.9%) and placebo (37.5%) groups, and significantly fewer were positive for scrotal worm nests (6.7%) compared with 10-day doxycycline (60%) and placebo (66.7%). Average scrotal lymph vessel diameters were reduced from 0.7 cm pre-treatment to 0.02 cm in patients after 21 days of treatment, while no significant changes were seen in the other groups. This latter feature confirms the beneficial effects of doxycycline on lymphatic dilation and thus adds to the existing evidence that doxycycline, in addition to being macrofilaricidal, may be used to prevent or reverse lymphatic pathology.}, } @article {pmid19812617, year = {2010}, author = {Harris, LR and Kelly, SE and Hunter, MS and Perlman, SJ}, title = {Population dynamics and rapid spread of Cardinium, a bacterial endosymbiont causing cytoplasmic incompatibility in Encarsia pergandiella (Hymenoptera: Aphelinidae).}, journal = {Heredity}, volume = {104}, number = {3}, pages = {239-246}, doi = {10.1038/hdy.2009.130}, pmid = {19812617}, issn = {1365-2540}, mesh = {Animals ; Bacteroidetes/genetics/*physiology ; Female ; Infertility ; Male ; Molecular Sequence Data ; Population Dynamics ; *Symbiosis ; Wasps/genetics/*microbiology/*physiology ; }, abstract = {Cytoplasmic incompatibility (CI) is a common phenotype of maternally inherited bacterial symbionts of arthropods; in its simplest expression, uninfected females produce few or no viable progeny when mated to infected males. Infected females thus experience a reproductive advantage relative to that of uninfected females, with the potential for the symbiont to spread rapidly. CI population dynamics are predicted to depend primarily on the strength of incompatibility, the fitness cost of the infection and how faithfully symbionts are inherited. Although the bacterial symbiont lineage Wolbachia has been most identified with the CI phenotype, an unrelated bacterium, Cardinium may also cause CI. In the first examination of population dynamics of CI-inducing Cardinium, we used population cages of the parasitic wasp Encarsia pergandiella (Hymenoptera: Aphelinidae) with varying initial infection frequencies to test a model of invasion. Cardinium was found to spread rapidly in all populations, even in cases where the initial infection frequency was well below the predicted invasion threshold frequency. The discrepancy between the modeled and actual results is best explained by weaker CI than measured in the lab and a cryptic fitness benefit to the infection.}, } @article {pmid19812409, year = {2009}, author = {Hoerauf, A}, title = {Mansonella perstans--the importance of an endosymbiont.}, journal = {The New England journal of medicine}, volume = {361}, number = {15}, pages = {1502-1504}, doi = {10.1056/NEJMe0905193}, pmid = {19812409}, issn = {1533-4406}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; Elephantiasis, Filarial/complications/drug therapy ; Filaricides/*therapeutic use ; Humans ; Mansonella ; Mansonelliasis/complications/*drug therapy ; Rickettsiaceae Infections/*complications/drug therapy ; *Symbiosis ; *Wolbachia ; Wuchereria bancrofti/isolation & purification ; }, } @article {pmid19812401, year = {2009}, author = {Coulibaly, YI and Dembele, B and Diallo, AA and Lipner, EM and Doumbia, SS and Coulibaly, SY and Konate, S and Diallo, DA and Yalcouye, D and Kubofcik, J and Doumbo, OK and Traore, AK and Keita, AD and Fay, MP and Traore, SF and Nutman, TB and Klion, AD}, title = {A randomized trial of doxycycline for Mansonella perstans infection.}, journal = {The New England journal of medicine}, volume = {361}, number = {15}, pages = {1448-1458}, pmid = {19812401}, issn = {1533-4406}, support = {ZIA AI001063-04/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Adolescent ; Adult ; Aged ; Albendazole/therapeutic use ; Animals ; Anti-Bacterial Agents/administration & dosage/adverse effects/*therapeutic use ; Doxycycline/administration & dosage/adverse effects/*therapeutic use ; Drug Therapy, Combination ; Elephantiasis, Filarial/complications/drug therapy ; Female ; Filaricides/*therapeutic use ; Humans ; Ivermectin/therapeutic use ; Male ; *Mansonella/isolation & purification ; Mansonelliasis/complications/*drug therapy ; Middle Aged ; Rickettsiaceae Infections/complications/*drug therapy ; Symbiosis ; Treatment Outcome ; *Wolbachia ; Wuchereria bancrofti/isolation & purification ; Young Adult ; }, abstract = {BACKGROUND: Mansonella perstans infection is common in areas of Africa where Wuchereria bancrofti, a causative agent of lymphatic filariasis, is endemic. M. perstans is refractory to standard antifilarial therapies. The recent discovery of bacterial endosymbionts (e.g., wolbachia) in most filarial species, including M. perstans, provides new therapeutic options for reducing microfilaremia.

METHODS: In an open-label, randomized trial, we recruited subjects with M. perstans microfilaremia, with or without concomitant W. bancrofti infection, from four villages in Mali and randomly assigned them to receive doxycycline, at a dose of 200 mg daily for 6 weeks (106 subjects), or no treatment (110). At 6 months, subjects who were coinfected with W. bancrofti underwent a second random assignment, to treatment with a single dose of albendazole (400 mg) and ivermectin (150 microg per kilogram of body weight) or no treatment. Subjects were monitored daily during the first 6-week study period for adverse events. M. perstans and W. bancrofti microfilarial levels were assessed at 6, 12, and 36 months.

RESULTS: At 12 months, 67 of 69 subjects who had received treatment with doxycycline only (97%) had no detectable M. perstans microfilariae per 60 microl of blood, as compared with 10 of 63 subjects who had received no treatment (16%) (relative risk, 6.18; 95% confidence interval, 3.63 to 11.89; P<0.001). At 36 months, M. perstans microfilaremia remained suppressed in 48 of 64 subjects who had received treatment with doxycycline only (75%), a finding that was consistent with a macrofilaricidal effect of doxycycline. Vomiting was more frequent in the doxycycline-treated group than in the untreated group (17% vs. 4%).

CONCLUSIONS: These results are consistent with previous findings that M. perstans harbors the intracellular endosymbiont, wolbachia, and suggest that doxycycline is an effective therapy for M. perstans infection. (ClinicalTrials.gov number, NCT00340691.)}, } @article {pmid19806204, year = {2009}, author = {Ghedin, E and Hailemariam, T and DePasse, JV and Zhang, X and Oksov, Y and Unnasch, TR and Lustigman, S}, title = {Brugia malayi gene expression in response to the targeting of the Wolbachia endosymbiont by tetracycline treatment.}, journal = {PLoS neglected tropical diseases}, volume = {3}, number = {10}, pages = {e525}, pmid = {19806204}, issn = {1935-2735}, support = {R01 AI072465/AI/NIAID NIH HHS/United States ; AI072465/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/*drug effects/*genetics/microbiology/physiology ; Female ; Gene Expression Regulation/*drug effects ; Helminth Proteins/genetics ; *Symbiosis ; Tetracycline/*pharmacology ; Wolbachia/*drug effects/physiology ; }, abstract = {BACKGROUND: Brugia malayi, like most human filarial parasite species, harbors an endosymbiotic bacterium of the genus Wolbachia. Elimination of the endosymbiont leads to sterilization of the adult female. Previous biochemical and genetic studies have established that communication with its endobacterium is essential for survival of the worm.

We used electron microscopy to examine the effects of antibiotic treatment on Wolbachia cell structure. We have also used microarray and quantitative RT-PCR analyses to examine the regulation of the B. malayi transcripts altered in response to the anti-Wolbachia treatment. Microscopy of worms taken from animals treated with tetracycline for 14 and 21 days (14 d and 21 d) demonstrated substantial morphologic effects on the Wolbachia endobacterium by 14 d and complete degeneration of the endobacterial structures by 21 d. We observed upregulation of transcripts primarily encoding proteins involved in amino acid synthesis and protein translation, and downregulation of transcripts involved in cuticle biosynthesis after both 7 d and 14 d of treatment. In worms exposed to tetracycline in culture, substantial effects on endobacteria morphology were evident by day 3, and extensive death of the endobacteria was observed by day 5. In a detailed examination of the expression kinetics of selected signaling genes carried out on such cultured worms, a bimodal pattern of regulation was observed. The selected genes were upregulated during the early phase of antibiotic treatment and quickly downregulated in the following days. These same genes were upregulated once more at 6 days post-treatment.

CONCLUSIONS/SIGNIFICANCE: Upregulation of protein translation and amino acid synthesis may indicate a generalized stress response induced in B. malayi due to a shortage of essential nutrients/factors that are otherwise supplied by Wolbachia. Downregulation of transcripts involved in cuticle biosynthesis perhaps reflects a disruption in the normal embryogenic program. This is confirmed by the expression pattern of transcripts that may be representative of the worms' response to Wolbachia in different tissues; the early peak potentially reflects the effect of bacteria death on the embryogenic program while the second peak may be a manifestation of the adult worm response to the affected bacteria within the hypodermis.}, } @article {pmid19800031, year = {2009}, author = {Geiger, A and Fardeau, ML and Grebaut, P and Vatunga, G and Josénando, T and Herder, S and Cuny, G and Truc, P and Ollivier, B}, title = {First isolation of Enterobacter, Enterococcus, and Acinetobacter spp. as inhabitants of the tsetse fly (Glossina palpalis palpalis) midgut.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {9}, number = {6}, pages = {1364-1370}, doi = {10.1016/j.meegid.2009.09.013}, pmid = {19800031}, issn = {1567-7257}, mesh = {Acinetobacter/cytology/*isolation & purification/physiology ; Angola ; Animals ; DNA, Bacterial/genetics/isolation & purification ; Enterobacter/cytology/*isolation & purification/physiology ; Enterococcus/cytology/*isolation & purification/physiology ; Gastrointestinal Tract/*microbiology ; Humans ; Insect Vectors/microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/analysis/genetics ; Symbiosis ; Trypanosomiasis, African/transmission ; Tsetse Flies/*microbiology/physiology ; }, abstract = {This paper reports the first evidence of the presence of bacteria, other than the three previously described as symbionts, Wigglesworthia glossinidia, Wolbachia, and Sodalis glossinidius, in the midgut of Glossina palpalis palpalis, the tsetse fly, a vector of the chronic form of human African trypanosomiasis in sub-Saharan African countries. Based on the morphological, nutritional, physiological, and phylogenetic results, we identified Enterobacter, Enterococcus, and Acinetobacter spp. as inhabitants of the midgut of the tsetse fly from Angola. Enterobacter spp. was the most frequently isolated. The role of these bacteria in the gut, in terms of vector competence of the tsetse fly, is discussed, as is the possibility of using these bacteria to produce in situ trypanolytic molecules.}, } @article {pmid19797660, year = {2009}, author = {Kambris, Z and Cook, PE and Phuc, HK and Sinkins, SP}, title = {Immune activation by life-shortening Wolbachia and reduced filarial competence in mosquitoes.}, journal = {Science (New York, N.Y.)}, volume = {326}, number = {5949}, pages = {134-136}, pmid = {19797660}, issn = {1095-9203}, support = {079059/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/genetics/immunology/*microbiology/*parasitology ; Animals ; Brugia pahangi/growth & development/*physiology ; Elephantiasis, Filarial/prevention & control/transmission ; Genes, Insect ; Host-Parasite Interactions ; Immunity, Innate/*genetics ; Insect Vectors/immunology/microbiology/parasitology ; Longevity ; Mosquito Control ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {Wolbachia strain wMelPop reduces the longevity of its Drosophila melanogaster host and, when introduced into the mosquito Aedes aegypti, halves its life span. We show that wMelPop induces up-regulation of the mosquito's innate immune system and that its presence inhibits the development of filarial nematodes in the mosquito. These data suggest that wMelPop could be used in the global effort to eliminate lymphatic filariasis and possibly for the control of other mosquito-borne parasites where immune preactivation inhibits their development. The cost of constitutive immune up-regulation may contribute to the life-shortening phenotype.}, } @article {pmid19773076, year = {2009}, author = {Vavre, F and Mouton, L and Pannebakker, BA}, title = {Drosophila-parasitoid communities as model systems for host-Wolbachia interactions.}, journal = {Advances in parasitology}, volume = {70}, number = {}, pages = {299-331}, doi = {10.1016/S0065-308X(09)70012-0}, pmid = {19773076}, issn = {2163-6079}, mesh = {Animals ; Biological Evolution ; Drosophila/*parasitology ; Drosophila melanogaster/parasitology ; Female ; Genetic Variation ; Host-Parasite Interactions/*physiology ; Male ; *Models, Biological ; Oogenesis/physiology ; Phenotype ; Phylogeny ; Wasps/microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are cytoplasmic endosymbionts that infect a wide range of arthropod and nematode hosts. They are transmitted from mother to offspring via the eggs (vertical transmission) and enhance their transmission to the next generation by manipulating the reproductive system of their hosts. These manipulations occur in many forms, such as the induction of cytoplasmic incompatibility, feminization, male killing and parthenogenesis induction. Wolbachia is estimated to occur in up to 66% of all insect species, but the greatest diversity of reproductive manipulations is found in the order of the Hymenoptera. Studies of Wolbachia in Drosophila-parasitoid communities have allowed for important insights into different aspects of Wolbachia biology. The extensive knowledge available on Drosophila parasitoids provides a solid base on which to test new hypotheses on host-Wolbachia interactions. The large range of Wolbachia phenotypes present in Drosophila parasitoids, combined with the recent acquisition of the bacteria from their Drosophilid hosts, make them an ideal model system to study the evolution and dynamics of Wolbachia infections, both in the laboratory as in the field. In this chapter, we aim to review the current knowledge on the associations between Wolbachia and Drosophila parasitoids, and identify open questions and specify new research directions.}, } @article {pmid19758348, year = {2009}, author = {Bressan, A and Arneodo, J and Simonato, M and Haines, WP and Boudon-Padieu, E}, title = {Characterization and evolution of two bacteriome-inhabiting symbionts in cixiid planthoppers (Hemiptera: Fulgoromorpha: Pentastirini).}, journal = {Environmental microbiology}, volume = {11}, number = {12}, pages = {3265-3279}, doi = {10.1111/j.1462-2920.2009.02055.x}, pmid = {19758348}, issn = {1462-2920}, mesh = {Animals ; Biological Evolution ; Female ; Gammaproteobacteria/genetics/*isolation & purification/ultrastructure ; Hemiptera/genetics/metabolism/*microbiology ; *Metagenome ; Phylogeny ; RNA, Ribosomal, 16S/metabolism ; *Symbiosis ; }, abstract = {Like other plant sap-sucking insects, planthoppers within the family Cixiidae (Insecta: Hemiptera: Fulgoromorpha) host a diversified microbiota. We report the identification and first molecular characterization of symbiotic bacteria in cixiid planthoppers (tribe: Pentastirini). Using universal eubacterial primers we first screened the eubacterial 16S rRNA sequences in Pentastiridius leporinus (Linnaeus) with PCR amplification, cloning, and restriction fragment analysis. We identified three main 16S rRNA sequences that corresponded to a Wolbachia bacterium, a plant pathogenic bacterium, and a novel gammaproteobacterial symbiont. A fourth bacterial species affiliated with 'Candidatus Sulcia muelleri' was detected in PCR assays using primers specific for the Bacteroidetes. Within females of two selected cixiid planthoppers, P. leporinus and Oliarus filicicola, fluorescence In situ hybridization analysis and transmission electron microscopy observations showed that 'Ca. Sulcia muelleri' and the novel gammaproteobacterial symbiont were housed in separate bacteriomes. Phylogenetic analysis revealed that both of these symbionts occurred in at least four insect genera within the tribe Pentastirini. 'Candidatus Purcelliella pentastirinorum' was proposed as the novel gammaproteobacterial symbiont.}, } @article {pmid19756742, year = {2009}, author = {Specht, S and Hoerauf, A and Adjei, O and Debrah, A and Büttner, DW}, title = {Newly acquired Onchocerca volvulus filariae after doxycycline treatment.}, journal = {Parasitology research}, volume = {106}, number = {1}, pages = {23-31}, pmid = {19756742}, issn = {1432-1955}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Female ; Humans ; Male ; Onchocerca volvulus/anatomy & histology/*isolation & purification ; Onchocerciasis/*drug therapy/*parasitology ; Placebos/administration & dosage ; Wolbachia/drug effects ; }, abstract = {Despite successful mass drug administration and vector control programs, the nematode Onchocerca volvulus is far from being eradicated. Therefore, new long-term sterilizing or macrofilaricidal drugs are needed. The depletion of Wolbachia endobacteria using doxycycline leads to long-term sterilizing effects and macrofilaricidal activity against female filariae of more than 60%. The worms die or degenerate 18-27 months after doxycycline. However, during this time patients may be exposed to new infections. We evaluated these newly acquired worms in onchocercomas of doxycycline-treated patients in relation to transmission using morphology, histochemistry, and immunohistology. On an average, 10% of the female filariae had been newly acquired per year in the treated groups. Our observations showed: (a) Sixty-three of 68 newly acquired worms harbored many intact Wolbachia, whereas none of the other surviving worms contained many bacteria. (b) Higher percentages of dead filariae than originally reported were calculated, when the new worms were excluded, indicating a stronger macrofilaricidal activity than previously reported. The difference was significant for female filariae after doxycycline treatment for 6 weeks. (c) Only newly acquired worms presented normal embryogenesis and microfilariae production after sufficient treatment. We conclude that newly acquired filariae have to be considered when evaluating drug efficacy in onchocerciasis.}, } @article {pmid19753103, year = {2009}, author = {Turley, AP and Moreira, LA and O'Neill, SL and McGraw, EA}, title = {Wolbachia infection reduces blood-feeding success in the dengue fever mosquito, Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {3}, number = {9}, pages = {e516}, pmid = {19753103}, issn = {1935-2735}, abstract = {BACKGROUND: The mosquito Aedes aegypti was recently transinfected with a life-shortening strain of the endosymbiont Wolbachia pipientis (wMelPop) as the first step in developing a biocontrol strategy for dengue virus transmission. In addition to life-shortening, the wMelPop-infected mosquitoes also exhibit increased daytime activity and metabolic rates. Here we sought to quantify the blood-feeding behaviour of Wolbachia-infected females as an indicator of any virulence or energetic drain associated with Wolbachia infection.

In a series of blood-feeding trials in response to humans, we have shown that Wolbachia-infected mosquitoes do not differ in their response time to humans, but that as they age they obtain fewer and smaller blood meals than Wolbachia-uninfected controls. Lastly, we observed a behavioural characteristic in the Wolbachia infected mosquitoes best described as a "bendy" proboscis that may explain the decreased biting success.

CONCLUSIONS/SIGNIFICANCE: Taken together the evidence suggests that wMelPop infection may be causing tissue damage in a manner that intensifies with mosquito age and that leads to reduced blood-feeding success. These behavioural changes require further investigation with respect to a possible physiological mechanism and their role in vectorial capacity of the insect. The selective decrease of feeding success in older mosquitoes may act synergistically with other Wolbachia-associated traits including life-shortening and viral protection in biocontrol strategies.}, } @article {pmid19747825, year = {2009}, author = {Hornett, EA and Charlat, S and Wedell, N and Jiggins, CD and Hurst, GD}, title = {Rapidly shifting sex ratio across a species range.}, journal = {Current biology : CB}, volume = {19}, number = {19}, pages = {1628-1631}, doi = {10.1016/j.cub.2009.07.071}, pmid = {19747825}, issn = {1879-0445}, mesh = {Animals ; Butterflies/genetics/*parasitology ; DNA Primers/genetics ; Female ; Malaysia ; Male ; Philippines ; Polynesia ; Sequence Analysis, DNA ; *Sex Ratio ; Time Factors ; *Wolbachia ; }, abstract = {Sex ratios are subject to distortion by a range of inherited parasites. Although it has been predicted that the presence of these elements will result in spatial and temporal variation in host sex ratio, testing of this hypothesis has been constrained by availability of historical data. We here determine spatial and temporal variation in sex ratio in a interaction between a butterfly and male-killing Wolbachia bacteria by assaying infection presence in museum specimens, and from this inferring infection prevalence and phenotype in historical populations. Comparison of contemporary and museum samples revealed profound change in four of five populations examined. Two populations become extremely female biased, associated with spread of the male-killer bacterium. One evolved from extremely female biased to a sex ratio near parity, resulting from the infection losing male-killing activity. The final population fluctuated widely in sex ratio, associated with varying frequency of the male killer. We conclude that asynchronous invasion and decline of sex-ratio distorters combines with the evolution of host suppressors to produce a rapidly changing mosaic of sex ratio. As a consequence, the reproductive ecology of the host species is likely to be fundamentally altered over short time scales. Further, the study demonstrates the utility of museum specimens as "silent witnesses" of evolutionary change.}, } @article {pmid19738635, year = {2010}, author = {Zabal-Aguirre, M and Arroyo, F and Bella, JL}, title = {Distribution of Wolbachia infection in Chorthippus parallelus populations within and beyond a Pyrenean hybrid zone.}, journal = {Heredity}, volume = {104}, number = {2}, pages = {174-184}, doi = {10.1038/hdy.2009.106}, pmid = {19738635}, issn = {1365-2540}, mesh = {Animals ; Female ; Grasshoppers/classification/*microbiology/*physiology ; Male ; Molecular Sequence Data ; Phylogeny ; Sexual Behavior, Animal ; Spain ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Two subspecies of the meadow grasshopper Chorthippus parallelus meet in a secondary hybrid zone in the Pyrenees. We have recently detected Wolbachia infection in this extensively studied species. C. p. parallelus (Cpp) and C. p. erythropus (Cpe) harbour bacteria from the B and F supergroups, but they differ noticeably in the incidence and type of infection present in their populations. We can distinguish different regional infection patterns that are associated with the distribution of pure and hybrid C. parallelus individuals. The northern pattern of low-level infection characterizes Cpp populations, whereas the southern pattern of high-level infection affects Cpe locations. These patterns converge in the hybrid zone and generate a third infection pattern featuring an extremely high degree of co-infection with the two Wolbachia types in hybrids. The distribution of Wolbachia among the studied populations encourages us to consider the possibility that this bacterium has a significant influence on the origin, maintenance and dynamics of this hybrid zone, given the reproductive alterations that are often associated with Wolbachia.}, } @article {pmid19734338, year = {2009}, author = {Nakamura, Y and Kawai, S and Yukuhiro, F and Ito, S and Gotoh, T and Kisimoto, R and Yanase, T and Matsumoto, Y and Kageyama, D and Noda, H}, title = {Prevalence of Cardinium bacteria in planthoppers and spider mites and taxonomic revision of "Candidatus Cardinium hertigii" based on detection of a new Cardinium group from biting midges.}, journal = {Applied and environmental microbiology}, volume = {75}, number = {21}, pages = {6757-6763}, pmid = {19734338}, issn = {1098-5336}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Infections/microbiology/*veterinary ; Bacteroidetes/*classification/*isolation & purification ; Cluster Analysis ; DNA Gyrase/genetics ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {Cardinium bacteria, members of the phylum Cytophaga-Flavobacterium-Bacteroides (CFB), are intracellular bacteria in arthropods that are capable of inducing reproductive abnormalities in their hosts, which include parasitic wasps, mites, and spiders. A high frequency of Cardinium infection was detected in planthoppers (27 out of 57 species were infected). A high frequency of Cardinium infection was also found in spider mites (9 out of 22 species were infected). Frequencies of double infection by Cardinium and Wolbachia bacteria (Alphaproteobacteria capable of manipulating reproduction of their hosts) were disproportionately high in planthoppers but not in spider mites. A new group of bacteria, phylogenetically closely related to but distinct from previously described Cardinium bacteria (based on 16S rRNA and gyrB genes) was found in 4 out of 25 species of Culicoides biting midges. These bacteria possessed a microfilament-like structure that is a morphological feature previously found in Cardinium and Paenicardinium. The bacteria close to the genus Cardinium consist of at least three groups, A, B, and C. Group A is present in various species of arthropods and was previously referred to as "Candidatus Cardinium hertigii," group B is present in plant parasitic nematodes and was previously referred to as "Candidatus Paenicardinium endonii," and group C is present in Culicoides biting midges. On the basis of morphological and molecular data, we propose that the nomenclature of these three groups be integrated into a single species, "Candidatus Cardinium hertigii."}, } @article {pmid19732336, year = {2009}, author = {Arthofer, W and Riegler, M and Schneider, D and Krammer, M and Miller, WJ and Stauffer, C}, title = {Hidden Wolbachia diversity in field populations of the European cherry fruit fly, Rhagoletis cerasi (Diptera, Tephritidae).}, journal = {Molecular ecology}, volume = {18}, number = {18}, pages = {3816-3830}, doi = {10.1111/j.1365-294X.2009.04321.x}, pmid = {19732336}, issn = {1365-294X}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Europe ; Female ; *Genetic Variation ; *Genetics, Population ; Geography ; Male ; Plasmids/genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Tephritidae/*microbiology ; Wolbachia/classification/*genetics ; }, abstract = {The European cherry fruit fly Rhagoletis cerasi has been a field model for cytoplasmic incompatibility since the mid 1970s. Two Wolbachia strains were detected in this tephritid species and wCer2 was described as the CI inducing agent dividing European populations into two unidirectional incompatible groups, i.e. southern females produce viable offspring with northern males, whereas the reciprocal cross results in incompatibility. We detected three new Wolbachia strains by sequencing a multitude of plasmids derived from Wolbachia surface protein gene (wsp) polymerase chain reaction (PCR) products. Strain-specific primers were developed allowing individual diagnosis without need for cloning. Hybridization of specific PCR products with a wsp oligonucleotide enhanced the detection limit significantly and revealed the presence of low-titre infections in some strains, in different ontogenetic stages and in adults of different age. We then performed a survey of strain prevalence and infection frequency in eight European regions. wCer1 was fixed in all populations, whereas wCer2 was detected only in the South. wCer3 frequency was the lowest without a clear distribution pattern. The abundance of wCer4 was homogenous across Europe. Like wCer2, wCer5 showed significant differences in spatial distribution. Our new findings of previously undetected and recombinant Wolbachia strains in R. cerasi reveal a major caveat to the research community not to overlook hidden Wolbachia diversity in field populations. Low-titres and geographical variability in Wolbachia diversity are expected to influence the outcome of Wolbachia population dynamics and Wolbachia-based insect population control and may create invasion barriers for expanding and artificially introduced Wolbachia strains.}, } @article {pmid19731208, year = {2009}, author = {Gazla, IN and Carracedo, MC}, title = {Effect of intracellular Wolbachia on interspecific crosses between Drosophila melanogaster and Drosophila simulans.}, journal = {Genetics and molecular research : GMR}, volume = {8}, number = {3}, pages = {861-869}, doi = {10.4238/vol8-3gmr595}, pmid = {19731208}, issn = {1676-5680}, mesh = {Analysis of Variance ; Animals ; *Crosses, Genetic ; Drosophila/*genetics/*microbiology ; Drosophila melanogaster/*genetics/*microbiology ; Female ; Hybridization, Genetic ; Intracellular Space/*microbiology ; Male ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia are bacteria that live inside the cells of a large number of invertebrate hosts and are transmitted from infected females to their offspring. Their presence is associated with cytoplasmic incompatibility in several species of Drosophila. Cytoplasmic incompatibility results when the sperm of infected males fertilize eggs of uninfected females, causing more or less intense embryonic mortality (unidirectional incompatibility). This phenomenon also appears in crosses between populations infected with different Wolbachia strains (bidirectional incompatibility). The influence of Wolbachia infection on host populations has attracted attention as a potentially rapid mechanism for development of reproductive isolation and subsequent speciation. We examined the influence of this bacterium on reproductive isolation in interspecific crosses between Drosophila melanogaster and D. simulans. We found that Wolbachia infection negatively affected these two species in homospecific crosses. However, in interspecific crosses, it only influenced sexual isolation, as infected females more frequently hybridized than females free of infection; postzygotic reduction of fitness (bidirectional cytoplasmic incompatibility) was not detected. This would be explained by the existence of several modes of rescue systems in these two species, reducing cytoplasmic incompatibility between them. Wolbachia does not appear to cause reproductive isolation between these two species.}, } @article {pmid19722191, year = {2009}, author = {Dangi, A and Vedi, S and Nag, JK and Paithankar, S and Singh, MP and Kar, SK and Dube, A and Misra-Bhattacharya, S}, title = {Tetracycline treatment targeting Wolbachia affects expression of an array of proteins in Brugia malayi parasite.}, journal = {Proteomics}, volume = {9}, number = {17}, pages = {4192-4208}, doi = {10.1002/pmic.200800324}, pmid = {19722191}, issn = {1615-9861}, mesh = {Animals ; Antibodies, Helminth ; Brugia malayi/drug effects/immunology/*metabolism/*microbiology ; Electrophoresis, Gel, Two-Dimensional ; Electrophoresis, Polyacrylamide Gel ; Helminth Proteins/*metabolism ; Immunoblotting ; Parasites/drug effects/*metabolism/*microbiology ; Polymerase Chain Reaction ; Proteome/analysis ; Reproducibility of Results ; Tetracycline/*pharmacology ; Wolbachia/*drug effects ; }, abstract = {Wolbachia is an intracellular endosymbiont of Brugia malayi parasite whose presence is essential for the survival of the parasite. Treatment of B. malayi-infected jirds with tetracycline eliminates Wolbachia, which affects parasite survival and fitness. In the present study we have tried to identify parasite proteins that are affected when Wolbachia is targeted by tetracycline. For this Wolbachia depleted parasites (B. malayi) were obtained by tetracycline treatment of infected Mongolian jirds (Meriones unguiculatus) and their protein profile after 2-DE separation was compared with that of untreated parasites harboring Wolbachia. Approximately 100 protein spots could be visualized followed by CBB staining of 2-D gel and included for comparative analysis. Of these, 54 showed differential expressions, while two new protein spots emerged (of 90.3 and 64.4 kDa). These proteins were subjected to further analysis by MALDI-TOF for their identification using Brugia coding sequence database composed of both genomic and EST sequences. Our study unravels two crucial findings: (i) the parasite or Wolbachia proteins, which disappeared/down-regulated appear be essential for parasite survival and may be used as drug targets and (ii) tetracycline treatment interferes with the regulatory machinery vital for parasites cellular integrity and defense and thus could possibly be a molecular mechanism for the killing of filarial parasite. This is the first proteomic study substantiating the wolbachial genome integrity with its nematode host and providing functional genomic data of human lymphatic filarial parasite B. malayi.}, } @article {pmid19720566, year = {2009}, author = {Simón, F and Morchón, R and González-Miguel, J and Marcos-Atxutegi, C and Siles-Lucas, M}, title = {What is new about animal and human dirofilariosis?.}, journal = {Trends in parasitology}, volume = {25}, number = {9}, pages = {404-409}, doi = {10.1016/j.pt.2009.06.003}, pmid = {19720566}, issn = {1471-5007}, mesh = {Animals ; *Cat Diseases/epidemiology/parasitology/physiopathology/transmission ; Cats/*parasitology ; Climate Change ; *Dirofilaria/classification/microbiology/pathogenicity/physiology ; Dirofilaria immitis/microbiology/pathogenicity/physiology ; *Dirofilariasis/epidemiology/parasitology/physiopathology/transmission ; *Dog Diseases/epidemiology/parasitology/physiopathology/transmission ; Dogs/*parasitology ; *Host-Parasite Interactions/immunology ; Humans ; Wolbachia/immunology ; }, abstract = {Dirofilaria immitis and D. repens, the causal agents of cardiopulmonary and subcutaneous dirofilariosis, respectively, affect canine, feline and human populations with an increasing incidence in temperate and tropical areas of the world. In the past decade outstanding advances in the knowledge of dirofilariosis have been achieved. Nevertheless, questions such as the impact of climate change in the transmission and distribution of dirofilariosis, as well as a profound evaluation of both the role of Dirofilaria and Wolbachia and the proteins produced by them in the parasite-host relationship have not been fully addressed; therefore there must be milestones in dirofilariosis research in order to design new strategies and tools for the control of this disease.}, } @article {pmid19710065, year = {2009}, author = {Koop, JL and Zeh, DW and Bonilla, MM and Zeh, JA}, title = {Reproductive compensation favours male-killing Wolbachia in a live-bearing host.}, journal = {Proceedings. Biological sciences}, volume = {276}, number = {1675}, pages = {4021-4028}, pmid = {19710065}, issn = {0962-8452}, mesh = {Animals ; Arachnida/genetics/*microbiology/*physiology ; Female ; Hydrogen-Ion Concentration ; Male ; Phylogeny ; Reproduction/physiology ; Sex Ratio ; Wolbachia/*classification ; }, abstract = {Wolbachia are maternally inherited, cellular endosymbionts that can enhance their fitness by biasing host sex ratio in favour of females. Male killing (MK) is an extreme form of sex-ratio manipulation that is selectively advantageous if the self-sacrifice of Wolbachia in males increases transmission through females. In live-bearing hosts, females typically produce more embryos than can be carried to term, and reproductive compensation through maternal resource reallocation from dead males to female embryos could increase the number of daughters born to infected females. Here, we report a new strain of MK Wolbachia (wCsc2) in the pseudoscorpion, Cordylochernes scorpioides, and present the first empirical evidence that reproductive compensation favours the killing of males in a viviparous host. Females infected with the wCsc2 strain produced 26 per cent more and significantly larger daughters than tetracycline-cured females. In contrast to the previously described wCsc1 MK Wolbachia strain in C. scorpioides, wCsc2 infection was not accompanied by an increase in the rate of spontaneous brood abortion. Characterization of the wCsc1 and wCsc2 strains by multi-locus sequence typing and by Wolbachia surface protein (wsp) gene sequencing indicates that the marked divergence between these two MK strains in their impact on host reproductive success, and hence in their potential to spread, has occurred in association with homologous recombination in the wsp gene.}, } @article {pmid19703224, year = {2010}, author = {Debout, GD and Frederickson, ME and Aron, S and Yu, DW}, title = {Unexplained split sex ratios in the neotropical plant-ant, Allomerus octoarticulatus var. demerarae (Myrmicinae): a test of hypotheses.}, journal = {Evolution; international journal of organic evolution}, volume = {64}, number = {1}, pages = {126-141}, doi = {10.1111/j.1558-5646.2009.00824.x}, pmid = {19703224}, issn = {1558-5646}, mesh = {Animals ; Ants/genetics/*physiology ; Female ; Male ; Microsatellite Repeats/genetics ; *Sex Ratio ; }, abstract = {We investigated sex allocation in the Neotropical ant Allomerus octoarticulatus var. demerarae. Because Allomerus is a plant symbiont, we could make geographically extensive collections of complete colonies and of foundresses in saplings, allowing us to estimate not only population- and colony-level sex allocation but also colony resource levels and the relatedness of competing ant foundresses. This species exhibits a strongly split sex ratio, with 80% of mature colonies producing >or=90% of one sex or the other. Our genetic analyses (DNA microsatellites) reveal that Allomerus has a breeding system characterized by almost complete monogyny and a low frequency of polyandry. Contrary to theoretical explanations, we find no difference in worker relatedness asymmetries between female- and male-specialist colonies. Furthermore, no clear link was found between colony sex allocation and life history traits such as the number of mates per queen, or colony size, resource level, or fecundity. We also failed to find significant support for male production by workers, infection by Wolbachia, local resource competition, or local mate competition. We are left with the possibility that Allomerus exhibits split sex ratios because of the evolution of alternative biasing strategies in queens or workers, as recently proposed in the literature.}, } @article {pmid19698196, year = {2010}, author = {Kawasaki, Y and Ito, M and Miura, K and Kajimura, H}, title = {Superinfection of five Wolbachia in the alnus ambrosia beetle, Xylosandrus germanus (Blandford) (Coleoptera: Curuculionidae).}, journal = {Bulletin of entomological research}, volume = {100}, number = {2}, pages = {231-239}, doi = {10.1017/S000748530999023X}, pmid = {19698196}, issn = {1475-2670}, mesh = {Animals ; Coleoptera/*microbiology ; Electron Transport Complex IV/genetics ; Genes, Bacterial/genetics ; Haplotypes ; Phylogeny ; Polymorphism, Genetic ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are among the most common endosymbionts in insects. In Wolbachia research, the Wolbachia surface protein (wsp) gene has been used as a phylogenetic tool, but relationships inferred by single-locus analysis can be unreliable because of the extensive genome recombination among Wolbachia strains. Therefore, a multilocus sequence typing (MLST) method for Wolbachia, which relies upon a set of five conserved genes, is recommended. In this study, we examined whether the alnus ambrosia beetle, Xylosandrus germanus (Blandford), is infected with Wolbachia using wsp and MLST genes. Wolbachia was detected from all tested specimens of X. germanus (n=120) by wsp amplification. Five distinct sequences (i.e. five alleles) for wsp were found, and labeled as wXge1-5. MLST analysis and molecular phylogeny of concatenated sequences of MLST genes identified wXge3 and wXge5 as closely-related strains. The detection rate of wXge4 and wXge1 was 100% and 63.3%, respectively; wXge2, wXge3 and wXge5 were detected from less than 15% of specimens. We performed mitochondrial haplotype analyses that identified three genetic types of X. germanus, i.e. Clades A, B and C. Wsp alleles wXge1, wXge2 and wXge4 were detected in all clade A beetles; wXge2 allele was absent from Clades B and C. We concluded that (i) five wsp alleles were found from X. germanus, (ii) use of MLST genes, rather than the wsp gene, are more suited to construct Wolbachia phylogenies and (iii) wsp alleles wXge2 and wXge3/wXge5 would infect clade A and clade B/C of X. germanus, respectively.}, } @article {pmid19698188, year = {2009}, author = {Pike, N and Kingcombe, R}, title = {Antibiotic treatment leads to the elimination of Wolbachia endosymbionts and sterility in the diplodiploid collembolan Folsomia candida.}, journal = {BMC biology}, volume = {7}, number = {}, pages = {54}, pmid = {19698188}, issn = {1741-7007}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Arthropods/chemistry/*microbiology/physiology ; Cloning, Organism ; Clutch Size/drug effects ; Denmark ; Diet ; Diploidy ; Female ; France ; Male ; Ovum/drug effects ; Parthenogenesis ; Polymerase Chain Reaction ; Reproduction/drug effects ; Rifampin/*pharmacology ; Symbiosis/*drug effects ; Tetracycline/*pharmacology ; United Kingdom ; Wolbachia/chemistry/*drug effects/physiology ; }, abstract = {BACKGROUND: Wolbachia is an extremely widespread bacterial endosymbiont of arthropods and nematodes that causes a variety of reproductive peculiarities. Parthenogenesis is one such peculiarity but it has been hypothesised that this phenomenon may be functionally restricted to organisms that employ haplodiploid sex determination. Using two antibiotics, tetracycline and rifampicin, we attempted to eliminate Wolbachia from the diplodiploid host Folsomia candida, a species of springtail which is a widely used study organism.

RESULTS: Molecular assays confirmed that elimination of Wolbachia was successfully achieved through continuous exposure of populations (over two generations and several weeks) to rifampicin administered as 2.7% dry weight of their yeast food source. The consequence of this elimination was total sterility of all individuals, despite the continuation of normal egg production.

CONCLUSION: Microbial endosymbionts play an obligatory role in the reproduction of their diplodiploid host, most likely one in which the parthenogenetic process is facilitated by Wolbachia. A hitherto unknown level of host-parasite interdependence is thus recorded.}, } @article {pmid19692410, year = {2009}, author = {Ikeya, T and Broughton, S and Alic, N and Grandison, R and Partridge, L}, title = {The endosymbiont Wolbachia increases insulin/IGF-like signalling in Drosophila.}, journal = {Proceedings. Biological sciences}, volume = {276}, number = {1674}, pages = {3799-3807}, pmid = {19692410}, issn = {0962-8452}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila melanogaster/*metabolism/*microbiology ; Fat Body/metabolism ; Gene Expression Regulation ; Insulin/*metabolism ; Mutation ; Signal Transduction ; Somatomedins/*metabolism ; Wolbachia/*physiology ; }, abstract = {Insulin/IGF-like signalling (IIS) is an evolutionarily conserved pathway that has diverse functions in multi-cellular organisms. Mutations that reduce IIS can have pleiotropic effects on growth, development, metabolic homeostasis, fecundity, stress resistance and lifespan. IIS is also modified by extrinsic factors. For instance, in the fruitfly Drosophila melanogaster, both nutrition and stress can alter the activity of the pathway. Here, we test experimentally the hypothesis that a widespread endosymbiont of arthropods, Wolbachia pipientis, can alter the degree to which mutations in genes encoding IIS components affect IIS and its resultant phenotypes. Wolbachia infection, which is widespread in D. melanogaster in nature and has been estimated to infect 30 per cent of strains in the Bloomington stock centre, can affect broad aspects of insect physiology, particularly traits associated with reproduction. We measured a range of IIS-related phenotypes in flies ubiquitously mutant for IIS in the presence and absence of Wolbachia. We show that removal of Wolbachia further reduces IIS and hence enhances the mutant phenotypes, suggesting that Wolbachia normally acts to increase insulin signalling. This effect of Wolbachia infection on IIS could have an evolutionary explanation, and has some implications for studies of IIS in Drosophila and other organisms that harbour endosymbionts.}, } @article {pmid19692404, year = {2009}, author = {Aikawa, T and Anbutsu, H and Nikoh, N and Kikuchi, T and Shibata, F and Fukatsu, T}, title = {Longicorn beetle that vectors pinewood nematode carries many Wolbachia genes on an autosome.}, journal = {Proceedings. Biological sciences}, volume = {276}, number = {1674}, pages = {3791-3798}, pmid = {19692404}, issn = {0962-8452}, mesh = {Animals ; Coleoptera/*microbiology/*parasitology ; Female ; Genes, Bacterial ; Genome, Insect ; Male ; Nematoda/*physiology ; Phylogeny ; Wolbachia/*genetics ; }, abstract = {Monochamus alternatus is the longicorn beetle notorious as a vector of the pinewood nematode that causes the pine wilt disease. When two populations of M. alternatus were subjected to diagnostic polymerase chain reaction (PCR) detection of four Wolbachia genes, only the ftsZ gene was detected from one of the populations. The Wolbachia ftsZ gene persisted even after larvae were fed with a tetracycline-containing diet for six weeks. The inheritance of the ftsZ gene was not maternal but biparental, exhibiting a typical Mendelian pattern. The ftsZ gene titres in homozygotic ftsZ(+) insects were nearly twice as high as those in heterozygotic ftsZ(+) insects. Exhaustive PCR surveys revealed that 31 and 30 of 214 Wolbachia genes examined were detected from the two insect populations, respectively. Many of these Wolbachia genes contained stop codon(s) and/or frame shift(s). Fluorescent in situ hybridization confirmed the location of the Wolbachia genes on an autosome. On the basis of these results, we conclude that a large Wolbachia genomic region has been transferred to and located on an autosome of M. alternatus. The discovery of massive gene transfer from Wolbachia to M. alternatus would provide further insights into the evolution and fate of laterally transferred endosymbiont genes in multicellular host organisms.}, } @article {pmid19686755, year = {2009}, author = {Wiwatanaratanabutr, I and Kittayapong, P}, title = {Effects of crowding and temperature on Wolbachia infection density among life cycle stages of Aedes albopictus.}, journal = {Journal of invertebrate pathology}, volume = {102}, number = {3}, pages = {220-224}, doi = {10.1016/j.jip.2009.08.009}, pmid = {19686755}, issn = {1096-0805}, mesh = {Aedes/anatomy & histology/growth & development/*microbiology ; Animals ; Female ; Larva/growth & development/microbiology ; Male ; Population Density ; Temperature ; Wolbachia/*physiology ; }, abstract = {Species of the genus Wolbachia are a group of Rickettsia-like, maternally-inherited bacteria (gram negative), which cause various reproductive alterations in their arthropod and nematode hosts including cytoplasmic incompatibility (CI), male-killing, parthenogenesis and feminization. They can be divided into supergroups such as A and B based on phylogenetic analysis of 16S rDNA sequences. In this study, we examined the relative infection densities of Wolbachia strains among life cycle stages in the mosquito, Aedes albopictus in terms of crowding effect and temperature effect. A. albopictus is known to be superinfected with both A- and B-supergroup Wolbachia which cause CI. The relative Wolbachia densities within each individual mosquito were determined and quantified by using real-time quantitative PCR assay based on the wsp gene. We found that B-supergroup Wolbachia strain densities in this host species were consistently and significantly higher than in the A-supergroup. Larval crowding also reduced adult size of mosquitoes. Our results show clearly that the higher densities of mosquito larvae cause lower densities of Wolbachia strains. Examination of the effect of temperature on Wolbachia density in each stage of the mosquito clearly revealed a significant decrease in bacterial density following exposure to elevated temperature (37 degrees C) in both males and females.}, } @article {pmid19686264, year = {2010}, author = {Turelli, M}, title = {Cytoplasmic incompatibility in populations with overlapping generations.}, journal = {Evolution; international journal of organic evolution}, volume = {64}, number = {1}, pages = {232-241}, doi = {10.1111/j.1558-5646.2009.00822.x}, pmid = {19686264}, issn = {1558-5646}, mesh = {Aging ; Animals ; Cytoplasm/immunology/*physiology ; Female ; Insecta/microbiology/*physiology ; Male ; Wolbachia/isolation & purification ; }, abstract = {Many insects and other arthropods harbor maternally inherited bacteria inducing "cytoplasmic incompatibility" (CI), reduced egg hatch when infected males mate with uninfected females. CI-causing infections produce a frequency-dependent reproductive advantage for infected females. However, many such infections impose fitness costs that lead to unstable equilibrium frequencies below which the infections tend to be eliminated. To understand the unstable equilibria produced by reduced lifespan or lengthened development, overlapping-generation analyses are needed. An idealized model of overlapping generations with age-independent parameters produces a simple expression showing how the unstable point depends on the population growth rate, the intensity of CI, and the infection's effects on development time, longevity, and fecundity. The interpretation of this equilibrium is complicated by age structure. Nevertheless, the unstable equilibrium provides insight into the CI-causing infections found in nature, and it can guide potential manipulations of natural populations, including those that transmit diseases, through the introduction of infections that alter life-table parameters.}, } @article {pmid19682363, year = {2009}, author = {Brelsfoard, CL and St Clair, W and Dobson, SL}, title = {Integration of irradiation with cytoplasmic incompatibility to facilitate a lymphatic filariasis vector elimination approach.}, journal = {Parasites & vectors}, volume = {2}, number = {1}, pages = {38}, pmid = {19682363}, issn = {1756-3305}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; R01 AI067434/AI/NIAID NIH HHS/United States ; }, abstract = {BACKGROUND: Mass drug administration (MDA) is the emphasis of an ongoing global lymphatic filariasis (LF) elimination program by the World Health Organization, in which the entire 'at risk' human population is treated annually with anti-filarial drugs. However, there is evidence that the MDA strategy may not be equally appropriate in all areas of LF transmission, leading to calls for the augmentation of MDA with anti-vector interventions. One potential augmentative intervention is the elimination of vectors via repeated inundative releases of male mosquitoes made cytoplasmically incompatible via an infection with Wolbachia bacteria. However, with a reduction in the vector population size, there is the risk that an accidental female release would permit the establishment of the incompatible Wolbachia infection type, resulting in population replacement instead of population elimination. To avoid the release of fertile females, we propose the exposure of release individuals to low doses of radiation to sterilize any accidentally released females, reducing the risk of population replacement.

RESULTS: Aedes polynesiensis pupae of differing ages were irradiated to determine a radiation dose that results in sterility but that does not affect the survival and competitiveness of males. Laboratory assays demonstrate that males irradiated at a female sterilizing dosage of 40 Gy are equally competitive with un-irradiated males. No effect of irradiation on the ability of Wolbachia to affect egg hatch was observed.

CONCLUSION: An irradiation dose of 40 Gy is sufficient to cause female sterility, but has no observed negative effect on male fitness. The results support further development of this approach as a preventative measure against accidental population replacement.}, } @article {pmid19663639, year = {2009}, author = {Matsuura, Y and Koga, R and Nikoh, N and Meng, XY and Hanada, S and Fukatsu, T}, title = {Huge symbiotic organs in giant scale insects of the genus Drosicha (Coccoidea: Monophlebidae) harbor flavobacterial and enterobacterial endosymbionts.}, journal = {Zoological science}, volume = {26}, number = {7}, pages = {448-456}, doi = {10.2108/zsj.26.448}, pmid = {19663639}, issn = {0289-0003}, mesh = {Animals ; Enterobacteriaceae/genetics/*isolation & purification/ultrastructure ; Flavobacteriaceae/genetics/*isolation & purification/ultrastructure ; Hemiptera/*anatomy & histology/*microbiology ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Symbiosis/*physiology ; }, abstract = {Giant scale insects (Drosicha: Coccoldea: Monophlebidae) were investigated for their symbiotic organs and bacterial endosymbionts. Two types of bacterial 16S rRNA gene sequences, flavobacterial and enterobacterial, were consistently detected in D. corpulenta and D. pinicola. The former sequences formed a compact clade in the Bacteroidetes, allied to the symbionts of cushion and armored scales. The latter sequences formed a robust clade in the gamma-Proteobacteria, allied to enteric bacteria like Enterobacter aerogenes and Escherichia coli. Another type of 16S sequence derived from Wolbachia was also detected in D. pinicola. In-situ hybridization demonstrated that the flavobacterial and enterobacterial symbionts were localized in a pair of huge bacteriomes in the abdomen, the former in uninucleated peripheral bacteriocytes and the latter in syncytial central bacteriocytes. Electron microscopy confirmed the endocellular locations of the pleomorphic flavobacterial symbiont and the rod-shaped enterobacterial symbiont, and also revealed the location and fine structure of the Wolbachia symbiont in D. pinicola. Infection frequencies of the flavobacterial and enterobacterial symbionts were consistently 100% in populations of D. corpulenta and D. pinicola, while the Wolbachia symbiont exhibited 0% and 100% infection frequencies in D. corpulente and D. pinicola, respectively. Neither the flavobacterial symbiont nor the enterobacterial symbiont exhibited AT-biased nucleotide composition or accelerated molecular evolution. The huge bacteriomes of Drosicha giant scales would provide a useful system for investigating biochemical, physiological, and genomic aspects of the host-symbiont and symbiont-symbiont interactions.}, } @article {pmid19659591, year = {2009}, author = {Kraaijeveld, K and Franco, P and Reumer, BM and van Alphen, JJ}, title = {Effects of parthenogenesis and geographic isolation on female sexual traits in a parasitoid wasp.}, journal = {Evolution; international journal of organic evolution}, volume = {63}, number = {12}, pages = {3085-3096}, doi = {10.1111/j.1558-5646.2009.00798.x}, pmid = {19659591}, issn = {1558-5646}, mesh = {Animals ; Chromatography, Gas ; Female ; Geography ; Male ; *Parthenogenesis ; Sexual Behavior, Animal ; Spermatozoa ; Wasps/*physiology ; }, abstract = {Population divergence in sexual traits is affected by different selection pressures, depending on the mode of reproduction. In allopatric sexual populations, aspects of sexual behavior may diverge due to sexual selection. In parthenogenetic populations, loss-of-function mutations in genes involved in sexual functionality may be selectively neutral or favored by selection. We assess to what extent these processes have contributed to divergence in female sexual traits in the parasitoid wasp Leptopilina clavipes in which some populations are infected with parthenogenesis-inducing Wolbachia bacteria. We find evidence consistent with both hypotheses. Both arrhenotokous males and males derived from thelytokous strains preferred to court females from their own population. This suggests that these populations had already evolved population-specific mating preferences when the latter became parthenogenetic. Thelytokous females did not store sperm efficiently and fertilized very few of their eggs. The nonfertility of thelytokous females was due to mutations in the wasp genome, which must be an effect of mutation accumulation under thelytoky. Divergence in female sexual traits of these two allopatric populations has thus been molded by different forces: independent male/female coevolution while both populations were still sexual, followed by female-only evolution after one population switched to parthenogenesis.}, } @article {pmid19656295, year = {2009}, author = {Henrichfreise, B and Schiefer, A and Schneider, T and Nzukou, E and Poellinger, C and Hoffmann, TJ and Johnston, KL and Moelleken, K and Wiedemann, I and Pfarr, K and Hoerauf, A and Sahl, HG}, title = {Functional conservation of the lipid II biosynthesis pathway in the cell wall-less bacteria Chlamydia and Wolbachia: why is lipid II needed?.}, journal = {Molecular microbiology}, volume = {73}, number = {5}, pages = {913-923}, doi = {10.1111/j.1365-2958.2009.06815.x}, pmid = {19656295}, issn = {1365-2958}, mesh = {Alkyl and Aryl Transferases/antagonists & inhibitors ; Bacterial Outer Membrane Proteins/isolation & purification/metabolism ; Bacterial Proteins/isolation & purification/metabolism ; Biosynthetic Pathways/*genetics ; Cell Membrane/metabolism ; Chlamydia/drug effects/*genetics/*metabolism ; Enzyme Inhibitors/pharmacology ; Fosfomycin/pharmacology ; Genes, Bacterial ; Genes, Essential ; Models, Biological ; Monosaccharides/metabolism ; N-Acetylglucosaminyltransferases/isolation & purification/metabolism ; Oligopeptides/metabolism ; Transferases/isolation & purification/metabolism ; Transferases (Other Substituted Phosphate Groups) ; Uridine Diphosphate N-Acetylmuramic Acid/*analogs & derivatives/biosynthesis ; Wolbachia/drug effects/*genetics/*metabolism ; }, abstract = {Cell division and cell wall biosynthesis in prokaryotes are driven by partially overlapping multiprotein machineries whose activities are tightly controlled and co-ordinated. So far, a number of protein components have been identified and acknowledged as essential for both fundamental cellular processes. Genes for enzymes of both machineries have been found in the genomes of the cell wall-less genera Chlamydia and Wolbachia, raising questions as to the functionality of the lipid II biosynthesis pathway and reasons for its conservation. We provide evidence on three levels that the lipid II biosynthesis pathway is indeed functional and essential in both genera: (i) fosfomycin, an inhibitor of MurA, catalysing the initial reaction in lipid II biosynthesis, has a detrimental effect on growth of Wolbachia cells; (ii) isolated cytoplasmic membranes from Wolbachia synthesize lipid II ex vivo; and (iii) recombinant MraY and MurG from Chlamydia and Wolbachia exhibit in vitro activity, synthesizing lipid I and lipid II respectively. We discuss the hypothesis that the necessity for maintaining lipid II biosynthesis in cell wall-lacking bacteria reflects an essential role of the precursor in prokaryotic cell division. Our results also indicate that the lipid II pathway may be exploited as an antibacterial target for chlamydial and filarial infections.}, } @article {pmid19654607, year = {2010}, author = {Echaubard, P and Duron, O and Agnew, P and Sidobre, C and Noël, V and Weill, M and Michalakis, Y}, title = {Rapid evolution of Wolbachia density in insecticide resistant Culex pipiens.}, journal = {Heredity}, volume = {104}, number = {1}, pages = {15-19}, doi = {10.1038/hdy.2009.100}, pmid = {19654607}, issn = {1365-2540}, mesh = {Animals ; Culex/genetics/*microbiology ; Female ; Genome, Bacterial/genetics ; Genome, Insect/genetics ; Genotype ; Host-Pathogen Interactions ; Insecticide Resistance/genetics/*physiology ; Male ; Polymerase Chain Reaction ; Population Density ; Sex Factors ; Wolbachia/genetics/growth & development/*physiology ; }, abstract = {The maternally inherited symbiotic Wolbachia have been previously shown to have much greater densities in insecticide-resistant Culex pipiens mosquitoes than in insecticide-susceptible individuals. These high densities were shown to be at least partially responsible for the costs related to insecticide resistance in this species. We report here the rapid evolution, on the order of 50 generations, of bacterial densities both in laboratory and field populations. Along with other recently published studies, this report shows that Wolbachia-host interactions are very dynamic.}, } @article {pmid19653892, year = {2009}, author = {Branca, A and Vavre, F and Silvain, JF and Dupas, S}, title = {Maintenance of adaptive differentiation by Wolbachia induced bidirectional cytoplasmic incompatibility: the importance of sib-mating and genetic systems.}, journal = {BMC evolutionary biology}, volume = {9}, number = {}, pages = {185}, pmid = {19653892}, issn = {1471-2148}, mesh = {Adaptation, Biological/genetics ; Alleles ; Animals ; Computer Simulation ; Diploidy ; Evolution, Molecular ; Female ; Genome, Insect ; Haploidy ; Male ; *Models, Genetic ; Reproduction/genetics ; *Selection, Genetic ; Wasps/*genetics/microbiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Bacteria of the genus Wolbachia are reproductive parasites widespread among arthropods. The most common effect arising from the presence of Wolbachia in a population is Cytoplasmic Incompatibility (CI), whereby postmating reproductive isolation occurs in crosses between an infected male and an uninfected female, or when a male is infected with a different strain of Wolbachia to that of the female (bidirectional CI). Previous theoretical models have demonstrated that bidirectional CI can contribute to the genetic divergence of populations in haploid and diploid organisms. However, haplodiploid organisms were not considered in these models even though they include Nasonia parasitoid wasps - the best example of the implication of Wolbachia in ongoing speciation. Moreover, previous work did not investigate inbreeding mating systems, which are frequently observed in arthropod species.

RESULTS: We developed a stochastic two-island model which simulated three genetic scenarios, diploidy, haploidy, and haplodiploidy, with two CI phenotypes being considered for the latter: (1) male development of female progeny; and (2) mortality of fertilized eggs. We also investigated the effect of varying the proportion of sib mating. In the model each allopatric population was initially fixed for a single allele at a nuclear locus under positive selection and infected with one strain of Wolbachia. Each simulation presupposed that the two populations were fixed for a different allele and a different strain of Wolbachia. The degree of genetic differentiation observed in the locus under selection due to bidirectional CI was much lower for the two haplodiploid phenotypes than for either diploids or haploids. Furthermore, we demonstrated that sib-mating may compensate for the lower efficiency of bidirectional CI in haplodiploids by maintaining genetic divergence.

CONCLUSION: Our model suggests that maintenance of genetic differentiation facilitated by Wolbachia is more likely to occur in diploids and haploids than in haplodiploids. However, increasing the level of sib-mating may compensate for the weak effect of bidirectional CI in haplodiploids. Our work therefore gives a potential explanation for why the haplodiploid Nasonia species, which are infected with bidirectionally incompatible Wolbachia strains and undergo sib-mating, have differentiated genetically and maintained this differentiation without premating isolation.}, } @article {pmid19652990, year = {2009}, author = {Borgo, SN and Sattler, EC and Hogardt, M and Adler, K and Plewig, G}, title = {PCR analysis for Wolbachia in human and canine Demodex mites.}, journal = {Archives of dermatological research}, volume = {301}, number = {10}, pages = {747-752}, doi = {10.1007/s00403-009-0984-0}, pmid = {19652990}, issn = {1432-069X}, mesh = {Animals ; Anti-Inflammatory Agents/therapeutic use ; DNA, Bacterial/*analysis ; Dogs ; Folliculitis ; Gram-Negative Bacterial Infections/diagnosis/drug therapy/*genetics/pathology/physiopathology ; Humans ; Mite Infestations/diagnosis/drug therapy/*microbiology/pathology/physiopathology ; Mites/*microbiology/pathogenicity ; Polymerase Chain Reaction ; Rosacea ; Skin/*metabolism/microbiology/pathology ; Symbiosis ; Tetracyclines/therapeutic use ; Wolbachia/*genetics/pathogenicity ; }, abstract = {In many skin diseases such as Demodex folliculitis, rosacea- or steroid-induced rosacea Demodex mites are present in abundance and are at least partially held responsible for causing these disorders. Although it is known that these diseases respond well to tetracyclines, it is unclear if this is due to the antiinflammatory effects of the antibiotics or to an antibacterial effect on so far unknown bacteria within the Demodex mites. As in filariasis, where the response to doxycycline can be explained by the presence of Wolbachia within the filarial nematodes, this study was performed to see whether Wolbachia also use Demodex mites as their hosts. Human and canine Demodex mite samples were taken by skin scrapings and tested by PCR for the presence of Wolbachia DNA. Wolbachia pipientis DNA was used as positive control. In none of the DNA extracts, Wolbachia were detected showing no evidence for the presence of these bacteria in Demodex mites. The response of Demodex aggravated or Demodex caused diseases to tetracyclines seems not to be due to the presence of Wolbachia in Demodex mites in contrast to the results seen in filariasis.}, } @article {pmid19640319, year = {2009}, author = {Smith, DT and Hosken, DJ and Ffrench-Constant, RH and Wedell, N}, title = {Variation in sex peptide expression in D. melanogaster.}, journal = {Genetics research}, volume = {91}, number = {4}, pages = {237-242}, doi = {10.1017/S0016672309000226}, pmid = {19640319}, issn = {1469-5073}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Drosophila Proteins/*genetics ; Drosophila melanogaster/*genetics/microbiology/physiology ; Female ; *Gene Expression Profiling ; *Genetic Variation ; Host-Pathogen Interactions ; Male ; Peptides/*genetics ; Regression Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Sexual Behavior, Animal ; Species Specificity ; Wolbachia/physiology ; }, abstract = {Male Drosophila melanogaster transfers many accessory-gland proteins to females during copulation. Sex peptide (SP) is one of these and one of its main effects is to decrease female remating propensity. To date, there has been no investigation of genetic variation in SP-gene expression levels, or if such potential variation directly influences female remating behaviour. We assessed both these possibilities and found significant variation in expression levels of the SP gene across D. melanogaster isolines. A non-linear association between SP expression levels and female remating delay suggestive of disruptive selection on expression levels was also documented. Finally, while some isolines were infected with the endosymbiont Wolbachia, no association between Wolbachia and SP expression level was found.}, } @article {pmid19633721, year = {2009}, author = {Zouache, K and Voronin, D and Tran-Van, V and Mousson, L and Failloux, AB and Mavingui, P}, title = {Persistent Wolbachia and cultivable bacteria infection in the reproductive and somatic tissues of the mosquito vector Aedes albopictus.}, journal = {PloS one}, volume = {4}, number = {7}, pages = {e6388}, pmid = {19633721}, issn = {1932-6203}, mesh = {Aedes/*microbiology ; Animals ; Base Sequence ; DNA Primers ; Female ; Genitalia, Female/*microbiology ; In Situ Hybridization, Fluorescence ; *Insect Vectors ; Polymerase Chain Reaction ; *Reproduction ; Wolbachia/*isolation & purification ; }, abstract = {BACKGROUND: Commensal and symbiotic microbes have a considerable impact on the behavior of many arthropod hosts, including hematophagous species that transmit pathogens causing infectious diseases to human and animals. Little is known about the bacteria associated with mosquitoes other than the vectorized pathogens. This study investigated Wolbachia and cultivable bacteria that persist through generations in Ae. albopictus organs known to host transmitted arboviruses, such as dengue and chikungunya.

We used culturing, diagnostic and quantitative PCR, as well as in situ hybridization, to detect and locate bacteria in whole individual mosquitoes and in dissected tissues. Wolbachia, cultivable bacteria of the genera Acinetobacter, Comamonas, Delftia and Pseudomonas co-occurred and persisted in the bodies of both males and females of Ae. albopictus initially collected in La Réunion during the chikungunya outbreak, and maintained as colonies in insectaries. In dissected tissues, Wolbachia and the cultivable Acinetobacter can be detected in the salivary glands. The other bacteria are commonly found in the gut. Quantitative PCR estimates suggest that Wolbachia densities are highest in ovaries, lower than those of Acinetobacter in the gut, and approximately equal to those of Acinetobacter in the salivary glands. Hybridization using specific fluorescent probes successfully localized Wolbachia in all germ cells, including the oocytes, and in the salivary glands, whereas the Acinetobacter hybridizing signal was mostly located in the foregut and in the anterior midgut.

CONCLUSIONS/SIGNIFICANCE: Our results show that Proteobacteria are distributed in the somatic and reproductive tissues of mosquito where transmissible pathogens reside and replicate. This location may portend the coexistence of symbionts and pathogens, and thus the possibility that competition or cooperation phenomena may occur in the mosquito vector Ae. albopictus. Improved understanding of the vectorial system, including the role of bacteria in the vector's biology and competence, could have major implications for understanding viral emergences and for disease control.}, } @article {pmid19629359, year = {2009}, author = {Dreyer, G and Mattos, D and Figueredo-Silva, J and Norões, J}, title = {[Paradigm shift in bancroftian filariasis].}, journal = {Revista da Associacao Medica Brasileira (1992)}, volume = {55}, number = {3}, pages = {355-362}, doi = {10.1590/s0104-42302009000300032}, pmid = {19629359}, issn = {0104-4230}, mesh = {Animals ; Diagnosis, Differential ; Elephantiasis, Filarial/*diagnosis/microbiology/parasitology ; Humans ; Wolbachia/physiology ; Wuchereria bancrofti ; }, abstract = {The way a particular subject is understood changes over time as a result of scientific research. In most cases, these changes are minor, with limited effect on the overall knowledge on the subject. Sometimes, however, revolutionary changes occur and not only modify the understanding of the subject but open perspectives that can trigger new interpretations and new ways for expansion of scientific knowledge. The studies of Gregor Johann Mendel were a good example. They led to discovery of the laws of inheritance which, in turn, have revolutionized biology and provided the foundation for genetics. In certain situations, changes not only alter ways of thinking, but have practical implications, also improving the quality of life for many people. In his book The Structure of Scientific Revolutions, Thomas Kuhn refers to discontinuities in scientific development as a 'change of paradigm', a term now used in a generic manner to describe a profound changes in our reference points. For lymphatic filariasis the old paradigm stated that Wuchereria bancrofti at the adult stage causes lymphatic vessel obstruction, triggering an inevitable immune response in predisposed individuals and leading to elephantiasis. This has been replaced by a new paradigm, which offers hope that W. bancrofti infection does not necessarily predispose to the disfiguring outward manifestation of lymphatic dysfunction. Repeated secondary bacterial infections (erysipela-like) are now recognized as the most important factor for initiation and progression of chronic lymphedema in individuals living in filariasis-endemic areas. Most inhabitants of endemic communities can prevent and minimize the acute bacterial episodes by regular use of soap and water, the simplest form of hygiene already well known to human beings.}, } @article {pmid19603576, year = {2009}, author = {Weiner, MP and Slatko, B}, title = {Discover the microbes within! The HHMI Wolbachia Project.}, journal = {BioTechniques}, volume = {46}, number = {7}, pages = {497}, doi = {10.2144/000113182}, pmid = {19603576}, issn = {1940-9818}, mesh = {*Academies and Institutes ; Animals ; Biology/*education ; Education ; Insecta/genetics/*microbiology ; Phylogeny ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*physiology ; }, } @article {pmid19597542, year = {2009}, author = {Wu, B and Novelli, J and Foster, J and Vaisvila, R and Conway, L and Ingram, J and Ganatra, M and Rao, AU and Hamza, I and Slatko, B}, title = {The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target.}, journal = {PLoS neglected tropical diseases}, volume = {3}, number = {7}, pages = {e475}, pmid = {19597542}, issn = {1935-2735}, support = {R01 DK074797/DK/NIDDK NIH HHS/United States ; R01 DK074797-01/DK/NIDDK NIH HHS/United States ; DK074797/DK/NIDDK NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/antagonists & inhibitors/genetics ; Biosynthetic Pathways/*drug effects/genetics ; Brugia malayi/*growth & development/*microbiology/physiology ; Cloning, Molecular ; Cluster Analysis ; Drug Evaluation, Preclinical/methods ; Enzyme Inhibitors/pharmacology ; Escherichia coli/drug effects/genetics ; Female ; Genetic Complementation Test ; Heme/*biosynthesis/genetics ; Humans ; Locomotion ; Male ; Phylogeny ; Sequence Homology ; Wolbachia/*drug effects/isolation & purification/*metabolism ; }, abstract = {BACKGROUND: Filarial parasites (e.g., Brugia malayi, Onchocerca volvulus, and Wuchereria bancrofti) are causative agents of lymphatic filariasis and onchocerciasis, which are among the most disabling of neglected tropical diseases. There is an urgent need to develop macro-filaricidal drugs, as current anti-filarial chemotherapy (e.g., diethylcarbamazine [DEC], ivermectin and albendazole) can interrupt transmission predominantly by killing microfilariae (mf) larvae, but is less effective on adult worms, which can live for decades in the human host. All medically relevant human filarial parasites appear to contain an obligate endosymbiotic bacterium, Wolbachia. This alpha-proteobacterial mutualist has been recognized as a potential target for filarial nematode life cycle intervention, as antibiotic treatments of filarial worms harboring Wolbachia result in the loss of worm fertility and viability upon antibiotic treatments both in vitro and in vivo. Human trials have confirmed this approach, although the length of treatments, high doses required and medical counter-indications for young children and pregnant women warrant the identification of additional anti-Wolbachia drugs.

METHODS AND FINDINGS: Genome sequence analysis indicated that enzymes involved in heme biosynthesis might constitute a potential anti-Wolbachia target set. We tested different heme biosynthetic pathway inhibitors in ex vivo B. malayi viability assays and report a specific effect of N-methyl mesoporphyrin (NMMP), which targets ferrochelatase (FC, the last step). Our phylogenetic analysis indicates evolutionarily significant divergence between Wolbachia heme genes and their human homologues. We therefore undertook the cloning, overexpression and analysis of several enzymes of this pathway alongside their human homologues, and prepared proteins for drug targeting. In vitro enzyme assays revealed a approximately 600-fold difference in drug sensitivities to succinyl acetone (SA) between Wolbachia and human 5'-aminolevulinic acid dehydratase (ALAD, the second step). Similarly, Escherichia coli hemH (FC) deficient strains transformed with human and Wolbachia FC homologues showed significantly different sensitivities to NMMP. This approach enables functional complementation in E. coli heme deficient mutants as an alternative E. coli-based method for drug screening.

CONCLUSIONS: Our studies indicate that the heme biosynthetic genes in the Wolbachia of B. malayi (wBm) might be essential for the filarial host survival. In addition, the results suggest they are likely candidate drug targets based upon significant differences in phylogenetic distance, biochemical properties and sensitivities to heme biosynthesis inhibitors, as compared to their human homologues.}, } @article {pmid19595661, year = {2009}, author = {Hasan, AU and Suguri, S and Ahmed, SM and Fujimoto, C and Harada, M and Rahman, SM and Zaman, RU and Kakehi, Y}, title = {Molecular phylogeography of Culex quinquefasciatus mosquitoes in central Bangladesh.}, journal = {Acta tropica}, volume = {112}, number = {2}, pages = {106-114}, doi = {10.1016/j.actatropica.2009.07.005}, pmid = {19595661}, issn = {1873-6254}, mesh = {Acetylcholinesterase/genetics ; Animals ; Bangladesh ; Cluster Analysis ; Culex/*classification/*genetics ; DNA, Ribosomal Spacer ; Electron Transport Complex IV/genetics ; Evolution, Molecular ; Molecular Sequence Data ; Peptide Elongation Factor 1/genetics ; Sequence Analysis, DNA ; Sequence Homology ; Wolbachia/genetics ; }, abstract = {Mosquitoes in the Culex pipiens complex are a major vector of numerous parasitic and arboviral diseases. Here we report the phylogeography of a prevalent Culex mosquito, Cx. quinquefasciatus, from three locations in Bangladesh: Dhaka, Savar and Mymensingh. Sequence analysis of the genes encoding mitochondrial cytochrome oxidase subunit II, nuclear elongation factor-1 alpha, and acetylcholinesterase-2 revealed the lack of a population genetic structure among the three locations. Moreover, the highly divergent ribosomal internal transcribed spacer 2 suggests that this locus has not evolved in concert. The results further show evidence of historical introgression of internal transcribed spacer 2 from Cx. pipiens to Cx. quinquefasciatus of Bangladesh, and that the introgression occurred before Cx. quinquefasciatus had dispersed within this region. The study also reveals historical population expansion in this region, followed by a post-expansion Wolbachia sweep.}, } @article {pmid19592535, year = {2009}, author = {Tanaka, K and Furukawa, S and Nikoh, N and Sasaki, T and Fukatsu, T}, title = {Complete WO phage sequences reveal their dynamic evolutionary trajectories and putative functional elements required for integration into the Wolbachia genome.}, journal = {Applied and environmental microbiology}, volume = {75}, number = {17}, pages = {5676-5686}, pmid = {19592535}, issn = {1098-5336}, mesh = {Bacteriophages/*genetics/physiology ; DNA, Viral/*chemistry/genetics ; Evolution, Molecular ; Gene Order ; *Genome, Viral ; Molecular Sequence Data ; Phylogeny ; Prophages/*genetics/physiology ; *Sequence Analysis, DNA ; Sequence Homology ; Viral Proteins/genetics/physiology ; Virus Integration ; Wolbachia/*virology ; }, abstract = {Wolbachia endosymbionts are ubiquitously found in diverse insects including many medical and hygienic pests, causing a variety of reproductive phenotypes, such as cytoplasmic incompatibility, and thereby efficiently spreading in host insect populations. Recently, Wolbachia-mediated approaches to pest control and management have been proposed, but the application of these approaches has been hindered by the lack of genetic transformation techniques for symbiotic bacteria. Here, we report the genome and structure of active bacteriophages from a Wolbachia endosymbiont. From the Wolbachia strain wCauB infecting the moth Ephestia kuehniella two closely related WO prophages, WOcauB2 of 43,016 bp with 47 open reading frames (ORFs) and WOcauB3 of 45,078 bp with 46 ORFs, were characterized. In each of the prophage genomes, an integrase gene and an attachment site core sequence were identified, which are putatively involved in integration and excision of the mobile genetic elements. The 3' region of the prophages encoded genes with sequence motifs related to bacterial virulence and protein-protein interactions, which might represent effector molecules that affect cellular processes and functions of their host bacterium and/or insect. Database searches and phylogenetic analyses revealed that the prophage genes have experienced dynamic evolutionary trajectories. Genes similar to the prophage genes were found across divergent bacterial phyla, highlighting the active and mobile nature of the genetic elements. We suggest that the active WO prophage genomes and their constituent sequence elements would provide a clue to development of a genetic transformation vector for Wolbachia endosymbionts.}, } @article {pmid19545353, year = {2009}, author = {Goodacre, SL and Martin, OY and Bonte, D and Hutchings, L and Woolley, C and Ibrahim, K and George Thomas, C and Hewitt, GM}, title = {Microbial modification of host long-distance dispersal capacity.}, journal = {BMC biology}, volume = {7}, number = {}, pages = {32}, pmid = {19545353}, issn = {1741-7007}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Behavior, Animal ; Female ; Male ; Maternal Exposure ; Population Dynamics ; Rickettsia/drug effects/growth & development ; Spiders/drug effects/*microbiology ; Spiroplasma/drug effects/growth & development ; Symbiosis ; Wolbachia/drug effects/growth & development ; }, abstract = {BACKGROUND: Dispersal plays a key role in shaping biological and ecological processes such as the distribution of spatially-structured populations or the pace and scale of invasion. Here we have studied the relationship between long-distance dispersal behaviour of a pest-controlling money spider, Erigone atra, and the distribution of maternally acquired endosymbionts within the wider meta-population. This spider persists in heterogeneous environments because of its ability to recolonise areas through active long-distance airborne dispersal using silk as a sail, in a process termed 'ballooning'.

RESULTS: We show that there is spatial heterogeneity in the prevalence of two maternally acquired endosymbiont infections within the wider E. atra meta-population and we demonstrate through several independent approaches a link between the presence of one of these endosymbionts, Rickettsia, and the tendency for long-distance movement.

CONCLUSION: This novel finding that particular endosymbionts can influence host dispersal is of broad importance given the extremely widespread occurrence of similar bacteria within arthropod communities. A bacterial phenotype that limits dispersal has the potential not only to reduce gene flow and thus contribute to degrees of reproductive isolation within species, but also to influence species distribution and thus overall community composition.}, } @article {pmid19539785, year = {2009}, author = {Morales-Hojas, R}, title = {Molecular systematics of filarial parasites, with an emphasis on groups of medical and veterinary importance, and its relevance for epidemiology.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {9}, number = {5}, pages = {748-759}, doi = {10.1016/j.meegid.2009.06.007}, pmid = {19539785}, issn = {1567-7257}, mesh = {Animals ; Evolution, Molecular ; Filariasis/*epidemiology/*parasitology ; Filarioidea/classification/*genetics ; Genome, Helminth ; Geography ; Humans ; Phylogeny ; Wolbachia/genetics ; }, abstract = {Filarial parasites are members of the Phylum Nemata that comprise several species of medical and veterinary importance. Among the human diseases caused by members of this group of nematodes are river blindness and lymphatic filariasis, which afflict millions of people in the tropics. These diseases not only have an impact on the health of the people affected but also bear a great socioeconomic burden. Despite their relevance, the systematics of the filarial parasites is not well understood yet, and additional molecular phylogenetic studies are required to comprehend the evolution of these parasites. Identifying the patterns of evolution of these parasites will be of relevance in preventing emerging zoonoses. The present review examines the information about the molecular systematics of filarial parasites available in the literature and evaluates the relevance of the different directions of future research. Furthermore, it is also intended to highlight the relevance of molecular systematic studies in the molecular epidemiology research area.}, } @article {pmid19536597, year = {2009}, author = {Coscrato, VE and Braz, AS and P Perondini, AL and Selivon, D and Marino, CL}, title = {Wolbachia in Anastrepha fruit flies (Diptera: Tephritidae).}, journal = {Current microbiology}, volume = {59}, number = {3}, pages = {295-301}, pmid = {19536597}, issn = {1432-0991}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Brazil ; Cluster Analysis ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Ecuador ; Female ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Tephritidae/*microbiology ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and cause a variety of reproductive abnormalities, such as cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing, and host feminization. In this study, we used three sets of Wolbachia-specific primers (16S rDNA, ftsZ, and wsp) in conjunction with the polymerase chain reaction (PCR), cloning and sequencing to study the infection of fruit flies (Anastrepha spp. and Ceratitis capitata) by Wolbachia. The flies were collected at several localities in Brazil and at Guayaquil, Ecuador. All of the fruit flies studied were infected with Wolbachia supergroup A, in agreement with the high prevalence of this group in South America. Phylogenetic analysis showed that the wsp gene was the most sensitive gene for studying the relationships among Wolbachia strains. The Wolbachia sequences detected in these fruit flies were similar to those such as wMel reported for other fruit flies. These results show that the infection of Anastrepha fruit flies by Wolbachia is much more widespread than previously thought.}, } @article {pmid19529776, year = {2009}, author = {Barquero-Calvo, E and Conde-Alvarez, R and Chacón-Díaz, C and Quesada-Lobo, L and Martirosyan, A and Guzmán-Verri, C and Iriarte, M and Mancek-Keber, M and Jerala, R and Gorvel, JP and Moriyón, I and Moreno, E and Chaves-Olarte, E}, title = {The differential interaction of Brucella and ochrobactrum with innate immunity reveals traits related to the evolution of stealthy pathogens.}, journal = {PloS one}, volume = {4}, number = {6}, pages = {e5893}, pmid = {19529776}, issn = {1932-6203}, mesh = {Animals ; Brucella abortus/genetics/*metabolism/*pathogenicity ; Dendritic Cells/microbiology ; Female ; *Immunity, Innate ; Inflammation ; Lipopolysaccharides/metabolism ; Macrophages/microbiology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Knockout ; NF-kappa B/metabolism ; Ochrobactrum/genetics/*metabolism/*pathogenicity ; *Virulence ; }, abstract = {BACKGROUND: During evolution, innate immunity has been tuned to recognize pathogen-associated molecular patterns. However, some alpha-Proteobacteria are stealthy intracellular pathogens not readily detected by this system. Brucella members follow this strategy and are highly virulent, but other Brucellaceae like Ochrobactrum are rhizosphere inhabitants and only opportunistic pathogens. To gain insight into the emergence of the stealthy strategy, we compared these two phylogenetically close but biologically divergent bacteria.

In contrast to Brucella abortus, Ochrobactrum anthropi did not replicate within professional and non-professional phagocytes and, whereas neutrophils had a limited action on B. abortus, they were essential to control O. anthropi infections. O. anthropi triggered proinflammatory responses markedly lower than Salmonella enterica but higher than B. abortus. In macrophages and dendritic cells, the corresponding lipopolysaccharides reproduced these grades of activation, and binding of O. anthropi lipopolysaccharide to the TLR4 co-receptor MD-2 and NF-kappaB induction laid between those of B. abortus and enteric bacteria lipopolysaccharides. These differences correlate with reported variations in lipopolysaccharide core sugars, sensitivity to bactericidal peptides and outer membrane permeability.

CONCLUSIONS/SIGNIFICANCE: The results suggest that Brucellaceae ancestors carried molecules not readily recognized by innate immunity, so that non-drastic variations led to the emergence of stealthy intracellular parasites. They also suggest that some critical envelope properties, like selective permeability, are profoundly altered upon modification of pathogen-associated molecular patterns, and that this represents a further adaptation to the host. It is proposed that this adaptive trend is relevant in other intracellular alpha-Proteobacteria like Bartonella, Rickettsia, Anaplasma, Ehrlichia and Wolbachia.}, } @article {pmid19523069, year = {2009}, author = {Machtelinckx, T and Van Leeuwen, T and Vanholme, B and Gehesquière, B and Dermauw, W and Vandekerkhove, B and Gheysen, G and De Clercq, P}, title = {Wolbachia induces strong cytoplasmic incompatibility in the predatory bug Macrolophus pygmaeus.}, journal = {Insect molecular biology}, volume = {18}, number = {3}, pages = {373-381}, doi = {10.1111/j.1365-2583.2009.00877.x}, pmid = {19523069}, issn = {1365-2583}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Female ; Fluorescent Antibody Technique ; Heteroptera/*microbiology/physiology ; Male ; Phylogeny ; Polymerase Chain Reaction ; Reproduction ; *Symbiosis ; Tetracycline ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Macrolophus pygmaeus is a heteropteran predator that is widely used in European glasshouses for the biological control of whiteflies, aphids, thrips and spider mites. We have demonstrated that the insect is infected with the endosymbiotic bacterium Wolbachia pipientis. Several gene fragments of the endosymbiont were sequenced and subsequently used for phylogenetic analysis, revealing that it belongs to the Wolbachia supergroup B. The endosymbiont was visualized within the ovarioles using immunolocalization. Tetracycline treatments were used to cure M. pygmaeus from its infection. Although a completely cured line could not be obtained by this approach, the application of a constant antibiotic pressure over 13 generations resulted in a line with a significantly reduced Wolbachia concentration. Crosses performed with this tetracycline-treated line revealed that the endosymbiont causes severe cytoplasmic incompatibility. This is the first report of a reproductive effect induced by Wolbachia in an economically important heteropteran predator that may have vital implications for its commercial production and use in biological control.}, } @article {pmid19519634, year = {2009}, author = {Dannowski, J and Flor, M and Telschow, A and Hammerstein, P}, title = {The effect of sibmating on the infection dynamics of male-killing bacteria.}, journal = {Evolution; international journal of organic evolution}, volume = {63}, number = {10}, pages = {2525-2534}, doi = {10.1111/j.1558-5646.2009.00749.x}, pmid = {19519634}, issn = {1558-5646}, mesh = {Animals ; Female ; Insecta/*microbiology ; Male ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Male-killing (MK) bacteria are vertically transmitted endosymbionts that selectively kill the male offspring of their hosts. Simple mathematical models describe the infection dynamics using two parameters, the bacterial transmission rate and a fitness compensation for siblings of killed males. These models cannot explain two phenomena that have been observed in nature: the persistence of extreme MK causing all-female broods, and the coexistence of two different strains of MK bacteria in the same host population. In the present study, we extend the simple MK models and investigate theoretically the effects of sibmating on the infection dynamics. We demonstrate analytically that, in general, sibmating reduces MK prevalence, and can even cause its extinction. As a special case of this finding, we show that sibmating allows a stable coexistence between no infection and extreme MK. Furthermore, we performed computer simulations and showed that, depending on male mating capacity, a stable coexistence of two strains is possible if sibmating occurs but is below a threshold. The results suggest that sibmating might be an important factor for understanding the infection dynamics of MK bacteria.}, } @article {pmid19513092, year = {2009}, author = {Kremer, N and Charif, D and Henri, H and Bataille, M and Prévost, G and Kraaijeveld, K and Vavre, F}, title = {A new case of Wolbachia dependence in the genus Asobara: evidence for parthenogenesis induction in Asobara japonica.}, journal = {Heredity}, volume = {103}, number = {3}, pages = {248-256}, doi = {10.1038/hdy.2009.63}, pmid = {19513092}, issn = {1365-2540}, mesh = {Animals ; Female ; *Host-Pathogen Interactions ; Male ; Molecular Sequence Data ; *Parthenogenesis ; Phylogeny ; Sexual Behavior, Animal ; Species Specificity ; Symbiosis ; Wasps/microbiology/*physiology ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Wolbachia is a maternally inherited bacterium that is widely distributed among arthropods, in which it manipulates the reproduction of its hosts. Although generally facultative for its hosts, Wolbachia has recently become obligatory in Asobara tabida (Hymenoptera: Braconidae) in which it is required for the completion of oogenesis. Here, we describe a new Wolbachia strain (wAjap) that is associated with the genus Asobara and infects Asobara japonica. wAjap was detected in all female-biased populations of A. japonica found in the main islands of Japan, but not in the arrhenotokous populations from the southern islands. Using phylogenetic analyses based on multi-locus sequence typing (MLST), we show that this strain is closely related to wAtab3 (the strain required for oogenesis in A. tabida), even though they differ on Wolbachia surface protein (WSP) and WO phage sequences. Using antibiotic treatments, we show that cured thelytokous females are not dependent on Wolbachia for oogenesis. However, they produced only sons, showing that wAjap induces thelytokous parthenogenesis. Analyses of mating behavior and offspring production of individuals from Wolbachia-infected populations showed that while males were still sexually functional, females no longer attract males, making Wolbachia an obligate partner for daughter production in thelytokous populations. The fact that Wolbachia has become independently obligatory in two species of the same genus tends to show that dependence evolution can be common and swift, although no clear benefit for the parasitoid can be attributed to this dependence. Although dependence should lead to co-divergence between Wolbachia and its hosts, the very few cases of co-speciation observed in host-Wolbachia associations question the stability of these obligatory associations.}, } @article {pmid19513090, year = {2009}, author = {Hoffmann, AA}, title = {Essential but unhelpful wasp Wolbachia.}, journal = {Heredity}, volume = {103}, number = {3}, pages = {194-195}, doi = {10.1038/hdy.2009.50}, pmid = {19513090}, issn = {1365-2540}, mesh = {Animals ; Female ; Species Specificity ; Wasps/genetics/*microbiology/physiology ; Wolbachia/genetics/*physiology ; }, } @article {pmid19505052, year = {2009}, author = {Voronin, DA and Bochernikov, AM and Baricheva, EM and Zakharov, IK and Kiseleva, EV}, title = {[Influence of Drosophila melanogaster genotype on biological effects of endocymbiont Wolbachia (stamm wMelPop)].}, journal = {Tsitologiia}, volume = {51}, number = {4}, pages = {335-345}, pmid = {19505052}, issn = {0041-3771}, mesh = {Animals ; DNA-Binding Proteins/genetics ; Drosophila Proteins/genetics ; Drosophila melanogaster/genetics/*microbiology/*physiology/ultrastructure ; Embryo, Nonmammalian/microbiology/ultrastructure ; Female ; Genome, Insect ; Longevity/genetics ; Mutation ; Reproduction ; Species Specificity ; *Symbiosis ; Transcription Factors/genetics ; Wolbachia/*physiology/ultrastructure ; }, abstract = {Comparative analyses of symbiotic bacteria Wolbachia (stamm wMelPop reducing lifespan of flies) morphology in normal and mutant strains of Drosophila melanogaster as well as the influence of Wolbachia on the host cell ultrastructure have been done. Wolbachia infected D. melanogaster mutation strains Trithorax-like -- Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+ have been received by special flies crossing. Uninfected strain D. melanogaster white-1118 (w1118) have been obtained by antibiotic treatment of initially infected strain D. melanogaster [w]w1118. Complex of different methods and approaches let to investigate for the first time the morphology of cell structure before and after bacterial infection of insects and to value the bacterial presence effect on flies viability and reproduction of normal and mutant flies. Morphology af cytoplasmic compartments in early embryos and eggs layed by infected and uninfecyed females Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+ have been analyzed. Electron microscopy has shown that D. melanogaster embryos contain typical Wolbachia contacting with different host organelles that verify preservation of their functional activity. Atificial mitochondria and Wolbachia (wMelPop) of unusual morphology with defective bacterial membranes have been visualised in D. melanogaster [w]Trl362/TM3, Sb1 Ser y+. Wolbachia presence in ovarium cells from strains [w]Trl362/TM3, Sb1 Ser y+ and [w]Trlen82/TM3, Sb1 Ser y+ did not influence on eggs quantity layed by females. We have demonstrated for the first time that lifespan of infected and uninfected strains: D. melanogaster Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+ were similar. However the lifespan of imago from strain [w]w1118 was lower in comparison to those from strains Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+. It suggests that either chromosomal balancer TM3 or Trl mutation play an importance role in host-symbiotic relationship. Next experiments have revealed that lifespan of homozygotic flies decreased essentially and was close to lifespan of strain [w]w1118. Data obtained confirm that chromosomal balancer TM3 can affect on symbiont-host relationship.}, } @article {pmid19504759, year = {2009}, author = {Haegeman, A and Vanholme, B and Jacob, J and Vandekerckhove, TT and Claeys, M and Borgonie, G and Gheysen, G}, title = {An endosymbiotic bacterium in a plant-parasitic nematode: member of a new Wolbachia supergroup.}, journal = {International journal for parasitology}, volume = {39}, number = {9}, pages = {1045-1054}, doi = {10.1016/j.ijpara.2009.01.006}, pmid = {19504759}, issn = {1879-0135}, mesh = {Animals ; Arthropods/*genetics/parasitology ; Gene Expression Profiling ; Genes, Helminth/*genetics ; Host-Parasite Interactions ; Molecular Sequence Data ; Nematoda/*genetics ; Phylogeny ; Sequence Analysis, DNA ; Symbiosis/*genetics ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia is an endosymbiotic bacterium widely present in arthropods and animal-parasitic nematodes. Despite previous efforts, it has never been identified in plant-parasitic nematodes. Random sequencing of genes expressed by the burrowing nematode Radopholus similis resulted in several sequences with similarity to Wolbachia genes. The presence of a Wolbachia-like endosymbiont in this plant-parasitic nematode was investigated using both morphological and molecular approaches. Transmission electronmicroscopy, fluorescent immunolocalisation and staining with DAPI confirmed the presence of the endosymbiont within the reproductive tract of female adults. 16S rDNA, ftsZ and groEL gene sequences showed that the endosymbiont of R. similis is distantly related to the known Wolbachia supergroups. Finally, based on our initial success in finding sequences of this endosymbiont by screening an expressed sequence tag (EST) dataset, all nematode ESTs were mined for Wolbachia-like sequences. Although the retained sequences belonged to six different nematode species, R. similis was the only plant-parasitic nematode with traces of Wolbachia. Based on our phylogenetic study and the current literature we designate the endosymbiont of R. similis to a new supergroup (supergroup I) rather than considering it as a new species. Although its role remains unknown, the endosymbiont was found in all individuals tested, pointing towards an essential function of the bacteria.}, } @article {pmid19500146, year = {2009}, author = {Carrington, LB and Leslie, J and Weeks, AR and Hoffmann, AA}, title = {The popcorn Wolbachia infection of Drosophila melanogaster: can selection alter Wolbachia longevity effects?.}, journal = {Evolution; international journal of organic evolution}, volume = {63}, number = {10}, pages = {2648-2657}, doi = {10.1111/j.1558-5646.2009.00745.x}, pmid = {19500146}, issn = {1558-5646}, mesh = {Animals ; Drosophila melanogaster/genetics/*microbiology ; Longevity/*genetics ; *Selection, Genetic ; Wolbachia/*physiology ; }, abstract = {Wolbachia popcorn (wMelPop), a life-shortening strain of Wolbachia, has been proposed as an agent for suppressing transmission of dengue fever following infection of the vectoring mosquito Aedes aegypti. However, evolutionary changes in the host and Wolbachia genomes might attenuate any life span effects mediated by wMelPop. Here we test for attenuation by selecting strains of Drosophila melanogaster infected with wMelPop for early and late reproduction in three independent outcrossed populations. Selection caused divergence among the lines in longevity. This divergence was mostly associated with the host genetic background rather than the Wolbachia infection, although there were also interactions between the host and Wolbachia genomes. Development time, viability, and productivity were not altered by selection. The implications of these results are discussed in light of the intended use of wMelPop for suppressing disease transmission.}, } @article {pmid19496412, year = {2009}, author = {Gavotte, L and Mercer, DR and Vandyke, R and Mains, JW and Dobson, SL}, title = {Wolbachia infection and resource competition effects on immature Aedes albopictus (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {46}, number = {3}, pages = {451-459}, pmid = {19496412}, issn = {0022-2585}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; AI-51533/AI/NIAID NIH HHS/United States ; R01 AI067434-01A1/AI/NIAID NIH HHS/United States ; R01 AI051533-01A1/AI/NIAID NIH HHS/United States ; AI-67434/AI/NIAID NIH HHS/United States ; R01 AI067434/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/growth & development/*microbiology/physiology ; Animals ; *Competitive Behavior ; Feeding Behavior ; Female ; Larva/growth & development/microbiology/physiology ; Male ; Sex Factors ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis Hertig and Wolbach (Rickettsiales: Rickettsiaceae) are intracellular alpha-proteobacteria that occur naturally in Aedes albopictus (Skuse) (Diptera: Culicidae) and numerous other invertebrates. These endosymbionts can invade host populations by manipulating host reproduction. Wolbachia infections have been shown to impart both costs and benefits to hosts in terms of development, survival, and fecundity. Here, we monitor intraspecific competition among independent cohorts of infected or uninfected larvae. Levels of competition are manipulated by varying initial larval densities and food levels. Although larval density is observed to have major impacts on immature survivorship, sex ratio of eclosing adults, and developmental rates, the Wolbachia infection status had minimal impact on male immatures and no effect on immature females under these experimental conditions. Female and male immatures were observed to respond differently to competitive pressure, with the functional relationships of females and males consistent with scramble and contest competition, respectively. The results are discussed in relation to the evolution of naturally occurring Wolbachia infections in Ae. albopictus (i.e., natural population replacement events) and public health strategies that propose the manipulation of Wolbachia infections in Ae. albopictus populations.}, } @article {pmid19492088, year = {2009}, author = {Watts, T and Haselkorn, TS and Moran, NA and Markow, TA}, title = {Variable incidence of Spiroplasma infections in natural populations of Drosophila species.}, journal = {PloS one}, volume = {4}, number = {5}, pages = {e5703}, pmid = {19492088}, issn = {1932-6203}, mesh = {Animals ; Drosophila/*microbiology ; Geography ; Gram-Negative Bacterial Infections/*epidemiology ; Incidence ; Population Dynamics ; Spiroplasma/*physiology ; }, abstract = {Spiroplasma is widespread as a heritable bacterial symbiont in insects and some other invertebrates, in which it sometimes acts as a male-killer and causes female-biased sex ratios in hosts. Besides Wolbachia, it is the only heritable bacterium known from Drosophila, having been found in 16 of over 200 Drosophila species screened, based on samples of one or few individuals per species. To assess the extent to which Spiroplasma infection varies within and among species of Drosophila, intensive sampling consisting of 50-281 individuals per species was conducted for natural populations of 19 Drosophila species. Infection rates varied among species and among populations of the same species, and 12 of 19 species tested negative for all individuals. Spiroplasma infection never was fixed, and the highest infection rates were 60% in certain populations of D. hydei and 85% in certain populations of D. mojavensis. In infected species, infection rates were similar for males and females, indicating that these Spiroplasma infections do not confer a strong male-killing effect. These findings suggest that Spiroplasma has other effects on hosts that allow it to persist, and that environmental or host variation affects transmission or persistence leading to differences among populations in infection frequencies.}, } @article {pmid19490857, year = {2009}, author = {Marshall, JM}, title = {The effect of gene drive on containment of transgenic mosquitoes.}, journal = {Journal of theoretical biology}, volume = {258}, number = {2}, pages = {250-265}, doi = {10.1016/j.jtbi.2009.01.031}, pmid = {19490857}, issn = {1095-8541}, support = {1R56AI072549/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culicidae/*genetics/microbiology/physiology ; Genetic Engineering/methods ; Insect Control/methods ; Insect Vectors/*genetics ; Meiosis ; *Models, Genetic ; Organisms, Genetically Modified/*genetics ; Population Dynamics ; *Repetitive Sequences, Nucleic Acid ; Risk Management ; *Stochastic Processes ; Wolbachia/genetics ; }, abstract = {Mosquito-borne diseases such as malaria and dengue fever continue to be a major health problem through much of the world. Several new potential approaches to disease control utilize gene drive to spread anti-pathogen genes into the mosquito population. Prior to a release, these projects will require trials in outdoor cages from which transgenic mosquitoes may escape, albeit in small numbers. Most genes introduced in small numbers are very likely to be lost from the environment; however, gene drive mechanisms enhance the invasiveness of introduced genes. Consequently, introduced transgenes may be more likely to persist than ordinary genes following an accidental release. Here, we develop stochastic models to analyze the loss probabilities for several gene drive mechanisms, including homing endonuclease genes, transposable elements, Medea elements, the intracellular bacterium Wolbachia, engineered underdominance genes, and meiotic drive. We find that Medea and Wolbachia present the best compromise between invasiveness and containment for the six gene drive systems currently being considered for the control of mosquito-borne disease.}, } @article {pmid19489671, year = {2009}, author = {Hudson, AJ and Floate, KD}, title = {Further evidence for the absence of bacteria in horsehair worms (Nematomorpha: Gordiidae).}, journal = {The Journal of parasitology}, volume = {95}, number = {6}, pages = {1545-1547}, doi = {10.1645/GE-2145.1}, pmid = {19489671}, issn = {1937-2345}, mesh = {Animals ; Bacteria/genetics/*isolation & purification ; Consensus Sequence ; DNA, Bacterial/chemistry/isolation & purification ; DNA, Ribosomal/chemistry ; Electrophoresis, Agar Gel ; Gryllidae/*parasitology ; Helminths/genetics/*microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 28S/genetics ; Sequence Alignment ; }, abstract = {We used molecular techniques to characterize bacteria associated with the nematomorph Gordius robustus (Leidy). This worm is a parasite of the fall field cricket, Gryllus pennsylvanicus (Burmeister), which is infected with the symbiotic bacteria, Wolbachia. Because of this close association, our a priori expectation was that G. robustus may be similarly infected. However, results of denaturing gradient gel electrophoresis and sequencing of amplified 16S rDNA failed to detect any bacteria (symbiotic or non-symbiotic) in G. robustus. These unexpected findings suggest that G. robustus has no internal bacterial community and indicate that close association with a Wolbachia-infected host is insufficient for the transmission of bacteria from insect to nematomorph.}, } @article {pmid19486895, year = {2009}, author = {Pichon, S and Bouchon, D and Cordaux, R and Chen, L and Garrett, RA and Grève, P}, title = {Conservation of the Type IV secretion system throughout Wolbachia evolution.}, journal = {Biochemical and biophysical research communications}, volume = {385}, number = {4}, pages = {557-562}, doi = {10.1016/j.bbrc.2009.05.118}, pmid = {19486895}, issn = {1090-2104}, mesh = {Biological Transport/genetics ; DNA/*metabolism ; *Evolution, Molecular ; Gene Order ; Genome, Bacterial ; *Operon ; Protein Transport/genetics ; Proteins/*metabolism ; Wolbachia/*genetics/*metabolism ; }, abstract = {The Type IV Secretion System (T4SS) is an efficient pathway with which bacteria can mediate the transfer of DNA and/or proteins to eukaryotic cells. In Wolbachia pipientis, a maternally inherited obligate endosymbiont of arthropods and nematodes, two operons of vir genes, virB3-B6 and virB8-D4, encoding a T4SS were previously identified and characterized at two separate genomic loci. Using the largest data set of Wolbachia strains studied so far, we show that vir gene sequence and organization are strictly conserved among 37 Wolbachia strains inducing various phenotypes such as cytoplasmic incompatibility, feminization, or oogenesis in their arthropod hosts. In sharp contrast, extensive variation of genomic sequences flanking the virB8-D4 operon suggested its distinct location among Wolbachia genomes. Long term conservation of the T4SS may imply maintenance of a functional effector translocation system in Wolbachia, thereby suggesting the importance for the T4SS in Wolbachia biology and survival inside host cells.}, } @article {pmid19485799, year = {2009}, author = {Bockarie, MJ and Taylor, MJ and Gyapong, JO}, title = {Current practices in the management of lymphatic filariasis.}, journal = {Expert review of anti-infective therapy}, volume = {7}, number = {5}, pages = {595-605}, doi = {10.1586/eri.09.36}, pmid = {19485799}, issn = {1744-8336}, mesh = {Animals ; Brugia/drug effects/microbiology ; Culex/parasitology ; Delivery of Health Care ; Elephantiasis, Filarial/*drug therapy/parasitology/*prevention & control/transmission ; Filaricides/*therapeutic use ; Humans ; Insect Vectors/parasitology ; Microfilariae/drug effects ; Wolbachia/drug effects ; Wuchereria bancrofti/drug effects/microbiology ; }, abstract = {Lymphatic filariasis is a major cause of acute and chronic morbidity in 81 countries. The availability of safe treatment regimens along with rapid diagnostic tools resulted in a global program to eliminate the disease. The two main objectives of the global elimination program are to interrupt transmission of the parasites and to provide care for those with the disease. The strategy for transmission interruption is preventive chemotherapy through mass drug administration. This article reviews the current treatment regimens for lymphatic filariasis and discusses the challenges posed by co-endemicity with other diseases. The role of integrated vector management as a supplementary strategy for mass drug administration and new strategies for treatment and morbidity control through antibiotic targeting of the Wolbachia endosymbionts are also discussed.}, } @article {pmid19479746, year = {2009}, author = {Ghanim, M and Kontsedalov, S}, title = {Susceptibility to insecticides in the Q biotype of Bemisia tabaci is correlated with bacterial symbiont densities.}, journal = {Pest management science}, volume = {65}, number = {9}, pages = {939-942}, doi = {10.1002/ps.1795}, pmid = {19479746}, issn = {1526-4998}, mesh = {Animals ; Female ; Gram-Negative Bacteria/genetics/*physiology ; Hemiptera/*drug effects/*microbiology/physiology ; Insecticides/*pharmacology ; *Symbiosis ; }, abstract = {BACKGROUND: The presence of symbiotic microorganisms may influence an insect's ability to tolerate natural and artificial stress agents such as insecticides. The authors have previously shown that Rickettsia in the B biotype of the whitefly Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) increases this insect's susceptibility to several insecticidal compounds. This communication reports a comparison of the susceptibilities of three isofemale strains of the Q biotype of B. tabaci harbouring different bacterial complements to major insecticides from different chemical groups: one strain harboured only Arsenophonus, one harboured Rickettsia and Arsenophonus and one harboured Arsenophonus and Wolbachia.

RESULTS: The presence of different symbiont combinations in the three strains had a significant influence on their susceptibility to most of the insecticides tested. Thiamethoxam, imidacloprid, pyriproxyfen and spiromesifen had a significant influence on strains that had the double infections Rickettsia-Arsenophonus and Wolbachia-Arsenophonus, which also carried higher amounts of symbionts as assessed by quantitative real-time PCR. No significant differences in mortality rates were observed when the tested strains were treated with diafenthiuron.

CONCLUSION: The results suggest a correlation between the presence of high bacterial densities in B. tabaci and the insect's ability to detoxify toxic compounds such as insecticides.}, } @article {pmid19478258, year = {2009}, author = {Debrah, AY and Mand, S and Marfo-Debrekyei, Y and Batsa, L and Pfarr, K and Lawson, B and Taylor, M and Adjei, O and Hoerauf, A}, title = {Reduction in levels of plasma vascular endothelial growth factor-A and improvement in hydrocele patients by targeting endosymbiotic Wolbachia sp. in Wuchereria bancrofti with doxycycline.}, journal = {The American journal of tropical medicine and hygiene}, volume = {80}, number = {6}, pages = {956-963}, pmid = {19478258}, issn = {1476-1645}, mesh = {Adolescent ; Adult ; Albendazole/therapeutic use ; Animals ; Anti-Bacterial Agents/pharmacology/*therapeutic use ; Double-Blind Method ; Doxycycline/pharmacology/*therapeutic use ; Filariasis/complications/drug therapy ; Filaricides/therapeutic use ; Ghana/epidemiology ; Humans ; Ivermectin/therapeutic use ; Male ; Middle Aged ; Testicular Hydrocele/blood/*drug therapy/epidemiology/etiology ; Vascular Endothelial Growth Factor A/*blood ; Wolbachia/*drug effects ; Wuchereria bancrofti/*microbiology ; Young Adult ; }, abstract = {The treatment for hydrocele is expensive, invasive surgery-hydrocelectomy. A drug that could prevent or improve this condition could replace or supplement hydrocelectomy. In Ghana, 42 hydrocele patients participated in a double-blind, placebo-controlled trial of a six-week regimen of doxycycline, 200 mg/day. Four months after doxycycline treatment, patients received 150 mug/kg of ivermectin and 400 mg of albendazole, which is used for mass chemotherapy in this area. Patients were monitored for levels of Wolbachia sp., microfilaremia, antigenemia, plasma levels of vascular endothelial growth factor-A (VEGF-A) and stage/size of the hydrocele. Wolbachia sp. loads/microfilaria, microfilaremia, and antigenemia were significantly reduced in the doxycycline-treated patients compared with the placebo group. The mean plasma levels of VEGF-A were decreased significantly in the doxycycline-treated patients who had active infection. This finding preceded the reduction of the stage of hydrocele. A six-week regimen of doxycycline treatment against filariasis showed amelioration of pathologic conditions of hydrocele patients with active infection.}, } @article {pmid19458089, year = {2009}, author = {Turner, JD and Langley, RS and Johnston, KL and Gentil, K and Ford, L and Wu, B and Graham, M and Sharpley, F and Slatko, B and Pearlman, E and Taylor, MJ}, title = {Wolbachia lipoprotein stimulates innate and adaptive immunity through Toll-like receptors 2 and 6 to induce disease manifestations of filariasis.}, journal = {The Journal of biological chemistry}, volume = {284}, number = {33}, pages = {22364-22378}, pmid = {19458089}, issn = {0021-9258}, support = {P30 EY011373/EY/NEI NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; EY11373/EY/NEI NIH HHS/United States ; R01 EY010320/EY/NEI NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Brugia/*metabolism ; Cell Line ; Dose-Response Relationship, Drug ; Female ; Filariasis/*metabolism ; Humans ; *Immune System ; Lipoproteins/*metabolism ; Mice ; Mice, Transgenic ; Models, Biological ; Toll-Like Receptor 2/*metabolism ; Toll-Like Receptor 6/*metabolism ; Wolbachia/*metabolism ; }, abstract = {Wolbachia endosymbiotic bacteria have been implicated in the inflammatory pathogenesis of filariasis. Inflammation induced by Brugia malayi female worm extract (BMFE) is dependent on Toll-like receptors 2 and 6 (TLR2/6) with only a partial requirement for TLR1. Removal of Wolbachia, lipids, or proteins eliminates all inflammatory activity. Wolbachia bacteria contain the lipoprotein biosynthesis genes Ltg and LspA but not Lnt, suggesting Wolbachia proteins cannot be triacylated, accounting for recognition by TLR2/6. Lipoprotein databases revealed 3-11 potential lipoproteins from Wolbachia. Peptidoglycan-associated lipoprotein (PAL) and Type IV secretion system-VirB6 were consistently predicted, and B. malayi Wolbachia PAL (wBmPAL) was selected for functional characterization. Diacylated 20-mer peptides of wBmPAL (Diacyl Wolbachia lipopeptide (Diacyl WoLP)) showed a near identical TLR2/6 and TLR2/1 usage compared with BMFE and bound directly to TLR2. Diacyl WoLP induced systemic tumor necrosis factor-alpha and neutrophil-mediated keratitis in mice. Diacyl WoLP activated monocytes induce up-regulation of gp38 on human lymphatic endothelial cells and induced dendritic cell maturation and activation. Dendritic cells primed with BMFE generated a non-polarized Th1/Th2 CD4+ T cell profile, whereas priming with Wolbachia depleted extracts (following tetracycline treatment; BMFEtet) polarized to a Th2 profile that could be reversed by reconstitution with Diacyl WoLP. BMFE generated IgG1 and IgG2c antibody responses, whereas BMFEtet or inoculation of TLR2 or MyD88-/- mice produced defective IgG2c responses. Thus, in addition to innate inflammatory activation, Wolbachia lipoproteins drive interferon-gamma-dependent CD4+ T cell polarization and antibody switching.}, } @article {pmid19452170, year = {2009}, author = {Korten, S and Büttner, DW and Schmetz, C and Hoerauf, A and Mand, S and Brattig, N}, title = {The nematode parasite Onchocerca volvulus generates the transforming growth factor-beta (TGF-beta).}, journal = {Parasitology research}, volume = {105}, number = {3}, pages = {731-741}, pmid = {19452170}, issn = {1432-1955}, mesh = {Animals ; Antibodies, Helminth/metabolism ; Brugia malayi/chemistry ; Dirofilaria/chemistry ; Epithelium/chemistry ; Goats ; Humans ; Immunohistochemistry ; Larva/chemistry/physiology ; Microscopy ; Microscopy, Immunoelectron ; Muscles/chemistry ; Onchocerca volvulus/chemistry/*physiology ; Subcutaneous Tissue/chemistry ; Transforming Growth Factor beta/*biosynthesis ; Wuchereria bancrofti/chemistry ; }, abstract = {Transforming growth factor-beta (TGF-beta) is a highly conserved cytokine that has a well-known regulatory role in immunity, but also in organ development of most animal species including helminths. Homologous tgf-b genes and mRNA have been detected in the filaria Brugia malayi. The in situ protein expression is unknown for filariae. Therefore, we examined several filariae for the expression and localization of latent (stable) TGF-beta in adult and larval stages. A specific goat anti-human latency associated protein (LAP, TGF-beta 1) antibody, purified by affinity chromatography, was used for light and electron microscopic immunohistochemistry. Adult Onchocerca volvulus, Onchocerca gibsoni, Onchocerca ochengi, Onchocerca armillata, Onchocerca fasciata, Onchocerca flexuosa, Wuchereria bancrofti, Dirofilaria sp., B. malayi, and infective larvae of W. bancrofti reacted with the antibody. Labeling of worm tissues varied between negative and all degrees of positive reactions. Latent TGF-beta was strongly expressed adjacent to the cell membranes of the hypodermis, epithelia, and muscles and adjacent to many nuclei in all organs. TGF-beta was well expressed in worms without Wolbachia endobacteria eliminated by doxycycline treatment. Pleomorphic neoplasms in O. volvulus were also labeled. We conclude that latent TGF-beta protein is expressed by filariae independently of Wolbachia, possibly regulating worm tissue homeostasis.}, } @article {pmid19450730, year = {2009}, author = {Oleaga, A and Pérez-Sánchez, R and Pagés, E and Marcos-Atxutegi, C and Simón, F}, title = {Identification of immunoreactive proteins from the dog heartworm (Dirofilaria immitis) differentially recognized by the sera from dogs with patent or occult infections.}, journal = {Molecular and biochemical parasitology}, volume = {166}, number = {2}, pages = {134-141}, doi = {10.1016/j.molbiopara.2009.03.005}, pmid = {19450730}, issn = {1872-9428}, mesh = {Amino Acid Sequence ; Animals ; Antibodies, Helminth/blood/*immunology ; Dirofilaria immitis/chemistry/*immunology ; Dirofilariasis/*immunology/parasitology ; Dog Diseases/*immunology/parasitology ; Dogs ; Electrophoresis, Gel, Two-Dimensional ; Helminth Proteins/chemistry/*immunology ; Molecular Sequence Data ; Sequence Alignment ; }, abstract = {Heartworm disease caused by Dirofilaria immitis affects canine and feline hosts. Moreover, the parasite can infect humans, causing pulmonary dirofilariosis. Most affected dogs have patent infections with circulating microfilariae in peripheral blood, although infected dogs sometimes develop occult infections characterized by the absence of microfilariae. Microfilaremic infections (mf+) are associated with a predominant Th2-type immune response, whereas a Th1-type response predominates in amicrofilaremic infections (mf-), suggesting a role for this response in the suppression of circulating microfilariae. However, nothing is known about the molecules involved in the immune regulation of these infections. The objective of the present work was to identify the parasite proteins recognized differentially by the immune response of dogs with patent or occult infections, using two-dimensional electrophoresis and mass spectrometry. Nineteen proteins of D. immitis were identified, of which 6 were immunoreactive against serum samples from both mf+ and mf- dogs, while another two groups of 6 and 7 different proteins were differentially recognized by sera from mf+ or mf- dogs, respectively. The results point to the existence of differential antigen recognition in patent and occult infections due to D. immitis. Several proteins that could be involved in the immune regulation of these infections are identified. Additionally, the findings seem to suggest that some antigens of D. immitis, together with Wolbachia antigens, could contribute to the stimulation of the Th1-type response.}, } @article {pmid19436325, year = {2009}, author = {Engelstädter, J and Telschow, A}, title = {Cytoplasmic incompatibility and host population structure.}, journal = {Heredity}, volume = {103}, number = {3}, pages = {196-207}, doi = {10.1038/hdy.2009.53}, pmid = {19436325}, issn = {1365-2540}, mesh = {Animals ; Cytoplasm/genetics/*microbiology ; Female ; Gene Flow ; *Host-Pathogen Interactions ; Insecta/genetics/*microbiology/physiology ; Male ; Models, Genetic ; Pest Control, Biological ; Wolbachia/physiology ; }, abstract = {Many arthropod species are infected by maternally inherited bacteria that induce cytoplasmic incompatibility (CI). CI causes embryonic mortality in offspring when infected males mate with either uninfected females or with females that are infected with a different strain of bacteria. Here, we review theoretical and empirical studies concerning the infection dynamics of CI-inducing bacteria, focusing in particular on the impact of the host population structure on the spread of CI. As different theoretical models have often produced divergent predictions with regard to issues such as the speed of CI spread and the stability of infection polymorphisms, we specifically aim to clarify how the various assumptions concerning population structure that underlie these models affect these predictions. We also discuss several implications of population structure, including the impact of CI on host gene flow reduction and speciation, the evolutionary dynamics of CI and strategies to control insect pest populations by means of CI-inducing microbes.}, } @article {pmid19430817, year = {2009}, author = {Datta, S and Maitra, S and Gayen, P and Sinha Babu, SP}, title = {Improved efficacy of tetracycline by acaciasides on Dirofilaria immitis.}, journal = {Parasitology research}, volume = {105}, number = {3}, pages = {697-702}, pmid = {19430817}, issn = {1432-1955}, mesh = {Animals ; Dirofilaria immitis/*drug effects ; Dirofilariasis/*drug therapy ; Dog Diseases/*drug therapy/parasitology ; Dogs ; Drug Synergism ; Female ; Filaricides/administration & dosage/*therapeutic use ; Male ; Saponins/administration & dosage/*therapeutic use ; Tetracycline/administration & dosage/*therapeutic use ; Treatment Outcome ; Triterpenes/administration & dosage/*therapeutic use ; }, abstract = {The discovery of Wolbachia, a bacterial endosymbiont that occurs in the filarial parasite and its sensitivity to tetracycline, has fostered a new initiative in the development of suitable antifilarial drugs. The present study is an attempt to investigate whether adding acaciasides (saponins from Acacia auriculiformis) to the standard dose of tetracycline would further improve the efficacy of tetracycline treatment against Dirofilaria immitis microfilariae in vivo. Treatment of microfilaremic adult dogs (body weight range 8-12 kg) with tetracycline at 10 mg/kg/day for 40 days resulted in 72% and 83% reduction in mf count on days 15 and 30, respectively, and the maximum reduction in mf count (91%) was achieved on day 75 post-treatment. However, treatment with tetracycline (10 mg/kg/day for 40 days) followed by acaciasides (10 mg/kg/day for 7 days) resulted in almost 100% clearance of mf at a faster rate on day 45 post-treatment and ensured long-term (until 4 months post-treatment) protection against microfilaremia. Data from polymerase chain reaction analysis reveals that compared to untreated dogs, in treated dogs, there was marked reduction in Wolbachia specific wsp markers in fast depleting mf population. The present data indicate that prior tetracycline treatment enhances microfilaricidal activity of saponins. This effect may be additive or synergistic as the worms are weakened by Wolbachia depletion, and these weakened microfilariae are possibly killed by the saponins.}, } @article {pmid19419989, year = {2009}, author = {Unckless, RL and Boelio, LM and Herren, JK and Jaenike, J}, title = {Wolbachia as populations within individual insects: causes and consequences of density variation in natural populations.}, journal = {Proceedings. Biological sciences}, volume = {276}, number = {1668}, pages = {2805-2811}, pmid = {19419989}, issn = {0962-8452}, mesh = {Aging ; Animals ; Drosophila/*microbiology ; Feeding Behavior ; Female ; Male ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {The population-level dynamics of maternally transmitted endosymbionts, including reproductive parasites, depends primarily on the fitness effects and transmission fidelity of these infections. Although experimental laboratory studies have shown that within-host endosymbiont density can affect both of these factors, the existence of such effects in natural populations has not yet been documented. Using quantitative PCR, we survey the density of male-killing Wolbachia in natural populations of Drosophila innubila females from the Chiricahua Mountains of Arizona. We find that there is substantial (20 000-fold) variation in Wolbachia density among wild flies and that within-host Wolbachia density is positively correlated with both the efficacy of male killing and maternal transmission fidelity. Mean Wolbachia density increases three- to five-fold from early to late in the season. This pattern suggests that Wolbachia density declines with fly age, a conclusion corroborated by a laboratory study of Wolbachia density as a function of age. Finally, we suggest three alternative hypotheses to account for the approximately lognormal distribution of Wolbachia density among wild flies.}, } @article {pmid19414524, year = {2009}, author = {Cordaux, R}, title = {Gene conversion maintains nonfunctional transposable elements in an obligate mutualistic endosymbiont.}, journal = {Molecular biology and evolution}, volume = {26}, number = {8}, pages = {1679-1682}, doi = {10.1093/molbev/msp093}, pmid = {19414524}, issn = {1537-1719}, mesh = {Animals ; *DNA Transposable Elements ; *Gene Conversion ; Nematoda/*microbiology/physiology ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {Long-term bacterial endosymbionts typically exhibit reduced genomes, lack genes encoding recombination functions and transposable elements, such as insertion sequences (ISs). In sharp contrast, I found that ISs constitute 2.4% of the genome of the obligate mutualistic endosymbiont Wolbachia wBm. Although no IS copy is transpositionally functional, I show that ISs persist in wBm because of frequent recombinational gene conversion (GC) homogenizing homologous IS sequences. These results not only indicate that there exists a functional recombination molecular machinery in wBm, but they also suggest that, by slowing down the rate of IS degradation and loss, GC may represent a major force influencing reductive evolution in wBm.}, } @article {pmid19411536, year = {2009}, author = {Evans, O and Caragata, EP and McMeniman, CJ and Woolfit, M and Green, DC and Williams, CR and Franklin, CE and O'Neill, SL and McGraw, EA}, title = {Increased locomotor activity and metabolism of Aedes aegypti infected with a life-shortening strain of Wolbachia pipientis.}, journal = {The Journal of experimental biology}, volume = {212}, number = {Pt 10}, pages = {1436-1441}, pmid = {19411536}, issn = {0022-0949}, mesh = {Aedes/*microbiology/physiology ; Animals ; Behavior, Animal ; Energy Metabolism/*physiology ; Female ; Host-Pathogen Interactions ; Male ; *Motor Activity ; Wolbachia/*physiology ; }, abstract = {A virulent strain of the obligate intracellular bacterium Wolbachia pipientis that shortens insect lifespan has recently been transinfected into the primary mosquito vector of dengue virus, Aedes aegypti L. The microbe's ability to shorten lifespan and spread through host populations under the action of cytoplasmic incompatibility means it has the potential to be used as a biocontrol agent to reduce dengue virus transmission. Wolbachia is present in many host tissues and may have local effects on diverse biological processes. In other insects, Wolbachia infections have been shown to alter locomotor activity and response time to food cues. In mosquitoes, locomotor performance relates to the location of mates, human hosts, resting sites and oviposition sites. We have therefore examined the effect of the virulent, life-shortening Wolbachia strain wMelPop on the locomotion of Ae. aegypti as they age and as the pathogenicity of the infection increases. In parallel experiments we also examined CO(2) production as a proxy for metabolic rate, to investigate a potential mechanistic explanation for any changes in locomotion. Contrary to expectation, we found that the infection increased activity and metabolic rate and that these effects were relatively consistent over the insect's lifespan. The results do not fit a standard model of bacterial pathogenesis in insects, and instead may reveal additional physiological changes induced by infection, such as either increased hunger or defects in the nervous system.}, } @article {pmid19389774, year = {2009}, author = {Ishmael, N and Hotopp, JCD and Ioannidis, P and Biber, S and Sakamoto, J and Siozios, S and Nene, V and Werren, J and Bourtzis, K and Bordenstein, SR and Tettelin, H}, title = {Extensive genomic diversity of closely related Wolbachia strains.}, journal = {Microbiology (Reading, England)}, volume = {155}, number = {Pt 7}, pages = {2211-2222}, pmid = {19389774}, issn = {1350-0872}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R01-GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Ankyrins/genetics ; Comparative Genomic Hybridization ; DNA, Bacterial/analysis/genetics ; Drosophila melanogaster/*microbiology ; Evolution, Molecular ; Gene Transfer, Horizontal ; Genes, Bacterial ; Genetic Variation ; Genome, Bacterial ; Interspersed Repetitive Sequences ; Species Specificity ; Wolbachia/*genetics ; }, abstract = {Using microarray-based comparative genome hybridization (mCGH), the genomic content of Wolbachia pipientis wMel from Drosophila melanogaster was compared to the closely related Wolbachia from D. innubila (wInn), D. santomea (wSan), and three strains from D. simulans (wAu, wRi, wSim). A large number of auxiliary genes are identified in these five strains, with most absent/divergent genes being unique to a given strain. Each strain caused an average of approximately 60 genes to be removed from the core genome. As such, these organisms do not appear to have the streamlined genomes expected of obligate intracellular bacteria. Prophage, hypothetical and ankyrin repeat genes are over-represented in the absent/divergent genes, with 21-87% of absent/divergent genes coming from prophage regions. The only wMel region absent/divergent in all five query strains is that containing WD_0509 to WD_0511, including a DNA mismatch repair protein MutL-2, a degenerate RNase, and a conserved hypothetical protein. A region flanked by the two portions of the WO-B prophage in wMel is found in four of the five Wolbachia strains as well as on a plasmid of a rickettsial endosymbiont of Ixodes scapularis, suggesting lateral gene transfer between these two obligate intracellular species. Overall, these insect-associated Wolbachia have highly mosaic genomes, with lateral gene transfer playing an important role in their diversity and evolution.}, } @article {pmid19384745, year = {2009}, author = {Martínez, P and Del Castillo, P and Bella, JL}, title = {Cytological detection of Wolbachia in squashed and paraffin embedded insect tissues.}, journal = {Biotechnic & histochemistry : official publication of the Biological Stain Commission}, volume = {84}, number = {6}, pages = {347-353}, doi = {10.3109/10520290902903381}, pmid = {19384745}, issn = {1473-7760}, mesh = {Animals ; Cytoplasm/*chemistry/genetics ; Insecta/*genetics ; Male ; Testis/*chemistry/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia localization in situ is essential for accurate analysis of the infection and its consequences. Whole cell hybridization is proposed as an easy and rapid method for detecting Wolbachia cells in paraffin embedded tissues or testis squashes of Chorthippus parallelus (Orthoptera). Wolbachia is found in whole gonads and other adjacent tissues. A higher bacterial density, however, is observed in ovarioles and testis. Small independent bacteria with strictly cytoplasmic distribution are displayed. Bacterial density differences among individuals are also revealed.}, } @article {pmid19383035, year = {2009}, author = {Wolfgang, A and Markus, R and Dimitrios, A and Christian, S}, title = {Evidence for low-titre infections in insect symbiosis: Wolbachia in the bark beetle Pityogenes chalcographus (Coleoptera, Scolytinae).}, journal = {Environmental microbiology}, volume = {11}, number = {8}, pages = {1923-1933}, doi = {10.1111/j.1462-2920.2009.01914.x}, pmid = {19383035}, issn = {1462-2920}, support = {P 21147/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics/metabolism ; DNA Fingerprinting ; Polymerase Chain Reaction/methods ; *Symbiosis ; Weevils/*microbiology ; Wolbachia/classification/*isolation & purification ; }, abstract = {Wolbachia are obligatory endosymbiotic alpha-proteobacteria found in many insect species. They are maternally transmitted and often exhibit reproductive phenotypes like cytoplasmic incompatibility. Pityogenes chalcographus is a bark beetle causing severe damage in spruce stands. Its European populations are divided into several mitochondrial clades separated by partial crossing barriers. In this study, we tested a large sample set covering the natural range of the beetle in Europe for the presence of Wolbachia and associations between infection pattern and mitotypes using a highly sensitive nested PCR technique. 35.5% of the individuals were infected with the endosymbiont and two distinct strains were identified. Both strains occur in low titre not accessible by conventional detection methods. The infections are present all over Europe, unlikely to cause the partial crossing barriers in this host and uncoupled from mitochondrial clades. This pattern is indicative for populations evolving towards endosymbiont loss and for repeated intraspecific horizontal transfer of Wolbachia. Alternatively, the low-titre infections found in P. chalcographus are yet another example for Wolbachia that can persist in host species at low densities and frequencies.}, } @article {pmid19379544, year = {2009}, author = {Specht, S and Wanji, S}, title = {New insights into the biology of filarial infections.}, journal = {Journal of helminthology}, volume = {83}, number = {2}, pages = {199-202}, doi = {10.1017/S0022149X09344922}, pmid = {19379544}, issn = {1475-2697}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Elephantiasis, Filarial/drug therapy/*prevention & control ; Filaricides/*therapeutic use ; Humans ; Onchocerciasis/drug therapy/*prevention & control ; }, abstract = {Recent successes in the control of lymphatic filariasis and onchocerciasis need continuing research in order to sustain the achievements and to develop further tools to tackle the new questions that arise when only reduced infection prevalences prevail. In this regard, in a symposium held at the Xth European Multicolloquium of Parasitology (August 2008, Paris) questions such as the impact of filarial immunosuppression, and its lack following filarial control, on the outcome of co-infections were addressed, as were new approaches to treatment with promising drugs such as moxidectin or the antibiotic chemotherapy against Wolbachia endosymbionts in filariae. In particular, longer treatment courses of doxycycline could be carried out by community-directed treatment at high coverage, thus potentially allowing its use in restricted areas with suboptimal responses to ivermectin against onchocerciasis, or in areas with co-infection by loiasis where onchocerciasis or lymphatic filariasis need to be controlled. New, more potent drugs, or eventually vaccines, will be of importance because in many vector-filarial parasite relationships worldwide, transmission efficacy increases with low numbers of ingested microfilariae, and since ivermectin may render treated hosts more susceptible to new infection.}, } @article {pmid19376923, year = {2009}, author = {Zouache, K and Voronin, D and Tran-Van, V and Mavingui, P}, title = {Composition of bacterial communities associated with natural and laboratory populations of Asobara tabida infected with Wolbachia.}, journal = {Applied and environmental microbiology}, volume = {75}, number = {11}, pages = {3755-3764}, pmid = {19376923}, issn = {1098-5336}, mesh = {Animals ; Bacteria/*classification/genetics/*isolation & purification ; *Biodiversity ; Cluster Analysis ; DNA Fingerprinting ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Electrophoresis, Polyacrylamide Gel ; France ; Gram-Negative Bacterial Infections/microbiology/*veterinary ; Hymenoptera/*microbiology ; Molecular Sequence Data ; Nucleic Acid Denaturation ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; }, abstract = {Asobara tabida wasps are fly endoparasitoids that naturally harbor three Wolbachia strains, which induce cytoplasmic incompatibility and control oogenesis. To investigate whether other bacteria play a role in wasp biology, we surveyed the bacterial communities of wild A. tabida populations originating from different regions of France and of laboratory colonies using PCR-denaturing gradient gel electrophoresis and culture methods. Proteobacteria and Firmicutes were found to be the main phyla represented in these populations. Among these were several cultured and uncultured representatives of the genera Acetobacter, Acidomonas, Bacillus, Brevibacillus, Duganella, Herbaspirillum, Pseudomonas, Staphylococcus, and Streptococcus. In addition to Wolbachia, wild individuals harbored Rickettsia, which tended to be lost when insects were reared in the laboratory. The antibiotic treatment used to generate wasp sublines singly infected with Wolbachia also affected the overall bacterial composition, with most fingerprint sequences being characteristic of the family Enterobacteriaceae. We also screened for potentially heritable endosymbionts by PCR and fluorescence in situ hybridization in stable laboratory lines, with only Wolbachia being consistently found in wasp ovaries.}, } @article {pmid19369264, year = {2009}, author = {Himler, AG and Caldera, EJ and Baer, BC and Fernández-Marín, H and Mueller, UG}, title = {No sex in fungus-farming ants or their crops.}, journal = {Proceedings. Biological sciences}, volume = {276}, number = {1667}, pages = {2611-2616}, pmid = {19369264}, issn = {0962-8452}, support = {K12 GM000708/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents ; Ants/microbiology/*physiology ; Female ; Fungi/*physiology ; Reproduction, Asexual/*physiology ; }, abstract = {Asexual reproduction imposes evolutionary handicaps on asexual species, rendering them prone to extinction, because asexual reproduction generates novel genotypes and purges deleterious mutations at lower rates than sexual reproduction. Here, we report the first case of complete asexuality in ants, the fungus-growing ant Mycocepurus smithii, where queens reproduce asexually but workers are sterile, which is doubly enigmatic because the clonal colonies of M. smithii also depend on clonal fungi for food. Degenerate female mating anatomy, extensive field and laboratory surveys, and DNA fingerprinting implicate complete asexuality in this widespread ant species. Maternally inherited bacteria (e.g. Wolbachia, Cardinium) and the fungal cultivars can be ruled out as agents inducing asexuality. M. smithii societies of clonal females provide a unique system to test theories of parent-offspring conflict and reproductive policing in social insects. Asexuality of both ant farmer and fungal crop challenges traditional views proposing that sexual farmer ants outpace coevolving sexual crop pathogens, and thus compensate for vulnerabilities of their asexual crops. Either the double asexuality of both farmer and crop may permit the host to fully exploit advantages of asexuality for unknown reasons or frequent switching between crops (symbiont reassociation) generates novel ant-fungus combinations, which may compensate for any evolutionary handicaps of asexuality in M. smithii.}, } @article {pmid19364731, year = {2009}, author = {Negri, I and Franchini, A and Gonella, E and Daffonchio, D and Mazzoglio, PJ and Mandrioli, M and Alma, A}, title = {Unravelling the Wolbachia evolutionary role: the reprogramming of the host genomic imprinting.}, journal = {Proceedings. Biological sciences}, volume = {276}, number = {1666}, pages = {2485-2491}, pmid = {19364731}, issn = {0962-8452}, mesh = {Animals ; *Biological Evolution ; DNA Methylation ; Epigenesis, Genetic ; Female ; *Genome, Insect ; *Genomic Imprinting ; Hemiptera/anatomy & histology/*microbiology ; Male ; Sex Determination Processes ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Environmental factors can induce significant epigenetic changes that may also be inherited by future generations. The maternally inherited symbiont of arthropods Wolbachia pipientis is an excellent candidate as an 'environmental' factor promoting trans-generational epigenetic changes: by establishing intimate relationships with germ-line cells, epigenetic effects of Wolbachia symbiosis would be manifested as a 'maternal effect', in which infection of the mother modulates the offspring phenotype. In the leafhopper Zyginidia pullula, Wolbachia feminizes genetic males, leaving them as intersexes. With the exception of male chitinous structures that are present in the last abdominal segment, feminized males display phenotypic features that are typical of females. These include ovaries that range from a typical histological architecture to an altered structure. Methylation-sensitive random amplification of polymorphic DNA profiles show that they possess a female genomic imprint. On the other hand, some rare feminized males bear testes instead of ovaries. These specimens possess a Wolbachia density approximately four orders of magnitude lower than feminized males with ovaries and maintain a male genome-methylation pattern. Our results indicate that Wolbachia infection disrupts male imprinting, which dramatically influences the expression of genes involved in sex differentiation and development, and the alteration occurs only if Wolbachia exceeds a density threshold. Thus, a new Wolbachia's role as an environmental evolutionary force, inducing epigenetic trans-generational changes, should now be considered.}, } @article {pmid19353951, year = {2009}, author = {Junker, K and Barbuto, M and Casiraghi, M and Martin, C and Uni, S and Boomker, J and Bain, O}, title = {Litomosa chiropterorum Ortlepp, 1932 (Nematoda: Filarioidea) from a South African miniopterid: redescription, Wolbachia screening and phylogenetic relationships with Litomosoides.}, journal = {Parasite (Paris, France)}, volume = {16}, number = {1}, pages = {43-50}, doi = {10.1051/parasite/2009161043}, pmid = {19353951}, issn = {1252-607X}, mesh = {Animals ; Chiroptera/*parasitology ; Female ; Filarioidea/*anatomy & histology/*classification/microbiology ; Male ; *Phylogeny ; Prevalence ; South Africa ; Wolbachia/isolation & purification ; }, abstract = {69 Miniopterus notalensis, type host of the onchocercid Litomosa chiropterorum, were collected in caves in the Western Province and Gauteng Province, South Africa. The prevalence of these filariae was about 50 %. The microfilaria is folded, as in other Litomosa and an area rugosa composed of cuticular bosses is present in the male posterior region. L. chiropterorum is close to the species parasitic in other Miniopterus spp. and some Rhinolophus spp. from Africa, Madagascar and Europe; it is unique with the expanded anterior extremity and the four cephalic submedian bosses. The molecular analysis of L. chiropterorum, the first done with Litomosa species from a bat, supports the hypothesis that Litomosa and Litomosoides, which have an exceptionally large buccal capsule in common, form a group in which Litomosa has a basal position. Interestingly, L. chiropterorum does not harbour Wolbachia, as proved with immunohistological staining and PCR screening using the 16S rDNA gene as target. This is contrary to L. westi from rodents and the majority of the Litomosoides species parasitic in bats or rodents. The absence of Wolbachia in a filarioid group considered ancient based on traditional and molecular approaches opens interesting scenarios on the evolution of the endosymbionts spread through filarial lineages.}, } @article {pmid19352421, year = {2009}, author = {Bennuru, S and Semnani, R and Meng, Z and Ribeiro, JM and Veenstra, TD and Nutman, TB}, title = {Brugia malayi excreted/secreted proteins at the host/parasite interface: stage- and gender-specific proteomic profiling.}, journal = {PLoS neglected tropical diseases}, volume = {3}, number = {4}, pages = {e410}, pmid = {19352421}, issn = {1935-2735}, support = {N01CO12400/CA/NCI NIH HHS/United States ; N01-CO-12400/CO/NCI NIH HHS/United States ; /ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*chemistry/*growth & development ; Chromatography, Liquid ; Erythrocytes ; Female ; Helminth Proteins/*metabolism ; *Host-Pathogen Interactions ; Male ; Mass Spectrometry ; Proteome/*analysis ; }, abstract = {Relatively little is known about the filarial proteins that interact with the human host. Although the filarial genome has recently been completed, protein profiles have been limited to only a few recombinants or purified proteins of interest. Here, we describe a large-scale proteomic analysis using microcapillary reverse-phase liquid chromatography-tandem-mass spectrometry to identify the excretory-secretory (ES) products of the L3, L3 to L4 molting ES, adult male, adult female, and microfilarial stages of the filarial parasite Brugia malayi. The analysis of the ES products from adult male, adult female, microfilariae (Mf), L3, and molting L3 larvae identified 852 proteins. Annotation suggests that the functional and component distribution was very similar across each of the stages studied; however, the Mf contributed a higher proportion to the total number of identified proteins than the other stages. Of the 852 proteins identified in the ES, only 229 had previous confirmatory expressed sequence tags (ESTs) in the available databases. Moreover, this analysis was able to confirm the presence of 274 "hypothetical" proteins inferred from gene prediction algorithms applied to the B. malayi (Bm) genome. Not surprisingly, the majority (160/274) of these "hypothetical" proteins were predicted to be secreted by Signal IP and/or SecretomeP 2.0 analysis. Of major interest is the abundance of previously characterized immunomodulatory proteins such as ES-62 (leucyl aminopeptidase), MIF-1, SERPIN, glutathione peroxidase, and galectin in the ES of microfilariae (and Mf-containing adult females) compared to the adult males. In addition, searching the ES protein spectra against the Wolbachia database resulted in the identification of 90 Wolbachia-specific proteins, most of which were metabolic enzymes that have not been shown to be immunogenic. This proteomic analysis extends our knowledge of the ES and provides insight into the host-parasite interaction.}, } @article {pmid19345018, year = {2009}, author = {Zhang, B and McGraw, E and Floate, KD and James, P and Jorgensen, W and Rothwell, J}, title = {Wolbachia infection in Australasian and North American populations of Haematobia irritans (Diptera: Muscidae).}, journal = {Veterinary parasitology}, volume = {162}, number = {3-4}, pages = {350-353}, doi = {10.1016/j.vetpar.2009.03.012}, pmid = {19345018}, issn = {0304-4017}, mesh = {Animals ; Australasia ; Host-Pathogen Interactions ; Muscidae/*microbiology ; North America ; Wolbachia/*physiology ; }, abstract = {Buffalo fly (Haematobia irritans exigua) is a major pest of beef and dairy cattle in northern Australia. Global warming is expected to increase the southern range of buffalo fly. Chemical control is compromised by resistance and may not be feasible in extensive production systems and there is rapidly growing market preference for beef and dairy products produced in low-chemical systems. Wolbachia are vertically transmitted intracellular bacteria that can profoundly influence host reproduction and fitness and are of increasing interest for use in biocontrol programs. To determine whether Australian flies are infected with Wolbachia, buffalo flies were collected from 12 cattle herds around Australia and assayed by standard PCR for the Wolbachia wsp gene. H. i. exigua from Indonesia and horn fly (H. i. irritans) from Canada were also tested. All H. i. exigua samples tested were negative for Wolbachia infection whereas a very strong signal for Wolbachia was obtained from H. i. irritans.}, } @article {pmid19343208, year = {2009}, author = {Brownlie, JC and Cass, BN and Riegler, M and Witsenburg, JJ and Iturbe-Ormaetxe, I and McGraw, EA and O'Neill, SL}, title = {Evidence for metabolic provisioning by a common invertebrate endosymbiont, Wolbachia pipientis, during periods of nutritional stress.}, journal = {PLoS pathogens}, volume = {5}, number = {4}, pages = {e1000368}, pmid = {19343208}, issn = {1553-7374}, mesh = {Analysis of Variance ; Animals ; Drosophila melanogaster/growth & development/*microbiology/physiology ; Female ; Fertility ; Iron/*metabolism ; Male ; Statistics, Nonparametric ; *Stress, Physiological ; *Symbiosis ; Wolbachia/metabolism/*physiology ; }, abstract = {Wolbachia are ubiquitous inherited endosymbionts of invertebrates that invade host populations by modifying host reproductive systems. However, some strains lack the ability to impose reproductive modification and yet are still capable of successfully invading host populations. To explain this paradox, theory predicts that such strains should provide a fitness benefit, but to date none has been detected. Recently completed genome sequences of different Wolbachia strains show that these bacteria may have the genetic machinery to influence iron utilization of hosts. Here we show that Wolbachia infection can confer a positive fecundity benefit for Drosophila melanogaster reared on iron-restricted or -overloaded diets. Furthermore, iron levels measured from field-collected flies indicated that nutritional conditions in the field were overall comparable to those of flies reared in the laboratory on restricted diets. These data suggest that Wolbachia may play a previously unrecognized role as nutritional mutualists in insects.}, } @article {pmid19341336, year = {2009}, author = {Wiwatanaratanabutr, I and Kittayapong, P and Caubet, Y and Bouchon, D}, title = {Molecular phylogeny of Wolbachia strains in arthropod hosts based on groE-homologous gene sequences.}, journal = {Zoological science}, volume = {26}, number = {2}, pages = {171-177}, doi = {10.2108/zsj.26.171}, pmid = {19341336}, issn = {0289-0003}, mesh = {Animals ; Arthropods/*microbiology ; Base Sequence ; Cloning, Molecular ; DNA, Bacterial/genetics ; Heat-Shock Proteins/*genetics ; Molecular Sequence Data ; Operon/genetics ; Phylogeny ; Polymerase Chain Reaction ; Sequence Alignment ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia is a genus of maternally inherited, intracellular, alpha-Proteobacteria which are widespread among arthropods and cause various reproductive alterations such as cytoplasmic incompatibility and feminization. Wolbachia strains in arthropods have been phylogenetically divided into two supergroups, A and B. Here, we determined the phylogenetic relationships among Wolbachia strains harbored by insects and isopod crustaceans by using groE operon sequences, which encode highly conserved bacterial heat shock proteins. This study includes the first survey for Wolbachia in isopods from a Southeast Asian country, i.e., Thailand. Two isopod species from different parts of Thailand were found to be infected by Wolbachia. Their groE sequences were also included in the phylogenetic analysis. Our results showed clearly that 19 strains from tropical insects, 11 strains from French isopods, and two strains from Thai isopods were in the B supergroup, and 15 strains from tropical insects were in the A supergroup. This is the first report of phylogenetic analysis of a large data set comprising Wolbachia groE sequences from both insects and isopod crustaceans.}, } @article {pmid19329661, year = {2009}, author = {Jin, C and Ren, X and Rasgon, JL}, title = {The virulent Wolbachia strain wMelPop efficiently establishes somatic infections in the malaria vector Anopheles gambiae.}, journal = {Applied and environmental microbiology}, volume = {75}, number = {10}, pages = {3373-3376}, pmid = {19329661}, issn = {1098-5336}, support = {R21 AI070178/AI/NIAID NIH HHS/United States ; R21 AI070178-02/AI/NIAID NIH HHS/United States ; R21AI070178/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology ; Microbial Viability ; Wolbachia/*growth & development/*isolation & purification ; }, abstract = {Wolbachia pipientis bacteria are maternally inherited endosymbionts that are of interest to control the Anopheles mosquito vectors of malaria. Wolbachia does not infect Anopheles mosquitoes in nature, although cultured Anopheles cells can be infected. Here, we show that the virulent Wolbachia strain wMelPop can survive and replicate when injected into female Anopheles gambiae adults, but the somatic infections established are avirulent. These in vivo data suggest that stable Wolbachia infections of Anopheles may be possible.}, } @article {pmid19317891, year = {2009}, author = {Charlat, S and Duplouy, A and Hornett, EA and Dyson, EA and Davies, N and Roderick, GK and Wedell, N and Hurst, GD}, title = {The joint evolutionary histories of Wolbachia and mitochondria in Hypolimnas bolina.}, journal = {BMC evolutionary biology}, volume = {9}, number = {}, pages = {64}, pmid = {19317891}, issn = {1471-2148}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Butterflies/*genetics/*microbiology ; DNA, Mitochondrial/genetics ; *Evolution, Molecular ; Genes, Insect ; Genes, Mitochondrial ; Haplotypes ; Host-Pathogen Interactions ; Inheritance Patterns ; Mitochondria/genetics ; Phylogeny ; Sequence Analysis, DNA ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: The interaction between the Blue Moon butterfly, Hypolimnas bolina, and Wolbachia has attracted interest because of the high prevalence of male-killing achieved within the species, the ecological consequences of this high prevalence, the intensity of selection on the host to suppress the infection, and the presence of multiple Wolbachia infections inducing different phenotypes. We examined diversity in the co-inherited marker, mtDNA, and the partitioning of this between individuals of different infection status, as a means to investigate the population biology and evolutionary history of the Wolbachia infections.

RESULTS: Part of the mitochondrial COI gene was sequenced from 298 individuals of known infection status revealing ten different haplotypes. Despite very strong biological evidence that the sample represents a single species, the ten haplotypes did not fall within a monophyletic clade within the Hypolimnas genus, with one haplotype differing by 5% from the other nine. There were strong associations between infection status and mtDNA haplotype. The presence of wBol1 infection in association with strongly divergent haplotypes prompted closer examination of wBol1 genetic variation. This revealed the existence of two cryptic subtypes, wBol1a and wBol1b. The wBol1a infection, by far the most common, was in strict association with the single divergent mtDNA haplotype. The wBol1b infection was found with two haplotypes that were also observed in uninfected specimens. Finally, the wBol2 infection was associated with a large diversity of mtDNA haplotypes, most often shared with uninfected sympatric butterflies.

CONCLUSION: This data overall supports the hypothesis that high prevalence of male-killing Wolbachia (wBol1) in H. bolina is associated with very high transmission efficiency rather than regular horizontal transmission. It also suggests this infection has undergone a recent selective sweep and was introduced in this species through introgression. In contrast, the sharing of haplotypes between wBol2-infected and uninfected individuals indicates that this strain is not perfectly transmitted and/or shows a significant level of horizontal transmission.}, } @article {pmid19307581, year = {2009}, author = {Klasson, L and Westberg, J and Sapountzis, P and Näslund, K and Lutnaes, Y and Darby, AC and Veneti, Z and Chen, L and Braig, HR and Garrett, R and Bourtzis, K and Andersson, SG}, title = {The mosaic genome structure of the Wolbachia wRi strain infecting Drosophila simulans.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {106}, number = {14}, pages = {5725-5730}, pmid = {19307581}, issn = {1091-6490}, mesh = {Animals ; Ankyrins/genetics ; Drosophila/*microbiology ; Gene Transfer, Horizontal ; Genes, Bacterial ; Genome, Bacterial/*genetics ; Molecular Sequence Data ; Mosaicism ; *Recombination, Genetic ; Wolbachia/*genetics ; }, abstract = {The obligate intracellular bacterium Wolbachia pipientis infects around 20% of all insect species. It is maternally inherited and induces reproductive alterations of insect populations by male killing, feminization, parthenogenesis, or cytoplasmic incompatibility. Here, we present the 1,445,873-bp genome of W. pipientis strain wRi that induces very strong cytoplasmic incompatibility in its natural host Drosophila simulans. A comparison with the previously sequenced genome of W. pipientis strain wMel from Drosophila melanogaster identified 35 breakpoints associated with mobile elements and repeated sequences that are stable in Drosophila lines transinfected with wRi. Additionally, 450 genes with orthologs in wRi and wMel were sequenced from the W. pipientis strain wUni, responsible for the induction of parthenogenesis in the parasitoid wasp Muscidifurax uniraptor. The comparison of these A-group Wolbachia strains uncovered the most highly recombining intracellular bacterial genomes known to date. This was manifested in a 500-fold variation in sequence divergences at synonymous sites, with different genes and gene segments supporting different strain relationships. The substitution-frequency profile resembled that of Neisseria meningitidis, which is characterized by rampant intraspecies recombination, rather than that of Rickettsia, where genes mostly diverge by nucleotide substitutions. The data further revealed diversification of ankyrin repeat genes by short tandem duplications and provided examples of horizontal gene transfer across A- and B-group strains that infect D. simulans. These results suggest that the transmission dynamics of Wolbachia and the opportunity for coinfections have created a freely recombining intracellular bacterial community with mosaic genomes.}, } @article {pmid19300496, year = {2009}, author = {Landmann, F and Orsi, GA and Loppin, B and Sullivan, W}, title = {Wolbachia-mediated cytoplasmic incompatibility is associated with impaired histone deposition in the male pronucleus.}, journal = {PLoS pathogens}, volume = {5}, number = {3}, pages = {e1000343}, pmid = {19300496}, issn = {1553-7374}, mesh = {Animals ; Animals, Genetically Modified ; Cell Nucleus ; Drosophila/genetics/*microbiology ; Embryo, Nonmammalian ; Female ; Fluorescent Antibody Technique ; Histones/*metabolism ; Host-Pathogen Interactions/*genetics ; Infertility, Male/genetics/microbiology ; Male ; Microscopy, Confocal ; Rickettsia Infections/*genetics ; Spermatozoa/pathology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a bacteria endosymbiont that rapidly infects insect populations through a mechanism known as cytoplasmic incompatibility (CI). In CI, crosses between Wolbachia-infected males and uninfected females produce severe cell cycle defects in the male pronucleus resulting in early embryonic lethality. In contrast, viable progeny are produced when both parents are infected (the Rescue cross). An important consequence of CI-Rescue is that infected females have a selective advantage over uninfected females facilitating the rapid spread of Wolbachia through insect populations. CI disrupts a number of prophase and metaphase events in the male pronucleus, including Cdk1 activation, chromosome condensation, and segregation. Here, we demonstrate that CI disrupts earlier interphase cell cycle events. Specifically, CI delays the H3.3 and H4 deposition that occurs immediately after protamine removal from the male pronucleus. In addition, we find prolonged retention of the replication factor PCNA in the male pronucleus into metaphase, indicating progression into mitosis with incompletely replicated DNA. We propose that these CI-induced interphase defects in de novo nucleosome assembly and replication are the cause of the observed mitotic condensation and segregation defects. In addition, these interphase chromosome defects likely activate S-phase checkpoints, accounting for the previously described delays in Cdk1 activation. These results have important implications for the mechanism of Rescue and other Wolbachia-induced phenotypes.}, } @article {pmid19296184, year = {2009}, author = {Fallon, AM and Witthuhn, BA}, title = {Proteasome activity in a naïve mosquito cell line infected with Wolbachia pipientis wAlbB.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {45}, number = {8}, pages = {460-466}, pmid = {19296184}, issn = {1543-706X}, support = {R21 AI070913/AI/NIAID NIH HHS/United States ; R21 AI070913-02/AI/NIAID NIH HHS/United States ; AI 070913/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/drug effects/*metabolism/*microbiology ; Amino Acid Sequence ; Animals ; Bromodeoxyuridine/pharmacology ; Cell Line ; Molecular Sequence Data ; Proteasome Endopeptidase Complex/*metabolism ; Ubiquitin/*metabolism ; Ubiquitination/physiology ; Up-Regulation/physiology ; Wolbachia/drug effects/*growth & development/*metabolism ; }, abstract = {We used Wolbachia pipientis strain wAlbB from Aedes albopictus Aa23 cells to infect clonal Ae. albopictus TK-6 cells, which are resistant to 5-bromodeoxyuridine. Infected TK-6 cells were cultured in medium containing 5-bromodeoxyuridine to select against Aa23 cells that might have persisted in the inoculum. Infected TK-6 lines retained the Wolbachia infection for 5 mo, indicating that their metabolic processes support Wolbachia growth and multiplication. To investigate early events after Wolbachia infection, we labeled infected cells with (35)S[methionine/cysteine]. Patterns of labeled proteins on sodium dodecyl sulfate gels were similar in control and infected cells, with the exception of a 29-kDa protein. Tandem mass spectrometry revealed that the 29-kDa band included alpha and beta subunits of the 26S proteasome. Independent confirmation of the up-regulation of the proteasome was established by probing Western blots with a monoclonal antibody to the proteasome-associated co-factor, ubiquitin. Wolbachia's loss of metabolic pathways for the synthesis of most amino acids and retention of pathways for their uptake and metabolism suggest that proteasome activation provides a mechanism whereby controlled degradation of intracellular host proteins would increase availability of amino acids to support establishment and maintenance of the Wolbachia infection.}, } @article {pmid19292821, year = {2009}, author = {Calvitti, M and Moretti, R and Porretta, D and Bellini, R and Urbanelli, S}, title = {Effects on male fitness of removing Wolbachia infections from the mosquito Aedes albopictus.}, journal = {Medical and veterinary entomology}, volume = {23}, number = {2}, pages = {132-140}, doi = {10.1111/j.1365-2915.2008.00791.x}, pmid = {19292821}, issn = {1365-2915}, mesh = {Aedes/*microbiology ; Animals ; Female ; Longevity ; Male ; Mosquito Control ; Pest Control, Biological ; Reproduction/physiology ; Selection, Genetic ; Wolbachia/*physiology ; }, abstract = {Cytoplasmic incompatibility (CI) induced by maternally inherited Wolbachia bacteria is a potential tool for the suppression of insect pest species with appropriate patterns of infection. The Asian tiger mosquito Aedes albopictus (Skuse) (Diptera: Culicidae) is known to be infected by two strains of Wolbachia pipientis Hertig (Rickettsiales: Rickettsiaceae), wAlb A and wAlb B, throughout its geographical distribution. This infection pattern theoretically restricts the application of CI-based control strategies. However, Wolbachia can be horizontally transferred using embryonic microinjection to generate incompatible transfected lines harbouring a single new strain of Wolbachia. In order to assess the feasibility of this approach, the effects of Wolbachia removal on mosquito fitness need to be clearly evaluated as the removal of natural superinfection is an inescapable step of this approach. Previous research has shown that uninfected females, produced by antibiotic treatment, showed a decrease in fitness compared with those infected with Wolbachia. In this study, the effect of Wolbachia removal on male fitness was investigated. Longevity and reproductive potential (mating competitiveness and sperm capacity) were assessed in both laboratory cages and greenhouses. No differences were observed between uninfected and infected males with respect to longevity, mating rate, sperm capacity and mating competitiveness in either laboratory conditions or greenhouses. The preservation of fitness in males of Ae. albopictus deprived of natural Wolbachia infection is discussed in relation to the development of incompatible insect technique suppression strategies. Finally, the potential application of aposymbiotic males in mark-release-recapture studies is suggested.}, } @article {pmid19284544, year = {2009}, author = {Nikoh, N and Nakabachi, A}, title = {Aphids acquired symbiotic genes via lateral gene transfer.}, journal = {BMC biology}, volume = {7}, number = {}, pages = {12}, pmid = {19284544}, issn = {1741-7007}, mesh = {Amino Acid Sequence ; Animal Structures/metabolism/microbiology ; Animals ; Aphids/cytology/*microbiology ; Buchnera/genetics/*physiology ; Carboxypeptidases/chemistry/genetics ; Gene Expression Profiling ; Gene Expression Regulation ; Gene Order ; Gene Transfer, Horizontal/*genetics ; Lipoprotein(a)/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Symbiosis/*genetics ; }, abstract = {BACKGROUND: Aphids possess bacteriocytes, which are cells specifically differentiated to harbour the obligate mutualist Buchnera aphidicola (gamma-Proteobacteria). Buchnera has lost many of the genes that appear to be essential for bacterial life. From the bacteriocyte of the pea aphid Acyrthosiphon pisum, we previously identified two clusters of expressed sequence tags that display similarity only to bacterial genes. Southern blot analysis demonstrated that they are encoded in the aphid genome. In this study, in order to assess the possibility of lateral gene transfer, we determined the full-length sequences of these transcripts, and performed detailed structural and phylogenetic analyses. We further examined their expression levels in the bacteriocyte using real-time quantitative RT-PCR.

RESULTS: Sequence similarity searches demonstrated that these fully sequenced transcripts are significantly similar to the bacterial genes ldcA (product, LD-carboxypeptidase) and rlpA (product, rare lipoprotein A), respectively. Buchnera lacks these genes, whereas many other bacteria, including Escherichia coli, a close relative of Buchnera, possess both ldcA and rlpA. Molecular phylogenetic analysis clearly demonstrated that the aphid ldcA was derived from a rickettsial bacterium closely related to the extant Wolbachia spp. (alpha-Proteobacteria, Rickettsiales), which are intracellular symbionts of various lineages of arthropods. The evolutionary origin of rlpA was not fully resolved, but it was clearly demonstrated that its double-psi beta-barrel domain is of bacterial origin. Real-time quantitative RT-PCR demonstrated that ldcA and rlpA are expressed 11.6 and 154-fold higher in the bacteriocyte than in the whole body, respectively. LdcA is an enzyme required for recycling murein (peptidoglycan), which is a component of the bacterial cell wall. As Buchnera possesses a cell wall composed of murein but lacks ldcA, a high level of expression of the aphid ldcA in the bacteriocyte may be essential to maintain Buchnera. Although the function of RlpA is not well known, conspicuous up-regulation of the aphid rlpA in the bacteriocyte implies that this gene is also essential for Buchnera.

CONCLUSION: In this study, we obtained several lines of evidence indicating that aphids acquired genes from bacteria via lateral gene transfer and that these genes are used to maintain the obligately mutualistic bacterium, Buchnera.}, } @article {pmid19281959, year = {2009}, author = {Merçot, H and Poinsot, D}, title = {Infection by Wolbachia: from passengers to residents.}, journal = {Comptes rendus biologies}, volume = {332}, number = {2-3}, pages = {284-297}, doi = {10.1016/j.crvi.2008.09.010}, pmid = {19281959}, issn = {1768-3238}, mesh = {Animals ; Arthropods/microbiology/physiology ; Bacterial Infections/*microbiology ; *Biological Evolution ; Reproduction/physiology ; Travel ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbiotic alpha-proteobacteria harboured by terrestrial arthropods and filarial nematodes, where they are maternally transmitted through egg cytoplasm. According to the host group, Wolbachia have developed two contrasting symbiotic strategies. In arthropods, symbiosis is secondary (i.e. facultative), and Wolbachia insure their transmission as reproduction parasites. However, despite of the efficiency of the manipulation mechanisms used, Wolbachia are limited to the state of passenger because some factors can prevent the association between Wolbachia and their hosts to become permanent. On the contrary, symbiosis is primary (i.e. obligatory) in filarial nematodes where Wolbachia insure their transmission via a mutualistic relationship, leading them to become permanent residents of their hosts. However, a few examples show that in arthropods too some Wolbachia have started to present the first stages of a mutualistic behaviour, or are even truly indispensable to their host. Whatever its strategy, Wolbachia infection is a spectacular evolutionary success, this symbiotic bacterium representing one of the most important biomass of its kind.}, } @article {pmid19271205, year = {2009}, author = {Mitreva, M and Smant, G and Helder, J}, title = {Role of horizontal gene transfer in the evolution of plant parasitism among nematodes.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {532}, number = {}, pages = {517-535}, doi = {10.1007/978-1-60327-853-9_30}, pmid = {19271205}, issn = {1064-3745}, support = {AI46593/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Cellulase/genetics ; Ecosystem ; Expressed Sequence Tags ; *Gene Transfer, Horizontal ; Genes, Helminth ; Host-Parasite Interactions/genetics ; Models, Genetic ; Nematoda/*genetics/*pathogenicity ; Phylogeny ; Plants/*parasitology ; Symbiosis/genetics ; }, abstract = {Horizontal gene transfer (HGT) implies the non-sexual exchange of genetic material between species - in some cases even across kingdoms. Although common among Bacteria and Archaea, HGTs from pro- to eukaryotes and between eukaryotes were thought to be extremely rare. Recent studies on intracellular bacteria and their hosts seriously question this view. Recipient organisms could benefit from HGT as new gene packages could allow them to broaden or change their diet, colonize new habitats, or survive conditions that previously would have been lethal.About a decade ago, plant parasitic nematodes were shown to produce and secrete cellulases. Prior to this, animals were thought to fully depend on microbial symbionts for the breakdown of plant cell walls. This discovery prompted Keen and Roberts (1) to hypothesize that the ability of nematodes to parasitize plants was acquired by HGT from soil bacteria to (ancestral) bacterivorous nematodes. Since the identification of the first nematode cellulases, many more plant cell wall-degrading enzymes (CWDE) have been identified in a range of plant parasitic nematode species.Here we discuss a number of criteria that can be used to underpin an HGT claim. HGT requires close physical contact between donor and recipient, and this could be achieved in, for example, a symbiont-host, or a trophic relationship. The former type of relationship was indeed shown to potentially result in the transfer of genetic material (e.g., Brugia malayi and Wolbachia). However, currently known endosymbionts of nematodes may not be the source of CWDEs. Remarkably, all cellulases discovered so far within the order Tylenchida belong to a single glycoside hydrolase family (GHF5). A range of soil bacteria harbours GHF5 cellulases, but of course nothing can be said about the gene content of soil bacteria at the time HGT took place (if at all). We suggest that characterisation of cellulases (and other CWDEs) and their genomic organisation in more basal (facultative) plant parasitic Tylenchida is needed to find out if CWDEs were indeed acquired via HGT from bacteria. A more complete picture about the evolution of CWDEs among plant parasitic Tylenchida will require a detailed characterisation of two - so far - fully unexplored basal suborders, Tylenchina and Criconematina. Finally, we performed a computational high-throughput identification of potential HGT candidates (including ones unrelated to CWDEs) in plant parasitic nematodes using a genomics approach.}, } @article {pmid19265485, year = {2009}, author = {Kurz, M and Iturbe-Ormaetxe, I and Jarrott, R and Shouldice, SR and Wouters, MA and Frei, P and Glockshuber, R and O'Neill, SL and Heras, B and Martin, JL}, title = {Structural and functional characterization of the oxidoreductase alpha-DsbA1 from Wolbachia pipientis.}, journal = {Antioxidants & redox signaling}, volume = {11}, number = {7}, pages = {1485-1500}, doi = {10.1089/ars.2008.2420}, pmid = {19265485}, issn = {1557-7716}, mesh = {Base Sequence ; Crystallography, X-Ray ; DNA Primers ; Oxidation-Reduction ; Oxidoreductases/chemistry/*metabolism ; Protein Conformation ; Protein Folding ; Structure-Activity Relationship ; Wolbachia/*enzymology ; }, abstract = {The alpha-proteobacterium Wolbachia pipientis is a highly successful intracellular endosymbiont of invertebrates that manipulates its host's reproductive biology to facilitate its own maternal transmission. The fastidious nature of Wolbachia and the lack of genetic transformation have hampered analysis of the molecular basis of these manipulations. Structure determination of key Wolbachia proteins will enable the development of inhibitors for chemical genetics studies. Wolbachia encodes a homologue (alpha-DsbA1) of the Escherichia coli dithiol oxidase enzyme EcDsbA, essential for the oxidative folding of many exported proteins. We found that the active-site cysteine pair of Wolbachia alpha-DsbA1 has the most reducing redox potential of any characterized DsbA. In addition, Wolbachia alpha-DsbA1 possesses a second disulfide that is highly conserved in alpha-proteobacterial DsbAs but not in other DsbAs. The alpha-DsbA1 structure lacks the characteristic hydrophobic features of EcDsbA, and the protein neither complements EcDsbA deletion mutants in E. coli nor interacts with EcDsbB, the redox partner of EcDsbA. The surface characteristics and redox profile of alpha-DsbA1 indicate that it probably plays a specialized oxidative folding role with a narrow substrate specificity. This first report of a Wolbachia protein structure provides the basis for future chemical genetics studies.}, } @article {pmid19257899, year = {2009}, author = {Delgado, AM and Cook, JM}, title = {Effects of a sex-ratio distorting endosymbiont on mtDNA variation in a global insect pest.}, journal = {BMC evolutionary biology}, volume = {9}, number = {}, pages = {49}, pmid = {19257899}, issn = {1471-2148}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; DNA, Bacterial/genetics ; DNA, Mitochondrial/*genetics ; Evolution, Molecular ; Female ; Genes, Insect ; Genetic Variation ; *Genetics, Population ; Geography ; Haplotypes ; Male ; Mitochondria/genetics ; Moths/*genetics/microbiology ; Phenotype ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; *Sex Ratio ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Patterns of mtDNA variation within a species reflect long-term population structure, but may also be influenced by maternally inherited endosymbionts, such as Wolbachia. These bacteria often alter host reproductive biology and can drive particular mtDNA haplotypes through populations. We investigated the impacts of Wolbachia infection and geography on mtDNA variation in the diamondback moth, a major global pest whose geographic distribution reflects both natural processes and transport via human agricultural activities.

RESULTS: The mtDNA phylogeny of 95 individuals sampled from 10 countries on four continents revealed two major clades. One contained only Wolbachia-infected individuals from Malaysia and Kenya, while the other contained only uninfected individuals, from all countries including Malaysia and Kenya. Within the uninfected group was a further clade containing all individuals from Australasia and displaying very limited sequence variation. In contrast, a biparental nuclear gene phylogeny did not have infected and uninfected clades, supporting the notion that maternally-inherited Wolbachia are responsible for the mtDNA pattern. Only about 5% (15/306) of our global sample of individuals was infected with the plutWB1 isolate and even within infected local populations, many insects were uninfected. Comparisons of infected and uninfected isofemale lines revealed that plutWB1 is associated with sex ratio distortion. Uninfected lines have a 1:1 sex ratio, while infected ones show a 2:1 female bias.

CONCLUSION: The main correlate of mtDNA variation in P. xylostella is presence or absence of the plutWB1 infection. This is associated with substantial sex ratio distortion and the underlying mechanisms deserve further study. In contrast, geographic origin is a poor predictor of moth mtDNA sequences, reflecting human activity in moving the insects around the globe. The exception is a clade of Australasian individuals, which may reflect a bottleneck during their recent introduction to this region.}, } @article {pmid19253762, year = {2009}, author = {Nogrady, B}, title = {Bug for a bug for a bug.}, journal = {Scientific American}, volume = {300}, number = {3}, pages = {17}, doi = {10.1038/scientificamerican0309-17}, pmid = {19253762}, issn = {0036-8733}, mesh = {Aedes/*microbiology/physiology ; Animals ; Dengue/prevention & control ; Female ; Insect Bites and Stings/prevention & control ; Male ; Mosquito Control/*methods ; Pest Control, Biological ; Wolbachia/*physiology ; }, } @article {pmid19229488, year = {2009}, author = {Jeong, G and Lee, K and Choi, J and Hwang, S and Park, B and Kim, W and Choi, Y and Park, I and Kim, J}, title = {Incidence of Wolbachia and Cardinium endosymbionts in the Osmia community in Korea.}, journal = {Journal of microbiology (Seoul, Korea)}, volume = {47}, number = {1}, pages = {28-32}, pmid = {19229488}, issn = {1976-3794}, mesh = {Animals ; Bacteroidetes/genetics/*isolation & purification ; Bees/genetics/*microbiology/parasitology ; Diptera/genetics/microbiology ; Gene Transfer, Horizontal ; Korea ; Mites/genetics/microbiology ; Pest Control, Biological ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Sex Ratio ; *Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Sex ratio distorting endosymbionts induce reproductive anomalies in their arthropod hosts. They have recently been paid much attention as firstly texts of evolution of host-symbiont relationships and secondly potential biological control agents to control arthropod pests. Among such organisms, Wolbachia and Cardinium bacteria are well characterized. This study aims at probing such bacteria in the Osmia community to evaluate their potential utilization to control arthropod pests. Among 17 PCR tested species, Osmia cornifrons and a parasitic fly are infected with Wolbachia and a mite species is infected with Cardinium. Phylogenetic tree analyses suggest that horizontal transfer of the bacteria occurred between phylogenetically distant hosts.}, } @article {pmid19226322, year = {2009}, author = {Haselkorn, TS and Markow, TA and Moran, NA}, title = {Multiple introductions of the Spiroplasma bacterial endosymbiont into Drosophila.}, journal = {Molecular ecology}, volume = {18}, number = {6}, pages = {1294-1305}, doi = {10.1111/j.1365-294X.2009.04085.x}, pmid = {19226322}, issn = {1365-294X}, mesh = {Animals ; Bacterial Typing Techniques ; Bayes Theorem ; *Biological Evolution ; DNA, Bacterial/genetics ; DNA, Mitochondrial/genetics ; Drosophila/genetics/*microbiology ; Genetic Variation ; Genetics, Population ; Haplotypes ; Mitochondria/genetics ; *Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Spiroplasma/classification/*genetics ; *Symbiosis ; }, abstract = {Bacterial endosymbionts are common in insects and can have dramatic effects on their host's evolution. So far, the only heritable symbionts found in Drosophila have been Wolbachia and Spiroplasma. While the incidence and effects of Wolbachia have been studied extensively, the prevalence and significance of Spiroplasma infections in Drosophila are less clear. These small, gram-positive, helical bacteria infect a diverse array of plant and arthropod hosts, conferring a variety of fitness effects. Male-killing Spiroplasma are known from certain Drosophila species; however, in others, Spiroplasma appear not to affect sex ratio. Previous studies have identified different Spiroplasma haplotypes in Drosophila populations, although no extensive surveys have yet been reported. We used a multilocus sequence analysis to reconstruct a robust Spiroplasma endosymbiont phylogeny, assess genetic diversity, and look for evidence of recombination. Six loci were sequenced from over 65 Spiroplasma-infected individuals from nine different Drosophila species. Analysis of these sequences reveals at least five separate introductions of four phylogenetically distinct Spiroplasma haplotypes, indicating that more extensive sampling will likely reveal an even greater Spiroplasma endosymbiont diversity. Patterns of variation in Drosophila mitochondrial haplotypes in Spiroplasma-infected and uninfected flies imply imperfect vertical transmission in host populations and possible horizontal transmission.}, } @article {pmid19223923, year = {2009}, author = {Ros, VI and Breeuwer, JA}, title = {The effects of, and interactions between, Cardinium and Wolbachia in the doubly infected spider mite Bryobia sarothamni.}, journal = {Heredity}, volume = {102}, number = {4}, pages = {413-422}, doi = {10.1038/hdy.2009.4}, pmid = {19223923}, issn = {1365-2540}, mesh = {Animals ; Bacteroidetes/pathogenicity/*physiology ; Cytoplasm/metabolism/microbiology ; Female ; Male ; Symbiosis ; Tetranychidae/genetics/*microbiology/parasitology ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Many arthropods are infected with vertically transmitted, intracellular bacteria manipulating their host's reproduction. Cytoplasmic incompatibility (CI) is commonly observed and is expressed as a reduction in the number of offspring in crosses between infected males and uninfected females (or females infected with a different bacterial strain). CI is often related to the presence of Wolbachia, but recent findings indicate that a second reproductive parasite, Cardinium, is also capable of inducing CI. Although both Wolbachia and Cardinium occur in arthropods and may infect the same host species, little is known about their interactions. We observed Wolbachia and Cardinium in the sexual spider mite Bryobia sarothamni (Acari: Tetranychidae) and investigated the effects of both bacteria on reproduction. We performed all possible crossing combinations using naturally infected strains, and show that Cardinium induces strong CI, expressed as an almost complete female mortality. B. sarothamni is the third host species in which Cardinium-induced CI is observed, and this study reveals the strongest CI effect found so far. Wolbachia, however, did not induce CI. Even so, CI was not induced by doubly infected males, and neither singly Wolbachia-infected nor doubly infected females could rescue CI induced by Cardinium-infected males. Possibly, this is related to the differences between Cardinium strains infecting singly and doubly infected individuals. We found a cost of infection in single infected individuals, but not in doubly infected individuals. We show that infection frequencies in field populations ranged from completely uninfected to a polymorphic state. In none of the populations infections were fixed.}, } @article {pmid19223921, year = {2009}, author = {White, JA and Kelly, SE and Perlman, SJ and Hunter, MS}, title = {Cytoplasmic incompatibility in the parasitic wasp Encarsia inaron: disentangling the roles of Cardinium and Wolbachia symbionts.}, journal = {Heredity}, volume = {102}, number = {5}, pages = {483-489}, pmid = {19223921}, issn = {1365-2540}, support = {K12 GM000708/GM/NIGMS NIH HHS/United States ; K12 GM00708-06/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteroidetes/*physiology ; Extrachromosomal Inheritance ; Female ; *Host-Pathogen Interactions ; Male ; Reproduction ; Sex Ratio ; *Symbiosis ; Wasps/*genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Many bacterial endosymbionts of insects are capable of manipulating their host's reproduction for their own benefit. The most common strategy of manipulation is cytoplasmic incompatibility (CI), in which embryonic mortality results from matings between uninfected females and infected males. In contrast, embryos develop normally in infected females, whether or not their mate is infected, and infected progeny are produced. In this way, the proportion of infected females increases in the insect population, thereby promoting the spread of the maternally inherited bacteria. However, what happens when multiple endosymbionts inhabit the same host? The parasitoid wasp Encarsia inaron is naturally infected with two unrelated endosymbionts, Cardinium and Wolbachia, both of which have been documented to cause CI in other insects. Doubly infected wasps show the CI phenotype. We differentially cured E. inaron of each endosymbiont, and crossed hosts of different infection status to determine whether either or both bacteria caused the observed CI phenotype in this parasitoid, and whether the two symbionts interacted within their common host. We found that Wolbachia caused CI in E. inaron, but Cardinium did not. We did not find evidence that Cardinium was able to modify or rescue Wolbachia-induced CI, nor did we find that Cardinium caused progeny sex ratio distortion, leaving the role of Cardinium in E. inaron a mystery.}, } @article {pmid19222304, year = {2008}, author = {Teixeira, L and Ferreira, A and Ashburner, M}, title = {The bacterial symbiont Wolbachia induces resistance to RNA viral infections in Drosophila melanogaster.}, journal = {PLoS biology}, volume = {6}, number = {12}, pages = {e2}, pmid = {19222304}, issn = {1545-7885}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Drosophila melanogaster/drug effects/immunology/*microbiology/*virology ; Female ; Immunity, Innate/drug effects ; Male ; RNA Viruses/drug effects/*physiology ; Reproducibility of Results ; *Symbiosis/drug effects ; Tetracycline/pharmacology ; Virus Diseases/*immunology/*microbiology ; Wolbachia/drug effects/*physiology ; }, abstract = {Wolbachia are vertically transmitted, obligatory intracellular bacteria that infect a great number of species of arthropods and nematodes. In insects, they are mainly known for disrupting the reproductive biology of their hosts in order to increase their transmission through the female germline. In Drosophila melanogaster, however, a strong and consistent effect of Wolbachia infection has not been found. Here we report that a bacterial infection renders D. melanogaster more resistant to Drosophila C virus, reducing the load of viruses in infected flies. We identify these resistance-inducing bacteria as Wolbachia. Furthermore, we show that Wolbachia also increases resistance of Drosophila to two other RNA virus infections (Nora virus and Flock House virus) but not to a DNA virus infection (Insect Iridescent Virus 6). These results identify a new major factor regulating D. melanogaster resistance to infection by RNA viruses and contribute to the idea that the response of a host to a particular pathogen also depends on its interactions with other microorganisms. This is also, to our knowledge, the first report of a strong beneficial effect of Wolbachia infection in D. melanogaster. The induced resistance to natural viral pathogens may explain Wolbachia prevalence in natural populations and represents a novel Wolbachia-host interaction.}, } @article {pmid19209229, year = {2009}, author = {Koehncke, A and Telschow, A and Werren, JH and Hammerstein, P}, title = {Life and death of an influential passenger: Wolbachia and the evolution of CI-modifiers by their hosts.}, journal = {PloS one}, volume = {4}, number = {2}, pages = {e4425}, pmid = {19209229}, issn = {1932-6203}, mesh = {Animals ; *Biological Evolution ; Cytoplasm/*metabolism ; Female ; *Host-Pathogen Interactions ; Male ; Microbial Viability ; Mutation/genetics ; Sex Characteristics ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia are intracellular bacteria widely distributed among arthropods and nematodes. In many insect species these bacteria induce a cytoplasmic incompatibility (CI) between sperm of infected males and eggs of uninfected females. From an evolutionary point of view, CI is puzzling: In order to induce this modification-rescue system, Wolbachia affect sperm of infected males even though Wolbachia are only transmitted maternally. Phylogenetic studies of Wolbachia and hosts show that the bacteria rarely cospeciate with their hosts, indicating that infections are lost in host species. However, the mechanisms leading to Wolbachia loss are not well understood.

RESULTS: Using a population genetic model, we investigate the spread of host mutants that enhance or repress Wolbachia action by affecting either bacterial transmission or the level of CI. We show that host mutants that decrease CI-levels in males (e.g. by reducing Wolbachia-density during spermatogenesis) spread, even at cost to mutant males. Increase of these mutants can lead to loss of Wolbachia infections, either as a direct consequence of their increase or in a step-wise manner, and we derive analytically a threshold penetrance above which a mutation's spread leads to extinction of Wolbachia. Selection on host modifiers is sexually antagonistic in that, conversely, host mutants that enhance Wolbachia in females are favoured whereas suppressors are not.

CONCLUSIONS: Our results indicate that Wolbachia is likely to be lost from host populations on long evolutionary time scales due to reduction of CI levels in males. This can occur either by evolution of single host modifiers with large effects or through accumulation of several modifier alleles with small effects on Wolbachia action, even at cost to mutant males and even if infected hosts do not incur fecundity costs. This possibility is consistent with recent findings and may help to explain the apparent short evolutionary persistence times of Wolbachia in many host systems.}, } @article {pmid19198544, year = {2008}, author = {Zhukova, MV and Voronin, DA and Kiseleva, EV}, title = {[High temperature initiates changes of Wolbachia ultrastructure in the ovaries and early embryos of Drosophila melanogaster].}, journal = {Tsitologiia}, volume = {50}, number = {12}, pages = {1050-1060}, pmid = {19198544}, issn = {0041-3771}, mesh = {Animals ; Drosophila melanogaster/embryology/*microbiology ; Embryo, Nonmammalian/*microbiology/ultrastructure ; Female ; Genome, Insect ; Hot Temperature ; Ovary/*microbiology/ultrastructure ; Symbiosis ; Wolbachia/physiology/*ultrastructure ; }, abstract = {Electron microscopic analysis of Drosophila melanogaster (w1118) ovarian cells has demonstrated that stressful heat treatment of flies results in the appearance of electron dense granules and large lysosomes in the cytoplasm of ovarian cells, which is not related with the presence of Wolbachia, as these changes are observed in both the infected and uninfected flies. High temperature initiates essential envelope defects and other structural changes of symbiotic bacteria in the cytoplasm of ovarian cells. Some embryos developing from eggs of heat shocked flies die, however, bacteria in the survival embryos retain their typical morphology. Endosymbionts do not change their localization and their contacts with the mitochondria and endoplasmic reticulum in the ovarian cells and early embryos after heat shock treatment of the flies. The results obtained show that high temperature influences on both the host and the endosymbiont, but does not change their structural mutual interactions.}, } @article {pmid19181828, year = {2009}, author = {Mouton, L and Henri, H and Fleury, F}, title = {Interactions between coexisting intracellular genomes: mitochondrial density and Wolbachia infection.}, journal = {Applied and environmental microbiology}, volume = {75}, number = {7}, pages = {1916-1921}, pmid = {19181828}, issn = {1098-5336}, mesh = {Animals ; Drosophila/*genetics/*microbiology ; *Host-Parasite Interactions ; Mitochondria/*genetics ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Many arthropods are infected with maternally transmitted microorganisms, leading to the coexistence of several intracellular genomes within the host cells, including their own mitochondria. As these genomes are cotransmitted, their patterns of evolution have been intimately linked, with possible consequences for the diversity and evolution of the host mitochondrial DNA. The evolutionary aspects of the situation have been thoroughly investigated, especially the selective sweep on the mitochondria as a result of Wolbachia invasion, whereas direct interactions between mitochondria and intracellular symbionts within the host cells or body have received little attention. Since endosymbionts exploit host resources but mitochondria supply energy to meet the bioenergetic demands of organisms, an unanswered question concerns the correlation between their densities. Here, we investigated the influence of Wolbachia symbiosis on mitochondrial density in two parasitic wasps of Drosophila species, both of which are naturally infected by three Wolbachia strains, but they differ in their degree of dependency on these bacteria. In Leptopilina heterotoma, all Wolbachia strains are facultative, whereas Asobara tabida requires a strain of Wolbachia for oogenesis to occur. In both species, Wolbachia infections are stable and well regulated, since the density of each strain does not depend on the presence or absence of other strains. Using lines that harbor various Wolbachia infection statuses, we found that mitochondrial density was not affected by the infection regardless of the sex and age of the host, which is strongly reminiscent of the independent regulation of specific Wolbachia strains and suggest that the protagonists coexist independently of each other as the result of a long-term coevolutionary interaction.}, } @article {pmid19171941, year = {2009}, author = {Sun, S and Cline, TW}, title = {Effects of Wolbachia infection and ovarian tumor mutations on Sex-lethal germline functioning in Drosophila.}, journal = {Genetics}, volume = {181}, number = {4}, pages = {1291-1301}, pmid = {19171941}, issn = {0016-6731}, support = {R01 GM023468/GM/NIGMS NIH HHS/United States ; GM23468/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila Proteins/*genetics/metabolism/*physiology ; Drosophila melanogaster/*genetics/physiology ; Epistasis, Genetic/physiology ; Female ; Gene Dosage ; Gene Expression Regulation, Developmental ; Germ Cells/*metabolism/physiology ; Mutation/physiology ; Ovary/growth & development/metabolism ; RNA-Binding Proteins/genetics/metabolism/*physiology ; Recombination, Genetic ; Rickettsiaceae Infections/*genetics/physiopathology/veterinary ; Vitellogenesis/genetics ; Vitellogenins/genetics/metabolism ; *Wolbachia/physiology ; X Chromosome/genetics ; }, abstract = {Wolbachia is a ubiquitous intracellular endosymbiont of invertebrates. Surprisingly, infection of Drosophila melanogaster by this maternally inherited bacterium restores fertility to females carrying ovarian tumor (cystocyte overproliferation) mutant alleles of the Drosophila master sex-determination gene, Sex-lethal (Sxl). We scanned the Drosophila genome for effects of infection on transcript levels in wild-type previtellogenic ovaries that might be relevant to this suppression of female-sterile Sxl mutants by Wolbachia. Yolk protein gene transcript levels were most affected, being reduced by infection, but no genes showed significantly more than a twofold difference. The yolk gene effect likely signals a small, infection-induced delay in egg chamber maturation unrelated to suppression. In a genetic study of the Wolbachia-Sxl interaction, we established that germline Sxl controls meiotic recombination as well as cystocyte proliferation, but Wolbachia only influences the cystocyte function. In contrast, we found that mutations in ovarian tumor (otu) interfere with both Sxl germline functions. We were led to otu through characterization of a spontaneous dominant suppressor of the Wolbachia-Sxl interaction, which proved to be an otu mutation. Clearly Sxl and otu work together in the female germline. These studies of meiosis in Sxl mutant females revealed that X chromosome recombination is considerably more sensitive than autosomal recombination to reduced Sxl activity.}, } @article {pmid19169549, year = {2008}, author = {Copeland, CS and Matthews, RW and González, JM and Aluja, M and Sivinski, J}, title = {Wolbachia in two populations of Melittobia digitata Dahms (Hymenoptera: Eulophidae).}, journal = {Neotropical entomology}, volume = {37}, number = {6}, pages = {633-640}, doi = {10.1590/s1519-566x2008000600002}, pmid = {19169549}, issn = {1519-566X}, mesh = {Animals ; Female ; Hymenoptera/*microbiology ; Male ; Polymerase Chain Reaction ; Wolbachia/*isolation & purification ; }, abstract = {We investigated two populations of Melittobia digitata Dahms, a gregarious parasitoid (primarily upon a wide range of solitary bees, wasps, and flies), in search of Wolbachia infection. The first population, from Xalapa, Mexico, was originally collected from and reared on Mexican fruit fly pupae, Anastrepha ludens Loew (Diptera: Tephritidae); the other, from Athens, Georgia, was collected from and reared on prepupae of mud dauber wasps, Trypoxylon politum Say (Hymenoptera: Crabronidae). PCR studies of the ITS2 region corroborated that both parasitoid populations were the same species; this potentially provides a useful molecular taxonomic profile since females of Melittobia species are superficially similar. Amplification of the Wolbachia surface protein gene (wsp) confirmed the presence of this endosymbiont in both populations. Sequencing revealed that the Wolbachia harbored in both populations exhibited a wsp belonging to a unique subgroup (denoted here as Dig) within the B-supergroup of known wsp genes. This new subgroup of wsp may either belong to a different strain of Wolbachia from those previously found to infect Melittobia or may be the result of a recombination event. In either case, known hosts of Wolbachia with a wsp of this subgroup are only distantly related taxonomically. Reasons are advanced as to why Melittobia - an easily reared and managed parasitoid - holds promise as an instructive model organism of Wolbachia infection amenable to the investigation of Wolbachia strains among its diverse hosts.}, } @article {pmid19168746, year = {2009}, author = {Gentil, K and Pearlman, E}, title = {Gamma interferon and interleukin-1 receptor 1 regulate neutrophil recruitment to the corneal stroma in a murine model of Onchocerca volvulus keratitis.}, journal = {Infection and immunity}, volume = {77}, number = {4}, pages = {1606-1612}, pmid = {19168746}, issn = {1098-5522}, support = {P30 EY011373/EY/NEI NIH HHS/United States ; R01 EY010320/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Cell Line ; Cornea/cytology/parasitology ; Corneal Stroma/*immunology/parasitology ; Fibroblasts/parasitology ; Humans ; Interferon-gamma/genetics/*metabolism ; Keratitis/*immunology/parasitology ; Macrophages/immunology/parasitology ; Mice ; Mice, Inbred C57BL ; Neutrophil Infiltration/*immunology ; Onchocerca volvulus/*pathogenicity ; Onchocerciasis/*immunology/parasitology ; Receptors, Interleukin-1 Type I/genetics/*metabolism ; Toll-Like Receptor 2/metabolism ; }, abstract = {Toll-like receptor 2 (TLR2) is an essential mediator of corneal inflammation induced by the filarial nematode Onchocerca volvulus, which harbors endosymbiotic Wolbachia bacteria. TLR2 is also required for dendritic cell activation, gamma interferon (IFN-gamma) production, and neutrophil recruitment to the cornea. To examine the role of IFN-gamma in O. volvulus keratitis, C57BL/6 and IFN-gamma(-/-) mice were immunized subcutaneously, and a soluble antigen extract from O. volvulus adult worms (OvAg) was injected into the corneal stroma of each animal. We found that, in the absence of IFN-gamma, neutrophil recruitment to the cornea was significantly impaired, whereas there was no effect on eosinophil infiltration. Since the cornea contains resident macrophages and fibroblasts and our previous studies showed that CXC chemokines mediate neutrophil recruitment, we examined the role of recombinant IFN-gamma (rIFN-gamma) on each cell type. We found no effect of rIFN-gamma on CXC chemokine production by macrophages or corneal fibroblasts, either alone or with filarial extracts; in contrast, rIFN-gamma was found to enhance OvAg-induced tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), IL-1alpha, and IL-1beta in macrophages. Furthermore, we found that rTNF-alpha, rIL-1alpha, or rIL-1beta induced CXC chemokine production by corneal fibroblasts but not by macrophages. To determine the relative contributions of endogenous cytokines, we injected OvAg into the corneal stroma of C57BL/6, IL-1 receptor 1(-/-) (IL-1R1(-/-)), and TNF-alphaR1/2(-/-) mice and examined neutrophil recruitment. We found that neutrophil infiltration was impaired in IL-1R1(-/-) mice but not in TNF-alphaR1/2(-/-) mice. IFN-gamma therefore appears to regulate neutrophil recruitment to the corneal stroma by enhancing TLR2 expression and OvAg-induced IL-1alpha and IL-1beta production by macrophages in the cornea, which then induce IL-1R1-dependent production of CXC chemokine by resident corneal fibroblasts.}, } @article {pmid19165180, year = {2009}, author = {Harrington, M}, title = {Bacterial infection may limit spread of dengue fever.}, journal = {Lab animal}, volume = {38}, number = {2}, pages = {40}, doi = {10.1038/laban0209-40b}, pmid = {19165180}, issn = {0093-7355}, mesh = {Aedes/*virology ; Animals ; Dengue/microbiology/*prevention & control ; *Gram-Negative Bacterial Infections ; Host-Pathogen Interactions ; Insect Vectors/virology ; Mosquito Control/*methods ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, } @article {pmid19161685, year = {2008}, author = {Kumm, S and Moritz, G}, title = {First detection of Wolbachia in arrhenotokous populations of thrips species (Thysanoptera: Thripidae and Phlaeothripidae) and its role in reproduction.}, journal = {Environmental entomology}, volume = {37}, number = {6}, pages = {1422-1428}, doi = {10.1603/0046-225x-37.6.1422}, pmid = {19161685}, issn = {0046-225X}, mesh = {Animals ; Anti-Bacterial Agents ; DNA, Bacterial/analysis ; Female ; Hot Temperature ; Insecta/*microbiology/physiology ; Male ; *Parthenogenesis ; Sequence Analysis, DNA ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Insects in the order Thysanoptera are known for their haplodiploid reproduction, with most species reproducing by arrhenotoky. A few species, however, have only female populations and reproduce by thelytoky. Seven thrips species reproducing either by arrhenotoky [Frankliniella occidentalis (Pergande), Echinothrips americanus Morgan, Suocerathrips linguis Mound and Marullo, Gynaikothrips ficorum (Marchal)] or thelytoky [Hercinothrips femoralis (O.M. Reuter) and Parthenothrips dracaenae (Heeger)] or showing both reproduction modes (Thrips tabaci Lindeman) were screened for the presence of Wolbachia using 16S rDNA and ftsZ gene primers. The arrhenotokous reproducing species E. americanus, G. ficorum, and S. linguis tested positive for Wolbachia. The bacterium was not detected in F. occidentalis or T. tabaci; even the thelytokous population of T. tabaci was free of Wolbachia. Wolbachia was found in the thelytokous reproducing species H. femoralis and P. dracaenae. Antibiotic treatment of H. femoralis induced the production of males that copulated with females. The results confirmed Wolbachia to be present in thelytokous reproducing thrips species and proved the presence of the bacterium for the first time in some arrhenotokous reproducing thrips species.}, } @article {pmid19154594, year = {2009}, author = {Klasson, L and Kambris, Z and Cook, PE and Walker, T and Sinkins, SP}, title = {Horizontal gene transfer between Wolbachia and the mosquito Aedes aegypti.}, journal = {BMC genomics}, volume = {10}, number = {}, pages = {33}, pmid = {19154594}, issn = {1471-2164}, support = {/WT_/Wellcome Trust/United Kingdom ; 079059/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*genetics/*microbiology ; Animals ; DNA, Bacterial/genetics ; *Evolution, Molecular ; Female ; *Gene Transfer, Horizontal ; Genes, Insect ; Genome, Bacterial ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: The evolutionary importance of horizontal gene transfer (HGT) from Wolbachia endosymbiotic bacteria to their eukaryotic hosts is a topic of considerable interest and debate. Recent transfers of genome fragments from Wolbachia into insect chromosomes have been reported, but it has been argued that these fragments may be on an evolutionary trajectory to degradation and loss.

RESULTS: We have discovered a case of HGT, involving two adjacent genes, between the genomes of Wolbachia and the currently Wolbachia-uninfected mosquito Aedes aegypti, an important human disease vector. The lower level of sequence identity between Wolbachia and insect, the transcription of all the genes involved, and the fact that we have identified homologs of the two genes in another Aedes species (Ae. mascarensis), suggest that these genes are being expressed after an extended evolutionary period since horizontal transfer, and therefore that the transfer has functional significance. The association of these genes with Wolbachia prophage regions also provides a mechanism for the transfer.

CONCLUSION: The data support the argument that HGT between Wolbachia endosymbiotic bacteria and their hosts has produced evolutionary innovation.}, } @article {pmid19145467, year = {2009}, author = {De Luna, CJ and Moro, CV and Guy, JH and Zenner, L and Sparagano, OA}, title = {Endosymbiotic bacteria living inside the poultry red mite (Dermanyssus gallinae).}, journal = {Experimental & applied acarology}, volume = {48}, number = {1-2}, pages = {105-113}, pmid = {19145467}, issn = {1572-9702}, mesh = {Animals ; Bacteroidetes/classification/genetics/*isolation & purification ; Base Sequence ; Coxiellaceae/classification/genetics/*isolation & purification ; France ; Mites/*microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Spiroplasma/classification/genetics/*isolation & purification ; Symbiosis ; United Kingdom ; Xanthomonadaceae/classification/genetics/*isolation & purification ; }, abstract = {This study investigated the endosymbiotic bacteria living inside the poultry red mite collected from five samples of one commercial farm from the UK and 16 farms from France using genus-specific PCR, PCR-TTGE and DNA sequencing. Endosymbiotic bacteria are intracellular obligate organisms that can cause several phenotypic and reproductive anomalies to their host and they are found widespread living inside arthropods. The farm sampled from the UK was positive for bacteria of the genera Cardinium sp. and Spiroplasma sp. From France, 7 farms were positive for Cardinium sp., 1 farm was positive for Spiroplasma sp., 1 farm was positive for Rickettsiella sp. and 2 farms were positive for Schineria sp. However, it was not possible to detect the presence of the genus Wolbachia sp. which has been observed in other ectoparasites. This study is the first report of the presence of endosymbionts living inside the poultry red mite. The results obtained suggest that it may be possible that these bacterial endosymbionts cause biological modifications to the poultry red mite.}, } @article {pmid19142204, year = {2009}, author = {Sheeley, SL and McAllister, BF}, title = {Mobile male-killer: similar Wolbachia strains kill males of divergent Drosophila hosts.}, journal = {Heredity}, volume = {102}, number = {3}, pages = {286-292}, doi = {10.1038/hdy.2008.126}, pmid = {19142204}, issn = {1365-2540}, mesh = {Animals ; Drosophila/genetics/*microbiology ; Female ; *Host-Pathogen Interactions ; Male ; Ovary/microbiology ; Phenotype ; Phylogeny ; Sex Ratio ; Species Specificity ; Wolbachia/classification/genetics/isolation & purification/*physiology ; Y Chromosome/genetics ; }, abstract = {Wolbachia are capable of eliciting a variety of reproductive phenotypes from their hosts, including the production of an all-female progeny through embryonic male-killing. To date, phylogenetic analyses indicate six independent acquisitions of the ability to kill male embryos among Wolbachia strains which infect insects. Of these six strains, only one appears to have experienced horizontal transmission between host species while maintaining a male-killing phenotype. The rarity of male-killing Wolbachia and their disjunct phylogenetic relationships is surprising, given the apparently common occurrence of horizontal transfer involving Wolbachia strains causing other phenotypes. A male-killing Wolbachia strain examined here in Drosophila borealis represents a second case of apparent horizontal transmission, based on its close relationship to a male-killing strain in a distantly related Drosophila species. The results reported here show that this Wolbachia has maintained a stable phenotype in D. borealis over a period of at least 50 years, and that a similar strain elicits the same male-killing phenotype in a second Drosophila species, indicating that male-killing may be a stable long-term strategy. Sampling bias and/or a lack of suitable hosts are discussed as possible causes of the low frequency of male-killers identified among Wolbachia strains.}, } @article {pmid19133077, year = {2008}, author = {Tortosa, P and Courtiol, A and Moutailler, S and Failloux, AB and Weill, M}, title = {Chikungunya-Wolbachia interplay in Aedes albopictus.}, journal = {Insect molecular biology}, volume = {17}, number = {6}, pages = {677-684}, doi = {10.1111/j.1365-2583.2008.00842.x}, pmid = {19133077}, issn = {1365-2583}, mesh = {Aedes/*microbiology/*virology ; Animals ; Bacteriophages/*genetics ; Chikungunya virus/*genetics ; DNA Primers/genetics ; Female ; Host-Parasite Interactions ; *Insect Vectors ; Plasmids/genetics ; Polymerase Chain Reaction ; Reunion ; Sex Factors ; Virus Replication/physiology ; Wolbachia/genetics/*virology ; }, abstract = {A severe Chikungunya (CHIK) outbreak recently hit several countries of the Indian Ocean. On La Réunion Island, Aedes albopictus was incriminated as the major vector. This mosquito species is naturally co-infected with two distinct strains of the endosymbiont Wolbachia, namely wAlbA and wAlbB, which are increasingly attracting interest as potential tools for vector control. A PCR quantitative assay was developed to investigate Wolbachia/mosquito host interactions. We show that Wolbachia densities are slightly decreased in CHIK virus (CHIKV)-infected females. We measured the impact of CHIKV replication on a lysogenic virus: WO bacteriophage. Our data indicate that WO is sheltered by wAlbB, likely at a single copy per bacteria, and that CHIKV replication is not a physiological stress triggering WO entrance into the lytic cycle.}, } @article {pmid19127968, year = {2008}, author = {Grandi, G and Morchon, R and Kramer, L and Kartashev, V and Simon, F}, title = {Wolbachia in Dirofilaria repens, an agent causing human subcutaneous dirofilariasis.}, journal = {The Journal of parasitology}, volume = {94}, number = {6}, pages = {1421-1423}, doi = {10.1645/GE-1575.1}, pmid = {19127968}, issn = {0022-3395}, mesh = {Adult ; Animals ; Antibodies, Bacterial/blood ; Bacterial Proteins/immunology ; Dirofilaria/immunology/*microbiology/pathogenicity ; Dirofilariasis/immunology/*parasitology/pathology ; Female ; Granuloma/immunology/*parasitology/pathology ; Humans ; Immunoglobulin G/blood ; Immunohistochemistry ; Male ; Membrane Proteins/immunology ; Middle Aged ; Skin Diseases, Parasitic/immunology/*parasitology/pathology ; Symbiosis ; Wolbachia/immunology/*physiology ; Young Adult ; }, abstract = {Human subcutaneous dirofilariasis is an increasingly reported zoonosis caused by several filarial species, in particular by Dirofilaria (Nochtiella) repens. Like many filarial worms, D. repens harbors the bacterial endosymbiont Wolbachia that has been implicated in the inflammatory features of filarial infection. Immunohistochemical staining against the Wolbachia surface protein (WSP) was carried out on 14 skin nodules and showed numerous bacteria within the intact worms and occasional positive staining within the surrounding inflammatory infiltrate. Serum samples from 11 of these patients resulted positive for total immunoglobulin G titers against WSP as examined in enzyme-linked immunosorbent assay. This is the first description of Wolbachia distribution in D. repens and the first report of specific immune response to Wolbachia in patients with subcutaneous dirofilariasis.}, } @article {pmid19120997, year = {2008}, author = {Gompert, Z and Forister, ML and Fordyce, JA and Nice, CC}, title = {Widespread mito-nuclear discordance with evidence for introgressive hybridization and selective sweeps in Lycaeides.}, journal = {Molecular ecology}, volume = {17}, number = {24}, pages = {5231-5244}, doi = {10.1111/j.1365-294X.2008.03988.x}, pmid = {19120997}, issn = {1365-294X}, mesh = {Amplified Fragment Length Polymorphism Analysis ; Animals ; Butterflies/*genetics/microbiology ; DNA, Mitochondrial/*genetics ; Evolution, Molecular ; Female ; Genes, Mitochondrial ; Genetic Variation ; Genetics, Population ; Haplotypes ; *Hybridization, Genetic ; Inheritance Patterns ; Linkage Disequilibrium ; Male ; Mitochondria/genetics ; North America ; *Selection, Genetic ; Sequence Analysis, DNA ; Wolbachia/physiology ; }, abstract = {We investigated the extent and potential cause(s) of mitochondrial introgression within the polytypic North American Lycaeides species complex (Lepidoptera). By comparing population genetic structure based on mitochondrial DNA (COI and COII) and nuclear DNA (251 polymorphic amplified fragment length polymorphism markers), we detected substantial mito-nuclear discordance, primarily involving a single mitochondrial haplotype (h01), which is likely due to mitochondrial introgression between differentiated Lycaeides populations and/or species. We detected reduced mitochondrial genetic diversity relative to nuclear genetic diversity in populations where mitochondrial haplotype h01 occurs, suggesting that the spread of this haplotype was facilitated by selection. We found no evidence that haplotype h01 is associated with increased fitness (in terms of survival to eclosion, fresh adult weight, and adult longevity) in a polymorphic Lycaeides melissa population. However, we did find a positive association between mitochondrial haplotype h01 and infection by the endoparasitic bacterium Wolbachia in one out of three lineages tested. Linkage disequilibrium between mitochondrial haplotype h01 and Wolbachia infection status may have resulted in indirect selection favouring the spread of haplotype h01 in at least one lineage of North American Lycaeides. These results illustrate the potential for introgressive hybridization to produce substantial mito-nuclear discordance and demonstrate that an individual's mitochondrial and nuclear genome may have strikingly different evolutionary histories resulting from non-neutral processes and intrinsic differences in the inheritance and biology of these genomes.}, } @article {pmid19119237, year = {2009}, author = {McMeniman, CJ and Lane, RV and Cass, BN and Fong, AW and Sidhu, M and Wang, YF and O'Neill, SL}, title = {Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti.}, journal = {Science (New York, N.Y.)}, volume = {323}, number = {5910}, pages = {141-144}, doi = {10.1126/science.1165326}, pmid = {19119237}, issn = {1095-9203}, mesh = {Aedes/genetics/*microbiology/physiology/virology ; Animals ; Blood ; Dengue/transmission ; Dengue Virus/growth & development ; Female ; Humans ; Insect Vectors/genetics/*microbiology/physiology/virology ; Longevity ; Male ; Reproduction ; Symbiosis ; Temperature ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Most pathogens require a relatively long period of development in their mosquito vector before they can be transmitted to a new human host; hence, only older insects are of epidemiological importance. The successful transfer of a life-shortening strain of the inherited bacterial symbiont, Wolbachia, into the major mosquito vector of dengue, Aedes aegypti, halved adult life span under laboratory conditions. The association is stable, and the Wolbachia strain is maternally inherited at high frequency. It is capable of inducing complete cytoplasmic incompatibility, which should facilitate its invasion into natural field populations and its persistence over time. Our data suggest that targeting mosquito age with inherited Wolbachia infections may be a viable strategy to reduce the transmission of pathogens such as dengue viruses.}, } @article {pmid19119208, year = {2009}, author = {Read, AF and Thomas, MB}, title = {Microbiology. Mosquitoes cut short.}, journal = {Science (New York, N.Y.)}, volume = {323}, number = {5910}, pages = {51-52}, doi = {10.1126/science.1168659}, pmid = {19119208}, issn = {1095-9203}, mesh = {Aedes/genetics/*microbiology/physiology/virology ; Animals ; Dengue/prevention & control/transmission ; Dengue Virus/*growth & development ; Female ; Humans ; Insect Vectors/genetics/*microbiology/physiology/virology ; Longevity ; Malaria/prevention & control/transmission ; Male ; Pest Control, Biological ; Selection, Genetic ; Virulence ; Wolbachia/genetics/pathogenicity/*physiology ; }, } @article {pmid20420217, year = {2009}, author = {Machtelinckx, T and Van Leeuwen, T and Vanholme, B and Gehesquière, B and Gheysen, G and De Clercq, P}, title = {Wolbachia affects the reproduction of the mirid predator Macrolophus pygmaeus.}, journal = {Communications in agricultural and applied biological sciences}, volume = {74}, number = {4}, pages = {19-23}, pmid = {20420217}, issn = {1379-1176}, mesh = {Animals ; Female ; Heteroptera/*microbiology/*physiology ; Pest Control, Biological ; Predatory Behavior ; Reproduction ; Wolbachia/*physiology ; }, } @article {pmid19114486, year = {2009}, author = {Salzberg, SL and Puiu, D and Sommer, DD and Nene, V and Lee, NH}, title = {Genome sequence of the Wolbachia endosymbiont of Culex quinquefasciatus JHB.}, journal = {Journal of bacteriology}, volume = {191}, number = {5}, pages = {1725}, pmid = {19114486}, issn = {1098-5530}, support = {R01 GM083873/GM/NIGMS NIH HHS/United States ; R01-LM006845/LM/NLM NIH HHS/United States ; R01 LM006845-09/LM/NLM NIH HHS/United States ; R01 LM006845/LM/NLM NIH HHS/United States ; R01 GM083873-05/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Culex/*microbiology ; *Genome, Bacterial ; Molecular Sequence Data ; *Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Wolbachia species are endosymbionts of a wide range of invertebrates, including mosquitoes, fruit flies, and nematodes. The wPip strains can cause cytoplasmic incompatibility in some strains of the Culex mosquito. Here we describe the genome sequence of a Wolbachia strain that was discovered in the whole-genome sequencing data for the mosquito Culex quinquefasciatus strain JHB.}, } @article {pmid19098217, year = {2009}, author = {Ros, VI and Fleming, VM and Feil, EJ and Breeuwer, JA}, title = {How diverse is the genus Wolbachia? Multiple-gene sequencing reveals a putatively new Wolbachia supergroup recovered from spider mites (Acari: Tetranychidae).}, journal = {Applied and environmental microbiology}, volume = {75}, number = {4}, pages = {1036-1043}, pmid = {19098217}, issn = {1098-5336}, mesh = {Animals ; Bacterial Proteins/genetics ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Evolution, Molecular ; Genes, rRNA ; Molecular Sequence Data ; Phylogeny ; *Polymorphism, Genetic ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Recombination, Genetic ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Tetranychidae/*microbiology ; Wolbachia/*classification/*genetics/isolation & purification ; }, abstract = {At least 20% of all arthropods and some nematode species are infected with intracellular bacteria of the genus Wolbachia. This highly diverse genus has been subdivided into eight "supergroups" (A to H) on the basis of nucleotide sequence data. Here, we report the discovery of a new Wolbachia supergroup recovered from the spider mite species Bryobia species V (Acari: Tetranychidae), based on the sequences of three protein-coding genes (ftsZ, gltA, and groEL) and the 16S rRNA gene. Other tetranychid mites possess supergroup B Wolbachia strains. The discovery of another Wolbachia supergroup expands the known diversity of Wolbachia and emphasizes the high variability of the genus. Our data also clarify the existing supergroup structure and highlight the use of multiple gene sequences for robust phylogenetic analysis. In addition to previous reports of recombination between the arthropod-infecting supergroups A and B, we provide evidence for recombination between the nematode-infecting supergroups C and D. Robust delineation of supergroups is essential for understanding the origin and spread of this common reproductive parasite and for unraveling mechanisms of host adaptation and manipulation across a wide range of hosts.}, } @article {pmid19086265, year = {2008}, author = {Relman, DA}, title = {'Til death do us part': coming to terms with symbiotic relationships. Forward.}, journal = {Nature reviews. Microbiology}, volume = {6}, number = {10}, pages = {721-724}, doi = {10.1038/nrmicro1990}, pmid = {19086265}, issn = {1740-1534}, support = {DP1 OD000964/OD/NIH HHS/United States ; }, mesh = {Animals ; Biodiversity ; Environment ; Humans ; Mycorrhizae/physiology ; RNA, Ribosomal, 16S ; Symbiosis/*physiology ; Vertebrates/microbiology ; Water Microbiology ; Wolbachia/physiology ; }, abstract = {Symbiotic interactions of microorganisms are widespread in nature, and support fundamentally important processes in diverse areas of biology that range from health and disease to ecology and the environment. Here, David Relman discusses the selection of articles in this Focus issue, which reflects the exciting advances in our understanding of intimate partnerships between organisms and their environments.}, } @article {pmid19067840, year = {2008}, author = {Pou-Barreto, C and Quispe-Ricalde, MA and Morchón, R and Vázquez, C and Genchi, M and Postigo, I and Valladares, B and Simón, F}, title = {Galectin and aldolase-like molecules are responsible for the specific IgE response in humans exposed to Dirofilaria immitis.}, journal = {Parasite immunology}, volume = {30}, number = {11-12}, pages = {596-602}, doi = {10.1111/j.1365-3024.2008.01060.x}, pmid = {19067840}, issn = {1365-3024}, mesh = {Aldehyde-Lyases/*immunology ; Animals ; Antibodies, Bacterial/blood ; Antibodies, Helminth/*blood ; Antigens, Helminth/chemistry/*immunology ; Dirofilaria immitis/*immunology/microbiology/physiology ; Dirofilariasis/immunology ; Galectins/*immunology ; Helminth Proteins/chemistry/immunology ; Humans ; Immunoglobulin E/*blood ; Immunoglobulin G/blood ; Lung Diseases/immunology ; Male ; Mass Spectrometry ; Molecular Weight ; Symbiosis/immunology ; Wolbachia/*immunology/physiology ; }, abstract = {Dirofilaria immitis is the agent of the heartworm disease in canids and felids, and of pulmonary dirofilariosis in man. Like other filariae, D. immitis harbours endosymbion Wolbachia bacteriae. In this work we analyse the response of specific IgE antibodies against both D. immitis antigens and the Wolbachia surface protein (WSP) in two groups of persons living in an area of canine endemia, one presenting high levels of total IgE (group 1) and other with normal levels (group 2). Infections with D. immitis were demonstrated by the presence of specific IgG in 228 individuals(48.8%) of the group 1 and only in one of the group 2. Specific IgE antibody response against D. immitis antigens was detected only in individuals of the group 1. IgE response against WSP was not detected in any group. The IgE response was directed mainly against two molecules of 33 and 42 kDa of the antigenic extract of D. immitis. These molecules were identified by mass spectrometry as a galectin and an aldolase, respectively. Their possible role in the survival mechanisms of the parasite and their contribution to development of allergic reactions in individuals resident in areas with heartworm disease are discussed.}, } @article {pmid19063762, year = {2009}, author = {Narita, S and Shimajiri, Y and Nomura, M}, title = {Strong cytoplasmic incompatibility and high vertical transmission rate can explain the high frequencies of Wolbachia infection in Japanese populations of Colias erate poliographus (Lepidoptera: Pieridae).}, journal = {Bulletin of entomological research}, volume = {99}, number = {4}, pages = {385-391}, doi = {10.1017/S0007485308006469}, pmid = {19063762}, issn = {1475-2670}, mesh = {Animals ; Base Sequence ; Butterflies/*microbiology/physiology ; Crosses, Genetic ; *Infectious Disease Transmission, Vertical ; Japan ; Likelihood Functions ; Models, Genetic ; Molecular Sequence Data ; *Phylogeny ; Polymerase Chain Reaction ; Reproduction/physiology ; Sequence Analysis, DNA ; Survival Analysis ; Wolbachia/*genetics ; }, abstract = {Wolbachia, belonging to Alphaproteobacteria, is ubiquitously found in arthropods and filarial nematodes, and is known to manipulate the reproduction of its hosts in various ways, such as feminization, male killing, induction of parthenogenesis or induction of cytoplasmic incompatibility. We found that the Wolbachia infection frequencies of the butterfly Colias erate poliographus were high (85.7-100%) in seven Japanese populations. Crossing experiments and rearing revealed that the Wolbachia strain exhibited strong cytoplasmic incompatibility and perfect vertical transmission in C. erate poliographus. Moreover, a comparison of the survival rates between infected and cured broods suggested that Wolbachia infection had beneficial effects on host fitness. Our findings suggested that the high infection frequencies in Japanese populations have been accomplished by these advantageous traits of the Wolbachia strain. Furthermore, the multilocus sequence typing (MLST) scheme revealed that the Wolbachia in C. erate poliographus is a novel strain (ST141), belonging to supergroup B.}, } @article {pmid19056502, year = {2009}, author = {Linares, MC and Soto-Calderón, ID and Lees, DC and Anthony, NM}, title = {High mitochondrial diversity in geographically widespread butterflies of Madagascar: a test of the DNA barcoding approach.}, journal = {Molecular phylogenetics and evolution}, volume = {50}, number = {3}, pages = {485-495}, doi = {10.1016/j.ympev.2008.11.008}, pmid = {19056502}, issn = {1095-9513}, support = {BBS/B/04358//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Butterflies/classification/*genetics/microbiology ; DNA, Mitochondrial/*genetics ; Electron Transport Complex IV/genetics ; Evolution, Molecular ; Gene Flow ; *Genetic Variation ; Genetics, Population ; Geography ; Mitochondria/genetics ; *Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; Wolbachia/genetics ; }, abstract = {The standardized use of mitochondrial cytochrome c oxidase subunit I (COI) gene sequences as DNA barcodes has been widely promoted as a high-throughput method for species identification and discovery. Species delimitation has been based on the following criteria: (1) monophyletic association and less frequently (2) a minimum 10x greater divergence between than within species. Divergence estimates, however, can be inflated if sister species pairs are not included and the geographic extent of variation within any given taxon is not sampled comprehensively. This paper addresses both potential biases in DNA divergence estimation by sampling range-wide variation in several morphologically distinct, endemic butterfly species in the genus Heteropsis, some of which are sister taxa. We also explored the extent to which mitochondrial DNA from the barcode region can be used to assess the effects of historical rainforest fragmentation by comparing genetic variation across Heteropsis populations with an unrelated forest-associated taxon Saribia tepahi. Unexpectedly, generalized primers led to the inadvertent amplification of the endosymbiont Wolbachia, undermining the use of universal primers and necessitating the design of genus-specific COI primers alongside a Wolbachia-specific PCR assay. Regardless of the high intra-specific genetic variation observed, most species satisfy DNA barcoding criteria and can be differentiated in the nuclear phylogeny. Nevertheless, two morphologically distinguishable candidate species fail to satisfy the barcoding 10x genetic distance criterion, underlining the difficulties of applying a standard distance threshold to species delimitation. Phylogeographic analysis of COI data suggests that forest fragmentation may have played an important role in the recent evolutionary diversification of these butterflies. Further work on other Malagasy taxa using both mitochondrial and nuclear data will provide better insight into the role of historical habitat fragmentation in species diversification and may potentially contribute to the identification of priority areas for conservation.}, } @article {pmid19054050, year = {2009}, author = {Russell, JA and Goldman-Huertas, B and Moreau, CS and Baldo, L and Stahlhut, JK and Werren, JH and Pierce, NE}, title = {Specialization and geographic isolation among Wolbachia symbionts from ants and lycaenid butterflies.}, journal = {Evolution; international journal of organic evolution}, volume = {63}, number = {3}, pages = {624-640}, doi = {10.1111/j.1558-5646.2008.00579.x}, pmid = {19054050}, issn = {1558-5646}, mesh = {Animals ; Ants/*microbiology/physiology ; Bacterial Typing Techniques ; *Biological Evolution ; Butterflies/*microbiology/physiology ; Phylogeny ; Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {Wolbachia are the most prevalent and influential bacteria described among the insects to date. But despite their significance, we lack an understanding of their evolutionary histories. To describe the evolution of symbioses between Wolbachia and their hosts, we surveyed global collections of two diverse families of insects, the ants and lycaenid butterflies. In total, 54 Wolbachia isolates were typed using a Multi Locus Sequence Typing (MLST) approach, in which five unlinked loci were sequenced and analyzed to decipher evolutionary patterns. AMOVA and phylogenetic analyses demonstrated that related Wolbachia commonly infect related hosts, revealing a pattern of host association that was strongest among strains from the ants. A review of the literature indicated that horizontal transfer is most successful when Wolbachia move between related hosts, suggesting that patterns of host association are driven by specialization on a common physiological background. Aside from providing the broadest and strongest evidence to date for Wolbachia specialization, our findings also reveal that strains from New World ants differ markedly from those in ants from other locations. We, therefore, conclude that both geographic and phylogenetic barriers have promoted evolutionary divergence among these influential symbionts.}, } @article {pmid19050762, year = {2008}, author = {Cremer, S and Ugelvig, LV and Drijfhout, FP and Schlick-Steiner, BC and Steiner, FM and Seifert, B and Hughes, DP and Schulz, A and Petersen, KS and Konrad, H and Stauffer, C and Kiran, K and Espadaler, X and d'Ettorre, P and Aktaç, N and Eilenberg, J and Jones, GR and Nash, DR and Pedersen, JS and Boomsma, JJ}, title = {The evolution of invasiveness in garden ants.}, journal = {PloS one}, volume = {3}, number = {12}, pages = {e3838}, pmid = {19050762}, issn = {1932-6203}, mesh = {*Aggression ; Animals ; Ants/chemistry/*genetics/microbiology/parasitology ; Asia ; Beauveria/isolation & purification ; *Behavior, Animal ; *Biological Evolution ; Europe ; *Genetics, Population ; Hydrocarbons/analysis ; Nesting Behavior ; Population Dynamics ; Wolbachia/isolation & purification ; }, abstract = {It is unclear why some species become successful invaders whilst others fail, and whether invasive success depends on pre-adaptations already present in the native range or on characters evolving de-novo after introduction. Ants are among the worst invasive pests, with Lasius neglectus and its rapid spread through Europe and Asia as the most recent example of a pest ant that may become a global problem. Here, we present the first integrated study on behavior, morphology, population genetics, chemical recognition and parasite load of L. neglectus and its non-invasive sister species L. turcicus. We find that L. neglectus expresses the same supercolonial syndrome as other invasive ants, a social system that is characterized by mating without dispersal and large networks of cooperating nests rather than smaller mutually hostile colonies. We conclude that the invasive success of L. neglectus relies on a combination of parasite-release following introduction and pre-adaptations in mating system, body-size, queen number and recognition efficiency that evolved long before introduction. Our results challenge the notion that supercolonial organization is an inevitable consequence of low genetic variation for chemical recognition cues in small invasive founder populations. We infer that low variation and limited volatility in cuticular hydrocarbon profiles already existed in the native range in combination with low dispersal and a highly viscous population structure. Human transport to relatively disturbed urban areas thus became the decisive factor to induce parasite release, a well established general promoter of invasiveness in non-social animals and plants, but understudied in invasive social insects.}, } @article {pmid19043737, year = {2009}, author = {Foster, JM and Raverdy, S and Ganatra, MB and Colussi, PA and Taron, CH and Carlow, CK}, title = {The Wolbachia endosymbiont of Brugia malayi has an active phosphoglycerate mutase: a candidate target for anti-filarial therapies.}, journal = {Parasitology research}, volume = {104}, number = {5}, pages = {1047-1052}, pmid = {19043737}, issn = {1432-1955}, support = {2R44 A1061865-02//PHS HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/*genetics/isolation & purification/*metabolism ; Brugia malayi/*microbiology ; Cloning, Molecular ; Escherichia coli/genetics ; Gene Expression ; Kluyveromyces/genetics ; Molecular Sequence Data ; NAD/metabolism ; Phosphoglycerate Mutase/*genetics/isolation & purification/*metabolism ; Sequence Alignment ; Wolbachia/*enzymology/genetics ; }, abstract = {Phosphoglycerate mutases (PGM) interconvert 2- and 3-phosphoglycerate in the glycolytic and gluconeogenic pathways. A putative cofactor-independent phosphoglycerate mutase gene (iPGM) was identified in the genome sequence of the Wolbachia endosymbiont from the filarial nematode, Brugia malayi (wBm). Since iPGM has no sequence or structural similarity to the cofactor-dependent phosphoglycerate mutase (dPGM) found in mammals, it may represent an attractive Wolbachia drug target. In the present study, wBm-iPGM cloned and expressed in Escherichia coli was mostly insoluble and inactive. However, the protein was successfully produced in the yeast Kluyveromyces lactis and the purified recombinant wBm-iPGM showed typical PGM activity. Our results provide a foundation for further development of wBm-iPGM as a promising new drug target for novel anti-filarial therapies that selectively target the endosymbiont.}, } @article {pmid19036199, year = {2008}, author = {Zhang, X and Pfeiffer, DG}, title = {Evaluation of reproductive compatibility of interstrain matings among plum curculio populations in the eastern United States.}, journal = {Environmental entomology}, volume = {37}, number = {5}, pages = {1208-1213}, doi = {10.1603/0046-225x(2008)37[1208:eorcoi]2.0.co;2}, pmid = {19036199}, issn = {0046-225X}, mesh = {Animals ; *Crosses, Genetic ; Female ; Fertility ; Geography ; Male ; United States ; Weevils/microbiology/*physiology ; Wolbachia/physiology ; }, abstract = {Reproductive compatibility of interstrain matings in plum curculio, Conotrachelus nenuphar (Herbst), was studied during the summers of 2004 and 2006. There was a unidirectional incompatibility in the cross-matings along a rough north to south transect of the range of plum curculio (4 x 4 two-factorial design). There was significantly lower fertility in West Virginia males mated with New York and Virginia females (40 and 29%, respectively), and Florida males had a significantly lower fertility with Virginia and West Virginia females (46 and 37%, respectively), but Florida females were compatible with males of all four populations. Three northern populations were compatible with each other in all cross-mating combinations (New York, Massachusetts, and New Jersey; 3 x 3 two-factorial design). There was a unidirectional reproductive incompatibility in the combination of New Jersey males with Florida females (47%) and a bi-directional incompatibility between Florida and West Virginia reciprocal cross-mating (26 and 21%, respectively). The pattern of the reproductive incompatibility among geographic populations could help us to distinguish plum curculio strains and to clarify their strain distribution range to enrich our knowledge on their population ecology and biosystematics.}, } @article {pmid19028913, year = {2009}, author = {Espino, CI and Gómez, T and González, G and do Santos, MF and Solano, J and Sousa, O and Moreno, N and Windsor, D and Ying, A and Vilchez, S and Osuna, A}, title = {Detection of Wolbachia bacteria in multiple organs and feces of the triatomine insect Rhodnius pallescens (Hemiptera, Reduviidae).}, journal = {Applied and environmental microbiology}, volume = {75}, number = {2}, pages = {547-550}, pmid = {19028913}, issn = {1098-5336}, mesh = {Animal Structures/*microbiology ; Animals ; Feces/*microbiology ; Rhodnius/*microbiology ; Trypanosomatina/isolation & purification ; Wolbachia/*isolation & purification ; }, abstract = {At least two types of Wolbachia bacteria were detected in wild and insectarium-raised Rhodnius pallescens, a natural vector of Trypanosoma cruzi and Trypanosoma rangeli. Wolbachia was detected in all the organs and tissues studied and in the feces, and this provided a methodological advantage for determining the presence of this endosymbiont in this host, obviating the need to kill the specimens. The occurrence of trypanosomatids in wild individuals was also studied.}, } @article {pmid19023073, year = {2008}, author = {Schneider, DS and Chambers, MC}, title = {Microbiology. Rogue insect immunity.}, journal = {Science (New York, N.Y.)}, volume = {322}, number = {5905}, pages = {1199-1200}, doi = {10.1126/science.1167450}, pmid = {19023073}, issn = {1095-9203}, support = {R01 AI069164/AI/NIAID NIH HHS/United States ; T32 AI007328/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Insect Proteins/immunology ; Insecta/*immunology/microbiology ; Peptides/immunology ; Wolbachia/immunology ; }, } @article {pmid19002498, year = {2009}, author = {Singh, M and Srivastava, KK and Bhattacharya, SM}, title = {Molecular cloning and characterization of a novel immunoreactive ATPase/RNA helicase in human filarial parasite Brugia malayi.}, journal = {Parasitology research}, volume = {104}, number = {4}, pages = {753-761}, pmid = {19002498}, issn = {0932-0113}, mesh = {*Adenosine Triphosphatases/chemistry/genetics/metabolism ; Amino Acid Sequence ; Animals ; Antibodies, Helminth/blood ; Brugia malayi/*enzymology/genetics/growth & development/immunology ; *Cloning, Molecular ; Elephantiasis, Filarial/*immunology/parasitology ; Female ; Gene Library ; Humans ; *Immunodominant Epitopes ; Male ; Mice ; Mice, Inbred BALB C ; Microfilariae/*enzymology/genetics/immunology ; Molecular Sequence Data ; *RNA Helicases/chemistry/genetics/metabolism ; Sequence Analysis, DNA ; }, abstract = {DEAD box proteins are putative RNA unwinding proteins found in organisms ranging from mammals to bacteria. We have identified a novel immunodominant cDNA clone, BmL3-helicase, encoding DEAD box RNA helicase by immunoscreening of a larval stage cDNA library of Brugia malayi. The cDNA sequence exhibited strong sequence homology to Caenorhabditis elegans and C. briggsae RNA helicase, a prototypic member of the DEAD (Asp-Glu-Ala-Asp) box protein family. The clone also showed similarity with RNA helicase of Wolbachia, an endosymbiotic bacterium of filarial parasite. It was overexpressed as approximately 50 kDa His-tag fusion protein, and ATP hydrolysis assay of recombinant enzyme showed that either ATP or dATP was required for the unwinding activity, indicating BmL3-helicase as an ATP/dATP-dependent RNA helicase. The recombinant protein also demonstrated cross-seroreactivity with human bancroftian sera. The presence of BmL3-helicase in various life stages of B. malayi was confirmed by immunoblotting of parasite-life-cycle extracts with polyclonal sera against the BmL3-helicase, which showed high levels of expression in microfilaria, L(3,) and adult (both male and female) stages. In the absence of an effective macrofilaricidal agent and validated anti-filarial drug targets, RNA helicases could be utilized as a rational drug target for developing agents against the human filarial parasite.}, } @article {pmid18988686, year = {2009}, author = {Woolfit, M and Iturbe-Ormaetxe, I and McGraw, EA and O'Neill, SL}, title = {An ancient horizontal gene transfer between mosquito and the endosymbiotic bacterium Wolbachia pipientis.}, journal = {Molecular biology and evolution}, volume = {26}, number = {2}, pages = {367-374}, doi = {10.1093/molbev/msn253}, pmid = {18988686}, issn = {1537-1719}, mesh = {Animals ; Culicidae/*genetics/*microbiology/physiology ; Drosophila melanogaster/microbiology ; Gene Transfer, Horizontal ; Phylogeny ; Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {The extent and biological relevance of horizontal gene transfer (HGT) in eukaryotic evolution remain highly controversial. Recent studies have demonstrated frequent and large-scale HGT from endosymbiotic bacteria to their hosts, but the great majority of these transferred genes rapidly become nonfunctional in the recipient genome. Here, we investigate an ancient HGT between a host metazoan and an endosymbiotic bacterium, Wolbachia pipientis. The transferred gene has so far been found only in mosquitoes and Wolbachia. In mosquitoes, it is a member of a gene family encoding candidate receptors required for malaria sporozoite invasion of the mosquito salivary gland. The gene copy in Wolbachia has substantially diverged in sequence from the mosquito homolog, is evolving under purifying selection, and is expressed, suggesting that this gene is also functional in the bacterial genome. Several lines of evidence indicate that the gene may have been transferred from eukaryotic host to bacterial endosymbiont. Regardless of the direction of transfer, however, these results demonstrate that interdomain HGT may give rise to functional, persistent, and possibly evolutionarily significant new genes.}, } @article {pmid18983256, year = {2008}, author = {Moran, NA and McCutcheon, JP and Nakabachi, A}, title = {Genomics and evolution of heritable bacterial symbionts.}, journal = {Annual review of genetics}, volume = {42}, number = {}, pages = {165-190}, doi = {10.1146/annurev.genet.41.110306.130119}, pmid = {18983256}, issn = {0066-4197}, support = {1K 12 GM00708/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteria/*genetics ; Bacterial Physiological Phenomena ; Biological Evolution ; Ecosystem ; Genome, Bacterial ; Genomics ; Insecta/*microbiology ; Symbiosis/*genetics ; }, abstract = {Insect heritable symbionts have proven to be ubiquitous, based on molecular screening of various insect lineages. Recently, molecular and experimental approaches have yielded an immensely richer understanding of their diverse biological roles, resulting in a burgeoning research literature. Increasingly, commonalities and intermediates are being discovered between categories of symbionts once considered distinct: obligate mutualists that provision nutrients, facultative mutualists that provide protection against enemies or stress, and symbionts such as Wolbachia that manipulate reproductive systems. Among the most far-reaching impacts of widespread heritable symbiosis is that it may promote speciation by increasing reproductive and ecological isolation of host populations, and it effectively provides a means for transfer of genetic information among host lineages. In addition, insect symbionts provide some of the extremes of cellular genomes, including the smallest and the fastest evolving, raising new questions about the limits of evolution of life.}, } @article {pmid18979132, year = {2009}, author = {Fernando de Souza, R and Daivison Silva Ramalho, J and Santina de Castro Morini, M and Wolff, JL and Araújo, RC and Mascara, D}, title = {Identification and characterization of Wolbachia in Solenopsis saevissima fire ants (Hymenoptera: Formicidae) in southeastern Brazil.}, journal = {Current microbiology}, volume = {58}, number = {3}, pages = {189-194}, pmid = {18979132}, issn = {1432-0991}, mesh = {Animals ; Ants/*microbiology ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; Brazil ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {The genus Solenopsis appears to have evolved and radiated very rapidly in South America and then spread throughout the rest of the continent. As part of the expansion process, distribution patterns and different degrees of geographic isolation among populations of S. saevissima can be observed. We have investigated the presence of Wolbachia in 52 colonies and 1623 individuals in southeastern Brazil. Detection of Wolbachia infection was based on amplification of the 16S rRNA and wsp genes by polymerase chain reactions. Wolbachia was found in only one of the four locations investigated and it was observed that the populations were polymorphic for infection. The infection level observed increased during the period of screening. In particular, double infection (16SWA and B) increased from 44% in 2005 to 90% in 2006. The A-group of Wolbachia from the wsp sequences was determined by sequencing. However, two variant wsp sequences were detected in Wolbachia present in these populations. The alignment of our sequences with those deposited in GenBank indicated significant differences in relation to homologous sequences. Phylogenetic relationships were inferred using parsimony, and confidence intervals were estimated by bootstrapping. Then the divergence of the Wolbachia of S. saevissima in the populations studied with other variants allowed us to verify that wSS1 and dwSS2 formed a distinct clade within the A-group (>75%). These results can be useful in studies on the dynamics of ant populations.}, } @article {pmid18979023, year = {2007}, author = {Rasgon, J}, title = {Population replacement strategies for controlling vector populations and the use of Wolbachia pipientis for genetic drive.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {5}, pages = {225}, doi = {10.3791/225}, pmid = {18979023}, issn = {1940-087X}, mesh = {Animals ; Dengue/prevention & control ; Disease Transmission, Infectious/*prevention & control ; Humans ; Immunity, Innate/*genetics ; Insect Vectors/*microbiology ; Malaria/prevention & control ; Population Control/*methods ; *Population Dynamics ; Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {In this video, Jason Rasgon discusses population replacement strategies to control vector-borne diseases such as malaria and dengue. "Population replacement" is the replacement of wild vector populations (that are competent to transmit pathogens) with those that are not competent to transmit pathogens. There are several theoretical strategies to accomplish this. One is to exploit the maternally-inherited symbiotic bacteria Wolbachia pipientis. Wolbachia is a widespread reproductive parasite that spreads in a selfish manner at the extent of its host's fitness. Jason Rasgon discusses, in detail, the basic biology of this bacterial symbiont and various ways to use it for control of vector-borne diseases.}, } @article {pmid18979021, year = {2007}, author = {Gamston, C and Rasgon, J}, title = {Maintaining Wolbachia in cell-free medium.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {5}, pages = {223}, doi = {10.3791/223}, pmid = {18979021}, issn = {1940-087X}, mesh = {Animals ; Bacteriological Techniques ; Cell Division ; *Cell-Free System ; Cells, Cultured ; Culicidae/cytology/microbiology ; *Culture Media ; Microbial Viability ; *Symbiosis ; Wolbachia/cytology/*isolation & purification/*physiology ; }, abstract = {In this video protocol, procedures are demonstrated to (1) purify Wolbachia symbionts out of cultured mosquito cells, (2) use a fluorescent assay to ascertain the viability of the purified Wolbachia and (3) maintain the now extracellular Wolbachia in cell-free medium. Purified Wolbachia remain alive in the extracellular phase but do not replicate until re-inoculated into eukaryotic cells. Extracellular Wolbachia purified in this manner will remain viable for at least a week at room temperature, and possibly longer. Purified Wolbachia are suitable for micro-injection, DNA extraction and other applications.}, } @article {pmid18978537, year = {2008}, author = {Hoerauf, A}, title = {Filariasis: new drugs and new opportunities for lymphatic filariasis and onchocerciasis.}, journal = {Current opinion in infectious diseases}, volume = {21}, number = {6}, pages = {673-681}, doi = {10.1097/QCO.0b013e328315cde7}, pmid = {18978537}, issn = {1473-6527}, mesh = {Animals ; Doxycycline/administration & dosage/*therapeutic use ; Elephantiasis, Filarial/*drug therapy ; Filaricides/administration & dosage/*therapeutic use ; Humans ; Onchocerca volvulus/drug effects ; Onchocerciasis/*drug therapy ; Wuchereria bancrofti/drug effects ; }, abstract = {PURPOSE OF REVIEW: Chemotherapy against onchocerciasis and lymphatic filariasis has been discussed mainly within the framework of mass drug administration with diethylcarbamazine, ivermectin and albendazole. Although strong reductions in infection prevalence were achieved, the regimes for these drugs do not fully cover needs of individual patients who seek treatment because of symptoms. Chemotherapy against filarial Wolbachia endosymbionts with doxycycline showed higher antiparasitic efficacy in onchocerciasis and lymphatic filariasis and also improved disease. This review details the recent indications for this new treatment, focussing on regimes for individual drug administration.

RECENT FINDINGS: A regimen of 6-week/100 mg doxycycline per day sterilized adult female Onchocerca volvulus. Two hundred milligrams doxycycline per day for 4 or 6 weeks revealed 50 and 60% macrofilaricidal effects, respectively. Reduction of 80-90% of adult worms was observed in bancroftian filariasis with 200 mg/day doxycycline for 4 or 6 weeks. The latter regimen showed reduction of lymph vessel dilation and of hydrocele. Lymphoedema progression was halted and reversed in early stages.

SUMMARY: Different options for antiwolbachial individual drug administration are summarized here. With improving health systems in endemic countries, individuals will demand best-possible treatment and accelerate a shift from mass drug administration-only approaches to integrated approaches combining both mass drug administration and individual drug administration. Treatment may be further improved by new antibiotics detected in high-throughput settings.}, } @article {pmid18977224, year = {2009}, author = {Rao, RU and Huang, Y and Fischer, K and Fischer, PU and Weil, GJ}, title = {Brugia malayi: Effects of nitazoxanide and tizoxanide on adult worms and microfilariae of filarial nematodes.}, journal = {Experimental parasitology}, volume = {121}, number = {1}, pages = {38-45}, doi = {10.1016/j.exppara.2008.09.020}, pmid = {18977224}, issn = {1090-2449}, mesh = {Animals ; Antiparasitic Agents/*pharmacology/therapeutic use ; Brugia malayi/*drug effects/growth & development/ultrastructure ; Dose-Response Relationship, Drug ; Female ; Filariasis/*drug therapy/parasitology ; Gerbillinae ; Male ; Microfilariae/drug effects/growth & development/ultrastructure ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Nitro Compounds ; Thiazoles/*pharmacology/therapeutic use ; }, abstract = {There is an urgent need for safe and effective antifilarials. Prior studies have shown that the nitazoxanide (NTZ) exhibits broad activity against anaerobic bacteria, protozoa, and certain intestinal helminths. We examined the effects of NTZ and tizoxanide (TZ) on Brugia malayi nematodes in vitro and in vivo. In vitro, NTZ and TZ reduced worm motility and viability in a dose-dependent manner. Worm viability was reduced by 50% with both compounds at 2.5 and 20 microg/ml killed adult worms. NTZ or TZ (5 microg/ml) significantly reduced microfilaria release. These compounds blocked worm's embryogenesis, and decreased microfilarial motility and viability. Treated worms had damaged cuticles and abnormal mitochondria. Wolbachia were not cleared by NTZ or TZ treatment. Neither NTZ nor TZ cleared adult worms or microfilariae in infected gerbils. These results show that NTZ and TZ have potent effects on B. malayi nematodes in vitro. However, they were not effective in vivo.}, } @article {pmid18974344, year = {2008}, author = {Hedges, LM and Brownlie, JC and O'Neill, SL and Johnson, KN}, title = {Wolbachia and virus protection in insects.}, journal = {Science (New York, N.Y.)}, volume = {322}, number = {5902}, pages = {702}, doi = {10.1126/science.1162418}, pmid = {18974344}, issn = {1095-9203}, mesh = {Animals ; Drosophila melanogaster/*microbiology/physiology/*virology ; Host-Pathogen Interactions ; Insect Viruses/*pathogenicity ; Nodaviridae/pathogenicity ; RNA Viruses/*pathogenicity ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis bacteria are common endosymbionts of insects that are best known for their ability to increase their prevalence in populations by manipulating host reproductive systems. However, there are examples of Wolbachia that exist in nature that seem to induce no reproductive parasitism trait and yet are able to invade populations. We demonstrate a fitness benefit for Wolbachia-infected insects that may explain this paradox. Drosophila melanogaster flies infected with Wolbachia are less susceptible to mortality induced by a range of RNA viruses. The antiviral protection associated with Wolbachia infection might be exploited in future strategies to reduce transmission of pathogens by insects.}, } @article {pmid18974066, year = {2009}, author = {Bordenstein, SR and Paraskevopoulos, C and Dunning Hotopp, JC and Sapountzis, P and Lo, N and Bandi, C and Tettelin, H and Werren, JH and Bourtzis, K}, title = {Parasitism and mutualism in Wolbachia: what the phylogenomic trees can and cannot say.}, journal = {Molecular biology and evolution}, volume = {26}, number = {1}, pages = {231-241}, pmid = {18974066}, issn = {1537-1719}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R01 GM085163-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/microbiology ; Bayes Theorem ; Filarioidea/microbiology ; Phylogeny ; Symbiosis ; Wolbachia/*classification/genetics/*physiology ; }, abstract = {Ecological and evolutionary theories predict that parasitism and mutualism are not fixed endpoints of the symbiotic spectrum. Rather, parasitism and mutualism may be host or environment dependent, induced by the same genetic machinery, and shifted due to selection. These models presume the existence of genetic or environmental variation that can spur incipient changes in symbiotic lifestyle. However, for obligate intracellular bacteria whose genomes are highly reduced, studies specify that discrete symbiotic associations can be evolutionarily stable for hundreds of millions of years. Wolbachia is an inherited obligate, intracellular infection of invertebrates containing taxa that act broadly as both parasites in arthropods and mutualists in certain roundworms. Here, we analyze the ancestry of mutualism and parasitism in Wolbachia and the evolutionary trajectory of this variation in symbiotic lifestyle with a comprehensive, phylogenomic analysis. Contrary to previous claims, we show unequivocally that the transition in lifestyle cannot be reconstructed with current methods due to long-branch attraction (LBA) artifacts of the distant Anaplasma and Ehrlichia outgroups. Despite the use of 1) site-heterogenous phylogenomic methods that can overcome systematic error, 2) a taxonomically rich set of taxa, and 3) statistical assessments of the genes, tree topologies, and models of evolution, we conclude that the LBA artifact is serious enough to afflict past and recent claims including the root lies in the middle of the Wolbachia mutualists and parasites. We show that different inference methods yield different results and high bootstrap support did not equal phylogenetic accuracy. Recombination was rare among this taxonomically diverse data set, indicating that elevated levels of recombination in Wolbachia are restricted to specific coinfecting groups. In conclusion, we attribute the inability to root the tree to rate heterogeneity between the ingroup and outgroup. Site-heterogenous models of evolution did improve the placement of aberrant taxa in the ingroup phylogeny. Finally, in the unrooted topology, the distribution of parasitism and mutualism across the tree suggests that at least two interphylum transfers shaped the origins of nematode mutualism and arthropod parasitism. We suggest that the ancestry of mutualism and parasitism is not resolvable without more suitable outgroups or complete genome sequences from all Wolbachia supergroups.}, } @article {pmid18973689, year = {2008}, author = {Lohman, DJ and Peggie, D and Pierce, NE and Meier, R}, title = {Phylogeography and genetic diversity of a widespread Old World butterfly, Lampides boeticus (Lepidoptera: Lycaenidae).}, journal = {BMC evolutionary biology}, volume = {8}, number = {}, pages = {301}, pmid = {18973689}, issn = {1471-2148}, mesh = {Animals ; Cytochromes b/genetics ; Electron Transport Complex IV/genetics ; *Genetic Variation ; Genetics, Population ; Haplotypes/genetics ; Lepidoptera/*classification/*genetics ; *Phylogeny ; }, abstract = {BACKGROUND: Evolutionary genetics provides a rich theoretical framework for empirical studies of phylogeography. Investigations of intraspecific genetic variation can uncover new putative species while allowing inference into the evolutionary origin and history of extant populations. With a distribution on four continents ranging throughout most of the Old World, Lampides boeticus (Lepidoptera: Lycaenidae) is one of the most widely distributed species of butterfly. It is placed in a monotypic genus with no commonly accepted subspecies. Here, we investigate the demographic history and taxonomic status of this widespread species, and screen for the presence or absence of the bacterial endosymbiont Wolbachia.

RESULTS: We performed phylogenetic, population genetic, and phylogeographic analyses using 1799 bp of mitochondrial sequence data from 57 specimens collected throughout the species' range. Most of the samples (>90%) were nearly genetically identical, with uncorrected pairwise sequence differences of 0-0.5% across geographic distances >9,000 km. However, five samples from central Thailand, Madagascar, northern Australia and the Moluccas formed two divergent clades differing from the majority of samples by uncorrected pairwise distances ranging from 1.79-2.21%. Phylogenetic analyses suggest that L. boeticus is almost certainly monophyletic, with all sampled genes coalescing well after the divergence from three closely related taxa included for outgroup comparisons. Analyses of molecular diversity indicate that most L. boeticus individuals in extant populations are descended from one or two relatively recent population bottlenecks.

CONCLUSION: The combined analyses suggest a scenario in which the most recent common ancestor of L. boeticus and its sister taxon lived in the African region approximately 7 Mya; extant lineages of L. boeticus began spreading throughout the Old World at least 1.5 Mya. More recently, expansion after population bottlenecks approximately 1.4 Mya seem to have displaced most of the ancestral polymorphism throughout its range, though at least two early-branching lineages still persist. One of these lineages, in northern Australia and the Moluccas, may have experienced accelerated differentiation due to infection with the bacterial endosymbiont Wolbachia, which affects reproduction. Examination of a haplotype network suggests that Australia has been colonized by the species several times. While there is little evidence for the existence of morphologically cryptic species, these results suggest a complex history affected by repeated dispersal events.}, } @article {pmid18952001, year = {2009}, author = {Scott, AL and Ghedin, E}, title = {The genome of Brugia malayi - all worms are not created equal.}, journal = {Parasitology international}, volume = {58}, number = {1}, pages = {6-11}, pmid = {18952001}, issn = {1383-5769}, support = {KL2 RR025006/RR/NCRR NIH HHS/United States ; KL2 RR025006-02/RR/NCRR NIH HHS/United States ; U01 HL066623/HL/NHLBI NIH HHS/United States ; U01 HL066623-04/HL/NHLBI NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*genetics/growth & development/microbiology ; Caenorhabditis/genetics/growth & development ; Filariasis/parasitology ; *Genome ; Genomics ; Host-Pathogen Interactions ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA ; Wolbachia/growth & development ; }, abstract = {Filarial nematode parasites, the causative agents of elephantiasis and river blindness, undermine the livelihoods of over one hundred million people in the developing world. Recently, the Filarial Genome Project reported the draft sequence of the ~95 Mb genome of the human filarial parasite Brugia malayi - the first parasitic nematode genome to be sequenced. Comparative genome analysis with the prevailing model nematode Caenorhabditis elegans revealed similarities and differences in genome structure and organization that will prove useful as additional nematode genomes are completed. The Brugia genome provides the first opportunity to comprehensively compare the full gene repertoire of a free-living nematode species and one that has evolved as a human pathogen. The Brugia genome also provides an opportunity to gain insight into genetic basis for mutualism, as Brugia, like a majority of filarial species, harbors an endosybiotic bacterium (Wolbachia). The goal of this review is to provide an overview of the results of genomic analysis and how these observations provide new insights into the biology of filarial species.}, } @article {pmid18947926, year = {2008}, author = {Kramer, L and Grandi, G and Leoni, M and Passeri, B and McCall, J and Genchi, C and Mortarino, M and Bazzocchi, C}, title = {Wolbachia and its influence on the pathology and immunology of Dirofilaria immitis infection.}, journal = {Veterinary parasitology}, volume = {158}, number = {3}, pages = {191-195}, doi = {10.1016/j.vetpar.2008.09.014}, pmid = {18947926}, issn = {0304-4017}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Arsenicals/therapeutic use ; Bacterial Outer Membrane Proteins/immunology ; Dirofilaria immitis/*microbiology/pathogenicity ; Dirofilariasis/immunology/microbiology/parasitology/*pathology ; Dog Diseases/immunology/microbiology/parasitology/*pathology ; Dogs ; Doxycycline/therapeutic use ; Female ; Filaricides/*therapeutic use ; Immunohistochemistry/veterinary ; Ivermectin/therapeutic use ; Lung/parasitology/*pathology ; Male ; Severity of Illness Index ; Triazines/therapeutic use ; Wolbachia/drug effects/*immunology ; }, abstract = {Since the definitive identification in 1995 of the bacterial endosymbiont Wolbachia that resides in different tissues of the filarial worm Dirofilaria immitis, there has been increasing interest to understand whether and what role it plays in the pathogenesis of and immune response to heartworm infection. The present study evaluated the effects of treatments on lung pathology in 20 beagle dogs experimentally infected with D. immitis. Dogs in Group 1 were treated with doxycycline (10 mg/kg/day) orally from weeks 0-6, 10-12, 16-18, 22-26, and 28-34. Dogs in Group 2 served as infected, non-treated controls. Dogs in Group 3 were given doxycycline as described for Group 1 combined with weekly oral doses of ivermectin (6 mcg/kg) for 34 weeks and intramuscular (IM) melarsomine (2.5 mg/kg) at week 24, followed by two additional melarsomine injections 24h apart 1 month later. Group 4 received only melarsomine as described for Group 3. Lung lesion criteria, scored by two independent blinded pathologists, included perivascular inflammation and endothelial proliferation. Doxycycline treatment alone had no effect on lesion scores, whereas the combination of doxycycline and ivermectin resulted in less severe perivascular inflammation. All lungs were evaluated for positive immunostaining for the Wolbachia surface protein (WSP). Control dogs showed numerous thrombi, intense perivascular and interstitial inflammation and, occasionally, positive staining for WSP. Interestingly, dogs receiving doxycycline/ivermectin/melarsomine showed significantly less severe arterial lesions and the virtual absence of thrombi.}, } @article {pmid18941470, year = {2008}, author = {Duron, O}, title = {Insights beyond Wolbachia-Drosophila interactions: never completely trust a model: insights from cytoplasmic incompatibility beyond Wolbachia-Drosophila interactions.}, journal = {Heredity}, volume = {101}, number = {6}, pages = {473-474}, doi = {10.1038/hdy.2008.113}, pmid = {18941470}, issn = {1365-2540}, mesh = {Animals ; Drosophila/cytology/*microbiology ; Female ; *Host-Pathogen Interactions ; Male ; Wolbachia/*physiology ; }, } @article {pmid18931357, year = {2008}, author = {Lee, SA and Lee, SG and Choi, EJ and Hyun, C}, title = {Prevalence of the endosymbiont Wolbachia in heartworms (Dirofilaria immitis).}, journal = {The Veterinary record}, volume = {163}, number = {16}, pages = {484-486}, doi = {10.1136/vr.163.16.484}, pmid = {18931357}, issn = {0042-4900}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics/*isolation & purification ; DNA, Bacterial/analysis ; DNA, Helminth/analysis ; Dirofilaria immitis/genetics/isolation & purification/*microbiology ; Dirofilariasis/*parasitology ; Dog Diseases/*parasitology ; Dogs ; Female ; Korea ; Male ; Polymerase Chain Reaction ; Prevalence ; *Symbiosis ; Wolbachia/genetics/*isolation & purification/physiology ; }, } @article {pmid18930598, year = {2008}, author = {McCall, JW and Genchi, C and Kramer, L and Guerrero, J and Dzimianski, MT and Supakorndej, P and Mansour, AM and McCall, SD and Supakorndej, N and Grandi, G and Carson, B}, title = {Heartworm and Wolbachia: therapeutic implications.}, journal = {Veterinary parasitology}, volume = {158}, number = {3}, pages = {204-214}, doi = {10.1016/j.vetpar.2008.09.008}, pmid = {18930598}, issn = {0304-4017}, mesh = {Aedes/microbiology ; Animals ; Anti-Bacterial Agents/adverse effects/therapeutic use ; Antiparasitic Agents/adverse effects/therapeutic use ; Arsenicals/adverse effects/*therapeutic use ; Dirofilaria immitis/drug effects/*microbiology ; Dirofilariasis/*drug therapy ; Dog Diseases/drug therapy ; Dogs ; Dose-Response Relationship, Drug ; Doxycycline/adverse effects/*therapeutic use ; Female ; Filaricides/adverse effects/*therapeutic use ; Ivermectin/adverse effects/*therapeutic use ; Male ; Microfilariae ; Random Allocation ; Thromboembolism/chemically induced/prevention & control/veterinary ; Time Factors ; Treatment Outcome ; Triazines/adverse effects/*therapeutic use ; Wolbachia/drug effects ; }, abstract = {A safer, more effective adulticidal treatment and a safe method for reducing microfilaremia and breaking transmission of heartworm disease early in the treatment are needed. The present study evaluated efficacy of ivermectin (IVM) and doxycycline (DOXY) alone or together (with or without melarsomine [MEL]) in dogs with induced adult heartworm infection and assessed the ability of microfilariae from DOXY-treated dogs to develop to L3 in Aedes aegypti mosquitoes and subsequently to become reproductive adults in dogs. Thirty beagles were each infected with 16 adult heartworms by intravenous transplantation. Six weeks later, dogs were ranked by microfilarial count and randomly allocated to 6 groups of 5 dogs each. Beginning on Day 0, Group 1 received IVM (6 mcg/kg) weekly for 36 weeks. Group 2 received DOXY (10 mcg/(kgday)) orally Weeks 1-6, 10-11, 16-17, 22-25, and 28-33. Groups 3 and 5 received IVM and DOXY according to doses and schedules used for Groups 1 and 2. At Week 24, Groups 3 and 4 received an intramuscular injection of MEL (2.5 mg/kg), followed 1 month later by two injections 24h apart. Group 6 was not treated. Blood samples were collected for periodic microfilaria counts and antigen (Ag) testing (and later immunologic evaluation and molecular biology procedures). Radiographic and physical examinations, hematology/clinical chemistry testing, and urinalysis were done before infection, before Day 0, and periodically during the treatment period. At 36 weeks, the dogs were euthanized and necropsied for worm recovery, collection of lung, liver, kidney, and spleen samples for examination by immunohistochemistry and conventional histological methods. All dogs treated with IVM + DOXY (with or without MEL) were amicrofilaremic after Week 9. Microfilarial counts gradually decreased in dogs treated with IVM or DOXY, but most had a few microfilariae at necropsy. Microfilarial counts for dogs treated only with MEL were similar to those for controls. Antigen test scores gradually decreased with IVM + DOXY (with or without MEL) and after MEL. Antigen scores for IVM or DOXY alone were similar to controls throughout the study. Reduction of adult worms was 20.3% for IVM, 8.7% for DOXY, 92.8% for IVM + DOXY + MEL, 100% for MEL, and 78.3% for IVM + DOXY. Mosquitoes that fed on blood from DOXY-treated dogs had L3 normal in appearance but were not infective for dogs. Preliminary observations suggest that administration of DOXY+IVM for several months prior to (or without) MEL will eliminate adult HW with less potential for severe thromboembolism than MEL alone.}, } @article {pmid18922634, year = {2008}, author = {Simón, F and Morchón, R and Rodríguez-Barbero, A and López-Belmonte, J and Grandi, G and Genchi, C}, title = {Dirofilaria immitis and Wolbachia-derived antigens: its effect on endothelial mammal cells.}, journal = {Veterinary parasitology}, volume = {158}, number = {3}, pages = {223-231}, doi = {10.1016/j.vetpar.2008.09.010}, pmid = {18922634}, issn = {0304-4017}, mesh = {Animals ; Antigens, Helminth/*immunology ; Cells, Cultured ; Dirofilaria immitis/*immunology ; Endothelial Cells/immunology/metabolism ; Endothelium, Vascular/*cytology/immunology/metabolism ; Humans ; Inflammation/immunology ; Intercellular Adhesion Molecule-1/genetics/immunology/metabolism ; Membrane Proteins/*immunology ; Platelet Endothelial Cell Adhesion Molecule-1/genetics/immunology/metabolism ; Vascular Cell Adhesion Molecule-1/genetics/immunology/metabolism ; Vascular Endothelial Growth Factor A/genetics/immunology/metabolism ; Wolbachia/*immunology ; }, abstract = {Antigens of both Dirofilaria immitis and Wolbachia symbiont bacteria are implicated in the inflammatory pathology of heartworm infection. The aim of the present study was to compare the stimulatory capacity of in vitro cultures of vascular endothelial cells by the adult somatic antigens of D. immitis (DiSA) and the recombinant form of the Wolbachia surface protein (rWSP), during the first 24h of stimulation. Our results indicate a different stimulatory activity of the two antigens. Both the DiSA and rWSP stimulate the production of the enzymes responsible of the arachidonic acid metabolism, cyclooxygenase-2, 5-lipoxygenase (5-LO), and leukotriene B4. Only DiSA stimulates the production of prostaglandin E2. Related to the adhesion molecules, the DiSA stimulates the expression of intercellular adhesion molecule-1 (ICAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1), whereas rWSP stimulates ICAM-1, PECAM-1, and vascular cell adhesion molecule-1 (VCAM-1). Expression of E-cadherin and vascular endothelial growth factor also were stimulated by rWSP. Neither of the two antigens altered the basic physiological mechanisms of endothelial cells, such as cell proliferation, cell cycle, or apoptosis. The biological and pathological significance of these finding are discussed.}, } @article {pmid18850111, year = {2009}, author = {Hoerauf, A and Specht, S and Marfo-Debrekyei, Y and Büttner, M and Debrah, AY and Mand, S and Batsa, L and Brattig, N and Konadu, P and Bandi, C and Fimmers, R and Adjei, O and Büttner, DW}, title = {Efficacy of 5-week doxycycline treatment on adult Onchocerca volvulus.}, journal = {Parasitology research}, volume = {104}, number = {2}, pages = {437-447}, pmid = {18850111}, issn = {0932-0113}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/administration & dosage/*therapeutic use ; Doxycycline/administration & dosage/*therapeutic use ; Female ; Filaricides/administration & dosage/*therapeutic use ; Ghana ; Humans ; Male ; Middle Aged ; Onchocerca volvulus/*microbiology/*physiology ; Onchocerciasis/*drug therapy ; Skin/parasitology/pathology ; Survival Analysis ; Wolbachia/*drug effects ; Young Adult ; }, abstract = {The effects of 5-week doxycycline treatment on the depletion of Wolbachia endobacteria from Onchocerca volvulus, on the interruption of embryogenesis and on microfilariae production, and with regard to macrofilaricidal activity were studied. In 2003, in an endemic area in Ghana, 22 onchocerciasis patients received 100 mg/day doxycycline for 5 weeks. Two years after the start of the study, 20 treated and ten untreated patients were nodulectomized and skin microfilariae were counted. The onchocercomas were examined by immunohistology for the presence of Wolbachia, embryogenesis, and vitality of adult filariae. The latter two parameters were further assessed by alternating logistic regression analysis, taking into account the dependency of worms and nodules in patients. Doxycycline resulted in depletion of Wolbachia and in complete interruption of embryogenesis in all worms that were assumed to have been present during treatment. In the treated patients, only 51% of the female worms were alive, compared to 84% in the untreated patients, indicating a moderate but distinct macrofilaricidal activity of doxycycline at this dose. It is concluded that, in areas with ongoing transmission, doxycycline cannot replace regular ivermectin mass treatment because new infections would require repeated rounds of doxycycline. However, doxycycline can be used for the treatment of individuals outside transmission areas, in foci where ivermectin resistance may occur, and in countries where onchocerciasis and loiasis are co-endemic.}, } @article {pmid18836146, year = {2009}, author = {Walker, T and Song, S and Sinkins, SP}, title = {Wolbachia in the Culex pipiens group mosquitoes: introgression and superinfection.}, journal = {The Journal of heredity}, volume = {100}, number = {2}, pages = {192-196}, pmid = {18836146}, issn = {1465-7333}, support = {/WT_/Wellcome Trust/United Kingdom ; 079059/WT_/Wellcome Trust/United Kingdom ; GR079059MA/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Crosses, Genetic ; Culex/classification/*microbiology/physiology ; Embryo, Nonmammalian ; Female ; Genetic Variation/physiology ; Host-Pathogen Interactions/genetics ; Hybridization, Genetic/*physiology ; Male ; Population Dynamics ; Rickettsiaceae Infections/*genetics/microbiology ; Superinfection/*genetics/microbiology ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia bacteria in mosquitoes induce cytoplasmic incompatibility (CI), where sperm from Wolbachia-infected males can produce inviable progeny. The wPip strain in the Culex pipiens group of mosquitoes produces a complexity of CI crossing types. Several factors are thought to be capable of influencing the expression of CI including Wolbachia strain type and host genotype. In this study, the unidirectional CI that occurs between 2 C. pipiens complex laboratory strains, Col and Mol, was further investigated by nuclear genotype introgression. The unidirectional CI between Col and Mol was not found to be influenced by host genetic background, in contrast to a previous introgression study carried out using bidirectionally incompatible C. pipiens group strains. A line containing both wPip strain variants superinfection was also generated by embryonic cytoplasmic transfer. The same crossing type as the parental Col strain was observed in the superinfected line. Quantitative polymerase chain reaction demonstrated a low density of the injected wPipMol variant in the superinfected line after 18 generations, which was considered likely to be responsible for the crossing patterns observed. The Wolbachia density was also shown to be lower in the parental Mol strain males compared with Col strain males, and no inverse relationship between WO phage and Wolbachia density could be detected.}, } @article {pmid18836024, year = {2008}, author = {McMeniman, CJ and Lane, AM and Fong, AW and Voronin, DA and Iturbe-Ormaetxe, I and Yamada, R and McGraw, EA and O'Neill, SL}, title = {Host adaptation of a Wolbachia strain after long-term serial passage in mosquito cell lines.}, journal = {Applied and environmental microbiology}, volume = {74}, number = {22}, pages = {6963-6969}, pmid = {18836024}, issn = {1098-5336}, mesh = {*Adaptation, Biological ; Aedes ; Animals ; Anopheles ; Cell Line ; Drosophila melanogaster/microbiology ; Female ; Gram-Negative Bacterial Infections/microbiology ; Male ; *Serial Passage ; Virulence ; Wolbachia/pathogenicity/*physiology ; }, abstract = {The horizontal transfer of the bacterium Wolbachia pipientis between invertebrate hosts hinges on the ability of Wolbachia to adapt to new intracellular environments. The experimental transfer of Wolbachia between distantly related host species often results in the loss of infection, presumably due to an inability of Wolbachia to adapt quickly to the new host. To examine the process of adaptation to a novel host, we transferred a life-shortening Wolbachia strain, wMelPop, from the fruit fly Drosophila melanogaster into a cell line derived from the mosquito Aedes albopictus. After long-term serial passage in this cell line, we transferred the mosquito-adapted wMelPop into cell lines derived from two other mosquito species, Aedes aegypti and Anopheles gambiae. After a prolonged period of serial passage in mosquito cell lines, wMelPop was reintroduced into its native host, D. melanogaster, by embryonic microinjection. The cell line-adapted wMelPop strains were characterized by a loss of infectivity when reintroduced into the original host, grew to decreased densities, and had reduced abilities to cause life-shortening infection and cytoplasmic incompatibility compared to the original strain. We interpret these shifts in phenotype as evidence for genetic adaptation to the mosquito intracellular environment. The use of cell lines to preadapt Wolbachia to novel hosts is suggested as a possible strategy to improve the success of transinfection in novel target insect species.}, } @article {pmid18830425, year = {2007}, author = {Frydman, H}, title = {Wolbachia bacterial infection in Drosophila.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {2}, pages = {158}, doi = {10.3791/158}, pmid = {18830425}, issn = {1940-087X}, mesh = {Animals ; Drosophila/*microbiology ; *Rickettsiaceae Infections ; *Wolbachia ; }, } @article {pmid18826448, year = {2009}, author = {Raychoudhury, R and Baldo, L and Oliveira, DC and Werren, JH}, title = {Modes of acquisition of Wolbachia: horizontal transfer, hybrid introgression, and codivergence in the Nasonia species complex.}, journal = {Evolution; international journal of organic evolution}, volume = {63}, number = {1}, pages = {165-183}, doi = {10.1111/j.1558-5646.2008.00533.x}, pmid = {18826448}, issn = {1558-5646}, mesh = {Animals ; *Biological Evolution ; DNA, Mitochondrial/genetics ; Phylogeny ; Wasps/classification/genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia are maternally inherited bacteria that infect a large number of insects and are responsible for different reproductive alterations of their hosts. One of the key features of Wolbachia biology is its ability to move within and between host species, which contributes to the impressive diversity and range of infected hosts. Using multiple Wolbachia genes, including five developed for Multi-Locus Sequence Typing (MLST), the diversity and modes of movement of Wolbachia within the wasp genus Nasonia were investigated. Eleven different Wolbachia were found in the four species of Nasonia, including five newly identified infections. Five infections were acquired by horizontal transmission from other insect taxa, three have been acquired by hybridization between two Nasonia species, which resulted in a mitochondrial-Wolbachia sweep from one species to the other, and at least three have codiverged during speciation of their hosts. The results show that a variety of transfer mechanisms of Wolbachia are possible even within a single host genus. Codivergence of Wolbachia and their hosts is uncommon and provides a rare opportunity to investigate long-term Wolbachia evolution within a host lineage. Using synonymous divergence among codiverging infections and host nuclear genes, we estimate Wolbachia mutation rates to be approximately one-third that of the nuclear genome.}, } @article {pmid18820434, year = {2008}, author = {Barrett, A and Krueger, S and Jackson, M and Datta, S}, title = {Curable genetic instability of a gal4 transgene in Drosophila.}, journal = {Fly}, volume = {2}, number = {2}, pages = {67-73}, doi = {10.4161/fly.6182}, pmid = {18820434}, issn = {1933-6942}, mesh = {Animals ; Anti-Bacterial Agents ; Brain/metabolism ; Drosophila/*genetics/metabolism/microbiology ; Drosophila Proteins/*genetics ; Female ; Gene Expression ; *Gene Silencing ; Genes, Reporter ; Host-Pathogen Interactions ; Male ; Polymerase Chain Reaction ; Tetracycline ; Transcription Factors/*genetics ; *Transgenes ; }, abstract = {Transgenes, especially those driving production of the GAL4 transcription factor in a specific spatial pattern, are a critical and widely used tool in the Drosophila research community. We recently noticed loss of GAL4-driven reporter gene expression in a series of crosses, and traced that loss of reporter gene expression to stochastic physical loss of the GAL4 gene in the driver line. We have demonstrated that the instability of the GAL4 transgene can be "cured" by treatment of the line with tetracycline, suggesting that the causative agent is of bacterial origin. A PCR assay revealed that the line is not infected by Wolbachia, an intracellular parasite known to infect a large percentage of stocks in the public stock centers and to affect mutant phenotypes. Our data indicate that other tetracycline-sensitive agents can cause genetic instability of transgenes, and also provides a potential solution to the problem.}, } @article {pmid18818770, year = {2008}, author = {Braquart-Varnier, C and Lachat, M and Herbinière, J and Johnson, M and Caubet, Y and Bouchon, D and Sicard, M}, title = {Wolbachia mediate variation of host immunocompetence.}, journal = {PloS one}, volume = {3}, number = {9}, pages = {e3286}, pmid = {18818770}, issn = {1932-6203}, mesh = {Animals ; Antibody Formation ; Arthropods/*microbiology/physiology ; Cell Lineage ; DNA, Mitochondrial/metabolism ; Female ; Hemocytes/metabolism ; Hemolymph/metabolism ; Immune System ; Immunocompetence/*genetics ; Models, Biological ; Phenotype ; Symbiosis/genetics ; Wolbachia/*genetics/*physiology ; }, abstract = {BACKGROUND: After decades during which endosymbionts were considered as silent in their hosts, in particular concerning the immune system, recent studies have revealed the contrary. In the present paper, we addressed the effect of Wolbachia, the most prevalent endosymbiont in arthropods, on host immunocompetence. To this end, we chose the A. vulgare-Wolbachia symbiosis as a model system because it leads to compare consequences of two Wolbachia strains (wVulC and wVulM) on hosts from the same population. Moreover, A. vulgare is the only host-species in which Wolbachia have been directly observed within haemocytes which are responsible for both humoral and cellular immune responses.

We sampled gravid females from the same population that were either asymbiotic, infected with wVulC, or infected with wVulM. The offspring from these females were tested and it was revealed that individuals harbouring wVulC exhibited: (i) lower haemocyte densities, (ii) more intense septicaemia in their haemolymph and (iii) a reduced lifespan as compared to individuals habouring wVulM or asymbiotic ones. Therefore, individuals in this population of A. vulgare appeared to suffer more from wVulC than from wVulM. Symbiotic titer and location in the haemocytes did not differ for the two Wolbachia strains showing that these two parameters were not responsible for differences observed in their extended phenotypes in A. vulgare.

CONCLUSION/SIGNIFICANCE: The two Wolbachia strains infecting A. vulgare in the same population induced variation in immunocompetence and survival of their hosts. Such variation should highly influence the dynamics of this host-symbiont system. We propose in accordance with previous population genetic works, that wVulM is a local strain that has attenuated its virulence through a long term adaptation process towards local A. vulgare genotypes whereas wVulC, which is a widespread and invasive strain, is not locally adapted.}, } @article {pmid18814705, year = {2008}, author = {Bain, O and Casiraghi, M and Martin, C and Uni, S}, title = {The nematoda Filarioidea: critical analysis linking molecular and traditional approaches.}, journal = {Parasite (Paris, France)}, volume = {15}, number = {3}, pages = {342-348}, doi = {10.1051/parasite/2008153342}, pmid = {18814705}, issn = {1252-607X}, mesh = {Animals ; *Evolution, Molecular ; Filarioidea/*classification/genetics/*microbiology ; Genetic Markers ; Host-Pathogen Interactions ; *Phylogeny ; Symbiosis ; Wolbachia/growth & development/*pathogenicity ; }, abstract = {The molecular analysis of the Filarioidea and the endobacteria Wolbachia is no more limited to the agents of human diseases and the diversified sampling permits a synthesis with the morphological and biological results. The validity of the genera with "uncoherent host range", such as Monsonella, Litomosoides and Cercopithifilaria, is confirmed and, consequently, their evolution by host-switchings. Dirofilaria and Onchocerca, types of two subfamilies, appear more closely related than with other onchocercids. Waltonellinae from anurans and Oswaldofilariinae from reptiles have a basal position. These filariae, and some others also considered primitive, do not harbour Wolbachia. Evidence for transversal transmission of the bacteria and a second acquisition event is given with the supergroup F, identified in Monsonella, in one of the Cercopithifilaria species and in arthropods.}, } @article {pmid18801163, year = {2008}, author = {Foster, JM and Kumar, S and Ford, L and Johnston, KL and Ben, R and Graeff-Teixeira, C and Taylor, MJ}, title = {Absence of Wolbachia endobacteria in the non-filariid nematodes Angiostrongylus cantonensis and A. costaricensis.}, journal = {Parasites & vectors}, volume = {1}, number = {1}, pages = {31}, pmid = {18801163}, issn = {1756-3305}, abstract = {The majority of filarial nematodes harbour Wolbachia endobacteria, including the major pathogenic species in humans, Onchocerca volvulus, Brugia malayi and Wuchereria bancrofti. These obligate endosymbionts have never been demonstrated unequivocally in any non-filariid nematode. However, a recent report described the detection by PCR of Wolbachia in the metastrongylid nematode, Angiostrongylus cantonensis (rat lungworm), a leading cause of eosinophilic meningitis in humans. To address the intriguing possibility of Wolbachia infection in nematode species distinct from the Family Onchocercidae, we used both PCR and immunohistochemistry to screen samples of A. cantonensis and A. costaricensis for the presence of this endosymbiont. We were unable to detect Wolbachia in either species using these methodologies. In addition, bioinformatic and phylogenetic analyses of the Wolbachia gene sequences reported previously from A. cantonensis indicate that they most likely result from contamination with DNA from arthropods and filarial nematodes. This study demonstrates the need for caution in relying solely on PCR for identification of new endosymbiont strains from invertebrate DNA samples.}, } @article {pmid18797835, year = {2009}, author = {Yoshizawa, J and Mimori, K and Yamauchi, K and Tsuchida, K}, title = {Sex mosaics in a male dimorphic ant Cardiocondyla kagutsuchi.}, journal = {Die Naturwissenschaften}, volume = {96}, number = {1}, pages = {49-55}, pmid = {18797835}, issn = {0028-1042}, mesh = {Animals ; Ants/anatomy & histology/embryology/*genetics ; Chromosome Aberrations ; Female ; Larva/anatomy & histology ; Male ; *Mosaicism ; *Sex Characteristics ; }, abstract = {Gynandromorphy, or the development of organisms with a combination of male and female morphological features, is common in Hymenoptera. The underlying mechanism is likely associated with the sex-determination system, and studying this phenomenon should lead to a deeper understanding of both embryonic development and sex determination. The reproductive capabilities of gynandromorphs (hereafter, sex mosaics) remain unclear. We studied gynandromorphy in the Malaysian ant Cardiocondyla kagutsuchi, which has sex mosaics of queens (gynandromorphs; mosaic of queens and winged male) and workers (ergatandromorphs; mosaic of worker and wingless ergatoid male). These sex mosaics were classified into seven morphological categories. Most individuals had more male than female body areas. Behavioral observations revealed that sex mosaics behave more in accordance with the "sex" of their brain than that of the reproductive organs (gaster). Relative DNA quantities showed that both female and male regions contained haploid and diploid nuclei, irrespective of their phenotypic appearance, indicating that external appearance did not reflect internal tissues. Nearly one third of the adults were sex mosaics and they were not infected with Wolbachia. Our results suggest that the production of sex mosaics in this species does not pose a substantial cost to colonies and that the underlying causes are therefore not strongly selected against.}, } @article {pmid18794912, year = {2008}, author = {Werren, JH and Baldo, L and Clark, ME}, title = {Wolbachia: master manipulators of invertebrate biology.}, journal = {Nature reviews. Microbiology}, volume = {6}, number = {10}, pages = {741-751}, doi = {10.1038/nrmicro1969}, pmid = {18794912}, issn = {1740-1534}, mesh = {Animals ; Arthropods/microbiology/*physiology ; Bacterial Proteins/genetics ; Host-Parasite Interactions/genetics ; Nematoda/*microbiology/*physiology ; Symbiosis/genetics/*physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These alphaproteobacteria endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-egg incompatibility. They can also move horizontally across species boundaries, resulting in a widespread and global distribution in diverse invertebrate hosts. Here, we review the basic biology of Wolbachia, with emphasis on recent advances in our understanding of these fascinating endosymbionts.}, } @article {pmid18780877, year = {2008}, author = {Nunes, MD and Nolte, V and Schlötterer, C}, title = {Nonrandom Wolbachia infection status of Drosophila melanogaster strains with different mtDNA haplotypes.}, journal = {Molecular biology and evolution}, volume = {25}, number = {11}, pages = {2493-2498}, pmid = {18780877}, issn = {1537-1719}, support = {P 19467/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; *DNA, Mitochondrial ; Drosophila melanogaster/genetics/*microbiology ; Female ; *Haplotypes ; *Wolbachia ; }, abstract = {Wolbachia are maternally inherited bacteria, which typically spread in the host population by inducing cytoplasmic incompatibility (CI). In Drosophila melanogaster, Wolbachia is quite common but CI is variable, with most of the studies reporting low levels of CI. Surveying mitochondrial DNA (mtDNA) variation and infection status in a worldwide D. melanogaster collection, we found that the Wolbachia infection was not randomly distributed among flies with different mtDNA haplotypes. This preferential infection of some mtDNA haplotypes could be caused by a recent spread of mtDNA haplotypes associated with the infection. The comparison of contemporary D. melanogaster samples with lines collected more than 50 years ago shows that indeed one haplotype with a high incidence of Wolbachia infection has increased in frequency. Consistent with this observation, we found that the acquisition of a Wolbachia infection in a population from Crete was accompanied with an almost complete mtDNA replacement, with the Wolbachia-associated haplotype becoming abundant. Although it is difficult to identify the evolutionary forces causing the global increase of wMel, the parallel sweep of Wolbachia and an mtDNA haplotype suggests a fitness advantage of the Wolbachia infection.}, } @article {pmid18772939, year = {2008}, author = {Graham, RI and Zahner, V and Lucarotti, CJ}, title = {An intracellular symbiont and other microbiota associated with field-collected populations of sawflies (Hymenoptera: Symphyta).}, journal = {Canadian journal of microbiology}, volume = {54}, number = {9}, pages = {758-768}, doi = {10.1139/w08-067}, pmid = {18772939}, issn = {0008-4166}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; *Bacterial Physiological Phenomena ; Bacteriophages/genetics/*isolation & purification ; Canada ; Hymenoptera/*microbiology ; Molecular Sequence Data ; Phylogeny ; Polymorphism, Restriction Fragment Length ; *Symbiosis ; Wolbachia/genetics/isolation & purification/physiology/virology ; }, abstract = {Six species of sawfly (Hymenoptera: Symphyta) from four taxonomic families (Agridae, Diprionidae, Pamphiliidae, and Tenthredinidae) were collected from locations across Canada and surveyed for their associated microbiota. Total DNA was extracted from individual insects, and polymerase chain reaction (PCR) was used to amplify the conserved 16S rRNA gene from microbiota. Denaturing gradient gel electrophoresis (DGGE) and restriction fragment length polymorphism (RFLP) were undertaken to separate bacterial clones associated with the host insect. Sequencing of the PCR-DGGE and PCR--RFLP products revealed a dominance of alpha- and gamma-Proteobacteria, with most sequences showing high similarity to bacteria previously identified from other insect species and environmental samples. Additionally, a strain of the bacterial endosymbiont Wolbachia and a Wolbachia bacteriophage were identified from the mountain ash sawfly (Pristiphora geniculata).}, } @article {pmid18770621, year = {2007}, author = {Dobson, SL}, title = {Transfection of Wolbachia pipientis into Drosophila embryos.}, journal = {Current protocols in microbiology}, volume = {Chapter 3}, number = {}, pages = {Unit 3A.4}, doi = {10.1002/9780471729259.mc03a04s05}, pmid = {18770621}, issn = {1934-8533}, mesh = {Animals ; Drosophila/embryology/*microbiology ; Embryo, Nonmammalian/*microbiology ; Female ; Male ; Microinjections/methods ; *Transfection ; Wolbachia/classification/*genetics/*pathogenicity ; }, abstract = {Wolbachia is a genus of obligate intracellular Alpha-Proteobacteria represented by the type species Wolbachia pipientis (Dumler et al., 2001). Wolbachia commonly reside within cytoplasmic vacuoles of arthropods and helminths (Werren and Windsor, 2000; Casiraghi et al., 2004); vertebrate infections have not been identified. Wolbachia are maternally transmitted from mothers to offspring though the embryonic cytoplasm. Wolbachia are able to induce a diverse range of phenotypes in their invertebrate hosts, ranging from classical mutualism to reproductive parasitism. Examples of the latter include male killing, host feminization, parthenogenesis, and cytoplasmic incompatibility (reviewed in Dobson, 2003a). Current Wolbachia research foci include examining the impacts of Wolbachia infection on host evolution, characterizing the mechanisms by which Wolbachia manipulate invertebrate hosts, and developing applied strategies that employ Wolbachia for pest and disease control. Wolbachia transfection has proven a useful technique for addressing questions within each of these research foci. This unit describes a method for Wolbachia transfection via embryonic microinjection.}, } @article {pmid18763041, year = {2008}, author = {Schütte, C and Dicke, M}, title = {Verified and potential pathogens of predatory mites (Acari: Phytoseiidae).}, journal = {Experimental & applied acarology}, volume = {46}, number = {1-4}, pages = {307-328}, pmid = {18763041}, issn = {1572-9702}, mesh = {Animals ; *Host-Pathogen Interactions ; Micrococcaceae/physiology ; Microsporidia/physiology ; Mites/*microbiology/parasitology ; Pest Control, Biological ; Rickettsiaceae/physiology ; }, abstract = {Several species of phytoseiid mites (Acari: Phytoseiidae), including species of the genera Amblyseius, Galendromus, Metaseiulus, Neoseiulus, Phytoseiulus and Typhlodromus, are currently reared for biological control of various crop pests and/or as model organisms for the study of predator-prey interactions. Pathogen-free phytoseiid mites are important to obtain high efficacy in biological pest control and to get reliable data in mite research, as pathogens may affect the performance of their host or alter their reproduction and behaviour. Potential and verified pathogens have been reported for phytoseiid mites during the past 25 years. The present review provides an overview, including potential pathogens with unknown host effects (17 reports), endosymbiotic Wolbachia (seven reports), other bacteria (including Cardinium and Spiroplasma) (four reports), cases of unidentified diseases (three reports) and cases of verified pathogens (six reports). From the latter group four reports refer to Microsporidia, one to a fungus and one to a bacterium. Only five entities have been studied in detail, including Wolbachia infecting seven predatory mite species, other endosymbiotic bacteria infecting Metaseiulus (Galendromus, Typhlodromus) occidentalis (Nesbitt), the bacterium Acaricomes phytoseiuli infecting Phytoseiulus persimilis Athias-Henriot, the microsporidium Microsporidium phytoseiuli infecting P. persimilis and the microsporidium Oligosproridium occidentalis infecting M. occidentalis. In four cases (Wolbachia, A. phytoseiuli, M. phytoseiuli and O. occidentalis) an infection may be connected with fitness costs of the host. Moreover, infection is not always readily visible as no obvious gross symptoms are present. Monitoring of these entities on a routine and continuous basis should therefore get more attention, especially in commercial mass-production. Special attention should be paid to field-collected mites before introduction into the laboratory or mass rearing, and to mites that are exchanged among rearing facilities. However, at present general pathogen monitoring is not yet practical as effects of many entities are unknown. More research effort is needed concerning verified and potential pathogens of commercially reared arthropods and those used as model organisms in research.}, } @article {pmid18758845, year = {2009}, author = {Gibson, CM and Hunter, MS}, title = {Inherited fungal and bacterial endosymbionts of a parasitic wasp and its cockroach host.}, journal = {Microbial ecology}, volume = {57}, number = {3}, pages = {542-549}, pmid = {18758845}, issn = {1432-184X}, mesh = {Animals ; Bacteria/classification/*genetics/isolation & purification ; Bayes Theorem ; *Biological Evolution ; Cockroaches/*microbiology/parasitology ; DNA, Bacterial/genetics ; DNA, Fungal/genetics ; Fungi/classification/*genetics/isolation & purification ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; *Symbiosis ; Wasps/*microbiology ; }, abstract = {Bacterial endosymbionts of insects are increasingly being recognized as common, diverse, and integral to the biology of their hosts. Inherited fungal symbionts have been largely overlooked, however, even though insect guts appear to be a key habitat for an incredible array of fungal diversity. Like bacteria, fungal symbionts also likely play important roles in the ecology and evolution of their insect associates. The objective of this study was to lay the foundations for understanding the roles of the vertically transmitted fungal and bacterial associates of both the brownbanded cockroach, Supella longipalpa, and its parasitic wasp, Comperia merceti. We used culture-dependent and culture-independent molecular methods and phylogenetic analyses in order to identify the symbionts. Two fungal associates of brownbanded cockroaches were found. To our knowledge, this is the first record of vertically transmitted fungal symbionts in the order Blattaria. The wasp was found to house a close relative of one of the cockroach fungi but no bacterial symbionts. Finally, the brownbanded cockroaches also harbored three lineages of bacterial symbionts: Blattabacterium and two lineages of Wolbachia, indicating the number of vertically transmitted symbionts in this insect may be as many as five.}, } @article {pmid18755670, year = {2008}, author = {Jansen, VA and Turelli, M and Godfray, HC}, title = {Stochastic spread of Wolbachia.}, journal = {Proceedings. Biological sciences}, volume = {275}, number = {1652}, pages = {2769-2776}, pmid = {18755670}, issn = {0962-8452}, mesh = {Animals ; Female ; *Genetics, Population ; Insecta/*microbiology ; *Models, Biological ; Population Dynamics ; *Stochastic Processes ; Wolbachia/*growth & development ; }, abstract = {Wolbachia are very common, maternally transmitted endosymbionts of insects. They often spread by a mechanism termed cytoplasmic incompatibility (CI) that involves reduced egg hatch when Wolbachia-free ova are fertilized by sperm from Wolbachia-infected males. Because the progeny of Wolbachia-infected females generally do not suffer CI-induced mortality, infected females are often at a reproductive advantage in polymorphic populations. Deterministic models show that Wolbachia that impose no costs on their hosts and have perfect maternal transmission will spread from arbitrarily low frequencies (though initially very slowly); otherwise, there will be a threshold frequency below which Wolbachia frequencies decline to extinction and above which they increase to fixation or a high stable equilibrium. Stochastic theory was used to calculate the probability of fixation in populations of different size for arbitrary current frequencies of Wolbachia, with special attention paid to the case of spread after the arrival of a single infected female. Exact results are given based on a Moran process that assumes a specific demographic model, and approximate results are obtained using the more general Wright-Fisher theory. A new analytical approximation for the probability of fixation is derived, which performs well for small population sizes. The significance of stochastic effects in the natural spread of Wolbachia and their relevance to the use of Wolbachia as a drive mechanism in vector and pest management are discussed.}, } @article {pmid18726699, year = {2008}, author = {Bjørnson, S}, title = {Natural enemies of mass-reared predatory mites (family Phytoseiidae) used for biological pest control.}, journal = {Experimental & applied acarology}, volume = {46}, number = {1-4}, pages = {299-306}, pmid = {18726699}, issn = {1572-9702}, mesh = {Animals ; *Host-Pathogen Interactions ; Microsporidia/physiology ; Mites/*microbiology ; *Pest Control, Biological ; Predatory Behavior ; }, abstract = {Predatory mites of the family Phytoseiidae are valued natural enemies that provide effective pest control in greenhouses and on agricultural crops. Mass-reared phytoseiids are occasionally associated with microorganisms and although their effects are not always apparent, some are pathogenic and reduce host fitness. Invertebrate pathogens are encountered more frequently in mass production systems than in nature because rearing environments often cause overcrowding and other stresses that favour pathogen transmission and increase an individual's susceptibility to disease. Although unidentified microorganisms have been reported in phytoseiids, bacteria and microsporidia have been detected with considerable frequency. The bacterium Acaricomes phytoseiuli is associated with an accumulation of birefringent crystals in the legs of Phytoseiulus persimilis and infection reduces the fitness of this spider mite predator. Wolbachia, detected in Metaseiulus occidentalis and other phytoseiids, may cause cytoplasmic incompatibilities that affect fecundity. However, the effects of Rickettsiella phytoseiuli on P. persimilis are unknown. Microsporidia are spore-forming pathogens that infect Neoseiulus cucumeris, N. barkeri, M. occidentalis and P. persimilis. Microsporidia cause chronic, debilitating disease and these pathogens often remain undetected in mass-rearings until a decrease in productivity is noticed. Routine screening of individuals is important to prevent diseased mites from being introduced into existing mass-rearings and to ensure that mite populations remain free from pathogens. The means by which bacteria and microsporidia are detected and strategies for their management in phytoseiid mass-rearings are discussed.}, } @article {pmid18717746, year = {2008}, author = {Jaenike, J and Dyer, KA}, title = {No resistance to male-killing Wolbachia after thousands of years of infection.}, journal = {Journal of evolutionary biology}, volume = {21}, number = {6}, pages = {1570-1577}, doi = {10.1111/j.1420-9101.2008.01607.x}, pmid = {18717746}, issn = {1420-9101}, support = {EF-0328363//PHS HHS/United States ; }, mesh = {Animals ; Drosophila/*genetics/*microbiology ; Electron Transport Complex IV/genetics ; Female ; Genetic Variation ; Host-Pathogen Interactions/physiology ; Male ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Maternally transmitted male-killing endosymbionts can exert strong and relentless selection pressure on their hosts to evolve resistance to these infections. Surveys of current infection prevalence and mtDNA diversity indicate that Drosophila innubila is and has been infected with male-killing Wolbachia at moderate frequencies for extended evolutionary periods. Here, we use coalescent simulations to infer the minimum age of the Wolbachia infection in this species, and estimate that the infection is at least 15,000 and perhaps over 700,000 years old. We also surveyed this species for genetic variation for resistance to the male-killing effects of infection. Our surveys revealed no evidence for any resistance polymorphism, such that all flies are completely susceptible to male killing. Given the general assumption that Drosophila can be selected for anything, the lack of resistance, despite thousands of years of strong selection, is an apparent evolutionary conundrum. We hypothesize that resistance requires a mutation of major effect that acts early in development, and that the adverse pleiotropic consequences of such mutations in both infected and uninfected individuals may exceed the possible benefit to infected flies.}, } @article {pmid18713719, year = {2008}, author = {Adachi-Hagimori, T and Miura, K and Stouthamer, R}, title = {A new cytogenetic mechanism for bacterial endosymbiont-induced parthenogenesis in Hymenoptera.}, journal = {Proceedings. Biological sciences}, volume = {275}, number = {1652}, pages = {2667-2673}, pmid = {18713719}, issn = {0962-8452}, mesh = {Animals ; Bacteroidetes ; Base Sequence ; Cytogenetic Analysis ; DNA Primers/genetics ; Hymenoptera/*microbiology/*physiology ; Meiosis/*physiology ; Microsatellite Repeats/genetics ; Molecular Sequence Data ; Parthenogenesis/*physiology ; Rickettsia ; Symbiosis/*physiology ; Wolbachia ; }, abstract = {Vertically transmitted endosymbiotic bacteria, such as Wolbachia, Cardinium and Rickettsia, modify host reproduction in several ways to facilitate their own spread. One such modification results in parthenogenesis induction, where males, which are unable to transmit the bacteria, are not produced. In Hymenoptera, the mechanism of diploidization due to Wolbachia infection, known as gamete duplication, is a post-meiotic modification. During gamete duplication, the meiotic mechanism is normal, but in the first mitosis the anaphase is aborted. The two haploid sets of chromosomes do not separate and thus result in a single nucleus containing two identical sets of haploid chromosomes. Here, we outline an alternative cytogenetic mechanism for bacterial endosymbiont-induced parthenogenesis in Hymenoptera. During female gamete formation in Rickettsia-infected Neochrysocharis formosa (Westwood) parasitoids, meiotic cells undergo only a single equational division followed by the expulsion of a single polar body. This absence of meiotic recombination and reduction corresponds well with a non-segregation pattern in the offspring of heterozygous females. We conclude that diploidy in N. formosa is maintained through a functionally apomictic cloning mechanism that differs entirely from the mechanism induced by Wolbachia.}, } @article {pmid18713031, year = {2008}, author = {Serbus, LR and Casper-Lindley, C and Landmann, F and Sullivan, W}, title = {The genetics and cell biology of Wolbachia-host interactions.}, journal = {Annual review of genetics}, volume = {42}, number = {}, pages = {683-707}, doi = {10.1146/annurev.genet.41.110306.130354}, pmid = {18713031}, issn = {0066-4197}, support = {GM 080192-02/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/genetics/microbiology ; Female ; Host-Pathogen Interactions/*genetics/*physiology ; Insecta/*genetics/*microbiology ; Male ; Oocytes/microbiology ; Oogenesis ; Spermatogenesis ; Spermatozoa/microbiology ; Wolbachia/*genetics/pathogenicity/*physiology ; }, abstract = {Wolbachia are gram-negative bacteria that are widespread in nature, carried by the majority of insect species as well as some mites, crustaceans, and filarial nematodes. Wolbachia can range from parasitic to symbiotic, depending upon the interaction with the host species. The success of Wolbachia is attributed to efficient maternal transmission and manipulations of host reproduction that favor infected females, such as sperm-egg cytoplasmic incompatibility (CI). Much remains unknown about the mechanistic basis for Wolbachia-host interactions. Here we summarize the current understanding of Wolbachia interaction with insect hosts, with a focus on Drosophila. The areas of discussion include Wolbachia transmission in oogenesis, Wolbachia distribution in spermatogenesis, induction and rescue of the CI phenotype, Wolbachia genomics, and Wolbachia-membrane interactions.}, } @article {pmid18707453, year = {2002}, author = {Telschow, A and Hammerstein, P and Werren, JH}, title = {The effect of Wolbachia on genetic divergence between populations: models with two-way migration.}, journal = {The American naturalist}, volume = {160 Suppl 4}, number = {}, pages = {S54-66}, doi = {10.1086/342153}, pmid = {18707453}, issn = {1537-5323}, abstract = {Wolbachia are intracellular bacteria that cause various reproduction alterations in their hosts, including cytoplasmic incompatibility (CI), an incompatibility between sperm and egg that typically results in embryonic death. We investigate theoretically the effects of Wolbachia-induced bidirectional CI on levels of divergence between two populations, where there is migration in both directions and differential selection at a single locus. The main findings are as follows: Wolbachia differences in the two populations are maintained up to a threshold migration rate, above which the system collapses to a single Wolbachia type; differential selection at a nuclear locus increases the threshold migration rate below which Wolbachia polymorphisms are maintained; Wolbachia differences between the populations enhance their genetic divergence at the selected locus by reducing the "effective migration rate," and even moderate levels of CI can cause large population differences in allele frequencies; and asymmetric CI can induce strong asymmetries in effective migration rate and dramatically alter the pattern of genetic divergence compared with the No Wolbachia situation. We derive an analytical approximation for the effective migration rate, which matches the simulation results for most parameter values. These results generally support the view that CI Wolbachia can contribute to genetic divergence between populations.}, } @article {pmid18707445, year = {2002}, author = {Hurst, GD and Jiggins, FM and Pomiankowski, A}, title = {Which way to manipulate host reproduction? Wolbachia that cause cytoplasmic incompatibility are easily invaded by sex ratio-distorting mutants.}, journal = {The American naturalist}, volume = {160}, number = {3}, pages = {360-373}, doi = {10.1086/341524}, pmid = {18707445}, issn = {1537-5323}, abstract = {The bacterium Wolbachia manipulates its hosts by inducing cytoplasmic incompatibility (CI), where zygotes formed from crosses between uninfected mothers and infected fathers die. In addition, it distorts the host's sex ratio via male killing, parthenogenesis induction, or feminization. Here, we model transitions between these states, examining the evolution of mutants of CI strains that retain both the ability to induce and resist CI but, in addition, cause sex ratio distortion. The model shows that CI strains are highly susceptible to invasion and subsequent elimination by these mutants. For all three types of sex ratio distortion, there is some parameter space in which the strain showing sex ratio distortion becomes extinct following exclusion of the progenitor CI strain, leaving the population uninfected. Extinction of the new Wolbachia strain is common for the case of male killing but rarer for parthenogenesis induction and feminization. Our models predict that CI strains of Wolbachia will occur most commonly in hosts that are male heterogametic, where there is little interaction between siblings because these hosts are unlikely to favor the spread of male killing, feminization, or parthenogenesis induction. The models raise the question of why CI strains apparently predominate in nature, and it is suggested that this is a result of either fewer restrictions on CI strains spreading through novel host populations or restrictions to the mutability of Wolbachia strains.}, } @article {pmid18697321, year = {2008}, author = {Ahantarig, A and Trinachartvanit, W and Kittayapong, P}, title = {Relative Wolbachia density of field-collected Aedes albopictus mosquitoes in Thailand.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {33}, number = {1}, pages = {173-177}, doi = {10.3376/1081-1710(2008)33[173:rwdofa]2.0.co;2}, pmid = {18697321}, issn = {1081-1710}, mesh = {Aedes/*microbiology ; Animals ; Female ; Host-Pathogen Interactions ; Polymerase Chain Reaction ; Thailand ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Female Aedes albopictus mosquitoes from natural populations of different geographical regions of Thailand were collected and allowed to oviposit to determine relative Wolbachia A and Wolbachia B densities of their offspring (F1) by using real-time quantitative PCR (RTQ-PCR). An important aspect of this work is that all Aedes albopictus mosquitoes were collected from the field. Twenty-seven offspring were from diverse areas of Thailand (Songkhla, Konkaen, Chantaburi, and Kanchanaburi). The range of relative Wolbachia A density in F, mosquitoes was from 0.007 to 1250.78 (bacteria-to-host ratio), whereas relative Wolbachia B densities ranged from 0 to 348.2 (bacteria-to-host ratio). These data are in contrast to those from a previous study that showed a very low amount (less than 0.10) of both relative Wolbachia density types for laboratory strains. The percent transmission of Wolbachia density from mother to each individual offspring cannot be predicted and was not related to the sex of the F1. Obtaining confirmation for variations and unpredictable Wolbachia transmission load raises some concerns about using Wolbachia as a gene-driving system in nature for population replacement if Wolbachia density is involved in cytoplasmic incompatibility in this mosquito.}, } @article {pmid18696231, year = {2008}, author = {Hoy, MA and Jeyaprakash, A}, title = {Symbionts, including pathogens, of the predatory mite Metaseiulus occidentalis: current and future analysis methods.}, journal = {Experimental & applied acarology}, volume = {46}, number = {1-4}, pages = {329-347}, pmid = {18696231}, issn = {1572-9702}, mesh = {Animals ; *Host-Pathogen Interactions ; Microsporidia/physiology ; Mites/genetics/*microbiology ; Mitochondria/genetics ; Polymerase Chain Reaction ; Rickettsia/physiology ; Serratia marcescens/physiology ; *Symbiosis ; Wolbachia/physiology ; }, abstract = {Metaseiulus (= Typhlodromus or Galendromus) occidentalis (Nesbitt) (Acari: Phytoseiidae) is an effective natural enemy of pest mites in a variety of crops around the world, although it is considered to be endemic in the western USA. A broad understanding of much of its biology, ecology, behavior, and genetics has been obtained over the past 60 years, but the role(s) symbionts play, which includes pathogens and other microorganisms, remains to be resolved fully. Until molecular tools became available, analysis methods were limited primarily to microscopic observations; some viruses and rickettsia-like organisms were observed infecting 'diseased' M. occidentalis, but it is not clear which one(s) was the causal agent(s) of the disease(s). Subsequent to the development of the polymerase chain reaction (PCR) and genome sequencing, we identified putative gut symbionts and reproductive tract symbionts in M. occidentalis, as well as a microsporidian pathogen. A new phylogenetic analysis of the Bacteroidetes-Flavobacterium group suggests the unnamed Bacteroidetes in M. occidentalis is associated with the digestive tract. However, much of our current information about the role these microorganisms play in the biology of M. occidentalis is based on correlation, lacking the strength of fulfilling Koch's postulates. We also currently lack any knowledge of the importance of these microorganisms under field conditions. In the future, it should be possible to learn what role(s) these organisms play in the biology of this important predator using metagenomics approaches to analyze the transcriptome and to determine their relative abundance within their hosts with the quantitative PCR. We have just begun to resolve these relationships.}, } @article {pmid18685571, year = {2008}, author = {Maroja, LS and Clark, ME and Harrison, RG}, title = {Wolbachia plays no role in the one-way reproductive incompatibility between the hybridizing field crickets Gryllus firmus and G. pennsylvanicus.}, journal = {Heredity}, volume = {101}, number = {5}, pages = {435-444}, doi = {10.1038/hdy.2008.75}, pmid = {18685571}, issn = {1365-2540}, mesh = {Animals ; Crosses, Genetic ; Female ; Gryllidae/genetics/*microbiology/*physiology ; *Hybridization, Genetic ; Male ; Ovum/cytology/microbiology ; Polymorphism, Restriction Fragment Length ; Reproduction ; Sexual Behavior, Animal ; Species Specificity ; Spermatozoa/cytology/microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are cytoplasmically inherited alpha-proteobacteria that can cause cytoplasmic incompatibility (CI) in insects. This incompatibility between sperm and egg is evident when uninfected females mate with infected males. Wolbachia-driven reproductive incompatibilities are of special interest because they may play a role in speciation. However, the presence of Wolbachia does not always imply incompatibility. The field crickets Gryllus firmus and G. pennsylvanicus exhibit a very clear unidirectional incompatibility and have been cited as a possible example of Wolbachia-induced CI. Here, we conduct curing experiments, intra- and interspecific crosses, cytological examination of Wolbachia in testes and Wolbachia quantifications through real-time PCR. All of our data strongly suggest that Wolbachia are not involved in the reproductive incompatibility between G. firmus and G. pennsylvanicus.}, } @article {pmid18679718, year = {2008}, author = {Specht, S and Mand, S and Marfo-Debrekyei, Y and Debrah, AY and Konadu, P and Adjei, O and Büttner, DW and Hoerauf, A}, title = {Efficacy of 2- and 4-week rifampicin treatment on the Wolbachia of Onchocerca volvulus.}, journal = {Parasitology research}, volume = {103}, number = {6}, pages = {1303-1309}, pmid = {18679718}, issn = {0932-0113}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/administration & dosage/pharmacology/*therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Onchocerca volvulus/drug effects/embryology/growth & development/*microbiology ; Onchocerciasis/*drug therapy/parasitology ; Rifampin/administration & dosage/pharmacology/*therapeutic use ; Treatment Outcome ; Wolbachia/classification/*drug effects ; Young Adult ; }, abstract = {The microfilaricidal and temporarily sterilizing drug ivermectin is used for mass treatment of filarial infections. Filariae containing Wolbachia endobacteria can also be treated by the antibiotic doxycycline. The loss of Wolbachia results in sterilization of Onchocerca volvulus and macrofilaricidal effects. Besides doxycycline, other antibiotics may be effective in depleting Wolbachia. A preliminary study on the effects of rifampicin on the endobacteria, embryogenesis and microfilariae production of O. volvulus was carried out in the year 2000 in Ghana. Twenty-six onchocerciasis patients were treated for 2 or 4 weeks with 10 mg/kg/day rifampicin. From 17 treated and nine untreated patients, all palpable nodules were extirpated 1 or 18 months after the start of the study and examined for Wolbachia and embryogenesis using immunohistology. One and 18 months after rifampicin treatment, the proportion of Wolbachia-positive worms was significantly reduced compared to the untreated group. In patients treated 4 weeks with rifampicin, only 21% and 18% of living female filariae contained Wolbachia after 1 and 18 months, respectively, compared to 92% in the untreated patients. The reduction of Wolbachia after 2 weeks rifampicin was less but also significant. Embryogenesis and microfilariae production were reduced after 4 weeks rifampicin treatment, rendering rifampicin an antibiotic with anti-wolbachial efficacy in human onchocerciasis. This treatment is less efficient than treatment with 6 weeks doxycycline, but might be an alternative for cases that cannot be treated with doxycycline, e.g. children, or might be further developed for combination therapy.}, } @article {pmid18653734, year = {2008}, author = {Oliveira, DC and Raychoudhury, R and Lavrov, DV and Werren, JH}, title = {Rapidly evolving mitochondrial genome and directional selection in mitochondrial genes in the parasitic wasp nasonia (hymenoptera: pteromalidae).}, journal = {Molecular biology and evolution}, volume = {25}, number = {10}, pages = {2167-2180}, pmid = {18653734}, issn = {1537-1719}, support = {R01 GM070026/GM/NIGMS NIH HHS/United States ; 5R01 GM070026/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cell Nucleus/metabolism ; DNA, Mitochondrial/*genetics ; Evolution, Molecular ; Expressed Sequence Tags ; Genetic Variation ; Genome ; *Genome, Mitochondrial ; Hymenoptera/*genetics/parasitology ; Mitochondria/metabolism ; Models, Genetic ; *Mutation ; Phylogeny ; Wasps/genetics ; Wolbachia/*genetics ; }, abstract = {We sequenced the nearly complete mtDNA of 3 species of parasitic wasps, Nasonia vitripennis (2 strains), Nasonia giraulti, and Nasonia longicornis, including all 13 protein-coding genes and the 2 rRNAs, and found unusual patterns of mitochondrial evolution. The Nasonia mtDNA has a unique gene order compared with other insect mtDNAs due to multiple rearrangements. The mtDNAs of these wasps also show nucleotide substitution rates over 30 times faster than nuclear protein-coding genes, indicating among the highest substitution rates found in animal mitochondria (normally <10 times faster). A McDonald and Kreitman test shows that the between-species frequency of fixed replacement sites relative to silent sites is significantly higher compared with within-species polymorphisms in 2 mitochondrial genes of Nasonia, atp6 and atp8, indicating directional selection. Consistent with this interpretation, the Ka/Ks (nonsynonymous/synonymous substitution rates) ratios are higher between species than within species. In contrast, cox1 shows a signature of purifying selection for amino acid sequence conservation, although rates of amino acid substitutions are still higher than for comparable insects. The mitochondrial-encoded polypeptides atp6 and atp8 both occur in F0F1ATP synthase of the electron transport chain. Because malfunction in this fundamental protein severely affects fitness, we suggest that the accelerated accumulation of replacements is due to beneficial mutations necessary to compensate mild-deleterious mutations fixed by random genetic drift or Wolbachia sweeps in the fast evolving mitochondria of Nasonia. We further propose that relatively high rates of amino acid substitution in some mitochondrial genes can be driven by a "Compensation-Draft Feedback"; increased fixation of mildly deleterious mutations results in selection for compensatory mutations, which lead to fixation of additional deleterious mutations in nonrecombining mitochondrial genomes, thus accelerating the process of amino acid substitutions.}, } @article {pmid18648384, year = {2008}, author = {Clark, ME and Bailey-Jourdain, C and Ferree, PM and England, SJ and Sullivan, W and Windsor, DM and Werren, JH}, title = {Wolbachia modification of sperm does not always require residence within developing sperm.}, journal = {Heredity}, volume = {101}, number = {5}, pages = {420-428}, doi = {10.1038/hdy.2008.71}, pmid = {18648384}, issn = {1365-2540}, mesh = {Animals ; Coleoptera/growth & development/microbiology ; Female ; Male ; Species Specificity ; *Spermatogenesis ; Spermatozoa/*growth & development/*microbiology ; Wasps/*growth & development/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacteria known to manipulate the reproduction of their arthropod hosts. Wolbachia commonly affect the sperm of infected arthropods. Wolbachia-modified sperm cannot successfully fertilize unless the female is infected with the same Wolbachia type. A study of spermatogenesis in the parasitic wasp Nasonia vitripennis reveals that Wolbachia are not required in individual spermatocytes or spermatids to modify sperm. In N. vitripennis, Wolbachia modify nearly all sperm, but are found only in approximately 28% of developing sperm, and are also found in surrounding cyst and sheath cells. In the beetle Chelymorpha alternans, Wolbachia can modify up to 90% of sperm, but were never observed within the developing sperm or within the surrounding cyst cells; they were abundant within the outer testis sheath. We conclude that the residence within a developing sperm is not a prerequisite for Wolbachia-induced sperm modification, suggesting that Wolbachia modification of sperm may occur across multiple tissue membranes or act upstream of spermiogenesis.}, } @article {pmid18622662, year = {2008}, author = {Fallon, AM and Hellestad, VJ}, title = {Standardization of a colorimetric method to quantify growth and metabolic activity of Wolbachia-infected mosquito cells.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {44}, number = {8-9}, pages = {351-356}, pmid = {18622662}, issn = {1071-2690}, support = {R21 AI070913/AI/NIAID NIH HHS/United States ; R21 AI070913-02/AI/NIAID NIH HHS/United States ; AI 070913/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cells, Cultured ; Colorimetry/methods/*standards ; Culicidae/*metabolism/*microbiology ; Mitochondria/metabolism ; Reference Standards ; Tetracycline/pharmacology ; Time Factors ; *Wolbachia/drug effects ; }, abstract = {The Aedes albopictus Aa23 cell line, which is persistently infected with Wolbachia pipientis strain wAlbB, tends to grow as aggregated clusters of cells that are difficult to disperse for conventional quantification based on cell number. We used A. albopictus C7-10 cells to validate conversion of methylthiazole tetrazolium (MTT) to a colored formazan product with respect to incubation time, cell number over a 40-fold range, and metabolic activity as cells enter stationary phase. Using this assay, we showed that the doubling time of Aa23 cells increases from about 45 h early after plating to more than 70 h as the cells reach stationary levels. Growth of Aa23 cells proceeds at similar rates in the presence or absence of tetracycline concentrations that decrease the abundance of Wolbachia. Insofar as the MTT assay reflects mitochondrial function, our results indicate that, in Aa23 cells, abundance of intracellular Wolbachia has no measurable effect on mitochondrial activity in the presence of tetracycline.}, } @article {pmid18582562, year = {2008}, author = {Félix, C and Pichon, S and Braquart-Varnier, C and Braig, H and Chen, L and Garrett, RA and Martin, G and Grève, P}, title = {Characterization and transcriptional analysis of two gene clusters for type IV secretion machinery in Wolbachia of Armadillidium vulgare.}, journal = {Research in microbiology}, volume = {159}, number = {6}, pages = {481-485}, doi = {10.1016/j.resmic.2008.05.007}, pmid = {18582562}, issn = {0923-2508}, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Biological Transport ; Female ; Feminization ; Isopoda/*microbiology ; Male ; Molecular Sequence Data ; *Multigene Family ; *Transcription, Genetic ; Wolbachia/*genetics/metabolism ; }, abstract = {Wolbachia are maternally inherited alpha-proteobacteria that induce feminization of genetic males in most terrestrial crustacean isopods. Two clusters of vir genes for a type IV secretion machinery have been identified at two separate loci and characterized for the first time in a feminizing Wolbachia. Furthermore, we demonstrated that these operons are transcriptionally active in ovaries and in all other tissues tested, suggesting that T4SS has a significant role in Wolbachia biology. These observations and the identification of homologous vir genes in Wolbachia strains infecting insects or nematodes show that vir genes are conserved among Wolbachia strains whatever the phenotype induced by the bacteria.}, } @article {pmid18577218, year = {2008}, author = {Duron, O and Bouchon, D and Boutin, S and Bellamy, L and Zhou, L and Engelstädter, J and Hurst, GD}, title = {The diversity of reproductive parasites among arthropods: Wolbachia do not walk alone.}, journal = {BMC biology}, volume = {6}, number = {}, pages = {27}, pmid = {18577218}, issn = {1741-7007}, mesh = {Animals ; Arthropods/*microbiology ; Bacteria/classification/genetics/*isolation & purification ; Bacterial Physiological Phenomena ; *Biodiversity ; Female ; Male ; Molecular Sequence Data ; Phylogeny ; Wolbachia/*growth & development/*physiology ; }, abstract = {BACKGROUND: Inherited bacteria have come to be recognised as important components of arthropod biology. In addition to mutualistic symbioses, a range of other inherited bacteria are known to act either as reproductive parasites or as secondary symbionts. Whilst the incidence of the alpha-proteobacterium Wolbachia is relatively well established, the current knowledge of other inherited bacteria is much weaker. Here, we tested 136 arthropod species for a range of inherited bacteria known to demonstrate reproductive parasitism, sampling each species more intensively than in past surveys.

RESULTS: The inclusion of inherited bacteria other than Wolbachia increased the number of infections recorded in our sample from 33 to 57, and the proportion of species infected from 22.8% to 32.4%. Thus, whilst Wolbachia remained the dominant inherited bacterium, it alone was responsible for around half of all inherited infections of the bacteria sampled, with members of the Cardinium, Arsenophonus and Spiroplasma ixodetis clades each occurring in 4% to 7% of all species. The observation that infection was sometimes rare within host populations, and that there was variation in presence of symbionts between populations indicates that our survey will itself underscore incidence.

CONCLUSION: This extensive survey demonstrates that at least a third of arthropod species are infected by a diverse assemblage of maternally inherited bacteria that are likely to strongly influence their hosts' biology, and indicates an urgent need to establish the nature of the interaction between non-Wolbachia bacteria and their hosts.}, } @article {pmid18562339, year = {2008}, author = {Cordaux, R and Pichon, S and Ling, A and Pérez, P and Delaunay, C and Vavre, F and Bouchon, D and Grève, P}, title = {Intense transpositional activity of insertion sequences in an ancient obligate endosymbiont.}, journal = {Molecular biology and evolution}, volume = {25}, number = {9}, pages = {1889-1896}, pmid = {18562339}, issn = {1537-1719}, mesh = {Animals ; Crustacea/microbiology ; *DNA Transposable Elements ; *DNA, Bacterial ; *Evolution, Molecular ; Gene Transfer, Horizontal ; Insecta/microbiology ; Molecular Sequence Data ; Sequence Homology, Nucleic Acid ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {The streamlined genomes of ancient obligate endosymbionts generally lack transposable elements, such as insertion sequences (IS). Yet, the genome of Wolbachia, one of the most abundant bacterial endosymbionts on Earth, is littered with IS. Such a paradox raises the question as to why there are so many ISs in the genome of this ancient endosymbiont. To address this question, we investigated IS transpositional activity in the unculturable Wolbachia by tracking the evolutionary dynamics and history of ISWpi1 elements. We show that 1) ISWpi1 is widespread in Wolbachia, being present in at least 55% of the 40 sampled strains, 2) ISWpi1 copies exhibit virtually identical nucleotide sequences both within and among Wolbachia genomes and possess an intact transposase gene, 3) individual ISWpi1 copies are differentially inserted among Wolbachia genomes, and 4) ISWpi1 occurs at variable copy numbers among Wolbachia genomes. Collectively, our results provide compelling evidence for intense ISWpi1 transpositional activity and frequent ISWpi1 horizontal transmission among strains during recent Wolbachia evolution. Thus, the genomes of ancient obligate endosymbionts can carry high loads of functional and transpositionally active transposable elements. Our results also indicate that Wolbachia genomes have experienced multiple and temporally distinct ISWpi1 invasions during their evolutionary history. Such recurrent exposition to new IS invasions may explain, at least partly, the unusually high density of transposable elements found in the genomes of Wolbachia endosymbionts.}, } @article {pmid18550617, year = {2008}, author = {Klasson, L and Walker, T and Sebaihia, M and Sanders, MJ and Quail, MA and Lord, A and Sanders, S and Earl, J and O'Neill, SL and Thomson, N and Sinkins, SP and Parkhill, J}, title = {Genome evolution of Wolbachia strain wPip from the Culex pipiens group.}, journal = {Molecular biology and evolution}, volume = {25}, number = {9}, pages = {1877-1887}, pmid = {18550617}, issn = {1537-1719}, support = {/WT_/Wellcome Trust/United Kingdom ; 079059/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Ankyrin Repeat/genetics ; Bacterial Proteins/genetics/metabolism ; Culex/*microbiology ; DNA, Bacterial ; Drosophila melanogaster/microbiology ; *Evolution, Molecular ; Gene Duplication ; Gene Transfer, Horizontal ; *Genome, Bacterial ; Molecular Sequence Data ; Prophages/genetics ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {The obligate intracellular bacterium Wolbachia pipientis strain wPip induces cytoplasmic incompatibility (CI), patterns of crossing sterility, in the Culex pipiens group of mosquitoes. The complete sequence is presented of the 1.48-Mbp genome of wPip which encodes 1386 coding sequences (CDSs), representing the first genome sequence of a B-supergroup Wolbachia. Comparisons were made with the smaller genomes of Wolbachia strains wMel of Drosophila melanogaster, an A-supergroup Wolbachia that is also a CI inducer, and wBm, a mutualist of Brugia malayi nematodes that belongs to the D-supergroup of Wolbachia. Despite extensive gene order rearrangement, a core set of Wolbachia genes shared between the 3 genomes can be identified and contrasts with a flexible gene pool where rapid evolution has taken place. There are much more extensive prophage and ankyrin repeat encoding (ANK) gene components of the wPip genome compared with wMel and wBm, and both are likely to be of considerable importance in wPip biology. Five WO-B-like prophage regions are present and contain some genes that are identical or highly similar in multiple prophage copies, whereas other genes are unique, and it is likely that extensive recombination, duplication, and insertion have occurred between copies. A much larger number of genes encode ankyrin repeat (ANK) proteins in wPip, with 60 present compared with 23 in wMel, many of which are within or close to the prophage regions. It is likely that this pattern is partly a result of expansions in the wPip lineage, due for example to gene duplication, but their presence is in some cases more ancient. The wPip genome underlines the considerable evolutionary flexibility of Wolbachia, providing clear evidence for the rapid evolution of ANK-encoding genes and of prophage regions. This host-Wolbachia system, with its complex patterns of sterility induced between populations, now provides an excellent model for unraveling the molecular systems underlying host reproductive manipulation.}, } @article {pmid18545272, year = {2008}, author = {Kounatidis, I and Papadopoulos, N and Bourtzis, K and Mavragani-Tsipidou, P}, title = {Genetic and cytogenetic analysis of the fruit fly Rhagoletis cerasi (Diptera: Tephritidae).}, journal = {Genome}, volume = {51}, number = {7}, pages = {479-491}, doi = {10.1139/G08-032}, pmid = {18545272}, issn = {0831-2796}, mesh = {Animals ; Chromosomes/ultrastructure ; Karyotyping ; Male ; Mitosis/genetics ; Tephritidae/*genetics/microbiology/ultrastructure ; Wolbachia/isolation & purification ; }, abstract = {The European cherry fruit fly, Rhagoletis cerasi, is a major agricultural pest for which biological, genetic, and cytogenetic information is limited. We report here a cytogenetic analysis of 4 natural Greek populations of R. cerasi, all of them infected with the endosymbiotic bacterium Wolbachia pipientis. The mitotic karyotype and detailed photographic maps of the salivary gland polytene chromosomes of this pest species are presented here. The mitotic metaphase complement consists of 6 pairs of chromosomes, including one pair of heteromorphic sex chromosomes, with the male being the heterogametic sex. The analysis of the salivary gland polytene complement has shown a total of 5 long chromosomes (10 polytene arms) that correspond to the 5 autosomes of the mitotic nuclei and a heterochromatic mass corresponding to the sex chromosomes. The most prominent landmarks of each polytene chromosome, the "weak points", and the unusual asynapsis of homologous pairs of polytene chromosomes at certain regions of the polytene elements are also presented and discussed.}, } @article {pmid18541758, year = {2008}, author = {Mand, S and Büttner, DW and Hoerauf, A}, title = {Bancroftian filariasis--absence of Wolbachia after doxycycline treatment.}, journal = {The American journal of tropical medicine and hygiene}, volume = {78}, number = {6}, pages = {854-855}, pmid = {18541758}, issn = {1476-1645}, mesh = {Doxycycline/*therapeutic use ; Elephantiasis, Filarial/diagnostic imaging/*drug therapy ; Humans ; Male ; Middle Aged ; Testicular Hydrocele/diagnostic imaging ; Treatment Outcome ; Ultrasonic Therapy ; Ultrasonography ; Wolbachia/*isolation & purification ; }, } @article {pmid18539347, year = {2008}, author = {Raverdy, S and Foster, JM and Roopenian, E and Carlow, CK}, title = {The Wolbachia endosymbiont of Brugia malayi has an active pyruvate phosphate dikinase.}, journal = {Molecular and biochemical parasitology}, volume = {160}, number = {2}, pages = {163-166}, doi = {10.1016/j.molbiopara.2008.04.014}, pmid = {18539347}, issn = {0166-6851}, mesh = {Amino Acid Sequence ; Animals ; Brugia malayi/*microbiology ; Cloning, Molecular ; Gene Expression ; Molecular Sequence Data ; Phylogeny ; Pyruvate, Orthophosphate Dikinase/genetics/*metabolism ; Sequence Alignment ; Wolbachia/*enzymology/genetics ; }, abstract = {Genome analysis of the glycolytic/gluconeogenic pathway in the Wolbachia endosymbiont from the filarial parasite Brugia malayi (wBm) has revealed that wBm lacks pyruvate kinase (PK) and may instead utilize the enzyme pyruvate phosphate dikinase (PPDK; ATP:pyruvate, orthophosphate phosphotransferase, EC 2.7.9.1). PPDK catalyses the reversible conversion of AMP, PPi and phosphoenolpyruvate (PEP) into ATP, Pi and pyruvate. The glycolytic pathway of most organisms, including mammals, contains exclusively PK for the production of pyruvate from PEP. Therefore, the absence of PPDK in mammals makes the enzyme an attractive Wolbachia drug target. In the present study, we have cloned and expressed an active wBm-PPDK, thereby providing insight into the energy metabolism of the endosymbiont. Our results support the development of wBm-PPDK as a promising new drug target in an anti-symbiotic approach to controlling filarial infection.}, } @article {pmid18538871, year = {2008}, author = {Keiser, PB and Coulibaly, Y and Kubofcik, J and Diallo, AA and Klion, AD and Traoré, SF and Nutman, TB}, title = {Molecular identification of Wolbachia from the filarial nematode Mansonella perstans.}, journal = {Molecular and biochemical parasitology}, volume = {160}, number = {2}, pages = {123-128}, pmid = {18538871}, issn = {0166-6851}, support = {Z01 AI000512-20/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/isolation & purification ; DNA Primers/genetics ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Genes, rRNA ; Mansonella/genetics/*microbiology ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 5S/genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Wolbachia/*classification/*genetics/isolation & purification ; }, abstract = {Wolbachiae are bacterial endosymbionts of insects and many filarial nematodes whose products trigger inflammation in filarial infections. The dependence of the parasites on their endosymbionts has also led to the use of antibiotics directed against the Wolbachiae, therapy that has been demonstrated to have a profound salutary effect on filarial infections. The identification of Wolbachiae in Mansonella species has been conclusively shown for Mansonella ozzardi (Mo), but not for Mansonella perstans (Mp). Using primers known to amplify the 16S ribosomal DNA of other filarial Wolbachiae, an identical 1393bp band was found in all samples tested. Sequence analysis of these samples demonstrated a single consensus sequence for Mp Wolbachia 16S rDNA that was most similar to Wolbachia sequences from other filarial nematodes. When aligned with the only other Mansonella Wolbachia sequence (Mo) there were only 8 nucleotide differences in the 1369bp overlapping sequence. Phylogenetic dendrograms, examining the relationship of the Mp Wolbachia to other Wolbachia 16S rDNA, showed that the Wolbachia tracked almost identically to the 5S rRNA of their parasite host. Wolbachia surface protein (WSP) was also demonstrated in protein extracted from Mp-containing whole blood. In advance of a treatment trial of Mp, a method for the quantitation of Mp Wolbachia was developed and used to demonstrate not only a relationship between microfilarial numbers and Wolbachia copy numbers, but also to demonstrate the effect of antibiotic on ridding Mp of Wolbachia.}, } @article {pmid18533441, year = {2008}, author = {Matsumoto, K and Izri, A and Dumon, H and Raoult, D and Parola, P}, title = {First detection of Wolbachia spp., including a new genotype, in sand flies collected in Marseille, France.}, journal = {Journal of medical entomology}, volume = {45}, number = {3}, pages = {466-469}, doi = {10.1603/0022-2585(2008)45[466:fdowsi]2.0.co;2}, pmid = {18533441}, issn = {0022-2585}, mesh = {Animals ; France ; Genes, Bacterial ; Genotype ; Phylogeny ; Polymerase Chain Reaction ; Psychodidae/*microbiology ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Wolbachia/classification/*genetics/*isolation & purification ; }, abstract = {Pooled sand fly samples collected in Marseille, France, in July 2005 were examined by polymerase chain reaction amplification and sequencing of the 16S rRNA gene of the Anaplasmataceae. Two different sequences of the 16S rRNA gene were obtained that are similar to bacteria belonging to the genus Wolbachia. These findings were confirmed by amplification, sequencing, and phylogenetic analysis of the wsp gene of the genus Wolbachia. The Wolbachia species from Sergentomyia minuta has been named "Wolbachia sp. detected in Se. minuta".}, } @article {pmid18510020, year = {2008}, author = {Cook, PE and McMeniman, CJ and O'Neill, SL}, title = {Modifying insect population age structure to control vector-borne disease.}, journal = {Advances in experimental medicine and biology}, volume = {627}, number = {}, pages = {126-140}, doi = {10.1007/978-0-387-78225-6_11}, pmid = {18510020}, issn = {0065-2598}, mesh = {Age Factors ; Animals ; Biological Evolution ; Disease Transmission, Infectious ; Fungi/isolation & purification/pathogenicity ; *Insect Vectors ; Insecta/*growth & development/microbiology ; Parvoviridae/isolation & purification/pathogenicity ; Temperature ; Wolbachia/isolation & purification/pathogenicity ; }, abstract = {Age is a critical determinant of the ability of most arthropod vectors to transmit a range of human pathogens. This is due to the fact that most pathogens require a period of extrinsic incubation in the arthropod host before pathogen transmission can occur. This developmental period for the pathogen often comprises a significant proportion of the expected lifespan of the vector. As such, only a small proportion of the population that is oldest contributes to pathogen transmission. Given this, strategies that target vector age would be expected to obtain the most significant reductions in the capacity of a vector population to transmit disease. The recent identification of biological agents that shorten vector lifespan, such as Wolbachia, entomopathogenic fungi and densoviruses, offer new tools for the control of vector-borne diseases. Evaluation of the efficacy of these strategies under field conditions will be possible due to recent advances in insect age-grading techniques. Implementation of all of these strategies will require extensive field evaluation and consideration of the selective pressures that reductions in vector longevity may induce on both vector and pathogen.}, } @article {pmid18510019, year = {2008}, author = {Rasgon, JL}, title = {Using predictive models to optimize Wolbachia-based strategies for vector-borne disease control.}, journal = {Advances in experimental medicine and biology}, volume = {627}, number = {}, pages = {114-125}, doi = {10.1007/978-0-387-78225-6_10}, pmid = {18510019}, issn = {0065-2598}, mesh = {Animals ; *Insect Vectors ; *Models, Theoretical ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {The development of resistance to insecticides by vector arthropods, the evolution of resistance to chemotherapeutic agents by parasites and the lack of clinical cures or vaccines for many diseases has stimulated a high-profile effort to develop vector-borne disease control strategies based on release of genetically-modified mosquitoes. Because transgenic insects are likely to be less fit than their wild-type counterparts, transgenic traits must be actively driven into the population in spite of fitness costs (population replacement). Wolbachia are maternally-inherited symbionts that are associated with numerous alterations in host reproductive biology. By a variety of mechanisms, Wolbachia-infected females have a reproductive advantage relative to uninfected females, allowing infection to spread rapidly through host populations to high frequency in spite of fitness costs. In theory, Wolbachia can be exploited to drive costly transgenes into vector populations for disease control. Before conducting an actual release, it is important to be able to predict how released Wolbachia infections are expected to behave. While inferences can be made by observing the dynamics of naturally-occurring infections, there is no ideal way to empirically test the efficacy ofa Wolbachia gene driver under field conditions prior to the first actual release. Mathematical models are a powerful way to predict the outcomes of transgenic insect releases and allow one to identify knowledge gaps, identify parameters that are critical to the success of releases, conduct risk-assessment analysis and investigate worst-case scenarios, and ultimately identify the most effective, most logistically feasible control method or methods. In this chapter, I review current and historical advances in applied models of Wolbachia spread, specifically within the context of applied population replacement strategies for vector-borne disease control.}, } @article {pmid18510018, year = {2008}, author = {Bourtzis, K}, title = {Wolbachia-based technologies for insect pest population control.}, journal = {Advances in experimental medicine and biology}, volume = {627}, number = {}, pages = {104-113}, doi = {10.1007/978-0-387-78225-6_9}, pmid = {18510018}, issn = {0065-2598}, mesh = {Animals ; Insecta/*growth & development ; Population Growth ; Virulence ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia are a group of obligatory intracellular and maternally inherited bacteria found in many arthropod species, including insects, mites, spiders, springtails, crustaceans, as well as in certain nematodes. Several PCR-based surveys suggest that over 20% of the arthropod species may be Wolbachia-infected, rendering this bacterium the most ubiquitous intracellular symbiont yet described. Wolbachia have recently attracted attention for their potential as novel and environmentally friendly bio-control agents. Wolbachia are able to invade and maintain themselves in the arthropod species through manipulation of the host's reproduction. Several strategies can be distinguished, one of which is cytoplasmic incompatibility (CI). Wolbachia-induced cytoplasmic incompatibility can be used beneficially in the following ways: (a) as a tool for insect pest population control in a way analogous to the "Sterile Insect technique" (SIT) and (b) as a drive system to spread desirable genotypes in field arthropod populations. In addition, virulent Wolbachia strains offer the potential to control vector species by modifying their population age structure. In the present chapter, I summarize the recent developments in Wolbachia research with an emphasis on the applied biology of Wolbachia and conclude with the challenges that Wolbachia researchers will face if they want to use and/or introduce Wolbachia into pest and vector species ofeconomic, environmental and public health relevance and, through Wolbachia-based technologies, to suppress or modify natural populations.}, } @article {pmid18502862, year = {2008}, author = {Rancès, E and Voronin, D and Tran-Van, V and Mavingui, P}, title = {Genetic and functional characterization of the type IV secretion system in Wolbachia.}, journal = {Journal of bacteriology}, volume = {190}, number = {14}, pages = {5020-5030}, pmid = {18502862}, issn = {1098-5530}, mesh = {Animals ; Bacterial Proteins/genetics/metabolism ; Carrier Proteins/*genetics/*metabolism ; Cloning, Molecular ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; Drosophila/microbiology ; Escherichia coli/genetics ; Female ; Gene Expression Profiling ; Gene Order ; Genes, Bacterial ; Hymenoptera/microbiology ; Microscopy, Fluorescence ; Molecular Sequence Data ; Open Reading Frames ; Operon ; Ovary/microbiology ; Protein Transport ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Virulence Factors/*genetics/*metabolism ; Wolbachia/*genetics/isolation & purification/*metabolism ; }, abstract = {A type IV secretion system (T4SS) is used by many symbiotic and pathogenic intracellular bacteria for the successful infection of and survival, proliferation, and persistence within hosts. In this study, the presence and function of the T4SS in Wolbachia strains were investigated by a combination of genetic screening and immunofluorescence microscopy. Two operons of virB-virD4 loci were found in the genome of Wolbachia pipientis strain wAtab3, from the Hymenoptera Asobara tabida, and strain wRi, infecting Drosophila simulans. One operon consisted of five vir genes (virB8, virB9, virB10, virB11, and virD4) and the downstream wspB locus. The other operon was composed of three genes (virB3, virB4, and virB6) and included four additional open reading frames (orf1 to orf4) orientated in the same direction. In cell culture and insect hosts infected with different Wolbachia strains, the bona fide vir genes were polycistronically transcribed, together with the downstream adjacent loci, notably, as virB8 to virD4 and wspB and as virB3, virB4, virB6, and orf1 to orf4. Two peptides encompassing conserved C and N termini of the Wolbachia VirB6 protein were used for the production of polyclonal antibodies. Anti-VirB6 antibodies could detect the corresponding recombinant protein by chemifluorescence on Western blots of total proteins from Escherichia coli transformants and Wolbachia strains cultured in cell lines. Using immunofluorescence microscopy, we further demonstrated that the VirB6 protein was produced by Wolbachia strains in ovaries of insects harboring wAtab3 or wRi and cell lines infected with wAlbB or wMelPop. As VirB6 is known to associate with other VirB proteins to form a membrane-spanning structure, this finding suggests that a T4SS may function in Wolbachia.}, } @article {pmid18495345, year = {2008}, author = {Hornok, S and Földvári, G and Elek, V and Naranjo, V and Farkas, R and de la Fuente, J}, title = {Molecular identification of Anaplasma marginale and rickettsial endosymbionts in blood-sucking flies (Diptera: Tabanidae, Muscidae) and hard ticks (Acari: Ixodidae).}, journal = {Veterinary parasitology}, volume = {154}, number = {3-4}, pages = {354-359}, doi = {10.1016/j.vetpar.2008.03.019}, pmid = {18495345}, issn = {0304-4017}, mesh = {Anaplasma marginale/*genetics ; Anaplasmosis/microbiology/transmission ; Animals ; Cattle ; Cattle Diseases/transmission ; Insect Vectors/microbiology ; Ixodidae/*microbiology ; Muscidae/*microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*isolation & purification ; }, abstract = {In an attempt to identify the main vector and possible transmission routes of Anaplasma spp. in a region of Hungary with high prevalence of ovine and bovine anaplasmosis, DNA was extracted from 316 haematophagous arthropods (individually or in pools), including 4 species of ixodid ticks, 6 species of tabanid flies and hornflies. Midichloria-like organisms were identified with PCR (amplifying a portion of the 16S rRNA gene) and sequencing from Dermacentor marginatus and Ixodes ricinus. Significantly higher 16S positive D. marginatus individuals were collected in March than in April, suggesting earlier questing of ticks that contain rickettsial agents (thus endosymbionts). Midichloria- and Wolbachia-like organisms were also found in randomly caught horse flies (Tabanus bovinus and T. tergestinus) as well as hornflies (Haematobia irritans), respectively, with 97-99% similarity to sequences deposited in the GenBank. Although all ticks were negative in the Anaplasma spp.-specific msp4 PCR, four individuals of T. bovinus collected near to grazing cattle were positive for Anaplasma marginale. The results of the present study provide the first molecular evidence for the potential mechanical vector role of T. bovinus in the transmission of A. marginale, and broaden the range of haematophagous arthropods harbouring Midichloria-like bacteria, for the first time in any Dermacentor or Tabanus species.}, } @article {pmid18494796, year = {2008}, author = {Pham, VD and Konstantinidis, KT and Palden, T and DeLong, EF}, title = {Phylogenetic analyses of ribosomal DNA-containing bacterioplankton genome fragments from a 4000 m vertical profile in the North Pacific Subtropical Gyre.}, journal = {Environmental microbiology}, volume = {10}, number = {9}, pages = {2313-2330}, doi = {10.1111/j.1462-2920.2008.01657.x}, pmid = {18494796}, issn = {1462-2920}, mesh = {Bacteria/*genetics ; Computational Biology ; DNA, Ribosomal/genetics ; Gene Library ; Genes, rRNA ; *Genome, Bacterial ; Nucleic Acid Hybridization/methods ; Pacific Ocean ; Phylogeny ; RNA, Bacterial/*genetics ; Seawater/microbiology ; Sequence Alignment ; Sequence Analysis, DNA ; Software ; *Water Microbiology ; }, abstract = {High-throughput identification of rRNA gene-containing clones in large insert metagenomic libraries is difficult, because of the high background of host ribosomal RNA (rRNA) and rRNA genes. To address this challenge, a membrane hybridization method was developed to identify all bacterial small subunit rRNA-containing fosmid clones of microbial community DNA from seven different depths in the North Pacific Subtropical Gyre. Out of 101,376 clones screened, 751 rDNA-containing clones were identified that grouped in approximately 60 different clades. Several rare sequences only remotely related to known groups were detected, including a Wolbachia-related sequence containing a putative intron or intervening sequence, as well as seven sequences from Order Myxococcales not previously detected in pelagic habitats. Stratified, depth-specific population structure was evident within both cultured and uncultured lineages. Conversely, some eurybathyal members of the genera Alcanivorax and Rhizobium shared identical small subunit ribosomal DNA sequences that were distributed from surface waters to the 4000 m depth. Comparison with similar analyses in Monterey Bay microbial communities revealed previously recognized, as well as some distinctive, depth-stratified partitioning that distinguished coastal from open ocean bacterioplankton populations. While some bias was evident in fosmid clone recovery in a few particular lineages, the overall phylogenetic group recovery and distributions were consistent with previous studies, as well as with direct shotgun sequence data from the same source DNA.}, } @article {pmid18489749, year = {2008}, author = {Ros, VI and Breeuwer, JA and Menken, SB}, title = {Origins of asexuality in Bryobia mites (Acari: Tetranychidae).}, journal = {BMC evolutionary biology}, volume = {8}, number = {}, pages = {153}, pmid = {18489749}, issn = {1471-2148}, mesh = {Animals ; Base Composition ; DNA Primers ; DNA, Ribosomal/genetics ; Evolution, Molecular ; *Genetic Variation ; Likelihood Functions ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 28S/genetics ; Reproduction, Asexual/*genetics ; Sequence Analysis, DNA ; Species Specificity ; Tetranychidae/classification/genetics/*physiology ; }, abstract = {BACKGROUND: Obligate asexual reproduction is rare in the animal kingdom. Generally, asexuals are considered evolutionary dead ends that are unable to radiate. The phytophagous mite genus Bryobia contains a large number of asexual species. In this study, we investigate the origin and evolution of asexuality using samples from 111 populations in Europe, South Africa and the United States, belonging to eleven Bryobia species. We also examine intraspecific clonal diversity for one species, B. kissophila, by genotyping individuals from 61 different populations. Knowledge on the origin of asexuality and on clonal diversity can contribute to our understanding of the paradox of sex.

RESULTS: The majority (94%) of 111 sampled populations reproduces asexually. Analysis of part of nuclear 28S rDNA shows that these asexuals do not form a monophyletic clade. Analysis of the mitochondrial COI region shows that intraspecific variation is extensive (up to 8.8%). Within B. kissophila, distinct clades are found, which are absent at the nuclear 28S rDNA level. Moreover, paraphyletic patterns are found at the mitochondrial DNA.

CONCLUSION: Asexuality is widespread in the genus Bryobia, signifying that some animal taxa do contain a high number of asexuals. We argue that asexuality originated multiple times within Bryobia. Wolbachia bacteria cause asexuality in at least two Bryobia species and may have infected different species independently. The high intraspecific clonal diversity and the patterns of paraphyly at the mitochondrial DNA in B. kissophila might be explained by a high mutation fixation rate and past hybridization events. Reproductive parasites like Wolbachia and Cardinium might influence these processes. We discuss the role these bacteria could play in the evolutionary success of asexual species.}, } @article {pmid18486691, year = {2008}, author = {McCall, JW and Genchi, C and Kramer, LH and Guerrero, J and Venco, L}, title = {Heartworm disease in animals and humans.}, journal = {Advances in parasitology}, volume = {66}, number = {}, pages = {193-285}, doi = {10.1016/S0065-308X(08)00204-2}, pmid = {18486691}, issn = {0065-308X}, mesh = {Animals ; Cat Diseases/diagnosis/epidemiology/parasitology/prevention & control ; Cats ; Dirofilaria immitis/immunology/*pathogenicity ; *Dirofilariasis/diagnosis/epidemiology/parasitology/prevention & control ; Dog Diseases/diagnosis/epidemiology/parasitology/prevention & control ; Dogs ; Female ; Ferrets ; Filaricides/therapeutic use ; *Host-Parasite Interactions ; Humans ; Male ; Symbiosis ; Wolbachia/growth & development ; }, abstract = {Heartworm disease due to Dirofilaria immitis continues to cause severe disease and even death in dogs and other animals in many parts of the world, even though safe, highly effective and convenient preventatives have been available for the past two decades. Moreover, the parasite and vector mosquitoes continue to spread into areas where they have not been reported previously. Heartworm societies have been established in the USA and Japan and the First European Dirofilaria Days (FEDD) Conference was held in Zagreb, Croatia, in February of 2007. These organizations promote awareness, encourage research and provide updated guidelines for the diagnosis, treatment and prevention of heartworm disease. The chapter begins with a review of the biology and life cycle of the parasite. It continues with the prevalence and distribution of the disease in domestic and wild animals, with emphasis on more recent data on the spreading of the disease and the use of molecular biology techniques in vector studies. The section on pathogenesis and immunology also includes a discussion of the current knowledge of the potential role of the Wolbachia endosymbiont in inflammatory and immune responses to D. immitis infection, diagnostic use of specific immune responses to the bacteria, immunomodulatory activity and antibiotic treatment of infected animals. Canine, feline and ferret heartworm disease are updated with regard to the clinical presentation, diagnosis, prevention, therapy and management of the disease, with special emphasis on the recently described Heartworm Associated Respiratory Disease (HARD) Syndrome in cats. The section devoted to heartworm infection in humans also includes notes on other epizootic filariae, particularly D. repens in humans in Europe. The chapter concludes with a discussion on emerging strategies in heartworm treatment and control, highlighting the potential role of tetracycline antibiotics in adulticidal therapy.}, } @article {pmid18478054, year = {2008}, author = {Suh, JM and Stenesen, D and Peters, JM and Inoue, A and Cade, A and Graff, JM}, title = {An RGS-containing sorting nexin controls Drosophila lifespan.}, journal = {PloS one}, volume = {3}, number = {5}, pages = {e2152}, pmid = {18478054}, issn = {1932-6203}, mesh = {5' Untranslated Regions ; Animals ; Animals, Genetically Modified ; Carrier Proteins/chemistry/*physiology ; Drosophila/genetics/*physiology ; Fat Body/physiology ; Larva/physiology ; *Longevity ; Mutation ; Oligopeptides/*chemistry ; Sorting Nexins ; Vesicular Transport Proteins/chemistry/*physiology ; }, abstract = {The pursuit of eternal youth has existed for centuries and recent data indicate that fat-storing tissues control lifespan. In a D. melanogaster fat body insertional mutagenic enhancer trap screen designed to isolate genes that control longevity, we identified a regulator of G protein signaling (RGS) domain containing sorting nexin, termed snazarus (sorting nexin lazarus, snz). Flies with insertions into the 5' UTR of snz live up to twice as long as controls. Transgenic expression of UAS-Snz from the snz Gal4 enhancer trap insertion, active in fat metabolic tissues, rescued lifespan extension. Further, the lifespan extension of snz mutants was independent of endosymbiont, e.g., Wolbachia, effects. Notably, old snz mutant flies remain active and fertile indicating that snz mutants have prolonged youthfulness, a goal of aging research. Since mammals have snz-related genes, it is possible that the functions of the snz family may be conserved to humans.}, } @article {pmid18461124, year = {2008}, author = {Brennan, LJ and Keddie, BA and Braig, HR and Harris, HL}, title = {The endosymbiont Wolbachia pipientis induces the expression of host antioxidant proteins in an Aedes albopictus cell line.}, journal = {PloS one}, volume = {3}, number = {5}, pages = {e2083}, pmid = {18461124}, issn = {1932-6203}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Bacterial Proteins/genetics/isolation & purification ; Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Flow Cytometry ; *Gene Expression Regulation ; Glutathione Peroxidase/genetics ; Insect Proteins/*genetics/isolation & purification ; Polymerase Chain Reaction ; Reactive Oxygen Species/metabolism ; Rifampin/pharmacology ; Superoxide Dismutase/genetics ; Symbiosis ; Wolbachia/drug effects/genetics/*physiology ; }, abstract = {Wolbachia are obligate intracellular bacteria which commonly infect arthropods. They are maternally inherited and capable of altering host development, sex determination, and reproduction. Reproductive manipulations include feminization, male-killing, parthenogenesis, and cytoplasmic incompatibility. The mechanism by which Wolbachia avoid destruction by the host immune response is unknown. Generation of antimicrobial peptides (AMPs) and reactive oxygen species (ROS) by the host are among the first lines of traditional antimicrobial defense. Previous work shows no link between a Wolbachia infection and the induction of AMPs. Here we compare the expression of protein in a cell line naturally infected with Wolbachia and an identical cell line cured of the infection through the use of antibiotics. Protein extracts of each cell line were analyzed by two dimensional gel electrophoresis and LC/MS/MS. Our results show the upregulation of host antioxidant proteins, which are active against ROS generated by aerobic cell metabolism and during an immune response. Furthermore, flow cytometric and microscopic analysis demonstrates that ROS production is significantly greater in Wolbachia-infected mosquito cells and is associated with endosymbiont-containing vacuoles located in the host cell cytoplasm. This is the first empirical data supporting an association between Wolbachia and the insect antioxidant system.}, } @article {pmid18458997, year = {2008}, author = {Kageyama, D and Narita, S and Noda, H}, title = {Transfection of feminizing Wolbachia endosymbionts of the butterfly, Eurema hecabe, into the cell culture and various immature stages of the silkmoth, Bombyx mori.}, journal = {Microbial ecology}, volume = {56}, number = {4}, pages = {733-741}, pmid = {18458997}, issn = {0095-3628}, mesh = {Animals ; Bombyx/cytology/genetics/*microbiology ; Butterflies/*microbiology ; Cell Line ; Female ; Male ; Oocytes/metabolism ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Wolbachia/genetics/*growth & development ; }, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria of invertebrates that can manipulate the reproductive systems of their arthropod hosts in a variety of ways. To establish a useful model system for investigating the mechanism of Wolbachia-induced host feminization, we conducted the following series of experiments: (1) feminizing Wolbachia of the butterfly, Eurema hecabe, were transferred into cell cultures of the silkmoth, Bombyx mori, and (2) the transfected Wolbachia in cell cultures were inoculated into B. mori at four immature stages. Wolbachia were successfully transfected into the cell cultures and stably maintained for more than 1 year (>30 passages). However, none of the inoculated insects produced mature oocytes that were Wolbachia-positive. This finding was consistent with the fact that Wolbachia was not detected in individuals in subsequent generations. In contrast, Wolbachia were detected at relatively high frequencies (60-80% of individuals) in the somatic tissues of inoculated insects. Real-time quantitative polymerase chain reaction revealed that the Wolbachia densities in the cultured cells were approximately tenfold higher than those in the native host E. hecabe. Among B. mori individuals inoculated at various developmental stages, those inoculated at early stages exhibited higher Wolbachia densities at the adult stage. The Wolbachia densities in individuals inoculated at the second-instar stage were comparable to those in intact E. hecabe. These results suggest that infection and/or proliferation of Wolbachia in germline cells are actively hindered by regulation in B. mori but feasible in somatic cells and that the Wolbachia densities in somatic tissues are regulated by the living host insects.}, } @article {pmid18456851, year = {2008}, author = {Peng, Y and Nielsen, JE and Cunningham, JP and McGraw, EA}, title = {Wolbachia infection alters olfactory-cued locomotion in Drosophila spp.}, journal = {Applied and environmental microbiology}, volume = {74}, number = {13}, pages = {3943-3948}, pmid = {18456851}, issn = {1098-5336}, mesh = {Animals ; Drosophila/classification/*microbiology/*physiology ; Drosophila melanogaster/microbiology/physiology ; *Host-Pathogen Interactions ; Locomotion/*physiology ; Smell/*physiology ; Species Specificity ; Symbiosis ; Wolbachia/classification/*pathogenicity ; }, abstract = {Wolbachia pipientis is an endosymbiotic bacterium present in diverse insect species. Although it is well studied for its dramatic effects on host reproductive biology, little is known about its effects on other aspects of host biology, despite its presence in a wide array of host tissues. This study examined the effects of three Wolbachia strains on two different Drosophila species, using a laboratory performance assay for insect locomotion in response to olfactory cues. The results demonstrate that Wolbachia infection can have significant effects on host responsiveness that vary with respect to the Wolbachia strain-host species combination. The wRi strain, native to Drosophila simulans, increases the basal activity level of the host insect as well as its responsiveness to food cues. In contrast, the wMel strain and the virulent wMelPop strain, native to Drosophila melanogaster, cause slight decreases in responsiveness to food cues but do not alter basal activity levels in the host. Surprisingly, the virulent wMelPop strain has very little impact on host responsiveness in D. simulans. This novel strain-host relationship was artificially created previously by transinfection. These findings have implications for understanding the evolution and spread of Wolbachia infections in wild populations and for Wolbachia-based vector-borne disease control strategies currently being developed.}, } @article {pmid18439691, year = {2008}, author = {Hewitson, JP and Harcus, YM and Curwen, RS and Dowle, AA and Atmadja, AK and Ashton, PD and Wilson, A and Maizels, RM}, title = {The secretome of the filarial parasite, Brugia malayi: proteomic profile of adult excretory-secretory products.}, journal = {Molecular and biochemical parasitology}, volume = {160}, number = {1}, pages = {8-21}, doi = {10.1016/j.molbiopara.2008.02.007}, pmid = {18439691}, issn = {0166-6851}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Amino Acid Sequence ; Animals ; Blotting, Western ; Brugia malayi/*chemistry ; Chromatography, Liquid ; Computational Biology ; Electrophoresis, Gel, Two-Dimensional ; Filariasis/parasitology ; Galectins/analysis ; Helminth Proteins/*analysis/*metabolism ; Humans ; Macrophage Migration-Inhibitory Factors/analysis ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases/analysis ; Proteome/*analysis/*metabolism ; Proteomics/methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; }, abstract = {The secretome of a parasite in its definitive host can be considered to be its genome in trans, to the extent that secreted products encoded by the parasite fulfill their function in the host milieu. The 'extended phenotype' of the filarial parasite, Brugia malayi, is of particular interest because of the evidence that infection results in potent down-modulation of the host immune response. We collected B. malayi 'excretory-secretory' (BES) proteins from adult parasites and using a combination of shotgun LC-MS/MS and 2D gel electrophoresis, identified 80 B. malayi and two host proteins in BES, of which 31 (38%) were detectable in whole worm extract (BmA). Products which were enriched in BES relative to BmA included phosphatidylethanolamine-binding protein (PEB), leucyl aminopeptidase (LAP, homologue of ES-62 from the related filaria Acanthocheilonema viteae), N-acetylglucosaminyltransferase (GlcNAcT) and galectin-1, in addition to the previously described major surface glycoprotein, glutathione peroxidase (gp29, GPX-1) and the cytokine homologue macrophage migration inhibitory factor (MIF-1). One of the most abundant released proteins was triose phosphate isomerase (TPI), yet many other glycolytic enzymes (such as aldolase and GAPDH) were found only in the somatic extract. Among the more prominent novel products identified in BES were a set of 11 small transthyretin-like proteins, and three glutamine-rich-repeat mucin-like proteins. Notably, no evidence was found of any secreted protein corresponding to the genome of the Wolbachia endosymbiont present in B. malayi. Western blotting with anti-phosphorylcholine (PC) monoclonal antibody identified that GlcNAcT, and not the ES-62 homologue, is the major PC-bearing protein in BES, while probing with human filariasis sera showed preferential reactivity to galectin-1 and to processed forms of myotactin. Overall, this analysis demonstrates selective release of a suite of newly identified proteins not previously suspected to be involved at the host-parasite interface, and provides important new perspectives on the biology of the filarial parasite.}, } @article {pmid18433753, year = {2008}, author = {Bazzocchi, C and Mortarino, M and Grandi, G and Kramer, LH and Genchi, C and Bandi, C and Genchi, M and Sacchi, L and McCall, JW}, title = {Combined ivermectin and doxycycline treatment has microfilaricidal and adulticidal activity against Dirofilaria immitis in experimentally infected dogs.}, journal = {International journal for parasitology}, volume = {38}, number = {12}, pages = {1401-1410}, doi = {10.1016/j.ijpara.2008.03.002}, pmid = {18433753}, issn = {1879-0135}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Antigens, Helminth/immunology ; Dirofilaria immitis/*drug effects/immunology ; Dirofilariasis/*drug therapy/immunology ; Dog Diseases/drug therapy ; Dogs ; Doxycycline/immunology/*therapeutic use ; Filaricides/immunology/*therapeutic use ; Immunohistochemistry ; Ivermectin/*therapeutic use ; Microfilariae/isolation & purification ; Polymerase Chain Reaction ; Wolbachia/drug effects/isolation & purification ; }, abstract = {There is still a pressing need for effective adulticide treatment for human and animal filarial infections. Like many filarial nematodes, Dirofilaria immitis, the causative agent of canine heartworm disease, harbours the bacterial endosymbiont Wolbachia, which has been shown to be essential for worm development, fecundity and survival. Here the authors report the effect of different treatment regimens in dogs experimentally infected with adult D. immitis on microfilariemia, antigenemia, worm recovery and Wolbachia content. Treatment with ivermectin (IVM; 6 microg/kg per os weekly) combined with doxycycline (DOXY; 10 mg/kg/day orally from Weeks 0-6, 10-12, 16-18, 22-26 and 28-34) resulted in a significantly faster decrease of circulating microfilariae and higher adulticidal activity compared with either IVM or DOXY alone. Quantitative PCR analysis of ftsZ (Wolbachia DNA) and 18S rDNA (nematode DNA) absolute copy numbers showed significant decreases in Wolbachia content compared with controls in worms recovered from DOXY-treated dogs that were not, however, associated with worm death. Worms from IVM/DOXY-treated dogs, on the other hand, had Wolbachia/nematode DNA ratios similar to those of control worms, suggesting a loss of both Wolbachia and nematode DNA as indicated by absolute copy number values. Histology and transmission electron microscopy of worms recovered from the IVM/DOXY combination group showed complete loss of uterine content in females and immunohistochemistry for Wolbachia was negative. Results indicate that the combination of these two drugs causes adult worm death. This could have important implications for control of human and animal filarial infections.}, } @article {pmid18430940, year = {2008}, author = {Zabalou, S and Apostolaki, A and Pattas, S and Veneti, Z and Paraskevopoulos, C and Livadaras, I and Markakis, G and Brissac, T and Merçot, H and Bourtzis, K}, title = {Multiple rescue factors within a Wolbachia strain.}, journal = {Genetics}, volume = {178}, number = {4}, pages = {2145-2160}, pmid = {18430940}, issn = {0016-6731}, mesh = {Animals ; Bacterial Typing Techniques ; Cytoplasm/microbiology ; Drosophila/classification/microbiology ; Embryo, Nonmammalian/cytology/microbiology ; Female ; Fluorescent Antibody Technique ; *Genes, Bacterial ; Linear Models ; Male ; Phylogeny ; Rickettsiaceae Infections/microbiology ; Wolbachia/classification/cytology/*genetics ; }, abstract = {Wolbachia-induced cytoplasmic incompatibility (CI) is expressed when infected males are crossed with either uninfected females or females infected with Wolbachia of different CI specificity. In diploid insects, CI results in embryonic mortality, apparently due to the the loss of the paternal set of chromosomes, usually during the first mitotic division. The molecular basis of CI has not been determined yet; however, several lines of evidence suggest that Wolbachia exhibits two distinct sex-dependent functions: in males, Wolbachia somehow "imprints" the paternal chromosomes during spermatogenesis (mod function), whereas in females, the presence of the same Wolbachia strain(s) is able to restore embryonic viability (resc function). On the basis of the ability of Wolbachia to induce the modification and/or rescue functions in a given host, each bacterial strain can be classified as belonging in one of the four following categories: mod(+) resc(+), mod(-) resc(+), mod(-) resc(-), and mod(+) resc(-). A so-called "suicide" mod(+) resc(-) strain has not been found in nature yet. Here, a combination of embryonic cytoplasmic injections and introgression experiments was used to transfer nine evolutionary, distantly related Wolbachia strains (wYak, wTei, wSan, wRi, wMel, wHa, wAu, wNo, and wMa) into the same host background, that of Drosophila simulans (STCP strain), a highly permissive host for CI expression. We initially characterized the modification and rescue properties of the Wolbachia strains wYak, wTei, and wSan, naturally present in the yakuba complex, upon their transfer into D. simulans. Confocal microscopy and multilocus sequencing typing (MLST) analysis were also employed for the evaluation of the CI properties. We also tested the compatibility relationships of wYak, wTei, and wSan with all other Wolbachia infections. So far, the cytoplasmic incompatibility properties of different Wolbachia variants are explained assuming a single pair of modification and rescue factors specific to each variant. This study shows that a given Wolbachia variant can possess multiple rescue determinants corresponding to different CI systems. In addition, our results: (a) suggest that wTei appears to behave in D. simulans as a suicide mod(+) resc(-) strain, (b) unravel unique CI properties, and (c) provide a framework to understand the diversity and the evolution of new CI-compatibility types.}, } @article {pmid18421478, year = {2008}, author = {Hoerauf, A and Marfo-Debrekyei, Y and Büttner, M and Debrah, AY and Konadu, P and Mand, S and Adjei, O and Büttner, DW}, title = {Effects of 6-week azithromycin treatment on the Wolbachia endobacteria of Onchocerca volvulus.}, journal = {Parasitology research}, volume = {103}, number = {2}, pages = {279-286}, pmid = {18421478}, issn = {0932-0113}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/administration & dosage/*therapeutic use ; Azithromycin/administration & dosage/*therapeutic use ; Endemic Diseases ; Female ; Ghana/epidemiology ; Humans ; Male ; Middle Aged ; Onchocerca volvulus/*drug effects/growth & development/*microbiology ; Onchocerciasis/*drug therapy/epidemiology/parasitology ; Symbiosis ; Treatment Outcome ; Wolbachia/*drug effects ; }, abstract = {The effects of azithromycin treatment on the presence of Wolbachia endobacteria and on the embryogenesis and microfilariae production of Onchocerca volvulus were studied. In 2002, in an endemic area in Ghana, 37 onchocerciasis patients were treated for 6 weeks with azithromycin: 23 patients received 250 mg every day, and 14 took 1,200 mg once a week. After 6 and/or 12 months, all palpable worm nodules were extirpated from 31 treated and nine additional untreated patients, and the presence of Wolbachia and embryogenesis were assessed by immunohistology. In nodules taken 6 months after treatment with either dose and 12 months after 1,200 mg/week, the Wolbachia loads of the worms were not different from those of untreated worms. However, 12 months after the 250-mg/day azithromycin regimen, significantly less female worms (65% compared to 92% untreated ones) presented many Wolbachia, although the reduction was less pronounced than observed in other studies after treatment with doxycycline. Embryogenesis and microfilariae production were not reduced. It is concluded that azithromycin administered alone for 6 weeks at 250 mg/day or 1,200 mg/week is not suitable for treatment of human onchocerciasis. But daily azithromycin should be studied in combination with other drugs and with other doses.}, } @article {pmid18419441, year = {2008}, author = {Supali, T and Djuardi, Y and Pfarr, KM and Wibowo, H and Taylor, MJ and Hoerauf, A and Houwing-Duistermaat, JJ and Yazdanbakhsh, M and Sartono, E}, title = {Doxycycline treatment of Brugia malayi-infected persons reduces microfilaremia and adverse reactions after diethylcarbamazine and albendazole treatment.}, journal = {Clinical infectious diseases : an official publication of the Infectious Diseases Society of America}, volume = {46}, number = {9}, pages = {1385-1393}, doi = {10.1086/586753}, pmid = {18419441}, issn = {1537-6591}, mesh = {Adolescent ; Adult ; Aged ; Albendazole/adverse effects/*therapeutic use ; Animals ; Brugia malayi/*drug effects ; Child ; Diethylcarbamazine/adverse effects/*therapeutic use ; Double-Blind Method ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; Elephantiasis, Filarial/*drug therapy ; Female ; Filaricides/adverse effects/therapeutic use ; Humans ; Male ; Middle Aged ; Treatment Outcome ; }, abstract = {BACKGROUND: The efficacy of doxycycline for treating the causal agent of human lymphatic filariasis, Brugia malayi, is unknown. Standard treatment with diethylcarbamazine-albendazole is associated with adverse reactions. We assessed whether doxycycline alone or in combination with diethylcarbamazine-albendazole would lead to sustained amicrofilaremia and reduced incidence of adverse reactions.

METHODS: A double-blind, randomized, placebo-controlled 6-week field trial of doxycycline treatment (100 mg/day) of 161 persons infected with B. malayi was conducted. Four months after receiving doxycycline (n=119) or placebo (n=42), participants received diethylcarbamazine (6 mg/kg) plus albendazole (400 mg) or a matching placebo. Adverse reactions were assessed 48 and 60 h after administration of diethylcarbamazine-albendazole. Treatment efficacy was evaluated at 2, 4, and 12 months after the initial doxycycline treatment.

RESULTS: Four months after beginning doxycycline treatment, Wolbachia loads were reduced by 98%. Doxycycline treatment reduced the prevalence of microfilaremia at 2, 4, and 12 months of follow-up (P<.001 for all time points). At the 1-year follow-up, prevalence was reduced by 77% and 87.5% in patients receiving doxycycline alone or doxycycline plus diethylcarbamazine-albendazole, respectively. In contrast, the reduction of microfilaremia in the group receiving placebo doxycycline plus diethylcarbamazine-albendazole was merely 26.7%. Adverse reactions were lowest in the group receiving doxycycline plus placebo diethylcarbamazine-albendazole and highest in the group receiving placebo doxycycline plus diethylcarbamazine-albendazole. The proportion of persons with high fever and severe adverse reactions was significantly reduced in the group treated with doxycycline plus diethylcarbamazine-albendazole.

CONCLUSIONS: A 6-week course of doxycycline, either alone or in combination with diethylcarbamazine-albendazole, leads to a decrease in microfilaremia and reduces adverse reactions to antifilarial treatment in B. malayi-infected persons.}, } @article {pmid18405836, year = {2008}, author = {Hanington, PC and Brennan, LJ and Belosevic, M and Andrew Keddie, B}, title = {Molecular and functional characterization of granulin-like molecules of insects.}, journal = {Insect biochemistry and molecular biology}, volume = {38}, number = {5}, pages = {596-603}, doi = {10.1016/j.ibmb.2008.02.002}, pmid = {18405836}, issn = {0965-1748}, mesh = {Aedes/*metabolism ; Amino Acid Sequence ; Animals ; Cell Line ; Cell Proliferation ; Conserved Sequence ; Goldfish/*metabolism ; Hemocytes/metabolism ; Insect Proteins/isolation & purification/*metabolism ; Intercellular Signaling Peptides and Proteins/isolation & purification/*metabolism ; Manduca/*metabolism ; Molecular Sequence Data ; Phylogeny ; Progranulins ; Recombinant Proteins/metabolism ; Structural Homology, Protein ; }, abstract = {Granulins are a group of highly conserved growth factors that have been described from a variety of organisms spanning the metazoa. Here, we report on the identification of two partial transcripts encoding granulin-like molecules from Aa23 embryonic cells of Aedes albopictus and primary haemocytes from Manduca sexta. Both these partial transcripts had the characteristic 12-cysteine motif that is a hallmark of the granulin family and they represent the first granulin mRNA transcripts identified from insects. Moreover, we demonstrate that the recombinant granulin molecule that we originally cloned in the goldfish, induced proliferation of both Aa23 embryonic cells and primary haemocytes. Interestingly, this proliferative effect was upregulated in the presence of the intracellular symbiotic bacterium Wolbachia pipientis. Thus, granulin appears to be a highly conserved growth factor not only in lower vertebrates but also invertebrates.}, } @article {pmid18401667, year = {2008}, author = {Fallon, AM}, title = {Cytological properties of an Aedes albopictus mosquito cell line infected with Wolbachia strain wAlbB.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {44}, number = {5-6}, pages = {154-161}, pmid = {18401667}, issn = {1071-2690}, support = {R21 AI070913/AI/NIAID NIH HHS/United States ; R21 AI070913-02/AI/NIAID NIH HHS/United States ; AI070913/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*cytology/drug effects/growth & development/*microbiology ; Animals ; Cell Line ; Tetracycline/pharmacology ; Time Factors ; Wolbachia/drug effects/*physiology ; }, abstract = {In vitro production of the obligate intracellular bacterium, Wolbachia pipientis, is essential to its manipulation as a genetic tool to spread transgenes within vector populations. We have adapted the Wolbachia-infected Aa23 Aedes albopictus mosquito cell line to Eagle's minimal medium, supplemented with nonessential amino acids, glutamine, and 20% fetal bovine serum. When plated at low densities, Aa23E cells grew as patchy monolayers, comprised of non-contiguous clusters of cells that gave rise to solid clumps of tightly adherent cells. Multicellular clumps eventually detached from the substrate and floated freely in the medium. Removal of Wolbachia by treatment with tetracycline did not alter the cytological properties of the host cells, which had a population doubling time of 4-5 d. The presence of Wolbachia was monitored by Giemsa staining of cytological preparations, polymerase chain reaction (PCR) amplification of Wolbachia 16S ribosomal DNA, and by simultaneous PCR amplification of ribosomal protein genes from Wolbachia and mosquito host cell genomes. Wolbachia morphology was pleomorphic, and Wolbachia DNA persisted in the culture medium for several weeks after degradation of PCR-amplifiable host cell DNA.}, } @article {pmid18391545, year = {2007}, author = {Traut, W and Sahara, K and Marec, F}, title = {Sex chromosomes and sex determination in Lepidoptera.}, journal = {Sexual development : genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation}, volume = {1}, number = {6}, pages = {332-346}, doi = {10.1159/000111765}, pmid = {18391545}, issn = {1661-5433}, mesh = {Animals ; Bombyx/genetics ; Butterflies/genetics ; Cell Nucleus/genetics ; Crossing Over, Genetic/genetics ; Dosage Compensation, Genetic ; Evolution, Molecular ; Female ; Interphase ; Karyotyping ; Lepidoptera/*genetics ; Male ; Meiosis ; Moths/genetics ; Phylogeny ; Sex Chromatin/genetics ; Sex Chromosomes/*genetics ; *Sex Determination Processes ; }, abstract = {The speciose insect order Lepidoptera (moths and butterflies) and their closest relatives, Trichoptera (caddis flies), share a female-heterogametic sex chromosome system. Originally a Z/ZZ (female/male) system, it evolved by chromosome rearrangement to a WZ/ZZ (female/male) system in the most species-rich branch of Lepidoptera, a monophyletic group consisting of Ditrysia and Tischeriina, which together comprise more than 98% of all species. Further sporadic rearrangements created multi-sex chromosome systems; sporadic losses of the W changed the system formally back to Z/ZZ in some species. Primary sex determination depends on a Z-counting mechanism in Z/ZZ species, but on a female-determining gene, Fem, in the W chromosome of the silkworm. The molecular mechanism is unknown in both cases. The silkworm shares the last step, dsx, of the hierarchical sex-determining pathway with Drosophila and other insects investigated, but probably not the intermediate steps between the primary signal and dsx. The W chromosome is heterochromatic in most species. It contains few genes and is flooded with interspersed repetitive elements. In interphase nuclei of females it is readily discernible as a heterochromatic body which grows with increasing degree of polyploidy in somatic cells. It is used as a marker for the genetic sex in studies of intersexes and Wolbachia infections. The sex chromosome system is being exploited in economically important species. Special strains have been devised for mass rearing of male-only broods in the silkworm for higher silk production and in pest species for the release of sterile males in pest management programs.}, } @article {pmid18387819, year = {2008}, author = {Kurz, M and Iturbe-Ormaetxe, I and Jarrott, R and Cowieson, N and Robin, G and Jones, A and King, GJ and Frei, P and Glockshuber, R and O'Neill, SL and Heras, B and Martin, JL}, title = {Cloning, expression, purification and characterization of a DsbA-like protein from Wolbachia pipientis.}, journal = {Protein expression and purification}, volume = {59}, number = {2}, pages = {266-273}, doi = {10.1016/j.pep.2008.02.008}, pmid = {18387819}, issn = {1096-0279}, mesh = {Amino Acid Sequence ; Bacterial Proteins/*biosynthesis/*chemistry/isolation & purification ; Cloning, Molecular ; Escherichia coli/genetics ; Molecular Sequence Data ; Protein Disulfide-Isomerases/*biosynthesis/*chemistry/isolation & purification ; Sequence Alignment ; Wolbachia/*enzymology/genetics ; }, abstract = {Wolbachia pipientis are obligate endosymbionts that infect a wide range of insect and other arthropod species. They act as reproductive parasites by manipulating the host reproduction machinery to enhance their own transmission. This unusual phenotype is thought to be a consequence of the actions of secreted Wolbachia proteins that are likely to contain disulfide bonds to stabilize the protein structure. In bacteria, the introduction or isomerization of disulfide bonds in proteins is catalyzed by Dsb proteins. The Wolbachia genome encodes two proteins, alpha-DsbA1 and alpha-DsbA2, that might catalyze these steps. In this work we focussed on the 234 residue protein alpha-DsbA1; the gene was cloned and expressed in Escherichia coli, the protein was purified and its identity confirmed by mass spectrometry. The sequence identity of alpha-DsbA1 for both dithiol oxidants (E. coli DsbA, 12%) and disulfide isomerases (E. coli DsbC, 14%) is similar. We therefore sought to establish whether alpha-DsbA1 is an oxidant or an isomerase based on functional activity. The purified alpha-DsbA1 was active in an oxidoreductase assay but had little isomerase activity, indicating that alpha-DsbA1 is DsbA-like rather than DsbC-like. This work represents the first successful example of the characterization of a recombinant Wolbachia protein. Purified alpha-DsbA1 will now be used in further functional studies to identify protein substrates that could help explain the molecular basis for the unusual Wolbachia phenotypes, and in structural studies to explore its relationship to other disulfide oxidoreductase proteins.}, } @article {pmid18386064, year = {2008}, author = {Jacob, J and Mitreva, M and Vanholme, B and Gheysen, G}, title = {Exploring the transcriptome of the burrowing nematode Radopholus similis.}, journal = {Molecular genetics and genomics : MGG}, volume = {280}, number = {1}, pages = {1-17}, pmid = {18386064}, issn = {1617-4615}, support = {AI 46593/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Databases, Genetic ; *Gene Expression Profiling ; Genes, Helminth/physiology ; Host-Parasite Interactions/genetics ; Nematoda/*genetics ; RNA, Helminth/genetics/physiology ; RNA, Small Interfering/genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Wolbachia/genetics ; }, abstract = {Radopholus similis is an important nematode pest on fruit crops in the tropics. Unraveling the transcriptome of this migratory plant-parasitic nematode can provide insight in the parasitism process and lead to more efficient control measures. For the first high throughput molecular characterization of this devastating nematode, 5,853 expressed sequence tags from a mixed stage population were generated. Adding 1,154 tags from the EST division of GenBank for subsequent analysis, resulted in a total of 7,007 ESTs, which represent approximately 3,200 genes. The mean G + C content of the nucleotides at the third codon position (GC3%) was calculated to be as high as 64.8%, the highest for nematodes reported to date. BLAST-searches resulted in about 70% of the clustered ESTs having homology to (DNA and protein) sequences from the GenBank database, whereas one-third of them did not match to any known sequence. Roughly 40% of these latter sequences are predicted to be coding, representing putative novel protein coding genes. Functional annotation of the sequences by GO annotation revealed the abundance of genes involved in reproduction and development, which reflects the nematode population biology. Genes with a role in the parasitism process are identified, as well as genes essential for nematode survival, providing information useful for parasite control. No evidence was found for the presence of trans-spliced leader sequences commonly occurring in nematodes, despite the use of various approaches. In conclusion, we found three different sources for the EST sequences: the majority has a nuclear origin, approximately 1% of the EST sequences are derived from the mitochondrial transcriptome, and interestingly, 1% of the tags are with high probability derived from Wolbachia, providing the first molecular indication for the presence of this endosymbiont in a plant-parasitic nematode.}, } @article {pmid18377816, year = {2008}, author = {Klion, AD}, title = {Filarial infections in travelers and immigrants.}, journal = {Current infectious disease reports}, volume = {10}, number = {1}, pages = {50-57}, pmid = {18377816}, issn = {1523-3847}, abstract = {Filarial infections including loiasis, onchocerciasis, and lymphatic filariasis are important causes of morbidity in endemic populations worldwide, and they present a risk to travelers to endemic areas. Definitive diagnosis is complicated by overlap in the geographic distribution and clinical manifestations of the different filarial parasites, as well as similarities in their antigenic and nucleic acid composition. This has important implications for treatment, because the efficacies and toxicities of available antifilarial agents differ dramatically among filarial species. Recent advances, including the visualization of adult filarial worms in vivo by high-frequency ultrasound and the identification of the bacterial endosymbiont, Wolbachia, have greatly improved our understanding of the pathogenesis of filarial infection and have led to novel approaches to the diagnosis and treatment of travelers and immigrants from filarial-endemic regions.}, } @article {pmid18365535, year = {2007}, author = {Bonofiglio, M and Hay, J and McPherson, C}, title = {Lymphatic filariasis: inflammatory response to Wolbachia bacteria in filarial worms.}, journal = {Lymphology}, volume = {40}, number = {4}, pages = {191-192}, pmid = {18365535}, issn = {0024-7766}, mesh = {Animals ; Elephantiasis, Filarial/microbiology/*parasitology/pathology ; Filarioidea/immunology/*microbiology ; Humans ; Inflammation ; Lymphatic Vessels/*parasitology/pathology ; Symbiosis ; Wolbachia/*physiology ; }, } @article {pmid18355299, year = {2008}, author = {van Overbeek, L and Gassner, F and van der Plas, CL and Kastelein, P and Nunes-da Rocha, U and Takken, W}, title = {Diversity of Ixodes ricinus tick-associated bacterial communities from different forests.}, journal = {FEMS microbiology ecology}, volume = {66}, number = {1}, pages = {72-84}, doi = {10.1111/j.1574-6941.2008.00468.x}, pmid = {18355299}, issn = {0168-6496}, mesh = {Animals ; Bacteria/*genetics ; *Biodiversity ; Borrelia/genetics/growth & development ; DNA, Bacterial/genetics ; Genes, Bacterial ; Genes, rRNA ; Geography ; Ixodes/*microbiology ; Netherlands ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Species Specificity ; }, abstract = {Nymphal Ixodes ricinus ticks (n=180) were collected from three different areas in the Netherlands to investigate the effect of forest composition on tick-associated microbial communities. Sampled habitats differed in thickness of leaf litter and humus layers and vegetation associations and were located near Amsterdam (Beech-Oak), Ede (Birch-Oak) and Veldhoven (Birch-Oak). Analysis of nine 16S rRNA gene clone libraries made from individual ticks showed nearest matches with presumed pathogens Candidatus Neoehrlichia mikurensis and Rickettsia australis and arthropod endosymbionts Wolbachia pipientis and Candidatus Midichloria mitochondrii. Total bacterial species diversity (Shannon index) and Borrelia species infections were determined in I. ricinus by, respectively, PCR-denaturing gradient gel-electrophoresis and PCR-reverse line blot with probes specific for Borrelia burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia valaisiana, Borrelia lusitaniae and Borrelia ruski. Bacterial diversity differed significantly per area and was lowest in Ede. In contrast, Borrelia species-infected ticks were more abundant in Ede, Candidatus Neoehrlichia mikurensis-infected ticks in Ede and Veldhoven, and R. australis-infected ticks in Amsterdam. Borrelia afzelii was the most common Borrelia species found in all three areas. Bacterial tick diversity was influenced by local differences in forest structure, which is proposed to modulate animal populations that are commonly parasitized by I. ricinus.}, } @article {pmid18339571, year = {2008}, author = {Korten, S and Badusche, M and Büttner, DW and Hoerauf, A and Brattig, N and Fleischer, B}, title = {Natural death of adult Onchocerca volvulus and filaricidal effects of doxycycline induce local FOXP3+/CD4+ regulatory T cells and granzyme expression.}, journal = {Microbes and infection}, volume = {10}, number = {3}, pages = {313-324}, doi = {10.1016/j.micinf.2007.12.004}, pmid = {18339571}, issn = {1286-4579}, mesh = {Animals ; Antigens, Helminth/immunology/metabolism ; Burkina Faso ; CD4 Antigens/*metabolism ; Doxycycline/pharmacology/*therapeutic use ; Female ; Filaricides/pharmacology/*therapeutic use ; Forkhead Transcription Factors/*metabolism ; Ghana ; Granzymes/*metabolism ; Humans ; Immunohistochemistry ; Liberia ; *Onchocerca volvulus/drug effects ; Onchocerciasis/*drug therapy/*immunology ; Random Allocation ; T-Lymphocytes, Regulatory/*immunology/*metabolism ; Treatment Outcome ; }, abstract = {Immunosuppression in human filarial disease involves regulatory T cells. We hypothesized that natural or worm antigen-induced FOXP3 regulatory T cells could be involved locally, suppressing effector cells via granzymes. Natural and treatment-induced death of worms implies enhanced exposure to worm antigens. Therefore, we examined FOXP3+T cells and granzyme expression in onchocercomas harbouring adult Onchocerca volvulus worms, with respect to worm viability, productivity, the patient's immune status and filaricidal treatment. The immunohistological analysis revealed that dead adult worms were strongly associated with FOXP3+T cells in generalized hyporeactive onchocerciasis. FOXP3+ cells hardly expressed granzymes, but cell contacts with granzyme A+ or B+ cells were frequent. While suramin directly kills most adult worms within 6 months, the Wolbachia depleting antibiotic doxycycline indirectly causes adult worm degeneration within 18 months. Contrary to suramin, depletion of Th1-driving endobacteria most strongly promoted FOXP3+T cells and granzyme-expressing cells. In hyperreactive patients, FOXP3+ cells were less frequent. This is the first demonstration of local FOXP3+Treg cells in human filariasis and their induction by natural worm death and anti-parasitic treatment. We newly report granzyme responses to helminths and their association with immunosuppression. FOXP3+Treg and granzyme+ cells might locally suppress defence against newly acquired worms.}, } @article {pmid18338578, year = {2008}, author = {Guo, X and Li, Z}, title = {[Wolbachia extensively harbored by Bemisia tabaci in China].}, journal = {Wei sheng wu xue bao = Acta microbiologica Sinica}, volume = {48}, number = {1}, pages = {63-67}, pmid = {18338578}, issn = {0001-6209}, mesh = {Animals ; China ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Hemiptera/genetics/*microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia are a group of maternally inherited bacteria harbored by a variety of arthropods and can manipulate the reproductivity of their hosts. Eighteen populations of whiteflies were collected from Hebei, Xinjiang, Beijing, Shandong, Zhejiang, Guangxi, Hainan, Guangzhou and Fujian provinces, China. These whiteflies were molecularly identified using internal transcribed spacer 1 ITS1 rDNA sequences. They were then detected for Wolbachia infection using Wolbachia-specific primers designed based on 16S rDNA and wsp gene sequences. The results showed that almost all populations were detected positive for Wolbachia infection. Whiteflies of B/Q group mainly carried Wolbachia belonging to supergroup A, while non- B/Q whiteflies were commonly detected for Wolbachia superinfection. This study indicated that the infection rate of Wolbachia in natural populations of B. tabaci might be much higher than expected, and the threshold of detection methods may be one of the key factors influencing detection of Wolbachia infection.}, } @article {pmid18329099, year = {2008}, author = {Herbinière, J and Grève, P and Strub, JM and Thiersé, D and Raimond, M and van Dorsselaer, A and Martin, G and Braquart-Varnier, C}, title = {Protein profiling of hemocytes from the terrestrial crustacean Armadillidium vulgare.}, journal = {Developmental and comparative immunology}, volume = {32}, number = {8}, pages = {875-882}, doi = {10.1016/j.dci.2008.01.007}, pmid = {18329099}, issn = {0145-305X}, mesh = {Animals ; Base Sequence ; Cell Communication ; Crustacea/chemistry/*immunology ; Cytoskeletal Proteins/analysis ; DNA, Complementary/isolation & purification ; Hemocytes/*chemistry ; Immunity, Innate ; Melanins/biosynthesis ; Molecular Sequence Data ; *Proteomics ; Superoxide Dismutase/analysis ; Transglutaminases/analysis ; }, abstract = {To establish and maintain a successful infection, microbial pathogens have evolved various strategies to infect the host in the face of a functional immune system. In this context, the alpha-proteobacteria Wolbachia capacities to infect new host species have been greatly evidenced. Indeed, in terrestrial isopods, experimentally transferred Wolbachia invade all host tissues, including immune cells such as hemocytes. To investigate mechanisms that have to be avoided by bacteria to maintain themselves in hemocytes, we characterized the hemocyte proteome of Armadillidium vulgare by a 2D gel electrophoresis approach. Fifty-six proteins were identified and classified into functional groups (stress and immunity, glucose metabolisms, cytoskeleton, others). We focused on immune response and cytoskeleton proteins often exploited by bacteria to invade their host. From the microsequences obtained by mass spectrometry, PCR primers were designed to amplify seven partial cDNAs encoding masquerade, alpha2-macroglobulin, transglutaminase, MnSOD, calreticulin, cyclophilin, and vinculin, confirming their expression in hemocytes.}, } @article {pmid18312577, year = {2008}, author = {Hilgenboecker, K and Hammerstein, P and Schlattmann, P and Telschow, A and Werren, JH}, title = {How many species are infected with Wolbachia?--A statistical analysis of current data.}, journal = {FEMS microbiology letters}, volume = {281}, number = {2}, pages = {215-220}, pmid = {18312577}, issn = {0378-1097}, mesh = {Animals ; Arthropods/classification/*microbiology ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia are intracellular bacteria found in many species of arthropods and nematodes. They manipulate the reproduction of their arthropod hosts in various ways, may play a role in host speciation and have potential applications in biological pest control. Estimates suggest that at least 20% of all insect species are infected with Wolbachia. These estimates result from several Wolbachia screenings in which numerous species were tested for infection; however, tests were mostly performed on only one to two individuals per species. The actual percent of species infected will depend on the distribution of infection frequencies among species. We present a meta-analysis that estimates percentage of infected species based on data on the distribution of infection levels among species. We used a beta-binomial model that describes the distribution of infection frequencies of Wolbachia, shedding light on the overall infection rate as well as on the infection frequency within species. Our main findings are that (1) the proportion of Wolbachia-infected species is estimated to be 66%, and that (2) within species the infection frequency follows a 'most-or-few' infection pattern in a sense that the Wolbachia infection frequency within one species is typically either very high (>90%) or very low (<10%).}, } @article {pmid18302997, year = {2008}, author = {Nakanishi, K and Hoshino, M and Nakai, M and Kunimi, Y}, title = {Novel RNA sequences associated with late male killing in Homona magnanima.}, journal = {Proceedings. Biological sciences}, volume = {275}, number = {1640}, pages = {1249-1254}, pmid = {18302997}, issn = {0962-8452}, mesh = {Animals ; Base Sequence ; Female ; *Genes, Insect ; *Genes, Lethal ; Male ; Molecular Sequence Data ; Moths/*genetics/*growth & development ; RNA/*genetics/isolation & purification ; Sex Ratio ; Species Specificity ; }, abstract = {Maternally inherited female-biased sex ratios have been documented in many invertebrate species. One cause of such biased sex ratios is male killing, i.e. only males die. In most species, male killing occurs during embryonic stages (early male killing) and is associated with cytoplasmic bacteria, including Wolbachia, Spiroplasma, Rickettsia, Flavobacteria and gamma proteobacteria. However, the oriental tea tortrix, Homona magnanima, is one of the few species in which male death occurs in the larval or pupal stage, and is thus an example of late male killing. We partially purified the agent causing late male killing in H. magnanima and showed that it consists of two RNA sequences. This represents an entirely novel agent causing late male killing.}, } @article {pmid18302730, year = {2008}, author = {Fuxelius, HH and Darby, AC and Cho, NH and Andersson, SG}, title = {Visualization of pseudogenes in intracellular bacteria reveals the different tracks to gene destruction.}, journal = {Genome biology}, volume = {9}, number = {2}, pages = {R42}, pmid = {18302730}, issn = {1474-760X}, mesh = {Base Sequence ; Evolution, Molecular ; *Genes, Bacterial ; *Genetic Variation ; Interspersed Repetitive Sequences ; Open Reading Frames/genetics ; Phylogeny ; Proteobacteria/genetics ; *Pseudogenes ; Rickettsia/*genetics ; Sequence Analysis, DNA/*methods ; *Software ; }, abstract = {BACKGROUND: Pseudogenes reveal ancestral gene functions. Some obligate intracellular bacteria, such as Mycobacterium leprae and Rickettsia spp., carry substantial fractions of pseudogenes. Until recently, horizontal gene transfers were considered to be rare events in obligate host-associated bacteria.

RESULTS: We present a visualization tool that displays the relationships and positions of degraded and partially overlapping gene sequences in multiple genomes. With this tool we explore the origin and deterioration patterns of the Rickettsia pseudogenes and find that variably present genes and pseudogenes tend to have been acquired more recently, are more divergent in sequence, and exhibit a different functional profile compared with genes conserved across all species. Overall, the origin of only one-quarter of the variable genes and pseudogenes can be traced back to the common ancestor of Rickettsia and the outgroup genera Orientia and Wolbachia. These sequences contain only a few disruptive mutations and show a broad functional distribution profile, much like the core genes. The remaining genes and pseudogenes are extensively degraded or solely present in a single species. Their functional profile was heavily biased toward the mobile gene pool and genes for components of the cell wall and the lipopolysaccharide.

CONCLUSION: Reductive evolution of the vertically inherited genomic core accounts for 25% of the predicted genes in the variable segments of the Rickettsia genomes, whereas 75% stems from the flux of the mobile gene pool along with genes for cell surface structures. Thus, most of the variably present genes and pseudogenes in Rickettsia have arisen from recent acquisitions.}, } @article {pmid18298644, year = {2008}, author = {Hornett, EA and Duplouy, AM and Davies, N and Roderick, GK and Wedell, N and Hurst, GD and Charlat, S}, title = {You can't keep a good parasite down: evolution of a male-killer suppressor uncovers cytoplasmic incompatibility.}, journal = {Evolution; international journal of organic evolution}, volume = {62}, number = {5}, pages = {1258-1263}, doi = {10.1111/j.1558-5646.2008.00353.x}, pmid = {18298644}, issn = {0014-3820}, support = {//Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; *Biological Evolution ; Breeding ; Butterflies/cytology/*microbiology ; Cytoplasm/metabolism ; Female ; Host-Parasite Interactions ; Male ; Phenotype ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Maternally inherited parasites are known to impose a wide variety of reproductive manipulations upon their host. These often produce strong selection on the host to suppress the parasite, resulting in a reduction in the frequency of the parasite. However, in the butterfly Hypolimnas bolina, infected with a Wolbachia bacterium, field data demonstrate that suppression of the male-killing phenotype does not depress parasite frequency. Here we test and verify one hypothesis to explain this apparent paradox-Wolbachia induces a second phenotype, Cytoplasmic Incompatibility (CI), in populations where host suppression has evolved. We further demonstrate that the capacity to induce CI has not evolved de novo, but instead is instantaneously expressed upon the survival of infected males. The significance of these results is threefold: (1) multiple phenotypes can provide Wolbachia with the means to maintain itself in a host following suppression of a single manipulative phenotype; (2) the ability to induce CI can remain hidden in systems in which male-killing is observed, just as the ability to induce male-killing may be obscured in strains exhibiting CI; (3) the evolutionary maintenance of CI in a system in which it is not expressed suggests a functional link with male-killing or other traits under selection.}, } @article {pmid18291041, year = {2008}, author = {Viljakainen, L and Reuter, M and Pamilo, P}, title = {Wolbachia transmission dynamics in Formica wood ants.}, journal = {BMC evolutionary biology}, volume = {8}, number = {}, pages = {55}, pmid = {18291041}, issn = {1471-2148}, mesh = {Animals ; Ants/classification/*microbiology ; Bacterial Outer Membrane Proteins/genetics ; DNA, Bacterial/genetics ; DNA, Mitochondrial/genetics ; Evolution, Molecular ; *Gene Transfer, Horizontal ; Genetic Variation ; Haplotypes ; *Host-Pathogen Interactions ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Wolbachia/classification/*physiology ; }, abstract = {BACKGROUND: The role of Wolbachia endosymbionts in shaping the mitochondrial diversity of their arthropod host depends on the effects they have on host reproduction and on the mode of transmission of the bacteria. We have compared the sequence diversity of wsp (Wolbachia surface protein gene) and the host mtDNA in a group of Formica ant species that have diverged approximately 0.5 million years ago (MYA). The aim was to study the relationship of Wolbachia and its ant hosts in terms of vertical and horizontal transmission of the bacteria.

RESULTS: All studied ant species were doubly infected with two Wolbachia strains (wFex1 and wFex4) all over their geographical distribution area in Eurasia. The most common haplotypes of these strains were identical with strains previously described from a more distantly related Formica ant, with an estimated divergence time of 3.5 - 4 MYA. Some strain haplotypes were associated to the same or closely related mtDNA haplotypes as expected under vertical transmission. However, in several cases the wsp haplotypes coexisted with distant mtDNA haplotypes, a pattern which is more compatible with horizontal transmission of the bacteria.

CONCLUSION: Two lines of evidence suggest that the sharing of Wolbachia strains by all F. rufa species is rather due to horizontal than vertical transmission. First, the fact that endosymbiont strains identical to those of F. rufa ants have been found in another species that diverged 3.5-4 MYA strongly suggests that horizontal transfer can and does occur between Formica ants. Second, the frequent sharing of identical Wolbachia strains by distant mitochondrial lineages within the F. rufa group further shows that horizontal transmission has occurred repeatedly. Nevertheless, our dataset also provides some evidence for longer-term persistence of infection, indicating that Wolbachia infection within this host clade has been shaped by both horizontal and vertical transmission of symbionts. The fact that all the ants were infected irrespective of the family structure of their societies gives no support to the proposed hypotheses that the spreading of Wolbachia in ants might be associated to the types of their societies.}, } @article {pmid18289290, year = {2008}, author = {Levy, JK and Crawford, PC and Lappin, MR and Dubovi, EJ and Levy, MG and Alleman, R and Tucker, SJ and Clifford, EL}, title = {Infectious diseases of dogs and cats on Isabela Island, Galapagos.}, journal = {Journal of veterinary internal medicine}, volume = {22}, number = {1}, pages = {60-65}, pmid = {18289290}, issn = {0891-6640}, mesh = {Animals ; Cat Diseases/*epidemiology ; Cats ; Communicable Diseases/epidemiology/*veterinary ; Cross-Sectional Studies ; Dog Diseases/*epidemiology ; Dogs ; Ecuador/epidemiology ; Endemic Diseases/veterinary ; Female ; Male ; Prevalence ; Prospective Studies ; }, abstract = {BACKGROUND: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans.

HYPOTHESIS: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations.

ANIMALS: Ninety-five dogs and 52 cats presented during a neutering campaign.

METHODS: A prospective cross-sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases.

RESULTS: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66-67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia/Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis.

Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions.}, } @article {pmid18285399, year = {2008}, author = {Gottlieb, Y and Ghanim, M and Gueguen, G and Kontsedalov, S and Vavre, F and Fleury, F and Zchori-Fein, E}, title = {Inherited intracellular ecosystem: symbiotic bacteria share bacteriocytes in whiteflies.}, journal = {FASEB journal : official publication of the Federation of American Societies for Experimental Biology}, volume = {22}, number = {7}, pages = {2591-2599}, doi = {10.1096/fj.07-101162}, pmid = {18285399}, issn = {1530-6860}, mesh = {Animals ; *Bacterial Physiological Phenomena ; *Ecosystem ; Female ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Intracellular Space/*physiology ; Ovary/microbiology ; Ovum/microbiology ; Rickettsia/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Symbiotic relationships with bacteria are common within the Arthropoda, with interactions that substantially influence the biology of both partners. The symbionts' spatial distribution is essential for understanding key aspects of this relationship, such as bacterial transmission, phenotype, and dynamics. In this study, fluorescence in situ hybridization was used to localize five secondary symbionts from various populations and biotypes of the sweet potato whitefly Bemisia tabaci: Hamiltonella, Arsenophonus, Cardinium, Wolbachia, and Rickettsia. All five symbionts were found to be located with the primary symbiont Portiera inside the bacteriocytes--cells specifically modified to house bacteria--but within these cells, they occupied various niches. The intrabacteriocyte distribution pattern of Rickettsia differed from what has been described previously. Cardinium and Wolbachia were found in other host tissues as well. Because all symbionts share the same cell, bacteriocytes in B. tabaci represent a unique intracellular ecosystem. This phenomenon may be a result of the direct enclosure of the bacteriocyte in the egg during oogenesis, providing a useful mechanism for efficient vertical transmission by "hitching a ride" with Portiera. On the other hand, cohabitation in the same cell provides ample opportunities for interactions among symbionts that can either facilitate (cooperation) or limit (warfare) symbiotic existence.}, } @article {pmid18284754, year = {2007}, author = {Sun, X and Cui, L and Li, Z}, title = {Diversity and phylogeny of Wolbachia infecting Bactrocera dorsalis (Diptera: Tephritidae) populations from China.}, journal = {Environmental entomology}, volume = {36}, number = {5}, pages = {1283-1289}, doi = {10.1603/0046-225x(2007)36[1283:dapowi]2.0.co;2}, pmid = {18284754}, issn = {0046-225X}, mesh = {Animals ; China ; *DNA, Mitochondrial ; Electron Transport Complex IV/*genetics ; Genetic Variation ; Geography ; *Phylogeny ; Sequence Analysis, DNA ; Tephritidae/genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia are a common and widespread group of symbiotic bacteria found in the reproductive tissues of arthropods. Bactrocera dorsalis (Hendel) is an important pest causing considerable economic losses of fruits and vegetables in several southern provinces of China. In this study, polymerase chain reaction (PCR) with general Wolbachia surface protein (wsp) primers was used to test the presence of Wolbachia in 1,500 individuals of B. dorsalis from five geographical populations of China. We detected 19 individuals of B. dorsalis infected by Wolbachia, and the infection rates of different populations varied. Comparison of wsp gene sequences from 19 individuals and search of the GenBank identified four new sequences, probably representing four Wolbachia strains. Sequence comparison showed that the four Wolbachia strains from B. dorsalis in China belonged to three groups (Kue, Mel, and Cuc). Phylogenetic analysis of the wsp sequences suggests that geographical isolation of Wolbachia exists among the populations of B. dorsalis in China, and gene flow of Wolbachia might have occurred between B. dorsalis populations of China and Thailand. Phylogenetic analysis performed on the host mitochondrial cytochrome oxidase I (COI) gene and wsp gene suggests that host has coevolved with Wolbachia.}, } @article {pmid18282572, year = {2008}, author = {Arumugam, S and Pfarr, KM and Hoerauf, A}, title = {Infection of the intermediate mite host with Wolbachia-depleted Litomosoides sigmodontis microfilariae: impaired L1 to L3 development and subsequent sex-ratio distortion in adult worms.}, journal = {International journal for parasitology}, volume = {38}, number = {8-9}, pages = {981-987}, doi = {10.1016/j.ijpara.2007.12.006}, pmid = {18282572}, issn = {0020-7519}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/*pharmacology ; DNA, Bacterial/analysis ; DNA, Helminth/analysis/drug effects ; Gerbillinae/*parasitology ; Humans ; Male ; Microfilariae/*growth & development/microbiology ; Mites/parasitology ; Polymerase Chain Reaction ; *Sex Ratio ; Tetracycline/administration & dosage/*pharmacology ; Wolbachia/*drug effects/isolation & purification ; }, abstract = {The rodent filaria Litomosoides sigmodontis harbour Wolbachia, endosymbionts essential for worm embryogenesis, larval development and adult survival. To study the effect of tetracycline, which depletes Wolbachia, on the development of microfilariae (L1s, MF) to L3 in the intermediate host Ornithonyssus bacoti, and to observe the development of Wolbachia-depleted L3s in Mongolian gerbils (Meriones unguiculatus); microfilaremic gerbils were treated orally with tetracycline for 6 weeks (primary infected Tet) or untreated (primary Con). Treatment resulted in a significant reduction of Wolbachia per MF in primary Tet gerbils. Naïve mites then fed on the primary Tet and primary Con gerbils in the week after treatment ended, when MF levels were not significantly different, and used to infect new gerbils (secondary infected) Tet, secondary Con) via natural infection. The infection rate from dissected mites was 9% and 54% (primary Tet and primary Con, respectively). After 3 months, worms were isolated from secondary gerbils. Significantly fewer female worms developed in secondary Tet gerbils. In contrast, there was no difference in the number of male worms that developed in secondary gerbils, resulting in a male biased sex-ratio. Although secondary Tet male worms had fewer Wolbachia than secondary Con males, development was not impaired. Female worms that developed from Wolbachia-depleted MF had Wolbachia levels equivalent to worms from secondary Con animals. Thus, tetracycline pre-treatment selected for female worms with high numbers of Wolbachia, whereas male worms had median Wolbachia levels significantly lower than secondary Con males. Therefore, female worms require a higher threshold of Wolbachia for their development. The worms analysed were only exposed to tetracycline as MF, ruling out direct effects of tetracycline during larval development in the mites or secondary gerbils, suggesting that the depletion of Wolbachia in MF was the cause of impaired larval development.}, } @article {pmid18266629, year = {2008}, author = {Duron, O and Hurst, GD and Hornett, EA and Josling, JA and Engelstädter, J}, title = {High incidence of the maternally inherited bacterium Cardinium in spiders.}, journal = {Molecular ecology}, volume = {17}, number = {6}, pages = {1427-1437}, doi = {10.1111/j.1365-294X.2008.03689.x}, pmid = {18266629}, issn = {1365-294X}, mesh = {Animals ; Bacterial Infections/microbiology ; Bacteroidetes/*physiology ; *Biological Evolution ; DNA, Ribosomal/genetics ; Female ; Inheritance Patterns/*genetics ; Male ; Mitochondria/genetics ; Phylogeny ; Sex Ratio ; Spiders/genetics/*microbiology/*physiology ; Wolbachia/physiology ; }, abstract = {Inherited bacteria are now recognized as important players in arthropod evolution and ecology. Here, we test spiders, a group recently identified as possessing inherited bacteria commonly, for the presence of two reproductive parasites, Cardinium hertigii (Bacteroidetes group) and Wolbachia (alpha-proteobacteria), estimating incidence, prevalence, any sex bias in infection, and infection diversity, for a panel of field-collected specimens. We identify spiders as a hotspot for Cardinium. Present in 22% of the sampled species, incidence was significantly higher than that previously recorded in insects. Where present, Cardinium infection occurred at medium prevalence without evidence of sex bias in prevalence that would indicate sex-ratio distortion activity. Wolbachia was present in 37% of species, but revealed a gradation from being rare to very common. In one case, Wolbachia was found significantly more commonly in females than males, indicating it may act as a sex-ratio distorter in some species. Breeding work conducted on two species confirmed that Wolbachia and Cardinium were transmitted maternally, which represents the first proof of inheritance of these symbionts in spiders. Overall, this study demonstrates that the majority of spider species are infected with inherited bacteria, and their role in host biology clearly requires determination.}, } @article {pmid18259058, year = {2008}, author = {Kurz, M and Iturbe-Ormaetxe, I and Jarrott, R and O'Neill, SL and Byriel, KA and Martin, JL and Heras, B}, title = {Crystallization and preliminary diffraction analysis of a DsbA homologue from Wolbachia pipientis.}, journal = {Acta crystallographica. Section F, Structural biology and crystallization communications}, volume = {64}, number = {Pt 2}, pages = {94-97}, pmid = {18259058}, issn = {1744-3091}, mesh = {Crystallization ; Crystallography, X-Ray ; Polymerase Chain Reaction ; Protein Disulfide-Isomerases/*chemistry ; Recombinant Proteins/chemistry ; Wolbachia/*chemistry ; }, abstract = {alpha-DsbA1 is one of two DsbA homologues encoded by the Gram-negative alpha-proteobacterium Wolbachia pipientis, an endosymbiont that can behave as a reproductive parasite in insects and as a mutualist in medically important filarial nematodes. The alpha-DsbA1 protein is thought to be important for the folding and secretion of Wolbachia proteins involved in the induction of reproductive distortions. Crystals of native and SeMet alpha-DsbA1 were grown by vapour diffusion and belong to the monoclinic space group C2, with unit-cell parameters a = 71.4, b = 49.5, c = 69.3 A, beta = 107.0 degrees and one molecule in the asymmetric unit (44% solvent content). X-ray data were recorded from native crystals to a resolution of 2.01 A using a copper anode and data from SeMet alpha-DsbA1 crystals were recorded to 2.45 A resolution using a chromium anode.}, } @article {pmid18257289, year = {2007}, author = {Nefedova, LN and Kim, AI}, title = {[Evolution from retrotransposons to retroviruses: origin of the env gene].}, journal = {Zhurnal obshchei biologii}, volume = {68}, number = {6}, pages = {459-467}, pmid = {18257289}, issn = {0044-4596}, mesh = {Animals ; Drosophila melanogaster ; *Evolution, Molecular ; Gene Products, env/*genetics ; Genome, Insect/physiology ; Retroelements/*physiology ; Retroviridae/*physiology ; }, abstract = {In genome of Drosophila melanogaster, various families of retrotransposons with different combination of functional domens and mechanisms of transposition are present. However only retrotransposons of gypsy family are retroviruses related to errantiviruses. Other families seemingly appeared as intermediate forms of retroviruses evolution. Despite the fact that the question on origin of retroviruses remains unclear, now the hypothesis of their origin from retrotransoposons can be considered the most consistent. Infectious properties of errantiviruses are linked to the presence of the third open reading frame (the env gene). Acquisition of the env gene conversed retrotransposons into retroviruses. So, origin of this gene is of special interest. Homologues of the env gene of errantiviruses are discovered in genomes of D. melanogaster, as well as in baculoviruses and in bacteria Wolbachia pipientis, the endosymbiont of Drosophila. It was shown that homologue of the env gene come to Wolbachia genome from Drosophila genome by horizontal transfer of the gypsy group retrotransposon. Thus, Wolbachia was not a donor of the env gene for errantiviruses. Seemingly, errantiviruses captured the baculoviral homologue of the env gene (f). However origin of the f gene is not clear. At the same time the env gene homologue in D. melanogaster genome exist (Iris). It must not be ruled out that the Iris gene was the source of the env gene of errantiviruses and baculoviruses.}, } @article {pmid18245338, year = {2008}, author = {Perlman, SJ and Kelly, SE and Hunter, MS}, title = {Population biology of cytoplasmic incompatibility: maintenance and spread of Cardinium symbionts in a parasitic wasp.}, journal = {Genetics}, volume = {178}, number = {2}, pages = {1003-1011}, pmid = {18245338}, issn = {0016-6731}, mesh = {Animals ; Bacteroidetes/*physiology ; Crosses, Genetic ; Cytoplasm/microbiology ; Female ; Fertility ; Male ; Oviposition ; Pest Control, Biological ; Reproduction ; Solanum tuberosum/parasitology ; Symbiosis/*physiology ; Wasps/*microbiology/parasitology/*physiology ; }, abstract = {Bacteria that cause cytoplasmic incompatibility (CI) are perhaps the most widespread parasites of arthropods. CI symbionts cause reproductive failure when infected males mate with females that are either uninfected or infected with a different, incompatible strain. Until recently, CI was known to be caused only by the alpha-proteobacterium Wolbachia. Here we present the first study of the population biology of Cardinium, a recently discovered symbiont in the Bacteroidetes that causes CI in the parasitic wasp Encarsia pergandiella (Hymenoptera: Aphelinidae). Cardinium occurs at high frequency (approximately 92%) in the field. Using wasps that were recently collected in the field, we measured parameters that are crucial for understanding how CI spreads and is maintained in its host. CI Cardinium exhibits near-perfect rates of maternal transmission, causes a strong reduction in viable offspring in incompatible crosses, and induces a high fecundity cost, with infected females producing 18% fewer offspring in the first 4 days of reproduction. We found no evidence for paternal transmission or horizontal transmission of CI Cardinium through parasitism of an infected conspecific. No evidence for cryptic parthenogenesis in infected females was found, nor was sex allocation influenced by infection. We incorporated our laboratory estimates into a model of CI dynamics. The model predicts a high stable equilibrium, similar to what we observed in the field. Interestingly, our model also predicts a high threshold frequency of CI invasion (20% for males and 24% for females), below which the infection is expected to be lost. We consider how this threshold may be overcome, focusing in particular on the sensitivity of CI models to fecundity costs. Overall our results suggest that the factors governing the dynamics of CI Wolbachia and Cardinium are strikingly similar.}, } @article {pmid18236077, year = {2008}, author = {Cárdenas, MQ and Lanfredi, RM}, title = {Ultrastructure of reproductive system of female Litomosoides chagasfilhoi Moraes-Neto, Lanfredi and De Souza, 1997 (Nematoda: Filarioidea).}, journal = {Parasitology research}, volume = {102}, number = {6}, pages = {1135-1142}, pmid = {18236077}, issn = {0932-0113}, mesh = {Animals ; Cytoplasm/microbiology/ultrastructure ; Epithelial Cells/ultrastructure ; Female ; Filarioidea/*ultrastructure ; Genitalia, Female/ultrastructure ; Male ; Microscopy, Electron, Transmission ; Muscle Cells/ultrastructure ; Oocytes/ultrastructure ; Spermatozoa/microbiology/ultrastructure ; }, abstract = {The ultrastructure of the female reproductive system of the filariid Litomosoides chagasfilhoi by transmission electron microscopy (TEM) is described for the first time. The ovary is composed of primary oocytes surrounded by a single layer of epithelial cells apposed on the basal laminae. The ovarian wall is completely filled with primary oocytes, which are arranged radially and are centrally connected around the rachis. The uterine wall consists of muscular fibers surrounded by a basal lamina and the epithelium underlying the lamina. Ameboid and aflagellate spermatozoa are present inside the distal portion of the uterus, some of them near oocytes, which present bacteria in its cytoplasm. An eletrondense well-defined eggshell covers the zygotes, which presents in its cytoplasm bacteria arranged in groups. These bacteria are also observed in embryos and in the hypodermal cord. These ultrastructural aspects of L. chagasfilhoi female worms presented herein contribute to the knowledge of the morphology and embryonary development of this filariid, providing means for further comparative analyses of the action of anti-filarial drugs. Besides this, the presence of bacteria Wolbachia-like is being reported for the first time in this species, showing the great importance of this experimental model of study.}, } @article {pmid18235849, year = {2008}, author = {Brelsfoard, CL and Séchan, Y and Dobson, SL}, title = {Interspecific hybridization yields strategy for South Pacific filariasis vector elimination.}, journal = {PLoS neglected tropical diseases}, volume = {2}, number = {1}, pages = {e129}, pmid = {18235849}, issn = {1935-2735}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; R01 AI067434/AI/NIAID NIH HHS/United States ; R01-AI067434/AI/NIAID NIH HHS/United States ; R01-AI051533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/growth & development/*microbiology ; Animals ; *Chimera ; Elephantiasis, Filarial/prevention & control/*transmission ; Female ; Insect Vectors/growth & development/*microbiology ; Male ; Mosquito Control/*methods ; Wolbachia/growth & development/*physiology ; }, abstract = {BACKGROUND: Lymphatic filariasis (LF) is a leading cause of disability in South Pacific regions, where >96% of the 1.7 million population are at risk of LF infection. As part of current global campaign, mass drug administration (MDA) has effectively reduced lymphatic filiariasis prevalence, but mosquito vector biology can complicate the MDA strategy. In some regions, there is evidence that the goal of LF elimination cannot be attained via MDA alone. Obligate vector mosquitoes provide additional targets for breaking the LF transmission cycle, but existing methods are ineffective for controlling the primary vector throughout much of the South Pacific, Aedes polynesiensis.

Here we demonstrate that interspecific hybridization and introgression results in an A. polynesiensis strain ('CP' strain) that is stably infected with the endosymbiotic Wolbachia bacteria from Aedes riversi. The CP strain is bi-directionally incompatible with naturally infected mosquitoes, resulting in female sterility. Laboratory assays demonstrate that CP males are equally competitive, resulting in population elimination when CP males are introduced into wild type A. polynesiensis populations.

CONCLUSIONS/SIGNIFICANCE: The findings demonstrate strategy feasibility and encourage field tests of the vector elimination strategy as a supplement to ongoing MDA efforts.}, } @article {pmid18205985, year = {2008}, author = {Pfarr, KM and Heider, U and Schmetz, C and Büttner, DW and Hoerauf, A}, title = {The mitochondrial heat shock protein 60 (HSP60) is up-regulated in Onchocerca volvulus after the depletion of Wolbachia.}, journal = {Parasitology}, volume = {135}, number = {4}, pages = {529-538}, doi = {10.1017/S003118200700409X}, pmid = {18205985}, issn = {1469-8161}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Chaperonin 60/*biosynthesis/genetics ; Doxycycline/therapeutic use ; Female ; Humans ; Immunohistochemistry ; Ivermectin/therapeutic use ; Male ; Microscopy, Electron ; Mitochondria/*metabolism ; Onchocerca volvulus/genetics/*metabolism/*microbiology ; Onchocerciasis/drug therapy/*microbiology/parasitology ; RNA, Helminth/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Symbiosis ; Transcription, Genetic ; Up-Regulation ; Wolbachia/drug effects/isolation & purification/*metabolism ; }, abstract = {Wolbachia, a genus of endosymbiotic bacteria of filarial worms, represent novel targets for anti-filarial therapy. The efficacy of compounds against Wolbachia has been evaluated using antiserum raised against the 60 kDa heat shock protein (HSP60) which binds specifically to this protein in both Wolbachia and mitochondria. It has been shown that Wolbachia stains (using such specific probes) stronger than the mitochondria in untreated Onchocerca volvulus, whereas after the depletion of Wolbachia (with drugs) staining of the mitochondria is increased. Herein, immunogold electron microscopy showed that specific anti-HSP60 serum specifically labelled Wolbachia and filarial mitochondria, and that both have distinct localization patterns, thus allowing them to be differentiated. Immunohistochemistry of O. volvulus showed that HSP60 staining is increased in the mitochondria after Wolbachia depletion in the hypodermis, epithelia, muscles, oocytes, embryos, and developing spermatozoa. This could have been the result of the antiserum preferentially binding to the Wolbachia when they are present or due to increased expression of the protein in the absence of the bacteria. To address this, mRNA levels of filarial hsp60 in O. volvulus were measured. After the depletion of Wolbachia, the transcription of hsp60 was significantly greater (7.7 fold) compared with untreated worms. We hypothesize that the increased expression of HSP60 in the absence of Wolbachia is due to a disruption of the homeostasis of the endosymbiosis.}, } @article {pmid18180957, year = {2008}, author = {Shakya, S and Bajpai, P and Sharma, S and Misra-Bhattacharya, S}, title = {Prior killing of intracellular bacteria Wolbachia reduces inflammatory reactions and improves antifilarial efficacy of diethylcarbamazine in rodent model of Brugia malayi.}, journal = {Parasitology research}, volume = {102}, number = {5}, pages = {963-972}, pmid = {18180957}, issn = {0932-0113}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/pharmacology/*therapeutic use ; *Brugia malayi/drug effects/microbiology ; Diethylcarbamazine/administration & dosage/*therapeutic use ; Disease Models, Animal ; Drug Administration Schedule ; Drug Therapy, Combination ; Female ; Filariasis/*drug therapy/immunology/parasitology ; Filaricides/administration & dosage/*therapeutic use ; Gerbillinae ; Host-Parasite Interactions ; Humans ; Inflammation/drug therapy/immunology/parasitology ; Male ; Murinae ; Tetracycline/administration & dosage/pharmacology/*therapeutic use ; Treatment Outcome ; Wolbachia/*drug effects ; }, abstract = {The discovery of the endosymbiont Wolbachia, which has a mutualistic relationship with filarial nematodes, and its importance in filarial parasite biology has provided a lead for developing novel chemotherapeutic agents against human filariasis. Wolbachia also appears to be involved in immunopathological responses as well as adverse reactions after antifilarial therapy. The aim of the present study was to explore the potential of administering anti-Wolbachial therapy before antifilarial treatment to improve the filaricidal efficacy of the present-day filaricide diethylcarbamazine. An additional objective was to minimize host inflammatory reactions using a rodent model Mastomys coucha and Meriones unguiculatus infected with human lymphatic filariid Brugia malayi. We observed: (1) a 40-day treatment schedule of tetracycline alone resulted in delayed reduction in microfilaraemia and a low degree of macrofilaricidal efficacy; (2) tetracycline therapy followed by 100 mg/kg diethylcarbamazine (DEC) x5 days led to marked reduction in microfilaraemia from day 48 onward after initiation of treatment. The combination treatment also brought about approximately 70% death of adult B. malayi and sterilization of 82.3% of the surviving female worms, thus exhibiting remarkable enhancement in the antifilarial activity of DEC; (3) tissue inflammatory reactions and pathogenesis were significantly reduced as observed by histopathology, and peritoneal macrophage mediated oxidative burst shown by fluorescence-activated cell sorting (FACS) analysis using dichlorofluorescein diacetate (DCF-DA); and (4) the characteristic filarial antigen-specific and mitogen-specific cellular unresponsiveness was significantly reversed, possibly due to marked clearance of microfilaraemia. It is therefore advisable to give an anti-Wolbachial antibiotic trial before starting antifilarial therapy to achieve maximum benefits.}, } @article {pmid18179432, year = {2008}, author = {Baldo, L and Ayoub, NA and Hayashi, CY and Russell, JA and Stahlhut, JK and Werren, JH}, title = {Insight into the routes of Wolbachia invasion: high levels of horizontal transfer in the spider genus Agelenopsis revealed by Wolbachia strain and mitochondrial DNA diversity.}, journal = {Molecular ecology}, volume = {17}, number = {2}, pages = {557-569}, doi = {10.1111/j.1365-294X.2007.03608.x}, pmid = {18179432}, issn = {0962-1083}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Gene Transfer, Horizontal/*genetics ; Genetic Variation ; Geography ; Haplotypes ; Host-Pathogen Interactions ; Phylogeny ; Spiders/classification/*genetics/microbiology ; United States ; Wolbachia/*genetics ; }, abstract = {The pandemic distribution of Wolbachia (alpha-proteobacteria) across arthropods is largely due to the ability of these maternally inherited endosymbionts to successfully shift hosts across species boundaries. Yet it remains unclear whether Wolbachia has preferential routes of transfer among species. Here, we examined populations of eight species of the North American funnel-web spider genus Agelenopsis to evaluate whether Wolbachia show evidence for host specificity and the relative contribution of horizontal vs. vertical transmission of strains within and among related host species. Wolbachia strains were characterized by multilocus sequence typing (MLST) and Wolbachia surface protein (WSP) sequences, and analysed in relation to host phylogeny, mitochondrial diversity and geographical range. Results indicate that at least three sets of divergent Wolbachia strains invaded the genus Agelenopsis. After each invasion, the Wolbachia strains preferentially shuffled across species of this host genus by horizontal transfer rather than cospeciation. Decoupling of Wolbachia and host mitochondrial haplotype (mitotypes) evolutionary histories within single species reveals an extensive contribution of horizontal transfer also in the rapid dispersal of Wolbachia among conspecific host populations. These findings provide some of the strongest evidence to support the association of related Wolbachia strains with related hosts by means of both vertical and horizontal strain transmission. Similar analyses across a broader range of invertebrate taxa are needed, using sensitive methods for strain typing such as MLST, to determine if this pattern of Wolbachia dispersal is peculiar to Agelenopsis (or spiders), or is in fact a general pattern in arthropods.}, } @article {pmid18171476, year = {2008}, author = {Xi, Z and Gavotte, L and Xie, Y and Dobson, SL}, title = {Genome-wide analysis of the interaction between the endosymbiotic bacterium Wolbachia and its Drosophila host.}, journal = {BMC genomics}, volume = {9}, number = {}, pages = {1}, pmid = {18171476}, issn = {1471-2164}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cell Line ; Down-Regulation ; Drosophila/*genetics/immunology/*microbiology ; Female ; Gene Expression Profiling ; Genome, Insect/*genetics ; Gonads/enzymology ; Gram-Negative Bacterial Infections/genetics ; Heat-Shock Proteins/genetics ; Host-Pathogen Interactions/*genetics ; Male ; *Oligonucleotide Array Sequence Analysis ; Peptidyl-Dipeptidase A/genetics ; Phenotype ; Reproduction/genetics ; Sex Characteristics ; Symbiosis/*genetics ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Intracellular Wolbachia bacteria are obligate, maternally-inherited, endosymbionts found frequently in insects and other invertebrates. The success of Wolbachia can be attributed in part to an ability to alter host reproduction via mechanisms including cytoplasmic incompatibility (CI), parthenogenesis, feminization and male killing. Despite substantial scientific effort, the molecular mechanisms underlying the Wolbachia/host interaction are unknown.

RESULTS: Here, an in vitro Wolbachia infection was generated in the Drosophila S2 cell line, and transcription profiles of infected and uninfected cells were compared by microarray. Differentially-expressed patterns related to reproduction, immune response and heat stress response are observed, including multiple genes that have been previously reported to be involved in the Wolbachia/host interaction. Subsequent in vivo characterization of differentially-expressed products in gonads demonstrates that Angiotensin Converting Enzyme (Ance) varies between Wolbachia infected and uninfected flies and that the variation occurs in a sex-specific manner. Consistent with expectations for the conserved CI mechanism, the observed Ance expression pattern is repeatable in different Drosophila species and with different Wolbachia types. To examine Ance involvement in the CI phenotype, compatible and incompatible crosses of Ance mutant flies were conducted. Significant differences are observed in the egg hatch rate resulting from incompatible crosses, providing support for additional experiments examining for an interaction of Ance with the CI mechanism.

CONCLUSION: Wolbachia infection is shown to affect the expression of multiple host genes, including Ance. Evidence for potential Ance involvement in the CI mechanism is described, including the prior report of Ance in spermatid differentiation, Wolbachia-induced sex-specific effects on Ance expression and an Ance mutation effect on CI levels. The results support the use of Wolbachia infected cell cultures as an appropriate model for predicting in vivo host/Wolbachia interactions.}, } @article {pmid20302522, year = {2008}, author = {Sakamoto, H and Kageyama, D and Hoshizaki, S and Ishikawa, Y}, title = {Heat treatment of the Adzuki bean borer, Ostrinia scapulalis infected with wolbachia gives rise to sexually mosaic offspring.}, journal = {Journal of insect science (Online)}, volume = {8}, number = {}, pages = {1-5}, pmid = {20302522}, issn = {1536-2442}, mesh = {Animals ; Female ; *Hot Temperature ; Larva/microbiology ; Male ; *Mosaicism ; Moths/*microbiology ; Sex Ratio ; Survival Analysis ; Wolbachia/*physiology ; }, abstract = {A maternally inherited intracellular bacterium, Wolbachia, causes reproductive alterations in its arthropod hosts. In the adzuki bean borer, Ostrinia scapulalis (Walker) (Lepidoptera: Crambidae), naturally-occurring Wolbachia selectively kills male progeny. This Wolbachia strain appears to have a feminizing effect, since antibiotic treatment of infected female moths gives rise to male progeny with sexually mosaic phenotypes. It is proposed that male-specific death occurs through the feminizing effect, and sexual mosaics are produced when this effect is incompletely exerted. Here we examined whether the treatment of infected female moths with high temperatures (34 degrees C, 36 degrees C, or 38 degrees C), which is likely to suppress the activity of Wolbachia, induces sexually mosaic progeny. It was found that eggs laid within 24 h after treatment of Wolbachia-infected mothers at 36 degrees C gave rise to seven sexual mosaics along with 54 normal females. The time lag between treatment and the appearance of mosaic progeny was much shorter with heat treatment than antibiotic treatment, suggesting that heat treatment is more useful for spotting developmental timing when Wolbachia exerts its feminizing effect on O. scapulalis embryos.}, } @article {pmid19381483, year = {2008}, author = {Ahantarig, A and Trinachartvanit, W and Chauvatcharin, N and Kittayapong, P and Baimai, V}, title = {Wolbachia and bacteriophage WO-B density of Wolbachia A-infected Aedes albopictus mosquito.}, journal = {Folia microbiologica}, volume = {53}, number = {6}, pages = {547-550}, pmid = {19381483}, issn = {1874-9356}, mesh = {Aedes/growth & development/*microbiology/virology ; Animals ; Bacteriophages/*isolation & purification ; Female ; Larva/microbiology/virology ; Ovum/microbiology/virology ; Polymerase Chain Reaction ; Prophages/isolation & purification ; Pupa/microbiology/virology ; Reproduction ; Virus Replication ; Wolbachia/*isolation & purification/physiology/virology ; }, abstract = {Wolbachia are maternally inherited symbiotic bacteria capable of inducing an extensive range of reproductive abnormalities in their hosts, including cytoplasmic incompatibility (CI). Its density (concentration) is likely to influence the penetrance of CI in incompatible crosses. The variations of Wolbachia density could also be linked with phage WO density. We determined the relative density (relative concentration) of prophage WO orf7 and Wolbachia (phage-to-bacteria ratio) during early developmental and adult stages of singly infected Aedes albopictus mosquito (Wolbachia A-infected) by using real-time quantitative PCR. Phage WO and Wolbachia did not develop at the same rate. Relative Wolbachia density (bacteria-to-host ratio) was high later in development (adult stages) whilst relative prophage WO density (phage-to-bacteria ratio) was low in the adult stages. Furthermore, 12-d-old adults of singly infected female mosquito had the highest Wolbachia density. In contrast, the larval stage 4 (L4) contained the highest prophage WO-B orf7 density. The association of hosts-Wolbachia-phage among diverse species is different. Thus, if phage and Wolbachia are involved in CI mechanism, the information of this association should be acquired for each specific type of organism for future use of population replacement or gene drive system.}, } @article {pmid18155858, year = {2008}, author = {Cordaux, R}, title = {ISWpi1 from Wolbachia pipientis defines a novel group of insertion sequences within the IS5 family.}, journal = {Gene}, volume = {409}, number = {1-2}, pages = {20-27}, doi = {10.1016/j.gene.2007.10.035}, pmid = {18155858}, issn = {0378-1119}, mesh = {Amino Acid Sequence ; Base Sequence ; DNA Transposable Elements ; Evolution, Molecular ; Molecular Sequence Data ; Mutagenesis, Insertional ; Open Reading Frames ; Phylogeny ; Transposases/*genetics ; Wolbachia/*genetics ; }, abstract = {Insertion sequences are transposable elements that can represent substantial proportions of prokaryotic genomes and play a substantial role in shaping host genome evolution. As such, evaluating and understanding insertion sequence diversity is an important task to fulfill, because it is expected to yield new insight into the evolution of bacterial transposable elements and contribute to improve genome annotations. Here, I characterized an insertion sequence, termed ISWpi1, for which the taxonomic distribution appears to be restricted to the obligate intracellular alpha-Proteobacterium Wolbachia pipientis. ISWpi1 exhibits approximately 46% identity at the amino acid level with members of the IS1031 group of insertion sequences from the IS5 family. However, the IS1031 group is characterized by a transposase gene encoded by a single open reading frame, whereas the ISWpi1 transposase gene consists of two overlapping open reading frames presumably translated as a single protein via programmed translational frameshifting. Such structure suggests that ISWpi1 may instead be related to the IS427 group of insertion sequences from the IS5 family. Altogether, these data indicate that ISWpi1 extends the known spectrum of diversity of the IS5 family, and I propose to define a novel group of insertion sequences within the IS5 family typified by ISWpi1. Probable transpositional activity, relevant insertion site preferences and taxonomic specificity make ISWpi1 a promising tool for experimentally manipulating W. pipientis bacteria, especially in light of the increasing interest in developing these bacteria as tools for controlling insect disease vectors and agricultural pests.}, } @article {pmid18093008, year = {2007}, author = {Ballard, JW and Melvin, RG}, title = {Tetracycline treatment influences mitochondrial metabolism and mtDNA density two generations after treatment in Drosophila.}, journal = {Insect molecular biology}, volume = {16}, number = {6}, pages = {799-802}, doi = {10.1111/j.1365-2583.2007.00760.x}, pmid = {18093008}, issn = {0962-1075}, support = {R01 GM067862-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Adenosine Diphosphate/metabolism ; Animals ; Anti-Bacterial Agents/pharmacology ; Base Sequence ; Crosses, Genetic ; DNA Primers/genetics ; DNA, Mitochondrial/genetics/*metabolism ; Drosophila/*drug effects/genetics/*metabolism/microbiology ; Female ; Male ; Mitochondria/*drug effects/*metabolism ; Tetracycline/*pharmacology ; Wolbachia/pathogenicity ; }, abstract = {Tetracycline is commonly used to clear Wolbachia from infected insects. Studies then compare specific biochemical and/or life-history traits between infected and uninfected individuals with the same genetic background. We investigated the potential for tetracycline to influence mitochondrial efficiency and mitochondrial (mt)DNA density two generations after treatment in Drosophila simulans. We observed that antibiotic treatment resulted in a decline in inorganic phosphate incorporated into ATP per mole of oxygen consumed (ADP:O ratio). Further, tetracycline treatment caused a significant increase in mtDNA density in naturally Wolbachia-uninfected but not in naturally Wolbachia-infected lines suggesting a dosage effect. These data suggest that the current practice of comparing Wolbachia-infected and Wolbachia-uninfected insects two generations after tetracycline treatment needs to be re-evaluated.}, } @article {pmid18085821, year = {2007}, author = {Serbus, LR and Sullivan, W}, title = {A cellular basis for Wolbachia recruitment to the host germline.}, journal = {PLoS pathogens}, volume = {3}, number = {12}, pages = {e190}, pmid = {18085821}, issn = {1553-7374}, support = {F32 GM080192/GM/NIGMS NIH HHS/United States ; GM080192-01A1/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila Proteins/*metabolism ; Drosophila melanogaster/*microbiology ; Female ; *Host-Pathogen Interactions ; Infectious Disease Transmission, Vertical ; Insect Vectors/*microbiology ; Kinesins/*metabolism ; Microscopy, Confocal ; Microtubules/microbiology ; Oocytes/cytology/*microbiology/physiology ; Oogenesis ; Wolbachia/*pathogenicity/physiology ; }, abstract = {Wolbachia are among the most widespread intracellular bacteria, carried by thousands of metazoan species. The success of Wolbachia is due to efficient vertical transmission by the host maternal germline. Some Wolbachia strains concentrate at the posterior of host oocytes, which promotes Wolbachia incorporation into posterior germ cells during embryogenesis. The molecular basis for this localization strategy is unknown. Here we report that the wMel Wolbachia strain relies upon a two-step mechanism for its posterior localization in oogenesis. The microtubule motor protein kinesin-1 transports wMel toward the oocyte posterior, then pole plasm mediates wMel anchorage to the posterior cortex. Trans-infection tests demonstrate that factors intrinsic to Wolbachia are responsible for directing posterior Wolbachia localization in oogenesis. These findings indicate that Wolbachia can direct the cellular machinery of host oocytes to promote germline-based bacterial transmission. This study also suggests parallels between Wolbachia localization mechanisms and those used by other intracellular pathogens.}, } @article {pmid18076549, year = {2007}, author = {Debrah, AY and Mand, S and Marfo-Debrekyei, Y and Batsa, L and Pfarr, K and Buttner, M and Adjei, O and Buttner, D and Hoerauf, A}, title = {Macrofilaricidal effect of 4 weeks of treatment with doxycycline on Wuchereria bancrofti.}, journal = {Tropical medicine & international health : TM & IH}, volume = {12}, number = {12}, pages = {1433-1441}, doi = {10.1111/j.1365-3156.2007.01949.x}, pmid = {18076549}, issn = {1365-3156}, mesh = {Adolescent ; Adult ; Animals ; Antigens, Helminth/blood ; Antiparasitic Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Elephantiasis, Filarial/*drug therapy/immunology ; Ghana ; Humans ; Ivermectin/*therapeutic use ; Male ; Middle Aged ; Treatment Outcome ; Wuchereria bancrofti/*drug effects/isolation & purification/pathogenicity ; }, abstract = {OBJECTIVE: To evaluate the efficacy of doxycycline as a macrofilaricidal agent against Wuchereria bancrofti.

METHOD: In the Western Region of Ghana, 18 patients infected with W. bancrofti were recruited and treated with 200 mg doxycycline per day for 4 weeks. Seven untreated patients served as controls. Four months after doxycycline treatment, all patients received 150 mug/kg ivermectin. Patients were monitored for Wolbachia and microfilaria loads, antigenaemia and filarial dance sign (FDS).

RESULTS: Four months after doxycycline treatment, cases had a significantly lower Wolbachia load than controls; and 24 months after treatment, microfilaraemia, antigenaemia and frequency of FDS were significantly lower in cases than controls. Most importantly, 4 weeks of doxycycline killed 80% of macrofilariae, which is comparable with the results of a 6-week regimen. Circulating filarial antigenaemia and FDS were strongly correlated.

CONCLUSION: A 4-week regimen of doxycycline seems sufficient to kill adult W. bancrofti and could be advantageous for the treatment of individual patients, e.g. in outpatient clinics.}, } @article {pmid18073380, year = {2008}, author = {Nikoh, N and Tanaka, K and Shibata, F and Kondo, N and Hizume, M and Shimada, M and Fukatsu, T}, title = {Wolbachia genome integrated in an insect chromosome: evolution and fate of laterally transferred endosymbiont genes.}, journal = {Genome research}, volume = {18}, number = {2}, pages = {272-280}, pmid = {18073380}, issn = {1088-9051}, mesh = {Animals ; Base Sequence ; Blotting, Southern ; Chromosome Mapping ; Chromosomes/*genetics ; Coleoptera/genetics/*microbiology ; *Evolution, Molecular ; Gene Transfer, Horizontal/*genetics ; Genome/*genetics ; In Situ Hybridization, Fluorescence ; Likelihood Functions ; Models, Genetic ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Pseudogenes/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*genetics ; }, abstract = {Recent accumulation of microbial genome data has demonstrated that lateral gene transfers constitute an important and universal evolutionary process in prokaryotes, while those in multicellular eukaryotes are still regarded as unusual, except for endosymbiotic gene transfers from mitochondria and plastids. Here we thoroughly investigated the bacterial genes derived from a Wolbachia endosymbiont on the nuclear genome of the beetle Callosobruchus chinensis. Exhaustive PCR detection and Southern blot analysis suggested that approximately 30% of Wolbachia genes, in terms of the gene repertoire of wMel, are present on the insect nuclear genome. Fluorescent in situ hybridization located the transferred genes on the proximal region of the basal short arm of the X chromosome. Molecular evolutionary and other lines of evidence indicated that the transferred genes are probably derived from a single lateral transfer event. The transferred genes were, for the length examined, structurally disrupted, freed from functional constraints, and transcriptionally inactive. Hence, most, if not all, of the transferred genes have been pseudogenized. Notwithstanding this, the transferred genes were ubiquitously detected from Japanese and Taiwanese populations of C. chinensis, while the number of the transferred genes detected differed between the populations. The transferred genes were not detected from congenic beetle species, indicating that the transfer event occurred after speciation of C. chinensis, which was estimated to be one or several million years ago. These features of the laterally transferred endosymbiont genes are compared with the evolutionary patterns of mitochondrial and plastid genome fragments acquired by nuclear genomes through recent endosymbiotic gene transfers.}, } @article {pmid18069341, year = {2007}, author = {Aleksandrov, ID and Aleksandrova, MV and Goriacheva, II and Roshchina, NV and Shaĭkevich, EV and Zakharov, IA}, title = {[Removing endosymbiotic Wolbachia specifically decreases lifespan of females and competitiveness in a laboratory strain of Drosophila melanogaster].}, journal = {Genetika}, volume = {43}, number = {10}, pages = {1372-1378}, pmid = {18069341}, issn = {0016-6758}, mesh = {Aging ; Animals ; Drosophila melanogaster/*microbiology/*physiology ; Female ; *Longevity ; Male ; *Symbiosis ; Tetracycline/pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {To understand specific symbiotic relationships ensuring stable existing of the bacterium Wolbachia in laboratory strains of Drosophila melanogaster, the imago lifespan and senescence rate, as well as competitiveness, have been evaluated as components of fitness in females from the following laboratory strains: (1) inbred strain 95 infected with Wolbachia; (2) two uninfected strains obtained by tetracycline treatment that were genetically similar to strain 95; and (3) two control, uninfected, wild-type laboratory strains that were used to assess the possible effects of the antibiotic on the studied characters in the absence of Wolbachia. The results have shown that infected females have longer lifespan and competitiveness than females with the same genotype uninfected with Wolbachia. The increase in the senescence and mortality rates with age was also slower in infected females. It is noteworthy that tetracycline does not affect the lifespan of females from the two control, uninfected, wild-type strains. Therefore, the antibiotic is not the cause of the positive changes in fitness that were observed in infected females. The obtained results are the first direct evidence that the relationship s in the Wolbachia-D melanogaster symbiotic system are mutualistic rather than parasitic, at least in micropopulations adapted to laboratory conditions.}, } @article {pmid18067571, year = {2008}, author = {Champion de Crespigny, FE and Hurst, LD and Wedell, N}, title = {Do Wolbachia-associated incompatibilities promote polyandry?.}, journal = {Evolution; international journal of organic evolution}, volume = {62}, number = {1}, pages = {107-122}, doi = {10.1111/j.1558-5646.2007.00274.x}, pmid = {18067571}, issn = {0014-3820}, mesh = {Animals ; Biological Evolution ; Drosophila/*microbiology ; Female ; Male ; Reproduction ; Sexual Behavior, Animal/*physiology ; Wolbachia/*physiology ; }, abstract = {The genetic incompatibility avoidance hypothesis as an explanation for the polyandrous mating strategies (mating with more than one male) of females of many species has received significant attention in recent years. It has received support from both empirical studies and a meta-analysis, which concludes that polyandrous females enjoy increased reproductive success through improved offspring viability relative to monandrous females. In this study we investigate whether polyandrous female Drosophila simulans improve their fitness relative to monandrous females in the face of severe Wolbachia-associated reproductive incompatibilities. We use the results of this study to develop models that test the predictions that Wolbachia should promote polyandry, and that polyandry itself may constrain the spread of Wolbachia. Uniquely, our models allow biologically relevant rates of incompatibility to coevolve with a polyandry modifier allele, which allows us to evaluate the fate of the modifier and that of Wolbachia. Our empirical results reveal that polyandrous females significantly reduce the reproductive costs of Wolbachia, owing to infected males being poor sperm competitors. The models show that this disadvantage in sperm competition can inhibit or prevent the invasion of Wolbachia. However, despite the increased reproductive success obtained by polyandrous females, the spread of a polyandry modifier allele is constrained by any costs that might be associated with polyandry and the low frequency of incompatible matings when Wolbachia has reached a stable equilibrium. Therefore, although incompatibility avoidance may be a benefit of polyandry, our findings do not support the hypothesis that genetic incompatibilities caused by Wolbachia promote the evolution of polyandry.}, } @article {pmid18066693, year = {2008}, author = {Tiawsirisup, S and Sripatranusorn, S and Oraveerakul, K and Nuchprayoon, S}, title = {Distribution of mosquito (Diptera: Culicidae) species and Wolbachia (Rickettsiales: Rickettsiaceae) infections during the bird immigration season in Pathumthani province, central Thailand.}, journal = {Parasitology research}, volume = {102}, number = {4}, pages = {731-735}, pmid = {18066693}, issn = {0932-0113}, mesh = {*Animal Migration ; Animals ; Anopheles/microbiology ; Birds/*physiology ; Culex/microbiology ; *Culicidae/classification/microbiology/physiology ; Malvaceae/microbiology ; Seasons ; Species Specificity ; Thailand ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Mosquito distribution in the immigration bird-nested area, Pathumthani province, was investigated from August to December in 2006. Mosquitoes were collected by using CO2-baited Centers for Disease Control light traps in which dry ice was used as a source of CO2 to attract mosquitoes. Six traps were operated from 4 p.m. until 7 a.m. on each study day. Four genera, which were Anopheles, Armigeres, Culex, and Mansonia with 14 species of mosquitoes were collected. Culex gelidus (13.94-59.41%) and Culex tritaeniorhynchus (32.87-70.30%) were most collected species in this area for every month. Other two species with moderate distribution in this area were Anopheles barbirostris (0.76-3.30%) and Mansonia uniformis (1.55-11.36%). Polymerase Chain Reactions were performed for testing Wolbachia infection in Cx. gelidus and Cx. tritaeniorhynchus only. Fifty-four percent (15/28 pools) of Cx. gelidus and none (0/20 pools) of Cx. tritaeniorhynchus were positive for Wolbachia infection. Wolbachia infection in other mosquito species collected in this and other areas need to be investigated to understand species and geographic variation of Wolbachia infection in mosquitoes in nature.}, } @article {pmid18045498, year = {2007}, author = {Gupta, RS and Mok, A}, title = {Phylogenomics and signature proteins for the alpha proteobacteria and its main groups.}, journal = {BMC microbiology}, volume = {7}, number = {}, pages = {106}, pmid = {18045498}, issn = {1471-2180}, mesh = {Alphaproteobacteria/*chemistry/*classification/genetics ; Amino Acid Sequence ; Bacterial Proteins/*analysis ; Evolution, Molecular ; Molecular Sequence Data ; Open Reading Frames ; *Phylogeny ; Sequence Alignment ; }, abstract = {BACKGROUND: Alpha proteobacteria are one of the largest and most extensively studied groups within bacteria. However, for these bacteria as a whole and for all of its major subgroups (viz. Rhizobiales, Rhodobacterales, Rhodospirillales, Rickettsiales, Sphingomonadales and Caulobacterales), very few or no distinctive molecular or biochemical characteristics are known.

RESULTS: We have carried out comprehensive phylogenomic analyses by means of Blastp and PSI-Blast searches on the open reading frames in the genomes of several alpha-proteobacteria (viz. Bradyrhizobium japonicum, Brucella suis, Caulobacter crescentus, Gluconobacter oxydans, Mesorhizobium loti, Nitrobacter winogradskyi, Novosphingobium aromaticivorans, Rhodobacter sphaeroides 2.4.1, Silicibacter sp. TM1040, Rhodospirillum rubrum and Wolbachia (Drosophila) endosymbiont). These studies have identified several proteins that are distinctive characteristics of all alpha-proteobacteria, as well as numerous proteins that are unique repertoires of all of its main orders (viz. Rhizobiales, Rhodobacterales, Rhodospirillales, Rickettsiales, Sphingomonadales and Caulobacterales) and many families (viz. Rickettsiaceae, Anaplasmataceae, Rhodospirillaceae, Acetobacteraceae, Bradyrhiozobiaceae, Brucellaceae and Bartonellaceae). Many other proteins that are present at different phylogenetic depths in alpha-proteobacteria provide important information regarding their evolution. The evolutionary relationships among alpha-proteobacteria as deduced from these studies are in excellent agreement with their branching pattern in the phylogenetic trees and character compatibility cliques based on concatenated sequences for many conserved proteins. These studies provide evidence that the major groups within alpha-proteobacteria have diverged in the following order: (Rickettsiales(Rhodospirillales (Sphingomonadales (Rhodobacterales (Caulobacterales-Parvularculales (Rhizobiales)))))). We also describe two conserved inserts in DNA Gyrase B and RNA polymerase beta subunit that are distinctive characteristics of the Sphingomonadales and Rhodosprilllales species, respectively. The results presented here also provide support for the grouping of Hyphomonadaceae and Parvularcula species with the Caulobacterales and the placement of Stappia aggregata with the Rhizobiaceae group.

CONCLUSION: The alpha-proteobacteria-specific proteins and indels described here provide novel and powerful means for the taxonomic, biochemical and molecular biological studies on these bacteria. Their functional studies should prove helpful in identifying novel biochemical and physiological characteristics that are unique to these bacteria.}, } @article {pmid18043659, year = {2007}, author = {Pornwiroon, W and Kearney, MT and Husseneder, C and Foil, LD and Macaluso, KR}, title = {Comparative microbiota of Rickettsia felis-uninfected and -infected colonized cat fleas, Ctenocephalides felis.}, journal = {The ISME journal}, volume = {1}, number = {5}, pages = {394-402}, doi = {10.1038/ismej.2007.38}, pmid = {18043659}, issn = {1751-7362}, support = {AI60821/AI/NIAID NIH HHS/United States ; P20 RR0201595/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Cats ; Host-Pathogen Interactions ; Insect Vectors/*microbiology ; Rickettsia felis/*physiology ; Siphonaptera/*microbiology ; }, abstract = {Fleas serve as arthropod vectors for several emerging and re-emerging infectious disease causing agents including, Rickettsia felis. Although the prevalence of R. felis infection in colonies of fleas has been examined, the influence of the R. felis infection on flea microbiota has not been investigated. We identified three colonies of cat fleas, Ctenocephalides felis, with varying prevalence of R. felis infection (Louisiana State University (LSU), 93.8%; Professional Laboratory and Research Services Inc. (PLRS), 16.4%; Elward II (EL), 0%) and subsequently utilized polymerase chain reaction amplification, restriction fragment length polymorphism analysis and sequencing of the 1.4-kb portions of 16S rRNA genes to examine the diversity of bacteria in the flea populations. A total of 17 different bacterial 16S rRNA gene sequences were identified among the C. felis colonies. The prevalence of two Wolbachia species that were identified in each flea colony differed between colonies and R. felis-uninfected and -infected fleas. Species richness was unchanged among the R. felis-uninfected (LSU, PLRS and EL colonies) and -infected (LSU and PLRS colonies) fleas; however, between R. felis-uninfected and -infected fleas within both the LSU and PLRS colonies, R. felis-uninfected fleas have greater species richness. Diversity indices did not identify a difference in diversity between any of the flea samples. The interaction of endosymbionts within arthropods can widely impact the dissemination of vertically transmitted pathogenic bacteria; and the reciprocal may be true. These results suggest that carriage of R. felis has an impact on the richness of flea microbiota.}, } @article {pmid18005738, year = {2007}, author = {Blaxter, M}, title = {Symbiont genes in host genomes: fragments with a future?.}, journal = {Cell host & microbe}, volume = {2}, number = {4}, pages = {211-213}, doi = {10.1016/j.chom.2007.09.008}, pmid = {18005738}, issn = {1934-6069}, mesh = {Animals ; Cell Nucleus/genetics ; Gene Transfer, Horizontal ; Genes, Bacterial ; Nematoda/genetics/physiology ; Plant Roots/parasitology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {While lateral transfer is the rule in the evolutionary history of bacterial and archaeal genes, events of transfer from prokaryotes to eukaryotes are rare. Germline-transmitted animal symbionts, such as Wolbachia pipientis, are well placed to participate in such transfers. In a recent issue of Science, Dunning Hotopp et al. identified instances of transfer of Wolbachia DNA to host genomes. It is unknown whether these transfers represent innovation in animal evolution.}, } @article {pmid17999080, year = {2008}, author = {Hoerauf, A and Specht, S and Büttner, M and Pfarr, K and Mand, S and Fimmers, R and Marfo-Debrekyei, Y and Konadu, P and Debrah, AY and Bandi, C and Brattig, N and Albers, A and Larbi, J and Batsa, L and Taylor, MJ and Adjei, O and Büttner, DW}, title = {Wolbachia endobacteria depletion by doxycycline as antifilarial therapy has macrofilaricidal activity in onchocerciasis: a randomized placebo-controlled study.}, journal = {Medical microbiology and immunology}, volume = {197}, number = {3}, pages = {295-311}, pmid = {17999080}, issn = {0300-8584}, mesh = {Adult ; Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; Female ; Filaricides/*therapeutic use ; Ghana/epidemiology ; Humans ; Ivermectin/therapeutic use ; Male ; Middle Aged ; Onchocerca volvulus/*drug effects/microbiology ; Onchocerciasis/*drug therapy/parasitology ; Wolbachia/*drug effects ; }, abstract = {In a randomized, placebo-controlled trial in Ghana, 67 onchocerciasis patients received 200-mg/day doxycycline for 4-6 weeks, followed by ivermectin (IVM) after 6 months. After 6-27 months, efficacy was evaluated by onchocercoma histology, PCR and microfilariae determination. Administration of doxycycline resulted in endobacteria depletion and female worm sterilization. The 6-week treatment was macrofilaricidal, with >60% of the female worms found dead, despite the presence of new, Wolbachia-containing worms acquired after the administration of doxycycline. Doxycycline may be developed as second-line drug for onchocerciasis, to be administered in areas without transmission, in foci with IVM resistance and in areas with Loa co-infections.}, } @article {pmid17990528, year = {2007}, author = {Panteleev, DIu and Goriacheva, II and Andrianov, BV and Reznik, NL and Lazebnyĭ, OE and Kulikov, AM}, title = {[The endosymbiotic bacterium Wolbachia enhances the nonspecific resistance to insect pathogens and alters behavior of Drosophila melanogaster].}, journal = {Genetika}, volume = {43}, number = {9}, pages = {1277-1280}, pmid = {17990528}, issn = {0016-6758}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Beauveria/physiology ; Drosophila melanogaster/microbiology/*physiology ; Feeding Behavior ; Female ; Male ; Oviposition ; Sexual Behavior, Animal ; Symbiosis ; Tetracycline/*pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {To determine biologically important effects of the cytoplasmic endosymbiont Wolbachia, two substrains of the same Drosophila melanogaster strain have been studied, one of them infected with Wolbachia and the other treated with tetracycline to eliminate the bacterium. Female D. melanogaster infected with Wolbachia are more resistant to the fungus Bauveria bassiana (an insect pathogen) than uninfected females; infected females also exhibited changes in oviposition substrate preference. Males infected with the bacterium are more competitive than uninfected males. The possible role of Wolbachia in the formation of alternative ecological strategies of D. melanogaster is discussed.}, } @article {pmid17984346, year = {2007}, author = {Richards, FO and Amann, J and Arana, B and Punkosdy, G and Klein, R and Blanco, C and Lopez, B and Mendoza, C and Domínguez, A and Guarner, J and Maguire, JH and Eberhard, M}, title = {No depletion of Wolbachia from Onchocerca volvulus after a short course of rifampin and/or azithromycin.}, journal = {The American journal of tropical medicine and hygiene}, volume = {77}, number = {5}, pages = {878-882}, pmid = {17984346}, issn = {1476-1645}, mesh = {Adolescent ; Adult ; Animals ; Anthelmintics/therapeutic use ; Anti-Bacterial Agents/therapeutic use ; Azithromycin/*administration & dosage/*therapeutic use ; Child ; Child, Preschool ; Drug Therapy, Combination ; Female ; Humans ; Ivermectin/therapeutic use ; Male ; Middle Aged ; Onchocerca volvulus/drug effects/*microbiology ; Onchocerciasis/drug therapy/parasitology ; Rifampin/*administration & dosage/*therapeutic use ; Wolbachia/*drug effects ; }, abstract = {Endosymbionic Wolbachia bacteria inside adult Onchocerca volvulus worms (causing river blindness) are necessary for female worm fertility. We evaluated whether rifampin and/or azithromycin used in a five-day course could kill Wolbachia. In an open-label trial in Guatemala, 73 patients with 134 palpable onchocercal nodules were randomized into four treatment groups: rifampin, azithromycin, a combination of the two drugs, and controls (multivitamins). After five days of antibiotic treatment, all participants received a single dose of ivermectin on day 6. Nine months after treatment, the nodules were removed and the worms were examined. Skin snips to determine microfilariae were obtained at baseline and nine months. There were no significant differences between any of the treatment groups in the condition of the worms in the nodules, the presence of Wolbachia surface protein, or the number of microfilariae in skin. Short courses with these antibiotics will not clear Wolbachia from O. volvulus.}, } @article {pmid17983575, year = {2007}, author = {Bordenstein, SR}, title = {Evolutionary genomics: transdomain gene transfers.}, journal = {Current biology : CB}, volume = {17}, number = {21}, pages = {R935-6}, doi = {10.1016/j.cub.2007.09.022}, pmid = {17983575}, issn = {0960-9822}, mesh = {Animals ; Arthropods/*genetics/*microbiology ; Evolution, Molecular ; *Gene Transfer, Horizontal ; Genes, Bacterial ; Genomics ; Nematoda/*genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {Biologists have until now conceded that bacterial gene transfer to multicellular animals is relatively uncommon in Nature. A new study showing promiscuous insertions of bacterial endosymbiont genes into invertebrate genomes ushers in a shift in this paradigm.}, } @article {pmid17978845, year = {2007}, author = {Kluxen, G and Hörauf, A}, title = {[Ocular onchocerciasis: a key role for Wolbachia].}, journal = {Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft}, volume = {104}, number = {10}, pages = {860-865}, pmid = {17978845}, issn = {0941-293X}, mesh = {Animals ; Corneal Edema/immunology/microbiology ; Corneal Opacity/immunology/microbiology ; Disease Models, Animal ; Doxycycline/pharmacology ; Host-Parasite Interactions ; Humans ; Immunity, Innate/immunology ; Mice ; Microfilariae/immunology/*microbiology ; Neutrophils/immunology ; Onchocerca volvulus/immunology/*microbiology ; Onchocerciasis, Ocular/immunology/*microbiology ; Rickettsiaceae Infections/immunology/*microbiology ; *Symbiosis ; Virulence/drug effects/immunology ; Wolbachia/drug effects/immunology/*pathogenicity ; }, abstract = {Onchocerciasis is caused by the parasitic worm Onchocerca volvulus, which releases millions of offspring (microfilariae). Microfilariae migrate through the skin and can enter the anterior or posterior regions of the eye. While alive, the microfilariae appear to cause little or no inflammation, being in the anterior chamber. However, when they die, either by natural attrition or after chemotherapy, the host response to degenerating worms can result in ocular inflammation (keratitis, uveitis, chorioretinitis, neuritis of the optic nerve) that causes progressive loss of vision and ultimately leads to blindness. With the use of a mouse model of corneal inflammation to study the pathogenesis of ocular onchocerciasis by injecting worm extracts directly into the corneal stroma, it was found that worms treated with the antibiotic doxycycline, which destroys Wolbachia, induced lower corneal stromal thickness and stromal haze (indicators of corneal oedema and opacity) and neutrophil infiltration compared with both untreated worms and worms that do not harbour Wolbachia. These data indicate that endosymbiotic Wolbachia bacteria in filarial parasites have a key role in the pathogenesis of river blindness. Worms recovered from patients treated for 6 weeks with doxycycline contained fewer Wolbachia bacteria and had abnormal embryogenesis, indicating a role for Wolbachia in the survival or fecundity of the worms. Antibiotic treatment may also reduce the severity of the inflammatory response in the cornea.}, } @article {pmid17978237, year = {2007}, author = {Lau, KWK and Ren, J and Fung, MC and Woo, PCY and Yuen, KY and Chan, KKM and Qian, PY and Wong, PK and Wu, M}, title = {Fangia hongkongensis gen. nov., sp. nov., a novel gammaproteobacterium of the order Thiotrichales isolated from coastal seawater of Hong Kong.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {57}, number = {Pt 11}, pages = {2665-2669}, doi = {10.1099/ijs.0.65156-0}, pmid = {17978237}, issn = {1466-5026}, mesh = {Bacterial Typing Techniques ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Fatty Acids/analysis ; Gammaproteobacteria/*classification/genetics/*isolation & purification/physiology ; Genes, rRNA ; Hong Kong ; Molecular Sequence Data ; Phenotype ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Seawater/*microbiology ; Sequence Analysis, DNA ; Species Specificity ; }, abstract = {A Gram-negative, coccobacillus-shaped, aerobic bacterium, designated strain UST040201-002T, was isolated in February 2004 from seawater at the outlet of a sandfilter in Port Shelter, Hong Kong SAR, China. This strain possessed ubiquinone-8; its 16S rRNA gene sequence shared only 91% similarity with the sequence from Caedibacter taeniospiralis and 89-90% similarity with sequences from Francisella tularensis, Francisella novicida, Francisella philomiragia and Wolbachia persica. 16S rRNA gene sequence analysis showed that the strain formed a distinct clade with C. taeniospiralis. This subcluster formed a tight coherent group with members of the family Francisellaceae and W. persica. Combined phylogenetic and physiological data suggest that strain UST040201-002T represents a novel genus and species within the order Tauhiotrichales. The name Fangia hongkongensis gen. nov., sp. nov. is proposed; the type strain is UST040201-002T (=JCM 14605T=NRRL B-41860T).}, } @article {pmid17925215, year = {2007}, author = {Martin, OY and Gage, MJ}, title = {Evolutionary conflicts: rapid suppression of a male-killer.}, journal = {Current biology : CB}, volume = {17}, number = {19}, pages = {R849-51}, doi = {10.1016/j.cub.2007.07.049}, pmid = {17925215}, issn = {0960-9822}, mesh = {Animals ; *Biological Evolution ; Butterflies/*microbiology ; Female ; Male ; *Sex Determination Processes ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Conflicts between and within species can drive fast evolutionary change. A recent study has documented remarkably rapid counter-adaptations in the wild to an extreme sex-ratio distortion caused by a bacterial symbiont.}, } @article {pmid17919582, year = {2008}, author = {Lamb, TJ and Harris, A and Le Goff, L and Read, AF and Allen, JE}, title = {Litomosoides sigmodontis: vaccine-induced immune responses against Wolbachia surface protein can enhance the survival of filarial nematodes during primary infection.}, journal = {Experimental parasitology}, volume = {118}, number = {2}, pages = {285-289}, doi = {10.1016/j.exppara.2007.08.011}, pmid = {17919582}, issn = {0014-4894}, support = {G9901118/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Analysis of Variance ; Animals ; Bacterial Outer Membrane Proteins/*immunology ; Bacterial Vaccines/*immunology ; Cytokines/metabolism ; Female ; Filariasis/immunology/*parasitology ; Filarioidea/microbiology/*physiology ; Immunoglobulin G/blood ; Male ; Mice ; Mice, Inbred BALB C ; Symbiosis ; Vaccination ; Wolbachia/*immunology ; }, abstract = {Wolbachia are bacteria present within the tissues of most filarial nematodes. Filarial nematode survival is known to be affected by immune responses generated during filarial nematode infection and immune responses to Wolbachia can be found in different species harbouring filarial nematode infections, including humans. Using the rodent filarial model Litomosoides sigmodontis, we show that pre-exposure to wolbachia surface protein in a Th1 context (but not in a Th2-context) enhances worm survival on subsequent challenge. This study suggests that despite abundant evidence that pro-inflammatory reactions to the endosymbiont have detrimental effects on the both the nematode and mammalian host, they may under some circumstances be beneficial to the nematode.}, } @article {pmid17915272, year = {2008}, author = {Engelstädter, J and Telschow, A and Yamamura, N}, title = {Coexistence of cytoplasmic incompatibility and male-killing-inducing endosymbionts, and their impact on host gene flow.}, journal = {Theoretical population biology}, volume = {73}, number = {1}, pages = {125-133}, doi = {10.1016/j.tpb.2007.08.004}, pmid = {17915272}, issn = {0040-5809}, mesh = {Animal Migration ; Animals ; Butterflies/*parasitology ; Cytoplasm/*genetics ; Gene Flow/*genetics ; Male ; Models, Statistical ; Sex Ratio ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Male-killing (MK) and cytoplasmic incompatibility (CI) inducing bacteria are among the most common endosymbionts of arthropods. Previous theoretical research has demonstrated that these two types of endosymbionts cannot stably coexist within a single unstructured host population if no doubly infected host individuals occur. Here, we analyse a model of two host subpopulations connected by migration. We demonstrate that coexistence of MK- and CI-inducing endosymbionts is possible if migration rates are sufficiently low. In particular, our results suggest that for coexistence to be possible, migration rates into the subpopulation infected predominantly with MK-inducing endosymbionts must be considerably low, while migration rates from the MK- to the CI-infected subpopulation can be very high. We also analyse how the presence of MK- and CI-inducing endosymbionts affects host gene flow between the two subpopulations. Employing the concept of the 'effective migration rate', we demonstrate that compared with an uninfected subdivided population, gene flow is increased towards the MK-infected island, but decreased towards the CI-infected island. We discuss our results with respect to the butterfly Hypolimnas bolina, in which infection polymorphism of CI- and MK-inducing Wolbachia has been reported across South-Pacific island populations.}, } @article {pmid17899809, year = {2007}, author = {Ilinskiĭ, IuIu and Zakharov, IK}, title = {[The endosymbiont Wolbachia in Eurasian populations of Drosophila melanogaster].}, journal = {Genetika}, volume = {43}, number = {7}, pages = {905-915}, pmid = {17899809}, issn = {0016-6758}, mesh = {Animals ; Asia, Central ; Drosophila melanogaster/*microbiology ; Europe, Eastern ; Genetics, Population ; Genotype ; Transcaucasia ; Wolbachia/*genetics/isolation & purification/physiology ; }, abstract = {The endosymbiotic [alpha]-proteobacteria Wolbachia is widely spread among arthropods and Filariidae nematodes. This bacterium is transmitted vertically via a transovarian route. Wolbachia is a cause of several reproductive abnormalities in the host species. We analyzed the isofemale lines created using flies collected from Drosophila melanogaster natural populations for infection with the endosymbiont Wolbachia. Wolbachia were genotyped according to five variable markers: the presence of insertion sequence IS5 in two loci, the copy number of two minisatellite repeats, and an inversion. Overall, 665 isofemale lines isolated from the populations of D. melanogaster from Ukraine, Belarus, Moldova, Caucasus, Central Asia, Ural, Udmurtia, Altai, West and East Siberia, and Far East in 1974 through 2005 were used in the work. The samples from Ukrainian, Altaian, and Middle Asian populations were largest. The infection rate of D. melanogaster populations from Middle Asia, Altaian, and Eastern Europe (Ukraine, Moldavia, and Belarus) with Wolbachia amounted to 64, 56, and 39%, respectively. The D. melanogaster population from the Caucasus displayed heterogeneity in the genotypes of this cytoplasmic infection. The Wolbachia genotype wMel, detected in all the populations studied, was the most abundant. The genotype wMelCS2 was always present in the populations from Middle Asia and Altai and was among the rare variants in the D. melanogaster populations from the Eastern Europe. Single instances of the Wolbachia genotype wMelCS occurred in a few flies from the Central Asian and Altai populations, but was not found this genotype in the other regions.}, } @article {pmid17893742, year = {2007}, author = {Sakamoto, H and Kageyama, D and Hoshizaki, S and Ishikawa, Y}, title = {Sex-specific death in the Asian corn borer moth (Ostrinia furnacalis) infected with Wolbachia occurs across larval development.}, journal = {Genome}, volume = {50}, number = {7}, pages = {645-652}, doi = {10.1139/g07-041}, pmid = {17893742}, issn = {0831-2796}, mesh = {Animals ; Female ; Genotype ; Larva/growth & development ; Male ; Moths/*growth & development/*microbiology ; Sex Chromosomes ; Sex Factors ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Maternally inherited endosymbiotic bacteria of the genus Wolbachia induce various kinds of reproductive alterations in their arthropod hosts. In a Wolbachia-infected strain of the adzuki bean borer moth, Ostrinia scapulalis (Lepidoptera: Crambidae), males selectively die during larval development, while females selectively die when Wolbachia are eliminated by antibiotic treatment. We found that naturally occurring Wolbachia in the congener O. furnacalis caused sex-specific lethality similar to that in O. scapulalis. Cytogenetic analyses throughout the entire larval development clarified that the death of males (when infected) and females (when cured) took place mainly during early larval stages. However, some individuals also died after complete formation of larval bodies but before egg hatching, or at late larval stages, even in the penultimate instar. Although the specific timing was highly variable, death of males and females occurred before pupation without exception. The potential association of sex-specific lethality with the sex determination mechanism was also examined and is discussed.}, } @article {pmid17887971, year = {2007}, author = {Hayashi, TI and Marshall, JL and Gavrilets, S}, title = {The dynamics of sexual conflict over mating rate with endosymbiont infection that affects reproductive phenotypes.}, journal = {Journal of evolutionary biology}, volume = {20}, number = {6}, pages = {2154-2164}, doi = {10.1111/j.1420-9101.2007.01429.x}, pmid = {17887971}, issn = {1010-061X}, support = {GM56693/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Female ; Male ; Orthoptera/genetics/*microbiology/*physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Maternally inherited endosymbionts have been implicated as significant drivers of sexual conflict within their hosts, typically through sex-ratio manipulation. Empirical studies show that some of these endosymbionts have the potential to influence sexual conflict not by sex-ratio distortion, but by altering reproductive traits within their hosts. Research has already shown that reproductive traits involved in mating/fertilization process are integral 'players' in sexual conflict, thus suggesting the novel hypothesis that endosymbiont-induced changes in reproductive phenotypes can impact the dynamics of sexual conflict. Here, we use a standard quantitative genetic approach to model the effects of endosymbiont-induced changes in a female reproductive trait on the dynamics of sexual conflict over mating/fertilization rate. Our model shows that an endosymbiont-induced alteration of a host female reproductive trait that affects mating rate can maintain the endosymbiont infection within the host population, and does so in the absence of sex-ratio distortion and cytoplasmic incompatibility.}, } @article {pmid17885136, year = {2007}, author = {Ghedin, E and Wang, S and Spiro, D and Caler, E and Zhao, Q and Crabtree, J and Allen, JE and Delcher, AL and Guiliano, DB and Miranda-Saavedra, D and Angiuoli, SV and Creasy, T and Amedeo, P and Haas, B and El-Sayed, NM and Wortman, JR and Feldblyum, T and Tallon, L and Schatz, M and Shumway, M and Koo, H and Salzberg, SL and Schobel, S and Pertea, M and Pop, M and White, O and Barton, GJ and Carlow, CK and Crawford, MJ and Daub, J and Dimmic, MW and Estes, CF and Foster, JM and Ganatra, M and Gregory, WF and Johnson, NM and Jin, J and Komuniecki, R and Korf, I and Kumar, S and Laney, S and Li, BW and Li, W and Lindblom, TH and Lustigman, S and Ma, D and Maina, CV and Martin, DM and McCarter, JP and McReynolds, L and Mitreva, M and Nutman, TB and Parkinson, J and Peregrín-Alvarez, JM and Poole, C and Ren, Q and Saunders, L and Sluder, AE and Smith, K and Stanke, M and Unnasch, TR and Ware, J and Wei, AD and Weil, G and Williams, DJ and Zhang, Y and Williams, SA and Fraser-Liggett, C and Slatko, B and Blaxter, ML and Scott, AL}, title = {Draft genome of the filarial nematode parasite Brugia malayi.}, journal = {Science (New York, N.Y.)}, volume = {317}, number = {5845}, pages = {1756-1760}, pmid = {17885136}, issn = {1095-9203}, support = {R15 ES013128/ES/NIEHS NIH HHS/United States ; R01 LM006845-08/LM/NLM NIH HHS/United States ; R01 LM007938/LM/NLM NIH HHS/United States ; R01 AI048562-09/AI/NIAID NIH HHS/United States ; R01 LM006845/LM/NLM NIH HHS/United States ; R15 ES013128-01/ES/NIEHS NIH HHS/United States ; U01-AI50903/AI/NIAID NIH HHS/United States ; R01 LM007938-04/LM/NLM NIH HHS/United States ; U01 AI048828/AI/NIAID NIH HHS/United States ; R01 AI048562/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/*genetics/physiology ; Caenorhabditis/genetics ; Drosophila melanogaster/genetics ; Drug Resistance/genetics ; Filariasis/parasitology ; *Genome, Helminth ; Humans ; Molecular Sequence Data ; }, abstract = {Parasitic nematodes that cause elephantiasis and river blindness threaten hundreds of millions of people in the developing world. We have sequenced the approximately 90 megabase (Mb) genome of the human filarial parasite Brugia malayi and predict approximately 11,500 protein coding genes in 71 Mb of robustly assembled sequence. Comparative analysis with the free-living, model nematode Caenorhabditis elegans revealed that, despite these genes having maintained little conservation of local synteny during approximately 350 million years of evolution, they largely remain in linkage on chromosomal units. More than 100 conserved operons were identified. Analysis of the predicted proteome provides evidence for adaptations of B. malayi to niches in its human and vector hosts and insights into the molecular basis of a mutualistic relationship with its Wolbachia endosymbiont. These findings offer a foundation for rational drug design.}, } @article {pmid17883830, year = {2007}, author = {Walker, T and Klasson, L and Sebaihia, M and Sanders, MJ and Thomson, NR and Parkhill, J and Sinkins, SP}, title = {Ankyrin repeat domain-encoding genes in the wPip strain of Wolbachia from the Culex pipiens group.}, journal = {BMC biology}, volume = {5}, number = {}, pages = {39}, pmid = {17883830}, issn = {1741-7007}, support = {/WT_/Wellcome Trust/United Kingdom ; 079059/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Ankyrin Repeat ; Culex/*microbiology/physiology ; DNA Primers ; *Gene Expression Regulation, Bacterial ; *Genes, Bacterial ; Host-Pathogen Interactions ; Reverse Transcriptase Polymerase Chain Reaction ; Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {BACKGROUND: Wolbachia are obligate endosymbiotic bacteria maternally transmitted through the egg cytoplasm that are responsible for several reproductive disorders in their insect hosts, such as cytoplasmic incompatibility (CI) in infected mosquitoes. Species in the Culex pipiens complex display an unusually high number of Wolbachia-induced crossing types, and based on present data, only the wPip strain is present.

RESULTS: The sequencing of the wPip strain of Wolbachia revealed the presence of 60 ankyrin repeat domain (ANK) encoding genes and expression studies of these genes were carried out in adult mosquitoes. One of these ANK genes, pk2, is shown to be part of an operon of three prophage-associated genes with sex-specific expression, and is present in two identical copies in the genome. Another homolog of pk2 is also present that is differentially expressed in different Cx. pipiens group strains. A further two ANK genes showed sex-specific regulation in wPip-infected Cx. pipiens group adults.

CONCLUSION: The high number, variability and differential expression of ANK genes in wPip suggest an important role in Wolbachia biology, and the gene family provides both markers and promising candidates for the study of reproductive manipulation.}, } @article {pmid17879233, year = {2007}, author = {Pankewitz, F and Zöllmer, A and Gräser, Y and Hilker, M}, title = {Anthraquinones as defensive compounds in eggs of Galerucini leaf beetles: biosynthesis by the beetles?.}, journal = {Archives of insect biochemistry and physiology}, volume = {66}, number = {2}, pages = {98-108}, doi = {10.1002/arch.20215}, pmid = {17879233}, issn = {0739-4462}, mesh = {Animals ; Anthraquinones/*metabolism ; Coleoptera/*metabolism/microbiology ; DNA, Bacterial/*analysis ; DNA, Fungal/*analysis ; Female ; Ovum/chemistry/*metabolism/microbiology ; }, abstract = {Eggs of leaf beetles of the tribe Galerucini, subfamily Galerucinae, contain polyketides that are unusual in insects: 1,8-dihydroxylated anthraquinones (chrysazin, chrysophanol) and anthrones (dithranol, chrysarobin) deterring predators. The host plants do not contain these compounds. In the present study, we tested the hypothesis that the beetles, but not bacterial or fungal microorganisms living as endosymbionts within the beetles, produce the anthraquinones. The tansy leaf beetle Galeruca tanaceti was used as Galerucini model organism. It was treated with antimicrobial substances to eradicate the microorganisms and inhibit the hypothesised endosymbiotic anthraquinone production. Despite treatment, female G. tanaceti laid eggs containing anthraquinones. Although broad spectrum antimicrobials were used, it cannot be excluded that the potential endosymbiotic microorganisms are resistant. Given that the hypothesised endosymbionts are transferred via the eggs from one generation to the next, bacterial or fungal DNA was expected to be present in the eggs. With the exception of Wolbachia pipientis, however, no further 16S rDNA from bacteria responsible for anthraquinone biosynthesis was detected in eggs of untreated beetles. Because Wolbachia were also found in closely related anthraquinone-free insects, we exclude these bacteria as producers of the defensive polyketides. Nor was any 18S rDNA from fungi with anthraquinone biosynthetic abilities detected. Our results indicate that anthraquinones and anthrones in eggs of Galerucini are produced by beetle enzymes and not by endosymbiotic microorganisms within the eggs.}, } @article {pmid17878145, year = {2007}, author = {Weinert, LA and Tinsley, MC and Temperley, M and Jiggins, FM}, title = {Are we underestimating the diversity and incidence of insect bacterial symbionts? A case study in ladybird beetles.}, journal = {Biology letters}, volume = {3}, number = {6}, pages = {678-681}, pmid = {17878145}, issn = {1744-9561}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Bacteria/classification/*isolation & purification ; *Biodiversity ; Coleoptera/*microbiology ; Female ; Flavobacterium/classification/isolation & purification ; Male ; Rickettsia/classification/isolation & purification ; Sex Factors ; Sex Ratio ; Spiroplasma/classification/isolation & purification ; *Symbiosis ; Wolbachia/classification/isolation & purification ; }, abstract = {Vertically transmitted bacterial symbionts are common in arthropods. However, estimates of their incidence and diversity are based on studies that test for a single bacterial genus and often only include small samples of each host species. Focussing on ladybird beetles, we collected large samples from 21 species and tested them for four different bacterial symbionts. Over half the species were infected, and there were often multiple symbionts in the same population. In most cases, more females than males were infected, suggesting that the symbionts may be sex ratio distorters. Many of these infections would have been missed in previous studies as they only infect a small proportion of the population. Furthermore, 11 out of the 17 symbionts discovered by us were either in the genus Rickettsia or Spiroplasma, which are rarely sampled. Our results suggest that the true incidence and diversity of bacterial symbionts in insects may be far greater than previously thought.}, } @article {pmid17875630, year = {2007}, author = {Gillette-Ferguson, I and Daehnel, K and Hise, AG and Sun, Y and Carlson, E and Diaconu, E and McGarry, HF and Taylor, MJ and Pearlman, E}, title = {Toll-like receptor 2 regulates CXC chemokine production and neutrophil recruitment to the cornea in Onchocerca volvulus/Wolbachia-induced keratitis.}, journal = {Infection and immunity}, volume = {75}, number = {12}, pages = {5908-5915}, pmid = {17875630}, issn = {1098-5522}, support = {P30 EY011373/EY/NEI NIH HHS/United States ; K08 AI054652/AI/NIAID NIH HHS/United States ; R01 EY010320/EY/NEI NIH HHS/United States ; T32 AI007024/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; EY10320/EY/NEI NIH HHS/United States ; AI-07024/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bone Marrow Cells/immunology ; Cell Line ; Chemokines, CXC/biosynthesis/*immunology ; Cornea/immunology/pathology ; Female ; Keratitis/*immunology/parasitology ; Male ; Mice ; Mice, Inbred C57BL ; Neutrophils/*immunology ; Onchocerca volvulus/*immunology ; Onchocerciasis, Ocular/*immunology/parasitology ; Toll-Like Receptor 2/*immunology ; Wolbachia/*immunology ; }, abstract = {The filarial nematode Onchocerca volvulus is the causative organism of river blindness. Our previous studies demonstrated an essential role for endosymbiotic Wolbachia bacteria in corneal disease, which is characterized by neutrophil infiltration into the corneal stroma and the development of corneal haze. To determine the role of Toll-like receptors (TLRs) in neutrophil recruitment and activation, we injected a soluble extract of O. volvulus containing Wolbachia bacteria into the corneal stromata of C57BL/6, TLR2-/-, TLR4-/-, TLR2/4-/-, and TLR9-/- mice. We found an essential role for TLR2, but not TLR4 or TLR9, in neutrophil recruitment to the cornea and development of corneal haze. Furthermore, chimeric mouse bone marrow studies showed that resident bone marrow-derived cells in the cornea can initiate this response. TLR2 expression was also essential for CXC chemokine production by resident cells in the cornea, including corneal fibroblasts, and for neutrophil activation. Taken together, these findings indicate that Wolbachia activates TLR2 on resident bone marrow-derived cells in the corneal stroma to produce CXC chemokines, leading to neutrophil recruitment to the corneal stroma, and that TLR2 mediates O. volvulus/Wolbachia-induced neutrophil activation and development of corneal haze.}, } @article {pmid17849905, year = {2007}, author = {Dong, P and Wang, JJ and Hu, F and Jia, FX}, title = {Influence of wolbachia infection on the fitness of the stored-product pest Liposcelis tricolor (Psocoptera: Liposcelididae).}, journal = {Journal of economic entomology}, volume = {100}, number = {4}, pages = {1476-1481}, doi = {10.1603/0022-0493(2007)100[1476:iowiot]2.0.co;2}, pmid = {17849905}, issn = {0022-0493}, mesh = {Animals ; Fertility ; Insecta/growth & development/*microbiology/physiology ; Longevity ; Nymph/growth & development/microbiology/physiology ; Ovum/growth & development/microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacteria (Rickettsiaceae) that infect a wide range of arthropods and nematodes and that are associated with various reproductive abnormalities in their hosts. In this study, the effects of removal of Wolbachia infection on development, survival, and reproduction of Liposcelis tricolor Badonnel (Psocoptera: Liposcelididae) were investigated in laboratory. The Wolbachia-free strain was obtained by the removal of Wolbachia infection by using 1% rifampicin treatment on the Wolbachia-infected strain (control) for 4 wk, and no Wolbachia gene product was detected in this strain throughout the experiment. The results showed that the removal of Wolbachia infection had negative effects on the fitness of L. tricolor. Compared with the control strain, the Wolbachia-free strain (both in the first [F1] and second [F2] generation) had prolonged developmental times, reduced survivorship of immature stages, and reduced fecundity and longevity, resulting in much smaller rm values. Using rm values, the fitness for Wolbachia-free F1 and F2 relative to the control were calculated as 0.45 and 0.27, respectively. The results of this study further confirmed our previous conclusion that Wolbachia infection have positive effects on fecundity and fertility of L. tricolor, and for optimal reproduction of L. tricolor, Wolbachia must be present in psocids.}, } @article {pmid17825952, year = {2007}, author = {Aanen, DK and Hoekstra, RF}, title = {The evolution of obligate mutualism: if you can't beat 'em, join 'em.}, journal = {Trends in ecology & evolution}, volume = {22}, number = {10}, pages = {506-509}, doi = {10.1016/j.tree.2007.08.007}, pmid = {17825952}, issn = {0169-5347}, mesh = {Apoptosis ; *Biological Evolution ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is best known as a facultative endosymbiotic parasite, manipulating host reproduction. However, it has also evolved as an obligate mutualist at least twice. In a recent paper, Pannebakker et al. identify a possible mechanism for such a transition from facultative parasitism to obligate mutualism in a parasitic wasp in which Wolbachia are required for producing eggs (oogenesis). Their proposed mechanism suggests that compensatory evolution in the host to counter the harmful effects of Wolbachia is the basis of this evolutionary transition.}, } @article {pmid17822801, year = {2007}, author = {Darby, AC and Cho, NH and Fuxelius, HH and Westberg, J and Andersson, SG}, title = {Intracellular pathogens go extreme: genome evolution in the Rickettsiales.}, journal = {Trends in genetics : TIG}, volume = {23}, number = {10}, pages = {511-520}, doi = {10.1016/j.tig.2007.08.002}, pmid = {17822801}, issn = {0168-9525}, mesh = {Alphaproteobacteria/*genetics/*pathogenicity ; Bacterial Proteins ; *Evolution, Molecular ; Genes, Bacterial ; *Genome, Bacterial ; Humans ; Models, Genetic ; Orientia tsutsugamushi/genetics ; Rickettsia/genetics ; Virulence ; Wolbachia/genetics ; }, abstract = {The Rickettsiales, a genetically diverse group of the alpha-Proteobacteria, include major mammalian pathogens, such as the agents of epidemic typhus, scrub typhus, ehrlichioses and heartwater disease. Sequenced genomes of this bacterial order have provided exciting insights into reductive genome evolution, antigenic variation and host cell manipulation. Recent results suggest that human pathogens emerged relatively late in the evolution of the Rickettsiales. Surprisingly, there is no association between pathogenicity and the acquisition of novel virulence genes. Here, we explore the genomic differences between members of the Rickettsiales and ask what are the changes that enable infectious agents to emerge from seemingly harmless bacteria.}, } @article {pmid17805256, year = {2007}, author = {Callaway, E}, title = {Genomes within genomes.}, journal = {Nature}, volume = {449}, number = {7158}, pages = {6}, doi = {10.1038/449006b}, pmid = {17805256}, issn = {1476-4687}, mesh = {Animals ; Drosophila/classification/*genetics/*microbiology ; Female ; Gene Expression ; Gene Flow ; Genes, Bacterial/genetics ; Genome, Bacterial/*genetics ; Genome, Insect/*genetics ; Genomics ; Wolbachia/*genetics ; }, } @article {pmid17785193, year = {2007}, author = {Huang, Y and Magori, K and Lloyd, AL and Gould, F}, title = {Introducing transgenes into insect populations using combined gene-drive strategies: modeling and analysis.}, journal = {Insect biochemistry and molecular biology}, volume = {37}, number = {10}, pages = {1054-1063}, pmid = {17785193}, issn = {0965-1748}, support = {R01 AI054954/AI/NIAID NIH HHS/United States ; R01 AI054954-01A2/AI/NIAID NIH HHS/United States ; R01-AI54954-0IA2/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Animals, Genetically Modified ; Computer Simulation ; Female ; Insect Control ; Insect Vectors/*genetics ; Male ; Meiosis/genetics ; *Models, Genetic ; *Transgenes ; Wolbachia/genetics ; }, abstract = {Engineered underdominance (EU), meiotic drive (MD) and Wolbachia have been proposed as mechanisms for driving anti-pathogen transgenes into natural populations of insect vectors of human diseases. EU can drive transgenes to high and stable frequencies but requires the release of sizeable numbers of engineered insects. MD and Wolbachia either cannot maintain high frequencies of transgenes or lack appropriate expression in critical tissues, but both can drive the transgenes to spread from very low initial frequencies. Here we use mathematical models to assess the utility of combining EU with MD or with Wolbachia. Under some conditions, the combination of EU and MD results in a more efficient transgene-drive strategy than either mechanism alone. This combined strategy could drive the transgenes to stable fixation and would require fewer released insects than EU alone, especially when only males are released. However, a combination of EU and Wolbachia does not work better than EU alone because it requires the release of even more engineered insects.}, } @article {pmid17767593, year = {2007}, author = {Jaenike, J}, title = {Spontaneous emergence of a new Wolbachia phenotype.}, journal = {Evolution; international journal of organic evolution}, volume = {61}, number = {9}, pages = {2244-2252}, doi = {10.1111/j.1558-5646.2007.00180.x}, pmid = {17767593}, issn = {0014-3820}, mesh = {Animals ; Breeding ; DNA, Mitochondrial ; Drosophila/genetics/growth & development/*microbiology ; Female ; Genes, Insect ; Genotype ; Male ; *Phenotype ; Sex Chromosomes ; *Sex Ratio ; Species Specificity ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are among the most widespread symbionts on the earth. They spread within populations by various means of manipulating host reproduction, including cytoplasmic incompatibility (CI), male-killing (MK), parthenogenesis, and feminization. Phylogenetic analyses indicate that Wolbachia have the potential to undergo rapid evolutionary change in phenotype, for example, from CI to MK, although such analyses do not reveal the rate at which such transitions occur, nor the nature of the intermediate phenotypes. Here I show that a transition from CI to MK can occur almost instantaneously on an evolutionary time scale. A Wolbachia strain that causes CI in its natural host, Drosophila recens, was introgressed to its sister species D. subquinaria via the natural processes of hybridization and backcrossing. In some strains of D. subquinaria, infection with this Wolbachia strain caused essentially complete MK, resulting in all-female broods, whereas in other strains, there was no effect on offspring sex ratio. Crosses within and between D. subquinaria and D. recens revealed that resistance to MK is dominant, autosomal, multigenic, and dependent on zygotic, not maternal, genotype. MK in D. subquinaria is unusual in that the male offspring of infected females die during the larval stage, not as embryos. These findings suggest that MK and CI may share a similar underlying molecular basis.}, } @article {pmid17762140, year = {2007}, author = {Prakash, BM and Puttaraju, HP}, title = {Frequency of infection with A and B supergroup Wolbachia in insects and pests associated with mulberry and silkworm.}, journal = {Journal of biosciences}, volume = {32}, number = {4}, pages = {671-676}, pmid = {17762140}, issn = {0250-5991}, mesh = {Animals ; Base Sequence ; DNA Primers ; Insecta/*microbiology ; Polymerase Chain Reaction ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia is a ubiquitous, Gram-negative,vertically transmitted, alpha-proteobacterium that causes an array of reproductive abnormalities including cytoplasmic incompatibility, feminization of genetic males, parthenogenesis in a number of insect species, among others. Wolbachia is now being exploited as an agent for pest and vector control. Previous surveys indicated that it is commonly seen in 16-76% of arthropods. In this paper, using polymerase chain reaction assay based on specific amplification of the ftsZ -A and -B supergroup Wolbachia gene fragments, we found that 30% of insects and pests screened were positive for Wolbachia. Among them 66.7% harbour double Wolbachia infection, while 33.3 % harbour single Wolbachia infection. These results indicate widespread infection with both double and single Wolbachia, and provide a wealth of information to exploit this endobacterium for the management of pests and vectors.}, } @article {pmid17761848, year = {2007}, author = {Dunning Hotopp, JC and Clark, ME and Oliveira, DC and Foster, JM and Fischer, P and Muñoz Torres, MC and Giebel, JD and Kumar, N and Ishmael, N and Wang, S and Ingram, J and Nene, RV and Shepard, J and Tomkins, J and Richards, S and Spiro, DJ and Ghedin, E and Slatko, BE and Tettelin, H and Werren, JH}, title = {Widespread lateral gene transfer from intracellular bacteria to multicellular eukaryotes.}, journal = {Science (New York, N.Y.)}, volume = {317}, number = {5845}, pages = {1753-1756}, doi = {10.1126/science.1142490}, pmid = {17761848}, issn = {1095-9203}, mesh = {Animals ; Chromosome Mapping ; Crosses, Genetic ; DNA, Bacterial ; Drosophila/genetics/microbiology ; Female ; *Gene Transfer, Horizontal ; Genes, Bacterial ; In Situ Hybridization, Fluorescence ; Insecta/*genetics/microbiology ; Male ; Molecular Sequence Data ; Nematoda/*genetics/microbiology ; Retroelements ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {Although common among bacteria, lateral gene transfer-the movement of genes between distantly related organisms-is thought to occur only rarely between bacteria and multicellular eukaryotes. However, the presence of endosymbionts, such as Wolbachia pipientis, within some eukaryotic germlines may facilitate bacterial gene transfers to eukaryotic host genomes. We therefore examined host genomes for evidence of gene transfer events from Wolbachia bacteria to their hosts. We found and confirmed transfers into the genomes of four insect and four nematode species that range from nearly the entire Wolbachia genome (>1 megabase) to short (<500 base pairs) insertions. Potential Wolbachia-to-host transfers were also detected computationally in three additional sequenced insect genomes. We also show that some of these inserted Wolbachia genes are transcribed within eukaryotic cells lacking endosymbionts. Therefore, heritable lateral gene transfer occurs into eukaryotic hosts from their prokaryote symbionts, potentially providing a mechanism for acquisition of new genes and functions.}, } @article {pmid17727569, year = {2007}, author = {Daehnel, K and Gillette-Ferguson, I and Hise, AG and Diaconu, E and Harling, MJ and Heinzel, FP and Pearlman, E}, title = {Filaria/Wolbachia activation of dendritic cells and development of Th1-associated responses is dependent on Toll-like receptor 2 in a mouse model of ocular onchocerciasis (river blindness).}, journal = {Parasite immunology}, volume = {29}, number = {9}, pages = {455-465}, doi = {10.1111/j.1365-3024.2007.00962.x}, pmid = {17727569}, issn = {0141-9838}, support = {K08 AI054652/AI/NIAID NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Antibodies, Helminth/blood/immunology ; Antigens, CD/immunology/metabolism ; Brugia malayi/immunology ; Cornea/immunology ; Cytokines/immunology/metabolism ; Dendritic Cells/*immunology ; Disease Models, Animal ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Onchocerca volvulus/*immunology ; Onchocerciasis, Ocular/*immunology/metabolism ; Th1 Cells/*immunology ; Toll-Like Receptor 2/*immunology/metabolism ; Toll-Like Receptor 4/immunology/metabolism ; }, abstract = {Toll-like receptors (TLRs) regulate dendritic cell function and activate signals that mediate the nature of the adaptive immune response. The current study examined the role of TLRs in dendritic cell activation and in regulating T cell and antibody responses to antigens from the filarial parasites Onchocerca volvulus and Brugia malayi, which cause river blindness and lymphatic filariasis, respectively. Bone-marrow-derived CD11c(+) cells from C57BL/6 and TLR4(-/-) mice produced high levels of IL-6 and RANTES, and showed elevated surface CD40 expression, whereas CD11c(+) cells from myeloid differentiation factor 88(-/-) (MyD88(-/-)), TLR2(-/-) and TLR2/4(-/-) mice were not activated. Similarly, IFN-gamma production by splenocytes from immunized TLR2(-/-) mice was significantly impaired compared with splenocytes from C57BL/6 and TLR4(-/-) mice. In contrast, there was no difference among these strains in Th2-associated responses including IL-5 production by splenocytes from immunized animals, serum IgE and IgG(1), or eosinophil infiltration into the corneal stroma. Neutrophil recruitment to the cornea and CXC chemokine production was inhibited in immunized TLR2(-/-) mice compared with C57BL/6 and TLR4(-/-) mice. Taken together, these findings demonstrate an essential role for TLR2 in filaria-induced dendritic cell activation, IFN-gamma production and neutrophil migration to the cornea, but does not affect filaria-induced Th2-associated responses.}, } @article {pmid17693061, year = {2007}, author = {Riparbelli, MG and Giordano, R and Callaini, G}, title = {Effects of Wolbachia on sperm maturation and architecture in Drosophila simulans Riverside.}, journal = {Mechanisms of development}, volume = {124}, number = {9-10}, pages = {699-714}, doi = {10.1016/j.mod.2007.07.001}, pmid = {17693061}, issn = {0925-4773}, mesh = {Animals ; Cytoplasm/microbiology/physiology ; Drosophila/cytology/*microbiology ; Female ; Larva/growth & development/microbiology ; Male ; Pupa/microbiology ; Spermatozoa/cytology/*microbiology/pathology ; Testis/growth & development/microbiology ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is an intracellular obligate symbiont, that is relatively common in insects and also found in some nematodes. Cytoplasmic incompatibility (CI) is the most commonly expressed form, of several sex altering phenotypes caused by this rickettsial-like bacterium. CI is induced when infected males mate with uninfected females, and is likely the result of bacterial-induced modification of sperm grown in a Wolbachia-infected environment. Several studies have explored the dynamics of Wolbachia bacteria during sperm development in Drosophila. This study confirms and extends these earlier investigations of Wolbachia's distribution and proliferation in male germ cell lines. We examined Wolbachia population dynamics during testis development of Drosophila simulans (Riverside) by studying their distribution during the early mitotic divisions of secondary spermatogonial and subsequent meiotic cyst cells. Wolbachia are found in lower concentration in spermatogonial than in spermatocyte cells. Cytoplasmically incompatible crosses result in low levels of viable embryos despite the occurrence of fairly high levels of uninfected cysts. During meiotic divisions Wolbachia organize themselves at the poles during prophase and telophase but arrange themselves in equatorial bands during metaphase and anaphase. Moreover, during meiosis Wolbachia are asymmetrically divided between some daughter cells. There is no strong relationship between the fusome and Wolbachia and we have not found evidence that bacteria cross the ring canals. Wolbachia were observed at the distal and proximal sides of individualization complexes. Multiple altered sperm structures were observed during the process of individualization of infected sperm.}, } @article {pmid17688435, year = {2007}, author = {Lambros, S and Panos, I and Spiridon, L}, title = {Coding potential prediction in Wolbachia using artificial neural networks.}, journal = {In silico biology}, volume = {7}, number = {1}, pages = {105-113}, pmid = {17688435}, issn = {1386-6338}, mesh = {Algorithms ; Codon ; Computational Biology/*methods ; Gene Expression Profiling/*methods ; Genome, Bacterial ; Models, Statistical ; Models, Theoretical ; *Neural Networks, Computer ; Pattern Recognition, Automated ; Wolbachia/genetics/*metabolism ; }, abstract = {Ab initio coding potential prediction in a bacterial genome is an important step in determining an organism's transcriptional regulatory function. Extensive studies of genes structure have been carried out in a few species such as Escherichia coli, fewer resources exist in newly sequenced genomes like Wolbachia. A model of gene prediction trained on one species may not reflect the properties of other, distantly related prokaryotic organisms. These issues were encountered in the course of predicting genes in the genome of Wolbachia, very important gramnegative bacteria that form intracellular inherited infections in many invertebrates. We describe a coding potential predictor based on artificial neural networks and we compare its performance by using different architectures, learning algorithms and parameters. We rely on a dataset of positive samples constructed from coding sequences and on a negative dataset consisted of all the intergenic regions that were not located between the genes of an operon. Both datasets, positive and negative, were output as fasta formatted files and were used for neural network training. The fast, adaptive, batch learning algorithm Resilient propagation, exhibits the best overall performance on a 64input-10hidden-1output nodes multi-layer perceptron neural network.}, } @article {pmid17684565, year = {2007}, author = {Marshall, JL}, title = {Rapid evolution of spermathecal duct length in the Allonemobius socius complex of crickets: species, population and Wolbachia effects.}, journal = {PloS one}, volume = {2}, number = {8}, pages = {e720}, pmid = {17684565}, issn = {1932-6203}, mesh = {Animals ; *Biological Evolution ; Body Size ; Female ; Fertilization/physiology ; Genetic Speciation ; Genetic Variation ; Gonads/anatomy & histology ; Gryllidae/*anatomy & histology/classification/genetics/*microbiology ; Male ; Phylogeny ; Wolbachia/*physiology ; }, abstract = {The three species in the Allonemobius socius complex of crickets have recently diverged and radiated across North America. Interestingly, the only barriers to gene flow between these species in zones of secondary contact appear to be associated with fertilization traits - e.g., conspecific sperm precedence and the ability of males to induce females to lay eggs. Other traits, such as the length of female's reproductive tract, may also influence fertilization success and be associated with species boundaries. However, the underlying variation in this duct has not been assessed across populations and species. Moreover, the effects of reproductive parasites like Wolbachia on these morphological features have yet to be addressed, even though its infections are concentrated in reproductive tissues. I evaluated both the natural variation in and the effects of Wolbachia infection on spermathecal duct length among several populations of two species in the Allonemobius socius complex. My results suggest the following: (1) spermathecal duct length varies between species and is associated with species boundaries, (2) there is considerable variation among populations within species, (3) there is a Wolbachia infection-by-population interaction effect on the length of the spermathecal duct, and (4) experimental curing of Wolbachia recovers the uninfected morphology. These findings suggest the following hypotheses: (1) spermathecal duct length, like other fertilization traits in Allonemobius, is evolving rapidly and influences reproductive isolation and (2) Wolbachia-induced modifications of this duct could influence the dynamics of male-female coevolution. Further experiments are needed, however, to explicitly test these latter two hypotheses.}, } @article {pmid17684548, year = {2007}, author = {Telschow, A and Flor, M and Kobayashi, Y and Hammerstein, P and Werren, JH}, title = {Wolbachia-induced unidirectional cytoplasmic incompatibility and speciation: mainland-island model.}, journal = {PloS one}, volume = {2}, number = {8}, pages = {e701}, pmid = {17684548}, issn = {1932-6203}, mesh = {Animals ; *Cytoplasm ; Female ; Gene Flow ; *Genetic Speciation ; Genetics, Population ; *Geography ; Host-Parasite Interactions/genetics ; Insecta/genetics/microbiology ; Male ; *Models, Genetic ; Selection, Genetic ; Sexual Behavior, Animal ; Symbiosis/genetics ; *Wolbachia/cytology/genetics ; }, abstract = {Bacteria of the genus Wolbachia are among the most common endosymbionts in the world. In many insect species these bacteria induce a sperm-egg incompatibility between the gametes of infected males and uninfected females, commonly called unidirectional cytoplasmic incompatibility (CI). It is generally believed that unidirectional CI cannot promote speciation in hosts because infection differences between populations will be unstable and subsequent gene flow will eliminate genetic differences between diverging populations. In the present study we investigate this question theoretically in a mainland-island model with migration from mainland to island. Our analysis shows that (a) the infection polymorphism is stable below a critical migration rate, (b) an (initially) uninfected "island" can better maintain divergence at a selected locus (e.g. can adapt locally) in the presence of CI, and (c) unidirectional CI selects for premating isolation in (initially) uninfected island populations if they receive migration from a Wolbachia-infected mainland. Interestingly, premating isolation is most likely to evolve if levels of incompatibility are intermediate and if either the infection causes fecundity reductions or Wolbachia transmission is incomplete. This is because under these circumstances an infection pattern with an infected mainland and a mostly uninfected island can persist in the face of comparably high migration. We present analytical results for all three findings: (a) a lower estimation of the critical migration rate in the presence of local adaptation, (b) an analytical approximation for the gene flow reduction caused by unidirectional CI, and (c) a heuristic formula describing the invasion success of mutants at a mate preference locus. These findings generally suggest that Wolbachia-induced unidirectional CI can be a factor in divergence and speciation of hosts.}, } @article {pmid17684235, year = {2007}, author = {Pfarr, K and Foster, J and Slatko, B and Hoerauf, A and Eisen, JA}, title = {On the taxonomic status of the intracellular bacterium Wolbachia pipientis: should this species name include the intracellular bacteria of filarial nematodes?.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {57}, number = {Pt 8}, pages = {1677-1678}, pmid = {17684235}, issn = {1466-5026}, mesh = {Animals ; Filarioidea/*microbiology ; Wolbachia/*classification/genetics ; }, } @article {pmid17676427, year = {2007}, author = {Baldo, L and Prendini, L and Corthals, A and Werren, JH}, title = {Wolbachia are present in southern african scorpions and cluster with supergroup F.}, journal = {Current microbiology}, volume = {55}, number = {5}, pages = {367-373}, pmid = {17676427}, issn = {0343-8651}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/genetics ; Gene Transfer, Horizontal ; Genetic Variation ; Molecular Sequence Data ; *Phylogeny ; Scorpions/*microbiology ; Sequence Homology, Amino Acid ; South Africa ; Wolbachia/classification/*genetics/growth & development ; }, abstract = {The presence and distribution of the intracellular bacteria Wolbachia in the arthropod subphylum Chelicerata (including class Arachnida) has not been extensively explored. Here we report the discovery of Wolbachia in scorpions. Five strains found in host species of the genus Opistophthalmus (Southern African burrowing scorpions) have been characterized by Multilocus Sequence Typing and by Wolbachia Surface Protein. Phylogenetic analyses indicate clustering in the supergroup F and a high genetic relatedness among all scorpion strains as a result of a potential transmission within the host genus. The F-group is an uncommon lineage compared to the A and B supergroups, although it is present in a broad range of hosts (including insects, filarial nematodes, and now arachnids) and across a large geographical area (e.g., North America, Africa, Europe, and Australia). It also shows no evidence of recombination and has a significantly higher genetic diversity than supergroup A and B. Overall, this pattern suggests an older radiation of F-strains with respect to A and B-strains, followed by limited horizontal transmission across host genera and reduced genetic flux among strains. A more extensive sampling of supergroup F-strains is required to confirm this scenario.}, } @article {pmid17673369, year = {2007}, author = {Sréter-Lancz, Z and Széll, Z and Sréter, T}, title = {Molecular genetic comparison of Onchocerca sp. infecting dogs in Europe with other spirurid nematodes including Onchocerca lienalis.}, journal = {Veterinary parasitology}, volume = {148}, number = {3-4}, pages = {365-370}, doi = {10.1016/j.vetpar.2007.06.021}, pmid = {17673369}, issn = {0304-4017}, mesh = {Animals ; Dog Diseases/microbiology/*parasitology ; Dogs ; Electron Transport Complex IV/genetics ; Europe ; Female ; Molecular Sequence Data ; NADH Dehydrogenase/genetics ; Onchocerca/classification/enzymology/*genetics/microbiology ; Onchocerciasis, Ocular/microbiology/parasitology/*veterinary ; Phylogeny ; Spirurida/enzymology/*genetics ; Wolbachia/genetics ; }, abstract = {In the past 15 years, subconjunctival onchocercosis has been reported from 63 dogs in south-western United States (Arizona, California, Utah) and Southern and Central Europe (Germany, Greece, Hungary, Portugal, Switzerland). To reveal the taxonomic status of the parasite responsible for these infections, fragments of the mitochondrial cytochrome oxidase subunit I (COI) and NADH dehydrogenase subunit 5 (ND5) genes of three European strains of canine Onchocerca sp. and the 16S ribosomal RNA (16S rRNA) gene of their Wolbachia endosymbionts were sequenced and compared to the homologous sequences of other spirurid nematodes. The evolutionary divergence between COI and ND5 gene sequences of Greek, Hungarian and Portuguese strains of canine Onchocerca sp. were similar in magnitude to that seen within Thelazia callipaeda or Onchocerca lienalis. The evolutionary divergence between the sequences of canine Onchocerca sp. and other Onchocerca spp. including O. lienalis were similar or higher in magnitude to that seen between other Onchocerca spp. The results of the current and earlier phylogenetic analyses indicate that canine Onchocerca sp. separated from other Onchocerca spp. early in the evolution. Based on the similar clinical pictures, the identical morphology of nematodes and the sequence analyses of COI and ND5 genes of the worms and 16S rRNA gene of their wolbachiae, the Onchocerca worms isolated from European dogs appear to belong to the same species. The results support the earlier biological and morphological arguments that a distinct species, most likely O. lupi originally described from the subconjunctival tissues of a Caucasian wolf is responsible for canine ocular onchocercosis in Europe.}, } @article {pmid17663704, year = {2007}, author = {Roy, V and Harry, M}, title = {Diversity of Wolbachia isolated from the Cubitermes sp. affinis subarquatus complex of species (Termitidae), revealed by multigene phylogenies.}, journal = {FEMS microbiology letters}, volume = {274}, number = {1}, pages = {102-111}, doi = {10.1111/j.1574-6968.2007.00815.x}, pmid = {17663704}, issn = {0378-1097}, mesh = {Animals ; Bacterial Outer Membrane Proteins/chemistry/genetics ; Bacterial Proteins/chemistry/genetics ; Bacterial Typing Techniques ; Classification/methods ; Cytoskeletal Proteins/chemistry/genetics ; *Genes, Bacterial ; Isoptera/*microbiology ; *Phylogeny ; RNA, Ribosomal, 16S/chemistry ; Recombination, Genetic ; Sequence Analysis, DNA ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {Wolbachia are endosymbiotic bacteria that may alter the reproductive mechanisms of arthropod hosts. Eusocial termites provide considerable scope for Wolbachia studies owing to their ancient origin, their great diversity and their considerable ecological, biological and behavioral plasticity. This article describes the phylogenetic distribution of Wolbachia infecting termites of the Cubitermes genus, which are particularly abundant soil-feeders in equatorial Africa. Fourteen colonies of the Cubitermes sp. affinis subarquatus complex of species were screened using five bacterial genes (wsp, ftsZ, coxA, fbpA and 16S rRNA genes) and a striking diversity of Wolbachia strains was identified within these closely related species. In the host complex, three Wolbachia variants were found that were not in the super groups usually reported for termites (F and H), each infecting one or two Cubitermes species.}, } @article {pmid17660578, year = {2007}, author = {Yamada, R and Floate, KD and Riegler, M and O'Neill, SL}, title = {Male development time influences the strength of Wolbachia-induced cytoplasmic incompatibility expression in Drosophila melanogaster.}, journal = {Genetics}, volume = {177}, number = {2}, pages = {801-808}, pmid = {17660578}, issn = {0016-6731}, mesh = {Animals ; Cytoplasm/*physiology ; Drosophila melanogaster/growth & development/*microbiology ; Male ; Phenotype ; Symbiosis ; Testis/microbiology ; Wolbachia/*pathogenicity ; }, abstract = {Cytoplasmic incompatibility (CI) is the most widespread reproductive modification induced in insects by the maternally inherited intracellular bacteria, Wolbachia. Expression of CI in Drosophila melanogaster is quite variable. Published papers typically show that CI expression is weak and often varies between different Drosophila lines and different labs reporting the results. The basis for this variability is not well understood but is often considered to be due to unspecified host genotype interactions with Wolbachia. Here, we show that male development time can greatly influence CI expression in D. melanogaster. In a given family, males that develop fastest express very strong CI. The "younger brothers" of these males (males that take longer to undergo larval development) quickly lose their ability to express the CI phenotype as a function of development time. This effect is independent of male age effects and is enhanced when flies are reared under crowded conditions. No correlation is seen between this effect and Wolbachia densities in testes, suggesting that a more subtle interaction between host and symbiont is responsible. The observed younger brother effect may explain much of the reported variability in CI expression in this species. When male development time is controlled, it is possible to obtain consistently high levels of CI expression, which will benefit future studies that wish to use D. melanogaster as a model host to unravel CI mechanisms.}, } @article {pmid17657540, year = {2007}, author = {Duron, O and Gavotte, L}, title = {Absence of Wolbachia in nonfilariid worms parasitizing arthropods.}, journal = {Current microbiology}, volume = {55}, number = {3}, pages = {193-197}, pmid = {17657540}, issn = {0343-8651}, mesh = {Animals ; Biological Evolution ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; France ; Genes, Bacterial ; Helminths/*microbiology/physiology ; Insecta/physiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia are strictly intracellular maternally inherited alpha-proteobacteria, largely widespread among arthropods and filariids (i.e., filarial nematodes). Wolbachia capacities to infect new host species have been greatly evidenced and the transfer of Wolbachia between arthropods and filariids has probably occurred more than once. Interestingly, among nematode species, Wolbachia infection was found in filariids but not in closely related lineages. Their occurrence in filariids has been supposed a consequence of the parasitic lifestyle of worms within Wolbachia-infected arthropods, implying that nonfilariid worms parasitizing arthropods are also likely to be infected by some Wolbachia acquired from their hosts. To further investigate this hypothesis, we have examined seven species of nonfilariid worms of Nematoda and Nematomorpha phyla, all interacting intimately with arthropods. Wolbachia infection in nonfilariid parasitic worms was never detected by polymerase chain reaction assays of the 16S rDNA and wsp genes. By contrast, some arthropod hosts are well infected by Wolbachia of the B supergroup. Then the intimate contact with infected arthropods is not a sufficient condition to explain the Wolbachia occurrence in filariids and could underline a physiological singularity or a particular evolutionary event to acquire and maintain Wolbachia infection.}, } @article {pmid17645822, year = {2007}, author = {Chiel, E and Gottlieb, Y and Zchori-Fein, E and Mozes-Daube, N and Katzir, N and Inbar, M and Ghanim, M}, title = {Biotype-dependent secondary symbiont communities in sympatric populations of Bemisia tabaci.}, journal = {Bulletin of entomological research}, volume = {97}, number = {4}, pages = {407-413}, doi = {10.1017/S0007485307005159}, pmid = {17645822}, issn = {0007-4853}, mesh = {Animals ; Enterobacteriaceae/physiology ; Hemiptera/genetics/*microbiology/physiology ; Magnoliopsida/*parasitology ; Phenotype ; Rickettsia/physiology ; Symbiosis/*genetics/physiology ; Wolbachia/physiology ; }, abstract = {The sweet potato whitefly, Bemisia tabaci, harbors Portiera aleyrodidarum, an obligatory symbiotic bacterium, as well as several secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium and Fritschea, the function of which is unknown. Bemisia tabaci is a species complex composed of numerous biotypes, which may differ from each other both genetically and biologically. Only the B and Q biotypes have been reported from Israel. Secondary symbiont infection frequencies of Israeli laboratory and field populations of B. tabaci from various host plants were determined by PCR, in order to test for correlation between bacterial composition to biotype and host plant. Hamiltonella was detected only in populations of the B biotype, while Wolbachia and Arsenophonus were found only in the Q biotype (33% and 87% infection, respectively). Rickettsia was abundant in both biotypes. Cardinium and Fritschea were not found in any of the populations. No differences in secondary symbionts were found among host plants within the B biotype; but within the Q biotype, all whiteflies collected from sage harboured both Rickettsia and Arsenophonus, an infection frequency which was significantly higher than those found in association with all other host plants. The association found between whitefly biotypes and secondary symbionts suggests a possible contribution of these bacteria to host characteristics such as insecticide resistance, host range, virus transmission and speciation.}, } @article {pmid17642180, year = {2007}, author = {T/sillassie, H and Legesse, M}, title = {The role of Wolbachia bacteria in the pathogenesis of onchocerciasis and prospects for control of the disease.}, journal = {Ethiopian medical journal}, volume = {45}, number = {2}, pages = {213-219}, pmid = {17642180}, issn = {0014-1755}, mesh = {Humans ; Inflammation/microbiology ; Lipopolysaccharides/toxicity ; Onchocerciasis/drug therapy/*immunology/*microbiology ; Toll-Like Receptor 4/physiology ; Wolbachia/drug effects/*pathogenicity ; }, abstract = {Onchocerciasis, a non-fatal disease, is a major public health problem especially in sub-Saharan Africa causing disfigurement, severe itching, skin depigmentation, vision impairment and eventually blindness. The discovery of Wolbachia intracellular bacteria in the filarial nematodes has contributed a lot to the understanding of host's immune response to the bacteria and its role in the pathogenesis of onchocericiasis. Lipopolysaccharide molecules (LPS) associated with the bacteria are responsible for the induction of potent inflammatory responses mainly mediated by macrophages. LPS binding to CD14 on the monocytes/macrophages which is a co-receptor for Toll like receptor (TLR), induces the activation of an intracellular signaling that leads to the production of proinflammatory cytokines such as TNF-alpha, IL-1, IL-12 by macrophages. The cytokines initiate the recruitment of neturophils and macrophages to the vicinity of the adult worms/microfilaria especially to the cornea, which gradually causes corneal pathology. Therefore studies have suggested that Wolbachia plays an essential role in the induction of inflammatory response associated with the pathogenesis of onchocericiasis through the activation of innate immune response. Hence the aim of this study is to show the role wolbachia plays in the pathogenesis of this devastating illness and target for novel chemotherapy.}, } @article {pmid17641731, year = {2007}, author = {Porksakorn, C and Nuchprayoon, S and Park, K and Scott, AL}, title = {Proinflammatory cytokine gene expression by murine macrophages in response to Brugia malayi Wolbachia surface protein.}, journal = {Mediators of inflammation}, volume = {2007}, number = {}, pages = {84318}, pmid = {17641731}, issn = {1466-1861}, mesh = {Animals ; Bacterial Proteins/genetics/metabolism/*pharmacology ; Brugia malayi/microbiology ; Cell Line ; Cytokines/*genetics ; Gene Expression/*drug effects ; Interleukin-1beta/genetics ; Interleukin-6/genetics ; Macrophages/cytology/*drug effects/metabolism ; Mice ; RNA, Messenger/genetics/metabolism ; Recombinant Proteins/genetics/metabolism/*pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha/genetics ; Up-Regulation/drug effects/genetics ; Wolbachia/*metabolism ; }, abstract = {Wolbachia, an endosymbiotic bacterium found in most species of filarial parasites, is thought to play a significant role in inducing innate inflammatory responses in lymphatic filariasis patients. However, the Wolbachia-derived molecules that are recognized by the innate immune system have not yet been identified. In this study, we exposed the murine macrophage cell line RAW 264.7 to a recombinant form of the major Wolbachia surface protein (rWSP) to determine if WSP is capable of innately inducing cytokine transcription. Interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF) mRNAs were all upregulated by the rWSP stimulation in a dose-dependant manner. TNF transcription peaked at 3 hours, whereas IL-1beta and IL-6 transcription peaked at 6 hours post-rWSP exposure. The levels of innate cytokine expression induced by a high-dose (9.0 microg/mL) rWSP in the RAW 264.7 cells were comparable to the levels induced by 0.1 microg/mL E. coli-derived lipopolysaccharides. Pretreatment of the rWSP with proteinase-K drastically reduced IL-1beta, IL-6, and TNF transcription. However, the proinflammatory response was not inhibited by polymyxin B treatment. These results strongly suggest that the major Wolbachia surface protein molecule WSP is an important inducer of innate immune responses during filarial infections.}, } @article {pmid17626876, year = {2007}, author = {Charlat, S and Hornett, EA and Fullard, JH and Davies, N and Roderick, GK and Wedell, N and Hurst, GD}, title = {Extraordinary flux in sex ratio.}, journal = {Science (New York, N.Y.)}, volume = {317}, number = {5835}, pages = {214}, doi = {10.1126/science.1143369}, pmid = {17626876}, issn = {1095-9203}, mesh = {Animals ; Biological Evolution ; Butterflies/genetics/*microbiology/*physiology ; Female ; Genes, Insect ; Male ; Molecular Sequence Data ; Reproduction ; Samoa ; Selection, Genetic ; *Sex Ratio ; Wolbachia/genetics/*physiology ; }, abstract = {The ratio of males to females in a species is often considered to be relatively constant, at least over ecological time. Hamilton noted that the spread of "selfish" sex ratio-distorting elements could be rapid and produce a switch to highly biased population sex ratios. Selection against a highly skewed sex ratio should promote the spread of mutations that suppress the sex ratio distortion. We show that in the butterfly Hypolimnas bolina the suppression of sex biases occurs extremely fast, with a switch from a 100:1 population sex ratio to 1:1 occurring in fewer than 10 generations.}, } @article {pmid17621589, year = {2007}, author = {Monsen, KJ and Honchak, BM and Locke, SE and Peterson, MA}, title = {Cytonuclear disequilibrium in chrysochus hybrids is not due to patterns of mate choice.}, journal = {The Journal of heredity}, volume = {98}, number = {4}, pages = {325-330}, doi = {10.1093/jhered/esm039}, pmid = {17621589}, issn = {0022-1503}, mesh = {Animals ; Cell Nucleus/genetics ; Chimera/*genetics ; Choice Behavior/*physiology ; Coleoptera/*genetics/microbiology/physiology ; *Extrachromosomal Inheritance ; Female ; Genome, Insect ; Haplotypes ; *Linkage Disequilibrium ; Male ; *Pair Bond ; Wolbachia/genetics ; }, abstract = {We investigated patterns of cytonuclear disequilibrium between nuclear allozyme loci and partial mitochondrial COI and COII restriction fragment length polymorphism patterns within a population of hybridizing chrysomelid beetles and assessed to what degree the genotype frequencies of F1 hybrids were consistent with patterns of mate choice or endosymbiont infection. We document that in this population, > or = 50% of the heterospecific pairs at a given time are composed of Chrysochus auratus females and Chrysochus cobaltinus males, suggesting that at least half of the F1 hybrids should possess the C. auratus mitochondrial genotype. However, we found that the majority (89%) of F1 hybrids possessed C. cobaltinus mtDNA (P < 0.001). The lack of evidence for Wolbachia infection in these highly promiscuous beetles, coupled with the fact that F1 hybrids of both cross types do exist, indicates that endosymbionts are an unlikely explanation for the discrepancy between cytonuclear genotype frequencies and behavior. We argue that cytonuclear disequilibrium at this focal Chrysochus hybrid site is likely due to a strong directional bias in postmating prezygotic barriers in this system. The results presented here underscore the importance of combining both field and molecular data in studies of cytonuclear disequilibrium and point to the dangers inherent in attributing patterns of cytonuclear disequilibrium to assortative mating.}, } @article {pmid17588599, year = {2007}, author = {Reumer, BM and Kraaijeveld, K and van Alphen, JJ}, title = {Selection in the absence of males does not affect male-female conflict in the parasitoid wasp Leptopilina clavipes (Hymenoptera: Figitidae).}, journal = {Journal of insect physiology}, volume = {53}, number = {10}, pages = {994-999}, doi = {10.1016/j.jinsphys.2007.05.002}, pmid = {17588599}, issn = {0022-1910}, mesh = {Adaptation, Physiological/*physiology ; Animals ; Anti-Bacterial Agents/therapeutic use ; Female ; Longevity/physiology ; Male ; Reproduction/physiology ; Sexual Behavior, Animal/physiology ; Wasps/microbiology/*physiology ; Wolbachia/growth & development ; }, abstract = {Divergent reproductive interests of males and females can lead to sexually antagonistic coevolution (SAC). In the absence of males, adaptations evolved under SAC are released from selection and expected to deteriorate. In this study, we investigated this prediction using two populations of the parasitoid wasp Leptopilina clavipes, one arrhenotokous and one thelytokous. Thelytokous females were induced to produce sons by curing them of their Wolbachia-infection. We examined whether thelytokous males were less able to inhibit female remating than arrhenotokous males and whether thelytokous females were more susceptible to male-induced longevity reduction than arrhenotokous females. The results showed that females were monandrous, regardless of whether mated with an arrhenotokous or thelytokous male. While ongoing courtship of males reduced female life span, there was no longevity cost of mating for either arrhenotokous or thelytokous females. Our results therefore do not support the idea that adaptations evolved under SAC deteriorate under prolonged female-only selection.}, } @article {pmid17584494, year = {2007}, author = {Ioannidis, P and Dunning Hotopp, JC and Sapountzis, P and Siozios, S and Tsiamis, G and Bordenstein, SR and Baldo, L and Werren, JH and Bourtzis, K}, title = {New criteria for selecting the origin of DNA replication in Wolbachia and closely related bacteria.}, journal = {BMC genomics}, volume = {8}, number = {}, pages = {182}, pmid = {17584494}, issn = {1471-2164}, mesh = {Bacterial Proteins/metabolism ; Base Sequence ; Binding Sites ; *DNA Replication ; DNA-Binding Proteins/metabolism ; Integration Host Factors/metabolism ; *Replication Origin ; Transcription Factors/metabolism ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: The annotated genomes of two closely related strains of the intracellular bacterium Wolbachia pipientis have been reported without the identifications of the putative origin of replication (ori). Identifying the ori of these bacteria and related alpha-Proteobacteria as well as their patterns of sequence evolution will aid studies of cell replication and cell density, as well as the potential genetic manipulation of these widespread intracellular bacteria.

RESULTS: Using features that have been previously experimentally verified in the alpha-Proteobacterium Caulobacter crescentus, the origin of DNA replication (ori) regions were identified in silico for Wolbachia strains and eleven other related bacteria belonging to Ehrlichia, Anaplasma, and Rickettsia genera. These features include DnaA-, CtrA- and IHF-binding sites as well as the flanking genes in C. crescentus. The Wolbachia ori boundary genes were found to be hemE and COG1253 protein (CBS domain protein). Comparisons of the putative ori region among related Wolbachia strains showed higher conservation of bases within binding sites.

CONCLUSION: The sequences of the ori regions described here are only similar among closely related bacteria while fundamental characteristics like presence of DnaA and IHF binding sites as well as the boundary genes are more widely conserved. The relative paucity of CtrA binding sites in the ori regions, as well as the absence of key enzymes associated with DNA replication in the respective genomes, suggest that several of these obligate intracellular bacteria may have altered replication mechanisms. Based on these analyses, criteria are set forth for identifying the ori region in genome sequencing projects.}, } @article {pmid17571923, year = {2007}, author = {Toivonen, JM and Walker, GA and Martinez-Diaz, P and Bjedov, I and Driege, Y and Jacobs, HT and Gems, D and Partridge, L}, title = {No influence of Indy on lifespan in Drosophila after correction for genetic and cytoplasmic background effects.}, journal = {PLoS genetics}, volume = {3}, number = {6}, pages = {e95}, pmid = {17571923}, issn = {1553-7404}, support = {/WT_/Wellcome Trust/United Kingdom ; BB/C51705X/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; SF19106/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Caenorhabditis elegans/genetics/physiology ; Cytoplasm/genetics/*physiology ; DNA, Mitochondrial/physiology ; Dicarboxylic Acid Transporters/genetics/*physiology ; Drosophila/*genetics/growth & development/physiology ; Drosophila Proteins/genetics/*physiology ; Female ; Longevity/*genetics/physiology ; Male ; Mitochondria/genetics/physiology ; Molecular Sequence Data ; Mutation ; Symporters/genetics/*physiology ; }, abstract = {To investigate whether alterations in mitochondrial metabolism affect longevity in Drosophila melanogaster, we studied lifespan in various single gene mutants, using inbred and outbred genetic backgrounds. As positive controls we included the two most intensively studied mutants of Indy, which encodes a Drosophila Krebs cycle intermediate transporter. It has been reported that flies heterozygous for these Indy mutations, which lie outside the coding region, show almost a doubling of lifespan. We report that only one of the two mutants lowers mRNA levels, implying that the lifespan extension observed is not attributable to the Indy mutations themselves. Moreover, neither Indy mutation extended lifespan in female flies in any genetic background tested. In the original genetic background, only the Indy mutation associated with altered RNA expression extended lifespan in male flies. However, this effect was abolished by backcrossing into standard outbred genetic backgrounds, and was associated with an unidentified locus on the X chromosome. The original Indy line with long-lived males is infected by the cytoplasmic symbiont Wolbachia, and the longevity of Indy males disappeared after tetracycline clearance of this endosymbiont. These findings underscore the critical importance of standardisation of genetic background and of cytoplasm in genetic studies of lifespan, and show that the lifespan extension previously claimed for Indy mutants was entirely attributable to confounding variation from these two sources. In addition, we saw no effects on lifespan of expression knockdown of the Indy orthologues nac-2 and nac-3 in the nematode Caenorhabditis elegans.}, } @article {pmid17562224, year = {2007}, author = {Tsai, KH and Huang, CG and Wang, LC and Yu, YW and Wu, WJ and Chen, WJ}, title = {Molecular evidence for the endosymbiont Wolbachia in a non-filaroid nematode, Angiostrongylus cantonensis.}, journal = {Journal of biomedical science}, volume = {14}, number = {5}, pages = {607-615}, doi = {10.1007/s11373-007-9181-3}, pmid = {17562224}, issn = {1021-7770}, mesh = {Angiostrongylus cantonensis/*microbiology ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Base Sequence ; Cytoskeletal Proteins/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Wolbachia harbored by most filarial parasites, is critical to both embryogenesis and microfilarial development, and may lead to inflammation and pathogenesis in infected hosts. Based on alignment of the sequences from the wsp, ftsZ, and 16S rRNA genes, Wolbachia was demonstrated to exist in Angiostrongylus cantonensis, a non-filaroid nematode. Although the wsp gene may not be the best candidate for evolutionary analysis of Wolbachia, this gene has been sequenced from a broader coverage of the host species, making it feasible to be used for phylogenetic analysis in this study. The results from both Neighbor-joining and Maximum parsimony methods showed that this novel Wolbachia does not belong to any of the known groups (C or D) of nematode-derived Wolbachia. In addition, the wsp gene sequence of this newly identified endosymbiont revealed a high degree of identity (98%) with that from Diaea circumlita c2, tentatively classified into the putative group G. This suggests that Wolbachia from A. cantonensis could represent a deeply branched lineage in Wolbachia evolution or the occurrence of horizontal transfer between infected hosts. In conclusion, the findings provide some insights into our understanding of the evolution of Wolbachia, particularly the isolate from A. cantonensis.}, } @article {pmid17554631, year = {2007}, author = {Enigl, M and Schausberger, P}, title = {Incidence of the endosymbionts Wolbachia, Cardinium and Spiroplasma in phytoseiid mites and associated prey.}, journal = {Experimental & applied acarology}, volume = {42}, number = {2}, pages = {75-85}, pmid = {17554631}, issn = {0168-8162}, mesh = {Animals ; Bacteroidetes/genetics/*growth & development ; DNA, Bacterial/chemistry/genetics ; Incidence ; Mites/*microbiology ; Pest Control, Biological/methods ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Spiroplasma/genetics/*growth & development ; Symbiosis ; Wolbachia/genetics/*growth & development ; }, abstract = {Endosymbiotic bacteria that potentially influence reproduction and other fitness-related traits of their hosts are widespread in insects and mites and their appeal to researchers' interest is still increasing. We screened 20 strains of 12 agriculturally relevant herbivorous and predatory mite species for infection with Wolbachia, Cardinium and Spiroplasma by the use of PCR. The majority of specimens originated from Austria and were field collected or mass-reared. Eight out of 20 strains (40%) tested, representing seven of 12 mite species (58%), carried at least one of the three bacteria. We found Wolbachia in the herbivorous spider mites Tetranychus urticae and Bryobia rubrioculus, with the former also carrying Spiroplasma and the latter also carrying Cardinium. Cardinium was furthermore found in two populations of the predatory mite Euseius finlandicus and the spider mite Eotetranychus uncatus. Spiroplasma was detected in the predatory mite Neoseiulus californicus. All bacteria positive PCR products were sequenced, submitted to GenBank and analyzed in BLAST queries. We found high similarities to complete identity with bacteria found in the same and different mite species but also with bacteria found in insect species like ladybirds, butterflies and minute pirate bugs, Orius. We discuss the significance of potential (multiple) infections with the investigated bacteria for biological control.}, } @article {pmid17551786, year = {2007}, author = {Baldo, L and Werren, JH}, title = {Revisiting Wolbachia supergroup typing based on WSP: spurious lineages and discordance with MLST.}, journal = {Current microbiology}, volume = {55}, number = {1}, pages = {81-87}, pmid = {17551786}, issn = {0343-8651}, mesh = {Bacterial Outer Membrane Proteins/*genetics ; Bacterial Typing Techniques/*methods ; DNA, Bacterial/genetics ; Genotype ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology ; Wolbachia/*classification/*genetics ; }, abstract = {The obligate intracellular bacteria Wolbachia are taxonomically subdivided into eight supergroups (named A-H). Supergroup typing of strains has been mostly based on phylogenetic inference of the Wolbachia surface protein (wsp), a gene that recently has been shown to experience high rates of recombination. This brings into question its suitability not only for microtaxonomy, but also for supergroup classification of the genus. A Multilocus Sequence Typing (MLST) scheme for Wolbachia has recently been developed that types strains based on five conserved genes, thus providing a rigorous supergroup annotation of strains. Here we report striking discrepancies in supergroup designation between MLST and wsp inferences, and propose a revision of current methods for Wolbachia supergroup typing. Transfer of whole wsp gene sequences between supergroups A and B has occurred. Furthermore, as a result of intragenic recombination, wsp phylogeny creates spurious basal lineages that are not supported by MLST. For example, the proposed supergroup G, based upon wsp alone, likely represents only a wsp recombinant clade. Removal of supergroup G is advised until and unless the existence of this lineage is substantiated by other sequence information (e.g., MLST). We recommend a full characterization MLST for a correct strain typing, while, based on the current data set, use of a single MLST gene can be effective for supergroup designation of A and B strains. Finally, we note that the sharing of wsp sequences between A and B strains indicates a strong genetic cohesiveness of Wolbachia strains, supporting designation of these bacteria within the same species, W. pipientis.}, } @article {pmid17550649, year = {2007}, author = {Kassem, HA and Osman, G}, title = {Maternal transmission of Wolbachia in Phlebotomus papatasi (Scopoli).}, journal = {Annals of tropical medicine and parasitology}, volume = {101}, number = {5}, pages = {435-440}, doi = {10.1179/136485913X13789813917544}, pmid = {17550649}, issn = {0003-4983}, mesh = {Animals ; Cytoplasm ; Female ; *Infectious Disease Transmission, Vertical ; Insect Vectors/*parasitology ; Male ; Phlebotomus/*parasitology ; Polymerase Chain Reaction/methods ; Rickettsiaceae Infections/*transmission ; *Wolbachia ; }, abstract = {Attempts have been made to transfer Wolbachia from infected to uninfected, laboratory-reared Phlebotomus papatasi, through mating, and to determine whether the incompatibility phenotype could be expressed through crosses between infected and uninfected flies. In order to test for the intraspecific transmission of Wolbachia in crosses between infected females and uninfected males, or those between uninfected females and infected males, a PCR based on Wolbachia -specific wsp primers was used to test the progeny of each cross and, subsequently, 50 individual flies from the F(3) generation. All the individual flies tested from the F(1) progeny of the crosses between infected males and uninfected females were found to be uninfected. In the crosses involving infected females and uninfected males, however, Wolbachia were found in the progeny of five matings out of the 23 that produced viable eggs. In the F(3), Wolbachia were not detected in any of the individuals resulting from the cross between uninfected females and infected males but they were detected in 52% (26) of the 50 tested individuals resulting from the cross between infected females and uninfected males. No evidence of cytoplasmic incompatibility (CI) was observed in any of the crosses. The absence of CI expression and relatively low frequencies of maternal transmission could hamper the potential use of Wolbachia in a transgenic strategy for the control of leishmaniases.}, } @article {pmid17546095, year = {2007}, author = {Narita, S and Nomura, M and Kageyama, D}, title = {A natural population of the butterfly Eurema hecabe with Wolbachia-induced female-biased sex ratio not by feminization.}, journal = {Genome}, volume = {50}, number = {4}, pages = {365-372}, doi = {10.1139/g07-020}, pmid = {17546095}, issn = {0831-2796}, mesh = {Animals ; Butterflies/*genetics/*microbiology ; Female ; Feminization/microbiology ; Male ; *Sex Determination Processes ; *Sex Ratio ; Wolbachia/*physiology ; }, abstract = {In butterflies, the adult sex ratio observed in the field is usually male-biased, although the sex ratio of their progeny is 1:1. This is due to the higher motility and larger behavioral range of males than females. As expected, the sex ratio of Eurema hecabe butterflies collected at 6 localities throughout Japan was male-biased. However, in Tsukuba, located in the central part of Japan, the sex ratio was found to be biased toward females. Their progeny reared in the laboratory also exhibited a female-biased sex ratio. A single strain of Wolbachia is considered to be the cause of the sex-ratio distortion, because antibiotic treatment reversed the sex ratio to 1:1, and only a single nucleotide sequence of wsp, a highly variable Wolbachia gene, was detected by molecular analysis. Cytogenetic analysis excluded the possibility of feminization as the underlying mechanism. In addition, when the wild-caught females that had already mated in nature were treated with antibiotics before oviposition, egg-hatch rates were extremely low, suggesting that the same Wolbachia strain also caused cytoplasmic incompatibility. Our findings suggest the possibility that a single strain of Wolbachia induces 2 distinct reproductive manipulations in the same host.}, } @article {pmid17537669, year = {2007}, author = {Iturbe-Ormaetxe, I and O'Neill, SL}, title = {Wolbachia-host interactions: connecting phenotype to genotype.}, journal = {Current opinion in microbiology}, volume = {10}, number = {3}, pages = {221-224}, doi = {10.1016/j.mib.2007.05.002}, pmid = {17537669}, issn = {1369-5274}, mesh = {Animals ; *Genome, Bacterial ; Genomics/methods ; Genotype ; Insecta/microbiology ; Phenotype ; Wolbachia/*genetics/growth & development/metabolism ; }, abstract = {The long-established view of Wolbachia as reproductive parasites of insects is becoming complicated as an increasing number of papers describe a richer picture of Wolbachia-mediated phenotypes in insects. The search for the molecular basis for this phenotypic variability has been greatly aided by the recent sequencing of several Wolbachia genomes. These studies have revealed putative genes and pathways that are likely to be involved in the host-symbiont interaction. Whereas significant progress is being made from comparative genomic studies together with the use of model host systems like Drosophila, the ultimate linking of phenotype to genotype will require the development of genetic manipulation technology for both host and symbiont.}, } @article {pmid17537593, year = {2007}, author = {Verne, S and Johnson, M and Bouchon, D and Grandjean, F}, title = {Evidence for recombination between feminizing Wolbachia in the isopod genus Armadillidium.}, journal = {Gene}, volume = {397}, number = {1-2}, pages = {58-66}, doi = {10.1016/j.gene.2007.04.006}, pmid = {17537593}, issn = {0378-1119}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; DNA, Bacterial/genetics ; Evolution, Molecular ; Female ; Feminization ; Genes, Bacterial ; Genetic Variation ; Isopoda/*microbiology ; Male ; Molecular Sequence Data ; Phylogeny ; Polymorphism, Genetic ; *Recombination, Genetic ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid ; Species Specificity ; Symbiosis/genetics ; Wolbachia/classification/*genetics/isolation & purification/physiology ; }, abstract = {Wolbachia are maternally inherited endosymbiotic alpha-Proteobacteria infecting a wide range of arthropods. Wolbachia induce feminization in many terrestrial isopod species, particularly in the genus Armadillidium (Crustacea, Oniscidea). The diversity of Wolbachia strains infecting Armadillidium species was examined. Results reveal that natural populations of A. vulgare contain three different Wolbachia strains (wVulC, wVulM and wVulP). The wsp gene and its 3'-adjacent region show evidence that two recombination events have occurred between two of these strains. In both cases, multiple statistical analyses suggest that a small gene fragment of a strain closely related to wVulM (minor parent) is inserted into the genome of another strain closely related to wVulC (major parent). Although multiple infections in a single individual have never been demonstrated in natural population, the existence of recombination between feminizing strains suggests that bi-infections are possible, or at least that bi-infections can be maintained sufficiently long enough to allow recombination. Recombination events increase genetic diversity of Wolbachia found in Armadillidium species and may play a role in the ability of Wolbachia strains to invade new hosts.}, } @article {pmid17519968, year = {2007}, author = {Bordenstein, SR and Werren, JH}, title = {Bidirectional incompatibility among divergent Wolbachia and incompatibility level differences among closely related Wolbachia in Nasonia.}, journal = {Heredity}, volume = {99}, number = {3}, pages = {278-287}, doi = {10.1038/sj.hdy.6800994}, pmid = {17519968}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Genome, Bacterial/genetics ; Genome, Insect/genetics ; Species Specificity ; Symbiosis/*genetics ; Wasps/*genetics/*microbiology ; Wolbachia/*genetics ; }, abstract = {Most insect groups harbor obligate bacterial symbionts from the alpha-proteobacterial genus Wolbachia. These bacteria alter insect reproduction in ways that enhance their cytoplasmic transmission. One of the most common alterations is cytoplasmic incompatibility (CI) - a post-fertilization modification of the paternal genome that renders embryos inviable or unable to complete diploid development in crosses between infected males and uninfected females or infected females harboring a different strain. The parasitic wasp species complex Nasonia (N. vitripennis, N. longicornis and N. giraulti) harbor at least six different Wolbachia that cause CI. Each species have double infections with a representative from both the A and B Wolbachia subgroups. CI relationships of the A and B Wolbachia of N. longicornis with those of N. giraulti and N. vitripennis are investigated here. We demonstrate that all pairwise crosses between the divergent A strains are bidirectionally incompatible. We were unable to characterize incompatibility between the B Wolbachia, but we establish that the B strain of N. longicornis induces no or very weak CI in comparison to the closely related B strain in N. giraulti that expresses complete CI. Taken together with previous studies, we show that independent acquisition of divergent A Wolbachia has resulted in three mutually incompatible strains, whereas codivergence of B Wolbachia in N. longicornis and N. giraulti is associated with differences in CI level. Understanding the diversity and evolution of new incompatibility strains will contribute to a fuller understanding of Wolbachia invasion dynamics and Wolbachia-assisted speciation in certain groups of insects.}, } @article {pmid17519957, year = {2007}, author = {Duron, O and Fort, P and Weill, M}, title = {Influence of aging on cytoplasmic incompatibility, sperm modification and Wolbachia density in Culex pipiens mosquitoes.}, journal = {Heredity}, volume = {98}, number = {6}, pages = {368-374}, doi = {10.1038/sj.hdy.6800948}, pmid = {17519957}, issn = {0018-067X}, mesh = {Aging/*physiology ; Animals ; Colony Count, Microbial ; Crosses, Genetic ; Culex/*microbiology ; *Extrachromosomal Inheritance ; Female ; Male ; Spermatogenesis/*physiology ; Testis/microbiology/physiology ; Wolbachia/*growth & development ; }, abstract = {Wolbachia are maternally inherited endocellular bacteria, widespread in invertebrates and capable of altering several aspects of host reproduction. Cytoplasmic incompatibility (CI) is commonly found in arthropods and induces hatching failure of eggs from crosses between Wolbachia-infected males and uninfected females (or females infected by incompatible strains). Several factors such as bacterial and host genotypes or bacterial density contribute to CI strength and it has been proposed, mostly from Drosophila data, that older males have a lower Wolbachia load in testes which, thus, induces a lighter CI. Here, we challenge this hypothesis using different incompatible Culex pipiens mosquito strains and show that CI persists at the same intensity throughout the mosquito life span. Embryos from incompatible crosses showed even distributions of abortive phenotypes over time, suggesting that host ageing does not reduce the sperm-modification induced by Wolbachia. CI remained constant when sperm was placed in the spermathecae of incompatible females, indicating that sperm modification is also stable over time. The capacity of infected females to rescue CI was independent of age. Last, the density of Wolbachia in whole testes was highly strain-dependent and increased dramatically with age. Taken together, these data stress the peculiarity of the C.pipiens/Wolbachia interaction and suggest that the bacterial dosage model should be rejected in the case of this association.}, } @article {pmid17506822, year = {2007}, author = {Narita, S and Nomura, M and Kageyama, D}, title = {Naturally occurring single and double infection with Wolbachia strains in the butterfly Eurema hecabe: transmission efficiencies and population density dynamics of each Wolbachia strain.}, journal = {FEMS microbiology ecology}, volume = {61}, number = {2}, pages = {235-245}, doi = {10.1111/j.1574-6941.2007.00333.x}, pmid = {17506822}, issn = {0168-6496}, mesh = {Animals ; Butterflies/embryology/growth & development/*microbiology ; Colony Count, Microbial ; Embryonic Development ; Female ; Infectious Disease Transmission, Vertical ; Larva/growth & development/microbiology ; Male ; Mortality ; Pupa/growth & development/microbiology ; Sex Ratio ; Wolbachia/classification/isolation & purification/*physiology ; }, abstract = {Wolbachia belonging to Alphaproteobacteria are transovarially transmitted bacteria responsible for reproductive alterations in a wide range of arthropods. In natural populations of the butterfly Eurema hecabe, there are two different types of Wolbachia-infected individuals. Individuals singly infected with Wolbachia strain wHecCI exhibit strong cytoplasmic incompatibility, whereas those doubly infected with wHecCI and wHecFem exhibit feminization. Here, we examined the infection frequencies and population densities of each Wolbachia strain in different host tissues (ovary, testis, fat body, midgut, Malpighian tubule and leg), and the cost of infection in offspring produced by single-infected and double-infected mothers of E. hecabe. The vertical transmission rate of wHecCI was nearly 100%, and that of wHecFem was c. 80%. The wHecCI densities were 10(3)-10(4)-fold higher than the wHecFem densities. In most tissues, the wHecCI densities were significantly higher in offspring of single-infected mothers than in offspring of double-infected mothers. In offspring of double-infected mothers, however, the wHecCI densities were not affected by the presence of wHecFem, suggesting a lack of interaction between the wHecCI and wHecFem densities. The offspring development time was dependent on the infection status of the mothers. These results imply that the maternal infection status affects the Wolbachia densities and fitness of offspring.}, } @article {pmid17496135, year = {2007}, author = {Narita, S and Kageyama, D and Nomura, M and Fukatsu, T}, title = {Unexpected mechanism of symbiont-induced reversal of insect sex: feminizing Wolbachia continuously acts on the butterfly Eurema hecabe during larval development.}, journal = {Applied and environmental microbiology}, volume = {73}, number = {13}, pages = {4332-4341}, pmid = {17496135}, issn = {0099-2240}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Base Sequence ; Colony Count, Microbial ; Crustacea/growth & development/microbiology ; DNA Primers/genetics ; DNA, Bacterial/genetics ; Disorders of Sex Development ; Female ; Feminization ; Insecta/growth & development/microbiology ; Larva/growth & development/microbiology ; Male ; Mosaicism ; Moths/drug effects/*growth & development/*microbiology ; Phenotype ; Symbiosis/*physiology ; Wings, Animal/growth & development ; Wolbachia/drug effects/genetics/isolation & purification/*physiology ; }, abstract = {When the butterfly Eurema hecabe is infected with two different strains (wHecCI2 and wHecFem2) of the bacterial endosymbiont Wolbachia, genetic males are transformed into functional females, resulting in production of all-female broods. In an attempt to understand how and when the Wolbachia endosymbiont feminizes genetically male insects, larval insects were fed an antibiotic-containing diet beginning at different developmental stages until pupation. When the adult insects emerged, strikingly, many of them exhibited sexually intermediate traits in their wings, reproductive organs, and genitalia. The expression of intersexual phenotypes was strong in the insects treated from first instar, moderate in the insects treated from third instar, and weak in the insects treated from fourth instar. The insects treated from early larval instar grew and pupated normally but frequently failed to emerge and died in the pupal case. The dead insects in the pupal case contained lower densities of the feminizing Wolbachia endosymbiont than the successfully emerged insects, although none of them were completely cured of the symbiont infection. These results suggest the following: (i) the antibiotic treatment suppressed the population of feminizing Wolbachia endosymbionts; (ii) the suppression probably resulted in attenuated feminizing activity of the symbiont, leading to expression of intersexual host traits; (iii) many of the insects suffered pupal mortality, possibly due to either intersexual defects or Wolbachia-mediated addiction; and hence (iv) the feminizing Wolbachia endosymbiont continuously acts on the host insects during larval development for expression of female phenotypes under a male genotype. Our finding may prompt reconsideration of the notion that Wolbachia-induced reproductive manipulations are already complete before the early embryonic stage and provide insights into the mechanism underlying the symbiont-induced reversal of insect sex.}, } @article {pmid17487529, year = {2007}, author = {Li, ZX and Lin, HZ and Guo, XP}, title = {Prevalence of Wolbachia infection in Bemisia tabaci.}, journal = {Current microbiology}, volume = {54}, number = {6}, pages = {467-471}, pmid = {17487529}, issn = {0343-8651}, mesh = {Animals ; Hemiptera/*microbiology ; Molecular Sequence Data ; Phylogeny ; Polymorphism, Restriction Fragment Length ; Wolbachia/classification/*genetics/pathogenicity ; }, abstract = {Wolbachia are obligate intracellular bacteria present in reproductive tissues of many arthropod species. It has been reported that few silverleafing populations of Bemisia tabaci were positive for Wolbachia, whereas non-silverleafing populations were more likely infected with Wolbachia and all that infect B. tabaci are Wolbachia belonging to supergroup B. However, current detection methods were shown to be not sensitive enough to uncover all infections. Herein, a protocol based on polymerase chain reaction-restriction fragment length polymorphism analysis of Wolbachia 16S ribosomal DNA is presented. A systematic survey for the prevalence of Wolbachia infection in natural populations of B. tabaci using this method revealed that (1) all populations of B. tabaci tested positive for Wolbachia and the overall infection rate reached 80.5% (293 positives in 364 tests); (2) both single infection and superinfection existed within individual whiteflies tested; and (3) silverleafing populations of B. tabaci most likely harbored A Wolbachia as single infection, whereas non-silverleafing populations tend to carry B Wolbachia as superinfection. It is clear that the Wolbachia infection pattern is closely related to the genetic races of B. tabaci, and the infection frequencies are apparently much higher than those described previously. This study shows that detection methods can significantly influence estimation of Wolbachia infection. It is supposed that Wolbachia may be acting as a biotic agent promoting rapid differentiation and speciation of B. tabaci. This is the most systematic survey of Wolbachia infection within B. tabaci.}, } @article {pmid17472911, year = {2007}, author = {Whitworth, TL and Dawson, RD and Magalon, H and Baudry, E}, title = {DNA barcoding cannot reliably identify species of the blowfly genus Protocalliphora (Diptera: Calliphoridae).}, journal = {Proceedings. Biological sciences}, volume = {274}, number = {1619}, pages = {1731-1739}, pmid = {17472911}, issn = {0962-8452}, mesh = {Animals ; Classification/*methods ; Diptera/classification/*genetics/microbiology ; Electron Transport Complex IV/genetics ; *Genetic Variation ; Hybridization, Genetic ; Likelihood Functions ; Models, Genetic ; *Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; United States ; Wolbachia/genetics ; }, abstract = {In DNA barcoding, a short standardized DNA sequence is used to assign unknown individuals to species and aid in the discovery of new species. A fragment of the mitochondrial gene cytochrome c oxidase subunit 1 is emerging as the standard barcode region for animals. However, patterns of mitochondrial variability can be confounded by the spread of maternally transmitted bacteria that cosegregate with mitochondria. Here, we investigated the performance of barcoding in a sample comprising 12 species of the blow fly genus Protocalliphora, known to be infected with the endosymbiotic bacteria Wolbachia. We found that the barcoding approach showed very limited success: assignment of unknown individuals to species is impossible for 60% of the species, while using the technique to identify new species would underestimate the species number in the genus by 75%. This very low success of the barcoding approach is due to the non-monophyly of many of the species at the mitochondrial level. We even observed individuals from four different species with identical barcodes, which is, to our knowledge, the most extensive case of mtDNA haplotype sharing yet described. The pattern of Wolbachia infection strongly suggests that the lack of within-species monophyly results from introgressive hybridization associated with Wolbachia infection. Given that Wolbachia is known to infect between 15 and 75% of insect species, we conclude that identification at the species level based on mitochondrial sequence might not be possible for many insects. However, given that Wolbachia-associated mtDNA introgression is probably limited to very closely related species, identification at the genus level should remain possible.}, } @article {pmid17470297, year = {2007}, author = {Brownlie, JC and Adamski, M and Slatko, B and McGraw, EA}, title = {Diversifying selection and host adaptation in two endosymbiont genomes.}, journal = {BMC evolutionary biology}, volume = {7}, number = {}, pages = {68}, pmid = {17470297}, issn = {1471-2148}, mesh = {*Adaptation, Physiological ; Animals ; Brugia malayi/genetics/*microbiology ; Drosophila melanogaster/genetics/*microbiology ; Genome ; Host-Parasite Interactions ; *Selection, Genetic ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: The endosymbiont Wolbachia pipientis infects a broad range of arthropod and filarial nematode hosts. These diverse associations form an attractive model for understanding host:symbiont coevolution. Wolbachia's ubiquity and ability to dramatically alter host reproductive biology also form the foundation of research strategies aimed at controlling insect pests and vector-borne disease. The Wolbachia strains that infect nematodes are phylogenetically distinct, strictly vertically transmitted, and required by their hosts for growth and reproduction. Insects in contrast form more fluid associations with Wolbachia. In these taxa, host populations are most often polymorphic for infection, horizontal transmission occurs between distantly related hosts, and direct fitness effects on hosts are mild. Despite extensive interest in the Wolbachia system for many years, relatively little is known about the molecular mechanisms that mediate its varied interactions with different hosts. We have compared the genomes of the Wolbachia that infect Drosophila melanogaster, wMel and the nematode Brugia malayi, wBm to that of an outgroup Anaplasma marginale to identify genes that have experienced diversifying selection in the Wolbachia lineages. The goal of the study was to identify likely molecular mechanisms of the symbiosis and to understand the nature of the diverse association across different hosts.

RESULTS: The prevalence of selection was far greater in wMel than wBm. Genes contributing to DNA metabolism, cofactor biosynthesis, and secretion were positively selected in both lineages. In wMel there was a greater emphasis on DNA repair, cell division, protein stability, and cell envelope synthesis.

CONCLUSION: Secretion pathways and outer surface protein encoding genes are highly affected by selection in keeping with host:parasite theory. If evidence of selection on various cofactor molecules reflects possible provisioning, then both insect as well as nematode Wolbachia may be providing substances to hosts. Selection on cell envelope synthesis, DNA replication and repair machinery, heat shock, and two component switching suggest strategies insect Wolbachia may employ to cope with diverse host and intra-host environments.}, } @article {pmid17449622, year = {2007}, author = {Duron, O and Boureux, A and Echaubard, P and Berthomieu, A and Berticat, C and Fort, P and Weill, M}, title = {Variability and expression of ankyrin domain genes in Wolbachia variants infecting the mosquito Culex pipiens.}, journal = {Journal of bacteriology}, volume = {189}, number = {12}, pages = {4442-4448}, pmid = {17449622}, issn = {0021-9193}, mesh = {Amino Acid Motifs ; Animals ; Ankyrins/chemistry/*genetics ; Bacterial Proteins/chemistry/*genetics ; Crosses, Genetic ; Culex/*microbiology ; DNA, Bacterial/chemistry/genetics ; Female ; Gene Expression ; Male ; Molecular Sequence Data ; Polymorphism, Genetic ; Protein Structure, Tertiary ; Sequence Analysis, DNA ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {Wolbachia strains are maternally inherited endosymbiotic bacteria that infect many arthropod species and have evolved several different ways of manipulating their hosts, the most frequent way being cytoplasmic incompatibility (CI). CI leads to embryo death in crosses between infected males and uninfected females as well as in crosses between individuals infected by incompatible Wolbachia strains. The mosquito Culex pipiens exhibits the highest crossing type variability reported so far. Our crossing data support the notion that CI might be driven by at least two distinct genetic units that control the CI functions independently in males and females. Although the molecular basis of CI remains unknown, proteins with ankyrin (ANK) domains represent promising candidates since they might interact with a wide range of host proteins. Here we searched for sequence variability in the 58 ANK genes carried in the genomes of Wolbachia variants infecting Culex pipiens. Only five ANK genes were polymorphic in the genomes of incompatible Wolbachia variants, and none correlated with the CI pattern obtained with 15 mosquito strains (representing 14 Wolbachia variants). Further analysis of ANK gene expression evidenced host- and sex-dependent variations, which did not improve the correlation. Taken together, these data do not support the direct implication of ANK genes in CI determinism.}, } @article {pmid17447012, year = {2007}, author = {Gotoh, T and Sugasawa, J and Noda, H and Kitashima, Y}, title = {Wolbachia-induced cytoplasmic incompatibility in Japanese populations of Tetranychus urticae (Acari: Tetranychidae).}, journal = {Experimental & applied acarology}, volume = {42}, number = {1}, pages = {1-16}, pmid = {17447012}, issn = {0168-8162}, mesh = {Animals ; Breeding ; Cytophagaceae/*physiology ; Female ; Japan ; Male ; Phylogeny ; Reproduction/physiology ; Sequence Analysis, DNA ; *Sex Ratio ; Symbiosis/genetics/*physiology ; Tetranychidae/*microbiology/physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Intracellular bacteria of the genus Wolbachia (alpha Proteobacteria) induce cytoplasmic incompatibility (CI) in many arthropod species, including spider mites, but not all Wolbachia cause CI. In spider mites CI becomes apparent by a reduced egg hatchability and a lower daughter:son ratio: CI in haplodiploid organisms in general was expected to produce all-male offspring or a male-biased sex ratio without any death of eggs. In a previous study of Japanese populations of Tetranychus urticae, two out of three green-form populations tested were infected with non-CI Wolbachia strains, whereas none of six red-form populations harbored Wolbachia. As the survey of Wolbachia infection in T. urticae is still fragmentary in Japan, we checked Wolbachia infection in thirty green-form populations and 29 red-form populations collected from a wide range of Japanese islands. For Wolbachia-infected populations, we tested the effects of Wolbachia on the reproductive traits and determined the phylogenetic relationships of the different strains of Wolbachia. All but one green-form populations were infected with Wolbachia and all strains belonged to the subgroup Ori when the wsp gene was used to determine the phylogenetic relationships of different strains of Wolbachia. Six out of 29 red-form populations harbored Wolbachia and the infected strains belonged to the subgroups Ori and Bugs. Twenty-four of 29 infected green-form populations and five of six infected red-form populations induced CI among the hosts. Thus, CI-Wolbachia strains are widespread in Japan, and no geographical trend was observed in the CI-Wolbachia. Although three red-form populations harbored other intracellular bacteria Cardinium, they did not affect host reproduction.}, } @article {pmid17439303, year = {2007}, author = {Weeks, AR and Turelli, M and Harcombe, WR and Reynolds, KT and Hoffmann, AA}, title = {From parasite to mutualist: rapid evolution of Wolbachia in natural populations of Drosophila.}, journal = {PLoS biology}, volume = {5}, number = {5}, pages = {e114}, pmid = {17439303}, issn = {1545-7885}, mesh = {Animals ; Base Sequence ; Biological Evolution ; California ; Cytoplasm ; Drosophila/*microbiology ; Female ; Fertility/physiology ; Infertility/microbiology ; Male ; Molecular Sequence Data ; Tetracycline/pharmacology ; Wolbachia/drug effects/*genetics ; }, abstract = {Wolbachia are maternally inherited bacteria that commonly spread through host populations by causing cytoplasmic incompatibility, often expressed as reduced egg hatch when uninfected females mate with infected males. Infected females are frequently less fecund as a consequence of Wolbachia infection. However, theory predicts that because of maternal transmission, these "parasites" will tend to evolve towards a more mutualistic association with their hosts. Drosophila simulans in California provided the classic case of a Wolbachia infection spreading in nature. Cytoplasmic incompatibility allowed the infection to spread through individual populations within a few years and from southern to northern California (more than 700 km) within a decade, despite reducing the fecundity of infected females by 15%-20% under laboratory conditions. Here we show that the Wolbachia in California D. simulans have changed over the last 20 y so that infected females now exhibit an average 10% fecundity advantage over uninfected females in the laboratory. Our data suggest smaller but qualitatively similar changes in relative fecundity in nature and demonstrate that fecundity-increasing Wolbachia variants are currently polymorphic in natural populations.}, } @article {pmid17371764, year = {2007}, author = {McGraw, EA and O'Neill, SL}, title = {Wolbachia: invasion biology in South pacific butterflies.}, journal = {Current biology : CB}, volume = {17}, number = {6}, pages = {R220-1}, doi = {10.1016/j.cub.2007.01.038}, pmid = {17371764}, issn = {0960-9822}, mesh = {Animals ; Biological Evolution ; Butterflies/genetics/*microbiology/physiology ; Embryo, Nonmammalian/microbiology ; Female ; Geography ; Male ; Pacific Ocean ; Sex Ratio ; Sexual Behavior, Animal ; Symbiosis ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachia ensdosymbionts are well known for their ability to manipulate the population biology and development of their hosts. One of the less studied outcomes of Wolbachia infection with this symbiont is the selective killing of male embryos. Recent work on butterflies living on different South Pacific islands is beginning to help us understand the complexity of the co-evolutionary interactions between these partners.}, } @article {pmid17364245, year = {2007}, author = {Pankewitz, F and Zöllmer, A and Hilker, M and Gräser, Y}, title = {Presence of Wolbachia in insect eggs containing antimicrobially active anthraquinones.}, journal = {Microbial ecology}, volume = {54}, number = {4}, pages = {713-721}, pmid = {17364245}, issn = {0095-3628}, mesh = {Animals ; Anthraquinones/analysis ; Anti-Bacterial Agents/analysis ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Coleoptera/chemistry/microbiology/physiology ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/analysis/isolation & purification ; *Hemiptera/chemistry/microbiology/physiology ; Molecular Sequence Data ; Ovum/chemistry/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Wolbachia are obligatory, cytoplasmatically inherited alpha-proteobacteria, which are common endosymbionts in arthropods where they may cause reproductive abnormalities. Many insects are well known to protect themselves from deleterious microorganisms by antibiotic components. In this study, we addressed the question whether Wolbachia are able to infect insects containing antimicrobial anthraquinones and anthrones, and if so, whether these genotypes of Wolbachia comprise a monophyletic cluster within one of the known supergroups. Leaf beetles of the taxon Galerucini (Galerucinae) are known to contain 1,8-dihydroxylated anthraquinones and anthrones. Also, the scale insect Dactylopius contains an anthraquinone glycoside, carminic acid. Our analyses revealed that a representative of the Galerucini, Galeruca tanaceti and Dactylopius, are indeed infected by endosymbiotic Wolbachia bacteria. Phylogenetic analysis of the wsp and ftsZ genes of these bacteria revealed that strains in G. tanaceti cluster in supergroup A, whereas those present in Dactylopius are distinctive from each other and from those of G. tanaceti. They are clustering in supergroups A and B. Wolbachia strains present in close, but anthraquinone-free relatives of G. tanaceti were shown to belong also to supergroup A. From these results, we can conclude (1) a double infection in Dactylopius, (2) that the presence of antimicrobial compounds such as anthraquinones does not necessarily protect insects from infection by Wolbachia, and (3) that genotypes of Wolbachia-infecting anthraquinone-containing insects most likely do not comprise a unique genotype. These results show that Wolbachia bacteria might be adapted to cope even with conditions usually detrimental to other bacteria and that these adaptations are widespread among Wolbachia supergroups.}, } @article {pmid17361572, year = {2004}, author = {Hernández, M and Quesada, T and Muñoz, C and Espinoza, AM}, title = {Genetic diversity of Costa Rican populations of the rice planthopper Tagosodes orizicolus (Homoptera: Delphacidae).}, journal = {Revista de biologia tropical}, volume = {52}, number = {3}, pages = {795-806}, pmid = {17361572}, issn = {0034-7744}, mesh = {Animals ; Costa Rica ; Female ; *Genetic Variation ; Hemiptera/classification/*genetics/microbiology ; Male ; Polymerase Chain Reaction ; Random Amplified Polymorphic DNA Technique ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Tagosodes orizicolus (Homoptera: Delphacidae) is one of the main constraints of the rice production in the Neotropics. This planthopper produces severe damages as a phloem feeder, causes mechanical injury during oviposition and vectors the rice hoja blanca virus (RHBV). The main objective of this study was to determine the genetic diversity of T. orizicolus populations from three rice growing regions of Costa Rica, using RAPDs. Individuals from Guanacaste, Parrita, San Carlos and Cali-Colombia, as outgroup, were analyzed using the random primers. Phenetic relationships revealed that the Costa Rican populations were clearly separated from Cali-Colombia, sharing less than 25% similarity. Costa Rican populations were divided into two main branches separated at 30% similarity. The first branch included Guanacaste and San Carlos and the second displayed Parrita. In relation to similarity indexes within groups, the Guanacaste cluster showed the highest (over 50%) and Cali-Colombia was the most diverse (28%). The correspondence analysis confirmed the clusters of the phenogram and showed close interactions between the Parrita and San Carlos populations. The genetic separation observed could be the result of the geographic isolation among populations, but it could also be explained by the infection with the rickettsia Wolbachia pipientis. This bacterium causes cytoplasmic incompatibility in its host, which results in non-viable progeny when infected males mate with non-infected females, or when insects hosting different strains of Wolbachia mate. Then, a search for Wolbachia in previously described populations of T orizicolus was initiated. The presence of the bacteria was analyzed by PCR with 16S rDNA-specific primers for Wolbachia. The PCR analyses revealed infections of 86% in the population of San Carlos, 96% in Guanacaste, 37% in Parrita and 100% in Cali-Colombia. Crosses between individuals of T. orizicolus from Parrita and Guanacaste were performed for testing cytoplasmic incompatibility. When infected males were crossed with non-infected females within the same population, a significant reduction in progeny number was obtained as well as when crosses between infected individuals belonging to different populations were performed. These experiments showed cytoplasmic incompatibility not only caused by the presence of Wolbachia within the population, but also by the presence of different strains of the bacteria between populations.}, } @article {pmid17350292, year = {2007}, author = {Vaishampayan, PA and Dhotre, DP and Gupta, RP and Lalwani, P and Ghate, H and Patole, MS and Shouche, YS}, title = {Molecular evidence and phylogenetic affiliations of Wolbachia in cockroaches.}, journal = {Molecular phylogenetics and evolution}, volume = {44}, number = {3}, pages = {1346-1351}, doi = {10.1016/j.ympev.2007.01.003}, pmid = {17350292}, issn = {1055-7903}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Bacteriophages/genetics/isolation & purification ; Base Sequence ; Cockroaches/genetics/*microbiology ; Cytoskeletal Proteins/genetics ; DNA Primers/genetics ; DNA, Bacterial/genetics ; Evolution, Molecular ; Genes, Bacterial ; Genes, Viral ; India ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Wolbachia/*classification/*genetics/isolation & purification/virology ; }, } @article {pmid17329802, year = {2007}, author = {Lo, N and Paraskevopoulos, C and Bourtzis, K and O'Neill, SL and Werren, JH and Bordenstein, SR and Bandi, C}, title = {Taxonomic status of the intracellular bacterium Wolbachia pipientis.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {57}, number = {Pt 3}, pages = {654-657}, doi = {10.1099/ijs.0.64515-0}, pmid = {17329802}, issn = {1466-5026}, mesh = {Animals ; Arthropods/microbiology/physiology ; Filarioidea/microbiology/physiology ; Wolbachia/*classification/genetics/physiology ; }, abstract = {Wolbachia pipientis is a maternally inherited, intracellular bacterium found in more than 20 % of all insects, as well as numerous other arthropods and filarial nematodes. It has been the subject of a growing number of studies in recent decades, because of the remarkable effects it has on its arthropod hosts, its potential as a tool for biological control of arthropods of agricultural and medical importance and its use as a target for treatment of filariasis. W. pipientis was originally discovered in cells of the mosquito Culex pipiens and is the only formally described member of the genus. Molecular sequence-based studies have revealed a number of phylogenetically diverse strains of W. pipientis. Owing to uncertainty about whether W. pipientis comprises more than one species, researchers in the field now commonly refer to W. pipientis simply as Wolbachia. In this note, we briefly review higher-level phylogenetic and recombination studies of W. pipientis and propose that all the intracellular symbionts known to cluster closely with the type strain of W. pipientis, including those in the currently recognized supergroups (A-H), are officially given this name.}, } @article {pmid17306254, year = {2007}, author = {Suba, N and Shiny, C and Taylor, MJ and Narayanan, RB}, title = {Brugia malayi Wolbachia hsp60 IgG antibody and isotype reactivity in different clinical groups infected or exposed to human bancroftian lymphatic filariasis.}, journal = {Experimental parasitology}, volume = {116}, number = {3}, pages = {291-295}, doi = {10.1016/j.exppara.2006.12.006}, pmid = {17306254}, issn = {0014-4894}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Adult ; Animals ; Antibodies, Bacterial/biosynthesis/blood ; Antibody Specificity ; Blotting, Western ; Brugia malayi/microbiology ; Chaperonin 60/genetics/*immunology ; Electrophoresis, Polyacrylamide Gel ; Elephantiasis, Filarial/*immunology/microbiology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin G/*biosynthesis/blood ; Male ; Recombinant Proteins/genetics/immunology ; Wolbachia/*immunology ; *Wuchereria bancrofti ; }, abstract = {Wolbachia, an endosymbiotic bacterium in filarial parasites, comes into contact with the host immune system upon parasite death. Here, we analyzed, total IgG and isotype antibody responses to Wolbachia hsp60 in individuals from an area endemic for Wuchereria bancrofti. Wolbachia derived hsp60 gene was cloned and the recombinant protein was used to determine the IgG and isotype reactivity by Western blotting and ELISA. All individuals from the endemic area generated antibody responses to Brugia malayi Wolbachia hsp60, which were elevated in the group with chronic pathology. Isotype analysis showed that, all clinical groups mounted IgG1-IgG4 responses with higher levels of B. malayi Wolbachia hsp60 specific IgG1 observed in the sera of patients with chronic pathology compared to microfilaraemics and endemic normals. These findings suggests that Wolbachia-derived hsp60 generates antibody responses in individuals infected or exposed to W. bancrofti and an elevated IgG and IgG1 reactivity is observed in people with filarial pathology.}, } @article {pmid17305835, year = {2007}, author = {Flor, M and Hammerstein, P and Telschow, A}, title = {Wolbachia-induced unidirectional cytoplasmic incompatibility and the stability of infection polymorphism in parapatric host populations.}, journal = {Journal of evolutionary biology}, volume = {20}, number = {2}, pages = {696-706}, doi = {10.1111/j.1420-9101.2006.01252.x}, pmid = {17305835}, issn = {1010-061X}, mesh = {Adaptation, Physiological ; Animal Migration ; Animals ; Female ; Fertility/physiology ; Infectious Disease Transmission, Vertical ; Male ; *Models, Biological ; Population Dynamics ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular, maternally inherited bacteria that are widespread among arthropods and commonly induce a reproductive incompatibility between infected male and uninfected female hosts known as unidirectional cytoplasmic incompatibility (CI). If infected and uninfected populations occur parapatrically, CI acts as a post-zygotic isolation barrier. We investigate the stability of such infection polymorphisms in a mathematical model with two populations linked by migration. We determine critical migration rates below which infected and uninfected populations can coexist. Analytical solutions of the critical migration rate are presented for mainland-island models. These serve as lower estimations for a more general model with two-way migration. The critical migration rate is positive if either Wolbachia causes a fecundity reduction in infected female hosts or its transmission is incomplete, and is highest for intermediate levels of CI. We discuss our results with respect to local adaptations of the Wolbachia host, speciation, and pest control.}, } @article {pmid17305834, year = {2007}, author = {Engelstädter, J and Hammerstein, P and Hurst, GD}, title = {The evolution of endosymbiont density in doubly infected host species.}, journal = {Journal of evolutionary biology}, volume = {20}, number = {2}, pages = {685-695}, doi = {10.1111/j.1420-9101.2006.01257.x}, pmid = {17305834}, issn = {1010-061X}, mesh = {Animals ; Bacterial Physiological Phenomena ; *Biological Evolution ; Female ; Fertility/physiology ; *Infectious Disease Transmission, Vertical ; Insecta/microbiology/physiology ; Male ; *Models, Biological ; Wolbachia/physiology ; }, abstract = {Multiple infection of individual hosts with several species or strains of maternally inherited endosymbionts is commonly observed in animals, especially insects. Here, we address theoretically the effect of co-infection on the optimal density of the endosymbionts in doubly infected hosts. Our analysis is based on the observation that a maternally inherited double infection is only stable if doubly infected females produce more doubly infected daughters than singly infected or uninfected females produce daughters. We consider both a general model and a model involving two endosymbionts inducing bidirectional cytoplasmic incompatibility (CI). We demonstrate that the optimal replication rate of endosymbionts in doubly infected hosts can be expected to be similar to or below the optimal replication rate in singly infected hosts. This is in contrast to some theoretical predictions for horizontally transmitted parasites and stems from the two strains of endosymbionts having coupled fitness. We discuss our results with respect to recent empirical results on endosymbiont densities, the evolution of CI-inducing bacteria and, more generally, the evolution of cooperation through direct fitness benefits.}, } @article {pmid17298560, year = {2007}, author = {Sanogo, YO and Dobson, SL and Bordenstein, SR and Novak, RJ}, title = {Disruption of the Wolbachia surface protein gene wspB by a transposable element in mosquitoes of the Culex pipiens complex (Diptera, Culicidae).}, journal = {Insect molecular biology}, volume = {16}, number = {2}, pages = {143-154}, doi = {10.1111/j.1365-2583.2006.00707.x}, pmid = {17298560}, issn = {0962-1075}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics/metabolism ; Base Sequence ; Culex/genetics/*microbiology ; *DNA Transposable Elements ; Female ; *Gene Silencing ; Geography ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Symbiosis/genetics ; United States ; Wolbachia/*genetics ; }, abstract = {Culex pipiens quinquefasciatus Say and Culex pipiens pipiens Linnaeus are sibling species incriminated as important vectors of emerging and re-emerging infectious diseases worldwide. The two forms differ little morphologically and are differentiated mainly based upon ecological, behavioural, physiological and genetic traits. Within the North American zone of sympatry, populations of Cx. p. quinquefasciatus and Cx. p. pipiens undergo extensive introgression and hybrid forms have been reported in nature. Both Cx. p. quinquefasciatus and Cx. p. pipiens are infected with the endosymbiotic bacteria Wolbachia pipientis. Here, we report the presence of a transposable element belonging to the IS256 family (IS256wPip) associated with Wolbachia in both Cx. p. quinquefasciatus and Cx. p. pipiens populations. Using reverse transcriptase PCR and sequence analysis, we show that IS256wPip has disrupted the wspB locus, a paralogue of the Wolbachia outer membrane protein (wspA) gene. The inactivation of the wspB appears to be specific to Cx. p. quinquefasciatus and to hybrids of the two forms, and was not observed in the surveyed Cx. p. pipiens mosquitoes. Our results support the hypothesis of a different origin of North American Cx. p. quinquefasciatus and Cx. p. pipiens populations. The flux of mobile genetic elements in the Wolbachia wPip genome could explain the high level of crossing types observed among different Culex populations. The insertion of IS256wPip into wspB may comprise a genetic candidate for discriminating Wolbachia symbionts in Culex.}, } @article {pmid17277413, year = {2007}, author = {Zhi, D and Keich, U and Pevzner, P and Heber, S and Tang, H}, title = {Correcting base-assignment errors in repeat regions of shotgun assembly.}, journal = {IEEE/ACM transactions on computational biology and bioinformatics}, volume = {4}, number = {1}, pages = {54-64}, doi = {10.1109/TCBB.2007.1005}, pmid = {17277413}, issn = {1545-5963}, support = {1 R01 HG02366-01/HG/NHGRI NIH HHS/United States ; }, mesh = {Algorithms ; Campylobacter jejuni/genetics ; Cluster Analysis ; Computational Biology/*methods ; Genome, Bacterial ; Lactococcus lactis/genetics ; *Models, Statistical ; Repetitive Sequences, Nucleic Acid/*genetics ; Sequence Alignment ; Sequence Analysis, DNA/*methods/statistics & numerical data ; Software ; Staphylococcus epidermidis/genetics ; Wolbachia/genetics ; }, abstract = {Accurate base-assignment in repeat regions of a whole genome shotgun assembly is an unsolved problem. Since reads in repeat regions cannot be easily attributed to a unique location in the genome, current assemblers may place these reads arbitrarily. As a result, the base-assignment error rate in repeats is likely to be much higher than that in the rest of the genome. We developed an iterative algorithm, EULER-AIR, that is able to correct base-assignment errors in finished genome sequences in public databases. The Wolbachia genome is among the best finished genomes. Using this genome project as an example, we demonstrated that EULER-AIR can 1) discover and correct base-assignment errors, 2) provide accurate read assignments, 3) utilize finishing reads for accurate base-assignment, and 4) provide guidance for designing finishing experiments. In the genome of Wolbachia, EULER-AIR found 16 positions with ambiguous base-assignment and two positions with erroneous bases. Besides Wolbachia, many other genome sequencing projects have significantly fewer finishing reads and, hence, are likely to contain more base-assignment errors in repeats. We demonstrate that EULER-AIR is a software tool that can be used to find and correct base-assignment errors in a genome assembly project.}, } @article {pmid17276921, year = {2007}, author = {Charlat, S and Reuter, M and Dyson, EA and Hornett, EA and Duplouy, A and Davies, N and Roderick, GK and Wedell, N and Hurst, GD}, title = {Male-killing bacteria trigger a cycle of increasing male fatigue and female promiscuity.}, journal = {Current biology : CB}, volume = {17}, number = {3}, pages = {273-277}, doi = {10.1016/j.cub.2006.11.068}, pmid = {17276921}, issn = {0960-9822}, mesh = {Animals ; Asia, Southeastern ; Australia ; Butterflies/genetics/*microbiology/*physiology ; Female ; Male ; Pacific Islands ; Sex Ratio ; *Sexual Behavior, Animal ; Wolbachia/*physiology ; }, abstract = {Sex-ratio distorters are found in numerous species and can reach high frequencies within populations. Here, we address the compelling, but poorly tested, hypothesis that the sex ratio bias caused by such elements profoundly alters their host's mating system. We compare aspects of female and male reproductive biology between island populations of the butterfly Hypolimnas bolina that show varying degrees of female bias, because of a male-killing Wolbachia infection. Contrary to expectation, female bias leads to an increase in female mating frequency, up to a point where male mating capacity becomes limiting. We show that increased female mating frequency can be explained as a facultative response to the depleted male mating resources in female biased populations. In other words, this system is one where male-killing bacteria trigger a vicious circle of increasing male fatigue and female promiscuity.}, } @article {pmid17276538, year = {2007}, author = {Jaenike, J}, title = {Fighting back against male-killers.}, journal = {Trends in ecology & evolution}, volume = {22}, number = {4}, pages = {167-169}, doi = {10.1016/j.tree.2007.01.008}, pmid = {17276538}, issn = {0169-5347}, mesh = {Animals ; Butterflies/*genetics/*microbiology ; Female ; Genetic Variation ; Inbreeding ; Male ; Sex Ratio ; Wolbachia/*pathogenicity/physiology ; }, abstract = {Male-killing endosymbionts create a genetic black hole into which host nuclear genes vanish. In a recent paper, Hornett et al. transferred male-killing Wolbachia between different strains of the butterfly Hypolimnas bolina through hybridization and backcrossing. Their results provide unambiguous evidence of genetic variation for resistance to male-killers. A possible consequence of such variation is that male-killing might appear and disappear quickly on an evolutionary timescale.}, } @article {pmid17264501, year = {2007}, author = {Pearlman, E and Gillette-Ferguson, I}, title = {Onchocerca volvulus, Wolbachia and river blindness.}, journal = {Chemical immunology and allergy}, volume = {92}, number = {}, pages = {254-265}, doi = {10.1159/000099276}, pmid = {17264501}, issn = {1660-2242}, support = {EY10320/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Humans ; Immunity, Innate ; Keratitis/immunology ; Onchocerca volvulus/*pathogenicity ; Onchocerciasis, Ocular/*etiology/immunology/pathology ; *Symbiosis ; Toll-Like Receptors/physiology ; Wolbachia/*pathogenicity ; }, abstract = {Chronic infection with filarial nematodes results in development of a suppressive response to an immense parasite burden, thereby limiting pathological and clinical manifestations. However, pro-inflammatory responses to dead and degenerating Onchocerca volvulus worms and release of endosymbiotic Wolbachia bacteria result in corneal opacification, sacrification and visual impairment. This review discusses host and parasite factors implicated in maintaining this balance of pro- an anti-inflammatory responses, and will focus on adaptive and innate immunity to filarial antigens and endosymbiotic Wolbachia bacteria.}, } @article {pmid17255232, year = {2007}, author = {Nfon, CK and Makepeace, BL and Njongmeta, LM and Tanya, VN and Trees, AJ}, title = {Lack of resistance after re-exposure of cattle cured of Onchocerca ochengi infection with oxytetracycline.}, journal = {The American journal of tropical medicine and hygiene}, volume = {76}, number = {1}, pages = {67-72}, pmid = {17255232}, issn = {0002-9637}, mesh = {Animals ; Anthelmintics/*therapeutic use ; Arsenicals/therapeutic use ; Cattle ; Cattle Diseases/*drug therapy ; Disease Susceptibility/*veterinary ; Female ; Onchocerciasis/drug therapy/*veterinary ; Oxytetracycline/*therapeutic use ; Time Factors ; Triazines/therapeutic use ; }, abstract = {Although vector control and ivermectin chemotherapy have led to a dramatic reduction in the incidence of river blindness (onchocerciasis), there is a consensus that additional control tools are required to sustain and extend this success. The recognition of endosymbiotic bacteria (Wolbachia) in filariae and their targeting by antibiotics constitutes the most significant and practicable opportunity for a macrofilaricidal therapy in the short-to-medium-term. Using Onchocerca ochengi in cattle, an analog of human onchocerciasis, we have previously shown that oxytetracycline is macrofilaricidal, and protective immunity exists naturally in a subset of animals termed putatively immune. Here, we report that although 24 weeks of weekly oxytetracycline treatment eliminated adult worms, cured animals remained susceptible to re-infection by natural challenge when compared with putatively immune cattle. However, their susceptibility was not significantly different from that of concurrently exposed, heavily infected animals. Thus, cattle cured by oxytetracycline are neither hypo-susceptible nor hyper-susceptible.}, } @article {pmid17254505, year = {2007}, author = {Johnston, KL and Taylor, MJ}, title = {Wolbachia in filarial parasites: targets for filarial infection and disease control.}, journal = {Current infectious disease reports}, volume = {9}, number = {1}, pages = {55-59}, pmid = {17254505}, issn = {1523-3847}, abstract = {Lymphatic filariasis and onchocerciasis are debilitating diseases caused by parasitic filarial nematodes. These nematodes have evolved a mutualistic symbiosis with intracellular bacteria of the genus Wolbachia, which are required for nematode embryogenesis and survival. The essential role of these bacteria in the biology of the nematode and their demonstrated involvement in the pathogenesis of filariasis make Wolbachia a promising novel chemotherapeutic target for the control of filarial infection and disease. This article reviews the recent findings, which highlight potential processes that form the basis of the symbiosis, the role of Wolbachia in filarial pathogenesis, and the efficacy of Wolbachia-targeted antibiotic chemotherapy in human trials. Future prospects for the development of an anti-Wolbachia treatment regimen suitable for integration into mass drug administration programs are also discussed.}, } @article {pmid17251127, year = {2007}, author = {Charlat, S and Davies, N and Roderick, GK and Hurst, GD}, title = {Disrupting the timing of Wolbachia-induced male-killing.}, journal = {Biology letters}, volume = {3}, number = {2}, pages = {154-156}, pmid = {17251127}, issn = {1744-9561}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Embryo, Nonmammalian/microbiology ; Female ; Male ; Moths/*microbiology ; Rifampin/pharmacology ; Sex Factors ; Sex Ratio ; Tetracycline/pharmacology ; Time Factors ; Wolbachia/drug effects/*pathogenicity ; }, abstract = {Several lineages of maternally inherited symbionts have evolved the ability to kill infected females' sons, a phenomenon known as male-killing. Male-killing varies in its timing, from early (death during embryogenesis) to late (mortality of late larval instars). Following the observation that treatment of male-killer infected adult females Hypolimnas bolina with tetracycline, a bacteriostatic antibiotic, produces a delay in the timing of male death, we hypothesized that early male-killers possess the ability to kill males through bacterial activity outside of embryogenesis. We verified this hypothesis by showing that treatment of surviving larvae with the bacteriocidal antibiotic rifampicin rescues males. This discounted the hypothesis that delayed death occurred due to postponed effects of toxins produced at earlier stages, and thus supported the importance of bacterial activity in the larval phase in delayed male-killing. These results argue against the view that early male-killing is achieved by specifically targeting an early developmental process within the sex determination pathway.}, } @article {pmid17251124, year = {2007}, author = {Mouton, L and Henri, H and Charif, D and Boulétreau, M and Vavre, F}, title = {Interaction between host genotype and environmental conditions affects bacterial density in Wolbachia symbiosis.}, journal = {Biology letters}, volume = {3}, number = {2}, pages = {210-213}, pmid = {17251124}, issn = {1744-9561}, mesh = {Analysis of Variance ; Animals ; Breeding ; Female ; Genotype ; Host-Pathogen Interactions/physiology ; Linear Models ; Male ; Population Density ; Symbiosis/*physiology ; *Temperature ; Wasps/*genetics/*microbiology ; Wolbachia/*physiology ; }, abstract = {Regulation of microbial population density is a necessity in stable symbiotic interactions. In Wolbachia symbiosis, both bacterial and host genotypes are involved in density regulation, but environmental factors may also affect bacterial population density. Here, we studied the interaction between three strains of Wolbachia in two divergent homozygous lines of the wasp Leptopilina heterotoma at two different temperatures. Wolbachia density varied between the two host genotypes at only one temperature. Moreover, at this temperature, reciprocal-cross F1 insects displayed identical Wolbachia densities, which were intermediate between the densities in the two parental lines. While these findings confirm that the host genotype plays an important role in Wolbachia density, they also highlight its interaction with environmental conditions, making possible the evolution of local adaptations for the regulation of Wolbachia density.}, } @article {pmid17241395, year = {2007}, author = {Bazzocchi, C and Comazzi, S and Santoni, R and Bandi, C and Genchi, C and Mortarino, M}, title = {Wolbachia surface protein (WSP) inhibits apoptosis in human neutrophils.}, journal = {Parasite immunology}, volume = {29}, number = {2}, pages = {73-79}, doi = {10.1111/j.1365-3024.2006.00915.x}, pmid = {17241395}, issn = {0141-9838}, mesh = {Animals ; Annexin A5 ; Apoptosis/*drug effects ; Bacterial Outer Membrane Proteins/*pharmacology ; Caspase 3/metabolism ; Fluorescein-5-isothiocyanate ; Humans ; In Situ Nick-End Labeling ; Interleukin-8/metabolism ; Nematoda/microbiology ; Neutrophils/*drug effects/physiology ; Wolbachia/metabolism ; }, abstract = {Polymorphonuclear cells (PMNs) are essential for the innate immune response against invading bacteria. At the same time, modulation of PMNs' apoptosis or cell death by bacteria has emerged as a mechanism of pathogenesis. Wolbachia bacteria are Gram-negative endosymbionts of filarial nematodes and arthropods, phylogenetically related to the genera Anaplasma, Ehrlichia and Neorickettsia (family Anaplasmataceae). Although several pathogens are known to interfere with apoptosis, there is only limited information on specific proteins that modulate this phenomenon. This is the first evidence for the anti-apoptotic activity of a surface protein of Wolbachia from filarial nematode parasites (the Wolbachia surface protein, WSP). The inhibition of apoptosis was demonstrated on purified human PMNs in vitro by different methods. TUNEL assay showed that the percentage of dead cells was reduced after stimulation with WSP; Annexin V-FITC binding assay confirmed that cell death was due mainly to apoptosis and not to necrosis. Reduced caspase-3 activity in stimulated cells also confirmed an inhibition of the apoptotic process.}, } @article {pmid17226040, year = {2007}, author = {Rymaszewska, A}, title = {Symbiotic bacteria in oocyte and ovarian cell mitochondria of the tick Ixodes ricinus: biology and phylogenetic position.}, journal = {Parasitology research}, volume = {100}, number = {5}, pages = {917-920}, pmid = {17226040}, issn = {0932-0113}, mesh = {Animals ; Bacteria/*classification/genetics/isolation & purification ; Bacterial Physiological Phenomena ; Ixodes/*microbiology/physiology ; Mitochondria/*microbiology ; Oocytes/*microbiology ; Phylogeny ; Symbiosis ; }, abstract = {Under natural conditions, eukaryote cells may contain bacteria. Arthropods such as ticks, insects or mites are a group particularly favoured by the obligate intracellular bacteria. While arthropods are vectors for some of them, other bacteria inhabit invertebrate host cells having entered mutualistic interactions. Such endosymbionts dwell usually in the host cell vacuoles or cytoplasm but have been also reported from tick oocyte mitochondria. The microorganisms contribute to mitochondria degradation, but their colonies are not eliminated from the tick cells affected. So far, such bacteria have been detected in three research centres. The Italian centre has reported on results of microscope and molecular analyses, while the Polish centre published molecular data. The Danish centre registered a 16S rRNA gene fragment in GenBank. Independent comparisons of the 16S rRNA gene sequences, carried out in the Italian and Polish centres, confirmed that the nucleotide sequences of the Ixodes ricinus endosymbionts formed a single clade with certain non-identified tick bacterium species isolated from the tick Haemaphysalis wellingtoni. On the other hand, pathogenic species of the genera Anaplasma, Ehrlichia and Rickettsia detected in I. ricinus as well as symbionts of the genus Wolbachia present in Culex pipiens and Drosophila simulans have been placed at a different site on the phylogenetic tree.}, } @article {pmid17216550, year = {2007}, author = {Narita, S and Nomura, M and Kato, Y and Yata, O and Kageyama, D}, title = {Molecular phylogeography of two sibling species of Eurema butterflies.}, journal = {Genetica}, volume = {131}, number = {3}, pages = {241-253}, doi = {10.1007/s10709-006-9134-1}, pmid = {17216550}, issn = {0016-6707}, mesh = {Animals ; Bacterial Infections/genetics/microbiology ; Butterflies/*genetics ; DNA, Mitochondrial/genetics ; Electron Transport Complex IV/genetics ; Evolution, Molecular ; Asia, Eastern ; Genetic Predisposition to Disease ; Genetic Variation ; *Geography ; NADH Dehydrogenase/genetics ; *Phylogeny ; Wolbachia ; }, abstract = {The common yellow butterfly Eurema hecabe is widely distributed in East Asia, and is one of the most burdensome species for taxonomists due to the numerous geographic and seasonal wing colour patterns. Moreover, within this species, individuals with a yellow wing fringe that occur in temperate regions of Japan (Y type) proved to be biologically different from others that occur widely in subtropical regions of Japan and all over East Asia (B type). To unveil the genetic variation within and between the two types, a total of 50 butterflies collected at 18 geographic localities in East Asia were examined for nucleotide sequence variation of three mitochondrial regions: cytochrome c oxidase subunit I (COI), cytochrome c oxidase subunit III (COIII) and NADH dehydrogenase subunit 5 (ND5). In addition, they were also examined for infection status with the endosymbiotic bacteria Wolbachia. The three mitochondrial sequences consistently showed that (i) Y type and B type were highly divergent, (ii) nucleotide variation within B type was very small although sampled from a geographically wide range, and (iii) a weak association existed between mitochondrial DNA haplotypes and Wolbachia infection status.}, } @article {pmid17216316, year = {2007}, author = {Simón, F and Kramer, LH and Román, A and Blasini, W and Morchón, R and Marcos-Atxutegi, C and Grandi, G and Genchi, C}, title = {Immunopathology of Dirofilaria immitis infection.}, journal = {Veterinary research communications}, volume = {31}, number = {2}, pages = {161-171}, pmid = {17216316}, issn = {0165-7380}, mesh = {Animals ; Cat Diseases/immunology/microbiology/*parasitology/pathology ; Cats ; Dirofilaria immitis/*immunology ; Dirofilariasis/*immunology/pathology ; Dog Diseases/immunology/microbiology/*parasitology/pathology ; Dogs ; Humans ; Immunity, Innate/immunology ; Th1 Cells/immunology/microbiology/parasitology ; Th2 Cells/immunology/microbiology/parasitology ; Wolbachia/immunology ; }, abstract = {Heartworm disease caused by Dirofilaria immitis affects canine and feline hosts, with infections occasionally being reported in humans. Studies have shown that both dirofilarial antigens and those derived from its bacterial endosymbiont Wolbachia, interact with the host organism during canine, feline and human infections and participate in the development of the pathology and in the regulation of the host's immune response. Both innate and acquired immune responses are observed and the development of the acquired response may depend on the host and, or on its parasitological status. This review aims at illustrating current research on the role of both D. immitis and Wolbachia, in the immunology and immunopathology of dirofilariosis.}, } @article {pmid17202370, year = {2007}, author = {Hise, AG and Daehnel, K and Gillette-Ferguson, I and Cho, E and McGarry, HF and Taylor, MJ and Golenbock, DT and Fitzgerald, KA and Kazura, JW and Pearlman, E}, title = {Innate immune responses to endosymbiotic Wolbachia bacteria in Brugia malayi and Onchocerca volvulus are dependent on TLR2, TLR6, MyD88, and Mal, but not TLR4, TRIF, or TRAM.}, journal = {Journal of immunology (Baltimore, Md. : 1950)}, volume = {178}, number = {2}, pages = {1068-1076}, doi = {10.4049/jimmunol.178.2.1068}, pmid = {17202370}, issn = {0022-1767}, support = {AI 07024/AI/NIAID NIH HHS/United States ; EY 10320/EY/NEI NIH HHS/United States ; EY 11373/EY/NEI NIH HHS/United States ; K08 AI 054652/AI/NIAID NIH HHS/United States ; }, mesh = {Adaptor Proteins, Vesicular Transport/immunology ; Animals ; Brugia malayi/*immunology ; Cell Line ; Humans ; Immunity, Innate/*immunology ; Inflammation/immunology/microbiology ; Interleukin-6/biosynthesis ; Macrophages/immunology/metabolism ; Membrane Transport Proteins/*immunology ; Mice ; Mice, Inbred C57BL ; Myelin Proteins/*immunology ; Myelin and Lymphocyte-Associated Proteolipid Proteins ; Myeloid Differentiation Factor 88/*immunology ; Onchocerca volvulus/*immunology ; Proteolipids/*immunology ; Receptors, Interleukin/immunology ; Rickettsiaceae Infections/immunology/microbiology ; Symbiosis/immunology ; Toll-Like Receptor 2/immunology ; Toll-Like Receptor 4/immunology ; Toll-Like Receptor 6/immunology ; Toll-Like Receptors/*immunology ; Tumor Necrosis Factor-alpha/biosynthesis ; Wolbachia/*immunology/isolation & purification ; Wuchereria/immunology ; }, abstract = {The discovery that endosymbiotic Wolbachia bacteria play an important role in the pathophysiology of diseases caused by filarial nematodes, including lymphatic filariasis and onchocerciasis (river blindness) has transformed our approach to these disabling diseases. Because these parasites infect hundreds of millions of individuals worldwide, understanding host factors involved in the pathogenesis of filarial-induced diseases is paramount. However, the role of early innate responses to filarial and Wolbachia ligands in the development of filarial diseases has not been fully elucidated. To determine the role of TLRs, we used cell lines transfected with human TLRs and macrophages from TLR and adaptor molecule-deficient mice and evaluated macrophage recruitment in vivo. Extracts of Brugia malayi and Onchocerca volvulus, which contain Wolbachia, directly stimulated human embryonic kidney cells expressing TLR2, but not TLR3 or TLR4. Wolbachia containing filarial extracts stimulated cytokine production in macrophages from C57BL/6 and TLR4(-/-) mice, but not from TLR2(-/-) or TLR6(-/-) mice. Similarly, macrophages from mice deficient in adaptor molecules Toll/IL-1R domain-containing adaptor-inducing IFN-beta and Toll/IL-1R domain-containing adaptor-inducing IFN-beta-related adaptor molecule produced equivalent cytokines as wild-type cells, whereas responses were absent in macrophages from MyD88(-/-) and Toll/IL-1R domain-containing adaptor protein (TIRAP)/MyD88 adaptor-like (Mal) deficient mice. Isolated Wolbachia bacteria demonstrated similar TLR and adaptor molecule requirements. In vivo, macrophage migration to the cornea in response to filarial extracts containing Wolbachia was dependent on TLR2 but not TLR4. These results establish that the innate inflammatory pathways activated by endosymbiotic Wolbachia in B. malayi and O. volvulus filaria are dependent on TLR2-TLR6 interactions and are mediated by adaptor molecules MyD88 and TIRAP/Mal.}, } @article {pmid17190825, year = {2007}, author = {Pannebakker, BA and Loppin, B and Elemans, CP and Humblot, L and Vavre, F}, title = {Parasitic inhibition of cell death facilitates symbiosis.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {104}, number = {1}, pages = {213-215}, pmid = {17190825}, issn = {0027-8424}, mesh = {Animals ; *Apoptosis ; Female ; *Oogenesis ; *Symbiosis ; Wasps/*parasitology ; Wolbachia/*physiology ; }, abstract = {Symbiotic microorganisms have had a large impact on eukaryotic evolution, with effects ranging from parasitic to mutualistic. Mitochondria and chloroplasts are prime examples of symbiotic microorganisms that have become obligate for their hosts, allowing for a dramatic extension of suitable habitats for life. Out of the extraordinary diversity of bacterial endosymbionts in insects, most are facultative for their hosts, such as the ubiquitous Wolbachia, which manipulates host reproduction. Some endosymbionts, however, have become obligatory for host reproduction and/or survival. In the parasitoid wasp Asobara tabida the presence of Wolbachia is necessary for host oogenesis, but the mechanism involved is yet unknown. We show that Wolbachia influences programmed cell death processes (a host regulatory feature typically targeted by pathogens) in A. tabida, making its presence essential for the wasps' oocytes to mature. This suggests that parasite strategies, such as bacterial regulation of host apoptosis, can drive the evolution of host dependence, allowing for a swift transition from parasitism to mutualism.}, } @article {pmid17174921, year = {2006}, author = {Charlat, S and Engelstädter, J and Dyson, EA and Hornett, EA and Duplouy, A and Tortosa, P and Davies, N and Roderick, GK and Wedell, N and Hurst, GD}, title = {Competing selfish genetic elements in the butterfly Hypolimnas bolina.}, journal = {Current biology : CB}, volume = {16}, number = {24}, pages = {2453-2458}, doi = {10.1016/j.cub.2006.10.062}, pmid = {17174921}, issn = {0960-9822}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Asia, Southeastern ; Butterflies/*genetics/*microbiology/physiology ; Cytoplasm/*genetics ; DNA, Bacterial ; Female ; Geography ; Male ; Molecular Sequence Data ; Phenotype ; *Repetitive Sequences, Nucleic Acid ; Reproduction/genetics/physiology ; Species Specificity ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/drug effects/*genetics/physiology ; }, abstract = {Maternally inherited selfish genetic elements are common in animals . Whereas host genetics and ecology are recognized as factors that may limit the incidence of these parasites , theory suggests one further factor-interference with other selfish elements-that could affect their prevalence . In this paper, we show that spatial heterogeneity in the occurrence of the male-killing Wolbachia wBol1 in the tropical butterfly Hypolimnas bolina is caused by a second infection that can exclude the male-killer. We first provide evidence of a second Wolbachia strain, wBol2, present in most populations that do not carry the male-killer but rare or absent when the male-killer is present. Crossing data indicate that wBol2 in males induces cytoplasmic incompatibility to both uninfected and wBol1-infected females. The wBol2 infection can therefore not only spread through uninfected populations but also resist invasion by wBol1. Thus, we provide empirical support for the hypothesis that the incidence of particular selfish genetic elements can limit the presence of competing types.}, } @article {pmid17174110, year = {2007}, author = {Lo, N and Evans, TA}, title = {Phylogenetic diversity of the intracellular symbiont Wolbachia in termites.}, journal = {Molecular phylogenetics and evolution}, volume = {44}, number = {1}, pages = {461-466}, doi = {10.1016/j.ympev.2006.10.028}, pmid = {17174110}, issn = {1055-7903}, mesh = {Animals ; *Genetic Variation ; Isoptera/*microbiology ; Molecular Sequence Data ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Symbiosis/genetics ; Wolbachia/classification/*genetics/growth & development ; }, } @article {pmid17151264, year = {2007}, author = {Dowling, DK and Friberg, U and Hailer, F and Arnqvist, G}, title = {Intergenomic epistasis for fitness: within-population interactions between cytoplasmic and nuclear genes in Drosophila melanogaster.}, journal = {Genetics}, volume = {175}, number = {1}, pages = {235-244}, pmid = {17151264}, issn = {0016-6731}, mesh = {Animals ; Cell Nucleus/*metabolism ; Chromosomes/genetics ; Cytoplasm/*metabolism ; DNA, Mitochondrial/*genetics/metabolism ; Drosophila Proteins/*genetics ; Drosophila melanogaster/*genetics/metabolism ; *Epistasis, Genetic ; Female ; Genotype ; Male ; Polymorphism, Genetic ; Selection, Genetic ; Sex Distribution ; }, abstract = {The symbiotic relationship between the mitochondrial and nuclear genomes coordinates metabolic energy production and is fundamental to life among eukaryotes. Consequently, there is potential for strong selection to shape interactions between these two genomes. Substantial research attention has focused on the possibility that within-population sequence polymorphism in mitochondrial DNA (mtDNA) is maintained by mitonuclear fitness interactions. Early theory predicted that selection will often eliminate mitochondrial polymorphisms. However, recent models demonstrate that intergenomic interactions can promote the maintenance of polymorphism, especially if the nuclear genes involved are linked to the X chromosome. Most empirical studies to date that have assessed cytonuclear fitness interactions have studied variation across populations and it is still unclear how general and strong such interactions are within populations. We experimentally tested for cytonuclear interactions within a laboratory population of Drosophila melanogaster using 25 randomly sampled cytoplasmic genomes, expressed in three different haploid nuclear genetic backgrounds, while eliminating confounding effects of intracellular bacteria (e.g., Wolbachia). We found sizable cytonuclear fitness interactions within this population and present limited evidence suggesting that these effects were sex specific. Moreover, the relative fitness of cytonuclear genotypes was environment specific. Sequencing of mtDNA (2752 bp) revealed polymorphism within the population, suggesting that the observed cytoplasmic genetic effects may be mitochondrial in origin.}, } @article {pmid17151259, year = {2007}, author = {Engelstädter, J and Hurst, GD}, title = {The impact of male-killing bacteria on host evolutionary processes.}, journal = {Genetics}, volume = {175}, number = {1}, pages = {245-254}, pmid = {17151259}, issn = {0016-6731}, mesh = {Animals ; *Biological Evolution ; Computer Simulation ; Female ; Gene Flow ; Insecta/growth & development/*microbiology ; Male ; Models, Biological ; Polymorphism, Genetic ; *Selection, Genetic ; Sex Ratio ; *Symbiosis ; Wolbachia/genetics/growth & development/*pathogenicity ; }, abstract = {Male-killing bacteria are maternally inherited endosymbionts that selectively kill male offspring of their arthropod hosts. Using both analytical techniques and computer simulations, we studied the impact of these bacteria on the population genetics of their hosts. In particular, we derived and corroborated formulas for the fixation probability of mutant alleles, mean times to fixation and fixation or extinction, and heterozygosity for varying male-killer prevalence. Our results demonstrate that infections with male-killing bacteria impede the spread of beneficial alleles, facilitate the spread of deleterious alleles, and reduce genetic variation. The reason for this lies in the strongly reduced fitness of infected females combined with no or very limited gene flow from infected females to uninfected individuals. These two properties of male-killer-infected populations reduce the population size relevant for the initial emergence and spread of mutations. In contrast, use of Wright's equation relating sex ratio to effective population size produces misleading predictions. We discuss the relationship to the similar effect of background selection, the impact of other sex-ratio-distorting endosymbionts, and how our results affect the interpretation of empirical data on genetic variation in male-killer-infected populations.}, } @article {pmid17148240, year = {2005}, author = {Kondo, N and Shimada, M and Fukatsu, T}, title = {Infection density of Wolbachia endosymbiont affected by co-infection and host genotype.}, journal = {Biology letters}, volume = {1}, number = {4}, pages = {488-491}, pmid = {17148240}, issn = {1744-9561}, mesh = {Animals ; Coleoptera/genetics/*microbiology ; Genotype ; Polymerase Chain Reaction ; Symbiosis/*physiology ; Wolbachia/growth & development/*physiology ; }, abstract = {Infection density is among the most important factors for understanding the biological effects of Wolbachia and other endosymbionts on their hosts. To gain insight into the mechanisms of infection density regulation, we investigated the adzuki bean beetles Callosobruchus chinensis and their Wolbachia endosymbionts. Double-infected, single-infected and uninfected host strains with controlled nuclear genetic backgrounds were generated by introgression, and infection densities in these strains were evaluated by a quantitative polymerase chain reaction technique. Our study revealed previously unknown aspects of Wolbachia density regulation: (i) the identification of intra-specific host genotypes that affect Wolbachia density differently and (ii) the suppression of Wolbachia density by co-infecting Wolbachia strains. These findings shed new light on symbiont-symbiont and host-symbiont interactions in the Wolbachia-insect endosymbiosis and strongly suggest that Wolbachia density is determined through a complex interaction between host genotype, symbiont genotype and other factors.}, } @article {pmid17126589, year = {2007}, author = {Morchón, R and Bazzocchi, C and López-Belmonte, J and Martín-Pacho, JR and Kramer, LH and Grandi, G and Simón, F}, title = {iNOs expression is stimulated by the major surface protein (rWSP) from Wolbachia bacterial endosymbiont of Dirofilaria immitis following subcutaneous injection in mice.}, journal = {Parasitology international}, volume = {56}, number = {1}, pages = {71-75}, doi = {10.1016/j.parint.2006.10.003}, pmid = {17126589}, issn = {1383-5769}, mesh = {Animals ; Antibodies, Bacterial/biosynthesis ; Bacterial Outer Membrane Proteins/*immunology ; Chaperonin 60/immunology ; Dirofilaria immitis/*microbiology/physiology ; Dirofilariasis/immunology/parasitology ; Inflammation/immunology ; Interferon-gamma/genetics/immunology ; Interleukin-10/genetics ; Interleukin-4/genetics ; Mice ; Mice, Inbred BALB C ; Nitric Oxide/metabolism ; Nitric Oxide Synthase Type II/genetics/*metabolism ; RNA, Messenger ; Recombinant Proteins/immunology ; Symbiosis ; Th1 Cells/immunology ; Wolbachia/*immunology/physiology ; }, abstract = {The bacterial endosymbiont Wolbachia of several species of filarial nematodes plays an important role in the inflammatory pathology of filariasis. Nitric oxide (NO) production has also been implicated in the immune response during filarial infections. Here we present data indicating that a recombinant Wolbachia surface protein (rWSP) induces iNOs mRNA expression and NO production, as well as IFN-gamma and a Th1-type antibody response, in inoculated BALB/c mice. This effect is not observed when mice are inoculated with a recombinant heat shock protein from Wolbachia (GroEL).}, } @article {pmid17119612, year = {2005}, author = {Puttaraju, HP and Prakash, BM}, title = {Effects of Wolbachia in the uzifly, Exorista sorbillans, a parasitoid of the silkworm, Bombyx mori.}, journal = {Journal of insect science (Online)}, volume = {5}, number = {}, pages = {30}, pmid = {17119612}, issn = {1536-2442}, mesh = {Animals ; Bombyx/*parasitology ; Diptera/*microbiology ; Dose-Response Relationship, Drug ; Female ; Male ; Oxytetracycline/pharmacology ; Reproduction ; Sex Ratio ; Wolbachia/drug effects/*physiology ; }, abstract = {The uzifly, Exorista sorbillans (Diptera: Tachinidae), a parasitoid of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), harbours Wolbachia (Rickettsia) endosymbionts. Administration of 0.05 mg/ml oxytetracycline to the adult uziflies removed Wolbachia endosymbionts and resulted in different reproductive disorders, such as i) reduction in fecundity of uninfected females, ii) cytoplasmic incompatibility in crosses between infected males and uninfected females, iii) sterility in the crosses between both males and females from uninfected populations, and iv) sex-ratio distortion in uninfected females irrespective of the presence of Wolbachia in males. However, tetracycline treatment did not have much effect on longevity of the uzifly. These results suggest that the interaction of Wolbachia with its uzifly host is one of mutual symbiosis as it controls the reproductive physiology of its hosts.}, } @article {pmid17113827, year = {2007}, author = {Pfarr, KM and Hoerauf, A}, title = {A niche for Wolbachia.}, journal = {Trends in parasitology}, volume = {23}, number = {1}, pages = {5-7}, doi = {10.1016/j.pt.2006.11.002}, pmid = {17113827}, issn = {1471-4922}, mesh = {Animals ; Arthropods/*microbiology ; Drosophila/microbiology ; Female ; Nematoda/*microbiology ; Phenotype ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbionts of arthropods and filarial nematodes. Arthropods infected with these endobacteria display altered reproductive phenotypes, including cytoplasmic incompatibility and sex-ratio distortion. In nematodes, the endobacteria are essential for embryogenesis and worm survival. Wolbachia are transmitted vertically from mother to progeny, and Frydman et al. recently showed that, after transfer to uninfected Drosophila, Wolbachia rapidly accumulate in the somatic stem cell niche. From this location, the endobacteria might enter the developing oocytes and infect the progeny.}, } @article {pmid17112544, year = {2007}, author = {Vautrin, E and Charles, S and Genieys, S and Vavre, F}, title = {Evolution and invasion dynamics of multiple infections with Wolbachia investigated using matrix based models.}, journal = {Journal of theoretical biology}, volume = {245}, number = {2}, pages = {197-209}, doi = {10.1016/j.jtbi.2006.09.035}, pmid = {17112544}, issn = {0022-5193}, mesh = {Animals ; *Biological Evolution ; Female ; Genetic Variation ; Gram-Negative Bacterial Infections/microbiology/*transmission/*veterinary ; Infectious Disease Transmission, Vertical ; Insecta/*microbiology ; Male ; Models, Biological ; Sex Factors ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Endosymbiotic bacteria are often transmitted vertically from one host generation to the next via oocytes cytoplasm. The generally small number of colonizing bacteria in the oocytes leads to a bottleneck at each generation, resulting in genetic homogenization of the symbiotic population. Nevertheless, in many of the species infected by Wolbachia (maternally transmitted bacteria), individuals do sometimes simultaneously harbor several bacterial strains, owing to the fact that Wolbachia induces cytoplasmic incompatibility (CI) that maintains multiple infections. CI occurs in crosses in which the male is infected by at least one Wolbachia strain that the female lacks, and consequently it favors individuals with the greatest symbiotic diversity. CI results in death of offspring in diploid species. In haplodiploid individuals, unfertilized eggs hatch normally into males and fertilized ones, which would lead to females, either die (female mortality type: FM) or develop into males (male development type: MD). Until now, only one theoretical study, restricted to diploid species, has investigated the associations where multiple CI-inducing Wolbachia co-exist, and explored the conditions under which multiple infections can spread. The consequences of double infections on Wolbachia maintenance in host populations, and the selective pressures to which it is subjected have not yet been analysed. Here, we have re-written a model previously developed for single infection in matrix form, which allows easy extension to multiple infections and introduction of mutant strains. We show that (i) the CI type has a strong influence on invasiveness and maintenance of multiple infections; (ii) double infection lowers the invasion threshold of less competitive strains that hitch-hike with their companion strain; (iii) when multiple infections occur, as in single infections, the strains selected are those which maximize the production of infected offspring; and (iv) for the MD CI type, invasion of mutant strains can carry the whole infection to extinction.}, } @article {pmid17095536, year = {2007}, author = {Gavotte, L and Henri, H and Stouthamer, R and Charif, D and Charlat, S and Boulétreau, M and Vavre, F}, title = {A Survey of the bacteriophage WO in the endosymbiotic bacteria Wolbachia.}, journal = {Molecular biology and evolution}, volume = {24}, number = {2}, pages = {427-435}, doi = {10.1093/molbev/msl171}, pmid = {17095536}, issn = {0737-4038}, mesh = {Animals ; Bacteriophage Typing ; Bacteriophages/classification/*genetics ; Capsid Proteins/genetics ; DNA, Viral ; *Evolution, Molecular ; Gene Transfer, Horizontal ; Genes, Viral ; Insecta/microbiology ; Molecular Sequence Data ; Nematoda/microbiology ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/classification/genetics/*virology ; }, abstract = {Bacteriophages are common viruses infecting prokaryotes. In addition to their deadly effect, phages are also involved in several evolutionary processes of bacteria, such as coding functional proteins potentially beneficial to them, or favoring horizontal gene transfer through transduction. The particular lifestyle of obligatory intracellular bacteria usually protects them from phage infection. However, Wolbachia, an intracellular alpha-proteobacterium, infecting diverse arthropod and nematode species and best known for the reproductive alterations it induces, harbors a phage named WO, which has recently been proven to be lytic. Here, phage infection was checked in 31 Wolbachia strains, which induce 5 different effects in their hosts and infect 25 insect species and 3 nematodes. Only the Wolbachia infecting nematodes and Trichogramma were found devoid of phage infection. All the 25 detected phages were characterized by the DNA sequence of a minor capsid protein gene. Based on all data currently available, phylogenetic analyses show a lack of congruency between Wolbachia or insect and phage WO phylogenies, indicating numerous horizontal transfers of phage among the different Wolbachia strains. The absence of relation between phage phylogeny and the effects induced by Wolbachia suggests that WO is not directly involved in these effects. Implications on phage WO evolution are discussed.}, } @article {pmid17054511, year = {2006}, author = {Takiya, DM and Tran, PL and Dietrich, CH and Moran, NA}, title = {Co-cladogenesis spanning three phyla: leafhoppers (Insecta: Hemiptera: Cicadellidae) and their dual bacterial symbionts.}, journal = {Molecular ecology}, volume = {15}, number = {13}, pages = {4175-4191}, doi = {10.1111/j.1365-294X.2006.03071.x}, pmid = {17054511}, issn = {0962-1083}, mesh = {Animals ; DNA, Ribosomal ; Evolution, Molecular ; Flavobacteriaceae/physiology ; Gammaproteobacteria/physiology ; *Genetic Speciation ; Hemiptera/*genetics/*microbiology ; Host-Parasite Interactions ; *Phylogeny ; *Symbiosis ; Trees/physiology ; Wolbachia/physiology ; }, abstract = {Endosymbioses are a major form of biological complexity affecting the ecological and evolutionary diversification of many eukaryotic groups. These associations are exemplified by nutritional symbioses of insects for which phylogenetic studies have demonstrated numerous cases of long-term codiversification between a bacterial and a host lineage. Some insects, including most leafhoppers (Insecta: Hemiptera: Cicadellidae), have more than one bacterial symbiont within specialized host cells, raising questions regarding the patterns of codiversification of these multiple partners and the evolutionary persistence of complex symbiotic systems. Previous studies reported the presence of two dominant symbiont types in a member of the leafhopper subfamily Cicadellinae (sharpshooters). In this study, 16S rRNA sequences were obtained and used to examine the occurrence and evolutionary relationships of the two dominant symbiont types across 29 leafhopper species. Candidatus Sulcia muelleri (Bacteroidetes) was detected in all leafhopper species examined, a finding that is consistent with a previous report of its ancient association with the Auchenorrhyncha (a grouping that includes leafhoppers, treehoppers, cicadas, planthoppers, and spittlebugs). Baumannia cicadellinicola (Proteobacteria), previously known from only five sharpshooter species, was found only in the sharpshooter tribes Cicadellini and Proconiini, as well as in the subfamily Phereurhininae. Mitochondrial and nuclear gene sequences were obtained and used to reconstruct host phylogenies. Analyses of host and symbiont data sets support a congruent evolutionary history between sharpshooters, Sulcia and Baumannia and thus provide the first strong evidence for long-term co-inheritance of multiple symbionts during the diversification of a eukaryotic host. Sulcia shows a fivefold lower rate of 16S rDNA sequence divergence than does Baumannia for the same host pairs. The term 'coprimary' symbiont is proposed for such cases.}, } @article {pmid17048002, year = {2007}, author = {Covacin, C and Barker, SC}, title = {Supergroup F Wolbachia bacteria parasitise lice (Insecta: Phthiraptera).}, journal = {Parasitology research}, volume = {100}, number = {3}, pages = {479-485}, pmid = {17048002}, issn = {0932-0113}, mesh = {Animals ; Base Sequence ; Molecular Sequence Data ; Phthiraptera/*microbiology ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal/genetics ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {We studied six species of lice from three of the four suborders of lice. These lice were infected with Wolbachia bacteria from supergroups A and F. This is the first report of an infection of supergroup F Wolbachia in lice. To date, Wolbachia from supergroup F have been found in filarial nematodes, Mansonella spp., and, rarely, in insects. We inferred the phylogeny of the Wolbachia from lice and representatives of all Wolbachia supergroups, with nucleotide sequences from the small subunit ribosomal RNA gene (SSU rRNA). There was no evidence of congruence between the taxon of louse and the Wolbachia bacteria that infect lice. There is no evidence that Wolbachia and their louse hosts co-evolved at least at the level of Wolbachia supergroups. We propose a novel mechanism for the horizontal transfer of Wolbachia between different species of lice from birds: transfer of Wolbachia during phoresis by hippoboscid flies.}, } @article {pmid17044881, year = {2006}, author = {Wiwatanaratanabutr, S and Kittayapong, P}, title = {Effects of temephos and temperature on Wolbachia load and life history traits of Aedes albopictus.}, journal = {Medical and veterinary entomology}, volume = {20}, number = {3}, pages = {300-307}, doi = {10.1111/j.1365-2915.2006.00640.x}, pmid = {17044881}, issn = {0269-283X}, mesh = {Aedes/*drug effects/*microbiology ; Animals ; Dose-Response Relationship, Drug ; *Hot Temperature ; Insecticides/*pharmacology ; Larva/drug effects/growth & development ; Life Cycle Stages ; Temefos/*pharmacology ; Wings, Animal/growth & development ; Wolbachia/drug effects/*isolation & purification ; }, abstract = {Maternally inherited Wolbachia (gram-negative bacteria) often affect the reproductive fitness of their arthropod hosts and may cause cytoplasmic incompatibility (CI). Comparing Wolbachia-infected and uninfected strains of the mosquito Aedes albopictus (Skuse) (Diptera: Culicidae), we assessed the effects on fitness of two stressors: temperature elevation (25 degrees C vs. 37 degrees C) and exposure to temephos insecticide (concentration range 0.0017-0.0167 mg/L) during larval development. Fitness was measured in terms of life history traits: percentage survival, development time and wing size. Insecticide treatment was associated with reduction in survival rates and wing size in both sexes, but did not affect development time or Wolbachia load. Temperature elevation by 12 degrees C significantly reduced all four bionomic parameters observed in both sexes. Wolbachia density within individual adult mosquitoes was determined by using real-time quantitative polymerase chain reaction (PCR) based on the wsp gene. Both male and female adults had significantly lower densities of Wolbachia after larval rearing at the higher temperature.}, } @article {pmid17044733, year = {2006}, author = {Debrah, AY and Mand, S and Specht, S and Marfo-Debrekyei, Y and Batsa, L and Pfarr, K and Larbi, J and Lawson, B and Taylor, M and Adjei, O and Hoerauf, A}, title = {Doxycycline reduces plasma VEGF-C/sVEGFR-3 and improves pathology in lymphatic filariasis.}, journal = {PLoS pathogens}, volume = {2}, number = {9}, pages = {e92}, pmid = {17044733}, issn = {1553-7374}, mesh = {Adult ; Animals ; Doxycycline/*therapeutic use ; Elephantiasis, Filarial/blood/*drug therapy/parasitology/*pathology ; Female ; Filaricides/*therapeutic use ; Humans ; Intercellular Signaling Peptides and Proteins/blood ; Lymphatic System/diagnostic imaging/drug effects ; Male ; Microfilariae/isolation & purification/physiology ; Middle Aged ; Molecular Sequence Data ; Parasitemia/parasitology ; Testicular Hydrocele/parasitology ; Testis/diagnostic imaging ; Treatment Outcome ; Ultrasonography ; Vascular Endothelial Growth Factor C/antagonists & inhibitors/*blood ; Vascular Endothelial Growth Factor Receptor-3/antagonists & inhibitors/*blood ; Wolbachia/isolation & purification ; }, abstract = {Lymphatic filariasis is a disease of considerable socioeconomic burden in the tropics. Presently used antifilarial drugs are able to strongly reduce transmission and will thus ultimately lower the burden of morbidity associated with the infection, however, a chemotherapeutic principle that directly induces a halt or improvement in the progression of the morbidity in already infected individuals would constitute a major lead. In search of such a more-effective drug to complement the existing ones, in an area endemic for bancroftian filariasis in Ghana, 33 microfilaremic and 18 lymphedema patients took part in a double-blind, placebo-controlled trial of a 6-wk regimen of 200 mg/day doxycycline. Four months after doxycycline treatment, all patients received 150-200 microg/kg ivermectin and 400 mg albendazole. Patients were monitored for Wolbachia and microfilaria loads, antigenemia, filarial dance sign (FDS), dilation of supratesticular lymphatic vessels, and plasma levels of lymphangiogenic factors (vascular endothelial growth factor-C [VEGF-C] and soluble vascular endothelial growth factor receptor-3 [(s)VEGFR-3]). Lymphedema patients were additionally monitored for stage (grade) of lymphedema and the circumferences of affected legs. Wolbachia load, microfilaremia, antigenemia, and frequency of FDS were significantly reduced in microfilaremic patients up to 24 mo in the doxycycline group compared to the placebo group. The mean dilation of supratesticular lymphatic vessels in doxycycline-treated patients was reduced significantly at 24 mo, whereas there was no improvement in the placebo group. Preceding clinical improvement, at 12 mo, the mean plasma levels of VEGF-C and sVEGFR-3 decreased significantly in the doxycycline-treated patients to a level close to that of endemic normal values, whereas there was no significant reduction in the placebo patients. The extent of disease in lymphedema patients significantly improved following doxycycline, with the mean stage of lymphedema in the doxycycline-treated patients being significantly lower compared to placebo patients 12 mo after treatment. The reduction in the stages manifested as better skin texture, a reduction of deep folds, and fewer deep skin folds. In conclusion, a 6-wk regimen of antifilarial treatment with doxycycline against W. bancrofti showed a strong macrofilaricidal activity and reduction in plasma levels of VEGF-C/sVEGFR-3, the latter being associated with amelioration of supratesticular dilated lymphatic vessels and with an improvement of pathology in lymphatic filariasis patients.}, } @article {pmid17040562, year = {2006}, author = {Haine, ER and Martin, J and Cook, JM}, title = {Deep mtDNA divergences indicate cryptic species in a fig-pollinating wasp.}, journal = {BMC evolutionary biology}, volume = {6}, number = {}, pages = {83}, pmid = {17040562}, issn = {1471-2148}, mesh = {Animals ; Cytochromes b/genetics ; DNA, Mitochondrial/*genetics ; Ficus/*physiology ; *Genetic Speciation ; Genetic Variation/*genetics ; Male ; Molecular Sequence Data ; Phylogeny ; Pollen/*physiology ; Symbiosis ; Wasps/*classification/cytology/genetics/*physiology ; }, abstract = {BACKGROUND: Figs and fig-pollinating wasps are obligate mutualists that have coevolved for ca. 90 million years. They have radiated together, but do not show strict cospeciation. In particular, it is now clear that many fig species host two wasp species, so there is more wasp speciation than fig speciation. However, little is known about how fig wasps speciate.

RESULTS: We studied variation in 71 fig-pollinating wasps from across the large geographic range of Ficus rubiginosa in Australia. All wasps sampled belong to one morphological species (Pleistodontes imperialis), but we found four deep mtDNA clades that differed from each other by 9-17% nucleotides. As these genetic distances exceed those normally found within species and overlap those (10-26%) found between morphologically distinct Pleistodontes species, they strongly suggest cryptic fig wasp species. mtDNA clade diversity declines from all four present in Northern Queensland to just one in Sydney, near the southern range limit. However, at most sites multiple clades coexist and can be found in the same tree or even the same fig fruit and there is no evidence for parallel sub-division of the host fig species. Both mtDNA data and sequences from two nuclear genes support the monophyly of the "P. imperialis complex" relative to other Pleistodontes species, suggesting that fig wasp divergence has occurred without any host plant shift. Wasps in clade 3 were infected by a single strain (W1) of Wolbachia bacteria, while those in other clades carried a double infection (W2+W3) of two other strains.

CONCLUSION: Our study indicates that cryptic fig-pollinating wasp species have developed on a single host plant species, without the involvement of host plant shifts, or parallel host plant divergence. Despite extensive evidence for coevolution between figs and fig wasps, wasp speciation may not always be linked strongly with fig speciation.}, } @article {pmid17040394, year = {2006}, author = {de Crespigny, FE and Pitt, TD and Wedell, N}, title = {Increased male mating rate in Drosophila is associated with Wolbachia infection.}, journal = {Journal of evolutionary biology}, volume = {19}, number = {6}, pages = {1964-1972}, doi = {10.1111/j.1420-9101.2006.01143.x}, pmid = {17040394}, issn = {1010-061X}, mesh = {Animals ; Drosophila/*microbiology/physiology ; Drosophila melanogaster/microbiology/physiology ; Female ; Male ; Rickettsiaceae Infections/physiopathology ; *Sexual Behavior, Animal ; Wolbachia/*physiology ; }, abstract = {The maternally inherited bacterium Wolbachia pipientis infects 25-75% of arthropods and manipulates host reproduction to improve its transmission. One way Wolbachia achieves this is by inducing cytoplasmic incompatibility (CI), where crosses between infected males and uninfected females are inviable. Infected males suffer reduced fertility through CI and reduced sperm production. However, Wolbachia induce lower levels of CI in nonvirgin males. We examined the impact of Wolbachia on mating behaviour in male Drosophila melanogaster and D. simulans, which display varying levels of CI, and show that infected males mate at a higher rate than uninfected males in both species. This may serve to increase the spread of Wolbachia, or alternatively, may be a behavioural adaptation employed by males to reduce the level of CI. Mating at high rate restores reproductive compatibility with uninfected females resulting in higher male reproductive success thus promoting male promiscuity. Increased male mating rates also have implications for the transmission of Wolbachia.}, } @article {pmid17040125, year = {2006}, author = {Fenn, K and Conlon, C and Jones, M and Quail, MA and Holroyd, NE and Parkhill, J and Blaxter, M}, title = {Phylogenetic relationships of the Wolbachia of nematodes and arthropods.}, journal = {PLoS pathogens}, volume = {2}, number = {10}, pages = {e94}, pmid = {17040125}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Arthropods/*microbiology ; Base Sequence ; DNA, Bacterial ; Evolution, Molecular ; Gene Transfer, Horizontal ; *Genome, Bacterial ; Molecular Sequence Data ; Nematoda/*microbiology ; Phylogeny ; Symbiosis/genetics ; Synteny ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia are well known as bacterial symbionts of arthropods, where they are reproductive parasites, but have also been described from nematode hosts, where the symbiotic interaction has features of mutualism. The majority of arthropod Wolbachia belong to clades A and B, while nematode Wolbachia mostly belong to clades C and D, but these relationships have been based on analysis of a small number of genes. To investigate the evolution and relationships of Wolbachia symbionts we have sequenced over 70 kb of the genome of wOvo, a Wolbachia from the human-parasitic nematode Onchocerca volvulus, and compared the genes identified to orthologues in other sequenced Wolbachia genomes. In comparisons of conserved local synteny, we find that wBm, from the nematode Brugia malayi, and wMel, from Drosophila melanogaster, are more similar to each other than either is to wOvo. Phylogenetic analysis of the protein-coding and ribosomal RNA genes on the sequenced fragments supports reciprocal monophyly of nematode and arthropod Wolbachia. The nematode Wolbachia did not arise from within the A clade of arthropod Wolbachia, and the root of the Wolbachia clade lies between the nematode and arthropod symbionts. Using the wOvo sequence, we identified a lateral transfer event whereby segments of the Wolbachia genome were inserted into the Onchocerca nuclear genome. This event predated the separation of the human parasite O. volvulus from its cattle-parasitic sister species, O. ochengi. The long association between filarial nematodes and Wolbachia symbionts may permit more frequent genetic exchange between their genomes.}, } @article {pmid17036209, year = {2006}, author = {Paraskevopoulos, C and Bordenstein, SR and Wernegreen, JJ and Werren, JH and Bourtzis, K}, title = {Toward a Wolbachia multilocus sequence typing system: discrimination of Wolbachia strains present in Drosophila species.}, journal = {Current microbiology}, volume = {53}, number = {5}, pages = {388-395}, pmid = {17036209}, issn = {0343-8651}, support = {R01 GM62626-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Typing Techniques/*methods ; Drosophila/*microbiology ; Genetic Variation ; Recombination, Genetic ; *Sequence Analysis, DNA ; Wolbachia/*classification ; }, abstract = {Among the diverse maternally inherited symbionts in arthropods, Wolbachia are the most common and infect over 20% of all species. In a departure from traditional genotyping or phylogenetic methods relying on single Wolbachia genes, the present study represents an initial Multilocus Sequence Typing (MLST) analysis to discriminate closely related Wolbachia pipientis strains, and additional data on sequence diversity in Wolbachia. We report a new phylogenetic characterization of four genes (aspC, atpD, sucB, and pdhB), and provide an expanded analysis of markers described in previous studies (16S rDNA, ftsZ, groEL, dnaA, and gltA). MLST analysis of the bacterial strains present in 16 different Drosophila-Wolbachia associations detected four distinct clonal complexes that also corresponded to maximum-likelihood identified phylogenetic clades. Among the 16 associations analyzed, six could not be assigned to MLST clonal complexes and were also shown to be in conflict with relationships predicted by maximum-likelihood phylogenetic inferences. The results demonstrate the discriminatory power of MLST for identifying strains and clonal lineages of Wolbachia and provide a robust foundation for studying the ecology and evolution of this widespread endosymbiont.}, } @article {pmid17035954, year = {2007}, author = {Gotoh, T and Noda, H and Ito, S}, title = {Cardinium symbionts cause cytoplasmic incompatibility in spider mites.}, journal = {Heredity}, volume = {98}, number = {1}, pages = {13-20}, doi = {10.1038/sj.hdy.6800881}, pmid = {17035954}, issn = {0018-067X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Bacteroidetes/drug effects/*genetics ; Cluster Analysis ; Crosses, Genetic ; Cytoplasm/*microbiology/physiology ; DNA, Ribosomal/genetics ; Female ; Male ; Phylogeny ; Reproduction/*physiology ; Sex Ratio ; *Symbiosis ; Tetranychidae/*microbiology/*physiology ; }, abstract = {Intracellular symbiotic bacteria belonging to the Cytophaga-Flavobacterium-Bacteroides lineage have recently been described and are widely distributed in arthropod species. The newly discovered bacteria, named Cardinium sp, cause the expression of various reproductive alterations in their arthropod hosts, including cytoplasmic incompatibility (CI), induction of parthenogenesis and feminization of genetic males. We detected 16S ribosomal DNA sequences similar to those of Cardinium from seven populations of five spider mite species, suggesting a broad distribution of infection of Cardinium in spider mites. To clarify the effect of Cardinium on the reproductive traits of the infected spider mites, infected mites were crossed with uninfected mites for each population. In one of the populations, Eotetranychus suginamensis, CI was induced when infected males were crossed with uninfected females. The other six populations of four species showed no reproductive abnormalities in the F(1) generation, but the possibility of CI effects in the F(2) generation remains to be tested. One species of spider mite, Tetranychus pueraricola, harbored both Cardinium and Wolbachia, but these symbionts seemed to have no effect on the reproduction of the host, even when the host was infected independently with each symbiont.}, } @article {pmid17032063, year = {2006}, author = {Jaenike, J and Dyer, KA and Cornish, C and Minhas, MS}, title = {Asymmetrical reinforcement and Wolbachia infection in Drosophila.}, journal = {PLoS biology}, volume = {4}, number = {10}, pages = {e325}, pmid = {17032063}, issn = {1545-7885}, mesh = {Animals ; *Biological Evolution ; Crosses, Genetic ; DNA, Mitochondrial/metabolism ; Drosophila/*genetics/*microbiology ; Female ; Genetic Variation ; *Hybridization, Genetic ; Male ; Molecular Sequence Data ; Reproduction ; Species Specificity ; Wolbachia/*pathogenicity ; }, abstract = {Reinforcement refers to the evolution of increased mating discrimination against heterospecific individuals in zones of geographic overlap and can be considered a final stage in the speciation process. One the factors that may affect reinforcement is the degree to which hybrid matings result in the permanent loss of genes from a species' gene pool. Matings between females of Drosophila subquinaria and males of D. recens result in high levels of offspring mortality, due to interspecific cytoplasmic incompatibility caused by Wolbachia infection of D. recens. Such hybrid inviability is not manifested in matings between D. recens females and D. subquinaria males. Here we ask whether the asymmetrical hybrid inviability is associated with a corresponding asymmetry in the level of reinforcement. The geographic ranges of D. recens and D. subquinaria were found to overlap across a broad belt of boreal forest in central Canada. Females of D. subquinaria from the zone of sympatry exhibit much stronger levels of discrimination against males of D. recens than do females from allopatric populations. In contrast, such reproductive character displacement is not evident in D. recens, consistent with the expected effects of unidirectional cytoplasmic incompatibility. Furthermore, there is substantial behavioral isolation within D. subquinaria, because females from populations sympatric with D. recens discriminate against allopatric conspecific males, whereas females from populations allopatric with D. recens show no discrimination against any conspecific males. Patterns of general genetic differentiation among populations are not consistent with patterns of behavioral discrimination, which suggests that the behavioral isolation within D. subquinaria results from selection against mating with Wolbachia-infected D. recens. Interspecific cytoplasmic incompatibility may contribute not only to post-mating isolation, an effect already widely recognized, but also to reinforcement, particularly in the uninfected species. The resulting reproductive character displacement not only increases behavioral isolation from the Wolbachia-infected species, but may also lead to behavioral isolation between populations of the uninfected species. Given the widespread occurrence of Wolbachia among insects, it thus appears that there are multiple ways by which these endosymbionts may directly and indirectly contribute to reproductive isolation and speciation.}, } @article {pmid17028229, year = {2006}, author = {Rasgon, JL and Ren, X and Petridis, M}, title = {Can Anopheles gambiae be infected with Wolbachia pipientis? Insights from an in vitro system.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {12}, pages = {7718-7722}, pmid = {17028229}, issn = {0099-2240}, mesh = {Animals ; Anopheles/*microbiology ; Cell Line ; Immunocompetence ; In Situ Hybridization, Fluorescence ; Insect Vectors/microbiology ; Malaria/prevention & control/transmission ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/isolation & purification/*pathogenicity ; }, abstract = {Wolbachia pipientis are maternally inherited endosymbionts associated with cytoplasmic incompatibility, a potential mechanism to drive transgenic traits into Anopheles populations for malaria control. W. pipientis infections are common in many mosquito genera but have never been observed in any Anopheles species, leading to the hypothesis that Anopheles mosquitoes are incapable of harboring infection. We used an in vitro system to evaluate the ability of Anopheles gambiae cells to harbor diverse W. pipientis infections. We successfully established W. pipientis infections (strains wRi and wAlbB) in the immunocompetent Anopheles gambiae cell line Sua5B. Infection was confirmed by PCR, antibiotic curing, DNA sequencing, and direct observation using fluorescence in situ hybridization. The infections were maintained at high passage rates for >30 passages. Our results indicate that there is no intrinsic genetic block to W. pipientis infection in A. gambiae cells, suggesting that establishment of in vivo W. pipientis infections in Anopheles mosquitoes may be feasible.}, } @article {pmid17028222, year = {2006}, author = {Morchón, R and López-Belmonte, J and Rodríguez-Barbero, A and Simón, F}, title = {High levels of serum thromboxane B2 are generated during human pulmonary dirofilariosis.}, journal = {Clinical and vaccine immunology : CVI}, volume = {13}, number = {10}, pages = {1175-1176}, pmid = {17028222}, issn = {1556-6811}, mesh = {Animals ; Dirofilaria immitis ; Dirofilariasis/*blood ; Dogs ; Humans ; Lung Diseases, Parasitic/*blood ; Thromboxane B2/biosynthesis/*blood ; }, abstract = {The canine parasite Dirofilaria immitis can infect humans. Patients with pulmonary dirofilariosis develop significantly higher thromboxane B2 levels than healthy individuals living in areas where dirofilariosis is endemic and in areas where dirofilariosis is not endemic. The possible role of Wolbachia bacteria in the appearance of this eicosanoid is discussed.}, } @article {pmid16962357, year = {2006}, author = {Nfon, CK and Makepeace, BL and Njongmeta, LM and Tanya, VN and Bain, O and Trees, AJ}, title = {Eosinophils contribute to killing of adult Onchocerca ochengi within onchocercomata following elimination of Wolbachia.}, journal = {Microbes and infection}, volume = {8}, number = {12-13}, pages = {2698-2705}, doi = {10.1016/j.micinf.2006.07.017}, pmid = {16962357}, issn = {1286-4579}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; Cattle ; Cell Degranulation ; Eosinophils/*immunology ; Histocytochemistry ; Leukocyte Count ; Onchocerca/*immunology/*microbiology ; Onchocerciasis/drug therapy/*immunology/pathology ; Oxytetracycline/*pharmacology/therapeutic use ; Symbiosis ; Wolbachia/*drug effects ; }, abstract = {Many filarial nematodes, including Onchocerca volvulus (the cause of human 'River Blindness'), have a mutually dependent relationship with Wolbachia bacteria. There has been much interest in Wolbachia as a chemotherapeutic target, since there are no macrofilaricidal drugs (i.e., lethal to adult worms) of low toxicity. Using the bovine parasite O. ochengi, we previously demonstrated that combined intensive and intermittent (COM) oxytetracycline treatment induces a sustained depletion of Wolbachia and is macrofilaricidal, whereas a short intensive regimen (SIR) is non-macrofilaricidal. To understand how targeting Wolbachia with oxytetracycline can lead to worm death, O. ochengi nodules (onchocercomata) were sequentially excised from cattle administered COM or SIR therapy, and cell infiltrates were microscopically quantified. Pre-treatment, worms were surrounded by neutrophils, with eosinophils rare or absent. At 8-12weeks after either regimen, eosinophils increased around worms and were observed degranulating on the cuticle. However, with the SIR treatment, neutrophils returned to predominance by 48weeks, while in the COM group, eosinophilia persisted. These observations suggest that accumulation of degranulating eosinophils over a prolonged period is a cause rather than an effect of parasite death, and the macrofilaricidal mechanism of antibiotics may relate to facilitation of eosinophil infiltration around worms by ablation of Wolbachia-mediated neutrophilia.}, } @article {pmid16950907, year = {2006}, author = {Weiss, BL and Mouchotte, R and Rio, RV and Wu, YN and Wu, Z and Heddi, A and Aksoy, S}, title = {Interspecific transfer of bacterial endosymbionts between tsetse fly species: infection establishment and effect on host fitness.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {11}, pages = {7013-7021}, pmid = {16950907}, issn = {0099-2240}, support = {T32AI07404/AI/NIAID NIH HHS/United States ; F32AI062680/AI/NIAID NIH HHS/United States ; AI51584/AI/NIAID NIH HHS/United States ; F32 AI062680/AI/NIAID NIH HHS/United States ; GM069449/GM/NIGMS NIH HHS/United States ; R01 AI051584/AI/NIAID NIH HHS/United States ; T32 AI007404/AI/NIAID NIH HHS/United States ; T01CT122306/CT/CIT NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; Enterobacteriaceae/genetics/*growth & development/*pathogenicity ; Female ; Insect Vectors/classification/growth & development/microbiology ; Phylogeny ; Polymerase Chain Reaction ; Species Specificity ; *Symbiosis ; Trypanosomiasis/prevention & control/transmission ; Tsetse Flies/classification/*growth & development/*microbiology ; }, abstract = {Tsetse flies (Glossina spp.) can harbor up to three distinct species of endosymbiotic bacteria that exhibit unique modes of transmission and evolutionary histories with their host. Two mutualist enterics, Wigglesworthia and Sodalis, are transmitted maternally to tsetse flies' intrauterine larvae. The third symbiont, from the genus Wolbachia, parasitizes developing oocytes. In this study, we determined that Sodalis isolates from several tsetse fly species are virtually identical based on a phylogenetic analysis of their ftsZ gene sequences. Furthermore, restriction fragment-length polymorphism analysis revealed little variation in the genomes of Sodalis isolates from tsetse fly species within different subgenera (Glossina fuscipes fuscipes and Glossina morsitans morsitans). We also examined the impact on host fitness of transinfecting G. fuscipes fuscipes and G. morsitans morsitans flies with reciprocal Sodalis strains. Tsetse flies cleared of their native Sodalis symbionts were successfully repopulated with the Sodalis species isolated from a different tsetse fly species. These transinfected flies effectively transmitted the novel symbionts to their offspring and experienced no detrimental fitness effects compared to their wild-type counterparts, as measured by longevity and fecundity. Quantitative PCR analysis revealed that transinfected flies maintained their Sodalis populations at densities comparable to those in flies harboring native symbionts. Our ability to transinfect tsetse flies is indicative of Sodalis ' recent evolutionary history with its tsetse fly host and demonstrates that this procedure may be used as a means of streamlining future paratransgenesis experiments.}, } @article {pmid16950898, year = {2006}, author = {Rasgon, JL and Gamston, CE and Ren, X}, title = {Survival of Wolbachia pipientis in cell-free medium.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {11}, pages = {6934-6937}, pmid = {16950898}, issn = {0099-2240}, mesh = {Aedes/cytology/microbiology ; Animals ; Bacteriological Techniques ; Cell Line ; *Culture Media ; DNA, Bacterial ; Host-Parasite Interactions ; In Situ Hybridization, Fluorescence ; Polymerase Chain Reaction ; Wolbachia/genetics/*growth & development ; }, abstract = {Wolbachia pipientis is an obligate intracellular bacterium found in a wide range of invertebrate taxa. While over ecological timescales Wolbachia infections are maintained by strict maternal inheritance, horizontal transfer events are common over evolutionary time. To be horizontally transferred between organisms, Wolbachia bacteria must pass through and survive an extracellular phase. We used BacLight live-dead staining, PCR, and fluorescence in situ hybridization to assess the ability for purified Wolbachia bacteria to survive in cell-free media. We found that purified Wolbachia bacteria were able to survive extracellularly for up to 1 week with no decrease in viability. While no replication was observed in the extracellular phase, purified Wolbachia bacteria were able to reinvade cells and establish stable infections at all time points. The ability of Wolbachia bacteria to survive outside host cells may increase the probability of successful horizontal transfer and the exploitation of new ecological niches. Our development of methods to purify and maintain viable Wolbachia bacteria from cultured cells will be useful for other researchers studying Wolbachia biology.}, } @article {pmid16936055, year = {2006}, author = {Baldo, L and Dunning Hotopp, JC and Jolley, KA and Bordenstein, SR and Biber, SA and Choudhury, RR and Hayashi, C and Maiden, MC and Tettelin, H and Werren, JH}, title = {Multilocus sequence typing system for the endosymbiont Wolbachia pipientis.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {11}, pages = {7098-7110}, pmid = {16936055}, issn = {0099-2240}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Alleles ; Animals ; Arthropods/classification/*microbiology ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/*genetics ; *Bacterial Typing Techniques ; DNA, Bacterial/analysis/isolation & purification ; Genetic Variation ; Molecular Sequence Data ; Recombination, Genetic ; *Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Wolbachia/*classification/genetics/growth & development ; }, abstract = {The eubacterial genus Wolbachia comprises one of the most abundant groups of obligate intracellular bacteria, and it has a host range that spans the phyla Arthropoda and Nematoda. Here we developed a multilocus sequence typing (MLST) scheme as a universal genotyping tool for Wolbachia. Internal fragments of five ubiquitous genes (gatB, coxA, hcpA, fbpA, and ftsZ) were chosen, and primers that amplified across the major Wolbachia supergroups found in arthropods, as well as other divergent lineages, were designed. A supplemental typing system using the hypervariable regions of the Wolbachia surface protein (WSP) was also developed. Thirty-seven strains belonging to supergroups A, B, D, and F obtained from singly infected hosts were characterized by using MLST and WSP. The number of alleles per MLST locus ranged from 25 to 31, and the average levels of genetic diversity among alleles were 6.5% to 9.2%. A total of 35 unique allelic profiles were found. The results confirmed that there is a high level of recombination in chromosomal genes. MLST was shown to be effective for detecting diversity among strains within a single host species, as well as for identifying closely related strains found in different arthropod hosts. Identical or similar allelic profiles were obtained for strains harbored by different insect species and causing distinct reproductive phenotypes. Strains with similar WSP sequences can have very different MLST allelic profiles and vice versa, indicating the importance of the MLST approach for strain identification. The MLST system provides a universal and unambiguous tool for strain typing, population genetics, and molecular evolutionary studies. The central database for storing and organizing Wolbachia bacterial and host information can be accessed at http://pubmlst.org/wolbachia/.}, } @article {pmid16933972, year = {2006}, author = {Hornett, EA and Charlat, S and Duplouy, AM and Davies, N and Roderick, GK and Wedell, N and Hurst, GD}, title = {Evolution of male-killer suppression in a natural population.}, journal = {PLoS biology}, volume = {4}, number = {9}, pages = {e283}, pmid = {16933972}, issn = {1545-7885}, mesh = {Animals ; *Biological Evolution ; Butterflies/genetics/*microbiology ; Female ; Genetic Variation ; Male ; Ovum ; *Sex Ratio ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Male-killing bacteria are widespread in arthropods, and can profoundly alter the reproductive biology of their host species. Here we detail the first case of complete suppression of a male killer. The nymphalid butterfly Hypolimnas bolina is infected with a strain of the bacterium Wolbachia, wBol1, which kills male host embryos in Polynesian populations, but does not do so in many areas of Southeast Asia, where both males and female adults are naturally infected, and wBol1-infected females produce a 1:1 sex ratio. We demonstrate that absence of male killing by wBol1 is associated with dominant zygotic suppression of the action of the male killer. Simulations demonstrate host suppressors of male-killer action can spread very rapidly, and historical data indicating the presence of male killing in Southeast Asia in the very recent past suggests suppressor spread has been a very recent occurrence. Thus, male killer/host interactions are much more dynamic than previously recognised, with rapid and dramatic loss of the phenotype. Our results also indicate that suppression can render male killers completely quiescent, leading to the conclusion that some species that do not currently express a male killer may have done so in the past, and thus that more species have had their biology affected by these parasites than previously believed.}, } @article {pmid16928646, year = {2006}, author = {Negri, I and Pellecchia, M and Mazzoglio, PJ and Patetta, A and Alma, A}, title = {Feminizing Wolbachia in Zyginidia pullula (Insecta, Hemiptera), a leafhopper with an XX/X0 sex-determination system.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1599}, pages = {2409-2416}, pmid = {16928646}, issn = {0962-8452}, mesh = {Animals ; Female ; Feminization/*microbiology ; Hemiptera/genetics/*microbiology/*physiology ; Male ; *Sex Determination Processes ; Wolbachia/*physiology ; }, abstract = {Zyginidia pullula is a grass-dwelling leafhopper characterized by a bisexual reproduction mode. In this species, some females collected in Northern Italy, when mated with males, gave origin to an exclusively female brood. Here, we demonstrated that in these lineages an infection by a new strain of Wolbachia pipientis-designated as wZygpul-was detected by amplifying and sequencing the wsp and 16S rRNA genes. About half of the female progeny were characterized by intersexual phenotypes, i.e. showing upper pygofer appendages, a typical male secondary sexual feature. The karyological analysis proved that while phenotypically normal females had a female genotype, those with upper pygofer appendages had a male genotype and were thus feminized males. The complete removal of W. pipientis after tetracycline treatment of morphologically normal females, and the consequent re-appearance of males in the brood, permitted us to connect the feminizing effect with the presence of the bacterium. This is the first case of feminization by W. pipientis in an XX/X0 sex-determination system, and is the second case reported in insects.}, } @article {pmid16924406, year = {2007}, author = {Panaram, K and Marshall, JL}, title = {F supergroup Wolbachia in bush crickets: what do patterns of sequence variation reveal about this supergroup and horizontal transfer between nematodes and arthropods?.}, journal = {Genetica}, volume = {130}, number = {1}, pages = {53-60}, doi = {10.1007/s10709-006-0020-7}, pmid = {16924406}, issn = {0016-6707}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; DNA, Ribosomal/analysis ; *Disease Transmission, Infectious ; *Genetic Variation ; Gryllidae/*microbiology ; Nematoda/*microbiology ; Phylogeny ; Wolbachia/*genetics ; }, abstract = {Wolbachia pipientis, an intracellular, alpha-proteobacterium, is commonly found in arthropods and filarial nematodes. Most infected insects are known to harbor strains of Wolbachia from supergroups A or B, whereas supergroups C and D occur only in filarial nematodes. Here, we present molecular evidence from two genes (ftsZ and 16S rDNA) that 2 Orthopterans (the bush cricket species Orocharis saltator and Hapithus agitator; Gryllidae: Eneopterinae) are infected with Wolbachia from the F supergroup. Additionally, a series of PCR tests revealed that these bush cricket specimens did not harbor nematodes, thus indicating that our positive results were not a by-product of nematodes being present in these cricket samples. Patterns of molecular variation suggest that (1) strains of F supergroup Wolbachia exhibit less genetic variation than the nematode-specific C and D supergroups but more than the A and B supergroups found in arthropods and (2) that there is no evidence of recombination within F supergroup strains. The above data support previous findings that F supergroup Wolbachia is not only harbored in both nematodes and arthropods, but that horizontal transfer has likely occurred recently between these diverse taxonomic groups (although the exact details of such horizontal transmissions remain unclear). Moreover, the limited genetic variation and lack of recombination in the F supergroup suggest that this clade of Wolbachia has radiated relatively rapidly with either (1) little time for recombination to occur or (2) selection against recombination as occurs in the mutualistic C and D strains of Wolbachia - both of which remain to be explored further.}, } @article {pmid16912076, year = {2006}, author = {Tram, U and Fredrick, K and Werren, JH and Sullivan, W}, title = {Paternal chromosome segregation during the first mitotic division determines Wolbachia-induced cytoplasmic incompatibility phenotype.}, journal = {Journal of cell science}, volume = {119}, number = {Pt 17}, pages = {3655-3663}, doi = {10.1242/jcs.03095}, pmid = {16912076}, issn = {0021-9533}, mesh = {Animals ; Chromatin/metabolism ; *Chromosome Segregation ; *Embryo, Nonmammalian/cytology/parasitology/physiology ; Female ; Male ; Mitosis/*physiology ; Phenotype ; Spindle Apparatus/metabolism ; *Wasps/embryology/genetics/parasitology/physiology ; *Wolbachia/cytology/physiology ; }, abstract = {The most common Wolbachia-induced phenotype in insects is cytoplasmic incompatibility (CI), which occurs when sperm from infected males fertilize eggs from uninfected females. CI produces distinct phenotypes in three closely related haplo-diploid species of the genus Nasonia: mortality in N. longicornis and N. giraulti, and conversion to male development in N. vitripennis. We demonstrate that the majority of CI-induced mortality occurs during embryogenesis and that the pattern of paternal chromosome segregation during the first mitosis is a good predictor of CI phenotype. In N. giraulti and N. longicornis, the paternal chromosomes mis-segregate, producing abnormal nuclei connected by chromatin bridges. Consequently, these embryos arrest development with very few and abnormal nuclei. In contrast, the paternal genome in N. vitripennis is either not segregated or mis-segregates to one of the two daughter nuclei. Consequently, these embryos continue development utilizing the maternally derived haploid nuclei, resulting in male offspring. The latter class is the first documented example of asymmetric mitotic segregation of abnormal chromosomes. We conclude that in haplo-diploids, CI-induced embryonic lethality occurs only when abnormal paternal genome segregation affects both products of the first mitotic division. This is the first study to associate differences in types of CI with specific cytological defects.}, } @article {pmid16911221, year = {2006}, author = {Duron, O and Bernard, C and Unal, S and Berthomieu, A and Berticat, C and Weill, M}, title = {Tracking factors modulating cytoplasmic incompatibilities in the mosquito Culex pipiens.}, journal = {Molecular ecology}, volume = {15}, number = {10}, pages = {3061-3071}, doi = {10.1111/j.1365-294X.2006.02996.x}, pmid = {16911221}, issn = {0962-1083}, mesh = {Animals ; Cell Nucleus/metabolism ; Crosses, Genetic ; Culex/*cytology/*metabolism ; Cytoplasm/*metabolism ; Female ; Genes, Insect/genetics ; Genome, Insect/genetics ; Geography ; Male ; Prophages ; }, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI), resulting in abortive embryonic development. One Wolbachia host, Culex pipiens complex mosquitoes, displays high levels of variability in both CI crossing types (cytotypes) and DNA markers. We report here an analysis of 14 mosquito strains, containing 13 Wolbachia variants, and with 13 different cytotypes. Cytotypes were Wolbachia-dependent, as antibiotic treatment rendered all strains tested compatible. Cytotype distributions were independent of geographical distance between sampling sites and host subspecies, suggesting that Wolbachia does not promote a reproductive isolation depending on these parameters. Backcross analysis demonstrated a mild restoring effect of the nuclear genome, indicating that CI is mostly cytoplasmically determined for some crosses. No correlation was found between the phenotypic and genotypic variability of 16 WO prophage and transposon markers, except for the WO prophage Gp15 gene, which encodes a protein similar to a bacterial virulence factor. However, Gp15 is partially correlated with CI expression, suggesting that it could be just linked to a CI gene.}, } @article {pmid16901827, year = {2006}, author = {Perlman, SJ and Hunter, MS and Zchori-Fein, E}, title = {The emerging diversity of Rickettsia.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1598}, pages = {2097-2106}, pmid = {16901827}, issn = {0962-8452}, mesh = {Animals ; *Biodiversity ; Invertebrates/*microbiology/physiology ; *Phylogeny ; Reproduction/physiology ; Rickettsia/*genetics/*pathogenicity ; }, abstract = {The best-known members of the bacterial genus Rickettsia are associates of blood-feeding arthropods that are pathogenic when transmitted to vertebrates. These species include the agents of acute human disease such as typhus and Rocky Mountain spotted fever. However, many other Rickettsia have been uncovered in recent surveys of bacteria associated with arthropods and other invertebrates; the hosts of these bacteria have no relationship with vertebrates. It is therefore perhaps more appropriate to consider Rickettsia as symbionts that are transmitted vertically in invertebrates, and secondarily as pathogens of vertebrates. In this review, we highlight the emerging diversity of Rickettsia species that are not associated with vertebrate pathogenicity. Phylogenetic analysis suggests multiple transitions between symbionts that are transmitted strictly vertically and those that exhibit mixed (horizontal and vertical) transmission. Rickettsia may thus be an excellent model system in which to study the evolution of transmission pathways. We also focus on the emergence of Rickettsia as a diverse reproductive manipulator of arthropods, similar to the closely related Wolbachia, including strains associated with male-killing, parthenogenesis, and effects on fertility. We emphasize some outstanding questions and potential research directions, and suggest ways in which the study of non-pathogenic Rickettsia can advance our understanding of their disease-causing relatives.}, } @article {pmid16900311, year = {2006}, author = {Groot, TV and Breeuwer, JA}, title = {Cardinium symbionts induce haploid thelytoky in most clones of three closely related Brevipalpus species.}, journal = {Experimental & applied acarology}, volume = {39}, number = {3-4}, pages = {257-271}, pmid = {16900311}, issn = {0168-8162}, mesh = {Animals ; Bacteroidetes/genetics/*physiology ; Female ; Male ; Mites/genetics/*physiology ; Parthenogenesis ; Reproduction ; *Symbiosis ; }, abstract = {Bacterial symbionts that manipulate the reproduction of their host to increase their own transmission are widespread. Most of these bacteria are Wolbachia, but recently a new bacterium, named Cardinium, was discovered that is capable of the same manipulations. In the host species Brevipalpus phoenicis (Acari: Tenuipalpidae) this bacterium induces thelytoky by feminizing unfertilized haploid eggs. The related species B. obovatus and B. californicus are thelytokous too, suggesting that they reproduce in the same remarkable way as B. phoenicis. Here we investigated the mode of thelytokous reproduction in these three species. Isofemale lines were created of all three species and 19 lines were selected based on variation in mitochondrial COI sequences. All B. phoenicis and B. californicus lines (10 and 4 lines, respectively) produced males under laboratory conditions up to 6.7%. In contrast, males were absent from all B. obovatus lines (5 lines). Additional experiments with two B. phoenicis isofemale lines showed that males can be produced by very young females only, while older females produce daughters exclusively. For most lines it was shown that they are indeed feminized by a bacterium as treatment with antibiotics resulted in increased numbers of males up to 13.5%. Amplification and identification of specific gyrB sequences confirmed that those lines were infected with Cardinium. Three out of the five B. obovatus lines did not produce males after treatments with antibiotics, nor did they contain Cardinium or any other bacterium that might induce thelytoky. In these lines thelytoky is probably a genetic property of the mite itself. Despite the different causes of thelytoky, flow cytometry revealed that all 19 lines were haploid. Finally, the taxonomic inferences based on the mitochondrial COI sequences were incongruent with the classical taxonomy based on morphology, suggesting that a taxonomic revision of this group is necessary.}, } @article {pmid16895998, year = {2006}, author = {Ruang-Areerate, T and Kittayapong, P}, title = {Wolbachia transinfection in Aedes aegypti: a potential gene driver of dengue vectors.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {103}, number = {33}, pages = {12534-12539}, pmid = {16895998}, issn = {0027-8424}, mesh = {*Aedes/genetics/microbiology ; Animals ; Dengue/*transmission ; Female ; *Insect Vectors ; Male ; *Rickettsiaceae Infections ; Symbiosis ; *Transgenes ; *Wolbachia/genetics/pathogenicity ; }, abstract = {The endosymbiotic bacteria in the genus Wolbachia are capable of inducing a wide range of reproductive abnormalities in their hosts, including cytoplasmic incompatibility (CI), which could lead to the replacement of uninfected host populations with infected ones. Because of this, Wolbachia have attracted considerable interest as a potential mechanism for spreading disease-blocking transgenes through vector populations. Here we report the establishment of double Wolbachia transinfection by direct adult microinjection of Wolbachia from naturally double-infected Aedes albopictus to Aedes aegypti, the most important mosquito vector of infectious viral diseases, and a mosquito in which natural Wolbachia infections are not known to occur. We further demonstrate that incomplete CI is induced in these double-transinfected mosquitoes. Comparisons of fitness traits between naturally uninfected and transinfected Ae. aegypti lines indicated one significant difference in favor of the latter, namely, an increased number of eggs laid. Levels of CI expression corresponded to the Wolbachia density. There were large differences in relative Wolbachia density between reproductive and nonreproductive tissues in both Ae. albopictus and transinfected Ae. aegypti, except Malpighian tubule, which implied the preferred establishment of Wolbachia within reproductive tissue. Results from a simulation model confirm that population replacement by transinfected Ae. aegypti is possible over time. The establishment of Wolbachia double infections in Ae. aegypti by direct adult microinjection and the demonstration of CI expression in this new host suggest that Wolbachia could be experimentally transferred into vector species and could also be used as a gene-driving system to genetically manipulate vector populations.}, } @article {pmid16892635, year = {2006}, author = {Tsai, KH and Huang, CG and Wu, WJ and Chuang, CK and Lin, CC and Chen, WJ}, title = {Parallel infection of Japanese encephalitis virus and Wolbachia within cells of mosquito salivary glands.}, journal = {Journal of medical entomology}, volume = {43}, number = {4}, pages = {752-756}, doi = {10.1603/0022-2585(2006)43[752:piojev]2.0.co;2}, pmid = {16892635}, issn = {0022-2585}, mesh = {Animals ; Culicidae/*microbiology/*virology ; Cytoplasm/microbiology ; DNA Primers/chemistry ; Encephalitis Virus, Japanese/*isolation & purification ; Female ; Fluorescent Antibody Technique, Indirect/methods ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Salivary Glands/microbiology/ultrastructure/virology ; Time Factors ; Wolbachia/*isolation & purification ; }, abstract = {The endosymbiont Wolbachia usually causes cytoplasmic incompatibility in dipteran hosts, including mosquitoes. However, some important arbovirus-transmitting mosquitoes such as Aedes aegypti (L.) are not heritably infected by Wolbachia. In Wolbachia-harboring mosquito Armigeres subalbatus Coquillett, colocalization of Wolbachia and inoculated Japanese encephalitis virus (family Flaviviridae, genus Flavivirus, JEV) in salivary gland (SG) cells was shown by electron microscopy. The infection rate of JEV in SGs, detected with either immunofluorescent antibody test or reverse transcription-polymerase chain reaction, did not show significant differences between Wolbachia-infected and -free colonies. It is suggested that Wolbachia did not mediate resistance of SG cells to superinfection by JEV, although both microorgamisms coexist in the same niche, i.e., the same SG cell. Therefore, a SG escape barrier may not be elevated due to Wolbachia infection, which presumably has no deleterious effects on vector competence in Wolbachia-harboring mosquitoes.}, } @article {pmid16892626, year = {2006}, author = {Sakamoto, JM and Rasgon, JL}, title = {Geographic distribution of Wolbachia infections in Cimex lectularius (Heteroptera: Cimicidae).}, journal = {Journal of medical entomology}, volume = {43}, number = {4}, pages = {696-700}, doi = {10.1603/0022-2585(2006)43[696:gdowii]2.0.co;2}, pmid = {16892626}, issn = {0022-2585}, mesh = {Animals ; Bedbugs/genetics/*microbiology ; DNA Primers/chemistry ; DNA, Bacterial/analysis ; DNA, Mitochondrial/analysis ; Female ; Geography ; Male ; North America ; Nymph/microbiology ; Pest Control, Biological/methods ; Polymerase Chain Reaction/methods ; Wolbachia/genetics/isolation & purification/*pathogenicity ; Zambia ; }, abstract = {Endosymbiotic Wolbachia bacteria have been previously shown to infect laboratory colonies of the human bed bug, Cimex lectularius L. (Heteroptera: Cimicidae), but little information exists regarding the extent of infection in natural populations. We assayed C. lectularius populations from five North American regions (California, Connecticut, Florida, New York, and Toronto, Canada) and one African region (Macha, Zambia) for Wolbachia infection by the polymerase chain reaction (PCR). Wolbachia infections were prevalent in all populations assayed (83-100%). There were no significant differences in infection frequency between geographic regions, between sexes, or between life stages (adult versus nymph). The potential utility of Wolbachia for alternative bed bug control strategies is discussed.}, } @article {pmid16892625, year = {2006}, author = {Islam, MS and Dobson, SL}, title = {Wolbachia effects on Aedes albopictus (Diptera: Culicidae) immature survivorship and development.}, journal = {Journal of medical entomology}, volume = {43}, number = {4}, pages = {689-695}, doi = {10.1603/0022-2585(2006)43[689:weoaad]2.0.co;2}, pmid = {16892625}, issn = {0022-2585}, support = {AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/growth & development/*microbiology/physiology ; Analysis of Variance ; Animal Nutritional Physiological Phenomena ; Animals ; Breeding ; Female ; Fertility/physiology ; Insect Vectors/growth & development/*microbiology/physiology ; Male ; Ovum/growth & development ; Survival Analysis ; Time Factors ; Wolbachia/*pathogenicity/physiology ; }, abstract = {Wolbachia bacteria manipulate the reproduction of mosquito hosts via a form of sterility known as cytoplasmic incompatibility (CI), promoting the spread of infections into host populations. The rate at which an infection invades is affected by host fitness costs associated with the Wolbachia infection. Here, we examine for an effect of Wolbachia infection on the immature fitness of the Asian tiger mosquito Aedes albopictus (Skuse) (Diptera: Culicidae). In two experiments, we examine for a Wolbachia effect on immature survivorship and developmental rate, adult size, and an effect of larval nutrition on CI level. The highest survivorship can be observed in uninfected larvae, primarily because of reduced survivorship of Wolbachia-infected males. Although differences in the developmental rates are observed between the examined strains, the differences cannot be readily attributed to Wolbachia. An effect of Wolbachia on adult size is not observed. Poor male nutrition is associated with reduced fecundity and egg hatch of mates. The latter is hypothesized to explain the reduced egg hatch observed in CI crosses of malnourished males relative to well fed males. We discuss the results in relation to previously identified differences in adult fitness, naturally occurring invasions of Wolbachia, applied strategies of population replacement, and the need for additional modeling effort.}, } @article {pmid16860767, year = {2006}, author = {Bisht, R and Hoti, SL and Thangadurai, R and Das, PK}, title = {Isolation of Wuchereria bancrofti microfilariae from archived stained blood slides for use in genetic studies and amplification of parasite and endosymbiont genes.}, journal = {Acta tropica}, volume = {99}, number = {1}, pages = {1-5}, doi = {10.1016/j.actatropica.2005.12.009}, pmid = {16860767}, issn = {0001-706X}, mesh = {Animals ; DNA, Helminth/chemistry/genetics ; Filariasis/*blood ; Genetic Variation ; Humans ; India ; Phylogeny ; Random Amplified Polymorphic DNA Technique ; Wuchereria bancrofti/*genetics/*isolation & purification ; }, abstract = {Information on change in genetic diversity of Wuchereria bancrofti is important in view of the launching of the Global Lymphatic Filariasis Elimination Programme, as it may have important consequences on the control operations and on the potential resurgence after their withdrawal. Since attention was not paid to generate such information when the programme was launched, use of archived parasite material will provide an opportunity to derive this information in a prospective manner. In this paper a simple and effective technique is reported for isolation of microfilariae of W. bancrofti from dried and stained slides archived for several years and their utility in analysis of genetic structure and amplification of certain genes of the parasite is tested. The method was found to be efficient in purifying mf from the dried smears and the DNA of the parasite found to be useful in studying the genetic structure of Wuchereria bancrofti populations using random amplified polymorphic DNA (RAPD)-PCR and for amplifying genes of the parasite and its endosymbiont, Wolbachia sp.}, } @article {pmid16850225, year = {2006}, author = {Montenegro, H and Petherwick, AS and Hurst, GD and Klaczko, LB}, title = {Fitness effects of Wolbachia and Spiroplasma in Drosophila melanogaster.}, journal = {Genetica}, volume = {127}, number = {1-3}, pages = {207-215}, doi = {10.1007/s10709-005-3766-4}, pmid = {16850225}, issn = {0016-6707}, mesh = {Animals ; Cell Survival ; Drosophila/*microbiology ; Extrachromosomal Inheritance ; Female ; Fertility ; Larva/growth & development ; Male ; Spiroplasma/*pathogenicity ; Symbiosis/*physiology ; Wolbachia/*pathogenicity ; }, abstract = {Maternally inherited endosymbionts that manipulate the reproduction of their insect host are very common. Aside from the reproductive manipulation they produce, the fitness of these symbionts depends in part on the direct impact they have on the female host. Although this parameter has commonly been investigated for single infections, it has much more rarely been established in dual infections. We here establish the direct effect of infection with two different symbionts exhibiting different reproductive manipulation phenotypes, both alone and in combination, in the fruit fly Drosophila melanogaster. This species carries a cytoplasmic incompatibility inducing Wolbachia and a male-killing Spiroplasma, occurring as single or double (co-) infections in natural populations. We assessed direct fitness effects of these bacteria on their host, by comparing larval competitiveness and adult fecundity of uninfected, Wolbachia, Spiroplasma and Wolbachia-Spiroplasma co-infected females. We found no effect of infection status on the fitness of females for both estimates, that is, no evidence of any benefits or costs to either single or co-infection. This leads to the conclusion that both bacteria probably have other sources of benefits to persist in D. melanogaster populations, either by means of their reproductive manipulations (fitness compensation from male death in Spiroplasma infection and cytoplasmic incompatibility in Wolbachia infection) or by positive fitness interactions on other fitness components.}, } @article {pmid16842419, year = {2006}, author = {Chauvatcharin, N and Ahantarig, A and Baimai, V and Kittayapong, P}, title = {Bacteriophage WO-B and Wolbachia in natural mosquito hosts: infection incidence, transmission mode and relative density.}, journal = {Molecular ecology}, volume = {15}, number = {9}, pages = {2451-2461}, doi = {10.1111/j.1365-294X.2006.02947.x}, pmid = {16842419}, issn = {0962-1083}, mesh = {Animals ; Bacteriophages/*genetics/*physiology ; Culicidae/classification/*genetics/*microbiology ; Incidence ; Microscopy, Electron, Transmission ; Molecular Sequence Data ; Phylogeny ; Rickettsiaceae Infections/microbiology/transmission/*veterinary ; Wolbachia/*physiology/*virology ; }, abstract = {Bacteriophages of Wolbachia bacteria have been proposed as a potential transformation tool for genetically modifying mosquito vectors. In this study, we report the presence of the WO-B class of Wolbachia-associated phages among natural populations of several mosquito hosts. Eighty-eight percent (22/25) of Wolbachia-infected mosquito species surveyed were found to contain WO-B phages. WO-B phage orf7 sequence analysis suggested that a single strain of WO-B phage was found in most singly (23/24) or doubly (1/1) Wolbachia-infected mosquitoes. However, the single Wolbachia strain infecting Aedes perplexus was found to harbour at least two different WO-B phages. Phylogenetic analysis suggested that horizontal transmission of WO-B phages has occurred on an evolutionary scale between the Wolbachia residing in mosquitoes. On an ecological scale, a low trend of co-transmission occurred among specific WO-B phages within Wolbachia of each mosquito species. Assessment of the density of WO-B phage by real-time quantitative polymerase chain reaction (RTQ-PCR) revealed an average relative density of 7.76 x 10(5)+/- 1.61 x 10(5) orf7 copies per individual mosquito for a single Wolbachia strain infecting mosquitoes, but a threefold higher density in the doubly Wolbachia-infected Aedes albopictus. However, the average combined density of WO-B phage(s) did not correlate with that of their Wolbachia hosts, which varied in different mosquito species. We also confirmed the presence of WO-B-like virus particles in the laboratory colony of Ae. albopictus (KLPP) morphologically, by transmission electron microscopy (TEM). The viral-like particles were detected after purification and filtration of Ae. albopictus ovary extract, suggesting that at least one WO-B-like phage is active (temperate) within the Wolbachia of this mosquito vector. Nevertheless, the idea of utilizing these bacteriophages as transformation vectors still needs more investigation and is likely to be unfeasible.}, } @article {pmid16820474, year = {2006}, author = {Goto, S and Anbutsu, H and Fukatsu, T}, title = {Asymmetrical interactions between Wolbachia and Spiroplasma endosymbionts coexisting in the same insect host.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {7}, pages = {4805-4810}, pmid = {16820474}, issn = {0099-2240}, mesh = {Animals ; DNA/analysis/isolation & purification ; DNA, Bacterial/analysis/isolation & purification ; Drosophila melanogaster/growth & development/*microbiology ; Female ; Polymerase Chain Reaction ; Spiroplasma/genetics/*growth & development ; *Symbiosis ; Wolbachia/genetics/*growth & development ; }, abstract = {We investigated the interactions between the endosymbionts Wolbachia pipientis strain wMel and Spiroplasma sp. strain NSRO coinfecting the host insect Drosophila melanogaster. By making use of antibiotic therapy, temperature stress, and hemolymph microinjection, we established the following strains in the same host genetic background: the SW strain, infected with both Spiroplasma and Wolbachia; the S strain, infected with Spiroplasma only; and the W strain, infected with Wolbachia only. The infection dynamics of the symbionts in these strains were monitored by quantitative PCR during host development. The infection densities of Spiroplasma exhibited no significant differences between the SW and S strains throughout the developmental course. In contrast, the infection densities of Wolbachia were significantly lower in the SW strain than in the W strain at the pupal and young adult stages. These results indicated that the interactions between the coinfecting symbionts were asymmetrical, i.e., Spiroplasma organisms negatively affected the population of Wolbachia organisms, while Wolbachia organisms did not influence the population of Spiroplasma organisms. In the host body, the symbionts exhibited their own tissue tropisms: among the tissues examined, Spiroplasma was the most abundant in the ovaries, while Wolbachia showed the highest density in Malpighian tubules. Strikingly, basically no Wolbachia organisms were detected in hemolymph, the principal location of Spiroplasma. These results suggest that different host tissues act as distinct microhabitats for the symbionts and that the lytic process in host metamorphosis might be involved in the asymmetrical interactions between the coinfecting symbionts.}, } @article {pmid16818783, year = {2006}, author = {Turner, JD and Langley, RS and Johnston, KL and Egerton, G and Wanji, S and Taylor, MJ}, title = {Wolbachia endosymbiotic bacteria of Brugia malayi mediate macrophage tolerance to TLR- and CD40-specific stimuli in a MyD88/TLR2-dependent manner.}, journal = {Journal of immunology (Baltimore, Md. : 1950)}, volume = {177}, number = {2}, pages = {1240-1249}, doi = {10.4049/jimmunol.177.2.1240}, pmid = {16818783}, issn = {0022-1767}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Adaptor Proteins, Signal Transducing/*physiology ; Animals ; Brugia malayi/*immunology/microbiology ; CD40 Antigens/biosynthesis/*physiology ; Cells, Cultured ; Dose-Response Relationship, Immunologic ; Down-Regulation/immunology ; Female ; *Immune Tolerance ; Lipopolysaccharide Receptors/biosynthesis ; Lipopolysaccharides/administration & dosage/pharmacology ; Macrophage Activation/immunology ; Macrophages, Peritoneal/*immunology/metabolism/microbiology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Mice, Knockout ; Myeloid Differentiation Factor 88 ; Symbiosis/*immunology ; Toll-Like Receptor 2/biosynthesis/deficiency/genetics/*physiology ; Toll-Like Receptor 4/antagonists & inhibitors/biosynthesis ; Transforming Growth Factor beta/biosynthesis ; Transforming Growth Factor beta1 ; Up-Regulation/immunology ; Wolbachia/*immunology ; }, abstract = {Lymphatic filarial nematodes are able to down-regulate parasite-specific and nonspecific responses of lymphocytes and APC. Lymphatic filariae are reliant on Wolbachia endosymbiotic bacteria for development and survival. We tested the hypothesis that repeated exposure to Wolbachia endosymbionts would drive macrophage tolerance in vitro and in vivo. We pre-exposed murine peritoneal-elicited macrophages to soluble extracts of Brugia malayi female worms (BMFE) before restimulating with BMFE or TLR agonists. BMFE tolerized macrophages (in terms of IFN-beta, IL-1beta, IL-6, IL-12p40, and TNF-alpha inflammatory cytokine production) in a dose-dependent manner toward self, LPS, MyD88-dependent TLR2 or TLR9 ligands (peptidoglycan, triacyl lipopeptide, CpG DNA) and the MyD88-independent/TRIF-dependent TLR3 ligand, polyinosinic-polycytidylic acid. This was accompanied with down-regulation in surface expression of TLR4 and up-regulation of CD14, CD40, and TLR2. BMFE tolerance extended to CD40 activation in vitro and systemic inflammation following lethal challenge in an in vivo model of endotoxin shock. The mechanism of BMFE-mediated macrophage tolerance was dependent on MyD88 and TLR2 but not TLR4. Evidence that desensitization was driven by Wolbachia-specific ligands was determined by use of extracts from Wolbachia-depleted B. malayi, aposymbiotic filarial species, and a cell line stably infected with Wolbachia pipientis. Our data promote a role for Wolbachia in contributing toward the dysregulated and tolerized immunological phenotype that accompanies the majority of human filarial infections.}, } @article {pmid16800120, year = {2006}, author = {Guerrero, R and Bain, O and Attout, T and Martin, C}, title = {The infective larva of Litomosoides yutajensis Guerrero et al., 2003 (Nematoda: Onchocercidae), a Wolbachia-free filaria from bat.}, journal = {Parasite (Paris, France)}, volume = {13}, number = {2}, pages = {127-130}, doi = {10.1051/parasite/2006132127}, pmid = {16800120}, issn = {1252-607X}, mesh = {Animals ; Arachnid Vectors/parasitology ; Chiroptera/*parasitology ; Female ; Filariasis/parasitology/transmission/*veterinary ; Filarioidea/anatomy & histology/*classification/growth & development ; Host-Parasite Interactions ; Larva ; *Life Cycle Stages ; Male ; Mites/parasitology ; Phylogeny ; Species Specificity ; Venezuela ; }, abstract = {The infective larva of Litomosoides yutajensis Guerrero et al., 2003, a parasite of the bat Pteronotus pamellii, is described; it is distinct from congeneric infective larvae by the absence of caudal lappets. The life cycles of five other species of Litomosoides are known; three are parasites of rodents, one of a marsupial and one of a bat. As with these species, the experimental vector of L. yutoajensis used was the macronyssid mite Ornithonyssus bacoti. In nature, the main vectors are probably other macronyssids but transmission by O. bacoti, with its large host-range, could account for the characteristic host-switchings in the evolution of Litomosoides. Unlike the murine model L. sigmodontis Chandler, 1931, L. yutajensis is devoid of the endosymbiontic bacteria Wolbachia and may be of great interest.}, } @article {pmid16783009, year = {2006}, author = {Mateos, M and Castrezana, SJ and Nankivell, BJ and Estes, AM and Markow, TA and Moran, NA}, title = {Heritable endosymbionts of Drosophila.}, journal = {Genetics}, volume = {174}, number = {1}, pages = {363-376}, pmid = {16783009}, issn = {0016-6731}, mesh = {Animals ; Bacterial Infections/*transmission ; DNA, Bacterial/analysis ; Drosophila/*genetics/*microbiology ; Female ; Gene Frequency ; Genetic Variation ; *Infectious Disease Transmission, Vertical ; Male ; Molecular Sequence Data ; Phylogeny ; Spiroplasma/genetics/physiology ; Symbiosis/*genetics ; Wolbachia/genetics/physiology ; }, abstract = {Although heritable microorganisms are increasingly recognized as widespread in insects, no systematic screens for such symbionts have been conducted in Drosophila species (the primary insect genetic models for studies of evolution, development, and innate immunity). Previous efforts screened relatively few Drosophila lineages, mainly for Wolbachia. We conducted an extensive survey of potentially heritable endosymbionts from any bacterial lineage via PCR screens of mature ovaries in 181 recently collected fly strains representing 35 species from 11 species groups. Due to our fly sampling methods, however, we are likely to have missed fly strains infected with sex ratio-distorting endosymbionts. Only Wolbachia and Spiroplasma, both widespread in insects, were confirmed as symbionts. These findings indicate that in contrast to some other insect groups, other heritable symbionts are uncommon in Drosophila species, possibly reflecting a robust innate immune response that eliminates many bacteria. A more extensive survey targeted these two symbiont types through diagnostic PCR in 1225 strains representing 225 species from 32 species groups. Of these, 19 species were infected by Wolbachia while only 3 species had Spiroplasma. Several new strains of Wolbachia and Spiroplasma were discovered, including ones divergent from any reported to date. The phylogenetic distribution of Wolbachia and Spiroplasma in Drosophila is discussed.}, } @article {pmid16777737, year = {2006}, author = {Champion de Crespigny, FE and Wedell, N}, title = {Wolbachia infection reduces sperm competitive ability in an insect.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1593}, pages = {1455-1458}, pmid = {16777737}, issn = {0962-8452}, mesh = {Animals ; Drosophila/*microbiology/physiology ; Fertility ; Male ; Spermatozoa/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {The maternally inherited bacterium Wolbachia pipientis imposes significant fitness costs on its hosts. One such cost is decreased sperm production resulting in reduced fertility of male Drosophila simulans infected with cytoplasmic incompatibility (CI) inducing Wolbachia. We tested the hypothesis that Wolbachia infection affects sperm competitive ability and found that Wolbachia infection is indeed associated with reduced success in sperm competition in non-virgin males. In the second male role, infected males sired 71% of the offspring whereas uninfected males sired 82% of offspring. This is the first empirical evidence indicating that Wolbachia infection deleteriously affects sperm competition and raises the possibility that polyandrous females can utilize differential sperm competitive ability to bias the paternity of broods and avoid the selfish manipulations of Wolbachia. This suggests a relationship between Wolbachia infection and host reproductive strategies. These findings also have important consequences for Wolbachia population dynamics because the transmission advantage of Wolbachia is likely to be undermined by sperm competition.}, } @article {pmid16777731, year = {2006}, author = {Dean, MD}, title = {A Wolbachia-associated fitness benefit depends on genetic background in Drosophila simulans.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1592}, pages = {1415-1420}, pmid = {16777731}, issn = {0962-8452}, mesh = {Animals ; Competitive Behavior ; Drosophila/*genetics/*microbiology/physiology ; Fertility ; Microinjections ; Wolbachia/*physiology ; }, abstract = {The alpha-proteobacteria Wolbachia infect a number of insect species and influence host reproduction to favour the spread of infected females through a population. The fitness effect of this infection is important in understanding the spread and maintenance of Wolbachia within and among host populations. However, a full elucidation of fitness effect requires careful control of host genetic background. Here, I transferred a single clone of Wolbachia (the wHa strain) into three genetically distinct isofemale lines of the fly Drosophila simulans using microinjection methodology. These lines carried one of the three described mitochondrial haplogroups (siI, siII or siIII) and differ in nuclear genome as well. Population cage assays showed that wHa-infected siIII flies enjoyed a dramatic fitness benefit compared to uninfected siIII. In contrast, wHa did not affect the fitness of siI or siII flies. This study points to the importance of host-by-symbiont interaction terms that may play an important role in organismal-fitness.}, } @article {pmid16777718, year = {2006}, author = {Xi, Z and Khoo, CC and Dobson, SL}, title = {Interspecific transfer of Wolbachia into the mosquito disease vector Aedes albopictus.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1592}, pages = {1317-1322}, pmid = {16777718}, issn = {0962-8452}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; AI 51533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology ; Animals ; Drosophila/microbiology ; Female ; Fertility ; Insect Vectors/*microbiology ; Male ; Microinjections ; Oocytes/cytology/microbiology ; Population Control/methods ; Rickettsiaceae Infections/*prevention & control/transmission ; Wolbachia/genetics/*physiology ; }, abstract = {Intracellular Wolbachia bacteria are obligate, maternally inherited endosymbionts found frequently in insects and other invertebrates. The evolutionary success of Wolbachia is due in part to an ability to manipulate reproduction. In mosquitoes and many other insects, Wolbachia causes a form of sterility known as cytoplasmic incompatibility (CI). Wolbachia-induced CI has attracted interest as a potential agent for affecting medically important disease vectors. However, application of the approach has been restricted by an absence of appropriate, naturally occurring Wolbachia infections. Here, we report the interspecific transfer of Wolbachia infection into a medically important mosquito. Using embryonic microinjection, Wolbachia is transferred from Drosophila simulans into the invasive pest and disease vector: Aedes albopictus (Asian tiger mosquito). The resulting infection is stably maintained and displays a unique pattern of bidirectional CI in crosses with naturally infected mosquitoes. Laboratory population cage experiments examine a strategy in which releases of Wolbachia-infected males are used to suppress mosquito egg hatch. We discuss the results in relation to developing appropriate Wolbachia-infected mosquito strains for population replacement and population suppression strategies.}, } @article {pmid16769630, year = {2006}, author = {Rasgon, JL and Cornel, AJ and Scott, TW}, title = {Evolutionary history of a mosquito endosymbiont revealed through mitochondrial hitchhiking.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1594}, pages = {1603-1611}, pmid = {16769630}, issn = {0962-8452}, support = {F31 GM020092/GM/NIGMS NIH HHS/United States ; GM020092/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Culex/*genetics/microbiology ; DNA, Mitochondrial/*chemistry ; *Evolution, Molecular ; *Genes, Mitochondrial ; Genetic Variation ; Haplotypes ; Phylogeny ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Due to cytoplasmic inheritance, spread of maternally inherited Wolbachia symbionts can result in reduction of mitochondrial variation in populations. We examined sequence diversity of the mitochondrial NADH dehydrogenase subunit 4 (ND4) gene in Wolbachia-infected (South Africa (SA), California and Thailand) and uninfected (SA) Culex pipiens complex populations. In total, we identified 12 haplotypes (A-L). In infected populations, 99% of individuals had haplotype K. In the uninfected SA population, 11 haplotypes were present, including K. Nuclear allozyme diversity was similar between infected and uninfected SA populations. Analysis of nuclear DNA sequences suggested that haplotype K presence in uninfected SA Cx. pipiens was probably due to a shared ancestral polymorphism rather than hybrid introgression. These data indicate that Wolbachia spread has resulted in drastic reduction of mitochondrial variability in widely separated Cx. pipiens complex populations. In contrast, the uninfected SA population is probably a cryptic species where Wolbachia introgression has been prevented by reproductive isolation, maintaining ancestral levels of mitochondrial diversity. Molecular clock analyses suggest that the Wolbachia sweep occurred within the last 47000 years. The effect of Wolbachia on mitochondrial dynamics can provide insight on the potential for Wolbachia to spread transgenes into mosquito populations to control vector-borne diseases.}, } @article {pmid16768810, year = {2006}, author = {Jaramillo, J and Borgemeister, C and Baker, P}, title = {Coffee berry borer Hypothenemus hampei (Coleoptera: Curculionidae): searching for sustainable control strategies.}, journal = {Bulletin of entomological research}, volume = {96}, number = {3}, pages = {223-233}, doi = {10.1079/ber2006434}, pmid = {16768810}, issn = {0007-4853}, mesh = {Agriculture/methods/*trends ; Animals ; Coffee/*parasitology ; Geography ; *Insect Control ; Nematoda/pathogenicity/physiology ; *Pest Control, Biological ; Weevils/microbiology/parasitology/*physiology ; Wolbachia/pathogenicity/physiology ; }, abstract = {The coffee berry borer Hypothenemus hampei (Ferrari) is the most serious pest of the world's most valuable tropical export crop. Since the last review on this insect was published six years ago, many new studies have contributed to an improved insight into the biology and ecology of the beetle, and have indicated new avenues for integrated and biological control. The latest developments in research, both laboratory and field, on the pest, its natural enemies and their implications for integrated control of H. hampei are summarized, with a particular focus on the situation in The Americas. Lately, the global coffee industry has changed radically; it has suffered a long cycle of lowest-ever world market prices caused by overproduction and technological change. At the same time, the advent of sustainable certification schemes has had a major impact on the industry. The role of integrated pest management and biological control of H. hampei in an era of changes in the coffee industry is discussed.}, } @article {pmid16739448, year = {2006}, author = {Wade, MJ and Goodnight, CJ}, title = {Cyto-nuclear epistasis: two-locus random genetic drift in hermaphroditic and dioecious species.}, journal = {Evolution; international journal of organic evolution}, volume = {60}, number = {4}, pages = {643-659}, pmid = {16739448}, issn = {0014-3820}, support = {1R01GM 065414-01A1/GM/NIGMS NIH HHS/United States ; }, mesh = {Alleles ; Animals ; Cell Nucleus/metabolism ; Cytoplasm/metabolism ; *Epistasis, Genetic ; Evolution, Molecular ; Female ; Genetic Drift ; Genetic Variation ; Genome ; Male ; Models, Genetic ; Wolbachia ; }, abstract = {We report the findings of our theoretical investigation of the effect of random genetic drift on the covariance of identity-by-descent (ibd) of nuclear and cytoplasmic genes. The covariance in ibd measures of the degree to which cyto-nuclear gene combinations are heritable, that is, transmitted together from parents to offspring. We show how the mating system affects the covariance of ibd, a potentially important aspect of host-pathogen or host-symbiont coevolution. The magnitude of this covariance influences the degree to which the evolution of apparently neutral cytoplasmic genes, often used in molecular phylogenetics, might be influenced by selection acting on unlinked nuclear genes. To the extent that cyto-nuclear gene combinations are inherited together, genomic conflict is mitigated and intergenomic transfer it facilitated, because genes in both organelle and nuclear genomes share the same evolutionary fate. The covariance of ibd also affects the rate at which cyto-nuclear epistatic variance is converted to additive variance necessary for a response to selection. We find that conversion is biased in species with separate sexes, so that the increment of additive variance added to the nuclear genome exceeds that added to the cytoplasmic genome. As a result, the host might have an adaptive advantage in a coevolutionary arms race with vertically (maternally) transmitted pathogens. Similarly, the nuclear genome could be a source of compensatory mutations for its organellar genomes, as occurs in cytoplasmic male sterility in some plant species. We also discuss the possibility that adaptive cytoplasmic elements, such as favorable mitochondrial mutations or endosymbionts (e.g., Wolbachia), have the potential to release heritable nuclear variation as they sweep through a host population, supporting the view that cytoplasmic introgression plays an important role in adaptation and speciation.}, } @article {pmid16724067, year = {2006}, author = {Frydman, HM and Li, JM and Robson, DN and Wieschaus, E}, title = {Somatic stem cell niche tropism in Wolbachia.}, journal = {Nature}, volume = {441}, number = {7092}, pages = {509-512}, doi = {10.1038/nature04756}, pmid = {16724067}, issn = {1476-4687}, mesh = {Abdomen/microbiology ; Animals ; Disease Transmission, Infectious ; Drosophila melanogaster/*microbiology ; Female ; Germ Cells/*microbiology ; Host-Parasite Interactions ; Infectious Disease Transmission, Vertical ; Organ Specificity ; Stem Cells/cytology/metabolism/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular bacteria found in the reproductive tissue of all major groups of arthropods. They are transmitted vertically from the female hosts to their offspring, in a pattern analogous to mitochondria inheritance. But Wolbachia phylogeny does not parallel that of the host, indicating that horizontal infectious transmission must also occur. Insect parasitoids are considered the most likely vectors, but the mechanism for horizontal transfer is largely unknown. Here we show that newly introduced Wolbachia cross several tissues and infect the germline of the adult Drosophila melanogaster female. Through investigation of bacterial migration patterns during the course of infection, we found that Wolbachia reach the germline through the somatic stem cell niche in the D. melanogaster germarium. In addition, our data suggest that Wolbachia are highly abundant in the somatic stem cell niche of long-term infected hosts, implying that this location may also contribute to efficient vertical transmission. This is, to our knowledge, the first report of an intracellular parasite displaying tropism for a stem cell niche.}, } @article {pmid16714198, year = {2006}, author = {Enk, CD}, title = {Onchocerciasis--river blindness.}, journal = {Clinics in dermatology}, volume = {24}, number = {3}, pages = {176-180}, doi = {10.1016/j.clindermatol.2005.11.008}, pmid = {16714198}, issn = {0738-081X}, mesh = {Humans ; *Onchocerciasis, Ocular/diagnosis/drug therapy ; }, abstract = {Onchocerciasis results from infestation by the nematode Onchocerca volvulus and is characterized by troublesome itching, skin lesions, and eye manifestations. Although partially controlled by international mass prevention programs, onchocerciasis remains a major health hazard and is endemic in Africa, Arabia, and the Americas. Onchocerciasis is spread by bites from infested black flies, which transmit larvae that subsequently develop into adult filariae. Skin symptoms are commonly nonspecific and include severe pruritus, acute and chronic dermatitis, vitiligo-like hypopigmentation, and atrophy. Onchocercal ocular disease covers a large spectrum of manifestations, which in severe cases, may lead to blindness. Diagnosis is usually made by direct visualization of the larvae emerging from superficial skin biopsies, "skin snips." In some cases, the microfilariae can also be directly observed at the slit lamp when migrating into the anterior chamber of the eye. Ivermectin is, at present, the drug of choice for skin and ocular manifestations. Recent research using a chemotherapeutic approach that targets filarial Wolbachia symbionts in the treatment and control of onchocerciasis, however, suggests that 100 mg/d of doxycycline for 6 weeks might be effective in reducing the filarial load and preventing ocular symptoms.}, } @article {pmid16710453, year = {2006}, author = {Bordenstein, SR and Marshall, ML and Fry, AJ and Kim, U and Wernegreen, JJ}, title = {The tripartite associations between bacteriophage, Wolbachia, and arthropods.}, journal = {PLoS pathogens}, volume = {2}, number = {5}, pages = {e43}, pmid = {16710453}, issn = {1553-7374}, support = {R01 GM062626/GM/NIGMS NIH HHS/United States ; R01 GM62626-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/growth & development/*physiology ; Female ; Gene Dosage ; Gram-Negative Bacterial Infections/microbiology ; Male ; Penetrance ; Spermatids/physiology ; Symbiosis/*physiology ; Wasps/microbiology/*physiology ; Wolbachia/genetics/growth & development/*physiology ; }, abstract = {By manipulating arthropod reproduction worldwide, the heritable endosymbiont Wolbachia has spread to pandemic levels. Little is known about the microbial basis of cytoplasmic incompatibility (CI) except that bacterial densities and percentages of infected sperm cysts associate with incompatibility strength. The recent discovery of a temperate bacteriophage (WO-B) of Wolbachia containing ankyrin-encoding genes and virulence factors has led to intensifying debate that bacteriophage WO-B induces CI. However, current hypotheses have not considered the separate roles that lytic and lysogenic phage might have on bacterial fitness and phenotype. Here we describe a set of quantitative approaches to characterize phage densities and its associations with bacterial densities and CI. We enumerated genome copy number of phage WO-B and Wolbachia and CI penetrance in supergroup A- and B-infected males of the parasitoid wasp Nasonia vitripennis. We report several findings: (1) variability in CI strength for A-infected males is positively associated with bacterial densities, as expected under the bacterial density model of CI, (2) phage and bacterial densities have a significant inverse association, as expected for an active lytic infection, and (3) CI strength and phage densities are inversely related in A-infected males; similarly, males expressing incomplete CI have significantly higher phage densities than males expressing complete CI. Ultrastructural analyses indicate that approximately 12% of the A Wolbachia have phage particles, and aggregations of these particles can putatively occur outside the Wolbachia cell. Physical interactions were observed between approximately 16% of the Wolbachia cells and spermatid tails. The results support a low to moderate frequency of lytic development in Wolbachia and an overall negative density relationship between bacteriophage and Wolbachia. The findings motivate a novel phage density model of CI in which lytic phage repress Wolbachia densities and therefore reproductive parasitism. We conclude that phage, Wolbachia, and arthropods form a tripartite symbiotic association in which all three are integral to understanding the biology of this widespread endosymbiosis. Clarifying the roles of lytic and lysogenic phage development in Wolbachia biology will effectively structure inquiries into this research topic.}, } @article {pmid16701248, year = {2004}, author = {Fenn, K and Blaxter, M}, title = {Are filarial nematode Wolbachia obligate mutualist symbionts?.}, journal = {Trends in ecology & evolution}, volume = {19}, number = {4}, pages = {163-166}, doi = {10.1016/j.tree.2004.01.002}, pmid = {16701248}, issn = {0169-5347}, abstract = {The intracellular symbiotic bacteria of filarial nematodes have inspired new ideas for the control of disease using antibacterial drugs. For effective, long-term control, this requires that the bacteria are essential to their nematode hosts. Two recent studies offer conflicting evidence: long, close coevolution between most filarial nematodes and their symbionts contrasts with many species having naturally lost them. An attempt to transfer symbionts to an uninfected host found that the bacteria did not thrive, suggesting they are adapted to one host.}, } @article {pmid16687688, year = {2006}, author = {Rao, RU and Atkinson, LJ and Ramzy, RM and Helmy, H and Farid, HA and Bockarie, MJ and Susapu, M and Laney, SJ and Williams, SA and Weil, GJ}, title = {A real-time PCR-based assay for detection of Wuchereria bancrofti DNA in blood and mosquitoes.}, journal = {The American journal of tropical medicine and hygiene}, volume = {74}, number = {5}, pages = {826-832}, pmid = {16687688}, issn = {0002-9637}, support = {U01 AI035855/AI/NIAID NIH HHS/United States ; U01 AI035855-09/AI/NIAID NIH HHS/United States ; U01 AI035855-06/AI/NIAID NIH HHS/United States ; U01 AI035855-10S1/AI/NIAID NIH HHS/United States ; U01 AI035855-07/AI/NIAID NIH HHS/United States ; U01 AI035855-10/AI/NIAID NIH HHS/United States ; U01 AI035855-08/AI/NIAID NIH HHS/United States ; AI 35855/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*parasitology ; Culex/*parasitology ; DNA Primers ; DNA, Bacterial/analysis ; DNA, Protozoan/analysis ; Egypt ; Filariasis/blood/*diagnosis/transmission ; Humans ; Insect Vectors/parasitology ; Papua New Guinea ; Polymerase Chain Reaction ; Predictive Value of Tests ; RNA, Ribosomal, 16S/analysis ; Reagent Kits, Diagnostic ; Sensitivity and Specificity ; Wolbachia/genetics/isolation & purification ; Wuchereria bancrofti/*genetics/isolation & purification ; }, abstract = {We developed and evaluated real-time polymerase chain reaction (PCR) assays for detecting Wuchereria bancrofti DNA in human blood and in mosquitoes. An assay based on detection of the W. bancrofti "LDR" repeat DNA sequence was more sensitive than an assay for Wolbachia 16S rDNA. The LDR-based assay was sensitive for detecting microfilarial DNA on dried membrane filters or on filter paper. We also compared real-time PCR with conventional PCR (C-PCR) for detecting W. bancrofti DNA in mosquito samples collected in endemic areas in Egypt and Papua New Guinea. Although the two methods had comparable sensitivity for detecting filarial DNA in reference samples, real-time PCR was more sensitive than C-PCR in practice with field samples. Other advantages of real-time PCR include its high-throughput capacity and decreased risk of cross-contamination between test samples. We believe that real-time PCR has great potential as a tool for monitoring progress in large-scale filariasis elimination programs.}, } @article {pmid16685282, year = {2006}, author = {Pool, JE and Wong, A and Aquadro, CF}, title = {Finding of male-killing Spiroplasma infecting Drosophila melanogaster in Africa implies transatlantic migration of this endosymbiont.}, journal = {Heredity}, volume = {97}, number = {1}, pages = {27-32}, pmid = {16685282}, issn = {0018-067X}, support = {R01 GM036431/GM/NIGMS NIH HHS/United States ; R01 GM036431-14/GM/NIGMS NIH HHS/United States ; GM36431/GM/NIGMS NIH HHS/United States ; }, mesh = {Aging ; Animal Migration ; Animals ; Drosophila melanogaster/*genetics/*microbiology/physiology ; Female ; Male ; Reproduction ; Sex Ratio ; Spiroplasma/*genetics/isolation & purification/physiology ; Symbiosis ; Uganda ; Wolbachia/genetics/isolation & purification/physiology ; }, abstract = {We report the identification of male-killing Spiroplasma in a wild-caught female Drosophila melanogaster from Uganda, the first such infection to be found in this species outside of South America. Among 38 female flies collected from Namulonge, Uganda in April, 2005, one produced a total of 41 female offspring but no males. PCR testing of subsequent generations revealed that females retaining Spiroplasma infection continued to produce a large excess of female progeny, while females that had lost Spiroplasma produced offspring with normal sex ratios. Consistent with earlier work, we find that male-killing and transmission efficiency appear to increase with female age, and we note that males born in sex ratio broods display much lower survivorship than their female siblings. DNA sequence comparisons at three loci suggest that this Spiroplasma strain is closely related to the male-killing strain previously found to infect D. melanogaster in Brazil, although part of one locus appears to show a recombinant history. Implications for the origin and history of male-killing Spiroplasma in D. melanogaster are discussed.}, } @article {pmid16682981, year = {2006}, author = {Sinkins, SP and Gould, F}, title = {Gene drive systems for insect disease vectors.}, journal = {Nature reviews. Genetics}, volume = {7}, number = {6}, pages = {427-435}, doi = {10.1038/nrg1870}, pmid = {16682981}, issn = {1471-0056}, mesh = {Animals ; Culicidae/*genetics/*growth & development ; DNA Transposable Elements ; *Disease Vectors ; Genetic Engineering ; Humans ; *Insect Vectors/genetics/growth & development ; Insecta ; Models, Genetic ; Organisms, Genetically Modified/*growth & development ; Parasitic Diseases/*prevention & control ; Wolbachia/genetics/growth & development ; }, abstract = {The elegant mechanisms by which naturally occurring selfish genetic elements, such as transposable elements, meiotic drive genes, homing endonuclease genes and Wolbachia, spread at the expense of their hosts provide some of the most fascinating and remarkable subjects in evolutionary genetics. These elements also have enormous untapped potential to be used in the control of some of the world's most devastating diseases. Effective gene drive systems for spreading genes that can block the transmission of insect-borne pathogens are much needed. Here we explore the potential of natural gene drive systems and discuss the artificial constructs that could be envisaged for this purpose.}, } @article {pmid16672513, year = {2006}, author = {Gottlieb, Y and Ghanim, M and Chiel, E and Gerling, D and Portnoy, V and Steinberg, S and Tzuri, G and Horowitz, AR and Belausov, E and Mozes-Daube, N and Kontsedalov, S and Gershon, M and Gal, S and Katzir, N and Zchori-Fein, E}, title = {Identification and localization of a Rickettsia sp. in Bemisia tabaci (Homoptera: Aleyrodidae).}, journal = {Applied and environmental microbiology}, volume = {72}, number = {5}, pages = {3646-3652}, pmid = {16672513}, issn = {0099-2240}, mesh = {Animals ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Glutamate Synthase/genetics ; Hemiptera/*growth & development/*microbiology ; In Situ Hybridization, Fluorescence ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*classification/genetics/*isolation & purification ; Sequence Analysis, DNA ; }, abstract = {Whiteflies (Homoptera: Aleyrodidae) are sap-sucking insects that harbor "Candidatus Portiera aleyrodidarum," an obligatory symbiotic bacterium which is housed in a special organ called the bacteriome. These insects are also home for a diverse facultative microbial community which may include Hamiltonella, Arsenophonus, Fritchea, Wolbachia, and Cardinium spp. In this study, the bacteria associated with a B biotype of the sweet potato whitefly Bemisia tabaci were characterized using molecular fingerprinting techniques, and a Rickettsia sp. was detected for the first time in this insect family. Rickettsia sp. distribution, transmission and localization were studied using PCR and fluorescence in situ hybridizations (FISH). Rickettsia was found in all 20 Israeli B. tabaci populations screened but not in all individuals within each population. A FISH analysis of B. tabaci eggs, nymphs, and adults revealed a unique concentration of Rickettsia around the gut and follicle cells, as well as a random distribution in the hemolymph. We postulate that the Rickettsia enters the oocyte together with the bacteriocytes, leaves these symbiont-housing cells when the egg is laid, multiplies and spreads throughout the egg during embryogenesis and, subsequently, disperses throughout the body of the hatching nymph, excluding the bacteriomes. Although the role Rickettsia plays in the biology of the whitefly is currently unknown, the vertical transmission on the one hand and the partial within-population infection on the other suggest a phenotype that is advantageous under certain conditions but may be deleterious enough to prevent fixation under others.}, } @article {pmid16672453, year = {2006}, author = {Sakamoto, JM and Feinstein, J and Rasgon, JL}, title = {Wolbachia infections in the Cimicidae: museum specimens as an untapped resource for endosymbiont surveys.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {5}, pages = {3161-3167}, pmid = {16672453}, issn = {0099-2240}, mesh = {Animals ; Cimicidae/classification/*microbiology ; DNA, Bacterial/analysis/isolation & purification ; Molecular Sequence Data ; *Museums ; Phylogeny ; Polymerase Chain Reaction ; Prevalence ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/genetics/*isolation & purification/*pathogenicity ; }, abstract = {Wolbachia spp. are obligate maternally inherited endosymbiotic bacteria that infect diverse arthropods and filarial nematodes. Previous microscopic and molecular studies have identified Wolbachia in several bed bug species (Cimicidae), but little is known about how widespread Wolbachia infections are among the Cimicidae. Because cimicids of non-medical importance are not commonly collected, we hypothesized that preserved museum specimens could be assayed for Wolbachia infections. For the screening of museum specimens, we designed a set of primers that specifically amplify small diagnostic fragments (130 to 240 bp) of the Wolbachia 16S rRNA gene. Using these and other previously published primers, we screened 39 cimicid species (spanning 16 genera and all 6 recognized subfamilies) and 2 species of the sister family Polyctenidae for Wolbachia infections using museum and wild-caught material. Amplified fragments were sequenced to confirm that our primers were amplifying Wolbachia DNA. We identified 10 infections, 8 of which were previously undescribed. Infections in the F supergroup were common in the subfamily Cimicinae, while infections in the A supergroup were identified in the subfamilies Afrocimicinae and Haematosiphoninae. Even though specimens were degraded, we detected infections in over 23% of cimicid species. Our results indicate that Wolbachia infections may be common among cimicids and that archived museum material is a useful untapped resource for invertebrate endosymbiont surveys. The new screening primers listed in this report will be useful for other researchers conducting Wolbachia surveys with specimens with less-than-optimum DNA quality.}, } @article {pmid16639421, year = {2006}, author = {Duron, O and Weill, M}, title = {Wolbachia infection influences the development of Culex pipiens embryo in incompatible crosses.}, journal = {Heredity}, volume = {96}, number = {6}, pages = {493-500}, doi = {10.1038/sj.hdy.6800831}, pmid = {16639421}, issn = {0018-067X}, mesh = {Animals ; Chromosome Mapping ; Crosses, Genetic ; Culex/*embryology/*microbiology ; DNA Primers ; Embryo, Nonmammalian/*microbiology ; Female ; Geography ; Male ; Polymerase Chain Reaction ; Reproduction ; Wolbachia/*pathogenicity/physiology ; X Chromosome ; Y Chromosome ; }, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and have evolved several different ways for manipulating their host, the most frequent being cytoplasmic incompatibility (CI). CI leads to embryo death in crosses between infected males and uninfected females, as well as in crosses between individuals infected by incompatible Wolbachia strains. In the mosquito Culex pipiens, previous studies suggested developmental variation in embryos stemming from different incompatible crosses. We have investigated this variation in different incompatible crosses. Unhatched eggs were separated into three classes based upon the developmental stage reached by the embryos. We found that incompatible crosses involving uninfected females produced only embryos whose development was arrested at a very early stage, irrespective of the Wolbachia variant infecting the male. These results differ from other host species where a developmental gradient that could reach late stages of embryogenesis or even living larvae was observed, and indicate a novel peculiarity of CI mechanism in C. pipiens. By contrast, all incompatible crosses with infected C. pipiens females produced embryos of all three classes. The proportion of embryo classes appeared to be associated with the strains involved, suggesting specific CI properties in different incompatible crosses. In addition, the contribution of parental genome was characterized in embryo classes using molecular markers for each chromosome. Embryo phenotypes appeared linked to the paternal chromosomes' contribution, as described in Drosophila simulans. However, this contribution varied according to maternal infection and independently of male factors.}, } @article {pmid16627277, year = {2006}, author = {Engelstädter, J and Charlat, S}, title = {Outbreeding selects for spiteful cytoplasmic elements.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1589}, pages = {923-929}, pmid = {16627277}, issn = {0962-8452}, mesh = {Animals ; *Breeding ; Extrachromosomal Inheritance/*physiology ; Female ; Infertility, Male/genetics ; Male ; Population Dynamics ; *Sexual Behavior, Animal ; Wolbachia/genetics ; }, abstract = {In addition to their nuclear genome, the vast majority of eukaryotes harbour cytoplasmic genomes, e.g. in mitochondria or chloroplasts. In the majority of cases, these cytoplasmic genomes are transmitted maternally only, leading to selective pressures divergent from those that act on nuclear genes. In particular, cytoplasmic genes, which reduce the fitness of males that carry them, but have no fitness effect in females, are believed to be selectively neutral. Here, we go a step further and argue that in outbreeding populations (i.e. populations with inbreeding avoidance), 'spiteful' cytoplasmic elements that reduce the number of offspring produced by males are in fact selected for. We study this process by means of a stochastic model, analysing both the probability of spread and the impact that such a spiteful cytotype can have on population dynamics. Our results demonstrate that the probability of spread of the spiteful cytotype can be several times higher in outbreeding than in panmictic populations. Spread and fixation of the spiteful cytotype can lead to different qualitative effects on the population dynamics, including extinction, decreased or increased stable population size. We discuss our results in respect to cytoplasmically induced male infertility and cytoplasmic incompatibility.}, } @article {pmid16624919, year = {2006}, author = {Ferree, PM and Sullivan, W}, title = {A genetic test of the role of the maternal pronucleus in Wolbachia-induced cytoplasmic incompatibility in Drosophila melanogaster.}, journal = {Genetics}, volume = {173}, number = {2}, pages = {839-847}, pmid = {16624919}, issn = {0016-6731}, mesh = {Animals ; Base Sequence ; Cell Nucleus/genetics ; Crosses, Genetic ; Cytoplasm/microbiology ; DNA/genetics ; Drosophila melanogaster/*genetics/*microbiology/physiology ; Female ; Male ; Models, Genetic ; Reproduction ; Wolbachia/*pathogenicity ; }, abstract = {Cytoplasmic incompatibility (CI) is a reproductive sterility found in arthropods that is caused by the endoparasitic bacteria Wolbachia. In CI, host progeny fail to develop during early embryogenesis if Wolbachia-infected males fertilize uninfected females. It is widely accepted that this lethality is caused by some unknown Wolbachia-induced modification of the paternal nuclear material in the host testes. However, the direct means by which this modification leads to early embryonic death are currently unresolved. Results from previous studies suggested that CI lethality occurs as a result of asynchrony in cell cycle timing between the paternal and maternal pronuclei. This hypothesis can be tested experimentally by the prediction that the Wolbachia-modified paternal pronucleus should support androgenetic development (i.e., from the paternal pronucleus only). Using specific mutations in Drosophila melanogaster that produce androgenetic progeny, we demonstrate that the Wolbachia-induced modification inhibits this type of development. This result suggests that CI occurs independently of the maternal pronucleus and argues against pronuclear asynchrony as the primary cause of CI lethality. We propose that CI occurs instead as the result of either a developmentally incompetent paternal pronucleus or asynchrony between the paternal pronucleus and the cell cycle of the egg cytoplasm.}, } @article {pmid16618673, year = {2006}, author = {Rio, RV and Wu, YN and Filardo, G and Aksoy, S}, title = {Dynamics of multiple symbiont density regulation during host development: tsetse fly and its microbial flora.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1588}, pages = {805-814}, pmid = {16618673}, issn = {0962-8452}, support = {R01 AI051584/AI/NIAID NIH HHS/United States ; T32 AI007404/AI/NIAID NIH HHS/United States ; AI34033/AI/NIAID NIH HHS/United States ; T32AI07404/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Enterobacteriaceae/isolation & purification/physiology ; Female ; *Host-Parasite Interactions ; Insect Vectors ; Larva ; Male ; Oocytes/microbiology ; Population Dynamics ; Reproduction ; Reverse Transcriptase Polymerase Chain Reaction ; Symbiosis/*physiology ; Trypanosoma/growth & development/isolation & purification ; Tsetse Flies/growth & development/microbiology/*parasitology/physiology ; Wolbachia/isolation & purification/pathogenicity/*physiology ; Zimbabwe ; }, abstract = {Symbiotic associations often enhance hosts' physiological capabilities, allowing them to expand into restricted terrains, thus leading to biological diversification. Stable maintenance of partners is essential for the overall biological system to succeed. The viviparous tsetse fly (Diptera: Glossinidae) offers an exceptional system to examine factors that influence the maintenance of multiple symbiotic organisms within a single eukaryotic host. This insect harbours three different symbionts representing diverse associations, coevolutionary histories and transmission modes. The enterics, obligate mutualist Wigglesworthia and beneficial Sodalis, are maternally transmitted to the intrauterine larvae, while parasitic Wolbachia infects the developing oocyte. In this study, the population dynamics of these three symbionts were examined through host development and during potentially disruptive events, including host immune challenge, the presence of third parties (such as African trypanosomes) and environmental perturbations (such as fluctuating humidity levels). While mutualistic partners exhibited well-regulated density profiles over different host developmental stages, parasitic Wolbachia infections varied in individual hosts. Host immune status and the presence of trypanosome infections did not impact the steady-state density levels observed for mutualistic microbes in either sex, while these factors resulted in an increase in Wolbachia density in males. Interestingly, perturbation of the maternal environment resulted in the deposition of progeny harbouring greater overall symbiont loads. The regulation of symbiont density, arising from coadaptive processes, may be an important mechanism driving inter-specific relations to ensure their competitive survival and to promote specialization of beneficial associations.}, } @article {pmid16618671, year = {2006}, author = {Fytrou, A and Schofield, PG and Kraaijeveld, AR and Hubbard, SF}, title = {Wolbachia infection suppresses both host defence and parasitoid counter-defence.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1588}, pages = {791-796}, pmid = {16618671}, issn = {0962-8452}, mesh = {Animals ; Body Size ; DNA Primers ; Drosophila/microbiology ; Female ; Gram-Negative Bacterial Infections/*physiopathology/*transmission ; Hymenoptera/microbiology ; Male ; Polymerase Chain Reaction ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Endosymbiotic bacteria in the genus Wolbachia have been linked to several types of reproductive parasitism, which enhance their own transmission, while their direct effects on the host vary from beneficial to neutral or detrimental. Here, we report negative effects of infection on immunity-related traits of Drosophila simulans and the parasitoid wasp Leptopilina heterotoma. Infected D. simulans showed a reduced ability to encapsulate parasitoid eggs, compared to a tetracycline-treated, bacterium-free line. Challenging the two lines with a fungal pathogen, Beauveria bassiana, on the other hand, revealed no differences in survival. Moreover, elimination of Wolbachia was beneficial for the parasitoid wasp, as eggs laid by uninfected females suffered significantly lower encapsulation rates. We discuss possible origins of these fitness costs and their implications for infection dynamics and the interactions between host species.}, } @article {pmid16616613, year = {2006}, author = {Heider, U and Blaxter, M and Hoerauf, A and Pfarr, KM}, title = {Differential display of genes expressed in the filarial nematode Litomosoides sigmodontis reveals a putative phosphate permease up-regulated after depletion of Wolbachia endobacteria.}, journal = {International journal of medical microbiology : IJMM}, volume = {296}, number = {4-5}, pages = {287-299}, doi = {10.1016/j.ijmm.2005.12.019}, pmid = {16616613}, issn = {1438-4221}, mesh = {Adaptation, Physiological/genetics ; Amino Acid Sequence ; Animals ; Anti-Bacterial Agents/pharmacology ; Filarioidea/*genetics/*microbiology/physiology ; Gene Expression Profiling ; *Gene Expression Regulation, Enzymologic ; Mice ; Molecular Sequence Data ; Phosphate Transport Proteins/biosynthesis/*genetics ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; *Symbiosis ; Tetracycline/pharmacology ; Up-Regulation ; Wolbachia/*drug effects/physiology ; }, abstract = {Mutualist symbiotic Wolbachia endobacteria are found in most filarial nematodes. Wolbachia are essential for embryogenesis and for larval development into adults, and thus represent a new target for anti-filarial drug development. Tetracycline antibiotics deplete Wolbachia in animal model filaria Litomosoides sigmodontis and Brugia pahangi, as well as in the human parasites Brugia malayi, Onchocerca volvulus and Wuchereria bancrofti. Very little is known about the molecular details of the symbiotic interaction between Wolbachia and filarial nematodes. Nematode genes that respond to anti-Wolbachia antibiotic treatment may play important roles in the symbiosis. Differential display PCR was used to detect several candidate genes that are up-regulated after 3, 6, 15, 30 and 36 days of tetracycline treatment. One of these genes, Ls-ppe-1, was similar to a family of phosphate permeases, and had putative orthologues in O. volvulus and B. malayi. Ls-ppe-1 steady-state mRNA levels were elevated by day 3-6 of treatment, and remained elevated through to 70 days post-treatment. In Caenorhabditis elegans, the knockdown of a homologous phosphate permease results in embryonic lethality, with the production of degenerating embryos, a phenotype also seen in filarial nematodes after depletion of Wolbachia with tetracycline. The potential role of Ls-ppe-1 in the nematode-bacterial symbiosis is discussed.}, } @article {pmid16615218, year = {2006}, author = {Duron, O and Fort, P and Weill, M}, title = {Hypervariable prophage WO sequences describe an unexpected high number of Wolbachia variants in the mosquito Culex pipiens.}, journal = {Proceedings. Biological sciences}, volume = {273}, number = {1585}, pages = {495-502}, pmid = {16615218}, issn = {0962-8452}, mesh = {Animals ; Culex/*microbiology ; Cytoplasm/virology ; DNA, Viral/chemistry/genetics ; Female ; Genetic Variation ; Male ; Open Reading Frames ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Prophages/*genetics ; Sequence Analysis, DNA ; Wolbachia/*genetics/*virology ; }, abstract = {Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI) resulting in abortive embryonic development. Among all the described host species, mosquitoes of the Culex pipiens complex display the highest variability of CI crossing types. Paradoxically, searches for polymorphism in Wolbachia infecting strains and field populations hitherto failed or produced very few markers. Here, we show that an abundant source of the long-sought polymorphism lies in WO prophage sequences present in multiple copies dispersed in the genome of Wolbachia infecting C. pipiens (wPip). We identified up to 66 different Wolbachia variants in C. pipiens strains and field populations and no occurrence of superinfection was observed. At least 49 different Wolbachia occurred in Southern Europe C. pipiens populations, and up to 10 different Wolbachia were even detected in a single population. This is in sharp contrast with North African and Cretan samples, which exhibited only six variants. The WO polymorphism appeared stable over time, and was exclusively transferred maternally. Interestingly, we found that the CI pattern previously described correlates with the variability of Gp15, a prophage protein similar to a bacterial virulence protein. WO prophage sequences thus represent variable markers that now open routes for approaching the molecular basis of CI, the host effects, the structure and dynamics of Wolbachia populations.}, } @article {pmid16610322, year = {2006}, author = {Duron, O and Labbé, P and Berticat, C and Rousset, F and Guillot, S and Raymond, M and Weill, M}, title = {High Wolbachia density correlates with cost of infection for insecticide resistant Culex pipiens mosquitoes.}, journal = {Evolution; international journal of organic evolution}, volume = {60}, number = {2}, pages = {303-314}, pmid = {16610322}, issn = {0014-3820}, mesh = {Animals ; Chlorpyrifos/*pharmacology ; Culex/*drug effects/genetics/*microbiology/physiology ; Female ; *Insecticide Resistance ; Insecticides/pharmacology ; Male ; Propoxur/*pharmacology ; Reproduction/drug effects/genetics/physiology ; Wolbachia/*physiology ; }, abstract = {In the mosquito Culex pipiens, insecticide resistance genes alter many life-history traits and incur a fitness cost. Resistance to organophosphate insecticides involves two loci, with each locus coding for a different mechanism of resistance (degradation vs. insensitivity to insecticides). The density of intracellular Wolbachia bacteria has been found to be higher in resistant mosquitoes, regardless of the mechanism involved. To discriminate between costs of resistance due to resistance genes from those associated with elevated Wolbachia densities, we compared strains of mosquito sharing the same genetic background but differing in their resistance alleles and Wolbachia infection status. Life-history traits measured included strength of insecticide resistance, larval mortality, adult female size, fecundity, predation avoidance, mating competition, and strength of cytoplasmic incompatibility (CI). We found that: (1) when Wolbachia are removed, insecticide resistance genes still affect some life-history traits; (2) Wolbachia are capable of modifying the cost of resistance; (3) the cost of Wolbachia infections increases with their density; (4) different interactions occurred depending on the resistance alleles involved; and (5) high densities of Wolbachia do not increase the strength of CI or maternal transmission efficiency relative to low Wolbachia densities. Insecticide resistance genes generated variation in the costs of Wolbachia infections and provided an interesting opportunity to study how these costs evolve, a process generally operating when Wolbachia colonizes a new host.}, } @article {pmid16599969, year = {2006}, author = {Narita, S and Nomura, M and Kato, Y and Fukatsu, T}, title = {Genetic structure of sibling butterfly species affected by Wolbachia infection sweep: evolutionary and biogeographical implications.}, journal = {Molecular ecology}, volume = {15}, number = {4}, pages = {1095-1108}, doi = {10.1111/j.1365-294X.2006.02857.x}, pmid = {16599969}, issn = {0962-1083}, mesh = {Animals ; Butterflies/classification/*genetics/*microbiology ; Genes, Mitochondrial ; *Geography ; Haplotypes ; Japan ; Models, Biological ; *Phylogeny ; *Wolbachia/genetics ; }, abstract = {It was recently recognized that in Japan, the common yellow butterfly, Eurema hecabe, consists of two sibling species, which have been unnamed yet and tentatively called yellow (Y) type and brown (B) type. We investigated the diversity of nuclear and mitochondrial genes in Japanese populations of Y type and B type of E. hecabe. The phylogeny based on nuclear genes agreed with the distinction between Y type and B type, which had been also supported by a wide array of biological data. However, the phylogeny based on mitochondrial genes did not reflect the distinction. PCR survey of Wolbachia revealed that B-type populations were all infected while Y-type populations contained both infected and uninfected individuals. A single genotype of Wolbachia, which was inferred to be a CI-inducing strain from their wsp gene sequence, was prevalent in these populations. Notably, the mitochondrial phylogeny was in perfect agreement with the pattern of Wolbachia infection, suggesting that the Wolbachia infection had affected the mitochondrial genetic structure of the host insects. Probably, the Wolbachia strain and the associated mitochondrial genomes have been occasionally introduced from B-type populations to Y-type populations through migration and subsequent interspecific hybridization, and CI-driven population sweep has been spreading the Wolbachia strain and the particular mitochondrial haplotypes, which originated from B-type populations, into Y-type populations. On the basis of these results together with the geological and biogeographical knowledge of the Japanese Archipelago, we proposed an evolutionary hypothesis on the invasion and spread of Wolbachia infection in B-type and Y-type of E. hecabe.}, } @article {pmid16598632, year = {2006}, author = {Sintupachee, S and Milne, JR and Poonchaisri, S and Baimai, V and Kittayapong, P}, title = {Closely related Wolbachia strains within the pumpkin arthropod community and the potential for horizontal transmission via the plant.}, journal = {Microbial ecology}, volume = {51}, number = {3}, pages = {294-301}, pmid = {16598632}, issn = {0095-3628}, mesh = {Animals ; Coleoptera/microbiology ; Cucurbita/*microbiology/parasitology ; Hemiptera/microbiology ; Insect Vectors/*microbiology ; Luffa/microbiology/parasitology ; Phylogeny ; Plant Diseases/*microbiology/parasitology ; Seasons ; Wolbachia/*classification/genetics/growth & development ; }, abstract = {Phylogenetic studies have implicated frequent horizontal transmission of Wolbachia among arthropod host lineages. However, the ecological routes for such lateral transfer are poorly known. We surveyed the species of two arthropod communities, one on pumpkin and the other on loofah plants, for Wolbachia, constructed wsp gene phylogenies of those Wolbachia strains found to infect community members, and established ecological links among infected members. Four taxonomically diverse insects in the pumpkin arthropod community contained very closely related Wolbachia wsp sequences (<1.5% divergence by Kimura-2-parameter distances). These insects, namely, the whitefly Bemisia tabaci, the planthopper Nisia nervosa, the flea beetle Phyllotreta sp., and the fleahopper Halticus minutus, were all collected from pumpkin leaves. They were ecologically linked through feeding on the same leaf substrate. Unlike other infected leaf insects, the whitefly population appeared to have a permanent breeding relationship with pumpkin plants, and high and stable, but not fixed, monthly Wolbachia infection rates. Our findings suggest potential roles for the plant in Wolbachia transmission and for whiteflies in being an infection source for other pumpkin leaf-feeding insects.}, } @article {pmid16597203, year = {2006}, author = {Hoerauf, A}, title = {New strategies to combat filariasis.}, journal = {Expert review of anti-infective therapy}, volume = {4}, number = {2}, pages = {211-222}, doi = {10.1586/14787210.4.2.211}, pmid = {16597203}, issn = {1744-8336}, mesh = {Animals ; Doxycycline/*administration & dosage/therapeutic use ; Drug Delivery Systems/methods/*trends ; Filariasis/*drug therapy/microbiology ; Filaricides/*administration & dosage/therapeutic use ; Humans ; }, abstract = {Two of the major filarial infections, lymphatic filariasis (LF) and onchocerciasis, affect 150 million people, while 1 billion living in endemic areas are at risk of infection. Public health programs to control these infections have successfully existed for years and have evolved from activities driven by the WHO into global programs with public-private partnerships. Currently, these programs use yearly mass application of drugs that mainly kill the larval stages (the microfilariae), with the aim of preventing uptake by the transmitting insect vectors and thus, to block transmission and reduce the infections to such levels that in 15-30 years from now, they will no longer pose a public health problem. While the programs have been very successful in general, there are drawbacks such as coverage being too low within the population, reappearance of infection by migration of infected people into controlled areas, targeting of a stage (the microfilaria) that does not induce pathology in LF and thus lowers compliance, and the potential development of drug resistance, first indications of which have been clearly observed in onchocerciasis. In addition, even without drawbacks, program scopes are not the eradication of filarial infections, which is, however, an ultimate goal of control activities. There is therefore an unequivocal call for the development of higher efficient, complementary chemotherapeutical approaches that lead to a long-lasting reduction of the pathology-inducing worm stages; that is, microfilariae in onchocerciasis and adult worms in LF, or to a macrofilaricidal effect. The recent discovery that depletion of Wolbachia endosymbionts by tetracycline antibiotics leads to long-lasting sterility of adult female worms in onchocerciasis and a macrofilaricidal effect in LF fulfils these requirements. Successful regimens have already been published and agreed upon for use by expert panels. While these regimens are still too long for mass application, the antiwolbachial chemotherapy can currently be applied in the form of a suitable doxycycline regimen for 6 weeks for the treatment of individuals, and exploited in the future for the development of new drugs suitable for mass application. In addition, first data suggest that Wolbachia may also be major mediators of lymphangiogenesis and that their depletion is associated with reduction of lymph vessel-specific vascular endothelial growth factors and reduced lymph vessel size.}, } @article {pmid16596351, year = {2006}, author = {Reeves, WK and Dowling, AP and Dasch, GA}, title = {Rickettsial agents from parasitic dermanyssoidea (Acari: Mesostigmata).}, journal = {Experimental & applied acarology}, volume = {38}, number = {2-3}, pages = {181-188}, pmid = {16596351}, issn = {0168-8162}, mesh = {Anaplasma/genetics/isolation & purification ; Animals ; Bartonella/genetics/isolation & purification ; Base Sequence ; DNA, Bacterial/chemistry/genetics ; Gram-Negative Bacteria/genetics/*isolation & purification ; Mites/*microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/chemistry/genetics ; Rickettsia/genetics/isolation & purification ; Sequence Analysis, DNA ; Spiroplasma/genetics/isolation & purification ; Wolbachia/genetics/isolation & purification ; }, abstract = {Mites are often overlooked as vectors of pathogens, but have been shown to harbor and transmit rickettsial agents such as Rickettsia akari and Orientia tsutsugamushi. We screened DNA extracts from 27 mites representing 25 species of dermanyssoids for rickettsial agents such as Anaplasma, Bartonella, Rickettsia, and Wolbachia by PCR amplification and sequencing. DNA from Anaplasma spp., a novel Bartonella sp., Spiroplasma sp., Wolbachia sp., and an unclassified Rickettsiales were detected in mites. These could represent mite-borne bacterial agents, bacterial DNA from blood meals, or novel endosymbionts of mites.}, } @article {pmid16575725, year = {2006}, author = {Lammie, PJ}, title = {The promise of wolbachia-targeted chemotherapy as a public health intervention for lymphatic filariasis and onchocerciasis.}, journal = {Clinical infectious diseases : an official publication of the Infectious Diseases Society of America}, volume = {42}, number = {8}, pages = {1090-1092}, doi = {10.1086/501361}, pmid = {16575725}, issn = {1537-6591}, mesh = {Anthelmintics/therapeutic use ; Filariasis/*complications ; Gram-Negative Bacterial Infections/*drug therapy ; Humans ; Onchocerciasis/*complications ; *Wolbachia ; }, } @article {pmid16575724, year = {2006}, author = {Turner, JD and Mand, S and Debrah, AY and Muehlfeld, J and Pfarr, K and McGarry, HF and Adjei, O and Taylor, MJ and Hoerauf, A}, title = {A randomized, double-blind clinical trial of a 3-week course of doxycycline plus albendazole and ivermectin for the treatment of Wuchereria bancrofti infection.}, journal = {Clinical infectious diseases : an official publication of the Infectious Diseases Society of America}, volume = {42}, number = {8}, pages = {1081-1089}, doi = {10.1086/501351}, pmid = {16575724}, issn = {1537-6591}, mesh = {Adolescent ; Adult ; Aged ; Albendazole/*therapeutic use ; Animals ; Anthelmintics/therapeutic use ; Double-Blind Method ; Doxycycline/*therapeutic use ; Drug Therapy, Combination ; Filariasis/*drug therapy ; Ghana ; Humans ; Ivermectin/*therapeutic use ; Middle Aged ; Patient Dropouts ; *Wuchereria bancrofti ; }, abstract = {BACKGROUND: Eight- and 6-week courses of doxycycline are superior to standard treatment of bancroftian filariasis. Standard treatment (albendazole plus ivermectin) is associated with adverse reactions. We assessed whether a shorter (i.e, 3-week) course of doxycycline with standard treatment would show superior efficacy to standard treatment alone and reduce the incidence of adverse reactions.

METHODS: A total of 44 adults from Ghana were recruited in January 2003: 20 received doxycycline (200 mg/day) for 3 weeks, and 24 received matching placebo. Participants received albendazole (400 mg) and ivermectin (150 microg/kg) at month 4, and adverse reactions were assessed 48 h later. Treatment efficacy was evaluated at months 4, 12, and 24.

RESULTS: The microfilariae level was significantly reduced after receipt of doxycycline treatment at months 4 (P = .017), 12 (P = .001), and 24 (P = .005). The microfilariae level was only significantly reduced at month 12 in the placebo group (P = .041). At all follow-up points, the microfilariae level was significantly lower in the doxycycline group. Adverse reactions to standard antifilarial treatment were similar in frequency between the doxycycline group (in 7 of 11 subjects) and the placebo group (in 13 of 17 subjects). Moderate reactions only occurred in the placebo group (in 3 of 17 subjects). Severity of adverse reaction was associated with microfilaremia (P = .037), Wolbachia bacteria in plasma (P = .048), and proinflammatory cytokines in plasma (P = .019). Adult parasite viability was not significantly different between doxycycline and placebo groups at months 12 or 24.

CONCLUSIONS: Treatment with doxycycline for 3 weeks is more effective in inducing a long-term amicrofilaremia than is standard treatment alone, but it is ineffective at inducing curative effects. Inflammatory reactions to antifilarial treatment are associated with levels of microfilariae and Wolbachia endosymbionts released into plasma.}, } @article {pmid16568634, year = {2006}, author = {Koukou, K and Pavlikaki, H and Kilias, G and Werren, JH and Bourtzis, K and Alahiotis, SN}, title = {Influence of antibiotic treatment and Wolbachia curing on sexual isolation among Drosophila melanogaster cage populations.}, journal = {Evolution; international journal of organic evolution}, volume = {60}, number = {1}, pages = {87-96}, pmid = {16568634}, issn = {0014-3820}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Crosses, Genetic ; Drosophila melanogaster/drug effects/*microbiology/*physiology ; Female ; Genetic Speciation ; Male ; Reproduction ; Sexual Behavior, Animal ; Tetracycline/*pharmacology ; Wolbachia/*drug effects/physiology ; }, abstract = {Speciation depends on the establishment of reproductive isolation between populations of the same species. Whether assortative mating evolves as a by-product of adaptation is a major question relevant to the origin of species by reproductive isolation. The long-term selection populations used here were originally established 30 years ago from a single cage population (originating from a maternal one) and subsequently subjected to divergent selection for tolerance of toxins in food (heavy metals versus ethanol) to investigate this question. Those populations now differ in sexual isolation and Wolbachia infection status. Wolbachia are common and widespread bacteria infecting arthropods and nematodes. Attention has recently focused on their potential role in insect speciation, due to post-mating sperm-egg incompatibilities induced by the bacteria. In this paper we examine the potential effect of Wolbachia on the level of sexual isolation. By antibiotic curing, we show that removal of Wolbachia decreases levels of mate discrimination (sexual isolation index) between populations by about 50%. Backcrossing experiments confirm that this effect is due to infection status rather than to genetic changes in the populations resulting from antibiotic treatment. Antibiotic treatment has no effect on mate discrimination level between uninfected populations. Our findings suggest that the presence of Wolbachia (or another undetected bacterial associate) act as an additive factor contributing to the level of pre-mating isolation between these Drosophila melanogaster populations. Given the ubiquity of bacterial associates of insects, such effects could be relevant to some speciation events.}, } @article {pmid16563429, year = {2006}, author = {Zeh, JA and Zeh, DW}, title = {Male-killing Wolbachia in a live-bearing arthropod: brood abortion as a constraint on the spread of a selfish microbe.}, journal = {Journal of invertebrate pathology}, volume = {92}, number = {1}, pages = {33-38}, doi = {10.1016/j.jip.2006.02.004}, pmid = {16563429}, issn = {0022-2011}, mesh = {Animals ; Arthropods/*parasitology ; Female ; Host-Parasite Interactions ; Male ; *Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Maternally inherited, cellular endosymbionts can enhance their fitness by biasing host sex ratio in favor of females. Male killing (MK), an extreme form of sex-ratio manipulation, is selectively advantageous, if the death of males results in increased microbe transmission through female siblings. In live-bearing hosts, females typically produce more embryos than are brought to term, and reproductive compensation through maternal resource reallocation from dead male embryos to female siblings provides a direct, physiological mechanism that could increase the number of daughters born to infected females, thereby promoting MK endosymbiont spread. In this study, a Wolbachia-infected line and an uninfected line of the viviparous pseudoscorpion, Cordylochernes scorpioides were genetically homogenized for nuclear DNA by repeated backcrossing of the infected line with the uninfected, laboratory population. Photomicroscopy of early-stage embryos demonstrated that female C. scorpioides invariably produced an excess of embryos, with Wolbachia-infected females producing as many early-stage embryos as uninfected female controls. However, Wolbachia-infected females that successfully carried broods to term gave birth to significantly fewer offspring, indicating that the extreme female bias characteristic of their broods results from the killing rather than the feminization of male embryos. Infected females that carried broods to term gave birth to significantly larger nymphs and did produce 10% more female offspring than uninfected females. However, the slight transmission advantage that the MK Wolbachia accrued from this reproductive compensation appears to be heavily outweighed by the high rate of spontaneous brood abortion suffered by infected females.}, } @article {pmid16563157, year = {2006}, author = {Townson, S and Tagboto, S and McGarry, HF and Egerton, GL and Taylor, MJ}, title = {Onchocerca parasites and Wolbachia endosymbionts: evaluation of a spectrum of antibiotic types for activity against Onchocerca gutturosa in vitro.}, journal = {Filaria journal}, volume = {5}, number = {}, pages = {4}, pmid = {16563157}, issn = {1475-2883}, abstract = {BACKGROUND: The filarial parasites of major importance in humans contain the symbiotic bacterium Wolbachia and recent studies have shown that targeting of these bacteria with antibiotics results in a reduction in worm viability, development, embryogenesis, and survival. Doxycycline has been effective in human trials, but there is a need to develop drugs that can be given for shorter periods and to pregnant women and children. The World Health Organisation-approved assay to screen for anti-filarial activity in vitro uses male Onchocerca gutturosa, with effects being determined by worm motility and viability as measured by reduction of MTT to MTT formazan. Here we have used this system to screen antibiotics for anti-filarial activity. In addition we have determined the contribution of Wolbachia depletion to the MTT reduction assay.

METHODS: Adult male O. gutturosa were cultured on a monkey kidney cell (LLCMK 2) feeder layer in 24-well plates with antibiotics and antibiotic combinations (6 to 10 worms per group). The macrofilaricide CGP 6140 (Amocarzine) was used as a positive control. Worm viability was assessed by two methods, (i) motility levels and (ii) MTT/formazan colorimetry. Worm motility was scored on a scale of 0 (immotile) to 10 (maximum) every 5 days up to 40 days. On day 40 worm viability was evaluated by MTT/formazan colorimetry, and results were expressed as a mean percentage reduction compared with untreated control values at day 40. To determine the contribution of Wolbachia to the MTT assay, the MTT formazan formation of an insect cell-line (C6/36) with or without insect Wolbachia infection and treated or untreated with tetracycline was compared.

RESULTS: Antibiotics with known anti-Wolbachia activity were efficacious in this system. Rifampicin (5 x 10(-5) M) was the most effective anti-mycobacterial agent; clofazimine (1.25 x 10(-5) M and 3.13 x 10(-6) M) produced a gradual reduction in motility and by 40 days had reduced worm viability. The other anti-mycobacterial drugs tested had limited or no activity. Doxycycline (5 x 10(-5) M) was filaricidal, but minocycline was more effective and at a lower concentration (5 x 10(-5) M and 1.25 x 10(-5) M). Inactive compounds included erythromycin, oxytetracycline, trimethoprim and sulphamethoxazole. The MTT assay on the insect cell-line showed that Wolbachia made a significant contribution to the metabolic activity within the cells, which could be reduced when they were exposed to tetracycline.

CONCLUSION: The O. gutturosa adult male screen for anti-filarial drug activity is also valid for the screening of antibiotics for anti-Wolbachia activity. In agreement with previous findings, rifampicin and doxycycline were effective; however, the most active antibiotic was minocycline. Wolbachia contributed to the formation of MTT formazan in the MTT assay of viability and is therefore not exclusively a measure of worm viability and indicates that Wolbachia contributes directly to the metabolic activity of the nematode.}, } @article {pmid16552075, year = {2006}, author = {Gillette-Ferguson, I and Hise, AG and Sun, Y and Diaconu, E and McGarry, HF and Taylor, MJ and Pearlman, E}, title = {Wolbachia- and Onchocerca volvulus-induced keratitis (river blindness) is dependent on myeloid differentiation factor 88.}, journal = {Infection and immunity}, volume = {74}, number = {4}, pages = {2442-2445}, pmid = {16552075}, issn = {0019-9567}, support = {P30 EY011373/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; K08 AI054652/AI/NIAID NIH HHS/United States ; R01 EY010320/EY/NEI NIH HHS/United States ; T32 AI007024/AI/NIAID NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; //Wellcome Trust/United Kingdom ; AI-07024/AI/NIAID NIH HHS/United States ; }, mesh = {Adaptor Proteins, Signal Transducing/biosynthesis/deficiency/genetics/*physiology ; Animals ; Disease Models, Animal ; Eye Infections, Bacterial/genetics/*immunology ; Keratitis/genetics/*immunology ; Mice ; Mice, Knockout ; Myeloid Differentiation Factor 88 ; Neutrophils/immunology ; Onchocerca volvulus/*immunology ; Onchocerciasis, Ocular/genetics/*immunology ; Signal Transduction/genetics/immunology ; Toll-Like Receptors/genetics/physiology ; Wolbachia/*immunology/pathogenicity ; }, abstract = {Endosymbiotic Wolbachia bacteria that infect the filarial nematode Onchocerca volvulus were previously found to have an essential role in the pathogenesis of river blindness. The current study demonstrates that corneal inflammation induced by Wolbachia or O. volvulus antigens containing Wolbachia is completely dependent on expression of myeloid differentiation factor 88.}, } @article {pmid16547041, year = {2006}, author = {Frutos, R and Viari, A and Ferraz, C and Morgat, A and Eychenié, S and Kandassamy, Y and Chantal, I and Bensaid, A and Coissac, E and Vachiery, N and Demaille, J and Martinez, D}, title = {Comparative genomic analysis of three strains of Ehrlichia ruminantium reveals an active process of genome size plasticity.}, journal = {Journal of bacteriology}, volume = {188}, number = {7}, pages = {2533-2542}, pmid = {16547041}, issn = {0021-9193}, mesh = {Conserved Sequence ; Ehrlichia ruminantium/*classification/*genetics ; *Evolution, Molecular ; Gene Order ; Genetic Variation/*genetics ; *Genome, Bacterial ; Molecular Sequence Data ; Mutagenesis/*genetics ; Phenotype ; Species Specificity ; Tandem Repeat Sequences/genetics ; }, abstract = {Ehrlichia ruminantium is the causative agent of heartwater, a major tick-borne disease of livestock in Africa that has been introduced in the Caribbean and is threatening to emerge and spread on the American mainland. We sequenced the complete genomes of two strains of E. ruminantium of differing phenotypes, strains Gardel (Erga; 1,499,920 bp), from the island of Guadeloupe, and Welgevonden (Erwe; 1,512,977 bp), originating in South Africa and maintained in Guadeloupe in a different cell environment. Comparative genomic analysis of these two strains was performed with the recently published parent strain of Erwe (Erwo) and other Rickettsiales (Anaplasma, Wolbachia, and Rickettsia spp.). Gene order is highly conserved between the E. ruminantium strains and with A. marginale. In contrast, there is very little conservation of gene order with members of the Rickettsiaceae. However, gene order may be locally conserved, as illustrated by the tuf operons. Eighteen truncated protein-encoding sequences (CDSs) differentiate Erga from Erwe/Erwo, whereas four other truncated CDSs differentiate Erwe from Erwo. Moreover, E. ruminantium displays the lowest coding ratio observed among bacteria due to unusually long intergenic regions. This is related to an active process of genome expansion/contraction targeted at tandem repeats in noncoding regions and based on the addition or removal of ca. 150-bp tandem units. This process seems to be specific to E. ruminantium and is not observed in the other Rickettsiales.}, } @article {pmid16542426, year = {2006}, author = {Rao, RU and Klei, TR}, title = {Cytokine profiles of filarial granulomas in jirds infected with Brugia pahangi.}, journal = {Filaria journal}, volume = {5}, number = {}, pages = {3}, pmid = {16542426}, issn = {1475-2883}, abstract = {BACKGROUND: A granulomatous inflammatory response develops in jirds infected subcutaneously or intraperitoneally with filarial nematodes namely Brugia pahangi and B. malayi. Previous studies by light and electron microscopy have shown cellular inflammatory responses in and around these granulomas. Furthermore, the cellular inflammatory responses of granulomas found in the lymphatics and peritoneal cavity appear to be similar. The purpose of this study was to determine the cytokine profiles of granulomas in the peritoneal cavity of B. pahangi-infected jirds and to determine whether the granulomas release any proinflammatory cytokines ex vivo.

METHODS: A semiautomated quantitative polymerase chain reaction (Q-PCR) was performed on cDNA prepared from the granulomas of infected jirds to study the species-specific mRNA expression of IL-2, IL-4, IFN-gamma, IL-5, and IL-10. Genomic DNA was extracted from the granulomas, and parasite DNA was detected by Q-PCR by amplifying the HhaI repeat sequence. The levels of the inflammation-causing cytokines IL-6 and TNFalpha that were secreted by the granulomas were measured by cell-based assays.

RESULTS: Florid granulomas showed higher levels of IFN-gamma than other cytokines, linking this Th1 cytokine to the granulomatous inflammation that develops in jirds and humans. IL-4 expression was much lower than that of IFN-gamma but higher than that of IL-10. A low level of IL-5 mRNA expression was detectable in all granulomas as was the level of IL-2 expression. The levels of the inflammatory cytokines IL-6 and TNFalpha, secreted by intact granulomas, spontaneously increased by 48 h after culture. Parasite antigen stimulation and subsequent release of IL-6 and TNFalpha by the granulomas indicated a moderate increase in the levels of these two cytokines. The amplification of the Brugia HhaI repeat DNA and Wolbachia 16S rDNA indicated worm components and bacterial components in the granulomatous tissue.

CONCLUSION: Granuloma development in filarial infections is a complex process involving cellular reactions responding to parasite/bacteria and their components. The interactions between worm-derived granulomas and their hosts are dynamic and multifaceted. The data collected thus far suggest that the expression profiles of many of the measured cytokines in the lymphoid tissues of Brugia-infected jirds are different from those of the cytokines in granulomas. Moreover, granulomas have the ability to secrete the inflammatory cytokines IL-6 and TNFalpha.}, } @article {pmid16528802, year = {2006}, author = {Thomson, N and Crossman, L and Bentley, S}, title = {Bacterial home from home.}, journal = {Nature reviews. Microbiology}, volume = {4}, number = {3}, pages = {168-170}, doi = {10.1038/nrmicro1367}, pmid = {16528802}, issn = {1740-1526}, mesh = {Animals ; Brugia malayi/metabolism/microbiology ; Enterobacteriaceae/*genetics/metabolism ; Evolution, Molecular ; Gammaproteobacteria/*genetics/metabolism ; *Genome, Bacterial ; Insecta/microbiology ; *Symbiosis ; Trypanosoma brucei brucei/microbiology ; Tsetse Flies/microbiology/physiology ; Wolbachia/*genetics/metabolism ; }, } @article {pmid16495252, year = {2006}, author = {Makepeace, BL and Rodgers, L and Trees, AJ}, title = {Rate of elimination of Wolbachia pipientis by doxycycline in vitro increases following drug withdrawal.}, journal = {Antimicrobial agents and chemotherapy}, volume = {50}, number = {3}, pages = {922-927}, pmid = {16495252}, issn = {0066-4804}, mesh = {Aedes/cytology ; Animals ; Anti-Bacterial Agents/*pharmacology ; Cell Line ; Doxycycline/*pharmacology ; Polymerase Chain Reaction ; RNA, Ribosomal/genetics ; RNA, Ribosomal, 16S/genetics ; *Substance Withdrawal Syndrome ; Time Factors ; Wolbachia/*drug effects/genetics/*physiology ; }, abstract = {Wolbachia pipientis is an obligate intracellular bacterium within the family Anaplasmataceae that infects many terrestrial arthropods and arthropod-transmitted nematodes (filariae). Several filarial species are major human pathogens, and antibiotics with activity against Wolbachia offer a promising new therapeutic approach, since the adult worms are relatively refractory to conventional anthelmintics but depend on Wolbachia for reproduction and viability. In a natural filarial parasite of cattle, Onchocerca ochengi, intermittent chemotherapy is adulticidal whereas the equivalent dose administered as a continuous treatment is not. To investigate this further and to aid the design of efficacious regimens for human therapy, we used Wolbachia-infected Aedes albopictus mosquito cells in vitro. Here, we describe for the first time the accelerated depletion of bacteria after antibiotic withdrawal relative to the rate of elimination in the continuous presence of the drug. Mosquito cells were incubated with doxycycline while changes in 16S (bacterial) and 18S (host) rRNA and rRNA genes were determined by quantitative PCR assays. In cultures treated for 7 or 14 days followed by 7 days of drug withdrawal, the Wolbachia-to-Aedes rRNA ratio declined by approximately 6 log, whereas immediately after 14 or 21 days of continuous treatment, the reduction was only approximately 4 log (P < 0.05). However, low levels of 16S rRNA remained after 21 days of treatment, irrespective of whether doxycycline was withdrawn. Application of similar methodology to related intracellular bacteria may reveal that this posttreatment effect is not restricted to Wolbachia and could have wider implications for the design of intermittent regimens for antibiotic chemotherapy.}, } @article {pmid16493396, year = {2004}, author = {Mak, JW}, title = {Antifilarial compounds in the treatment and control of lymphatic filariasis.}, journal = {Tropical biomedicine}, volume = {21}, number = {2}, pages = {27-38}, pmid = {16493396}, issn = {0127-5720}, abstract = {Diethylcarbamazine citrate (DEC) has been used for treatment and control of lymphatic filariasis since the 1950s. Although this remarkable drug is still useful and modified strategies in its usage have been developed, a number of newer antifilarial compounds are now available. Numerous field trials evaluating their efficacy in the control of lymphatic filariasis have been conducted. In particular, ivermectin (IVM), albendazole (ALB), and DEC have been tested singly and in combinations and the results of such field studies should be evaluated. While most of the studies were based on efficacy in the clearance of microfilaraemia, a few clinical trials evaluated the adulticidal activity of these compounds. Some antibiotics are effective in killing Wolbachia bacteria symbionts of filarial worms, but their role in the chemotherapy of lymphatic filariasis is still undefined. This review of randomised controlled field studies and randomised controlled clinical trials with these compounds will summarise the findings and give recommendations on their appropriate use for the control and treatment of lymphatic filariasis.}, } @article {pmid16489228, year = {2006}, author = {Clark, ME and Heath, BD and Anderson, CL and Karr, TL}, title = {Induced paternal effects mimic cytoplasmic incompatibility in Drosophila.}, journal = {Genetics}, volume = {173}, number = {2}, pages = {727-734}, pmid = {16489228}, issn = {0016-6731}, mesh = {Animals ; Animals, Genetically Modified ; Cytoplasm/microbiology ; Drosophila/*genetics/*microbiology/physiology ; Drosophila Proteins/genetics ; Drosophila melanogaster/genetics/microbiology/physiology ; Female ; Gene Expression ; Genes, Insect ; Male ; Membrane Proteins/genetics ; Models, Biological ; Myosin Heavy Chains/genetics ; Ovum/microbiology ; Phenotype ; Reproduction ; Spermatogenesis ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is an intracellular microbe found in a wide diversity of arthropod and filarial nematode hosts. In arthropods these common bacteria are reproductive parasites that manipulate central elements of their host's reproduction to increase their own maternal transmission in one of several ways. Cytoplasmic incompatibility (CI) is one such manipulation where sperm are somehow modified in infected males and this modification must be rescued by the presence of the same bacterial strain in the egg for normal development to proceed. The molecular mechanisms involved in the expression of CI are unknown. Here we show that Wolbachia infection results in increased mRNA and protein expression of the Drosophila simulans nonmuscle myosin II gene zipper. Induced overexpression of zipper in Wolbachia-free transgenic D. melanogaster males results in paternal-effect lethality that mimics the fertilization defects associated with CI. Likewise, overexpression of the tumor suppressor gene, lethal giant larvae [l(2)gl], results in egg lethality and a CI phenotype. Stoichiometric levels of zipper and l(2)gl are required for proper segregation of cellular determinants during neuroblast stem cell division. Taken together these results form the basis of a working hypothesis whereby Wolbachia induces paternal effects in sperm by manipulating the expression of key regulators of cytoskeletal activity during spermatogenesis.}, } @article {pmid16472187, year = {2006}, author = {Pfarr, KM and Hoerauf, AM}, title = {Antibiotics which target the Wolbachia endosymbionts of filarial parasites: a new strategy for control of filariasis and amelioration of pathology.}, journal = {Mini reviews in medicinal chemistry}, volume = {6}, number = {2}, pages = {203-210}, doi = {10.2174/138955706775475984}, pmid = {16472187}, issn = {1389-5575}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Filariasis/*drug therapy ; Filarioidea/*microbiology/physiology ; Humans ; *Symbiosis ; Wolbachia/*drug effects ; }, abstract = {Wolbachia endosymbionts of filariae are targets for the development of new antifilarial chemotherapy. Doxycycline to deplete Wolbachia from the worm has demonstrated the feasibility of this strategy and has provided a new chemotherapeutic tool. Recent research shows that depleting Wolbachia will also lessen pathology, and lessen adverse reactions to traditional antifilarial drugs.}, } @article {pmid16469072, year = {2006}, author = {Bouwma, AM and Ahrens, ME and DeHeer, CJ and DeWayne Shoemaker, D}, title = {Distribution and prevalence of Wolbachia in introduced populations of the fire ant Solenopsis invicta.}, journal = {Insect molecular biology}, volume = {15}, number = {1}, pages = {89-93}, doi = {10.1111/j.1365-2583.2006.00614.x}, pmid = {16469072}, issn = {0962-1075}, mesh = {Animals ; Ants/genetics/*microbiology ; Female ; Male ; Southeastern United States ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Wolbachia are intracellular bacteria that induce phenotypic effects in many arthropod hosts to enhance their own transmission within host populations. Wolbachia commonly infect the Red Imported Fire Ant, Solenopsis invicta, in native South American populations. A previous study failed to detect Wolbachia in fire ants from the introduced range in the USA. We conducted an extensive study of individuals collected from 1157 nests from 10 widespread geographical populations in the USA. Wolbachia were detected in ants from two nearby populations in southern Mississippi, with different variants (wsp gene sequences) infecting ants from colonies of the multiple-queen (polygyne) vs. single-queen (monogyne) social forms. The parsimonious explanation for the presence of Wolbachia in introduced S. invicta is that there have been one or more recent introductions of Wolbachia-infected fire ants into the southern USA.}, } @article {pmid16457735, year = {2006}, author = {Debrah, AY and Mand, S and Marfo-Debrekyei, Y and Larbi, J and Adjei, O and Hoerauf, A}, title = {Assessment of microfilarial loads in the skin of onchocerciasis patients after treatment with different regimens of doxycycline plus ivermectin.}, journal = {Filaria journal}, volume = {5}, number = {}, pages = {1}, pmid = {16457735}, issn = {1475-2883}, abstract = {BACKGROUND: Infection with the filarial nematode Onchocerca volvulus can lead to severe dermatitis, visual impairment, and ultimately blindness. Since the currently used drug, ivermectin does not have macrofilaricidal or strong permanent sterilising effects on the adult worm, more effective drugs are needed to complement the use of ivermectin alone. Wolbachia endosymbiotic bacteria in filariae have emerged as a new target for treatment with antibiotics which can lead to long -term sterilization of the adult female filariae.

METHODS: In the Central Region of Ghana, 60 patients were recruited, allocated into four groups and treated with 200 mg doxycycline per day for 2 weeks, 4 weeks, 6 weeks respectively. Untreated patients served as controls. Some of the treated patients and the untreated controls were given 150 microg/kg ivermectin 8 months after the start of doxycycline treatment.

RESULTS: A follow up study 18 months post treatment showed that when using doxycycline alone there was a significant reduction of microfilarial (mf) loads in patients treated for either 4 or 6 weeks. However, there was no significant difference between the untreated controls and those given the 2 weeks regimen. Although no significant difference was demonstrated between the 4 and 6 weeks regimens, there was a trend observed, in that, microfilarial reduction appeared to have been greater following the 6 weeks regimen. Twelve months after ivermectin (i.e. 20 months after doxycycline) treatment, 8 out of 11 ivermectin-alone treated patients were mf-positive. In contrast, 1 out of the 7 patients treated for 4 weeks with doxycycline and none of the 4 patients treated for 6 weeks doxycycline (who were available for re-examination) were mf-positive after the combined treatment of doxycycline plus ivermectin treatment.

CONCLUSION: Treatment of onchocerciasis with doxycycline for 4 weeks is effective. Nonetheless, mf reduction appeared to be greater in the 6 weeks regimen. It is recommended that until further studies are carried out i.e. 4 weeks treatment with doxycycline is proven equivalent to the 6 weeks, selected groups of onchocerciasis patients should be treated for 6 weeks with doxycycline. As discussed earlier, this treatment should be accompanied by two doses of ivermectin.}, } @article {pmid16448417, year = {2006}, author = {Goodacre, SL and Martin, OY and Thomas, CF and Hewitt, GM}, title = {Wolbachia and other endosymbiont infections in spiders.}, journal = {Molecular ecology}, volume = {15}, number = {2}, pages = {517-527}, doi = {10.1111/j.1365-294X.2005.02802.x}, pmid = {16448417}, issn = {0962-1083}, mesh = {Animal Diseases/microbiology ; Animals ; Female ; Male ; Molecular Sequence Data ; *Phylogeny ; Rickettsia/genetics ; Spiders/*microbiology ; Spiroplasma/genetics ; Symbiosis ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Maternally inherited endosymbiotic bacteria, such as Wolbachia, Rickettsia and Spiroplasma, have been shown to have wide-ranging effects on the reproduction of their hosts. We present data on the presence of each of these sorts of bacteria in spiders, a group for which there are currently few data, but where such infections could explain many observed reproductive characteristics, such as sex ratio skew. The Wolbachia and Spiroplasma variants that we find in spiders belong to the same clades previously found to infect other arthropods, but many of the rickettsias belong to two, novel, hitherto spider-specific bacterial lineages. We find evidence for coexistence of different bacterial types within species, and in some cases, within individuals. We suggest that spiders present a useful opportunity for studying the effect of these sorts of bacteria on the evolution of host traits, such as those that are under sexual selection.}, } @article {pmid16421757, year = {2006}, author = {Amat, I and Castelo, M and Desouhant, E and Bernstein, C}, title = {The influence of temperature and host availability on the host exploitation strategies of sexual and asexual parasitic wasps of the same species.}, journal = {Oecologia}, volume = {148}, number = {1}, pages = {153-161}, pmid = {16421757}, issn = {0029-8549}, mesh = {Animals ; Behavior, Animal/*physiology ; Female ; Host-Parasite Interactions ; Lepidoptera/*parasitology ; Oviposition ; Reproduction/physiology ; Reproduction, Asexual/physiology ; *Temperature ; Wasps/*physiology ; }, abstract = {In the hymenopteran parasitoid Venturia canescens, asexual (obligate thelytoky not induced by Wolbachia bacteria) and sexual (arrhenotokous) wasps coexist in field conditions despite the demographic cost incurred due to the production of males by sexual females. Arrhenotoky predominates in field conditions, whereas populations in indoor conditions (mills, granaries) are exclusively thelytokous. These differences in the relative abundance of the two modes of reproduction between environments suggest that the individuals of each reproductive mode may have developed strategies adapted to the conditions prevailing in each kind of habitat. The two environments contrast in temperature variability and in the spatial heterogeneity of host availability. In this study, we considered the combined effect of temperature and host availability on host patch exploitation by thelytokous and arrhenotokous V. canescens. As expected, arrhenotokous females were more sensitive to temperature changes. If the temperature decreased before foraging, they remained longer and exploited patches more thoroughly. This is consistent with the expected behaviour of parasitoids in response to signs of unfavourable conditions that entail increasing risk of time limitation or a reduced probability of attaining further patches. Both arrhenotokous and thelytokous females increased patch exploitation with host availability. However, unexpectedly, we found no difference in the way the two types of wasp responded to differences in host availability. Differences in the strategies adopted under different environmental conditions may indicate divergence of niche-specific life history traits between the two modes of reproduction. Niche displacement may partly account for the coexistence of these two modes of reproduction at a geographical scale.}, } @article {pmid16406333, year = {2006}, author = {Fenn, K and Blaxter, M}, title = {Wolbachia genomes: revealing the biology of parasitism and mutualism.}, journal = {Trends in parasitology}, volume = {22}, number = {2}, pages = {60-65}, doi = {10.1016/j.pt.2005.12.012}, pmid = {16406333}, issn = {1471-4922}, mesh = {Animals ; Arthropods/*microbiology ; Eukaryotic Cells/microbiology ; Filarioidea/*microbiology ; *Genome, Bacterial/genetics ; Phylogeny ; Rickettsiaceae/classification/genetics ; Symbiosis/*genetics ; Wolbachia/classification/*genetics/physiology ; }, abstract = {Wolbachia bacteria are endosymbiotic partners of many animal species, in which they behave as either parasites (in arthropod hosts) or mutualists (in nematode hosts). What biochemistry and biology underpin these diverse lifestyles? The recent complete sequencing of genomes from Wolbachia that infect the arthropod Drosophila melanogaster and the nematode Brugia malayi, together with the partial genome sequencing of three Wolbachia strains found in drosophilids, enables this question to begin to be addressed. Parasitic arthropod Wolbachia are characterized by the presence of phages that carry ankyrin-repeat proteins; these proteins might be exported to the host cell to manipulate reproduction. In nematode Wolbachia, which lack these phages, several biochemical pathways can deliver essential metabolites to the nematode hosts. Nematode Wolbachia might also have a role in modulating the mammalian host immune system but the sequenced Wolbachia genomes lack the genes to synthesize lipopolysaccharide, raising questions about the nature of the inducing molecule. The Wolbachia surface protein might carry out this function.}, } @article {pmid16405591, year = {2006}, author = {Engelstädter, J and Hurst, GD}, title = {Can maternally transmitted endosymbionts facilitate the evolution of haplodiploidy?.}, journal = {Journal of evolutionary biology}, volume = {19}, number = {1}, pages = {194-202}, doi = {10.1111/j.1420-9101.2005.00974.x}, pmid = {16405591}, issn = {1010-061X}, mesh = {Animals ; *Biological Evolution ; Female ; Invertebrates/*genetics/*microbiology ; Male ; *Models, Biological ; *Ploidies ; Sex Ratio ; *Symbiosis ; Wolbachia/physiology ; }, abstract = {Whilst many invertebrate taxa are haplodiploid, the factors underlying the evolution of haplodiploidy remain unresolved. We investigate theoretically whether haplodiploidy might evolve as an outcome of the co-evolution between maternally inherited endosymbionts and their hosts. First, we substantially extend a recently developed model that involves maternally inherited endosymbionts that kill male offspring by eliminating the paternal genome. We also put forward a new hypothesis and develop a model that involves bacteria that induce cytoplasmic incompatibility (CI). Based on these models, we explore the co-evolutionary events that might occur between hosts and symbionts. We find that both with male-killers and CI-inducing endosymbionts, the hosts are likely to develop increased viability of haploid males, which can be considered a preadaptation to haplodiploidy. In addition, populations with haploidizing male-killers can in some cases evolve directly towards a genetic system of paternal genome elimination, a special form of haplodiploidy. These results are combined with consideration of mechanism and ecology to appraise the likelihood of male-killers and CI inducing bacteria being involved in the evolution of haplodiploidy.}, } @article {pmid16396448, year = {2005}, author = {Zakharov, IA and Markov, AV}, title = {[Gene orders in genomes of alpha-proteobacteria: similarity and evolution].}, journal = {Genetika}, volume = {41}, number = {12}, pages = {1624-1633}, pmid = {16396448}, issn = {0016-6758}, mesh = {Bacteria/*genetics ; *Evolution, Molecular ; Genes, Bacterial/*genetics ; *Phylogeny ; Sequence Homology, Nucleic Acid ; }, abstract = {The gene orders in the genomes of nine alpha-proteobacteria were compared using quantitative indices S (the relative number of common pairs of adjacent genes) and L (the mean difference between intergenic distances). A sample of 200 homologous genes, occurring in all 11 strains, was studied. In all of the genomes examined, 20 conserved, "uninterrupted" regions, including in total 63 out of 200 genes, were found. The rate of evolutionary change in the gene order widely varied in different evolutionary lineages. The highest rate (40 to 60 genome rearrangements per 100 Myr) was characteristic of the intercellular parasite Wolbachia (Rickettsiales). Computer simulation has showed that the S to L ratio observed in the sample testified that the probability of large genome rearrangements was somewhat lower than that of small ones.}, } @article {pmid16396396, year = {2005}, author = {Vinogradova, EB and Shaĭkevich, EV}, title = {[Differentiation between the urban mosquito Culex pipiens pipiens F. molestus and Culex torrentium (Diptera: Culicidae) by the molecular genetic methods].}, journal = {Parazitologiia}, volume = {39}, number = {6}, pages = {574-576}, pmid = {16396396}, issn = {0031-1847}, mesh = {Animals ; Culex/*classification/genetics/microbiology ; DNA, Mitochondrial/genetics ; DNA, Ribosomal Spacer/genetics ; Electron Transport Complex IV/genetics ; Genome, Insect/genetics ; Restriction Mapping ; Species Specificity ; Wolbachia/isolation & purification ; }, abstract = {The molecular characters suitable for the differentiation between the autogenous form of Culex pipiens pipiens (the urban mosquito) and the species Culex torrentium, which was previously identified erroneously as an anautogenous form of C. pipiens pipiens (Shaikevich, Vinogradova, 2004), are given. These mosquitoes differ in the infection by the endosymbiotic bacteria Wolbachia pipientis, 6 transitions among 247 nucleotide sequences of 3' part of the cytochrome oxidase subunit I (COI) gene of mitochondrial DNA (mtDNA), in the mtDNA fragments resulted from the restriction analysis, and in the lengths of second internal transcribed spacer (ITS2) sequences in the ribosomal DNA.}, } @article {pmid16393353, year = {2006}, author = {Mouton, L and Henri, H and Bouletreau, M and Vavre, F}, title = {Effect of temperature on Wolbachia density and impact on cytoplasmic incompatibility.}, journal = {Parasitology}, volume = {132}, number = {Pt 1}, pages = {49-56}, doi = {10.1017/S0031182005008723}, pmid = {16393353}, issn = {0031-1820}, mesh = {Analysis of Variance ; Animals ; Diploidy ; Female ; Genotype ; Haploidy ; Host-Parasite Interactions ; Male ; Polymerase Chain Reaction/methods ; Population Density ; Reproduction/*physiology ; Sex Ratio ; Species Specificity ; Symbiosis/*physiology ; *Temperature ; Wasps/*microbiology ; Wolbachia/*growth & development ; }, abstract = {The outcome and the evolution of host-symbiont associations depend on environmental constraints, but responses are difficult to predict since they arise from a complex interaction between the host, the parasite and the environment. The situation can be even more complex when multiple parasite genotypes, with potentially different responses to environmental changes, coexist within a single host. In this paper, we investigated the effect of the temperature (from 14 to 26 degrees C) during the host development on the density of 3 strains of the intracellular bacterium Wolbachia that coexist within the wasp Leptopilina heterotoma. In this species, Wolbachia induces cytoplasmic incompatibility, a sperm-egg incompatibility that allows it to spread and persist in host populations. Using real-time quantitative PCR we found that (i) Wolbachia density is temperature-specific and highest at 26 degrees C; (ii) the order of the abundance of the 3 Wolbachia strains does not vary with temperature changes; (iii) the response of bacterial density to temperature occurs within a single insect generation, during the egg-to-adult developmental period; (iv) in this species, temperature-related changes in Wolbachia density do not influence cytoplasmic incompatibility.}, } @article {pmid16391124, year = {2006}, author = {Miller, WJ and Riegler, M}, title = {Evolutionary dynamics of wAu-like Wolbachia variants in neotropical Drosophila spp.}, journal = {Applied and environmental microbiology}, volume = {72}, number = {1}, pages = {826-835}, pmid = {16391124}, issn = {0099-2240}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Drosophila/classification/genetics/*microbiology ; *Evolution, Molecular ; *Genetic Variation ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; *Tropical Climate ; Wolbachia/classification/*genetics/*isolation & purification/pathogenicity ; }, abstract = {Wolbachia bacteria are common intracellular symbionts of arthropods and have been extensively studied in Drosophila. Most research focuses on two Old Word hosts, Drosophila melanogaster and Drosophila simulans, and does not take into account that some of the Wolbachia associations in these species may have evolved only after their fast global expansion and after the exposure to Wolbachia of previously isolated habitats. Here we looked at Wolbachia of Neotropical Drosophila species. Seventy-one lines of 16 Neotropical Drosophila species sampled in different regions and at different time points were analyzed. Wolbachia is absent in lines of Drosophila willistoni collected before the 1970s, but more recent samples are infected with a strain designated wWil. Wolbachia is absent in all other species of the willistoni group. Polymorphic wWil-related strains were detected in some saltans group species, with D. septentriosaltans being coinfected with at least four variants. Based on wsp and ftsZ sequence data, wWil of D. willistoni is identical to wAu, a strain isolated from D. simulans, but can be discriminated when using a polymorphic minisatellite marker. In contrast to wAu, which infects both germ line and somatic tissues of D. simulans, wWil is found exclusively in the primordial germ line cells of D. willistoni embryos. We report on a pool of closely related Wolbachia strains in Neotropical Drosophila species as a potential source for the wAu strain in D. simulans. Possible evolutionary scenarios reconstructing the infection history of wAu-like Wolbachia in Neotropical Drosophila species and the Old World species D. simulans are discussed.}, } @article {pmid19537991, year = {2006}, author = {Dong, P and Wang, JJ and Zhao, ZM}, title = {Infection by Wolbachia bacteria and its influence on the reproduction of the stored-product psocid, Liposcelis tricolor.}, journal = {Journal of insect science (Online)}, volume = {6}, number = {}, pages = {1-7}, pmid = {19537991}, issn = {1536-2442}, mesh = {Animals ; Female ; Insecta/*microbiology/*physiology ; Male ; Molecular Sequence Data ; Ovum ; Phylogeny ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacteria that infect a wide range of arthropods and nematodes and are associated with various reproductive abnormalities in their hosts. The infection by Wolbachia of the psocid, Liposcelis tricolor (Psocoptera: Liposcelididae), was investigated using long PCR amplification of the wsp gene that codes for a Wolbachia surface protein. The results showed that L. tricolor was positive for Wolbachia . Phylogenetic analysis showed that the Wolbachia found in L. tricolor was related to the B-group. Wolbachia infection in L. tricolor could be removed through antibiotic treatment. The results of crosses including female(W+) x male(W+), female(W-) x male(W+), female(W+) x male(W-), and female(W-) x male(W-), suggested that the removal of Wolbachia resulted in lower egg production by L. tricolor . The mean embryonic mortality of offspring produced by L. tricolor without Wolbachia was significantly higher than that of control.}, } @article {pmid16365997, year = {2005}, author = {Sanogo, YO and Eitam, A and Dobson, SL}, title = {No evidence for bacteriophage WO orf7 correlation with Wolbachia-induced cytoplasmic incompatibility in the Culex pipiens complex (Culicidae: Diptera).}, journal = {Journal of medical entomology}, volume = {42}, number = {5}, pages = {789-794}, doi = {10.1093/jmedent/42.5.789}, pmid = {16365997}, issn = {0022-2585}, support = {AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/*genetics ; Crosses, Genetic ; Culex/genetics/*microbiology ; *Genetics, Population ; Open Reading Frames/genetics ; *Phenotype ; *Phylogeny ; Reproduction/genetics ; Sequence Analysis, DNA ; Wolbachia/*virology ; }, abstract = {Gene flow between populations of Culex pipiens L. is relevant to observed differences in disease transmission, insecticide resistance, behavior, and physiology. Intracellular Wolbachia bacteria have been hypothesized to affect gene flow in insects. Specifically, Wolbachia cause a form of embryonic mortality known as cytoplasmic incompatibility (CI) in crosses between individuals with different Wolbachia types. Incompatibility in Culex is exceptional in that it represents the most complex CI pattern known, and yet Culex populations are not infected with divergent Wolbachia strains. This has led to the hypothesis that extrachromosomal factors such as phages or mobile genetic elements may be involved in determining CI phenotype. Recent molecular characterization of Culex laboratory strains has identified variation in the orf7 locus of the Wolbachia-associated bacteriophage WO. Here, crosses between eight Culex strains differing in their orf7 type were conducted to examine for the hypothesized involvement of bacteriophage WO in determining CI in Culex. Although crossing results show examples of compatibility, partial compatibility, and incompatibility, the results fail to show a correlation between the CI phenotypes and orf7 type. Specific examples include high egg hatch resulting in crosses between Culex strains that differ significantly in their orf7 type and low egg hatch resulting in crosses between Culex strains with similar orf7 types. Thus, the phage orf7 locus alone cannot predict CI type in the Culex strains examined in this study. However, rejection of the hypothesized role of WO phage in Culex CI will require the characterization of additional phage loci.}, } @article {pmid16361236, year = {2006}, author = {Engelstädter, J and Charlat, S and Pomiankowski, A and Hurst, GD}, title = {The evolution of cytoplasmic incompatibility types: integrating segregation, inbreeding and outbreeding.}, journal = {Genetics}, volume = {172}, number = {4}, pages = {2601-2611}, pmid = {16361236}, issn = {0016-6731}, mesh = {Bacteria/genetics ; Crosses, Genetic ; Cytoplasm/*metabolism ; Evolution, Molecular ; Genes, Bacterial ; Models, Genetic ; Models, Statistical ; Mutation ; Wolbachia/genetics ; }, abstract = {Cytoplasmic incompatibility (CI) is a reproductive incompatibility induced by maternally transmitted bacteria of the genera Wolbachia and Cardinium. In the simplest form of CI, offspring from infected males and uninfected females suffer from increased mortality. However, it has been noted that crosses between males and females carrying different strains of infection are often also incompatible. The evolutionary processes leading to the emergence of new CI-compatibility types are still not resolved. Here, we develop a model that extends previous theoretical approaches by including segregation of bacterial strains during transmission as well as a continuum of breeding systems ranging from inbreeding (complete sib mating) to outbreeding (complete sib-mating avoidance). Our results demonstrate that (1) with segregation of strains, evolution is unlikely to lead to new CI types that co-occur as a double infection with the preexisting one, (2) inbreeding substantially hampers the evolution of new CI types, and (3) outbreeding facilitates the evolution of new CI types. Our model also provides a hypothesis on the evolutionary origin of CI.}, } @article {pmid16360953, year = {2006}, author = {Sanogo, YO and Dobson, SL}, title = {WO bacteriophage transcription in Wolbachia-infected Culex pipiens.}, journal = {Insect biochemistry and molecular biology}, volume = {36}, number = {1}, pages = {80-85}, doi = {10.1016/j.ibmb.2005.11.001}, pmid = {16360953}, issn = {0965-1748}, support = {AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteriophages/*genetics ; Culex/classification/*microbiology ; DNA, Viral ; Female ; *Gene Expression Regulation, Viral ; Genetic Variation ; Life Cycle Stages ; Male ; RNA, Viral ; Sex Factors ; Transcription, Genetic ; Wolbachia/*virology ; }, abstract = {Bacteriophages are commonly found in association with free-living bacteria, both as exogenic phages (virions) and as prophages integrated into the bacterial genome. In contrast, the observation of bacteriophages associated with obligate intracellular bacteria has been described infrequently. An exception is provided by Wolbachia endosymbionts, which harbor multiple phage elements that have been designated as WO phage. Wolbachia are maternally inherited bacteria that occur in the cytoplasm of many invertebrates, where they often manipulate host reproduction. Previously, the WO phage orf7 locus and ankyrin repeat-encoding genes have been observed to represent sources of genetic diversity between Wolbachia (wPip) strains infecting mosquitoes of the Culex pipiens complex and have been suggested as potential participants in the reproductive manipulations. We have characterized WO phage associated with multiple Wolbachia-infected Culex strains and an uninfected strain using electron microscopy and RT-PCR. For each strain, different developmental stages were examined for transcription of three WO phage orf7 genes. The results provide evidence for the presence of both actively transcribed virions and inactive prophages. Variable orf7 transcription patterns are observed in comparisons of differing Cx. pipiens strains. Variability includes both mosquito stage-specific and sexually dimorphic orf7 expression patterns. This report provides additional support for the hypothesis that bacteriophages play an important role in Wolbachia and host evolution.}, } @article {pmid16342786, year = {2005}, author = {Chu, D and Zhang, YJ and Bi, YP and Fu, HB}, title = {[Wolbachia endosymbionts and their effects on the fitness of the arthropod hosts].}, journal = {Wei sheng wu xue bao = Acta microbiologica Sinica}, volume = {45}, number = {5}, pages = {817-820}, pmid = {16342786}, issn = {0001-6209}, mesh = {Animals ; Arthropods/*microbiology/physiology ; Ecology ; Reproduction ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia are common and maternally inherited bacteria found in reproductive tissue of a wide range of arthropod species. A tremendous amount of progress on their manipulating reproduction of their host has been made over the past 30 years. Recent surveys have found that they could effect the fitness of their hosts. The recent advances on Wolbachia distribution, locality and their effects on the fitness of hosts are reviewed, and the significance and potential implications of the fields are discussed.}, } @article {pmid16339946, year = {2005}, author = {Casiraghi, M and Bordenstein, SR and Baldo, L and Lo, N and Beninati, T and Wernegreen, JJ and Werren, JH and Bandi, C}, title = {Phylogeny of Wolbachia pipientis based on gltA, groEL and ftsZ gene sequences: clustering of arthropod and nematode symbionts in the F supergroup, and evidence for further diversity in the Wolbachia tree.}, journal = {Microbiology (Reading, England)}, volume = {151}, number = {Pt 12}, pages = {4015-4022}, doi = {10.1099/mic.0.28313-0}, pmid = {16339946}, issn = {1350-0872}, support = {R01 GM62626-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics ; Chaperonin 60/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/genetics ; Filarioidea/*microbiology ; Genes, Bacterial ; Insecta/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/chemistry ; Symbiosis ; Wolbachia/*classification/genetics ; }, abstract = {Current phylogenies of the intracellular bacteria belonging to the genus Wolbachia identify six major clades (A-F), termed 'supergroups', but the branching order of these supergroups remains unresolved. Supergroups A, B and E include most of the wolbachiae found thus far in arthropods, while supergroups C and D include most of those found in filarial nematodes. Members of supergroup F have been found in arthropods (i.e. termites), and have previously been detected in the nematode Mansonella ozzardi, a causative agent of human filariasis. To resolve the phylogenetic positions of Wolbachia from Mansonella spp., and other novel strains from the flea Ctenocephalides felis and the filarial nematode Dipetalonema gracile, the authors generated new DNA sequences of the Wolbachia genes encoding citrate synthase (gltA), heat-shock protein 60 (groEL), and the cell division protein ftsZ. Phylogenetic analysis confirmed the designation of Wolbachia from Mansonella spp. as a member of the F supergroup. In addition, it was found that divergent lineages from Dip. gracile and Cte. felis lack any clear affiliation with known supergroups, indicating further genetic diversity within the Wolbachia genus. Finally, although the data generated did not permit clear resolution of the root of the global Wolbachia tree, the results suggest that the transfer of Wolbachia spp. from arthropods to nematodes (or vice versa) probably occurred more than once.}, } @article {pmid16332874, year = {2005}, author = {Dunn, AK and Stabb, EV}, title = {Culture-independent characterization of the microbiota of the ant lion Myrmeleon mobilis (Neuroptera: Myrmeleontidae).}, journal = {Applied and environmental microbiology}, volume = {71}, number = {12}, pages = {8784-8794}, pmid = {16332874}, issn = {0099-2240}, mesh = {Alphaproteobacteria/classification/*isolation & purification ; Animal Feed ; Animals ; Betaproteobacteria/classification/*isolation & purification ; Digestive System/microbiology ; Enterobacteriaceae/classification/isolation & purification ; Gammaproteobacteria/classification/*isolation & purification ; Insecta/*microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/*genetics ; }, abstract = {Ant lions are insect larvae that feed on the liquefied internal components of insect prey. Prey capture is assisted by the injection of toxins that are reportedly derived from both the insect and bacterial symbionts. These larvae display interesting gut physiology where the midgut is not connected to the hindgut, preventing elimination of solid waste until adulthood. The presence of a discontinuous gut and the potential involvement of bacteria in prey paralyzation suggest an interesting microbial role in ant lion biology; however, the ant lion microbiota has not been described in detail. We therefore performed culture-independent 16S rRNA gene sequence analysis of the bacteria associated with tissues of an ant lion, Myrmeleon mobilis. All 222 sequences were identified as Proteobacteria and could be subdivided into two main groups, the alpha-Proteobacteria with similarity to Wolbachia spp. (75 clones) and the gamma-Proteobacteria with similarity to the family Enterobacteriaceae (144 clones). The Enterobacteriaceae-like 16S rRNA gene sequences were most commonly isolated from gut tissue, and Wolbachia-like sequences were predominant in the head and body tissue. Fluorescence in situ hybridization analyses supported the localization of enterics to gut tissue and Wolbachia to nongut tissue. The diversity of sequences isolated from freshly caught, laboratory-fed, and laboratory-starved ant lions were qualitatively similar, although the libraries from each treatment were significantly different (P = 0.05). These results represent the first culture-independent analysis of the microbiota associated with a discontinuous insect gut and suggest that the ant lion microbial community is relatively simple, which may be a reflection of the diet and gut physiology of these insects.}, } @article {pmid16329235, year = {2005}, author = {Telschow, A and Hammerstein, P and Werren, JH}, title = {The effect of Wolbachia versus genetic incompatibilities on reinforcement and speciation.}, journal = {Evolution; international journal of organic evolution}, volume = {59}, number = {8}, pages = {1607-1619}, pmid = {16329235}, issn = {0014-3820}, mesh = {Animals ; Computer Simulation ; *Genetics, Population ; Insecta/genetics/*microbiology/*physiology ; *Models, Biological ; Population Dynamics ; Reproduction/genetics/physiology ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a widespread group of intracellular bacteria commonly found in arthropods. In many insect species, Wolbachia induce a cytoplasmic mating incompatibility (CI). If different Wolbachia infections occur in the same host species, bidirectional CI is often induced. Bidirectional CI acts as a postzygotic isolation mechanism if parapatric host populations are infected with different Wolbachia strains. Therefore, it has been suggested that Wolbachia could promote speciation in their hosts. In this article we investigate theoretically whether Wolbachia-induced bidirectional CI selects for premating isolation and therefore reinforces genetic divergence between parapatric host populations. To achieve this we combined models for Wolbachia dynamics with a well-studied reinforcement model. This new model allows us to compare the effect of bidirectional CI on the evolution of female mating preferences with a situation in which postzygotic isolation is caused by nuclear genetic incompatibilities (NI). We distinguish between nuclear incompatibilities caused by two loci with epistatic interactions, and a single locus with incompatibility among heterozygotes in the diploid phase. Our main findings are: (1) bidirectional CI and single locus NI select for premating isolation with a higher speed and for a wider parameter range than epistatic NI; (2) under certain parameter values, runaway sexual selection leads to the increase of an introduced female preference allele and fixation of its preferred male trait allele in both populations, whereas under others it leads to divergence in the two populations in preference and trait alleles; and (3) bidirectional CI and single locus NI can stably persist up to migration rates that are two times higher than seen for epistatic NI. The latter finding is important because the speed with which mutants at the preference locus spread increases exponentially with the migration rate. In summary, our results show that bidirectional CI selects for rapid premating isolation and so generally support the view that Wolbachia can promote speciation in their hosts.}, } @article {pmid16313565, year = {2005}, author = {van Opijnen, T and Baudry, E and Baldo, L and Bartos, J and Werren, JH}, title = {Genetic variability in the three genomes of Nasonia: nuclear, mitochondrial and Wolbachia.}, journal = {Insect molecular biology}, volume = {14}, number = {6}, pages = {653-663}, doi = {10.1111/j.1365-2583.2005.00595.x}, pmid = {16313565}, issn = {0962-1075}, mesh = {Animals ; Cell Nucleus/*genetics ; DNA, Mitochondrial/*genetics ; *Genetic Variation ; *Genome, Bacterial ; *Genome, Insect ; Phylogeny ; Wasps/cytology/*genetics/microbiology ; Wolbachia/*genetics ; }, abstract = {Nasonia consists of three closely related species of parasitoid wasps that are all infected with the endosymbiotic bacteria Wolbachia, a reproductive parasite common in arthropods. This situation presents the opportunity to compare patterns of variation in three associated genomes, Wolbachia and the nuclear and mitochondrial genomes of its host. Furthermore, although Nasonia wasps are emerging as a model for evolutionary and genetic studies, little is known about their genetic variability. Using amplified fragment length polymorphisms (AFLPs), all three species present a relatively high level of nuclear polymorphism and have different patterns of variation, with one of the species, Nasonia giraulti, being divided into two divergent subgroups. In each species, the mitochondrial pattern of variation is different from the nuclear pattern, possibly due to genetic hitchhiking of the mitochondria during (cytoplasmically inherited) Wolbachia sweeps. Mitochondria in Nasonia show a synonymous substitution rate approximately 10-15-fold higher than nuclear genes, probably reflecting an elevated mitochondrial mutation rate that is among the highest found in insects. Finally, all three species are doubly infected with their own strains of Wolbachia, one each from the two major supergroups (A and B). Sequence analysis reveals that each of the three Nasonia species acquired their A and B bacteria independently by horizontal transfer events from other insects with the exception of B type Wolbachia in N. longicornis and N. giraulti, which were acquired prior to speciation and then codiverged with the host. This represents one of the few clear-cut examples of codivergence of Wolbachia during host speciation.}, } @article {pmid16304620, year = {2005}, author = {Puttaraju, HP and Prakash, BM}, title = {Wolbachia and reproductive conflict in Exorista sorbillans.}, journal = {Archives of insect biochemistry and physiology}, volume = {60}, number = {4}, pages = {230-235}, doi = {10.1002/arch.20069}, pmid = {16304620}, issn = {0739-4462}, mesh = {Analysis of Variance ; Animals ; Anti-Bacterial Agents/toxicity ; Bombyx/*parasitology ; Crosses, Genetic ; Diptera/*microbiology/*physiology ; Fertility/physiology ; Host-Parasite Interactions/physiology ; Reproduction/physiology ; Sex Ratio ; *Symbiosis ; Wolbachia/drug effects/*physiology ; }, abstract = {Many arthropods harbour endosymbiotic bacteria of the genus Wolbachia. These endosymbionts are transmitted vertically from one generation to the next and are obligatory in several Dipterans that have been studied to date. These bacteria induce an array of reproductive isolation mechanisms that are implicated in pest management to evolutionary biology of respective hosts. The uzifly, Exorista sorbillans, a tachinid endoparasitoid of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), causes enormous losses to the silk industry; now it is known that it harbours Wolbachia endobacteria. The elimination of Wolbachia by antibiotics interrupts embryogenesis and causes various reproductive conflicts such as (1) a reduction of fecundity of uninfected female, (2) cytoplasmic incompatibility in the uninfected females crossed with infected males, (3) genomic incompatibility in crosses between males and females from uninfected population, and (4) sex-ratio distortion in uninfected females irrespective of the presence of Wolbachia in males. These results suggest that the relationship of Wolbachia with its uzifly host is one of mutual symbiosis as it controls the reproductive physiology of its host.}, } @article {pmid16299326, year = {2005}, author = {Kerepesi, LA and Leon, O and Lustigman, S and Abraham, D}, title = {Protective immunity to the larval stages of onchocerca volvulus is dependent on Toll-like receptor 4.}, journal = {Infection and immunity}, volume = {73}, number = {12}, pages = {8291-8297}, pmid = {16299326}, issn = {0019-9567}, support = {R01 AI047189/AI/NIAID NIH HHS/United States ; AI042328/AI/NIAID NIH HHS/United States ; AI47189/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Antibody Formation ; Helminth Proteins/immunology ; Interferon-gamma/metabolism ; Larva/immunology/ultrastructure ; Mice ; Mice, Mutant Strains ; Onchocerca volvulus/growth & development/*immunology/microbiology ; Onchocerciasis/genetics/*immunology ; Spleen/cytology ; Symbiosis ; Th2 Cells/immunology ; Toll-Like Receptor 4/genetics/*physiology ; Wolbachia/immunology/ultrastructure ; }, abstract = {Toll-like receptor 4 (TLR4) has been shown to be important for the induction of Th2-dependent immune responses in mice. Protective immunity against larval Onchocerca volvulus in mice depends on the development of a Th2 immune response mediated by both interleukin-4 (IL-4) and IL-5. In addition, O. volvulus contains the rickettsial endosymbiont Wolbachia, which has molecules with lipopolysaccharide-like activities that also signal through TLR4. We therefore hypothesized that protective immunity to O. volvulus would not develop in C3H/HeJ mice which have a mutation in the Tlr4 gene (TLR4 mutant), either because of a decreased Th2 response to the larvae or because of the absence of a response to Wolbachia. TLR4-mutant mice were immunized against O. volvulus with irradiated third-stage larvae, and it was observed that Th2 responses were elevated based on increased IL-5 production, total immunoglobulin E (IgE) levels, antigen-specific IgG1 response, and eosinophil recruitment. Protective immunity, however, did not develop in the TLR4-mutant mice. The Th1 response, as measured by gamma interferon production from spleen cells, was comparable in both wild-type and TLR4-mutant mice. Furthermore, antibody responses to Wolbachia were absent in both wild-type and TLR4-mutant mice. Therefore, the defect in the development of a protective immune response against O. volvulus in TLR4-mutant mice is not due to loss of Th2 immunity or the response to Wolbachia but is due to an unidentified TLR4-dependent larval killing mechanism.}, } @article {pmid16278157, year = {2005}, author = {Bajpai, P and Vedi, S and Owais, M and Sharma, SK and Saxena, PN and Misra-Bhattacharya, S}, title = {Use of liposomized tetracycline in elimination of Wolbachia endobacterium of human lymphatic filariid Brugia malayi in a rodent model.}, journal = {Journal of drug targeting}, volume = {13}, number = {6}, pages = {375-381}, doi = {10.1080/10611860500285439}, pmid = {16278157}, issn = {1061-186X}, mesh = {Administration, Oral ; Animals ; Anti-Bacterial Agents/*administration & dosage/therapeutic use ; Brugia malayi/*drug effects ; Disease Models, Animal ; Elephantiasis, Filarial/*drug therapy ; Female ; Humans ; Liposomes ; Rats ; Tetracycline/*administration & dosage/therapeutic use ; Wolbachia/*drug effects/pathogenicity ; }, abstract = {Wolbachia bacteria, being filarial parasite symbiont have been implicated in a variety of roles, including development, fecundity and the pathogenesis of the filarial infections. Among various strategies used in the treatment of experimental filariasis, the elimination of symbiont Wolbachia seem to offer an efficient means of curing the disease. The antiwolbachial property of tetracycline has been well worked out; however, treatment needs to be continued for a prolonged period of time to achieve complete elimination of Wolbachia from the filarial parasites and their subsequent killing. This results in acute toxicity, thus limiting its practical utility for clinical implementation. In order to increase efficacy of the antibiotic with minimal toxic manifestations, we developed liposomized formulation of the tetracycline. The liposomized tetracycline was found to be significantly more effective when compared to the free form of the drug. In contrast to the 90/120 days oral administration of the drug, the treatment schedule using the liposomized form of the drug was reduced to 12 alternate days with better efficacy of the treatment.}, } @article {pmid16274474, year = {2005}, author = {Sahu, BR and Mohapatra, AD and Majumder, A and Das, PK and Ravindran, B}, title = {A flow cytometry based method for studying embryogenesis and immune reactivity to embryogenic stages in filarial parasites.}, journal = {Filaria journal}, volume = {4}, number = {1}, pages = {11}, pmid = {16274474}, issn = {1475-2883}, abstract = {BACKGROUND: In the absence of intermediate animal hosts, the process of embryogenesis leading to fecundity of adult female filarial worms is very critical for persistence of these obligate parasites in human communities. Embryogenesis in adult female filarial parasites involves fertilization of eggs or oocytes by sperms and their subsequent development into motile microfilariae inside the uterine cavity of worms. Development of assays for monitoring embryogenesis in adult female worms is a critical requirement in filariasis research--filarial worms are known to harbour endosymbionts such as Wolbachia which play a significant role in fecundity. Tetracycline or doxycycline treatment of the infected hosts effectively eliminates the endosymbionts resulting in inhibition of embryogenesis in female worms. Currently, inhibition of embryogenesis in adult filarial worms can be monitored only by microscopic examination of in vitro harvested intrauterine stages.

METHODS: Adult female filarial worms of bovine filarial parasites, Setaria digitata were collected from the peritoneum of infected animals and intrauterine stages were harvested in culture medium and were analyzed for forward and side scatter by flowcytometry using a BD FACS Calibur. Different populations were gated, sorted and identified by phase microscopy. Binding of biotinylated lectins to intra uterine stages was monitored using FITC labeled Avidin and monitored by flow cytometry of gated populations. Similarly, binding of antibodies in human filarial sera to intrauterine stages was monitored using FITC labeled anti-human immunoglobulins.

RESULTS: The forward and side scatter for intrauterine stages delineated 3 distinct populations labeled as R1, R2 and R3. The three populations were sorted and identified to be a) fully stretched microfilariae, b) early and c) late developmental stages of eggs respectively. Lectins such as Wheat Germ agglutinin or Concanavalin-A were found to bind strongly to egg stages and less prominently to intra-uterine microfilariae. Similarly the binding of antibodies in filarial sera to the three intra-uterine stages could also be precisely quantified.

CONCLUSION: The manuscript reports a novel flow cytometry based method to monitor progression of embryogenesis in adult filarial worms. Apart from relative quantification of different intra uterine developmental stages, the assay allows quantitative binding of lectins and antibodies to each of the intrauterine stages. It may now be possible to quantify levels of antibodies in infected and immune hosts to monitor anti-fecundity immunity in filariasis--the assay can thus be used as a powerful tool for drug development and in immunological studies in human and experimental filariasis.}, } @article {pmid16269725, year = {2005}, author = {Mavingui, P and Van, VT and Labeyrie, E and Rancès, E and Vavre, F and Simonet, P}, title = {Efficient procedure for purification of obligate intracellular Wolbachia pipientis and representative amplification of its genome by multiple-displacement amplification.}, journal = {Applied and environmental microbiology}, volume = {71}, number = {11}, pages = {6910-6917}, pmid = {16269725}, issn = {0099-2240}, mesh = {Animals ; Bacteriological Techniques ; Cell Line ; DNA, Bacterial/*analysis/*isolation & purification ; Drosophila/growth & development/*microbiology ; Electrophoresis, Gel, Pulsed-Field ; *Genome, Bacterial ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques/*methods ; Ovum/microbiology ; Sequence Analysis, DNA ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Bacteria belonging to the genus Wolbachia are obligatory microendocytobionts that infect a variety of arthropods and a majority of filarial nematode species, where they induce reproductive alterations or establish a mutualistic symbiosis. Although two whole genome sequences of Wolbachia pipientis, for strain wMel from Drosophila melanogaster and strain wBm from Brugia malayi, have been fully completed and six other genome sequencing projects are ongoing (http://www.genomesonline.org/index.cgi?want=Prokaryotic+Ongoin), genetic analyses of these bacteria are still scarce, mainly due to the inability to cultivate them outside of eukaryotic cells. Usually, a large amount of host tissue (a thousand individuals, or about 10 g) is required in order to purify Wolbachia and extract its DNA, which is often recovered in small amounts and contaminated by host cell DNA, thus hindering genomic studies. In this report, we describe an efficient and reliable procedure to representatively amplify the Wolbachia genome by multiple-displacement amplification from limited infected host tissue (0.2 g or 2 x 10(7) cells). We obtained sufficient amounts (8 to 10 microg) of DNA of suitable quality for genomic studies, and we demonstrated that the amplified DNA contained all of the Wolbachia loci targeted. In addition, our data indicated that the genome of strain wRi, an obligatory endosymbiont of Drosophila simulans, shares a similar overall architecture with its relative strain wMel.}, } @article {pmid16267140, year = {2006}, author = {Baldo, L and Bordenstein, S and Wernegreen, JJ and Werren, JH}, title = {Widespread recombination throughout Wolbachia genomes.}, journal = {Molecular biology and evolution}, volume = {23}, number = {2}, pages = {437-449}, doi = {10.1093/molbev/msj049}, pmid = {16267140}, issn = {0737-4038}, support = {R01 GM62626-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Bacterial Proteins/*genetics ; Bacteriophages/genetics ; DNA Transposable Elements/genetics ; Gene Transfer, Horizontal/*genetics ; Genome, Bacterial/*genetics ; Wolbachia/*genetics ; }, abstract = {Evidence is growing that homologous recombination is a powerful source of genetic variability among closely related free-living bacteria. Here we investigate the extent of recombination among housekeeping genes of the endosymbiotic bacteria Wolbachia. Four housekeeping genes, gltA, dnaA, ftsZ, and groEL, were sequenced from a sample of 22 strains belonging to supergroups A and B. Sequence alignments were searched for recombination within and between genes using phylogenetic inference, analysis of genetic variation, and four recombination detection programs (MaxChi, Chimera, RDP, and Geneconv). Independent analyses indicate no or weak intragenic recombination in ftsZ, dnaA, and groEL. Intragenic recombination affects gltA, with a clear evidence of horizontal DNA transfers within and between divergent Wolbachia supergroups. Intergenic recombination was detected between all pairs of genes, suggesting either a horizontal exchange of a genome portion encompassing several genes or multiple recombination events involving smaller tracts along the genome. Overall, the observed pattern is compatible with pervasive recombination. Such results, combined with previous evidence of recombination in a surface protein, phage, and IS elements, support an unexpected chimeric origin of Wolbachia strains, with important implications for Wolbachia phylogeny and adaptation of these obligate intracellular bacteria in arthropods.}, } @article {pmid16252129, year = {2005}, author = {Bordenstein, S and Rosengaus, RB}, title = {Discovery of a novel Wolbachia super group in Isoptera.}, journal = {Current microbiology}, volume = {51}, number = {6}, pages = {393-398}, pmid = {16252129}, issn = {0343-8651}, mesh = {Animals ; Bacterial Proteins/genetics ; Chaperonin 60/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Genetic Variation ; Isoptera/*microbiology ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Wolbachia are one of the most abundant groups of bacterial endosymbionts in the biosphere. Interest in these heritable microbes has expanded with the discovery of wider genetic diversity in under-sampled host species. Here, we report on the putative discovery of a new genetic lineage, denoted super group H, which infects the Isopteran species Zootermopsis angusticollis and Z. nevadensis. Evidence for this novel super group is based on portions of new Wolbachia gene sequences from each species spanning 3.5 kilobases of DNA and the following genes: 16S rDNA, dnaA, gltA, groEL, and ftsZ. Single-gene and concatenated maximum likelihood phylogenies establish this new super group and validate the positioning of the other Wolbachia super groups. This discovery is the first example of a termite Wolbachia that is highly divergent from the Isopteran Wolbachia previously described in super group F. This study highlights the importance of multilocus approaches to resolving Wolbachia super group relationships. It also suggests that surveys of Wolbachia in more earlier-originating (and under-sampled) groups of arthropods are more apt to reveal novel genetic diversity.}, } @article {pmid16251775, year = {2005}, author = {Rao, RU}, title = {Endosymbiotic Wolbachia of parasitic filarial nematodes as drug targets.}, journal = {The Indian journal of medical research}, volume = {122}, number = {3}, pages = {199-204}, pmid = {16251775}, issn = {0971-5916}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; Doxycycline/*pharmacology/therapeutic use ; Filariasis/*drug therapy/*epidemiology ; Humans ; India/epidemiology ; Reproduction/drug effects ; *Symbiosis ; Wolbachia/*drug effects ; Wuchereria bancrofti/*microbiology ; }, abstract = {The parasitic nematodes Wuchereria bancrofti, Brugia malayi and B. timori cause a dreadful disease in humans known as lymphatic filariasis, which afflicts more than 120 million people worldwide. As per recent epidemiologic estimates on prevalence of W. bancrofti and B. malayi, about 428 million people are at risk, with 28 million microfilaria carriers and 21 million clinical cases spread out in 13 States and 5 Union Territories of India. The Indian subcontinent that comprises Bangladesh, India, Maldives, Nepal and Sri Lanka harbours 50 per cent of the world's lymphatic filarial disease burden. Recently, an endobacterium of Wolbachia species that belongs to the family Rickettsiaceae was found in all life cycle stages of these nematodes and the transmission is exclusively vertical through the embryonic stages of the female worms. People with filariasis have been exposed to these Wolbachia bacteria or their proteins by the natural killing of parasites. Wolbachia have also been identified occasionally in body fluids of infected patients. Evidence suggests that these Wolbachia are mutualistic symbionts and can be cured from the nematodes by several antibiotics having antirickettsial properties. Treatment of nematodes with tetracyclines affect Wolbachia and they get cleared from worm tissues; and this elimination causes reproductive abnormalities in worms and affect worm's embryogenesis, resulting in sterility. Although it is impractical, prolonged treatment with doxycycline significantly reduces the numbers of microfilaria in circulation, which is an important strategy to control transmission of filariasis by mosquito vectors. In this review, the current knowledge of Wolbachia as a drug target and potential ways to reduce the infection through anti-Wolbachia treatments is discussed.}, } @article {pmid16230105, year = {2005}, author = {Taylor, MJ and Bandi, C and Hoerauf, A}, title = {Wolbachia bacterial endosymbionts of filarial nematodes.}, journal = {Advances in parasitology}, volume = {60}, number = {}, pages = {245-284}, doi = {10.1016/S0065-308X(05)60004-8}, pmid = {16230105}, issn = {0065-308X}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Antibodies, Bacterial/biosynthesis ; Arthropods/*microbiology ; Dogs ; Doxycycline/therapeutic use ; Environment ; *Filariasis/drug therapy/microbiology/parasitology ; Genomics ; Humans ; Inflammation Mediators/metabolism ; Nematoda/*microbiology ; Phylogeny ; Population Dynamics ; *Symbiosis ; Wolbachia/classification/drug effects/growth & development/*physiology ; }, abstract = {Filarial nematodes are important helminth parasites of the tropics and a leading cause of global disability. They include species responsible for onchocerciasis, lymphatic filariasis and dirofilariasis. A unique feature of these nematodes is their dependency upon a symbiotic intracellular bacterium, Wolbachia, which is essential for normal development and fertility. Advances in our understanding of the symbiosis of Wolbachia bacteria with filarial nematodes have made rapid progress in recent years. Here we summarise our current understanding of the evolution of the symbiotic association together with insights into the functional basis of the interaction derived from genomic analysis. Also we discuss the contribution of Wolbachia to inflammatory-mediated pathogenesis and adverse reactions to anti-filarial drugs and describe the outcome of recent field trials using antibiotics as a promising new tool for the treatment of filarial infection and disease.}, } @article {pmid16228015, year = {2005}, author = {Ferree, PM and Frydman, HM and Li, JM and Cao, J and Wieschaus, E and Sullivan, W}, title = {Wolbachia utilizes host microtubules and Dynein for anterior localization in the Drosophila oocyte.}, journal = {PLoS pathogens}, volume = {1}, number = {2}, pages = {e14}, pmid = {16228015}, issn = {1553-7374}, mesh = {Animals ; Cell Differentiation/physiology ; Drosophila melanogaster/*microbiology ; Female ; Microtubules/*microbiology ; Molecular Sequence Data ; Oocytes/cytology/*microbiology/*physiology ; Oogenesis ; Wolbachia/growth & development/*pathogenicity/physiology ; }, abstract = {To investigate the role of the host cytoskeleton in the maternal transmission of the endoparasitic bacteria Wolbachia, we have characterized their distribution in the female germ line of Drosophila melanogaster. In the germarium, Wolbachia are distributed to all germ cells of the cyst, establishing an early infection in the cell destined to become the oocyte. During mid-oogenesis, Wolbachia exhibit a distinct concentration between the anterior cortex and the nucleus in the oocyte, where many bacteria appear to contact the nuclear envelope. Following programmed rearrangement of the microtubule network, Wolbachia dissociate from this anterior position and become dispersed throughout the oocyte. This localization pattern is distinct from mitochondria and all known axis determinants. Manipulation of microtubules and cytoplasmic Dynein and Dynactin, but not Kinesin-1, disrupts anterior bacterial localization in the oocyte. In live egg chambers, Wolbachia exhibit movement in nurse cells but not in the oocyte, suggesting that the bacteria are anchored by host factors. In addition, we identify mid-oogenesis as a period in the life cycle of Wolbachia in which bacterial replication occurs. Total bacterial counts show that Wolbachia increase at a significantly higher rate in the oocyte than in the average nurse cell, and that normal Wolbachia levels in the oocyte depend on microtubules. These findings demonstrate that Wolbachia utilize the host microtubule network and associated proteins for their subcellular localization in the Drosophila oocyte. These interactions may also play a role in bacterial motility and replication, ultimately leading to the bacteria's efficient maternal transmission.}, } @article {pmid16224027, year = {2005}, author = {Xi, Z and Khoo, CC and Dobson, SL}, title = {Wolbachia establishment and invasion in an Aedes aegypti laboratory population.}, journal = {Science (New York, N.Y.)}, volume = {310}, number = {5746}, pages = {326-328}, doi = {10.1126/science.1117607}, pmid = {16224027}, issn = {1095-9203}, support = {AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Crosses, Genetic ; Cytoplasm ; Female ; Insect Vectors/microbiology ; Male ; Pest Control, Biological ; Reproduction ; Wolbachia/*physiology ; }, abstract = {A proposed strategy to aid in controlling the growing burden of vector-borne disease is population replacement, in which a natural vector population is replaced by a population with a reduced capacity for disease transmission. An important component of such a strategy is the drive system, which serves to spread a desired genotype into the targeted field population. Endosymbiotic Wolbachia bacteria are potential transgene drivers, but infections do not naturally occur in some important mosquito vectors, notably Aedes aegypti. In this work, stable infections of wAlbB Wolbachia were established in A. aegypti and caused high rates of cytoplasmic incompatibility (that is, elimination of egg hatch). Laboratory cage tests demonstrated the ability of wAlbB to spread into an A. aegypti population after seeding of an uninfected population with infected females, reaching infection fixation within seven generations.}, } @article {pmid16215223, year = {2005}, author = {Arcà, B and Lombardo, F and Valenzuela, JG and Francischetti, IM and Marinotti, O and Coluzzi, M and Ribeiro, JM}, title = {An updated catalogue of salivary gland transcripts in the adult female mosquito, Anopheles gambiae.}, journal = {The Journal of experimental biology}, volume = {208}, number = {Pt 20}, pages = {3971-3986}, doi = {10.1242/jeb.01849}, pmid = {16215223}, issn = {0022-0949}, support = {//Intramural NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Anopheles/*genetics ; Chromosome Mapping ; Chromosomes/genetics ; Cluster Analysis ; Computational Biology ; DNA Transposable Elements ; Female ; *Gene Expression Profiling ; *Gene Expression Regulation ; Insect Proteins/chemistry/*genetics ; Molecular Sequence Data ; RNA, Messenger/genetics/metabolism ; Salivary Glands/*metabolism ; Transcription, Genetic/*genetics ; }, abstract = {Salivary glands of blood-sucking arthropods contain a variety of compounds that prevent platelet and clotting functions and modify inflammatory and immunological reactions in the vertebrate host. In mosquitoes, only the adult female takes blood meals, while both sexes take sugar meals. With the recent description of the Anopheles gambiae genome, and with a set of approximately 3000 expressed sequence tags from a salivary gland cDNA library from adult female mosquitoes, we attempted a comprehensive description of the salivary transcriptome of this most important vector of malaria transmission. In addition to many transcripts associated with housekeeping functions, we found an active transposable element, a set of Wolbachia-like proteins, several transcription factors, including Forkhead, Hairy and doublesex, extracellular matrix components and 71 genes coding for putative secreted proteins. Fourteen of these 71 proteins had matching Edman degradation sequences obtained from SDS-PAGE experiments. Overall, 33 transcripts are reported for the first time as coding for salivary proteins. The tissue and sex specificity of these protein-coding transcripts were analyzed by RT-PCR and microarray experiments for insight into their possible function. Notably, two gene products appeared to be differentially spliced in the adult female salivary glands, whereas 13 contigs matched predicted intronic regions and may include additional alternatively spliced transcripts. Most An. gambiae salivary proteins represent novel protein families of unknown function, potentially coding for pharmacologically or microbiologically active substances. Supplemental data to this work can be found at http://www.ncbi.nlm.nih.gov/projects/omes/index.html#Ag2.}, } @article {pmid16208939, year = {2005}, author = {Markov, AV and Zakharov, IA}, title = {[Sexual reproduction of insects is regulated by cytoplasmic bacteria].}, journal = {Ontogenez}, volume = {36}, number = {4}, pages = {280-291}, pmid = {16208939}, issn = {0475-1450}, mesh = {Animals ; Cytoplasm/microbiology ; *Genome, Bacterial ; Insecta/genetics/*microbiology ; Interspersed Repetitive Sequences/genetics ; Reproduction/physiology ; Wolbachia/*genetics/*physiology ; }, abstract = {The effects have been considered that the intracellular symbiotic alpha-proteobacteria Wolbachia pipientis induces in its hosts, such as insects and other arthropods: cytoplasmic incompatibility upon mating, feminization, parthenogenesis, and androcide. Specific features of the bacterium genome and possible mechanisms of its action on hosts are discussed.}, } @article {pmid16202090, year = {2005}, author = {De Barro, PJ}, title = {Genetic structure of the whitefly Bemisia tabaci in the Asia-Pacific region revealed using microsatellite markers.}, journal = {Molecular ecology}, volume = {14}, number = {12}, pages = {3695-3718}, doi = {10.1111/j.1365-294X.2005.02700.x}, pmid = {16202090}, issn = {0962-1083}, mesh = {Animals ; Asia ; Australia ; Female ; *Genetic Variation ; Genotype ; Hemiptera/*genetics ; Indonesia ; Microsatellite Repeats ; *Phylogeny ; }, abstract = {Bemisia tabaci (Hemiptera: Aleyrodidae) is a haplo-diploid species of sap-feeding insect belonging to the group of insects commonly known as whiteflies. From earlier analyses of mitochondrial and ribosomal markers it has been concluded that in the Asia-Pacific region there were three major indigenous races as well as a large collection of genotypes with no clear association with any race. This new study uses 15 microsatellite loci and demonstrates that the indigenous Asia-Pacific genotypes can be split into six genetic populations with little or no gene flow between them. These bare only superficial similarity to the mitochondrial and ribosomal defined races. Moreover, four of the six can be further split into two subpopulations that again show little evidence gene flow between them. While the patterns reflect a strong geographical structure, physical barriers alone cannot explain all the observed structure. Differential host-plant utilization explained some of the substructure, but could not explain the overall structure. The roles of mating interference and Wolbachia in developing the genetic structure are considered. The lack of gene flow between genetic populations and some subpopulations further suggests that the barriers were either sufficiently impermeable to immigration or that reproductive isolation and competitive interactions were sufficiently strong to prevent gene flow. If the latter is the case, it suggests that there may be as many as 10 morphologically indistinguishable species indigenous to the Asia-Pacific region.}, } @article {pmid16198821, year = {2005}, author = {Simón, F and López-Belmonte, J and Marcos-Atxutegi, C and Morchón, R and Martín-Pacho, JR}, title = {What is happening outside North America regarding human dirofilariasis?.}, journal = {Veterinary parasitology}, volume = {133}, number = {2-3}, pages = {181-189}, doi = {10.1016/j.vetpar.2005.03.033}, pmid = {16198821}, issn = {0304-4017}, mesh = {Animals ; Antibodies, Bacterial/analysis/*blood ; Dirofilaria/*microbiology ; Dirofilaria immitis/microbiology ; Dirofilariasis/blood/*epidemiology/parasitology/transmission ; Europe/epidemiology ; Humans ; Sentinel Surveillance ; Seroepidemiologic Studies ; Symbiosis ; Wolbachia/*immunology ; }, abstract = {The etiologic agents of human dirofilariasis in the Old World are Dirofilaria immitis, which cause pulmonary and subcutaneous nodules, and Dirofilaria repens, which cause ocular lesions. Although reports of new cases of dirofilariasis are sporadic in other parts of the world, a considerable amount of information is generated in Europe regarding human dirofilariasis. Most cases have been detected in the Mediterranean countries, Ukraine, and Russia; however, isolated or short series of cases have been reported in the Balkan Republics and central and northern European countries. Seroepidemiologic studies have provided evidence that humans living in endemic areas present rates of infection similar to those of the autochthonous canine populations. Antibodies against endosymbiont Wolbachia bacteria have been demonstrated recently in human Dirofilaria infections. During D. immitis infections, preadult worms and third- and fourth-stage larvae are often destroyed by the host reaction, releasing a considerable amount of Wolbachia, and a Th1-type response against Wolbachia and/or filarial antigens is mounted. On the contrary, infections with D. repens, in which worms frequently remain intact, no Th1-type response has been observed. As humans are resistant hosts, the Th1-response could have a role in the resistance against parasites. The causes for the rise in the incidence of human dirofilariasis as well as the possible application of Wolbachia antigens in the serodiagnosis of human infections are discussed.}, } @article {pmid16198819, year = {2005}, author = {Kozek, WJ}, title = {What is new in the Wolbachia/Dirofilaria interaction?.}, journal = {Veterinary parasitology}, volume = {133}, number = {2-3}, pages = {127-132}, doi = {10.1016/j.vetpar.2005.02.005}, pmid = {16198819}, issn = {0304-4017}, support = {2 S06 GM8224/GM/NIGMS NIH HHS/United States ; AI 12095/AI/NIAID NIH HHS/United States ; RR-03051/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Antigens, Bacterial/analysis ; Cats ; Dirofilaria immitis/*microbiology ; Dirofilariasis/diagnosis/*parasitology ; Dogs ; Female ; Life Cycle Stages ; Male ; Symbiosis ; *Wolbachia/growth & development/immunology/pathogenicity/physiology ; }, abstract = {Presence of transovarially-transmitted endosymbiontic Wolbachia bacteria in Dirofilaria immitis, and in other filariae of man and animals, presents a new paradigm for our understanding of pathogenesis, treatment and diagnosis of filarial infections. Many of the basic biological characteristics of Wolbachia have yet to be elucidated, but the results obtained to date suggest that canine or the feline hosts can be exposed to D. immitis Wolbachia when larvae, or adult worms, are killed; when Wolbachia are expulsed, with the deposition of microfilariae, from the uterus of the females; and possibly through the excretory system of both male and female worms. The two organs that have the greatest potential of being affected by the Wolbachial metabolic products/antigens released from the adult worms are the lungs and the kidneys. Population of Wolbachia in D. immitis is polymorphic. The life cycle of Wolbachia is complex and may consist of two reproductive modes: multiplication of the bacillary forms by binary fission and by a more complex mode which resembles the Chlamydia-like cycle that consists of three morphological stages: a small, dense body, an intermediate stage with a dense inclusion, and a bacillary form which represents the final product of development and maturation of the small, dense body. The Chlamydia-like cycle offers a potential survival strategy for the Wolbachia by producing more progeny than multiplication by binary fission, and appears to be more active during growth and development of embryos and of the larvae. The small, dense bodies may be the infectious forms responsible for the spread of Wolbachia through the canalicular system, within the lateral chords of filariae. An amorphous membrane that lines the perienteric surface of the body wall may represent a physical barrier that limits the spread and movement of Wolbachia to the perienteric surface of the lateral chords. Wolbachia in D. immitis may also offer therapeutic and diagnostic possibilities. Elimination of Wolbachia by chemotherapy, and the suppressive effect of aposymbiosis on embryonic development of D. immitis, may have potential application for control (sterilization of female worms) and treatment of dirofilariasis. However, the three stages in the life cycle of Wolbachia may be antigenically different and each stage may have a different susceptibility to therapeutic agents. Persistence of dormant small, dense bodies after treatment would allow the Wolbachia to re-establish once the conditions for development would become favorable. Detection of Wolbachial antigens provides an attractive diagnostic possibility to identify D. immitis early in the infection. Further studies on Wolbachia of filariae, including those of D. immitis, will undoubtedly reveal additional information that can be applied towards treatment, diagnosis, and control of filarial infections.}, } @article {pmid16198306, year = {2005}, author = {Braquart-Varnier, C and Grève, P and Félix, C and Martin, G}, title = {Bacteriophage WO in Wolbachia infecting terrestrial isopods.}, journal = {Biochemical and biophysical research communications}, volume = {337}, number = {2}, pages = {580-585}, doi = {10.1016/j.bbrc.2005.09.091}, pmid = {16198306}, issn = {0006-291X}, mesh = {Animals ; Bacteriophages/*pathogenicity ; Base Sequence ; Feminization ; Isopoda/physiology/*virology ; Male ; Molecular Sequence Data ; Phenotype ; Wolbachia/*virology ; }, abstract = {Wolbachia are maternally inherited intracellular alpha-proteobacteria that infect a wide range of arthropods. They are associated with a number of different reproductive phenotypes in arthropods and nematodes. In isopod crustacean, Wolbachia are responsible for feminization of genetic males in many species, and for cytoplasmic incompatibility in two species. In this paper, we report the first detection of phage WO from Wolbachia infecting terrestrial isopods. All Wolbachia strains tested in this study were infected with phage WO. Based on the orf7 phage sequence, we identified three different phage sequences in four Wolbachia strains. The phage of Wolbachia infecting Armadillidium vulgare seems to be not active, unlike other phages WO previously described in arthropods.}, } @article {pmid16174339, year = {2005}, author = {Sakamoto, H and Ishikawa, Y and Sasaki, T and Kikuyama, S and Tatsuki, S and Hoshizaki, S}, title = {Transinfection reveals the crucial importance of Wolbachia genotypes in determining the type of reproductive alteration in the host.}, journal = {Genetical research}, volume = {85}, number = {3}, pages = {205-210}, doi = {10.1017/S0016672305007573}, pmid = {16174339}, mesh = {Animals ; Cytoplasm/genetics/physiology ; Female ; Genotype ; Host-Parasite Interactions/*physiology ; Lepidoptera/*genetics/microbiology ; Male ; Reproduction/genetics/*physiology ; Transfection/*methods ; Wolbachia/*genetics/physiology ; }, abstract = {Wolbachia , a group of endosymbiotic bacteria in arthropods, alter the reproduction of their hosts in various ways. A Wolbachia strain (wSca) naturally infecting the adzuki bean borer moth Ostrinia scapulalis induces male killing, while another strain (wKue) infecting the Mediterranean flour moth Ephestia kuehniella induces cytoplasmic incompatibility (CI) in the resident host. Transinfection of Wolbachia can be a powerful tool to elucidate the relative importance of Wolbachia and the host in determining the type of reproductive alterations. Recently, male killing was shown to occur in E. kuehniella transinfected with w Sca. In the present study, we transferred w Kue to O. scapulalis by embryonic microinjection. In the O. scapulalis transinfected with wKue, CI, but not male killing occurred. Thus, in addition to wSca, wKue was shown to induce the same type of alteration in a foreign host as in its natural host. These results demonstrate the crucial role of the Wolbachia genotype in determining the type of reproductive alteration. However, the present study also revealed the involvement of host factors. First, the degree of incompatibility induced by wKue in O. scapulalis was stronger than that in E. kuehniella , indicating that host factors can affect the level of CI. Second, the vertical transmission rate of wKue in O. scapulalis was generally low, suggesting that the host affects the dynamics of Wolbachia transmission.}, } @article {pmid16172008, year = {2005}, author = {Andrade, MA and Siles-Lucas, M and Pérez Arellano, JL and Pou Barreto, C and Valladares, B and Espinoza, E and Muro, A}, title = {Increased rat alveolar macrophage expression of functional iNOS induced by a Dirofilaria immitis immunoglobulin superfamily protein.}, journal = {Nitric oxide : biology and chemistry}, volume = {13}, number = {4}, pages = {217-225}, doi = {10.1016/j.niox.2005.06.003}, pmid = {16172008}, issn = {1089-8603}, mesh = {Animals ; Antigens/chemistry/metabolism ; Cells, Cultured ; Dirofilaria immitis/*immunology ; Immunoglobulins/chemistry/classification/*pharmacology ; Macrophages, Alveolar/*drug effects/*enzymology ; Male ; Nitric Oxide/metabolism ; Nitric Oxide Synthase Type II/*metabolism ; Rats ; Rats, Wistar ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; }, abstract = {Dirofilaria immitis is a worldwide filarial nematode causing heartworm disease in dogs and cats. Several mosquito species, which are able to feed both on humans and animals, can transmit this parasite. Inflammatory progression of host tissues induced by parasites are mediated by several molecules, including nitric oxide (NO), which usually exerts deleterious effects on parasites and occasionally on the host. We analyze the in vitro effect of total D. immitis adult worm somatic antigens on naïve rat alveolar macrophage NO production and further separation of parasite proteins to define specific D. immitis somatic molecules influencing host cell NO secretion. Additionally, we address the possible influence of Wolbachia spp. on the in vitro production of NO by macrophages. Our results demonstrate that D. immitis adult worm soluble antigens are able to specifically induce NO production from host macrophages. Furthermore, we demonstrated that this effect is due to nematode antigens rather than to defined components (LPS and metabolic molecules) derived from its endosymbiont, Wolbachia spp. In addition, we were able to isolate and identify one of the parasite specific components from the DiSo extract, denominated DiID35.3 and putatively belonging to the Immunoglobulin Superfamily Protein (ISP) group, triggering NO release from macrophages in a dose-dependent and specific manner.}, } @article {pmid16170768, year = {2005}, author = {Gilbert, J and Nfon, CK and Makepeace, BL and Njongmeta, LM and Hastings, IM and Pfarr, KM and Renz, A and Tanya, VN and Trees, AJ}, title = {Antibiotic chemotherapy of onchocerciasis: in a bovine model, killing of adult parasites requires a sustained depletion of endosymbiotic bacteria (Wolbachia species).}, journal = {The Journal of infectious diseases}, volume = {192}, number = {8}, pages = {1483-1493}, doi = {10.1086/462426}, pmid = {16170768}, issn = {0022-1899}, mesh = {Animals ; Cattle ; Cattle Diseases/drug therapy/immunology/*parasitology ; Disease Models, Animal ; Onchocerca/*microbiology ; Onchocerciasis/*drug therapy/*veterinary ; Symbiosis/*drug effects ; Tetracyclines/*pharmacology/therapeutic use ; Wolbachia/*drug effects/physiology ; }, abstract = {Development of a drug lethal to adult Onchocerca volvulus (i.e., macrofilaricide) is a research priority for the control of human onchocerciasis. Using bovine O. ochengi infections, we investigated the effects of oxytetracycline administered in a short intensive regimen (SIR; 10 mg/kg daily for 14 days), compared with a prolonged intermittent regimen (PIR; 20 mg/kg monthly for 6 months) or a combination of both (COM), on the viability of adult worms and their endosymbiotic bacteria (Wolbachia species). The long-term treatments eliminated >80% (COM) or >60% (PIR) of adult female worms (P<.001), and the COM regimen effected a sustained depletion of Wolbachia organisms. Conversely, SIR was not macrofilaricidal and only transiently depleted Wolbachia densities, which repopulated worm tissues by 24 weeks after treatment. These results unequivocally demonstrate the macrofilaricidal potential of tetracyclines against Onchocerca infection and suggest that intermittent, protracted administration will be more effective than continuous shorter term treatment.}, } @article {pmid16164550, year = {2005}, author = {Simser, JA and Rahman, MS and Dreher-Lesnick, SM and Azad, AF}, title = {A novel and naturally occurring transposon, ISRpe1 in the Rickettsia peacockii genome disrupting the rickA gene involved in actin-based motility.}, journal = {Molecular microbiology}, volume = {58}, number = {1}, pages = {71-79}, doi = {10.1111/j.1365-2958.2005.04806.x}, pmid = {16164550}, issn = {0950-382X}, support = {AI 17828/AI/NIAID NIH HHS/United States ; AI43006/AI/NIAID NIH HHS/United States ; }, mesh = {Actins/metabolism ; Amino Acid Sequence ; Antigens, Surface/genetics ; Bacterial Proteins/*genetics ; Base Sequence ; *DNA Transposable Elements ; DNA, Bacterial/chemistry/genetics ; Gene Rearrangement ; Genome, Bacterial ; Locomotion ; Molecular Sequence Data ; Phylogeny ; RNA, Bacterial/analysis ; RNA, Messenger/analysis ; Rickettsia/*genetics ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Terminal Repeat Sequences/genetics ; Transcription, Genetic ; Transposases/genetics ; Wolbachia/genetics ; }, abstract = {While examining the molecular basis for the lack of actin-based motility for the non-pathogenic spotted fever group (SFG) R. peacockii, we identified a novel insertion sequence (IS) element, ISRpe1, disrupting the coding sequence of rickA, demonstrated to induce actin-tail polymerization for the SFG rickettsiae. This rickettsial IS element appears to be active in that complete terminal inverted repeat and recombinase/transposase open reading frame sequences are present and the transposase is transcriptionally expressed. Phylogenetically, ISRpe1 belongs to a new IS family that is most closely related to those transposable elements of other intracellular bacteria like Wolbachia spp. ISRpe1 was demonstrated to be present in at least 10 locations throughout the R. peacockii genome, including one that disrupted the putative cell surface antigen encoding gene, sca1 considered to be involved in adhesion and virulence of the rickettsiae. Additionally, three IS sites demonstrated rearrangements/relocations of the R. peacockii genome when compared to those of other SFG rickettsiae. Our findings of the disruptions of rickA and sca1 along with the comparative genomic reassortments associated with ISRpe1 in the non-virulent R. peacockii provides opportunities to uncover molecular mechanisms underlying the pathogenesis and evolution of rickettsiae as well as its potential to be used in rickettsial transposon-based mutagenesis.}, } @article {pmid16156820, year = {2005}, author = {Charlat, S and Hornett, EA and Dyson, EA and Ho, PP and Loc, NT and Schilthuizen, M and Davies, N and Roderick, GK and Hurst, GD}, title = {Prevalence and penetrance variation of male-killing Wolbachia across Indo-Pacific populations of the butterfly Hypolimnas bolina.}, journal = {Molecular ecology}, volume = {14}, number = {11}, pages = {3525-3530}, doi = {10.1111/j.1365-294X.2005.02678.x}, pmid = {16156820}, issn = {0962-1083}, mesh = {Animals ; Asia, Southeastern ; Butterflies/*genetics/*microbiology ; DNA, Mitochondrial/genetics ; Female ; Genetic Variation ; Host-Parasite Interactions ; Male ; Pacific Islands ; Population Dynamics ; Sequence Analysis, DNA ; Sex Factors ; *Sex Ratio ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {Male-killing bacteria are generally thought to attain low to intermediate prevalence in natural populations, with only mild effects on the host population sex ratio. This view was recently challenged by reports of extremely high infection frequencies in three butterfly species, raising the prospect that male killers, by making males rare, might drive many features of host ecology and evolution. To assess this hypothesis, it is necessary to evaluate how often male killers actually produce a highly female-biased population sex ratio in nature, which requires both high prevalence of infection and high penetrance of action. To this end, we surveyed South Pacific and Southeast Asian populations of Hypolimnas bolina, a butterfly in which extreme prevalence of male-killing Wolbachia bacteria has recently been recorded. Our results indicate that highly female-biased populations are common in Polynesia, with 6 out of 12 populations studied having in excess of 70% of females infected with a fully efficient male killer. However, heterogeneity is extreme in Polynesia, with the male-killing Wolbachia absent from three populations. In contrast to the Polynesian situation, Wolbachia does not kill males in any of the three Southeast Asian populations studied, despite its very high prevalence there. We conclude that male killers are likely to have significant ongoing ecological and evolutionary impact in 6 of the 15 populations surveyed. The causes and consequences of the observed spatial variation are discussed with respect to host resistance evolution, host ecology and interference with additional symbionts.}, } @article {pmid16153037, year = {2005}, author = {Dyer, KA and Jaenike, J}, title = {Evolutionary dynamics of a spatially structured host-parasite association: Drosophila innubila and male-killing Wolbachia.}, journal = {Evolution; international journal of organic evolution}, volume = {59}, number = {7}, pages = {1518-1528}, pmid = {16153037}, issn = {0014-3820}, mesh = {Analysis of Variance ; Animals ; Arizona ; Bacterial Outer Membrane Proteins/genetics ; *Biological Evolution ; Cluster Analysis ; Drosophila/genetics/*microbiology ; Electron Transport Complex IV/genetics ; Founder Effect ; *Genetic Variation ; *Genetics, Population ; Geography ; Haplotypes/genetics ; Host-Parasite Interactions ; Population Dynamics ; Sequence Analysis, DNA ; Sex Factors ; *Sex Ratio ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {The mode and tempo of host-parasite evolution depend on population structure and history and the strength of selection that the species exert on each other. Here we genetically and epidemiologically characterize populations of the mycophagous fly Drosophila innubila and its male-killing Wolbachia endosymbiont, with the aim of integrating the local through global nature of this association. Drosophila innubila inhabit the forested "sky island" regions of the of the southwestern United States and northern Mexico, where its distribution is highly fragmented. We examine geographically isolated sky island populations of D. innubila, surveying the frequency and expression of Wolbachia infection as well as the distribution of genetic variation within and among populations of the host and parasite. In all populations, Wolbachia infection is associated with virtually complete male-killing, thus providing no evidence for the evolution of population-specific interaction phenotypes or local resistance. Although Wolbachia infection occurs in each of the main populations, there is variation among populations in the prevalence of infection and the resulting population-level sex ratio of D. innubila. Among these populations, the nuclear genes of D. innubila show moderate, though significant, differentiation. In contrast, the host mitochondrial DNA (mtDNA), which shares transmission with Wolbachia, exhibits substantially greater geographic differentiation, even after accounting for differences in transmission between nuclear and mitochondrial genes. We suggest that this pattern is caused by local Wolbachia--but not D. innubila--fluctuations in prevalence that increase the severity of drift experienced only by the mtDNA. Overall, our data suggest that the association between D. innubila and male-killing Wolbachia is ecologically dynamic within local populations, but evolutionarily coherent across the species as a whole.}, } @article {pmid16132739, year = {2005}, author = {Enigl, M and Zchori-Fein, E and Schausberger, P}, title = {Negative evidence of Wolbachia in the predaceous mite Phytoseiulus persimilis.}, journal = {Experimental & applied acarology}, volume = {36}, number = {4}, pages = {249-262}, pmid = {16132739}, issn = {0168-8162}, mesh = {Africa ; Animals ; Australia ; Europe ; Female ; Male ; Mites/microbiology/*physiology ; Ovum ; *Predatory Behavior ; Reproduction ; Time Factors ; United States ; Wolbachia/*isolation & purification ; }, abstract = {The cytoplasmically inherited bacterium Wolbachia is widespread in arthropod species and has been repeatedly detected in the predaceous mite Phytoseiulus persimilis. Our original goal was to assess the prevalence of Wolbachia infection in P. persimilis and the potential fitness consequences for this host. To accomplish that goal, seven P. persimilis strains were obtained from Europe, Africa and the USA and reared on the phytophagous mite Tetranychus urticae as prey. After preliminary results showed that the T. urticae used was infected with Wolbachia, the minimum starvation time of the predators to prevent false positive results from undigested prey was determined. We tested DNA samples by PCR (polymerase chain reaction) after starving the predators or feeding them Wolbachia-free T. urticae for various periods. Those experiments showed that Wolbachia could not be detected after 16 h at 25 degrees C and 48 h at 20 degrees C. To verify the results of the PCR analyses, we furthermore conducted crossing experiments with antibiotic-treated and untreated individuals. No indications of Wolbachia effects were recorded. Additionally, we screened live eggs of four of the seven strains reared in our laboratory and alcohol samples of 10 other P. persimilis strains for the occurrence of Wolbachia by PCR, none of which tested positive. Synthesis of our study and previous reports suggests that infection of P. persimilis with Wolbachia is extremely rare and of minor importance. We discuss the significance of our findings for future studies on the presence of Wolbachia in predaceous arthropods.}, } @article {pmid16118660, year = {2005}, author = {Dedeine, F and Boulétreau, M and Vavre, F}, title = {Wolbachia requirement for oogenesis: occurrence within the genus Asobara (Hymenoptera, Braconidae) and evidence for intraspecific variation in A. tabida.}, journal = {Heredity}, volume = {95}, number = {5}, pages = {394-400}, doi = {10.1038/sj.hdy.6800739}, pmid = {16118660}, issn = {0018-067X}, mesh = {Animals ; Drosophila/genetics ; Female ; Fertility ; Host-Parasite Interactions/genetics/physiology ; Hymenoptera/genetics/*physiology ; Larva/growth & development ; Oocytes/cytology/enzymology ; Oogenesis/*physiology ; Species Specificity ; Symbiosis/genetics/physiology ; Wasps/genetics/*physiology ; Wolbachia/*genetics/pathogenicity/physiology ; }, abstract = {Wolbachia are symbiotic bacteria that induce a diversity of phenotypes on their numerous invertebrate host species. In the wasp Asobara tabida (Braconidae), each individual harbours three Wolbachia strains: wAtab3, which is required for host oogenesis, and wAtab1 and wAtab2, that do not have this function but induce cytoplasmic incompatibility. In this study, we surveyed and identified Wolbachia strains in four additional Asobara species. We detected Wolbachia in one of these species, but both the identity (based on wsp gene) and prevalence of the Wolbachia detected in natural population indicate that this host species is not dependent on Wolbachia for oogenesis. We also compared A. tabida lines of different geographical origin for their dependence on Wolbachia. All individuals from 16 A. tabida lines proved to be infected by the three Wolbachia strains wAtab1, wAtab2 and wAtab3, but, interestingly, we found variation among lines in the degree to which females were dependent on Wolbachia to produce their oocytes. In three lines, aposymbiotic females (cured from the three Wolbachia strains by antibiotics) can produce some oocytes. However, these aposymbiotic females produce fewer and smaller oocytes than symbiotic ones, and the larvae they produce die before full development. Thus, depending on which nuclear genotype they have, A. tabida females depend on Wolbachia either because they fail to produce any oocyte or because the few oocytes they do produce generate unviable offspring. We discuss the implications of these findings for the understanding of the physiological and genetic deficiency of aposymbiotic females.}, } @article {pmid16118145, year = {2002}, author = {Gong, P and Shen, ZR}, title = {[Molecular diagnostic techniques of Wolbachia].}, journal = {Yi chuan = Hereditas}, volume = {24}, number = {2}, pages = {207-210}, pmid = {16118145}, issn = {0253-9772}, abstract = {Wolbachia is a common and widespread group of bacteria found in reproductive tissues of arthropods. These bacteria are transmitted through the cytoplasm of eggs and have evolved various mechanisms for manipulating reproduction of their hosts. Because of important relationship with some major biological issues and implication of biological control and genetic engineering, the study of Wolbachia has under gone an explosive growth in recent years. A tremendous progress has been made based on the advancing molecular diagnostic technique of Wolbachia. In this paper, those molecular diagnostic techniques are detailed and it is aimed to be the primer for the research of Wolbachia in China.}, } @article {pmid16106260, year = {2005}, author = {Sasaki, T and Massaki, N and Kubo, T}, title = {Wolbachia variant that induces two distinct reproductive phenotypes in different hosts.}, journal = {Heredity}, volume = {95}, number = {5}, pages = {389-393}, doi = {10.1038/sj.hdy.6800737}, pmid = {16106260}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Female ; *Genetic Variation ; Gram-Negative Bacterial Infections ; Host-Parasite Interactions/*genetics ; Male ; Moths/genetics/parasitology ; Ovary/parasitology ; Phenotype ; Polymerase Chain Reaction ; Symbiosis/genetics ; Tetracycline/pharmacology ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia is an intracellular endosymbiont that induces a variety of reproductive alterations in diverse arthropods. The almond moth, Cadra cautella, is double infected with two Wolbachia variants, wCauA and wCauB, and expresses complete cytoplasmic incompatibility (CI). The individual contribution of wCauA and wCauB to the expression of CI are unclear, however, because the two variants have not been separated in this host. The effect of wCauA is of particular interest because it induces male killing when transferred into the Mediterranean flour moth, Ephestia kuehniella. In the present study, we generated C. cautella infected with only wCauA by treating double-infected insects with tetracycline. Single-infected C. cautella exhibited strong CI, demonstrating that wCauA induces two distinct reproductive phenotypes in different hosts: CI in C. cautella and male killing in E. kuehniella. CI was also observed in the cross of double-infected males and single-infected females. Comparison of the single- and double-infected insects by real-time quantitative polymerase chain reaction suggested that the wCauA density is not affected much by the presence or absence of wCauB.}, } @article {pmid16103603, year = {2005}, author = {Plichart, C and Legrand, AM}, title = {Detection and characterization of Wolbachia infections in Wuchereria bancrofti (Spirurida: Onchocercidae) var. pacifica and Aedes (Stegomyia) polynesiensis (Diptera: Culicidae).}, journal = {The American journal of tropical medicine and hygiene}, volume = {73}, number = {2}, pages = {354-358}, pmid = {16103603}, issn = {0002-9637}, mesh = {Aedes/*microbiology/parasitology ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; Filariasis/parasitology/transmission ; Microfilariae/microbiology ; Molecular Sequence Data ; Pest Control, Biological ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/classification/genetics/growth & development/*isolation & purification ; Wuchereria bancrofti/*microbiology ; }, abstract = {Despite control programs based on mass drug administration (MDA) of microfilaricidal compounds, Bancroftian lymphatic filariasis remains a problem in French Polynesia. For an alternative strategy to MDA, we investigated the potential role of Wolbachia to control filarial transmission. Wolbachia are intracellular alpha-proteobacteria endosymbionts that infect a broad range of insects and nematodes. These bacteria have a suspected role in the pathogenesis of filariasis. They also may be useful in mosquito control through cytoplasmic incompatibility. To detect and characterize these bacteria in the filarial and mosquito-vectors in French Polynesia, a survey was conducted on field-collected mosquitoes and microfilariae from infected people. Samples were analyzed by a polymerase chain reaction and gene sequencing. The results indicate that these bacteria are widespread. Sequence analysis of the wsp and ftsZ genes positioned the Aedes polynesiensis Wolbachia in cluster A and Wuchereria bancrofti var. pacifica Wolbachia in cluster D. The implications for possible improved treatment and vector control are discussed.}, } @article {pmid16103005, year = {2005}, author = {Gunawardena, NK and Fujimaki, Y and Aoki, Y and Mishima, N and Ezaki, T and Uni, S and Kimura, E}, title = {Differential effects of diethylcarbamazine, tetracycline and the combination on Brugia pahangi adult females in vitro.}, journal = {Parasitology international}, volume = {54}, number = {4}, pages = {253-259}, doi = {10.1016/j.parint.2005.06.005}, pmid = {16103005}, issn = {1383-5769}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/*pharmacology ; Brugia pahangi/*drug effects/embryology/microbiology/physiology ; Culture Media ; Diethylcarbamazine/administration & dosage/*pharmacology ; Drug Therapy, Combination ; Female ; Filaricides/administration & dosage/*pharmacology ; Microfilariae/drug effects/microbiology ; Parasitic Sensitivity Tests ; Symbiosis ; Tetracycline/administration & dosage/*pharmacology ; Wolbachia/drug effects ; }, abstract = {Anti-filarial effects of diethylcarbamazine (DEC), tetracycline (TC) and the combination on Brugia pahangi adult females were studied in 7-day cell-free culture, in terms of microfilaria release, parasite motility, MTT assay for parasite viability and embryogram. TC 50 microg/ml (TC50) effectively reduced microfilaria release from day 1 of culture. Combined with DEC 100 microg/ml (DEC100) or DEC 500 microg/ml (DEC500), microfilaria release reduced further and synergistically. TC50 also reduced motility, but DEC100 and DEC500 did not. The combination of TC50 and DEC500 reduced motility synergistically. The MTT assay supported the results of motility study in general. The embryogram showed that only DEC500 reduced the total number of intrauterine embryos, especially ova, indicating that DEC500 inhibited early embryogenesis. TC50 did not affect the total number of embryos, but resulted in apparent accumulation of microfilariae in the uterus, suggesting that the drug inhibited release of microfilariae in this in vitro system. These results clarified different anti-female mechanisms between DEC and TC. A PCR-based study showed that endosymbiont bacteria, Wolbachia, in B. pahangi females decreased significantly after TC treatment. However, this study could not determine whether the effects of TC were direct or Wolbachia-mediated.}, } @article {pmid16085497, year = {2005}, author = {Riegler, M and Sidhu, M and Miller, WJ and O'Neill, SL}, title = {Evidence for a global Wolbachia replacement in Drosophila melanogaster.}, journal = {Current biology : CB}, volume = {15}, number = {15}, pages = {1428-1433}, doi = {10.1016/j.cub.2005.06.069}, pmid = {16085497}, issn = {0960-9822}, mesh = {Animals ; Chromosome Mapping ; Chromosomes, Bacterial/*genetics ; DNA Primers ; Drosophila melanogaster/*microbiology ; Genetic Markers/genetics ; Genotype ; Minisatellite Repeats/genetics ; *Polymorphism, Genetic ; Reproduction/genetics ; Sequence Analysis, DNA ; Species Specificity ; Wolbachia/*genetics ; }, abstract = {Wolbachia are maternally inherited intracellular alpha-Proteobacteria found in numerous arthropod and filarial nematode species. They influence the biology of their hosts in many ways. In some cases, they act as obligate mutualists and are required for the normal development and reproduction of the host. They are best known, however, for the various reproductive parasitism traits that they can generate in infected hosts. These include cytoplasmic incompatibility (CI) between individuals of different infection status, the parthenogenetic production of females, the selective killing of male embryos, and the feminization of genetic males. Wolbachia infections of Drosophila melanogaster are extremely common in both wild populations and long-term laboratory stocks. Utilizing the newly completed genome sequence of Wolbachia pipientis wMel, we have identified a number of polymorphic markers that can be used to discriminate among five different Wolbachia variants within what was previously thought to be the single clonal infection of D. melanogaster. Analysis of long-term lab stocks together with wild-caught flies indicates that one of these variants has replaced the others globally within the last century. This is the first report of a global replacement of a Wolbachia strain in an insect host species. The sweep is at odds with current theory that cannot explain how Wolbachia can invade this host species given the observed cytoplasmic incompatibility characteristics of Wolbachia infections in D. melanogaster in the field.}, } @article {pmid16033575, year = {2005}, author = {Pannebakker, BA and Schidlo, NS and Boskamp, GJ and Dekker, L and van Dooren, TJ and Beukeboom, LW and Zwaan, BJ and Brakefield, PM and van Alphen, JJ}, title = {Sexual functionality of Leptopilina clavipes (Hymenoptera: Figitidae) after reversing Wolbachia-induced parthenogenesis.}, journal = {Journal of evolutionary biology}, volume = {18}, number = {4}, pages = {1019-1028}, doi = {10.1111/j.1420-9101.2005.00898.x}, pmid = {16033575}, issn = {1010-061X}, mesh = {*Alleles ; Animals ; Crosses, Genetic ; Europe ; Female ; Fertility/physiology ; Male ; Parthenogenesis/*physiology ; Polymorphism, Restriction Fragment Length ; Rifampin ; Sex Ratio ; Sexual Behavior, Animal/*physiology ; Spermatozoa/physiology ; Wasps/genetics/*microbiology/*physiology ; *Wolbachia ; }, abstract = {Females infected with parthenogenesis-inducing Wolbachia bacteria can be cured from their infection by antibiotic treatment, resulting in male production. In most cases, however, these males are either sexually not fully functional, or infected females have lost the ability to reproduce sexually. We studied the decay of sexual function in males and females of the parasitoid Leptopilina clavipes. In western Europe, infected and uninfected populations occur allopatrically, allowing for an investigation of both male and female sexual function. This was made by comparing females and males induced from different parthenogenetic populations with those from naturally occurring uninfected populations. Our results indicate that although males show a decay of sexual function, they are still able to fertilize uninfected females. Infected females, however, do not fertilize their eggs after mating with males from uninfected populations. The absence of genomic incompatibilities suggests that these effects are due to the difference in mode of reproduction.}, } @article {pmid16033569, year = {2005}, author = {Champion de Crespigny, FE and Butlin, RK and Wedell, N}, title = {Can cytoplasmic incompatibility inducing Wolbachia promote the evolution of mate preferences?.}, journal = {Journal of evolutionary biology}, volume = {18}, number = {4}, pages = {967-977}, doi = {10.1111/j.1420-9101.2005.00909.x}, pmid = {16033569}, issn = {1010-061X}, mesh = {Animals ; *Biological Evolution ; Computer Simulation ; Female ; Gene Frequency ; *Genetics, Population ; Host-Parasite Interactions ; Insecta/*microbiology ; Male ; *Models, Biological ; Population Dynamics ; Reproduction/physiology ; Sexual Behavior, Animal/*physiology ; *Wolbachia ; }, abstract = {The maternally inherited bacterium, Wolbachia pipientis, manipulates host reproduction by rendering uninfected females reproductively incompatible with infected males (cytoplasmic incompatibility, CI). Hosts may evolve mechanisms, such as mate preferences, to avoid fitness costs of Wolbachia infection. Despite the potential importance of mate choice for Wolbachia population dynamics, this possibility remains largely unexplored. Here we model the spread of an allele encoding female mate preference for uninfected males alongside the spread of CI inducing Wolbachia. Mate preferences can evolve but the spread of the preference allele depends on factors associated with both Wolbachia infection and the preference allele itself. Incomplete maternal transmission of Wolbachia, fitness costs and low CI, improve the spread of the preference allele and impact on the population dynamics of Wolbachia. In addition, mate preferences are found in infected individuals. These results have important consequences for the fate of Wolbachia and studies addressing mate preferences in infected populations.}, } @article {pmid16030235, year = {2005}, author = {Baldo, L and Lo, N and Werren, JH}, title = {Mosaic nature of the wolbachia surface protein.}, journal = {Journal of bacteriology}, volume = {187}, number = {15}, pages = {5406-5418}, pmid = {16030235}, issn = {0021-9193}, mesh = {Amino Acid Sequence ; Bacterial Outer Membrane Proteins/*genetics ; Molecular Sequence Data ; Phylogeny ; *Recombination, Genetic ; Sequence Alignment ; Wolbachia/*genetics ; }, abstract = {Lateral gene transfer and recombination play important roles in the evolution of many parasitic bacteria. Here we investigate intragenic recombination in Wolbachia bacteria, considered among the most abundant intracellular bacteria on earth. We conduct a detailed analysis of the patterns of variation and recombination within the Wolbachia surface protein, utilizing an extensive set of published and new sequences from five main supergroups of Wolbachia. Analysis of nucleotide and amino acid sequence variations confirms four hypervariable regions (HVRs), separated by regions under strong conservation. Comparison of shared polymorphisms reveals a complex mosaic structure of the gene, characterized by a clear intragenic recombining of segments among several distinct strains, whose major recombination effect is shuffling of a relatively conserved set of amino acid motifs within each of the four HVRs. Exchanges occurred both within and between the arthropod supergroups. Analyses based on phylogenetic methods and a specific recombination detection program (MAXCHI) significantly support this complex partitioning of the gene, indicating a chimeric origin of wsp. Although wsp has been widely used to define macro- and microtaxonomy among Wolbachia strains, these results clearly show that it is not suitable for this purpose. The role of wsp in bacterium-host interactions is currently unknown, but results presented here indicate that exchanges of HVR motifs are favored by natural selection. Identifying host proteins that interact with wsp variants should help reveal how these widespread bacterial parasites affect and evolve in response to the cellular environments of their invertebrate hosts.}, } @article {pmid16030207, year = {2005}, author = {Iturbe-Ormaetxe, I and Burke, GR and Riegler, M and O'Neill, SL}, title = {Distribution, expression, and motif variability of ankyrin domain genes in Wolbachia pipientis.}, journal = {Journal of bacteriology}, volume = {187}, number = {15}, pages = {5136-5145}, pmid = {16030207}, issn = {0021-9193}, mesh = {Ankyrins/*genetics/metabolism ; Genetic Variation ; *Genome, Bacterial ; Molecular Sequence Data ; Protein Structure, Tertiary/genetics ; RNA, Messenger/genetics ; Wolbachia/*genetics ; }, abstract = {The endosymbiotic bacterium Wolbachia pipientis infects a wide range of arthropods, in which it induces a variety of reproductive phenotypes, including cytoplasmic incompatibility (CI), parthenogenesis, male killing, and reversal of genetic sex determination. The recent sequencing and annotation of the first Wolbachia genome revealed an unusually high number of genes encoding ankyrin domain (ANK) repeats. These ANK genes are likely to be important in mediating the Wolbachia-host interaction. In this work we determined the distribution and expression of the different ANK genes found in the sequenced Wolbachia wMel genome in nine Wolbachia strains that induce different phenotypic effects in their hosts. A comparison of the ANK genes of wMel and the non-CI-inducing wAu Wolbachia strain revealed significant differences between the strains. This was reflected in sequence variability in shared genes that could result in alterations in the encoded proteins, such as motif deletions, amino acid insertions, and in some cases disruptions due to insertion of transposable elements and premature stops. In addition, one wMel ANK gene, which is part of an operon, was absent in the wAu genome. These variations are likely to affect the affinity, function, and cellular location of the predicted proteins encoded by these genes.}, } @article {pmid16015330, year = {2005}, author = {Sinkins, SP and Walker, T and Lynd, AR and Steven, AR and Makepeace, BL and Godfray, HC and Parkhill, J}, title = {Wolbachia variability and host effects on crossing type in Culex mosquitoes.}, journal = {Nature}, volume = {436}, number = {7048}, pages = {257-260}, doi = {10.1038/nature03629}, pmid = {16015330}, issn = {1476-4687}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Amino Acid Sequence ; Animals ; Bacterial Infections/microbiology/physiopathology ; Cell Nucleus/genetics ; Crosses, Genetic ; Culex/classification/genetics/*microbiology/*physiology ; Cytoplasm/*physiology ; Female ; Genes, Bacterial/genetics ; Genetic Variation/*genetics ; Genome ; Host-Parasite Interactions ; Male ; Molecular Sequence Data ; Prophages/genetics ; RNA, Messenger/genetics/metabolism ; Reproduction/genetics/physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Wolbachia/classification/*genetics/*physiology ; }, abstract = {Wolbachia is a common maternally inherited bacterial symbiont able to induce crossing sterilities known as cytoplasmic incompatibility (CI) in insects. Wolbachia-modified sperm are unable to complete fertilization of uninfected ova, but a rescue function allows infected eggs to develop normally. By providing a reproductive advantage to infected females, Wolbachia can rapidly invade uninfected populations, and this could provide a mechanism for driving transgenes through pest populations. CI can also occur between Wolbachia-infected populations and is usually associated with the presence of different Wolbachia strains. In the Culex pipiens mosquito group (including the filariasis vector C. quinquefasciatus) a very unusual degree of complexity of Wolbachia-induced crossing-types has been reported, with partial or complete CI that can be unidirectional or bidirectional, yet no Wolbachia strain variation was found. Here we show variation between incompatible Culex strains in two Wolbachia ankyrin repeat-encoding genes associated with a prophage region, one of which is sex-specifically expressed in some strains, and also a direct effect of the host nuclear genome on CI rescue.}, } @article {pmid16015317, year = {2005}, author = {Hoffmann, AA}, title = {Entomology: incompatible mosquitoes.}, journal = {Nature}, volume = {436}, number = {7048}, pages = {189}, doi = {10.1038/436189a}, pmid = {16015317}, issn = {1476-4687}, mesh = {Animals ; Bacterial Infections/microbiology/physiopathology ; Crosses, Genetic ; Culex/cytology/genetics/*microbiology/*physiology ; Cytoplasm/*physiology ; Female ; Host-Parasite Interactions ; Male ; Mosquito Control ; Reproduction/genetics/physiology ; Wolbachia/classification/genetics/*physiology ; }, } @article {pmid16005284, year = {2005}, author = {Brownlie, JC and O'Neill, SL}, title = {Wolbachia genomes: insights into an intracellular lifestyle.}, journal = {Current biology : CB}, volume = {15}, number = {13}, pages = {R507-9}, doi = {10.1016/j.cub.2005.06.029}, pmid = {16005284}, issn = {0960-9822}, mesh = {Animals ; Brugia malayi/genetics/*microbiology ; *Genome, Bacterial ; Heme/genetics ; *Phenotype ; Repetitive Sequences, Nucleic Acid/genetics ; Riboflavin/genetics ; Species Specificity ; Symbiosis/*genetics ; Wolbachia/*genetics/physiology ; }, abstract = {The genome sequence of the Wolbachia endosymbiont that infects the nematode Brugia malayi has recently been determined together with three partial Wolbachia genomes from different Drosophila species. These data along with the previously published Wolbachia genome from Drosophila melanogaster provide new insights into how this endosymbiont has managed to become so successful.}, } @article {pmid16001025, year = {2005}, author = {Ainsworth, C}, title = {Parasite infiltrates fruitfly research.}, journal = {Nature}, volume = {436}, number = {7047}, pages = {8}, doi = {10.1038/436008a}, pmid = {16001025}, issn = {1476-4687}, mesh = {Animals ; Artifacts ; Drosophila melanogaster/genetics/*parasitology/*physiology ; Female ; Host-Parasite Interactions ; Male ; Parasites/genetics/isolation & purification/*physiology ; Research/*standards ; Symbiosis ; Wolbachia/genetics/isolation & purification/*physiology ; }, } @article {pmid15998463, year = {2005}, author = {Salzberg, SL and Dunning Hotopp, JC and Delcher, AL and Pop, M and Smith, DR and Eisen, MB and Nelson, WC}, title = {Correction: Serendipitous discovery of Wolbachia genomes in multiple Drosophila species.}, journal = {Genome biology}, volume = {6}, number = {7}, pages = {402}, pmid = {15998463}, issn = {1474-760X}, support = {R01 LM006845/LM/NLM NIH HHS/United States ; }, mesh = {Animals ; Drosophila/genetics/*microbiology ; *Genome, Bacterial ; Wolbachia/*genetics/isolation & purification ; }, abstract = {A correction to Serendipitous discovery of Wolbachia genomes in multiple Drosophila species by SL Salzberg, JC Dunning Hotopp, AL Delcher, M Pop, DR Smith, MB Eisen and WC Nelson. Genome Biology 2005, 6:R23.}, } @article {pmid15998462, year = {2005}, author = {Iturbe-Ormaetxe, I and Riegler, M and O'Neill, SL}, title = {New names for old strains? Wolbachia wSim is actually wRi.}, journal = {Genome biology}, volume = {6}, number = {7}, pages = {401; author reply 401}, pmid = {15998462}, issn = {1474-760X}, mesh = {Animals ; Drosophila/microbiology ; Gene Transfer, Horizontal ; Genome ; Genomics ; Phylogeny ; Symbiosis ; Wolbachia/*classification/*genetics/physiology ; }, abstract = {A response to Serendipitous discovery of Wolbachia genomes in multiple Drosophila species by SL Salzberg, JC Dunning Hotopp, AL Delcher, M Pop, DR Smith, MB Eisen and WC Nelson. Genome Biology 2005, 6:R23}, } @article {pmid15997410, year = {2005}, author = {Müller, HA and Zahner, H}, title = {Lethal LPS-independent side effects after microfilaricidal treatment in Acanthocheilonema viteae-infected rodents.}, journal = {Parasitology research}, volume = {97}, number = {3}, pages = {201-208}, pmid = {15997410}, issn = {0932-0113}, mesh = {Animals ; Diethylcarbamazine/*adverse effects ; *Dipetalonema/immunology/isolation & purification ; Dipetalonema Infections/*drug therapy/mortality/parasitology ; Disease Models, Animal ; Drug Therapy, Combination ; Female ; Filaricides/*adverse effects ; Gerbillinae ; Levamisole/*adverse effects ; *Lipopolysaccharides/metabolism ; Microfilariae/drug effects ; Muridae ; Parasitic Diseases, Animal/*drug therapy/mortality ; Rodent Diseases/drug therapy/parasitology ; Species Specificity ; Survival Rate ; Wolbachia/immunology ; }, abstract = {Mastomys coucha and jirds infected with Acanthocheilonema viteae, a filarial species free of endosymbiontic bacteria of the genus Wolbachia, suffer lethal side effects after effective microfilaricidal therapy with diethylcarbamazine and levamisole, whereas, M. coucha infected with the Wolbachia-infested species Brugia malayi or Litomosoides carinii tolerate corresponding treatment. Mortality in A. viteae infected, treated animals varied with microfilariae density in the blood. It was up to 100% in highly microfilaraemic M. coucha and jirds, but low or absent in animals with low microfilariae counts. Deaths occurred in most cases 5-24 h after treatment. Characteristic symptoms in animals, which died subsequently were a rapid drop in body temperature by 4-7 degrees C, an increase in hematokrit values by up to 10% and a moderate blood acidosis. Lethal effects in A. viteae infections did not depend on a particular status of hypersensitivity of the animals since desensitization procedures, which protected infected M. coucha against an otherwise lethal intravenous challenge with A. viteae homogenate did not protect against adverse reactions to a subsequent microfilaricidal treatment. The animals were protected from treatment induced death by injection of N-LMMA. Thus the final morbific agent seems NO. The data show that adverse effects after effective microfilaricidal therapy may be caused by microfilariae derived components different from Wolbachia-released LPS.}, } @article {pmid15997407, year = {2005}, author = {Rao, R and Moussa, H and Vanderwaal, RP and Sampson, E and Atkinson, LJ and Weil, GJ}, title = {Effects of gamma radiation on Brugia malayi infective larvae and their intracellular Wolbachia bacteria.}, journal = {Parasitology research}, volume = {97}, number = {3}, pages = {219-227}, pmid = {15997407}, issn = {0932-0113}, mesh = {Animals ; Antibodies, Bacterial/immunology ; Bacterial Outer Membrane Proteins/immunology ; Brugia malayi/microbiology/*radiation effects ; DNA, Bacterial/radiation effects ; Dose-Response Relationship, Radiation ; Filariasis/immunology/prevention & control/*radiotherapy ; Gamma Rays/*adverse effects/*therapeutic use ; Gerbillinae ; Host-Parasite Interactions/immunology/*radiation effects ; Larva/microbiology/radiation effects ; Parasitemia/diagnosis/immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccination ; Wolbachia/genetics/isolation & purification/*radiation effects/ultrastructure ; }, abstract = {Prior studies have shown that irradiated filarial larvae are developmentally stunted but capable of inducing partial immunity to filariasis in animals. The mechanisms for these effects are poorly understood. Recent studies suggest that intracellular Wolbachia bacteria are necessary for the normal development, reproduction and survival of filarial nematodes. The purpose of this study was to examine the effects of irradiation on Wolbachia in Brugia malayi infective larvae (L3) and on L3 development. The L3 were exposed to 0, 25, 35, 45, 55, 65 or 75 krad of gamma irradiation from a (137) Cesium source and cultured in vitro at 37 degrees C in NCTC/IMDM medium with 10% FCS for 12 days. Irradiation prevented molting of L3 to the L4 stage in a dose-dependent manner. Electron microscopy studies showed that irradiation damaged Wolbachia (25 krad) or cleared them from worm tissues (45 krad). In addition, majority of the irradiated L3s failed to develop the L4 cuticle. Real-time PCR studies showed that irradiation reduced Wolbachia DNA in worm tissues. Parallel in vivo studies confirmed decreased development of irradiated L3 in jirds, with associated effects on Wolbachia. Jirds injected s.c with normal L3 developed antibodies to Wolbachia surface protein (wsp) shortly after the onset of microfilarial patency. In contrast, jirds injected with irradiated L3 did not develop microfilaremia or antibodies to wsp. Additional studies are needed to test the hypothesis that irradiation retards growth and development of filarial L3 by killing Wolbachia.}, } @article {pmid15964448, year = {2005}, author = {Taylor, MJ and Makunde, WH and McGarry, HF and Turner, JD and Mand, S and Hoerauf, A}, title = {Macrofilaricidal activity after doxycycline treatment of Wuchereria bancrofti: a double-blind, randomised placebo-controlled trial.}, journal = {Lancet (London, England)}, volume = {365}, number = {9477}, pages = {2116-2121}, doi = {10.1016/S0140-6736(05)66591-9}, pmid = {15964448}, issn = {1474-547X}, mesh = {Adolescent ; Adult ; Aged ; Animals ; Anti-Bacterial Agents/*therapeutic use ; Double-Blind Method ; Doxycycline/*therapeutic use ; Elephantiasis, Filarial/*drug therapy/parasitology ; Humans ; Male ; Middle Aged ; Wolbachia/*drug effects ; Wuchereria bancrofti/drug effects/*microbiology ; }, abstract = {BACKGROUND: Wolbachia endosymbionts of filarial nematodes are vital for larval development and adult-worm fertility and viability. This essential dependency on the bacterium for survival of the parasites has provided a new approach to treat filariasis with antibiotics. We used this strategy to investigate the effects of doxycycline treatment on the major cause of lymphatic filariasis, Wuchereria bancrofti.

METHODS: We undertook a double-blind, randomised, placebo-controlled field trial of doxycycline (200 mg per day) for 8 weeks in 72 individuals infected with W bancrofti from Kimang'a village, Pangani, Tanzania. Participants were randomly assigned by block randomisation to receive capsules of doxycycline (n=34) or placebo (n=38). We assessed treatment efficacy by monitoring microfilaraemia, antigenaemia, and ultrasound detection of adult worms. Follow-up assessments were done at 5, 8, 11, and 14 months after the start of treatment. Analysis was per protocol.

FINDINGS: One person from the doxycycline group died from HIV infection. Five (doxycycline) and 11 (placebo) individuals were absent at the time of ultrasound analysis. Doxycycline treatment almost completely eliminated microfilaraemia at 8-14 months' follow-up (for all timepoints p<0.001). Ultrasonography detected adult worms in only six (22%) of 27 individuals treated with doxycycline compared with 24 (88%) of 27 with placebo at 14 months after the start of treatment (p<0.0001). At the same timepoint, filarial antigenaemia in the doxycycline group fell to about half of that before treatment (p=0.015). Adverse events were few and mild.

INTERPRETATION: An 8-week course of doxycycline is a safe and well-tolerated treatment for lymphatic filariasis with significant activity against adult worms and microfilaraemia.}, } @article {pmid15964428, year = {2005}, author = {Stolk, WA and de Vlas, SJ and Habbema, JD}, title = {Anti-Wolbachia treatment for lymphatic filariasis.}, journal = {Lancet (London, England)}, volume = {365}, number = {9477}, pages = {2067-2068}, doi = {10.1016/S0140-6736(05)66714-1}, pmid = {15964428}, issn = {1474-547X}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Elephantiasis, Filarial/*drug therapy/parasitology ; Filaricides/therapeutic use ; Humans ; Wolbachia/*drug effects ; Wuchereria bancrofti/drug effects/*microbiology ; }, } @article {pmid15955513, year = {2005}, author = {Dean, MD and Ballard, JW}, title = {High divergence among Drosophila simulans mitochondrial haplogroups arose in midst of long term purifying selection.}, journal = {Molecular phylogenetics and evolution}, volume = {36}, number = {2}, pages = {328-337}, doi = {10.1016/j.ympev.2005.02.016}, pmid = {15955513}, issn = {1055-7903}, mesh = {Animals ; Codon/genetics ; DNA, Mitochondrial/*genetics ; Drosophila/classification/*genetics ; *Genetic Variation ; Haplotypes/*genetics ; Models, Genetic ; Phylogeny ; Point Mutation ; *Selection, Genetic ; Species Specificity ; }, abstract = {We characterize the type of selection acting within and among mitochondrial lineages in five closely related Drosophila species. We focus on D. simulans, where three genetically distinct mitochondrial haplogroups show high interhaplogroup divergence and low intrahaplogroup polymorphism. Using maximum likelihood models we find that the branches leading to these three distinct mitochondrial groups show a significantly reduced rate of nonsynonymous relative to synonymous substitution. This interhaplogroup rate is significantly reduced compared to the intrahaplogroup rate, and closely resembles the rate observed between distinct species. The data suggest that slightly deleterious mutations segregating within D. simulans haplogroups are removed by selection prior to their fixation among haplogroups. We explore several hypotheses to explain how lineages within a single species can be compatible with this model of slightly deleterious mutation. The most likely hypothesis is that D. simulans haplogroups have persisted in isolation, perhaps due to association with the bacterial symbiont Wolbachia and/or demographic history, introducing a bias against the fixation of slightly deleterious mutations.}, } @article {pmid15944085, year = {2005}, author = {Xi, Z and Dean, JL and Khoo, C and Dobson, SL}, title = {Generation of a novel Wolbachia infection in Aedes albopictus (Asian tiger mosquito) via embryonic microinjection.}, journal = {Insect biochemistry and molecular biology}, volume = {35}, number = {8}, pages = {903-910}, pmid = {15944085}, issn = {0965-1748}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Bacterial Infections/veterinary ; Cytoplasm ; Embryo, Nonmammalian ; Female ; Insect Vectors ; Male ; Microinjections/methods ; Mosquito Control/*methods ; Wolbachia/*pathogenicity ; }, abstract = {Genetic strategies that reduce or block pathogen transmission by mosquitoes are being investigated as a means to augment current control measures. Strategies of vector suppression and replacement are based upon intracellular Wolbachia bacteria, which occur naturally in many insect populations. Maternally inherited Wolbachia have evolved diverse mechanisms to manipulate host insect reproduction and promote infection invasion. One mechanism is cytoplasmic incompatibility (CI) through which Wolbachia promotes infection spread by effectively sterilizing uninfected females. In a prior field test, releases of Wolbachia-infected males were used to suppress a field population of Culex pipiens. An additional strategy would employ Wolbachia as a vehicle to drive desired transgenes into vector populations (population replacement). Wolbachia-based population suppression and population replacement strategies require an ability to generate artificial Wolbachia associations in mosquitoes. Here, we demonstrate a technique for transferring Wolbachia (transfection) in a medically important mosquito species: Aedes albopictus (Asian tiger mosquito). Microinjection was used to transfer embryo cytoplasm from a double-infected Ae. albopictus line into an aposymbiotic line. The resulting mosquito line is single-infected with the wAlbB Wolbachia type. The artificially generated infection type is not known to occur naturally and displays a new CI crossing type and the first known example of bidirectional CI in Aedes mosquitoes. We discuss the results in relation to applied mosquito control strategies and the evolution of Wolbachia infections in Ae. albopictus.}, } @article {pmid15937134, year = {2005}, author = {Clark, ME and Anderson, CL and Cande, J and Karr, TL}, title = {Widespread prevalence of wolbachia in laboratory stocks and the implications for Drosophila research.}, journal = {Genetics}, volume = {170}, number = {4}, pages = {1667-1675}, pmid = {15937134}, issn = {0016-6731}, mesh = {Animals ; Crosses, Genetic ; Drosophila/*growth & development/*microbiology ; Female ; Life Cycle Stages ; Male ; Microscopy, Confocal ; Models, Biological ; *Prevalence ; Reproduction/physiology ; *Research ; Rickettsiaceae Infections/*epidemiology ; Spermatogenesis ; *Wolbachia ; }, abstract = {Wolbachia is an intracellular microbe harbored by a wide variety of arthropods (including Drosophila) and filarial nematodes. Employing several different strategies including male killing, induced parthenogenesis, cytoplasmic incompatibility, and feminization, and acting by as-yet-unknown mechanisms, Wolbachia alters host reproduction to increase its representation within a population. Wolbachia is closely associated with gametic incompatibility but also interacts with Drosophila in other, little understood ways. We report here significant and widespread infection of Wolbachia within laboratory stocks and its real and potential impact on Drosophila research. We describe the results of a survey indicating that approximately 30% of stocks currently housed at the Bloomington Drosophila Stock Center are infected with Wolbachia. Cells of both reproductive tissues and numerous somatic organs harbor Wolbachia and display considerable variation in infection levels within and between both tissue types. These results are discussed from the perspective of Wolbachia's potential confounding effects on both host fitness and phenotypic analyses. In addition to this cautionary message, the infection status of stock centers may provide further opportunities to study the genetic basis of host/symbiosis.}, } @article {pmid15936655, year = {2005}, author = {Ogata, H and Suhre, K and Claverie, JM}, title = {Discovery of protein-coding palindromic repeats in Wolbachia.}, journal = {Trends in microbiology}, volume = {13}, number = {6}, pages = {253-255}, doi = {10.1016/j.tim.2005.03.013}, pmid = {15936655}, issn = {0966-842X}, mesh = {Amino Acid Sequence ; Bacterial Proteins/chemistry/*genetics ; Molecular Sequence Data ; *Open Reading Frames ; *Repetitive Sequences, Nucleic Acid ; Sequence Alignment ; Wolbachia/*genetics ; }, } @article {pmid15935392, year = {2005}, author = {Schofield, P and Chaplain, M and Hubbard, S}, title = {Evolution of searching and life history characteristics in individual-based models of host-parasitoid-microbe associations.}, journal = {Journal of theoretical biology}, volume = {237}, number = {1}, pages = {1-16}, doi = {10.1016/j.jtbi.2005.03.025}, pmid = {15935392}, issn = {0022-5193}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Bacterial Physiological Phenomena ; *Biological Evolution ; *Ecosystem ; Host-Parasite Interactions ; Insecta/parasitology/*physiology ; Life Cycle Stages ; Models, Biological ; Movement ; Parasites/microbiology/*physiology ; Population Dynamics ; Reproduction ; Sex Distribution ; Wolbachia/physiology ; }, abstract = {In this paper we develop a novel discrete, individual-based mathematical model of the evolution of life history, dispersal and other behavioural characteristics in insect host-parasitoid-microbe associations, and use it to investigate their evolutionary dynamics. For any individual characteristic the model begins with an even, rectangular distribution of characteristic values. Selection is then allowed to act, and the change in the distribution of the characteristic values is observed. Evolutionary change in the population variance of the characteristic value is also observed, since we would expect this to decline under selection in most cases. The paper, therefore, introduces a general framework for modeling problems of evolution in stochastic, spatially structured environments, where movement and dispersal are under selection. The model then extends this approach to include the sex-distorting bacterium Wolbachia in order to investigate aspects of its horizontal and vertical transmission under different levels of superparasitism by parasitoids. The model also includes a neutral genetic marker, in order to be able to detect changes in phenotype frequency caused by genetic drift, as well as a simplified simulation of sexual reproduction so as to allow the possibility of recombination between genotypes. Key results from the model simulations show that: (i) the refractory time after oviposition affects the value of superparasitism, with short refractory times favouring high rates of superparasitism; (ii) variable levels of superparasitism do not affect the stable proportion of the population of parasitoids infected with Wolbachia, but this is achieved by different evolutionary pathways under low and high superparasitism, respectively. In the case of low superparasitism Wolbachia spreads mainly by vertical transmission, leading to population replacement, whereas when superparasitism rates are high there is significant horizontal transfer.}, } @article {pmid15933022, year = {2005}, author = {Xi, Z and Dobson, SL}, title = {Characterization of Wolbachia transfection efficiency by using microinjection of embryonic cytoplasm and embryo homogenate.}, journal = {Applied and environmental microbiology}, volume = {71}, number = {6}, pages = {3199-3204}, pmid = {15933022}, issn = {0099-2240}, support = {R01 AI051533/AI/NIAID NIH HHS/United States ; NIH-AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Buffers ; Cytoplasm/physiology ; Drosophila/*embryology/*microbiology ; *Microinjections ; Wolbachia/*physiology ; }, abstract = {Wolbachia spp. are intracellular alpha proteobacteria closely related to Rickettsia. The maternally inherited infections occur in a wide range of invertebrates, causing several reproductive abnormalities, including cytoplasmic incompatibility. The artificial transfer of Wolbachia between hosts (transfection) is used both for basic research examining the Wolbachia-host interaction and for applied strategies that use Wolbachia infections to affect harmful insect populations. Commonly employed transfection techniques use embryonic microinjection to transfer Wolbachia-infected embryo cytoplasm or embryo homogenate. Although microinjections of both embryonic cytoplasm and homogenate have been used successfully, their respective transfection efficiencies (rates of establishing stable germ line infections) have not been directly compared. Transfection efficiency may be affected by variation in Wolbachia quantity or quality within the donor embryos and/or the buffer types used in embryo homogenization. Here we have compared Wolbachia bacteria that originate from different embryonic regions for their competencies in establishing stable germ line infections. The following three buffers were compared for their abilities to maintain an appropriate in vitro environment for Wolbachia during homogenization and injection: phosphate-buffered saline, Drosophila Ringer's buffer, and a sucrose-phosphate-glutamate solution (SPG buffer). The results demonstrate that Wolbachia bacteria from both anterior and posterior embryo cytoplasms are competent for establishing infection, although differing survivorships of injected hosts were observed. Buffer comparison shows that embryos homogenized in SPG buffer yielded the highest transfection success. No difference was observed in transfection efficiencies when the posterior cytoplasm transfer and SPG-homogenized embryo techniques were compared. We discuss the results in relation to intra- and interspecific Wolbachia transfection and the future adaptation of the microinjection technique for additional insects.}, } @article {pmid15931253, year = {2005}, author = {Zeh, DW and Zeh, JA and Bonilla, MM}, title = {Wolbachia, sex ratio bias and apparent male killing in the harlequin beetle riding pseudoscorpion.}, journal = {Heredity}, volume = {95}, number = {1}, pages = {41-49}, doi = {10.1038/sj.hdy.6800666}, pmid = {15931253}, issn = {0018-067X}, support = {P20 RR16464/RR/NCRR NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/pharmacology ; Arachnida/*parasitology ; Female ; Inheritance Patterns ; Larva ; Male ; Mortality ; Selection, Genetic ; Tetracycline/administration & dosage/pharmacology ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {Bacterial endosymbionts that manipulate host reproduction are now known to be widespread in insects and other arthropods. Since they inhabit the cytoplasm and are maternally inherited, these microorganisms can enhance their fitness by biasing host sex ratio in favour of females. At its most extreme, sex ratio manipulation may be achieved by killing male embryos, as occurs in a number of insect species. Here, we provide evidence for the first case of male killing by a tetracycline-sensitive microbe in pseudoscorpions. Using a combination of inheritance studies, antibiotic treatment and molecular assays, we show that a new strain of Wolbachia is associated with extreme female bias in the pseudoscorpion, Cordylochernes scorpioides. In a highly female-biased line, sex ratio distortion was maternally inherited, and occurred in conjunction with a high rate of spontaneous abortion and low reproductive success. Antibiotic treatment cured females of the Wolbachia infection, restored offspring sex ratio to 1:1, and significantly enhanced female reproductive success. The discovery of apparent male-killing in C. scorpioides is of interest because pseudoscorpions are viviparous. Theory predicts that male killing should be favoured, if male death enhances the fitness of infected female siblings. In a live-bearing host, reallocation of maternal resources from dead male embryos to their sisters provides a direct, physiological mechanism through which fitness compensation could favour male killing by cellular endosymbionts. Our results suggest, however, that fitness compensation and the spread of male-killing endosymbionts may be undermined by a high rate of spontaneous abortion in infected females of this viviparous arthropod.}, } @article {pmid15931237, year = {2005}, author = {van der Giezen, M}, title = {Endosymbiosis: past and present.}, journal = {Heredity}, volume = {95}, number = {5}, pages = {335-336}, doi = {10.1038/sj.hdy.6800703}, pmid = {15931237}, issn = {0018-067X}, mesh = {Buchnera/genetics ; DNA, Mitochondrial/genetics ; *Evolution, Molecular ; Symbiosis/*genetics ; Wolbachia/genetics ; }, } @article {pmid15927071, year = {2005}, author = {Ahrens, ME and Shoemaker, D}, title = {Evolutionary history of Wolbachia infections in the fire ant Solenopsis invicta.}, journal = {BMC evolutionary biology}, volume = {5}, number = {}, pages = {35}, pmid = {15927071}, issn = {1471-2148}, mesh = {Animals ; Ants/*microbiology ; Bacterial Infections/*genetics ; DNA, Mitochondrial/*genetics/metabolism ; Evolution, Molecular ; *Genes, Bacterial ; Genetic Variation ; Genetics, Population ; Haplotypes ; Models, Biological ; Phylogeny ; Time Factors ; Wolbachia/*metabolism ; }, abstract = {BACKGROUND: Wolbachia are endosymbiotic bacteria that commonly infect numerous arthropods. Despite their broad taxonomic distribution, the transmission patterns of these bacteria within and among host species are not well understood. We sequenced a portion of the wsp gene from the Wolbachia genome infecting 138 individuals from eleven geographically distributed native populations of the fire ant Solenopsis invicta. We then compared these wsp sequence data to patterns of mitochondrial DNA (mtDNA) variation of both infected and uninfected host individuals to infer the transmission patterns of Wolbachia in S. invicta.

RESULTS: Three different Wolbachia (wsp) variants occur within S. invicta, all of which are identical to previously described strains in fire ants. A comparison of the distribution of Wolbachia variants within S. invicta to a phylogeny of mtDNA haplotypes suggests S. invicta has acquired Wolbachia infections on at least three independent occasions. One common Wolbachia variant in S. invicta (wSinvictaB) is associated with two divergent mtDNA haplotype clades. Further, within each of these clades, Wolbachia-infected and uninfected individuals possess virtually identical subsets of mtDNA haplotypes, including both putative derived and ancestral mtDNA haplotypes. The same pattern also holds for wSinvictaA, where at least one and as many as three invasions into S. invicta have occurred. These data suggest that the initial invasions of Wolbachia into host ant populations may be relatively ancient and have been followed by multiple secondary losses of Wolbachia in different infected lineages over time. Finally, our data also provide additional insights into the factors responsible for previously reported variation in Wolbachia prevalence among S. invicta populations.

CONCLUSION: The history of Wolbachia infections in S. invicta is rather complex and involves multiple invasions or horizontal transmission events of Wolbachia into this species. Although these Wolbachia infections apparently have been present for relatively long time periods, these data clearly indicate that Wolbachia infections frequently have been secondarily lost within different lineages. Importantly, the uncoupled transmission of the Wolbachia and mtDNA genomes suggests that the presumed effects of Wolbachia on mtDNA evolution within S. invicta are less severe than originally predicted. Thus, the common concern that use of mtDNA markers for studying the evolutionary history of insects is confounded by maternally inherited endosymbionts such as Wolbachia may be somewhat unwarranted in the case of S. invicta.}, } @article {pmid15926897, year = {2005}, author = {Montenegro, H and Solferini, VN and Klaczko, LB and Hurst, GD}, title = {Male-killing Spiroplasma naturally infecting Drosophila melanogaster.}, journal = {Insect molecular biology}, volume = {14}, number = {3}, pages = {281-287}, doi = {10.1111/j.1365-2583.2005.00558.x}, pmid = {15926897}, issn = {0962-1075}, mesh = {Animals ; Bacterial Proteins/genetics ; Base Sequence ; Drosophila melanogaster/embryology/*microbiology ; Male ; Phylogeny ; Sex Factors ; Sex Ratio ; Spiroplasma/classification/*physiology ; Wolbachia/genetics ; }, abstract = {Elucidation of the mechanism of action of selfish genetic elements is difficult outside species with well-defined genetics. Male-killing, the phenomenon whereby inherited bacteria kill male hosts during embryogenesis, is thus uncharacterized in mechanistic terms despite being common and important in insects. We characterized the prevalence, identity and source of the male-killing infection recently discovered in Drosophila melanogaster in Brazil. Male-killing was found to be present in 2.3% of flies from Recife, Brazil, and was uniquely associated with the presence of Spiroplasma infection. The identity of sequences across part of the 16S and across the 16S-23S ITS region indicated that the male-killing infection of D. melanogaster was very closely related to S. poulsonii, the source of the male-killing infection in willistoni group flies also found in South America. The sequences of two further protein-coding genes indicated the D. melanogaster infection to be most closely related to that found in D. nebulosa, from the willistoni group. Our data suggest that the establishment of D. melanogaster in South America was associated with the movement of male-killing bacteria between species.}, } @article {pmid15926693, year = {2005}, author = {Dyer, KA and Minhas, MS and Jaenike, J}, title = {Expression and modulation of embryonic male-killing in Drosophila innubila: opportunities for multilevel selection.}, journal = {Evolution; international journal of organic evolution}, volume = {59}, number = {4}, pages = {838-848}, pmid = {15926693}, issn = {0014-3820}, mesh = {Animals ; Arizona ; DNA Primers ; Drosophila/microbiology/*physiology ; Female ; Male ; Mortality ; Polymerase Chain Reaction/methods ; Population Density ; *Selection, Genetic ; *Sex Ratio ; *Symbiosis ; Temperature ; Tetracycline/toxicity ; Wolbachia/drug effects/genetics/*physiology ; }, abstract = {Organisms and the symbionts they harbor may experience opposing forces of selection. In particular, the contrasting inheritance patterns of maternally transmitted symbionts and their host's nuclear genes can engender conflict among organizational levels over the optimal host offspring sex ratio. This study uses a male-killing Wolbachia endosymbiont and its host Drosophila innubila to experimentally address the potential for multilevel selection in a host-symbiont system. We show that bacterial density can vary among infected females, and that females with a higher density have a more female-biased offspring sex ratio. Furthermore, bacterial density is an epigenetic and heritable trait: females with a low bacterial load have daughters with a lower-than-average bacterial density, whose offspring then experience less severe male-killing. For infected sons, the probability of embryonic mortality increases with the bacterial density in their mothers. The frequency distribution of Wolbachia density among individual D. innubila females, and therefore the dynamics of infection within populations of these flies, results both from processes affecting the growth and regulation of bacterial populations within cytoplasmic lineages and from selection among cytoplasmic lineages that vary in bacterial density. Estimates of effective population size of Wolbachia within cytoplasmic lineages and of D. innubila at the host population level suggest that selection among cytoplasmic lineages is likely to overwhelm the results of selection within lineages.}, } @article {pmid15894186, year = {2005}, author = {Aksoy, S and Rio, RV}, title = {Interactions among multiple genomes: tsetse, its symbionts and trypanosomes.}, journal = {Insect biochemistry and molecular biology}, volume = {35}, number = {7}, pages = {691-698}, doi = {10.1016/j.ibmb.2005.02.012}, pmid = {15894186}, issn = {0965-1748}, support = {AI051584/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Host-Parasite Interactions ; Humans ; Insect Vectors/*genetics ; Symbiosis ; Trypanosoma/*genetics ; Trypanosomiasis/immunology/prevention & control ; Tsetse Flies/*genetics/microbiology ; }, abstract = {Insect-borne diseases exact a high public health burden and have a devastating impact on livestock and agriculture. To date, control has proved to be exceedingly difficult. One such disease that has plagued sub-Saharan Africa is caused by the protozoan African trypanosomes (Trypanosoma species) and transmitted by tsetse flies (Diptera: Glossinidae). This presentation describes the biology of the tsetse fly and its interactions with trypanosomes as well as its symbionts. Tsetse can harbor up to three distinct microbial symbionts, including two enterics (Wigglesworthia glossinidia and Sodalis glossinidius) as well as facultative Wolbachia infections, which influence host physiology. Recent investigations into the genome of the obligate symbiont Wigglesworthia have revealed characteristics indicative of its long co-evolutionary history with the tsetse host species. Comparative analysis of the commensal-like Sodalis with free-living enterics provides examples of adaptations to the host environment (physiology and ecology), reflecting genomic tailoring events during the process of transitioning into a symbiotic lifestyle. From an applied perspective, the extensive knowledge accumulated on the genomic and developmental biology of the symbionts coupled with our ability to both express foreign genes in these microbes in vitro and repopulate tsetse midguts with these engineered microbes now provides a means to interfere with the host physiological traits which contribute to vector competence promising a novel tool for disease management.}, } @article {pmid15885912, year = {2005}, author = {Kramer, L and Simón, F and Tamarozzi, F and Genchi, M and Bazzocchi, C}, title = {Is Wolbachia complicating the pathological effects of Dirofilaria immitis infections?.}, journal = {Veterinary parasitology}, volume = {133}, number = {2-3}, pages = {133-136}, doi = {10.1016/j.vetpar.2005.04.011}, pmid = {15885912}, issn = {0304-4017}, mesh = {Animals ; Antibodies, Bacterial/analysis/biosynthesis ; Bacterial Outer Membrane Proteins/*immunology/isolation & purification ; Dirofilaria immitis/*microbiology ; Dirofilariasis/*immunology/*microbiology/parasitology ; Dog Diseases/immunology/microbiology/parasitology ; Dogs ; Immunoglobulin G/analysis/biosynthesis ; Immunohistochemistry/veterinary ; Wolbachia/*immunology ; }, abstract = {Human and animal parasitic filarial nematodes, which often are the cause of severe disease, harbor intracellular bacteria of the genus Wolbachia (Rickettsiaceae). It is thought that these bacteria play an important role in the pathogenesis and immune response to filarial infection. In order to determine the possible role of Wolbachia in heartworm disease, dogs naturally infected with Dirofilaria immitis were studied for specific antibody response to Wolbachia surface protein (WSP). Antibody subclasses were analyzed to determine immune response polarization. Dogs that died from heartworm disease were necropsied, and various organs were studied by immunohistochemistry to determine whether Wolbachia-derived molecules were present in tissue from infected dogs. Humoral response to the WSP was present in all infected dogs and appeared to be predominantly of the Th1-type. Several organs, including lung, liver, and kidney, contained positive-staining cells for WSP, confirming that the canine host does come into contact with Wolbachia-derived molecules.}, } @article {pmid15876457, year = {2005}, author = {Kramer, LH and Tamarozzi, F and Morchón, R and López-Belmonte, J and Marcos-Atxutegi, C and Martín-Pacho, R and Simón, F}, title = {Immune response to and tissue localization of the Wolbachia surface protein (WSP) in dogs with natural heartworm (Dirofilaria immitis) infection.}, journal = {Veterinary immunology and immunopathology}, volume = {106}, number = {3-4}, pages = {303-308}, doi = {10.1016/j.vetimm.2005.03.011}, pmid = {15876457}, issn = {0165-2427}, mesh = {Animals ; Antibodies, Bacterial/biosynthesis ; Bacterial Outer Membrane Proteins/*immunology/metabolism ; Dirofilaria immitis/*microbiology ; Dirofilariasis/*immunology/*microbiology/parasitology ; Dogs ; Female ; Humans ; Immunoglobulin G/biosynthesis ; Immunohistochemistry ; Wolbachia/*immunology ; }, abstract = {Human and animal parasitic filarial nematodes, including the agent of canine and feline heartworm disease Dirofilaria immitis, harbour intracellular bacteria of the genus Wolbachia (Rickettsiaies). It is thought that these bacteria play an important role in the pathogenesis and immune response to filarial infection. Immunoglobulin G (total IgG, IgG1, IgG2) production against and immunohistochemical staining of tissues for the Wolbachia surface protein (WSP) from dogs with natural heartworm infection were evaluated. All infected dogs had significant total anti-WSP IgG levels compared to healthy controls. Interestingly, WSP was recognized by the IgG2 subclass in both microfilariemic dogs and in dogs with no circulating microfilariae (occult infection). However, microfilariemic dogs also produced gG1 antibodies. Positive staining for WSP was observed in lungs, liver and kidneys, in particular in glomerular capillaries of naturally infected dogs who had died from heartworm disease. Our results show for the first time that Wolbachia is recognized specifically by D. immitis--infected dogs and that the bacteria is released into host tissue. Furthermore, microfilariemic status appears to effect immune responses to this endosymbiont.}, } @article {pmid15865286, year = {2005}, author = {Shaikevich, EV and Vinogradova, EB and Platonov, AE and Karan, LS and Zakharov, IA}, title = {[Polymorphism of mitochondrial DNA and infection with symbiotic cytoplasmic bacterium Wolbachia pipientis in mosquitoes of the Culex pipiens complex from Russia].}, journal = {Genetika}, volume = {41}, number = {3}, pages = {320-325}, pmid = {15865286}, issn = {0016-6758}, mesh = {Animals ; Culex/*genetics/*microbiology ; DNA, Mitochondrial/*genetics ; *Polymorphism, Genetic ; Russia ; Species Specificity ; Symbiosis/*genetics/physiology ; Wolbachia/*physiology ; }, abstract = {A total of 208 mosquitoes of the Culex pipiens complex from 15 basement and terrestrial populations collected in different regions of the European part of Russia and Siberia were examined by genetic methods. Among these, two major mitotypes, M and P, were identified. These mitotypes differed by six substitutions in the 246-bp mitochondrial DNA cytochrome oxidase I gene fragment examined. Urban basement mosquito ecotype C. pipiens (form molestus) were characterized by the presence of mitotype M and infection with the endosymbiotic bacteria of the genus Wolbachia. Mosquitoes of the C. pipiens complex inhabiting opened biotopes harbored mitotype P, or its variety, mitotype P1, and were not infected with Wolbachia. Thus, in natural conditions marked linkage disequilibrium between cytoplasmic elements, mitochondrial DNA and Wolbachia, can be observed. Similarity of mitotypes in form molestus mosquito from different geographical localities favors the hypothesis on the common ancestry of urban mosquitoes.}, } @article {pmid15862594, year = {2005}, author = {Telschow, A and Yamamura, N and Werren, JH}, title = {Bidirectional cytoplasmic incompatibility and the stable coexistence of two Wolbachia strains in parapatric host populations.}, journal = {Journal of theoretical biology}, volume = {235}, number = {2}, pages = {265-274}, doi = {10.1016/j.jtbi.2005.01.008}, pmid = {15862594}, issn = {0022-5193}, mesh = {Animals ; Arthropods/genetics/*microbiology ; *Biological Evolution ; Cytoplasm/*microbiology ; Female ; Models, Biological ; *Models, Genetic ; Rickettsiaceae Infections/transmission/veterinary ; Species Specificity ; Symbiosis ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachiaare intracellular bacteria which are very widely distributed among arthropods. In many insect species Wolbachiaare known to induce cytoplasmic mating incompatibility (CI). It has been suggested that Wolbachiacould promote speciation in their hosts if parapatric host populations are infected with two different Wolbachiastrains causing bidirectional mating incompatibilities. A necessary condition for this speciation scenario to work is that the two Wolbachiastrains can stably coexist. The following study investigates this problem analysing a mathematical model with two host populations and migration between them. We show that the stability of bidirectional CI can be fully described in terms of a critical migration rate which is defined as the highest migration below which a stable coexistence of two Wolbachiastrains is possible. For some special cases we could derive analytical solutions for the critical migration rate; for the general case estimations of the critical migration rate are given. Our main finding is that bidirectional CI can stably persist in the face of high migration and can be as high as over 15% per generation for CI levels observed in nature. These results have implications for the potential of Wolbachiato promote genetic divergence and speciation in their hosts.}, } @article {pmid15839745, year = {2005}, author = {Pfarr, K and Hoerauf, A}, title = {The annotated genome of Wolbachia from the filarial nematode Brugia malayi: what it means for progress in antifilarial medicine.}, journal = {PLoS medicine}, volume = {2}, number = {4}, pages = {e110}, pmid = {15839745}, issn = {1549-1676}, mesh = {Animals ; Brugia malayi/*microbiology/*pathogenicity ; Filariasis/*drug therapy/prevention & control ; Filaricides/*pharmacology ; Genome ; Humans ; Phenotype ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {Filarial nematodes contain endosymbiotic bacteria of the genus Wolbachia. As described in the April 2005 issue of PLoS Biology, Foster et al. have sequenced the genome of the Wolbachia that lives in the nematode Brugia malayi. What are the clinical implications?}, } @article {pmid15834154, year = {2005}, author = {Charlat, S and Calmet, C and Andrieu, O and Merçot, H}, title = {Exploring the evolution of Wolbachia compatibility types: a simulation approach.}, journal = {Genetics}, volume = {170}, number = {2}, pages = {495-507}, pmid = {15834154}, issn = {0016-6731}, mesh = {Animals ; Biological Evolution ; Chromosomes ; Computer Simulation ; Crosses, Genetic ; Cytoplasm/metabolism ; Evolution, Molecular ; Female ; Genetic Variation ; Host-Parasite Interactions ; Male ; Models, Genetic ; Models, Theoretical ; Mutation ; Polymorphism, Genetic ; Recombination, Genetic ; Reproduction/physiology ; Wolbachia/*genetics ; }, abstract = {Wolbachia-induced cytoplasmic incompatibility (CI) is observed when males bearing the bacterium mate with uninfected females or with females bearing a different Wolbachia variant; in such crosses, paternal chromosomes are lost at the first embryonic mitosis, most often resulting in developmental arrest. The molecular basis of CI is currently unknown, but it is useful to distinguish conceptually the male and female sides of this phenomenon: in males, Wolbachia must do something, before it is shed from maturing sperm, that will disrupt paternal chromosomes functionality [this is usually termed "the modification (mod) function"]; in females, Wolbachia must somehow restore embryonic viability, through what is usually called "the rescue (resc) function." The occurrence of CI in crosses between males and females bearing different Wolbachia variants demonstrates that the mod and resc functions interact in a specific manner: different mod resc pairs make different compatibility types. We are interested in the evolutionary process allowing the diversification of compatibility types. In an earlier model, based on the main assumption that the mod and resc functions can mutate independently, we have shown that compatibility types can evolve through a two-step process, the first involving drift on mod variations and the second involving selection on resc variations. This previous study has highlighted the need for simulation-based models that would include the effects of nondeterministic evolutionary forces. This study is based on a simulation program fulfilling this condition, allowing us to follow the evolution of compatibility types under mutation, drift, and selection. Most importantly, simulations suggest that in the frame of our model, the evolution of compatibility types is likely to be a gradual process, with new compatibility types remaining partially compatible with ancestral ones.}, } @article {pmid15828434, year = {2004}, author = {Bazzocchi, C and Lecchi, C and Kramer, LH and Genchi, C and Bandi, C}, title = {Sequencing of the complete gene coding for the GroEL of the Wolbachia of Dirofilaria immitis and expression and purification of the recombinant protein.}, journal = {Parassitologia}, volume = {46}, number = {3}, pages = {307-310}, pmid = {15828434}, issn = {0048-2951}, mesh = {Animals ; Bacterial Proteins/*genetics/isolation & purification ; Chaperonin 60/*genetics/isolation & purification ; Chromatography, High Pressure Liquid ; DNA, Bacterial/genetics ; Dirofilaria immitis/*microbiology ; Female ; Genes, Bacterial ; Molecular Sequence Data ; Polymerase Chain Reaction ; Recombinant Fusion Proteins/isolation & purification ; Sequence Analysis, DNA ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Wolbachia are intracellular bacteria that infect arthropods and filarial nematodes. These bacteria play an important role in the immunology and pathogenesis of filarial diseases through their proteins and, possibly, other molecules. GroEL is a constitutively expressed bacterial protein; it is highly conserved among bacteria and is involved in the correct folding of newly synthesized proteins. Here we report the production of recombinant GroEL from the Wolbachia of Dirofilaria immitis. Our goal is to test the hypothesis that GroEL is involved in the immunopathology of filariases. The complete groel gene was PCR-amplified, sequenced and cloned into an expression vector. The recombinant GroEL was purified by affinity chromatography by using high-performance liquid chromatography (HPLC).}, } @article {pmid15813794, year = {2005}, author = {Duron, O and Lagnel, J and Raymond, M and Bourtzis, K and Fort, P and Weill, M}, title = {Transposable element polymorphism of Wolbachia in the mosquito Culex pipiens: evidence of genetic diversity, superinfection and recombination.}, journal = {Molecular ecology}, volume = {14}, number = {5}, pages = {1561-1573}, doi = {10.1111/j.1365-294X.2005.02495.x}, pmid = {15813794}, issn = {0962-1083}, mesh = {Amino Acid Sequence ; Animals ; Base Sequence ; Blotting, Southern ; Culex/*microbiology ; DNA Primers ; DNA Transposable Elements/*genetics ; *Demography ; Molecular Sequence Data ; *Polymorphism, Genetic ; Recombination, Genetic/*genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Species Specificity ; Wolbachia/*genetics ; }, abstract = {Wolbachia is a group of maternally inherited endosymbiotic bacteria that infect and induce cytoplasmic incompatibility (CI) in a wide range of arthropods. In contrast to other species, the mosquito Culex pipiens displays an extremely high number of CI types suggesting differential infection by multiple Wolbachia strains. Attempts so far failed to detect Wolbachia polymorphism that might explain this high level of CI diversity found in C. pipiens populations. Here, we establish that Wolbachia infection is near to or at fixation in worldwide populations of the C. pipiens complex. Wolbachia polymorphism was addressed by sequence analysis of the Tr1 gene, a unique transposable element of the IS5 family, which allowed the identification of five C. pipiens Wolbachia strains, differing either by nucleotide substitution, presence or absence pattern, or insertion site. Sequence analysis also showed that recombination, transposition and superinfection occurred at very low frequencies. Analysis of the geographical distributions of each Wolbachia strain among C. pipiens populations indicated a strong worldwide differentiation independent from mosquito subspecies type, except in the UK. The availability of this polymorphic marker now opens the way to investigate evolution of Wolbachia populations and CI dynamics, in particular in regions where multiple crossing types coexist among C. pipiens populations.}, } @article {pmid15813792, year = {2005}, author = {Dedeine, F and Ahrens, M and Calcaterra, L and Shoemaker, DD}, title = {Social parasitism in fire ants (Solenopsis spp.): a potential mechanism for interspecies transfer of Wolbachia.}, journal = {Molecular ecology}, volume = {14}, number = {5}, pages = {1543-1548}, doi = {10.1111/j.1365-294X.2005.02499.x}, pmid = {15813792}, issn = {0962-1083}, mesh = {Animals ; Ants/microbiology/*physiology ; Bacterial Outer Membrane Proteins/*genetics ; Base Sequence ; Bayes Theorem ; Cluster Analysis ; *Genetic Variation ; Models, Genetic ; Molecular Sequence Data ; *Phylogeny ; Sequence Analysis, DNA ; *Social Behavior ; Species Specificity ; Wolbachia/*genetics ; }, abstract = {One possible mechanism for interspecific transfer of Wolbachia is through the intimate contact between parasites and their hosts. We surveyed 10 species of fly parasitoids (Pseudacteon spp.) and one inquiline social parasite, Solenopsis daguerrei, for the presence and sequence identity (wsp gene) of Wolbachia. Two Wolbachia variants infecting S. daguerrei were identical to known variants infecting the two common ant host species, Solenopsis invicta and Solenopsis richteri, suggesting possible transfers of Wolbachia between this parasite and their hosts have occurred. Our data also revealed an unexpectedly high diversity of Wolbachia variants within S. daguerrei: up to eight variants were found within each individual, which, to our knowledge, is the highest reported number of Wolbachia variants infecting a single individual of any host species.}, } @article {pmid15811532, year = {2005}, author = {Higazi, TB and Filiano, A and Katholi, CR and Dadzie, Y and Remme, JH and Unnasch, TR}, title = {Wolbachia endosymbiont levels in severe and mild strains of Onchocerca volvulus.}, journal = {Molecular and biochemical parasitology}, volume = {141}, number = {1}, pages = {109-112}, doi = {10.1016/j.molbiopara.2005.02.006}, pmid = {15811532}, issn = {0166-6851}, mesh = {Animals ; DNA, Bacterial/analysis ; Female ; Humans ; Male ; Onchocerca volvulus/isolation & purification/microbiology/*pathogenicity ; Onchocerciasis, Ocular/etiology ; Symbiosis ; Virulence ; Wolbachia/genetics/*physiology ; }, abstract = {Epidemiological, clinical and genetic data have all suggested that the filarial parasite Onchocerca volvulus, the causative agent of onchocerciasis (or river blindness) exists as two strains in West Africa. The severe strain induces severe ocular disease in a large proportion of the infected population, while the mild strain induces little ocular disease. Although DNA probes based upon a non-coding repeat sequence family can distinguish the two strains, the underlying basis for this difference in pathogenicity is not understood. Recently, several studies have implicated products produced by the Wolbachia endosymbiotic bacterium of O. volvulus in the pathogenesis of onchocerciasis. This suggested the hypothesis that differences in the Wolbachia endosymbiont population might be responsible for the pathogenic differences noted in the two strains. To test this hypothesis, quantitative PCR assays were used to measure the amount of Wolbachia DNA per nuclear genome in a collection of well characterized samples of mild and severe strain O. volvulus. The median ratio of Wolbachia DNA to nuclear DNA was significantly greater in severe strain parasites than in mild strain parasites. These data support the hypothesis that the pathogenic differences seen in severe and mild strain O. volvulus may be a function of their relative Wolbachia burden and provide additional support to the hypothesis that Wolbachia products may play a central role in the pathogenesis of ocular onchocerciasis.}, } @article {pmid15807436, year = {2005}, author = {Dudkina, NV and Kiseleva, EV}, title = {[Structural organization and distribution of the symbiotic bacteria Wolbachia during spermatogenesis of Drosophila simulans].}, journal = {Ontogenez}, volume = {36}, number = {1}, pages = {41-50}, pmid = {15807436}, issn = {0475-1450}, mesh = {Animals ; Drosophila/microbiology/physiology/*ultrastructure ; Male ; Meiosis ; Microscopy, Electron, Transmission ; Microtubules/ultrastructure ; Mitochondria/ultrastructure ; Spermatogenesis ; Spermatozoa/microbiology/*ultrastructure ; Symbiosis ; Wolbachia/physiology/*ultrastructure ; }, abstract = {Electron microscopy and morphometric analysis have shown that the symbiotic bacteria Wolbachia occur the testis cells during spermatogenesis and are absent in mature spermatids. Bacteria did not affect the structural organization of testis cells, which have a typical morphology during morphogenesis. Bacteria were distributed along the meiotic spindle microtubules near the mitochondria. They increased in number in spermatids at the stage of elongation. Endosymbionts aggregated at the spermatid distal end and contained many vacuoles but were absent at the spermatid proximal end near the nuclei. It was shown for the first time that the diameter of spermatids in a strongly infected line was two of three times that in a noninfected line. We hypothesize that the increase in the number of endosymbionts during spermatid elongation can affect the chromatin condensation in the spermatozoon.}, } @article {pmid15799519, year = {2005}, author = {Chen, WJ and Tsai, KH and Cheng, SL and Huang, CG and Wu, WJ}, title = {Using in situ hybridization to detect endosymbiont Wolbachia in dissected tissues of mosquito host.}, journal = {Journal of medical entomology}, volume = {42}, number = {2}, pages = {120-124}, doi = {10.1093/jmedent/42.2.120}, pmid = {15799519}, issn = {0022-2585}, mesh = {Aedes/*microbiology ; Animals ; Bacterial Outer Membrane Proteins/genetics ; DNA Probes ; *In Situ Hybridization ; Microscopy, Electron ; Salivary Glands/microbiology ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {The endosymbiont Wolbachia, extensively occurring in arthropods, usually causes reproductive distortions of the host, such as mosquitoes. In past years, detection of Wolbachia in host tissues has highly relied on transmission electron microscopy (TEM) that is tedious and usually unable to gain satisfactory results without experienced techniques and expensive instruments. Polymerase chain reaction (PCR) recently has become popular in Wolbachia identification. However, necessity of DNA extraction from host individuals or dissected tissues has limited its application in extensiveness and versatility. At present, in situ hybridization has increased its role in examination of various microbes. This report provides a technique for rapid detection and localization of Wolbachia in tissues dissected from mosquitoes and possibly other infected organisms. To detect Wolbachia and to localize them in host tissues more precisely, in situ hybridization by using digoxigenin (DIG)-labeled probes was invented and applied to Wolbachia detection in this study. The results showed that Wolbachia preferentially aggregate in ovarioles, which is consistent with previous observations by TEM. The endobacteria also were detected in salivary glands, mostly in lateral lobes. Ultrastructurally, Wolbachia has been shown to occur in the cytoplasma of salivary gland cells.}, } @article {pmid15780005, year = {2005}, author = {Foster, J and Ganatra, M and Kamal, I and Ware, J and Makarova, K and Ivanova, N and Bhattacharyya, A and Kapatral, V and Kumar, S and Posfai, J and Vincze, T and Ingram, J and Moran, L and Lapidus, A and Omelchenko, M and Kyrpides, N and Ghedin, E and Wang, S and Goltsman, E and Joukov, V and Ostrovskaya, O and Tsukerman, K and Mazur, M and Comb, D and Koonin, E and Slatko, B}, title = {The Wolbachia genome of Brugia malayi: endosymbiont evolution within a human pathogenic nematode.}, journal = {PLoS biology}, volume = {3}, number = {4}, pages = {e121}, pmid = {15780005}, issn = {1545-7885}, mesh = {Animals ; Brugia malayi/*genetics/pathogenicity ; *Evolution, Molecular ; Gene Expression Regulation, Bacterial ; *Genome, Bacterial ; Humans ; Molecular Sequence Data ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Complete genome DNA sequence and analysis is presented for Wolbachia, the obligate alpha-proteobacterial endosymbiont required for fertility and survival of the human filarial parasitic nematode Brugia malayi. Although, quantitatively, the genome is even more degraded than those of closely related Rickettsia species, Wolbachia has retained more intact metabolic pathways. The ability to provide riboflavin, flavin adenine dinucleotide, heme, and nucleotides is likely to be Wolbachia's principal contribution to the mutualistic relationship, whereas the host nematode likely supplies amino acids required for Wolbachia growth. Genome comparison of the Wolbachia endosymbiont of B. malayi (wBm) with the Wolbachia endosymbiont of Drosophila melanogaster (wMel) shows that they share similar metabolic trends, although their genomes show a high degree of genome shuffling. In contrast to wMel, wBm contains no prophage and has a reduced level of repeated DNA. Both Wolbachia have lost a considerable number of membrane biogenesis genes that apparently make them unable to synthesize lipid A, the usual component of proteobacterial membranes. However, differences in their peptidoglycan structures may reflect the mutualistic lifestyle of wBm in contrast to the parasitic lifestyle of wMel. The smaller genome size of wBm, relative to wMel, may reflect the loss of genes required for infecting host cells and avoiding host defense systems. Analysis of this first sequenced endosymbiont genome from a filarial nematode provides insight into endosymbiont evolution and additionally provides new potential targets for elimination of cutaneous and lymphatic human filarial disease.}, } @article {pmid15774024, year = {2005}, author = {Salzberg, SL and Dunning Hotopp, JC and Delcher, AL and Pop, M and Smith, DR and Eisen, MB and Nelson, WC}, title = {Serendipitous discovery of Wolbachia genomes in multiple Drosophila species.}, journal = {Genome biology}, volume = {6}, number = {3}, pages = {R23}, pmid = {15774024}, issn = {1474-760X}, support = {R01 LM006845/LM/NLM NIH HHS/United States ; R01 LM007938/LM/NLM NIH HHS/United States ; R01-LM007938/LM/NLM NIH HHS/United States ; R01-LM06845/LM/NLM NIH HHS/United States ; }, mesh = {Animals ; Base Sequence ; Drosophila/*microbiology ; Gene Transfer, Horizontal ; *Genome, Bacterial ; Genomics ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Wolbachia/classification/*genetics ; }, abstract = {BACKGROUND: The Trace Archive is a repository for the raw, unanalyzed data generated by large-scale genome sequencing projects. The existence of this data offers scientists the possibility of discovering additional genomic sequences beyond those originally sequenced. In particular, if the source DNA for a sequencing project came from a species that was colonized by another organism, then the project may yield substantial amounts of genomic DNA, including near-complete genomes, from the symbiotic or parasitic organism.

RESULTS: By searching the publicly available repository of DNA sequencing trace data, we discovered three new species of the bacterial endosymbiont Wolbachia pipientis in three different species of fruit fly: Drosophila ananassae, D. simulans, and D. mojavensis. We extracted all sequences with partial matches to a previously sequenced Wolbachia strain and assembled those sequences using customized software. For one of the three new species, the data recovered were sufficient to produce an assembly that covers more than 95% of the genome; for a second species the data produce the equivalent of a 'light shotgun' sampling of the genome, covering an estimated 75-80% of the genome; and for the third species the data cover approximately 6-7% of the genome.

CONCLUSIONS: The results of this study reveal an unexpected benefit of depositing raw data in a central genome sequence repository: new species can be discovered within this data. The differences between these three new Wolbachia genomes and the previously sequenced strain revealed numerous rearrangements and insertions within each lineage and hundreds of novel genes. The three new genomes, with annotation, have been deposited in GenBank.}, } @article {pmid15752546, year = {2005}, author = {Herbinière, J and Braquart-Varnier, C and Grève, P and Strub, JM and Frère, J and Van Dorsselaer, A and Martin, G}, title = {Armadillidin: a novel glycine-rich antibacterial peptide directed against gram-positive bacteria in the woodlouse Armadillidium vulgare (Terrestrial Isopod, Crustacean).}, journal = {Developmental and comparative immunology}, volume = {29}, number = {6}, pages = {489-499}, doi = {10.1016/j.dci.2004.11.001}, pmid = {15752546}, issn = {0145-305X}, mesh = {Amino Acid Motifs ; Amino Acid Sequence ; Animals ; Anti-Bacterial Agents/chemistry/*isolation & purification/pharmacology ; Base Sequence ; Blotting, Northern ; Gram-Positive Bacteria/drug effects/growth & development ; Gram-Positive Bacterial Infections/metabolism ; Hemocytes/metabolism ; Isopoda/chemistry/genetics/*metabolism/microbiology ; Microbial Sensitivity Tests ; Molecular Sequence Data ; *Peptides ; RNA/chemistry/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Analysis, Protein ; Spectrometry, Mass, Electrospray Ionization ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; }, abstract = {We report the isolation and the characterization of a novel antibacterial peptide from hemocytes of the woodlouse Armadillidium vulgare, naturally infected or uninfected by Wolbachia, an intracellular Gram-negative bacterium. This molecule displays antibacterial activity against Gram-positive bacteria despite its composition which classes it into the glycine-rich antibacterial peptide family, usually directed against fungi and Gram-negative bacteria. The complete sequence was determined by a combination of Edman degradation, mass spectrometry and cDNA cloning using a hemocyte library. The mature peptide (53 residues) has a 5259 Da molecular mass and is post-translationally modified by a C-terminal amidation. This peptide is characterized by a high level of glycine (47%) and a fivefold repeated motif GGGFH(R/S). As no evident sequence homology to other hitherto described antibacterial peptides has been found out, this antibacterial peptide was named armadillidin. Armadillidin is constitutively expressed in hemocytes and appears to be specific of A. vulgare.}, } @article {pmid15752179, year = {2005}, author = {Behbahani, A and Dutton, TJ and Davies, N and Townson, H and Sinkins, SP}, title = {Population differentiation and Wolbachia phylogeny in mosquitoes of the Aedes scutellaris group.}, journal = {Medical and veterinary entomology}, volume = {19}, number = {1}, pages = {66-71}, doi = {10.1111/j.0269-283X.2005.00542.x}, pmid = {15752179}, issn = {0269-283X}, mesh = {Aedes/genetics/*microbiology ; Animals ; Genetic Variation ; Genotype ; Phylogeny ; Wolbachia/*genetics/physiology ; }, abstract = {Mosquito species of the Aedes (Stegomyia) scutellaris (Walker) group (Diptera: Culicidae) are distributed across many islands of the South Pacific and include major regional vectors of filariasis, such as Aedes polynesiensis (Marks). Analysis of populations of Ae. polynesiensis at the extremes of its range, from Fiji and from Moorea, French Polynesia, using the rDNA ITS2 (internal transcribed spacer 2) region and six microsatellite markers showed considerable genetic differentiation between them (F(ST) = 0.298-0.357). Phylogenetic analysis of the Wolbachia endosymbionts in three members of the complex revealed that based on the wsp gene they are all very similar and belong to the Mel subgroup of the A clade, closely related to the Wolbachia strain present in the gall wasp Callyrhytis glandium (Giraud) (Hymenoptera: Cynipidae). By contrast they are only distantly related to the A-clade Wolbachia in Aedes albopictus (Skuse), a species closely allied to the Ae. scutellaris group. There was very low differentiation between the Wolbachia in the Moorea and Fiji populations of Ae. polynesiensis.}, } @article {pmid15752178, year = {2005}, author = {Dutton, TJ and Sinkins, SP}, title = {Filarial susceptibility and effects of Wolbachia in Aedes pseudoscutellaris mosquitoes.}, journal = {Medical and veterinary entomology}, volume = {19}, number = {1}, pages = {60-65}, doi = {10.1111/j.0269-283X.2005.00557.x}, pmid = {15752178}, issn = {0269-283X}, mesh = {Aedes/genetics/*microbiology/*parasitology ; Animals ; Anti-Bacterial Agents/pharmacology ; Brugia pahangi/*physiology ; Crosses, Genetic ; Female ; Host-Parasite Interactions ; Insect Vectors ; Larva/physiology ; Male ; Reproduction ; Tetracycline/pharmacology ; Wolbachia/drug effects/*physiology ; }, abstract = {The mosquito Aedes pseudoscutellaris (Theobald), a member of the Aedes (Stegomyia) scutellaris complex (Diptera: Culicidae), is an important vector of subperiodic Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae), causing human lymphatic filariasis, on South Pacific islands. Maternal inheritance of filarial susceptibility in the complex has previously been asserted, and larval tetracycline treatment reduced susceptibility; the maternally inherited Wolbachia in these mosquitoes were suggested to be responsible. To investigate the relationship of these two factors, we eliminated Wolbachia from a strain of Ae. pseudoscutellaris by tetracycline treatment, and tested filarial susceptibility of the adult female mosquitoes using Brugia pahangi (Edeson & Buckley). Filarial susceptibility was not significantly different in Wolbachia-free and infected lines of Ae. pseudoscutellaris, suggesting that the Wolbachia in these mosquitoes do not influence vector competence. Crosses between Wolbachia-infected males and uninfected females of Ae. pseudoscutellaris showed cytoplasmic incompatibility (CI), i.e. no eggs hatched, unaffected by larval crowding or restricted nutrient availability, whereas these factors are known to affect CI in Drosophila simulans. Reciprocal crosses between Ae. pseudoscutellaris and Ae. katherinensis Woodhill produced no progeny, even when both parents were Wolbachia-free, suggesting that nuclear factors are responsible for this interspecific sterility.}, } @article {pmid15750072, year = {2005}, author = {Sarih, M and M'Ghirbi, Y and Bouattour, A and Gern, L and Baranton, G and Postic, D}, title = {Detection and identification of Ehrlichia spp. in ticks collected in Tunisia and Morocco.}, journal = {Journal of clinical microbiology}, volume = {43}, number = {3}, pages = {1127-1132}, pmid = {15750072}, issn = {0095-1137}, mesh = {Anaplasmataceae/classification/*isolation & purification ; Animals ; Ehrlichia/classification/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Ticks/*microbiology ; }, abstract = {A broad-range 16S rRNA gene PCR assay followed by partial sequencing of the 16S rRNA gene was used for the detection of members of the family Anaplasmataceae in ticks in North Africa. A total of 418 questing Ixodes ricinus ticks collected in Tunisia and Morocco, as well as 188 Rhipicephalus ticks from dogs and 52 Hyalomma ticks from bovines in Tunisia, were included in this study. Of 324 adult I. ricinus ticks, 16.3% were positive for Ehrlichia spp., whereas only 3.4 and 2.8% of nymphs and larvae, respectively, were positive. A large heterogeneity was observed in the nucleotide sequences. Partial sequences identical to that of the agent of human granulocytic ehrlichiosis (HGE) were detected in I. ricinus and Hyalomma detritum, whereas partial sequences identical to that of Anaplasma platys were detected in Rhipicephalus sanguineus. However, variants of Anaplasma, provisionally designated Anaplasma-like, were predominant in the I. ricinus tick population in Maghreb. Otherwise, two variants of the genus Ehrlichia were detected in I. ricinus and H. detritum. Surprisingly, a variant of Wolbachia pipientis was evidenced from I. ricinus in Morocco. These results emphasized the potential risk of tick bites for human and animal populations in North Africa.}, } @article {pmid15734697, year = {2005}, author = {Haine, ER and Cook, JM}, title = {Convergent incidences of Wolbachia infection in fig wasp communities from two continents.}, journal = {Proceedings. Biological sciences}, volume = {272}, number = {1561}, pages = {421-429}, pmid = {15734697}, issn = {0962-8452}, mesh = {Animals ; Australia ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; Bayes Theorem ; Cytochromes b/genetics ; Ficus ; Models, Genetic ; Molecular Sequence Data ; Panama ; *Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Wasps/*microbiology ; Wolbachia/*genetics ; }, abstract = {Wide surveys suggest that Wolbachia bacteria infect ca. 20% of all insect species, but particular taxonomic or ecological groups may display significantly higher or lower incidences. We studied 61 fig wasp species in Australia and found the highest known incidence (67%) of infection in a targeted study of this nature. A comparable study in Panama reported a similar figure (59%), confirming the exceptionally high incidence of Wolbachia in fig wasps. Importantly, these are two independent estimates of Wolbachia incidence in fig wasp communities, because no host species, or even genera, are shared between localities. The high level of infection may reflect enhanced opportunities for horizontal transmission inside fig fruits. Although incidence was similar in Panama and Australia, the actual strains involved were different and region-specific. Local strains were shared by several host species, although there was often no obvious (direct) ecological link between two hosts with the same infection.}, } @article {pmid15710433, year = {2005}, author = {Mounsey, KE and Holt, DC and Fischer, K and Kemp, DJ and Currie, BJ and Walton, SF}, title = {Analysis of Sarcoptes scabiei finds no evidence of infection with Wolbachia.}, journal = {International journal for parasitology}, volume = {35}, number = {2}, pages = {131-135}, doi = {10.1016/j.ijpara.2004.11.007}, pmid = {15710433}, issn = {0020-7519}, mesh = {Animals ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/genetics ; Databases, Nucleic Acid ; Humans ; Polymerase Chain Reaction/methods ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae Infections/genetics/veterinary ; Sarcoptes scabiei/*genetics/parasitology ; Scabies/*genetics/parasitology ; Wolbachia/*genetics ; }, abstract = {The endosymbiont Wolbachia has been detected in a range of filarial nematodes and parasitic mites and is known to affect host reproductive compatibility and potentially evolutionary processes. PCR of Wolbachia surface protein (wsp), ftsZ and 16SrRNA genes from individual Sarcoptes scabiei mites obtained from a series of individual hosts, and database searches of an S. scabiei var. hominis EST library failed to detect Wolbachia genes. Therefore, Wolbachia appears not to be involved in the genetic subdivision observed between varieties of host-associated S. scabiei or, involved in the inflammatory disease pathogenesis of scabies unlike its activity in filarial infection.}, } @article {pmid15687015, year = {2005}, author = {Rao, RU and Atkinson, LJ and Vanderwall, RP and Weil, GJ}, title = {Brugia malayi: effects of gamma radiation on adult worms and their intracellular Wolbachia bacteria.}, journal = {Experimental parasitology}, volume = {109}, number = {2}, pages = {87-93}, doi = {10.1016/j.exppara.2004.11.009}, pmid = {15687015}, issn = {0014-4894}, mesh = {Aedes ; Animals ; Brugia malayi/embryology/microbiology/physiology/*radiation effects ; DNA, Bacterial/radiation effects ; DNA, Helminth/radiation effects ; Dose-Response Relationship, Radiation ; Embryonic Development/radiation effects ; Female ; *Gamma Rays ; Gerbillinae ; Male ; Microfilariae/radiation effects ; Microscopy, Electron ; Movement/radiation effects ; Reproduction/radiation effects ; Symbiosis ; Wolbachia/physiology/*radiation effects/ultrastructure ; }, abstract = {Prior studies have shown that intracellular Wolbachia endobacteria are necessary for the normal development, reproduction, and survival of filarial nematodes. The purpose of this study was to examine effects of gamma radiation on Wolbachia and reproduction in Brugia malayi adult worms. Worms were exposed to 0, 10, 25, 45, 75, and 105 krad of gamma radiation from a 137cesium source and cultured in vitro for 10 days. Irradiation reduced production of microfilariae in a dose-dependent manner. Embryograms of irradiated female worms showed dose-related abnormalities with arrested development at the early embryo stage. Irradiation reduced the viability of adult worms in a dose-dependent manner, but no lethal effect was observed. Electron microscopy studies showed that irradiation cleared Wolbachia from worm tissues. Real-time polymerase chain reaction studies demonstrated greatly reduced Wolbachia DNA in irradiated worms. These effects are essentially the same as those observed in adult worms treated with doxycycline. These studies suggest that effects of irradiation on reproduction in Brugia malayi may be caused by effects of irradiation on Wolbachia.}, } @article {pmid15681877, year = {2005}, author = {Rajan, TV}, title = {The eye does not see what the mind does not know: the bacterium in the worm.}, journal = {Perspectives in biology and medicine}, volume = {48}, number = {1}, pages = {31-41}, doi = {10.1353/pbm.2005.0015}, pmid = {15681877}, issn = {0031-5982}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Chlamydia/drug effects/isolation & purification/pathogenicity ; Culicidae/parasitology ; Elephantiasis, Filarial/drug therapy/*parasitology ; *Host-Parasite Interactions ; Humans ; India ; Molecular Biology ; Nematoda ; *Symbiosis ; United States ; Wolbachia/drug effects/isolation & purification/*pathogenicity ; Wuchereria bancrofti/microbiology/*parasitology ; }, abstract = {Symbiotic relationships underlie the evolutionary success of many different life forms. The filarial worms are long, slender nematode parasites that cause considerable pathology in large segments of the world's population. About 25 years ago, investigators first reported the presence of bacterial organisms living inside these parasitic worms. Recent molecular biological studies have indicated that these bacteria belong to the genus Wolbachia, members of which have been known to be associated with numerous species of insects. Elimination of the Wolbachia from the nematodes (by, for instance, treatment with broad spectrum antibiotics) results in profound disturbances in the physiology of the latter, including a complete block in reproduction. This observation, taken together with the fact that every individual worm examined to date contains Wolbachia, validates the classification of the latter as "endosymbionts." Many studies indicate that the Wolbachia may also play an important role in the pathology caused by the nematode worms, and that it might be possible to target therapy against the Wolbachia to treat the filarial disease. Intriguingly, the intense recent interest in Wolbachia is in complete contrast with the virtually complete indifference evoked by the original discovery of these organisms.}, } @article {pmid15674380, year = {2005}, author = {Reuter, M and Pedersen, JS and Keller, L}, title = {Loss of Wolbachia infection during colonisation in the invasive Argentine ant Linepithema humile.}, journal = {Heredity}, volume = {94}, number = {3}, pages = {364-369}, doi = {10.1038/sj.hdy.6800601}, pmid = {15674380}, issn = {0018-067X}, mesh = {Animals ; Ants/*genetics/*microbiology ; Argentina ; Environment ; *Phylogeny ; Population Dynamics ; *Selection, Genetic ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {WOLBACHIA are maternally inherited bacteria, which are very common in arthropods and nematodes. Wolbachia infection may affect host reproduction through feminisation, parthenogenesis, male-killing, cytoplasmic incompatibility and increased fecundity. Previous studies showing discrepancies between the phylogenies of Wolbachia and its arthropod hosts indicate that infection is frequently lost, but the causes of symbiont extinction have so far remained elusive. Here, we report data showing that colonisation of new habitats is a possible mechanism leading to the loss of infection. The presence and prevalence of Wolbachia were studied in three native and eight introduced populations of the Argentine ant Linepithema humile. The screening shows that the symbiont is common in the three native L. humile populations analysed. In contrast, Wolbachia was detected in only one of the introduced populations. The loss of infection associated with colonisation of new habitats may result from drift (founder effect) or altered selection pressures in the new habitat. Furthermore, a molecular phylogeny based on sequences of the Wolbachia wsp gene indicates that L. humile has been infected by a single strain. Horizontal transmission of the symbiont may be important in ants as suggested by the sequence similarity of strains in the three genera Linepithema, Acromyrmex, and Solenopsis native from South and Central America.}, } @article {pmid15660951, year = {2005}, author = {Malloch, G and Fenton, B}, title = {Super-infections of Wolbachia in byturid beetles and evidence for genetic transfer between A and B super-groups of Wolbachia.}, journal = {Molecular ecology}, volume = {14}, number = {2}, pages = {627-637}, doi = {10.1111/j.1365-294X.2005.02432.x}, pmid = {15660951}, issn = {0962-1083}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Base Sequence ; Cloning, Molecular ; Coleoptera/*genetics/*microbiology ; Cytoskeletal Proteins/genetics ; *Gene Transfer, Horizontal ; *Genetics, Population ; Likelihood Functions ; Models, Genetic ; Molecular Sequence Data ; *Phylogeny ; Restriction Mapping ; Sequence Alignment ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {Wolbachia are maternally inherited bacteria responsible for altering host reproduction. The two main groups found in insects, A and B, are based on molecular characterization using ribosomal, ftsZ, wsp (Wolbachia surface protein) or groE genes. We have used the wsp and ftsZ genes to study Wolbachia in byturid beetles. Byturus affinis contained a single copy of the ftsZ gene which grouped with A ftsZ sequences and a single copy of the wsp gene which grouped with B wsp sequences. This suggests that genetic exchange between A and B groups has occurred in the Wolbachia of this beetle. FtsZ and wsp sequences that were identical or nearly identical to those of B. affinis were found in B. tomentosus, suggesting that it also contains the same recombinant Wolbachia genotype. Most other byturids had more than one wsp sequence with at least one from the A and B groups, suggesting multiple copies of bacterial genes or multiple infections. B. ochraceus and B. unicolor both had four distinct wsp gene sequences. All the byturids had a closely related A wsp sequence and most a closely related B wsp sequence. Therefore, there appears to be an association between specific A and B wsp types.}, } @article {pmid15643971, year = {2005}, author = {Kyei-Poku, GK and Colwell, DD and Coghlin, P and Benkel, B and Floate, KD}, title = {On the ubiquity and phylogeny of Wolbachia in lice.}, journal = {Molecular ecology}, volume = {14}, number = {1}, pages = {285-294}, doi = {10.1111/j.1365-294X.2004.02409.x}, pmid = {15643971}, issn = {0962-1083}, mesh = {Animals ; Anoplura/*microbiology ; DNA Primers ; DNA, Bacterial/genetics/isolation & purification ; Humans ; Phthiraptera/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/*classification/*isolation & purification ; }, abstract = {Wolbachia are intracellular bacteria that occur in an estimated 20% of arthropod species. They are of broad interest because they profoundly affect the reproductive fitness of diverse host taxa. Here we document the apparent ubiquity and diversity of Wolbachia in the insect orders Anoplura (sucking lice) and Mallophaga (chewing lice), by detecting single or multiple infections in each of 25 tested populations of lice, representing 19 species from 15 genera spanning eight taxonomic families. Phylogenetic analyses indicate a high diversity of Wolbachia in lice, as evidenced by the identification of 39 unique strains. Some of these strains are apparently unique to lice, whereas others are similar to strains that infect other insect taxa. Wolbachia are transmitted from infected females to their offspring via egg cytoplasm, such that similar species of lice are predicted to have similar strains of Wolbachia. This predicted pattern is not supported in the current study and may reflect multiple events of recent horizontal transmission between host species. At present, there is no known mechanism that would allow for this latter mode of transmission to and within species of lice.}, } @article {pmid15612285, year = {2004}, author = {Marshall, JL}, title = {The Allonemobius-Wolbachia host-endosymbiont system: evidence for rapid speciation and against reproductive isolation driven by cytoplasmic incompatibility.}, journal = {Evolution; international journal of organic evolution}, volume = {58}, number = {11}, pages = {2409-2425}, doi = {10.1111/j.0014-3820.2004.tb00871.x}, pmid = {15612285}, issn = {0014-3820}, mesh = {Animals ; Base Sequence ; Cytoplasm/*microbiology ; DNA Primers ; DNA, Mitochondrial/genetics ; Fertilization/genetics ; *Genetic Variation ; Gryllidae/genetics/microbiology/*physiology ; Haplotypes/genetics ; Hybridization, Genetic ; Molecular Sequence Data ; *Phylogeny ; Reproduction/physiology ; Sequence Alignment ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; United States ; Wolbachia/*physiology ; }, abstract = {Evidence for the evolution of fertilization incompatibilities and rapid speciation can be biased by the occurrence of hybridization and reproductive endosymbionts such as Wolbachia. For example, patterns of mitochondrial DNA (mtDNA) variation can be obscured by mitotypes hitchhiking on extrachromosomal elements like Wolbachia, while such endosymbionts can also induce phenotypes that mirror the operation of intrinsic fertilization incompatibilities between species. Therefore, before strong inferences can be drawn concerning the rates and processes of speciation in arthropod systems, we must first assess whether extrinsic endosymbionts obscure patterns of speciation. Here, I use the Allonemobius fasciatus-socius species complex to determine what role Wolbachia has played in the presumed rapid divergence of this complex by analyzing patterns of mtDNA and nuclear DNA variation in conjunction with sequence and cytoplasmic incompatibility data on Wolbachia. Data on molecular variation suggest that Wolbachia has not induced a strong selective sweep of the mitochondrial genome; nor does Wolbachia appear to induce cytoplasmic incompatibility. Preliminary evidence indicates that a third species identified within this complex, A. sp. nov. Tex, is partially reproductively isolated from A. socius, its closest relative, via conspecific sperm precedence or some form of postzygotic isolation. Moreover, shared mitotypes between A. sp. nov. Tex and A. socius may indicate the occurrence of a hybrid zone between these species near the border of Texas and Louisiana, although they may also represent shared ancestral polymorphisms. Molecular data also indicate that all three species in this complex diverged from a common ancestor as recently as 3000-30,000 years ago. Finally, the radiation of this complex from its ancestral population likely occurred in the presence of one strain of Wolbachia, thus suggesting a minimal role for Wolbachia during this burst of speciation. In total, barriers to gene flow do appear to have evolved very rapidly in this group of crickets.}, } @article {pmid15611174, year = {2004}, author = {Shoemaker, DD and Dyer, KA and Ahrens, M and McAbee, K and Jaenike, J}, title = {Decreased diversity but increased substitution rate in host mtDNA as a consequence of Wolbachia endosymbiont infection.}, journal = {Genetics}, volume = {168}, number = {4}, pages = {2049-2058}, pmid = {15611174}, issn = {0016-6731}, mesh = {Animals ; *DNA, Mitochondrial ; Drosophila/*genetics/*microbiology ; *Evolution, Molecular ; Genetic Variation ; Mitochondria/genetics ; Molecular Sequence Data ; Wolbachia/*metabolism ; }, abstract = {A substantial fraction of insects and other terrestrial arthropods are infected with parasitic, maternally transmitted endosymbiotic bacteria that manipulate host reproduction. In addition to imposing direct selection on the host to resist these effects, endosymbionts may also have indirect effects on the evolution of the mtDNA with which they are cotransmitted. Patterns of mtDNA diversity and evolution were examined in Drosophila recens, which is infected with the endosymbiont Wolbachia, and its uninfected sister species D. subquinaria. The level of mitochondrial, but not nuclear, DNA diversity is much lower in D. recens than in D. subquinaria, consistent with the hypothesized diversity-purging effects of an evolutionarily recent Wolbachia sweep. The d(N)/d(S) ratio in mtDNA is significantly greater in D. recens, suggesting that Muller's ratchet has brought about an increased rate of substitution of slightly deleterious mutations. The data also reveal elevated rates of synonymous substitutions in D. recens, suggesting that these sites may experience weak selection. These findings show that maternally transmitted endosymbionts can severely depress levels of mtDNA diversity within an infected host species, while accelerating the rate of divergence among mtDNA lineages in different species.}, } @article {pmid15605657, year = {2004}, author = {Rasgon, JL and Scott, TW}, title = {Phylogenetic characterization of Wolbachia symbionts infecting Cimex lectularius L. and Oeciacus vicarius Horvath (Hemiptera: Cimicidae).}, journal = {Journal of medical entomology}, volume = {41}, number = {6}, pages = {1175-1178}, doi = {10.1603/0022-2585-41.6.1175}, pmid = {15605657}, issn = {0022-2585}, support = {GM20092/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bedbugs/*microbiology ; Cimicidae/*microbiology ; DNA Primers ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/*classification/genetics/*isolation & purification ; }, abstract = {Wolbachia symbionts are obligate intracellular bacteria that cause host reproductive alterations in many arthropods and filarial nematodes. We identified Wolbachia symbionts in the cliff swallow bug (Oeciacus vicarious Horvath) and the human bed bug (Cimex lectularius L.) (Hemiptera: Cimicidae) by polymerase chain reaction (PCR) amplification and sequencing using Wolbachia-specific 16S rDNA and FtsZ primers. Phylogenetic analyses using Bayesian, maximum likelihood, and maximum parsimony algorithms indicated, with strong support, that (1) Wolbachia infections in these two cimicid hosts form a monophyletic group, and (2) the Wolbachia strains detected belong to the F clade, previously associated with termites, weevils, and filarial nematodes.}, } @article {pmid15579697, year = {2004}, author = {Dyer, KA and Jaenike, J}, title = {Evolutionarily stable infection by a male-killing endosymbiont in Drosophila innubila: molecular evidence from the host and parasite genomes.}, journal = {Genetics}, volume = {168}, number = {3}, pages = {1443-1455}, pmid = {15579697}, issn = {0016-6731}, mesh = {Animals ; Drosophila/genetics/*microbiology ; Evolution, Molecular ; Female ; Genetic Variation ; Male ; Molecular Sequence Data ; Phylogeny ; Sex Ratio ; Wolbachia/genetics/*physiology ; }, abstract = {Maternally inherited microbes that spread via male-killing are common pathogens of insects, yet very little is known about the evolutionary duration of these associations. The few examples to date indicate very recent, and thus potentially transient, infections. A male-killing strain of Wolbachia has recently been discovered in natural populations of Drosophila innubila. The population-level effects of this infection are significant: approximately 35% of females are infected, infected females produce very strongly female-biased sex ratios, and the resulting population-level sex ratio is significantly female biased. Using data on infection prevalence and Wolbachia transmission rates, infected cytoplasmic lineages are estimated to experience a approximately 5% selective advantage relative to uninfected lineages. The evolutionary history of this infection was explored by surveying patterns of polymorphism in both the host and parasite genomes, comparing the Wolbachia wsp gene and the host mtDNA COI gene to five host nuclear genes. Molecular data suggest that this male-killing infection is evolutionarily old, a conclusion supported with a simple model of parasite and mtDNA transmission dynamics. Despite a large effective population size of the host species and strong selection to evolve resistance, the D. innubila-Wolbachia association is likely at a stable equilibrium that is maintained by imperfect maternal transmission of the bacteria rather than partial resistance in the host species.}, } @article {pmid15578046, year = {2005}, author = {Gotoh, T and Noda, H and Fujita, T and Iwadate, K and Higo, Y and Saito, S and Ohtsuka, S}, title = {Wolbachia and nuclear-nuclear interactions contribute to reproductive incompatibility in the spider mite Panonychus mori (Acari: Tetranychidae).}, journal = {Heredity}, volume = {94}, number = {2}, pages = {237-246}, doi = {10.1038/sj.hdy.6800605}, pmid = {15578046}, issn = {0018-067X}, mesh = {Analysis of Variance ; Animals ; Cell Nucleus/*genetics ; Crosses, Genetic ; Cytoplasm/microbiology ; DNA Primers ; Female ; Geography ; Japan ; Male ; Polymerase Chain Reaction ; Reproduction/genetics/physiology ; Rifampin ; *Symbiosis ; Tetranychidae/genetics/*microbiology/*physiology ; *Wolbachia ; }, abstract = {Maternally transmitted bacteria of the genus Wolbachia are obligate, intracellular symbionts that are responsible for cytoplasmic incompatibility in a wide range of arthropods such as insects and mites. Spider mites often show uni- and bidirectional incompatibilities among populations with and without Wolbachia. Therefore, we surveyed the presence of Wolbachia by PCR and then conducted crossing experiments among 25 populations of Panonychus mori to determine how Wolbachia are related to the incompatibility in this species. Five out of the 25 populations were infected with Wolbachia. These five populations were treated with an antibiotic (rifampicin) to eliminate Wolbachia. We carried out round-robin crosses among 20 Wolbachia-uninfected populations, five infected populations and five rifampicin-treated populations (30 x 30=900 crosses in total). Incompatibility among P. mori populations was caused by Wolbachia infection, nuclear-cytoplasmic interactions or nuclear-nuclear interactions. Wolbachia-mediated incompatibility was observed in crosses between uninfected females and infected males or between females and males harboring different Wolbachia strains. Nuclear-cytoplasmic interactions may be responsible for the unidirectional incompatibility in crosses between the two northern populations and one of the southern populations. Bidirectional incompatibility caused by nuclear-nuclear interactions was observed in 99 combinations of interpopulation crosses (99/300=0.33). Although no geographical trends were detected in the distribution of bidirectionally compatible populations, the results reveal a genetic divergence among P. mori populations.}, } @article {pmid15571631, year = {2004}, author = {Veneti, Z and Toda, MJ and Hurst, GD}, title = {Host resistance does not explain variation in incidence of male-killing bacteria in Drosophila bifasciata.}, journal = {BMC evolutionary biology}, volume = {4}, number = {}, pages = {52}, pmid = {15571631}, issn = {1471-2148}, mesh = {Animals ; Animals, Wild/*genetics/*microbiology ; Breeding ; Crosses, Genetic ; Cytoplasm/genetics/microbiology ; Drosophila/*genetics/*microbiology ; Female ; Feminization/genetics ; Genetic Variation/*genetics ; Gram-Negative Bacterial Infections/*genetics/*mortality/transmission ; Immunity, Innate/*genetics ; Incidence ; Infectious Disease Transmission, Vertical ; Japan ; Male ; Phenotype ; Prevalence ; Sex Factors ; *Sex Ratio ; Temperature ; Wolbachia/*pathogenicity ; }, abstract = {BACKGROUND: Selfish genetic elements that distort the sex ratio are found widely. Notwithstanding the number of records of sex ratio distorters, their incidence is poorly understood. Two factors can prevent a sex ratio distorter from invading: inability of the sex ratio distorter to function (failure of mechanism or transmission), and lack of drive if they do function (inappropriate ecology for invasion). There has been no test to date on factors causing variation in the incidence of sex ratio distorting cytoplasmic bacteria. We therefore examined whether absence of the male-killing Wolbachia infection in D. bifasciata in Hokkaido island of Japan, in contrast to the presence of infection on the proximal island of Honshu, was associated with failure of the infection to function properly on the Hokkaido genetic background.

RESULTS: The male-killer both transmitted and functioned well following introgression to each of 24 independent isofemale inbred lines carrying Hokkaido genetic backgrounds. This was maintained even under stringent conditions of temperature. We therefore reject the hypothesis that absence of infection is due to its inability to kill males and transmit on the Hokkaido genetic background. Further trap data indicates that D. bifasciata may occur at different densities in Hokkaido and Honshu populations, giving some credence to the idea that ecological differentiation could be important.

CONCLUSIONS: The absence of the infection from the Hokkaido population is not caused by failure of the male-killer to function on the Hokkaido genetic background.}, } @article {pmid15567711, year = {2004}, author = {Cho, KO}, title = {Wolbachia bacteria, the cause for false vesicular staining pattern in Drosophila melanogaster.}, journal = {Gene expression patterns : GEP}, volume = {5}, number = {2}, pages = {167-170}, doi = {10.1016/j.modgep.2004.08.008}, pmid = {15567711}, issn = {1567-133X}, support = {R29 NS35532/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/*immunology/metabolism ; Cytoplasmic Vesicles/*metabolism ; Drosophila Proteins/*immunology/metabolism ; Drosophila melanogaster/metabolism/*microbiology/ultrastructure ; Immune Sera/*immunology ; Immunohistochemistry ; Wolbachia/*immunology ; }, abstract = {Majority of fly laboratory strains is infected with Wolbachia, intracellular rickettsial-type symbiotic bacteria widespread in various organisms including insects and nematodes. To make the matter worse, I found that certain antisera used for fly immunocytochemistry can recognize Wolbachia bacteria in addition to their own antigens, due to impurity in the antisera generated against the recombinant fusion proteins frequently used as antigens. Thus, combinatorial use of contaminated antisera and Wolbachia-infected flies can result in serious misinterpretations, which can be avoided by curing laboratory strains of Wolbachia.}, } @article {pmid15562699, year = {2004}, author = {Engelstädter, J and Montenegro, H and Hurst, GD}, title = {To what extent do different types of sex ratio distorters interfere?.}, journal = {Evolution; international journal of organic evolution}, volume = {58}, number = {10}, pages = {2382-2386}, doi = {10.1111/j.0014-3820.2004.tb01612.x}, pmid = {15562699}, issn = {0014-3820}, mesh = {Animals ; Computer Simulation ; Diptera/*genetics/microbiology ; Meiosis/genetics ; *Models, Genetic ; Repetitive Sequences, Nucleic Acid/genetics ; *Sex Ratio ; Wolbachia/*physiology ; X Chromosome/*genetics ; }, abstract = {Within the Diptera, two different selfish genetic elements are known to cause the production of female-biased sex ratios: maternally inherited bacteria that kill male zygotes (male-killers), and X chromosomes causing the degeneration of Y-bearing sperm in males (meiotic drive). We here develop a mathematical model for the dynamics of these two sex-ratio distorters where they co-occur. We show that X chromosome meiotic drive elements can be expected to substantially lower the equilibrium frequency of male-killers and can even lead to their extinction. Conversely, male-killers can also decrease the equilibrium frequency of X drivers and cause their extinction. Thus, we predict that there will be some complementarity in the incidence of X chromosome meiotic drive and male-killing in natural populations, with a lower than expected number of species bearing both elements.}, } @article {pmid15562683, year = {2004}, author = {Dedeine, F and Vavre, F and Shoemaker, DD and Boulétreau, M}, title = {Intra-individual coexistence of a Wolbachia strain required for host oogenesis with two strains inducing cytoplasmic incompatibility in the wasp Asobara tabida.}, journal = {Evolution; international journal of organic evolution}, volume = {58}, number = {10}, pages = {2167-2174}, doi = {10.1111/j.0014-3820.2004.tb01595.x}, pmid = {15562683}, issn = {0014-3820}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm/microbiology ; Oogenesis/*physiology ; Polymerase Chain Reaction ; Reproduction/physiology ; Species Specificity ; *Symbiosis ; Wasps/*microbiology/physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Cytoplasmically inherited symbiotic Wolbachia bacteria are known to induce a diversity of phenotypes on their numerous arthropod hosts including cytoplasmic incompatibility, male-killing, thelytokous parthenogenesis, and feminization. In the wasp Asobara tabida (Braconidae), in which all individuals harbor three genotypic Wolbachia strains (wAtab1, wAtab2 and wAtab3), the presence of Wolbachia is required for insect oogenesis. To elucidate the phenotype of each Wolbachia strain on host reproduction, especially on oogenesis, we established lines of A. tabida harboring different combinations of these three bacterial strains. We found that wAtab3 is essential for wasp oogenesis, whereas the two other strains, wAtabl and wAtab2, seem incapable to act on this function. Furthermore, interline crosses showed that strains wAtab1 and wAtab2 induce partial (about 78%) cytoplasmic incompatibility of the female mortality type. These results support the idea that bacterial genotype is a major factor determining the phenotype induced by Wolbachia on A. tabida hosts. We discuss the implications of these findings for current hypotheses regarding the evolutionary mechanisms by which females of A. tabida have become dependent on Wolbachia for oogenesis.}, } @article {pmid15562682, year = {2004}, author = {Dobson, SL}, title = {Evolution of Wolbachia cytoplasmic incompatibility types.}, journal = {Evolution; international journal of organic evolution}, volume = {58}, number = {10}, pages = {2156-2166}, doi = {10.1111/j.0014-3820.2004.tb01594.x}, pmid = {15562682}, issn = {0014-3820}, mesh = {Animals ; *Biological Evolution ; Cytoplasm/microbiology ; Insecta/*microbiology ; *Models, Biological ; Mutation/genetics ; Population Dynamics ; Reproduction/genetics ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {The success of obligate endosymbiotic Wolbachia infections in insects is due in part to cytoplasmic incompatibility (CI), whereby Wolbachia bacteria manipulate host reproduction to promote their invasion and persistence within insect populations. The observed diversity of CI types raises the question of what the evolutionary pathways are by which a new CI type can evolve from an ancestral type. Prior evolutionary models assume that Wolbachia exists within a host individual as a clonal infection. While endosymbiotic theory predicts a general trend toward clonality, Wolbachia provides an exception in which there is selection to maintain diversity. Here, evolutionary trajectories are discussed that assume that a novel Wolbachia variant will co-exist with the original infection type within a host individual as a superinfection. Relative to prior models, this assumption relaxes requirements and allows additional pathways for the evolution of novel CI types. In addition to describing changes in the Wolbachia infection frequency associated with the hypothesized evolutionary events, the predicted impact of novel CI variants on the host population is also described. This impact, resulting from discordant evolutionary interests of symbiont and host, is discussed as a possible cause of Wolbachia loss from the host population or host population extinction. The latter is also discussed as the basis for an applied strategy for the suppression of insect pest populations. Model predictions are discussed relative to a recently published Wolbachia genome sequence and prior characterization of CI in naturally and artificially infected insects.}, } @article {pmid15562592, year = {2004}, author = {Dittmar, K and Whiting, MF}, title = {New Wolbachia endosymbionts from Nearctic and Neotropical fleas (Siphonaptera).}, journal = {The Journal of parasitology}, volume = {90}, number = {5}, pages = {953-957}, doi = {10.1645/GE-186R}, pmid = {15562592}, issn = {0022-3395}, mesh = {Animals ; Base Sequence ; Cloning, Molecular ; Consensus Sequence ; DNA, Bacterial/chemistry ; DNA, Ribosomal/*chemistry ; Female ; Male ; Molecular Sequence Data ; North America ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/*genetics ; Siphonaptera/*microbiology ; South America ; Symbiosis ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {Several species of fleas (Siphonaptera), ectoparasites of mammals and birds, have recently been shown to harbor species of Wolbachia. Here, we extend this data set to 20 more species of Siphonaptera (Rhopalopsyllidae, Stephanocircidae, Pulicidae, Ceratophyllidae, Ctenophthalmidae, Ischnopsyllidae, Leptopsyllidae, and Malacopsyllidae) from sylvatic populations throughout the Nearctic and Neotropical regions. Using polymerase chain reaction, we targeted the Wolbachia 16S ribosomal DNA (rDNA) gene. Applying maximum parsimony- and maximum likelihood-based algorithms, as well as statistical parsimony, we conducted a phylogenetic analysis of Wolbachia 16S rDNA to evaluate its position within the known Wolbachia spp. The analysis recovered the siphonapteran Wolbachia 16S rDNA sequences as a monophyletic group and shows multiple haplotype connections between the Neotropical and Nearctic Wolbachia strains of fleas.}, } @article {pmid15562287, year = {2005}, author = {Mouton, L and Henri, H and Boulétreau, M and Vavre, F}, title = {Multiple infections and diversity of cytoplasmic incompatibility in a haplodiploid species.}, journal = {Heredity}, volume = {94}, number = {2}, pages = {187-192}, doi = {10.1038/sj.hdy.6800596}, pmid = {15562287}, issn = {0018-067X}, mesh = {Animals ; Diploidy ; Female ; France ; Haploidy ; Male ; Reproduction/physiology ; Species Specificity ; Wasps/*microbiology/*physiology ; *Wolbachia ; }, abstract = {Cytoplasmic incompatibility (CI) is a sperm-egg incompatibility commonly induced by the intracellular endosymbiont bacterium Wolbachia that, in diploid species, results in embryo mortality. In haplodiploid species, two types of CI exist depending on whether the incompatible fertilized eggs develop into males (male development (MD)) or abort (female mortality (FM)). CI allows multiple infections to be maintained in host populations, and thus allows interactions to occur between co-infecting strains. In Leptopilina heterotoma, three Wolbachia strains coexist naturally (wLhet1, wLhet2, wLhet3). When these three strains are all present, they induce a CI of FM type, whereas wLhet1 alone expresses a CI phenotype intermediate between MD and FM. Here, we compare CI effects in crosses involving insect lines sharing the same nuclear background, but harboring different mixtures of strains. Mating experiments showed that: (i) wLhet2 and wLhet3 also induce an intermediate CI when acting alone, and show a bidirectional incompatibility; (ii) there is no interaction between the co-infecting strains in CI expression; (iii) the diversity of Wolbachia present within a male host influences the expression of CI: an increase in the number of strains is correlated with a decrease in the proportion of the MD type, which is also correlated with an increase in bacterial density. All these data suggest that the CI of FM type results from a stronger effect than the MD type, which conflicts with the conventional hypotheses used to explain CI diversity in haplodiploids, and could provide some new information about CI mechanisms in insects.}, } @article {pmid15535618, year = {2004}, author = {Dean, JL and Dobson, SL}, title = {Characterization of Wolbachia infections and interspecific crosses of Aedes (Stegomyia) polynesiensis and Ae. (Stegomyia) riversi (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {41}, number = {5}, pages = {894-900}, doi = {10.1603/0022-2585-41.5.894}, pmid = {15535618}, issn = {0022-2585}, support = {AI51533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/classification/*microbiology ; Animals ; Crosses, Genetic ; Female ; Fertility ; Male ; Oviposition ; Phylogeny ; Polymerase Chain Reaction ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Prior studies have identified a complicated pattern of interspecific hybridization between members of the Aedes (Stegomyia) scutellaris (Walker) mosquito group, which includes medically important vectors of bancroftian filariasis and dengue. Here, we report that two members of the group, Aedes polynesiensis Marks and Aedes riversi Bohart & Ingram, are both infected with intracellular Wolbachia bacteria. Sequencing of the Wolbachia wsp gene demonstrates that the infections differ from each other and from Wolbachia infections previously reported in mosquitoes. Aedes polynesiensis is the first mosquito identified with a wMel Wolbachia type. Intraspecific crosses of infected and aposymbiotic lines generated via antibiotic treatment show that the Wolbachia infections in both species cause high levels of cytoplasmic incompatibility. Interspecific crosses show that the two species are reproductively isolated. However, repeating the interspecific crosses with aposymbiotic mosquito strains demonstrates that the Wolbachia infections play a role in preventing hybrid offspring. We discuss Wolbachia infections in relation to better defining the evolutionary relationships and causes of speciation within the group, understanding the basis for the observed east-to-west gradient in filarial refractoriness, and developing novel genetic control measures.}, } @article {pmid15535575, year = {2000}, author = {Kittayapong, P and Baisley, KJ and Baimai, V and O'Neill, SL}, title = {Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {37}, number = {3}, pages = {340-345}, doi = {10.1093/jmedent/37.3.340}, pmid = {15535575}, issn = {0022-2585}, support = {G0700837/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/microbiology ; Animals ; Asia, Southeastern ; Culicidae/*microbiology ; Polymerase Chain Reaction ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia are a group of intracellular inherited bacteria that infect a wide range of arthropods. They are associated with a variety of reproductive alterations in their hosts, the best known being cytoplasmic incompatability. The Wolbachia pipientis assemblage has been divided into two major groups (A and B) and 12 subgroups. We report herein the first systematic survey of Wolbachia in mosquitoes, and the first survey classifying Wolbachia infections by subgroup. Wolbachia were detected in 28.1% of 89 wild-caught mosquito species, based on a polymerase chain reaction assay using ftsZ and wsp gene primers. Infections were found in all major disease vector genera except Anopheles. Nine of the 12 Wolbachia subgroups were represented. Group B Wolbachia strains showed more phylogenetic concordance with their host taxa than group A strains. Of the 25 positive mosquito species, five were superinfected with group A bacteria strains (AA), eight were superinfected with A and B strains (AB), and one was superinfected with group B strains (BB). The widespread distribution of Wolbachia among mosquito species further supports their potential importance in the genetic control of disease vectors.}, } @article {pmid15533291, year = {2004}, author = {Heukelbach, J and Bonow, I and Witt, L and Feldmeier, H and Fischer, P}, title = {High infection rate of Wolbachia endobacteria in the sand flea Tunga penetrans from Brazil.}, journal = {Acta tropica}, volume = {92}, number = {3}, pages = {225-230}, doi = {10.1016/j.actatropica.2004.08.005}, pmid = {15533291}, issn = {0001-706X}, mesh = {Animals ; Brazil ; Feces/microbiology ; Female ; Male ; Oocytes/microbiology ; Ovary/microbiology ; Siphonaptera/*microbiology ; Staining and Labeling ; Wolbachia/*isolation & purification ; }, abstract = {Tunga penetrans is an ectoparasite causing considerable morbidity in endemic communities. Recently, endobacteria of the genus Wolbachia were identified also in T. penetrans. Since Wolbachia were suggested as targets for intervention of insect pests and human filariasis, sand fleas were collected from infested humans, dogs and rats in a hyperendemic area in northeastern Brazil, and screened for Wolbachia infections. Twenty-one adult fleas and four batches of flea eggs were examined by PCR using primers targeting the 16S rDNA, the DNA coding for FtsZ cell-cycle protein or a Wolbachia surface protein (WSP-1). Wolbachia were detected in all examined samples from eggs, free-living male and female fleas and from neosomic female fleas. No Wolbachia DNA was detected in two samples containing flea faeces. In addition, Wolbachia were labelled by immunohistology in the ovaries of 37 female fleas using antisera raised against WSP-1 of Wolbachia the filarial parasite Dirofilaria immitis. In the vicinity of the embedded fleas containing the Wolbachia, infiltrations of neutrophils and macrophages were observed. This study showed that Wolbachia endobacteria are abundant in T. penetrans and that all examined fleas were infected by these endobacteria. Our findings may have important implications for the future development of control strategies for human tungiasis.}, } @article {pmid15523503, year = {2005}, author = {Jeong, G and Stouthamer, R}, title = {Genetics of female functional virginity in the parthenogenesis-Wolbachia infected parasitoid wasp Telenomus nawai (Hymenoptera: Scelionidae).}, journal = {Heredity}, volume = {94}, number = {4}, pages = {402-407}, doi = {10.1038/sj.hdy.6800617}, pmid = {15523503}, issn = {0018-067X}, mesh = {Animals ; Female ; *Genes, Recessive ; Male ; *Mutation ; Parthenogenesis/*genetics/physiology ; Reproduction/genetics ; Wasps/*genetics/microbiology ; *Wolbachia ; }, abstract = {A lepidopteran egg parasitoid species Telenomus nawai consists of two distinct populations with different reproductive modes. One is a completely thelytokous population consisting of females only, whereas the other displays arrhenotokous reproduction where fertilized eggs develop into diploid females and unfertilized eggs into haploid males. Thelytoky in T. nawai is caused by a bacterial symbiont, the parthenogenesis-inducing (PI) Wolbachia. Recent theoretical studies have shown that when a PI-Wolbachia is spreading in a population, mutations that allow uninfected females to produce more male offspring will spread rapidly eventually becoming fixed. The consequence of such a mutation is that sexual reproduction is no longer successful in infected females. Here we determine the genetic basis of the females' inability to reproduce sexually by introgressing the genome of a thelytokous line into an arrhenotokous line. The results suggest that the mutations are recessive and inherited either as a single-locus major gene with some modifiers, or as two partially linked loci.}, } @article {pmid15521450, year = {2004}, author = {Charlat, S and Riegler, M and Baures, I and Poinsot, D and Stauffer, C and Merçot, H}, title = {Incipient evolution of Wolbachia compatibility types.}, journal = {Evolution; international journal of organic evolution}, volume = {58}, number = {9}, pages = {1901-1908}, doi = {10.1111/j.0014-3820.2004.tb00478.x}, pmid = {15521450}, issn = {0014-3820}, mesh = {Animals ; *Biological Evolution ; Crosses, Genetic ; Cytoplasm/microbiology/physiology ; Drosophila/*microbiology/physiology ; Host-Parasite Interactions ; *Models, Biological ; Reproduction/genetics/physiology ; Species Specificity ; Statistics, Nonparametric ; Tephritidae/*microbiology/physiology ; Wolbachia/*genetics/physiology ; }, abstract = {Cytoplasmic incompatibility (CI) is induced in arthropods by the maternally inherited bacterium Wolbachia. When infected males mate with uninfected females or with females bearing a different Wolbachia variant, paternal chromosomes behave abnormally and embryos die. This pattern can be interpreted as resulting from two bacterial effects: One (usually termed mod, for modification) would affect sperm and induce embryo death, unless Wolbachia is also present in the egg, which implies the existence of a second effect, usually termed resc, for rescue. The fact that CI can occur in crosses between males and females infected by different Wolbachia shows that mod and resc interact in a specific manner. In other words, different compatibility types, or mod/resc pairs seem to have diverged from one (or a few) common ancestor(s). We are interested in the process allowing the evolution of mod/resc pairs. Here this question is addressed experimentally after cytoplasmic injection into a single host species (Drosophila simulans) by investigating compatibility relationships between closely related Wolbachia variants naturally evolving in different dipteran hosts: D. simulans, Drosophila melanogaster, and Rhagoletis cerasi. Our results suggest that closely related bacteria can be totally or partially incompatible. The compatibility relationships observed can be explained using a formal description of the mod and resc functions, implying both qualitative and quantitative variations.}, } @article {pmid15501467, year = {2004}, author = {Engelstädter, J and Telschow, A and Hammerstein, P}, title = {Infection dynamics of different Wolbachia-types within one host population.}, journal = {Journal of theoretical biology}, volume = {231}, number = {3}, pages = {345-355}, doi = {10.1016/j.jtbi.2004.06.029}, pmid = {15501467}, issn = {0022-5193}, mesh = {Animals ; Cytoplasm/parasitology ; Death ; Female ; Host-Parasite Interactions ; Male ; Models, Biological ; Parthenogenesis ; Rickettsiaceae Infections/*parasitology ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia are widespread intracellular symbionts of arthropods which are known to cause several reproductive manipulations in their hosts, the commonest of which being cytoplasmic incompatibility (CI), male killing (MK), and the induction of parthenogenesis (PI). Strains of endosymbionts inducing one of these effects can be referred to as 'Wolbachia-types'. Here, we try to ascertain whether two of these Wolbachia-types can stably coexist within one population. We investigate this question by means of two discrete-time mathematical models which describe the dynamics of an infection of a host population with either CI- and MK- or CI- and PI-Wolbachia. We derive analytical solutions for two special cases of each model showing that stable coexistence of the respective Wolbachia-types is not possible if no doubly infected individuals occur within the population and that stable coexistence is possible when doubly infected hosts do exist and transmission of the endosymbionts is perfect. Moreover, we show that a population infected with either CI- or MK-Wolbachia at equilibrium can resist invasion of the respective other Wolbachia-type as a single infection. In contrast, a population infected with CI-Wolbachia can be invaded by PI-Wolbachia as a single infection with the CI-Wolbachia going extinct. Computer simulations confirmed these findings for the general models. We discuss our results with respect to the prevalence of the Wolbachia-types considered here and the emergence of PI- from CI-Wolbachia.}, } @article {pmid15491598, year = {2004}, author = {Harcombe, W and Hoffmann, AA}, title = {Wolbachia effects in Drosophila melanogaster: in search of fitness benefits.}, journal = {Journal of invertebrate pathology}, volume = {87}, number = {1}, pages = {45-50}, doi = {10.1016/j.jip.2004.07.003}, pmid = {15491598}, issn = {0022-2011}, mesh = {Animals ; Drosophila melanogaster/*microbiology/*physiology ; Heat Stress Disorders ; Host-Parasite Interactions ; Nutritional Physiological Phenomena ; Rickettsiaceae Infections/*physiopathology ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {Insect endosymbionts often influence host nutrition but these effects have not been comprehensively investigated in Wolbachia endosymbionts that are widespread in insects. Using strains of Drosophila melanogaster with the wMel Wolbachia infection, we showed that Wolbachia did not influence adult starvation resistance. Wolbachia also had no effect on larval development time or the size of emerging adults from a low nutrition medium. While Wolbachia may influence the expression of heat shock proteins, we found that there was no effect on adult heat resistance when tested in terms of survival or virility following heat stress. The absence of nutrition or stress effects suggests that other processes maintain wMel frequencies in natural populations of Drosophila melanogaster.}, } @article {pmid15491544, year = {2004}, author = {Luchetti, A and Mantovani, B and Fioravanti, ML and Trentini, M}, title = {Wolbachia infection in the newly described Ecuadorian sand flea, Tunga trimamillata.}, journal = {Experimental parasitology}, volume = {108}, number = {1-2}, pages = {18-23}, doi = {10.1016/j.exppara.2004.07.003}, pmid = {15491544}, issn = {0014-4894}, mesh = {Animals ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/*analysis ; DNA, Ribosomal/*analysis ; Female ; Genotype ; Host-Parasite Interactions ; Male ; Molecular Sequence Data ; Phenotype ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/*genetics ; Reproduction ; Sequence Alignment ; Sex Ratio ; Siphonaptera/genetics/*microbiology/physiology ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Wolbachia pipientis is an intracellular endosymbiont producing reproductive alterations in its hosts. This bacterium have been reported in many arthropods and nematodes. By PCR amplification and sequencing of the 16S rDNA and ftsZ genes we have identified a Wolbachia strain in the newly described sand-flea, Tunga trimamillata. Prevalence of this endosymbiont in the 26 individuals screened is equal to 35%. Sympatric and allopatric specimens of the related species Tunga penetrans were also analysed, but in contrast to literature data, Wolbachia appears absent in the presently analysed 24 specimens. Field studies evidence a female-biased sex-ratio in T. trimamillata, suggesting that Wolbachia may cause sex-ratio distortion in this species. By means of BLAST search and phylogenetic analysis we found that the Wolbachia strain from T. trimamillata pertains to the arthropod-infecting Wolbachia; this strain is highly differentiated from the Wolbachia strain of T. penetrans described in literature.}, } @article {pmid15482887, year = {2004}, author = {Morchón, R and Ferreira, AC and Martín-Pacho, JR and Montoya, A and Mortarino, M and Genchi, C and Simón, F}, title = {Specific IgG antibody response against antigens of Dirofilaria immitis and its Wolbachia endosymbiont bacterium in cats with natural and experimental infections.}, journal = {Veterinary parasitology}, volume = {125}, number = {3-4}, pages = {313-321}, doi = {10.1016/j.vetpar.2004.08.003}, pmid = {15482887}, issn = {0304-4017}, mesh = {Animals ; Anthelmintics/therapeutic use ; Antibodies, Bacterial/blood ; Antibodies, Helminth/blood ; Antibody Specificity ; Antigens, Helminth/*immunology ; Cat Diseases/drug therapy/immunology/microbiology/*parasitology ; Cats ; Dirofilaria immitis/*immunology/metabolism ; Dirofilariasis/drug therapy/*immunology/parasitology ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Heart/parasitology ; Immunoglobulin G/*biosynthesis/blood/immunology ; Ivermectin/therapeutic use ; Male ; Peptide Fragments/immunology ; Pulmonary Artery/parasitology ; Wolbachia/*immunology/metabolism ; }, abstract = {Sera from three groups of cats under different experimental conditions were studied by ELISA to assess the host's immune response against synthetic peptides derived from Dirofilaria immitis (Dipp) and against the surface protein of its endosymbiont, Wolbachia (WSPr). In experimentally infected cats (Group 1), an increase of IgG antibody against both Dipp and WSPr was observed from 2 months post-infection until the end of the study, 6 months post-infection. In experimentally infected cats, treated against infective larvae (Group 2), anti-Dipp IgG decreased dramatically from 4 months post-infection (3 months post treatment), showing very low values till the end of the study (6.5 months from infection, 5.5 months from treatment), while anti-WSP IgG increased constantly till the end of the study. Of 49 outdoor, asymptomatic cats exposed to a high risk of natural infection (Group 3), 9 were positive for anti-Dipp IgG and for a validated, in-clinic commercial antibody diagnostic kit for cats. Two cats were also found positive for circulating antigens of adult female worm. Anti-WSPr IgG were found in five of nine anti-Dipp IgG-positive sera and from eight ELISADipp-negative sera. Our results confirm the strong IgG response in heartworm infected cats and demonstrate the involvement of the Wolbachia endosymbiont in the immune reaction to the parasite both in experimentally infected cats and in cats exposed to a high risk of natural infection.}, } @article {pmid15469918, year = {2004}, author = {Zabalou, S and Riegler, M and Theodorakopoulou, M and Stauffer, C and Savakis, C and Bourtzis, K}, title = {Wolbachia-induced cytoplasmic incompatibility as a means for insect pest population control.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {101}, number = {42}, pages = {15042-15045}, pmid = {15469918}, issn = {0027-8424}, mesh = {Animals ; Ceratitis capitata/*microbiology ; Cytoplasm/microbiology ; Female ; Male ; Pest Control, Biological/*methods ; Tephritidae/microbiology ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations. Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents. Wolbachia manipulate host reproduction by using several strategies, one of which is cytoplasmic incompatibility (CI) [Stouthamer, R., Breeuwer, J. A. J. & Hurst, G. D. D. (1999) Annu. Rev. Microbiol. 53, 71-102]. We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor. Wolbachia induced complete CI in the novel host. Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced CI could be used as a novel environmentally friendly tool for the control of medfly populations. The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations.}, } @article {pmid15454547, year = {2004}, author = {Pannebakker, BA and Beukeboom, LW and van Alphen, JJ and Brakefield, PM and Zwaan, BJ}, title = {The genetic basis of male fertility in relation to haplodiploid reproduction in Leptopilina clavipes (Hymenoptera: Figitidae).}, journal = {Genetics}, volume = {168}, number = {1}, pages = {341-349}, pmid = {15454547}, issn = {0016-6731}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Chromosome Mapping ; Crosses, Genetic ; DNA Primers ; Female ; Fertility/genetics/physiology ; Male ; Nucleic Acid Amplification Techniques ; Parthenogenesis/*genetics ; *Phenotype ; Polymorphism, Restriction Fragment Length ; Quantitative Trait Loci ; Reproduction/physiology ; *Selection, Genetic ; Wasps/*genetics/*microbiology/physiology ; Wolbachia/drug effects/*physiology ; }, abstract = {Traits under relaxed selection are expected to become reduced or disappear completely, a process called vestigialization. In parthenogenetic populations, traits historically involved in sexual reproduction are no longer under selection and potentially subject to such reduction. In Leptopilina clavipes, thelytokous (parthenogenetic) populations are infected by Wolbachia bacteria. Arrhenotokous populations do not harbor Wolbachia. When antibiotics are applied to infected females, they are cured from their infection and males arise. Such males are capable of producing offspring with uninfected females, but with lower fertilization success than sexual males. This can be attributed to the lack of selection on male fertility in thelytokous lines. In this study we used this variation in L. clavipes male fertility to determine the genetic basis of this trait. Males from cured thelytokous populations were crossed to females from uninfected populations. Using AFLP markers, a genetic linkage map was generated, consisting of five linkage groups and spanning a total distance of 219.9 cM. A single QTL of large effect (explaining 46.5% of the phenotypic variance) was identified for male fertility, which we call male fertility factor (mff). We discuss possible mechanisms underlying the effect of mff, as well as mechanisms involved in vestigialization of traits involved in sexual reproduction.}, } @article {pmid15454536, year = {2004}, author = {Mouton, L and Dedeine, F and Henri, H and Boulétreau, M and Profizi, N and Vavre, F}, title = {Virulence, multiple infections and regulation of symbiotic population in the Wolbachia-Asobara tabida symbiosis.}, journal = {Genetics}, volume = {168}, number = {1}, pages = {181-189}, pmid = {15454536}, issn = {0016-6731}, mesh = {Analysis of Variance ; Animals ; Body Weights and Measures ; DNA Primers ; Locomotion/physiology ; Oogenesis/physiology ; Population Density ; Population Dynamics ; Reproduction/physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Species Specificity ; *Symbiosis ; Video Recording ; Virulence ; Wasps/*microbiology/*physiology ; Wolbachia/*pathogenicity/physiology ; }, abstract = {The density and regulation of microbial populations are important factors in the success of symbiotic associations. High bacterial density may improve transmission to the next generation, but excessive replication could turn out to be costly to the host and result in higher virulence. Moreover, differences in virulence may also depend on the diversity of symbionts. Using the maternally transmitted symbiont Wolbachia, we investigated how bacterial density and diversity are regulated and influence virulence in host insects subject to multiple infection. The model we used was the wasp Asobara tabida that naturally harbors three different Wolbachia strains, of which two are facultative and induce cytoplasmic incompatibility, whereas the third is necessary for the host to achieve oogenesis. Using insect lines infected with different subsets of Wolbachia strains, we show that: (i) some traits of A. tabida are negatively affected by Wolbachia; (ii) the physiological cost increases with the number of co-infecting strains, which also corresponds to an increase in the total bacterial density; and (iii) the densities of the two facultative Wolbachia strains are independent of one another, whereas the obligatory strain is less abundant when it is alone, suggesting that there is some positive interaction with the other strains.}, } @article {pmid15386106, year = {2004}, author = {Rowley, SM and Raven, RJ and McGraw, EA}, title = {Wolbachia pipientis in Australian spiders.}, journal = {Current microbiology}, volume = {49}, number = {3}, pages = {208-214}, doi = {10.1007/s00284-004-4346-z}, pmid = {15386106}, issn = {0343-8651}, mesh = {Animals ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Phylogeny ; Polymerase Chain Reaction ; Queensland ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Spiders/*microbiology ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia pipientis is an endosymbiotic bacterium common to arthropods and filarial nematodes. This study presents the first survey and characterization of Wolbachia pipientis that infect spiders. All spiders were collected from Queensland, Australia during 2002-2003 and screened for Wolbachia infection using PCR approaches. The Wolbachia strains present in the spiders are diverse, paraphyletic, and for the most part closely related to strains that infect insects. We have also identified several spider Wolbachia strains that form a lineage outside the currently recognized six main Wolbachia supergroups (A-F). Incongruence between spider and Wolbachia phylogenies indicates a history of horizontal transmission of the bacterium in these host taxa. Like other arthropods, spiders are capable of harboring multiple Wolbachia strains.}, } @article {pmid15385467, year = {2004}, author = {Gillette-Ferguson, I and Hise, AG and McGarry, HF and Turner, J and Esposito, A and Sun, Y and Diaconu, E and Taylor, MJ and Pearlman, E}, title = {Wolbachia-induced neutrophil activation in a mouse model of ocular onchocerciasis (river blindness).}, journal = {Infection and immunity}, volume = {72}, number = {10}, pages = {5687-5692}, pmid = {15385467}, issn = {0019-9567}, support = {P30 EY011373/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; K08 AI054652/AI/NIAID NIH HHS/United States ; R01 EY010320/EY/NEI NIH HHS/United States ; AI07024/AI/NIAID NIH HHS/United States ; T32 AI007024/AI/NIAID NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Cell Extracts ; Chemokines/biosynthesis/immunology ; Cornea/immunology/microbiology/parasitology/pathology ; Cytokines/biosynthesis/immunology ; *Disease Models, Animal ; Filarioidea/*microbiology/physiology ; Mice ; *Neutrophil Activation ; Neutrophils/cytology/*immunology/*microbiology ; Onchocerciasis, Ocular/*immunology/microbiology/parasitology/pathology ; Phagocytosis ; Stromal Cells/immunology/microbiology/parasitology ; Vacuoles/microbiology ; Wolbachia/*immunology/isolation & purification ; }, abstract = {Endosymbiotic Wolbachia bacteria are abundant in the filarial nematodes that cause onchocerciasis (river blindness), including the larvae (microfilariae) that migrate into the cornea. Using a mouse model of ocular onchocerciasis, we recently demonstrated that it is these endosymbiotic bacteria rather than the nematodes per se that induce neutrophil infiltration to the corneal stroma and loss of corneal clarity (Saint Andre et al., Science 295:1892-1895, 2002). To better understand the role of Wolbachia organisms in the pathogenesis of this disease, we examined the fate of these bacteria in the cornea by immunoelectron microscopy. Microfilariae harboring Wolbachia organisms were injected into mouse corneas, and bacteria were detected with antibody to Wolbachia surface protein. Within 18 h of injection, neutrophils completely surrounded the nematodes and were in close proximity to Wolbachia organisms. Wolbachia surface protein labeling was also prominent in neutrophil phagosomes, indicating neutrophil ingestion of Wolbachia organisms. Furthermore, the presence of numerous electron-dense granules around the phagosomes indicated that neutrophils were activated. To determine if Wolbachia organisms directly activate neutrophils, peritoneal neutrophils were incubated with either parasite extracts containing Wolbachia organisms, parasite extracts depleted of Wolbachia organisms (by antibiotic treatment of worms), or Wolbachia organisms isolated from filarial nematodes. After 18 h of incubation, we found that isolated Wolbachia organisms stimulated production of tumor necrosis factor alpha and CXC chemokines macrophage inflammatory protein 2 and KC by neutrophils in a dose-dependent manner. Similarly, these cytokines were induced by filarial extracts containing Wolbachia organisms but not by Wolbachia-depleted extracts. Taken together, these findings indicate that neutrophil activation is an important mechanism by which Wolbachia organisms contribute to the pathogenesis of ocular onchocerciasis.}, } @article {pmid15383308, year = {2004}, author = {Fenn, K and Blaxter, M}, title = {Quantification of Wolbachia bacteria in Brugia malayi through the nematode lifecycle.}, journal = {Molecular and biochemical parasitology}, volume = {137}, number = {2}, pages = {361-364}, doi = {10.1016/j.molbiopara.2004.06.012}, pmid = {15383308}, issn = {0166-6851}, mesh = {Animals ; Base Sequence ; Brugia malayi/growth & development/*microbiology ; Colony Count, Microbial ; DNA, Bacterial/genetics ; Female ; Genome, Bacterial ; Male ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, } @article {pmid15375144, year = {2004}, author = {Gómez-Valero, L and Soriano-Navarro, M and Pérez-Brocal, V and Heddi, A and Moya, A and García-Verdugo, JM and Latorre, A}, title = {Coexistence of Wolbachia with Buchnera aphidicola and a secondary symbiont in the aphid Cinara cedri.}, journal = {Journal of bacteriology}, volume = {186}, number = {19}, pages = {6626-6633}, pmid = {15375144}, issn = {0021-9193}, mesh = {Animals ; Aphids/*microbiology ; Buchnera/genetics/*isolation & purification ; DNA, Ribosomal/chemistry ; Microscopy, Electron ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Intracellular symbiosis is very common in the insect world. For the aphid Cinara cedri, we have identified by electron microscopy three symbiotic bacteria that can be characterized by their different sizes, morphologies, and electrodensities. PCR amplification and sequencing of the 16S ribosomal DNA (rDNA) genes showed that, in addition to harboring Buchnera aphidicola, the primary endosymbiont of aphids, C. cedri harbors a secondary symbiont (S symbiont) that was previously found to be associated with aphids (PASS, or R type) and an alpha-proteobacterium that belongs to the Wolbachia genus. Using in situ hybridization with specific bacterial probes designed for symbiont 16S rDNA sequences, we have shown that Wolbachia was represented by only a few minute bacteria surrounding the S symbionts. Moreover, the observed B. aphidicola and the S symbionts had similar sizes and were housed in separate specific bacterial cells, the bacteriocytes. Interestingly, in contrast to the case for all aphids examined thus far, the S symbionts were shown to occupy a similarly sized or even larger bacteriocyte space than B. aphidicola. These findings, along with the facts that C. cedri harbors the B. aphidicola strain with the smallest bacterial genome and that the S symbionts infect all Cinara spp. analyzed so far, suggest the possibility of bacterial replacement in these species.}, } @article {pmid15354833, year = {2004}, author = {Voronin, DA and Dudkina, NV and Kiseleva, EV}, title = {A new form of symbiotic bacteria Wolbachia found in the endoplasmic reticulum of early embryos of Drosophila melanogaster.}, journal = {Doklady biological sciences : proceedings of the Academy of Sciences of the USSR, Biological sciences sections}, volume = {396}, number = {}, pages = {227-229}, pmid = {15354833}, issn = {0012-4966}, mesh = {Animals ; Drosophila melanogaster/embryology/*microbiology ; Embryo, Nonmammalian/microbiology/ultrastructure ; Endoplasmic Reticulum/*microbiology/ultrastructure ; Symbiosis ; Wolbachia/*isolation & purification/ultrastructure ; }, } @article {pmid15347518, year = {2004}, author = {Rigaud, T and Moreau, J}, title = {A cost of Wolbachia-induced sex reversal and female-biased sex ratios: decrease in female fertility after sperm depletion in a terrestrial isopod.}, journal = {Proceedings. Biological sciences}, volume = {271}, number = {1551}, pages = {1941-1946}, pmid = {15347518}, issn = {0962-8452}, mesh = {Analysis of Variance ; Animals ; Female ; Fertility/physiology ; *Hermaphroditic Organisms ; Isopoda/*microbiology/*physiology ; Male ; *Sex Determination Processes ; *Sex Ratio ; Sperm Count ; Spermatozoa/physiology ; *Wolbachia ; }, abstract = {A number of parasites are vertically transmitted to new host generations via female eggs. In such cases, host reproduction is an intimate component of parasite fitness and no cost of the infection on host reproduction is expected to evolve. A number of these parasites distort host sex ratios towards females, thereby increasing either parasite fitness or the proportion of the host that transmit the parasite. In terrestrial isopods (woodlice), Wolbachia bacteria are responsible for sex reversion and female-biased sex ratios, changing genetic males into functional neo-females. Although sex ratio distortion is a powerful means for parasites to increase in frequency in host populations, it also has potential consequences on host biology, which may, in turn, have consequences for parasite prevalence. We used the woodlouse Armadillidium vulgare to test whether the interaction between Wolbachia infection and the resulting excess of females would limit female fertility through the reduction in sperm number that they receive from males. We showed that multiple male mating induces sperm depletion, and that this sperm depletion affects fertility only in infected females. This decrease in fertility, associated with male mate choice, may limit the spread of Wolbachia infections in host populations.}, } @article {pmid15345422, year = {2004}, author = {Veneti, Z and Clark, ME and Karr, TL and Savakis, C and Bourtzis, K}, title = {Heads or tails: host-parasite interactions in the Drosophila-Wolbachia system.}, journal = {Applied and environmental microbiology}, volume = {70}, number = {9}, pages = {5366-5372}, pmid = {15345422}, issn = {0099-2240}, mesh = {Animals ; Blastoderm/microbiology ; Drosophila/embryology/*microbiology ; Embryo, Nonmammalian/microbiology ; Female ; *Host-Parasite Interactions ; Morphogenesis ; Ovary/microbiology ; Species Specificity ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia strains are endosymbiotic bacteria typically found in the reproductive tracts of arthropods. These bacteria manipulate host reproduction to ensure maternal transmission. They are usually transmitted vertically, so it has been predicted that they have evolved a mechanism to target the host's germ cells during development. Through cytological analysis we found that Wolbachia strains display various affinities for the germ line of Drosophila. Different Wolbachia strains show posterior, anterior, or cortical localization in Drosophila embryos, and this localization is congruent with the classification of the organisms based on the wsp (Wolbachia surface protein) gene sequence. This embryonic distribution pattern is established during early oogenesis and does not change until late stages of embryogenesis. The posterior and anterior localization of Wolbachia resembles that of oskar and bicoid mRNAs, respectively, which define the anterior-posterior axis in the Drosophila oocyte. By comparing the properties of a single Wolbachia strain in different host backgrounds and the properties of different Wolbachia strains in the same host background, we concluded that bacterial factors determine distribution, while bacterial density seems to be limited by the host. Possible implications concerning cytoplasmic incompatibility and evolution of strains are discussed.}, } @article {pmid15341153, year = {2004}, author = {Kumpulainen, T and Grapputo, A and Mappes, J}, title = {Parasites and sexual reproduction in psychid moths.}, journal = {Evolution; international journal of organic evolution}, volume = {58}, number = {7}, pages = {1511-1520}, doi = {10.1111/j.0014-3820.2004.tb01731.x}, pmid = {15341153}, issn = {0014-3820}, mesh = {Analysis of Variance ; Animals ; Body Weights and Measures ; Female ; Fertility/physiology ; Finland ; *Models, Biological ; Moths/*parasitology/*physiology ; Reproduction/physiology ; *Sex ; }, abstract = {Persistence of sexual reproduction among coexisting asexual competitors has been a major paradox in evolutionary biology. The number of empirical studies is still very limited, as few systems with coexisting sexual and strictly asexual lineages have been found. We studied the ecological mechanisms behind the simultaneous coexistence of a sexually and an asexually reproducing closely related species of psychid moth in Central Finland between 1999 and 2001. The two species compete for the same resources and are often infected by the same hymenopteran parasitoids. They are extremely morphologically and behaviorally similar and can be separated only by their reproductive strategy (sexual vs. asexual) or by genetic markers. We compared the life-history traits of these species in two locations where they coexist to test predictions of the cost-of-sex hypothesis. We did not find any difference in female size, number of larvae, or offspring survival between the sexuals and asexuals, indicating that sexuals are subject to cost of sex. We also used genetic markers to check and exclude the possibility of Wolbachia bacteria infection inducing parthenogenesis. None of the samples was infected by Wolbachia and, thus, it is unlikely that these bacteria could affect our results. We sampled 38 locations to study the prevalence of parasitoids and the moths' reproductive strategy. We found a strong positive correlation between prevalence of sexual reproduction and prevalence of parasitoids. In locations where parasitoids are rare asexuals exist in high densities, whereas in locations with a high parasitoid load the sexual species was dominant. Spatial distribution alone does not explain the results. We suggest that the parasite hypothesis for sex may offer an explanation for the persistence of sexual moths in this system.}, } @article {pmid15329666, year = {2004}, author = {Vavre, F and de Jong, JH and Stouthamer, R}, title = {Cytogenetic mechanism and genetic consequences of thelytoky in the wasp Trichogramma cacoeciae.}, journal = {Heredity}, volume = {93}, number = {6}, pages = {592-596}, doi = {10.1038/sj.hdy.6800565}, pmid = {15329666}, issn = {0018-067X}, mesh = {Animals ; Biological Evolution ; *Cytogenetic Analysis ; Female ; Genetic Markers ; Meiosis ; Mitosis ; Parthenogenesis/*genetics ; Wasps/*genetics ; }, abstract = {In Hymenoptera, complete parthenogenesis, that is thelytoky, is a common phenomenon where virgin females produce only daughters. Thelytoky is often induced by bacteria of the genus Wolbachia, but can also be genetically determined by the insect itself, as in the genus Trichogramma where both forms exist. In order to compare these two forms of thelytoky, chromosome behaviour analysis in young eggs and genetic analysis of microsatellite markers were carried out in the wasp Trichogramma cacoeciae, where thelytoky is genetically determined. Microscopic studies revealed that during female gamete formation meiotic cells undergo only a single equational division followed by the expulsion of a single polar body. This absence of meiotic recombination and reduction corresponds well with the high levels of heterozygosity observed in females collected from the field and a nonsegregation pattern in the offspring of heterozygous females. We therefore concluded that diploidy in T. cacoeciae is maintained through an apomictic cloning mechanism and that the incidence of thelytoky under genetic control of the wasp differs entirely from the mechanism induced by Wolbachia infection, where thelytoky is restored through gamete duplication.}, } @article {pmid15322925, year = {2004}, author = {Foster, J and Baldo, L and Blaxter, M and Henkle-Dührsen, K and Whitton, C and Slatko, B and Bandi, C}, title = {The bacterial catalase from filarial DNA preparations derives from common pseudomonad contaminants and not from Wolbachia endosymbionts.}, journal = {Parasitology research}, volume = {94}, number = {2}, pages = {141-146}, pmid = {15322925}, issn = {0932-0113}, mesh = {Animals ; Catalase/*genetics ; DNA, Bacterial/analysis/*genetics ; DNA, Helminth/analysis/*genetics ; DNA, Ribosomal/analysis ; Filarioidea/genetics/*microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Pseudomonas/*enzymology/genetics ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*enzymology/genetics ; }, abstract = {Wolbachia are obligatory endosymbionts in many species of filarial nematodes. Certain bacterial molecules induce antibody responses in mammalian hosts infected with filariae, while others activate inflammatory responses that contribute to pathology. These findings, coupled with antibiotic studies demonstrating the dependence of filarial embryogenesis on the presence of Wolbachia, have intensified research on Wolbachia-nematode interactions, and the effects of Wolbachia molecules on the mammalian immune system. By amplification and sequencing of 16S rDNA and catalase sequences, we show that filarial DNA samples prepared from nematodes collected under typical conditions are frequently contaminated with Pseudomonas DNA. Analysis of a published DNA fragment containing a catalase attributed to the Wolbachia of Onchocerca volvulus showed it to be most like Pseudomonas, both in terms of sequence similarity and genomic organization. Additionally, there was no obvious catalase in either of two available Wolbachia genome sequences. Contamination of filarial DNA with bacterial sequences other than Wolbachia can complicate studies of the role of these symbionts in filarial biology.}, } @article {pmid15311460, year = {2004}, author = {Tsai, KH and Lien, JC and Huang, CG and Wu, WJ and Chen, WJ}, title = {Molecular (sub) grouping of endosymbiont Wolbachia infection among mosquitoes of Taiwan.}, journal = {Journal of medical entomology}, volume = {41}, number = {4}, pages = {677-683}, doi = {10.1603/0022-2585-41.4.677}, pmid = {15311460}, issn = {0022-2585}, mesh = {Animals ; Anopheles/microbiology ; Base Sequence ; Culex/microbiology ; Culicidae/classification/growth & development/*microbiology ; DNA Primers ; DNA, Bacterial/genetics/isolation & purification ; Female ; Larva/microbiology ; Ovary/microbiology ; Parthenogenesis ; Taiwan ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {Wolbachia are maternally inherited bacteria that infect a wide range of arthropods as well as filarial worms. The infection usually results in reproductive distortions of the host, primarily cytoplasmic incompatibility, parthenogenesis, and feminization. This study showed that Wolbachia infection (15/29; 51.72%) was prevalent among field-caught mosquitoes in Taiwan. Three mosquito species were identified as having Wolbachia A infection, eight species as having Wolbachia B, and four other species were dually infected by both groups. Each Wolbachia isolate from different mosquitoes was further divided into a specific subgroup. However, there were still some isolates that did not belong to any known subgroup, suggesting that more subgroups remain to be identified. Investigation of tissue tropism in either Aedes albopictus (Skuse) or Armigeres subalbatus (Coquillett) revealed that Wolbachia were extensively distributed within the host, although the ovary was most susceptible to infection. This report provides preliminary features of molecular relationships among Wolbachia groups of mosquitoes from Taiwan.}, } @article {pmid15306370, year = {2004}, author = {Ballard, JW and James, AC}, title = {Differential fitness of mitochondrial DNA in perturbation cage studies correlates with global abundance and population history in Drosophila simulans.}, journal = {Proceedings. Biological sciences}, volume = {271}, number = {1544}, pages = {1197-1201}, pmid = {15306370}, issn = {0962-8452}, mesh = {Animals ; Cell Nucleus/genetics/physiology ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics/*physiology ; Gene Frequency ; *Genetic Variation ; Haplotypes/*genetics ; Linear Models ; Population Dynamics ; *Selection, Genetic ; }, abstract = {Mitochondria are often referred to as the powerhouse of the cell. However, research linking intraspecific differences in organismal fitness with genotypic mitochondrial DNA (mtDNA) variation has been hampered by the lack of variation in experimentally tractable species. This study examines whether fly lines harbouring three distinct Drosophila simulans mtDNA types (siI, -II and -III) exhibit differential fitness in laboratory perturbation cages. Comparison of the pre-perturbation and post-perturbation data shows that both the mtDNA and mitonuclear interactions have a significant and repeatable effect on the frequency of flies with specific genotypes in population cages (siII > -III > -I) and that coadapted mitonuclear interactions are greatest in the siI type. The rank order of mtDNA frequency correlates with the observed worldwide distribution of the haplogroups while mitonuclear interactions are most significant in the siI haplogroup that is likely to have been subject to repeated population bottlenecks. One possible explanation for the maintenance of the least fit siI haplogroup on Pacific islands is that it is protected from extinction by Wolbachia infection.}, } @article {pmid15306342, year = {2004}, author = {Sinkins, SP and Godfray, HC}, title = {Use of Wolbachia to drive nuclear transgenes through insect populations.}, journal = {Proceedings. Biological sciences}, volume = {271}, number = {1546}, pages = {1421-1426}, pmid = {15306342}, issn = {0962-8452}, mesh = {Animals ; Cell Nucleus/genetics ; Cytoplasm/genetics ; *Genetics, Population ; Insecta/*genetics/*microbiology ; *Models, Genetic ; Population Dynamics ; Time Factors ; Transgenes/*genetics ; Wolbachia/*genetics ; }, abstract = {Wolbachia is an inherited intracellular bacterium found in many insects of medical and economic importance. The ability of many strains to spread through populations using cytoplasmic incompatibility, involving sperm modification and rescue, provides a powerful mechanism for driving beneficial transgenes through insect populations, if such transgenes could be inserted into and expressed by Wolbachia. However, manipulating Wolbachia in this way has not yet been achieved. Here, we demonstrate theoretically an alternative mechanism whereby nuclear rather than cytoplasmic transgenes could be driven through populations, by linkage to a nuclear gene able to rescue modified sperm. The spread of a 'nuclear rescue construct' occurs as long as the Wolbachia show imperfect maternal transmission under natural conditions and/or imperfect rescue of modified sperm. The mechanism is most efficient when the target population is already infected with Wolbachia at high frequency, whether naturally or by the sequential release of Wolbachia-infected individuals and subsequently the nuclear rescue construct. The results provide a potentially powerful addition to the few insect transgene drive mechanisms that are available.}, } @article {pmid15306297, year = {2004}, author = {Hiroki, M and Tagami, Y and Miura, K and Kato, Y}, title = {Multiple infection with Wolbachia inducing different reproductive manipulations in the butterfly Eurema hecabe.}, journal = {Proceedings. Biological sciences}, volume = {271}, number = {1549}, pages = {1751-1755}, pmid = {15306297}, issn = {0962-8452}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; Butterflies/*microbiology/physiology ; Cluster Analysis ; Cytoplasm/microbiology ; Male ; Molecular Sequence Data ; *Phylogeny ; Reproduction/physiology ; Sequence Analysis, DNA ; Sex Characteristics ; Sex Ratio ; Species Specificity ; Spermatozoa/physiology ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are rickettsial intracellular symbionts of arthropods and nematodes. In arthropods, they act as selfish genetic elements and manipulate host reproduction, including sex-ratio distortion and cytoplasmic incompatibility (CI). Previous studies showed that infection of feminizing Wolbachia and CI Wolbachia sympatrically occurred in the butterfly Eurema hecabe. We demonstrate that feminization-infecting individuals can rescue sperm modified by CI-infecting males. Phylogenetic analysis revealed that feminized individuals are infected with two distinct Wolbachia strains: one is shared with CI-inducing matrilines, and the other is only found in feminized matrilines. Therefore, the simultaneous double manipulation, CI rescue and feminization, is caused by different Wolbachia strains in feminized individuals, not by a single Wolbachia with two functions. This is the first finding of double infection of Wolbachia with different reproductive manipulations.}, } @article {pmid15305681, year = {2004}, author = {Sacchi, L}, title = {[Ultrastructural basis of interactions between prokaryotes and eukaryotes in different symbiotic models].}, journal = {Parassitologia}, volume = {46}, number = {1-2}, pages = {19-24}, pmid = {15305681}, issn = {0048-2951}, mesh = {Animals ; *Bacterial Physiological Phenomena ; Bdellovibrio/physiology/ultrastructure ; Biological Evolution ; Brugia pahangi/microbiology/ultrastructure ; Cockroaches/cytology/embryology/microbiology ; Eggs/microbiology ; Eukaryotic Cells/*ultrastructure ; Fat Body/microbiology ; Female ; Filarioidea/cytology/*microbiology ; Hemocytes/microbiology ; Insecta/cytology/*microbiology ; Isoptera/cytology/microbiology ; Models, Biological ; Ovary/microbiology ; Prokaryotic Cells/*ultrastructure ; *Symbiosis ; Ticks/cytology/*microbiology ; Wolbachia/physiology/ultrastructure ; }, abstract = {This paper reviews the Author's contribution to the knowledge of the ultrastructural basis of the prokaryote-eukaryote interactions in different models assessed by an ultrastructural approach. In agreement with the hypothesis of the origin of eukaryotic cells, which are chimeras of several prokaryotes with different morpho-functional specializations, symbiosis had major consequence for evolution of life. In Arthropods, one of the most successful lifestyles, the presence of endosymbiotic prokaryotes, plays an important role in their metabolism. In some cases, genome integration has occurred in the endosymbiotic relationships with the host, proving that intracellular symbiosis is not merely a nutritional supplement. Intracellular symbiotic bacteria are also described in nematodes. In particular, the presence of intracellular Wolbachia in filariae, even if its function is not yet completely known, influences positively the reproductive biology and the survival of the host, as proved by antibiotic treatment against this bacterium. The ultrastructural images reported in this review were obtained using different species of cockroaches, termites, ticks and filarial nematodes. The traditional methods of transmission (TEM), scansion (SEM) and immuno electron microscopy were used. In addition, also freeze-fracture and deep-etching techniques were employed. The cockroaches and the primitive termite Mastotermes darwiniensis host symbiotic bacteria in the ovary and in specialized cells (bacteriocytes) of the fat body. These bacteria have the typical cell boundary profile of gram-negative bacteria and are enveloped in a vacuolar membrane produced by the host cell. Molecular sequence data of 16S rDNA of endosymbionts of five species of cockroaches and M. darwiniensis indicate that they are members of the Flavobacteria-bacteroides group and that the infection occurred in an ancestor common to cockroaches and termites probably after the end of the Paleozoic (250 Ma BP). The symbiotic bacteria are transmitted transovarially and, during embryogenesis, they are integrated into the morphogenetic processes. In particular, we were able to demonstrate that the origin of the bacteriocyte should be looked for in the cells of the haemocyte line (embryonic plasmatocytes). The eggs are infected by the bacteria emerging from the bacteriocytes of the ovaric fat body and, at the end of the vitellogenesis, they are actively phagocytized by the egg membrane. In filarial nematodes, intracellular bacteria belonging to the genus Wolbachia have been described: they have evolved an obligatory mutualistic association with their host. In fact, antibiotic treatments lead to the clearance of bacteria and this loss produces a negative impact on reproduction and survival of the filarial host. We evidenced, by TEM, the degenerative events occurring during the embriogenesis of Brugia pahangi and Dirofilaria immitis after tetracycline treatment. The data suggest that the Wolbachia play a direct role in worm metabolism. Finally, a new additional model of the prokaryote-eukaryote interaction has been described: we have recently discovered a new intracellular alpha-proteobacterium, named Iric ES1, which resides in the ovarian tissues of the tick Ixodes ricinus. The intriguing characteristic of this bacterium is its ability to invade and consume the ovaric mitochondria. From an evolutionary perspective, it is interesting to note that Iric ES1 enters mitochondria in a similar way to that employed by the "predatory" bacterium Bdellovibrio bacteriovorus.}, } @article {pmid15305172, year = {2004}, author = {Fry, AJ and Palmer, MR and Rand, DM}, title = {Variable fitness effects of Wolbachia infection in Drosophila melanogaster.}, journal = {Heredity}, volume = {93}, number = {4}, pages = {379-389}, doi = {10.1038/sj.hdy.6800514}, pmid = {15305172}, issn = {0018-067X}, mesh = {Animals ; Drosophila melanogaster/*microbiology ; Female ; Fertility/physiology ; Histocompatibility/physiology ; Male ; Polymerase Chain Reaction ; Survival ; Symbiosis/*physiology ; Tetracycline Resistance/physiology ; Wolbachia/*physiology ; }, abstract = {Maternally inherited Wolbachia bacteria are extremely widespread among insects and their presence is usually associated with parasitic modifications of host fitness. Wolbachia pipientis infects Drosophila melanogaster populations from all continents, but their persistence in this species occurs despite any strong parasitic effects. Here, we have investigated the symbiosis between Wolbachia and D. melanogaster and found that Wolbachia infection can have significant survival and fecundity effects. Relative to uninfected flies, infected females from three fly strains showed enhanced survival or fecundity associated with Wolbachia infection, one strain showed both and one strain responded positively to Wolbachia removal. We found no difference in egg hatch rates (cytoplasmic incompatibility) for crosses between infected males and uninfected females, although there were fecundity differences. Females from this cross consistently produced fewer eggs than infected females and these fecundity differences could promote the spread of infection just like cytoplasmic incompatibility. More surprising, we found that infected females often had the greatest fecundity when mated to uninfected males. This could also promote the spread of Wolbachia infection, though here the fitness benefits would also help to spread infection when Wolbachia are rare. We suggest that variable fitness effects, in both sexes, and which interact strongly with the genetic background of the host, could increase cytoplasmic drive rates in some genotypes and help explain the widespread persistence of Wolbachia bacteria in D. melanogaster populations. These interactions may further explain why many D. melanogaster populations are polymorphic for Wolbachia infection. We discuss our results in the context of host-symbiont co-evolution.}, } @article {pmid15297923, year = {2004}, author = {Ruang-areerate, T and Kittayapong, P and McGraw, EA and Baimai, V and O'Neill, SL}, title = {Wolbachia replication and host cell division in Aedes albopictus.}, journal = {Current microbiology}, volume = {49}, number = {1}, pages = {10-12}, doi = {10.1007/s00284-003-4245-8}, pmid = {15297923}, issn = {0343-8651}, mesh = {Aedes/cytology/*microbiology ; Animals ; Cell Division/physiology ; DNA Replication ; Host-Parasite Interactions/immunology ; Symbiosis/*physiology ; Virulence ; Wolbachia/growth & development/pathogenicity/*physiology ; }, abstract = {Wolbachia pipientis is an obligate intracellular endosymbiont of a range of arthropod species. The microbe is best known for its manipulations of host reproduction that include inducing cytoplasmic incompatibility, parthenogenesis, feminization, and male-killing. Like other vertically transmitted intracellular symbionts, Wolbachia's replication rate must not outpace that of its host cells if it is to remain benign. The mosquito Aedes albopictus is naturally infected both singly and doubly with different strains of Wolbachia pipientis. During diapause in mosquito eggs, no host cell division is believed to occur. Further development is triggered only by subsequent exposure of the egg to water. This study uses diapause in Wolbachia-infected Aedes albopictus eggs to determine whether symbiont replication slows or stops when host cell division ceases or whether it continues at a low but constant rate. We have shown that Wolbachia densities in eggs are greatest during embryonation and then decline throughout diapause, suggesting that Wolbachia replication is dependent on host cell replication.}, } @article {pmid15287587, year = {2004}, author = {McGarry, HF and Egerton, GL and Taylor, MJ}, title = {Population dynamics of Wolbachia bacterial endosymbionts in Brugia malayi.}, journal = {Molecular and biochemical parasitology}, volume = {135}, number = {1}, pages = {57-67}, doi = {10.1016/j.molbiopara.2004.01.006}, pmid = {15287587}, issn = {0166-6851}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Brugia malayi/genetics/isolation & purification/*microbiology ; Colony Count, Microbial ; Culicidae/parasitology ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/analysis ; DNA, Helminth/analysis ; Genes, Bacterial ; Genes, Helminth ; Gerbillinae/parasitology ; Microscopy, Electron ; Polymerase Chain Reaction ; Population Dynamics ; Symbiosis ; Wolbachia/genetics/*growth & development ; }, abstract = {The human filarial nematode Brugia malayi contains an endosymbiotic bacterium, Wolbachia. We used real-time quantitative polymerase chain reaction (QPCR) and microscopy to investigate the population dynamics of the bacterium-nematode association. Two Wolbachia (wsp and ftsZ) and one nematode (gst) genes were amplified from all life-cycle stages of B. malayi and results expressed as gene copies per worm and as Wolbachia/nematode ratios. Since the genes were single copy and there was one genome per Wolbachia, the gene copy numbers were equivalent to the numbers of bacteria. These were similar in microfilariae and the mosquito-borne larval stages (L2 and L3), with the lowest ratios of Wolbachia/nematode DNA. However, within 7 days of infection of the mammalian host, bacteria had increased 600-fold and the bacteria/worm ratio was the highest of all life-cycle stages. The rapid multiplication continued throughout L4 development, so that the major period of bacterial population growth occurred within 4 weeks of infection of the definitive host. Microscopy confirmed that there were few bacteria in mosquito-derived L3 but many, in large groups, in L4 collected 9 and 21 days after infection. In adult male worms up to 15 months of age, the bacterial populations were maintained, whilst in females, bacteria numbers increased as the worms matured and as the ovary and embryonic larval stages became infected. These results support the hypothesis that the bacteria are essential for larval development in the mammalian host and for the long-term survival of adult worms.}, } @article {pmid19712416, year = {2004}, author = {Czarnetzki, AB and Tebbe, CC}, title = {Diversity of bacteria associated with Collembola - a cultivation-independent survey based on PCR-amplified 16S rRNA genes.}, journal = {FEMS microbiology ecology}, volume = {49}, number = {2}, pages = {217-227}, doi = {10.1016/j.femsec.2004.03.007}, pmid = {19712416}, issn = {0168-6496}, abstract = {The bacterial communities found in eight different soil-inhabiting microarthropod species of the Class Collembola (springtails) were analyzed by analysis of PCR-amplified 16S rRNA genes obtained without cultivation from total DNA. To characterize the bacteria associated with the parthenogenetically-reproducing Folsomia candida, the almost complete 16S rRNA genes were amplified with three universal primers, i.e., the forward primer F27 and the reverse primer R1492 or R1525. With the reverse primer R1492, 100% of 49 cloned PCR products were identical to the 16S rRNA gene of the intracellular reproduction parasite Wolbachia pipientis (Supergroup E). However, no Wolbachia was detected with the reverse primer R1525 when two different breeding stocks of F. candida were analyzed. Clone libraries from one breeding stock were composed exclusively of a sequence related closely to intracellular bacteria of the genus Rickettsiella (Gammaproteobacteria) (93 clones). In contrast, this sequence was not detected in the other F. candida breeding stock. Instead, sequences of 95 clones originated from different phylogenetic groups (Alphaproteobacteria, Gammaproteobacteria, Firmicutes and Planctomycetes). Several were closely related to bacteria, which are known to live in soil and interact with insects. Genetic profiles based on PCR-amplified partial 16S rRNA genes and single-strand conformation polymorphism (SSCP) indicated that different Collembola species harbored different bacterial communities. SSCP bands indicating Wolbachia pipientis supergroup E were detected in profiles from four of the five parthenogenetic species included in this study. Other dominant bands in the SSCP profiles were related to bacteria from the phyla Proteobacteria, Actinobacteria, Firmicutes and Bacteroidetes. One sequence (AJ605704) indicated the presence of a not-yet-described intracellular symbiont from the Gammaproteobacteria, but several other sequences may represent less specific interactions between environmental bacteria and Collembola.}, } @article {pmid15281058, year = {2004}, author = {Vogt, G and Tolley, L and Scholtz, G}, title = {Life stages and reproductive components of the Marmorkrebs (marbled crayfish), the first parthenogenetic decapod crustacean.}, journal = {Journal of morphology}, volume = {261}, number = {3}, pages = {286-311}, doi = {10.1002/jmor.10250}, pmid = {15281058}, issn = {0362-2525}, mesh = {Animals ; Decapoda/anatomy & histology/growth & development/*physiology ; Female ; Genitalia/anatomy & histology/growth & development/ultrastructure ; Life Cycle Stages/*physiology ; Oocytes/cytology/ultrastructure ; Ovarian Follicle/cytology/ultrastructure ; Ovary/anatomy & histology/immunology/microbiology/ultrastructure ; Parthenogenesis/*physiology ; Sex Characteristics ; }, abstract = {Recently, we briefly reported on the first case of parthenogenesis in the decapod Crustacea which was found in the Marmorkrebs or marbled crayfish, a cambarid species of unknown geographic origin and species identity. Curiously, this animal is known only from aquarium populations, where it explosively propagates. By means of light and electron microscopic techniques we have now investigated the reproductive components of this crayfish, using more than 100 specimens ranging from hatchling to repeatedly spawned adult. Additionally, we documented its principal life stages. Our results revealed that the external sexual characters and also the gonads of the marbled crayfish are purely female, making this fast-reproducing species a good model for investigating female reproductive features in crayfish. Testicular tissues, ovotestes, or male gonoducts, gonopores, or gonopods were never found, either in small juveniles or large adult specimens, confirming the parthenogenetic nature of this crayfish. Parthenogenesis may have arisen spontaneously or by interspecific hybridization since Wolbachia-like feminizing microorganisms were not found in the ovaries. The external sexual characters of the marbled crayfish are first recognized in Stage 4 juveniles and are structurally complete approximately 2 months after hatching in specimens of approximately 2 cm total length. In the same life stage the ovary is fully differentiated as well, although the oocytes are in previtellogenic and primary vitellogenic stages only. The architecture of the mature ovary and also the synchronous maturation of cohorts of primary vitellogenic oocytes by secondary vitellogenesis are in general agreement with data published on ovaries of bisexual crayfish. New results were obtained with respect to the muscular nature of the ovarian envelope and its extensive proliferation after the first spawning, the distribution of hemal sinuses in the ovarian envelope and in the interstitium around the oogenetic pouches, the high transport activity of the follicle cells, and the colonization of oogenetic pouches by previtellogenic oocytes that originate in the germaria. Investigation of the nuclei of oocytes in the germaria and oogenetic pouches revealed no signs of meiosis, as usually found in females of bisexual decapods, suggesting that parthenogenesis in the marbled crayfish might be an apomictic thelytoky. The detection of new rickettsial and coccidian infections in the ovary and further organs raises fears that the marbled crayfish might endanger native European species by transmission of pathogens once escaped into the wild.}, } @article {pmid15271208, year = {2004}, author = {Sanogo, YO and Dobson, SL}, title = {Molecular discrimination of Wolbachia in the Culex pipiens complex: evidence for variable bacteriophage hyperparasitism.}, journal = {Insect molecular biology}, volume = {13}, number = {4}, pages = {365-369}, doi = {10.1111/j.0962-1075.2004.00498.x}, pmid = {15271208}, issn = {0962-1075}, support = {NIH-AI-51533/AI/NIAID NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacteriophages/*genetics ; Base Sequence ; Culex/classification/genetics/*microbiology ; Cytoplasm/genetics ; DNA Primers ; Molecular Sequence Data ; *Phylogeny ; Polymorphism, Single-Stranded Conformational ; Sequence Alignment ; Sequence Analysis, DNA ; Species Specificity ; Symbiosis/*genetics ; Wolbachia/*genetics/*virology ; }, abstract = {The medically important members of the Culex pipiens species complex provide an enigma for systematists, evolutionary biologists, and vector biologists. The species complex is composed of forms that differ in their ecology, behaviour, physiology and vector competence. Cytoplasmic incompatibility (CI) caused by endosymbiotic Wolbachia bacteria is thought to play an important role in restricting gene flow and the evolution of the Culex complex. Here we describe the first molecular marker useful for discriminating between Wolbachia infections in Culex. A putative bacteriophage locus (orf7) varies between Culex forms in copy number and sequence. We provide evidence that the orf7 loci are strictly associated with Wolbachia and are maternally inherited.}, } @article {pmid15271207, year = {2004}, author = {Tagami, Y and Miura, K}, title = {Distribution and prevalence of Wolbachia in Japanese populations of Lepidoptera.}, journal = {Insect molecular biology}, volume = {13}, number = {4}, pages = {359-364}, doi = {10.1111/j.0962-1075.2004.00492.x}, pmid = {15271207}, issn = {0962-1075}, mesh = {Animals ; Base Sequence ; Cluster Analysis ; DNA Primers ; Electrophoresis, Agar Gel ; Genitalia/*microbiology ; Japan ; Lepidoptera/*microbiology ; Molecular Sequence Data ; *Phylogeny ; Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; Species Specificity ; Wolbachia/*genetics ; }, abstract = {Wolbachia are cytoplasmically inherited bacteria that are reported to infect at least 18-30% of all insect species. Our survey of Lepidoptera indicated that 44.9% of forty-nine species and 77.8% of nine families tested positive for Wolbachia using PCR with wsp primers. Nineteen species had not been described previously as infected. In particular, although Pieris rapae, which is a common species in Japan, is infected by Wolbachia, the prevalence was very low (3.4%) and there were some localities where Wolbachia could not be detected. The probability of detection of Wolbachia depends on the number of screened individuals of P. rapae. The results indicate that the actual number of species that are positive for Wolbachia may be higher than previously reported.}, } @article {pmid15270538, year = {2003}, author = {Casiraghi, M and Werren, JH and Bazzocchi, C and Biserni, A and Bandi, C}, title = {dnaA gene sequences from Wolbachia pipientis support subdivision into supergroups and provide no evidence for recombination in the lineages infecting nematodes.}, journal = {Parassitologia}, volume = {45}, number = {1}, pages = {13-18}, pmid = {15270538}, issn = {0048-2951}, mesh = {Animals ; Bacterial Proteins/*genetics ; DNA, Bacterial/genetics ; DNA-Binding Proteins/*genetics ; Filarioidea/*microbiology ; Insecta/microbiology ; Phylogeny ; Recombination, Genetic ; Sequence Alignment ; Sequence Homology, Nucleic Acid ; Species Specificity ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia pipientis is an intracellular bacterial endosymbiont of arthropods and filarial nematodes. Six main supergroups of W. pipientis have been described: supergroups A, B, E, and F encompass arthropod wolbachiae; supergroups C and D encompass nematode wolbachiae. The description of these six supergroups has been based on the analysis of only two genes (ftsZ and 16S rDNA) and before decisions are taken on the taxonomic status of the six supergroups, analysis of further genes is required. In addition, the branching order of the six supergroups is still unresolved. Sequence information from other genes is also needed to allow phylogenesis to be addressed through the analysis of a higher number of characters. Here we report sequences from a portion of the gene coding for the DNAA protein of W pipientis, generated from the endosymbionts of 22 host species. Phylogenies based on dnaA gene sequences are congruent with the existence of at least six supergroups of W pipientis. In addition, subtrees generated for nematode wolbachiae in supergroups C and D were compared to the trees based on the already available gene sequences (ftsZ, 16S rDNA and wsp). The congruence observed among the trees based on the different genes agrees with the hypothesis that recombination does not occur in nematode wolbachiae.}, } @article {pmid15267003, year = {2003}, author = {Sacchi, L and Corona, S and Kramer, L and Calvi, L and Casiraghi, M and Franceschi, A}, title = {Ultrastructural evidence of the degenerative events occurring during embryogenesis of the filarial nematode Brugia pahangi after tetracycline treatment.}, journal = {Parassitologia}, volume = {45}, number = {2}, pages = {89-96}, pmid = {15267003}, issn = {0048-2951}, mesh = {Animals ; Brugia pahangi/*embryology/microbiology/ultrastructure ; Embryo, Nonmammalian/drug effects/microbiology/ultrastructure ; Female ; Male ; Microscopy, Electron ; Oocytes/drug effects/microbiology ; Ovary/cytology/drug effects ; Symbiosis/drug effects ; Tetracycline/*pharmacology ; Vacuoles/ultrastructure ; Wolbachia/*drug effects/isolation & purification ; }, abstract = {Intracellular bacteria belonging to the genus Wolbachia have been described in filarial nematodes and these microorganisms appear to have evolved an obligatory mutualistic association with their filarial hosts. In fact, antibiotic treatment leads to the clearance of bacteria from worms resulting in a block in embryogenesis and, eventually, death of adult filariae. Currently, the antifilarial action of antibiotic treatment is interpreted as a secondary consequence of the bacteriostatic activity against Wolbachia endosymbionts. Here, we demonstrate by transmission electron microscopy the degenerative events occurring during embryogenesis of Brugia pahangi after tetracycline treatment. After 56 days of treatment the cytoplasm of hypodermal cords was totally void of Wolbachia and numerous vacuoles, residual of cytolitic activity, were observed. In the ovary, the morphology of the oocytes was well conserved 33 days after treatment, but the texture of symbiotic bacteria appeared altered. After 56 days of treatment embryogenesis was dramatically affected and the terminal portion of the ovary appeared totally empty. The authors suggest that the symbiotic bacteria play a direct role in worm metabolism and a long-term bacteriostatic effect may block bacterial activity involved in the active control of cytolysis. As a consequence, the bacteriophorous vacuole is transformed into a digestive vacuole and the whole symbiotic population is disrupted.}, } @article {pmid15261775, year = {2004}, author = {Jeyaprakash, A and Hoy, MA}, title = {Multiple displacement amplification in combination with high-fidelity PCR improves detection of bacteria from single females or eggs of Metaseiulus occidentalis (Nesbitt) (Acari: Phytoseiidae).}, journal = {Journal of invertebrate pathology}, volume = {86}, number = {3}, pages = {111-116}, doi = {10.1016/j.jip.2004.05.005}, pmid = {15261775}, issn = {0022-2011}, mesh = {Animals ; Cytophaga/*genetics ; DNA, Bacterial/*isolation & purification ; Eggs/microbiology ; Female ; Mites/*microbiology ; Nucleic Acid Amplification Techniques/*methods ; Plasmids/*genetics ; Polymerase Chain Reaction/methods ; Rickettsiaceae/*genetics ; Sensitivity and Specificity ; }, abstract = {Amplifying microbial DNA by the polymerase chain reaction (PCR) from single phytoseiid mites has been difficult, perhaps due to the low titer of bacteria and to interference by the relatively larger amounts of mite genomic DNA. In this paper we evaluate the efficiency of standard and high-fidelity PCR protocols subsequent to amplification of the whole genome by a multiple displacement amplification (MDA) procedure developed by Dean et al. DNA from the phytoseiid Phytoseiulus persimilis (Athias-Henriot) was tested because it lacks a Cytophaga-like organism (CLO) and we could add known amounts of a plasmid containing a cloned 16S rRNA gene fragment from a CLO from Metaseiulus occidentalis (Nesbitt). P. persimilis genomic DNA was mixed with the serially diluted plasmid and amplified using MDA followed by either standard or high-fidelity PCR. MDA followed by high-fidelity PCR was most efficient and successfully amplified an expected 1.5-kb band from as little as 0.01fg of the plasmid, which is equivalent to about 1 copy. MDA followed by high-fidelity PCR also consistently amplified Wolbachia- or CLO-specific products from naturally infected single females or eggs of M. occidentalis, which will allow detailed studies of infection frequency and transmission of several microorganisms associated with this predatory mite.}, } @article {pmid15254259, year = {2004}, author = {Bordenstein, SR and Wernegreen, JJ}, title = {Bacteriophage flux in endosymbionts (Wolbachia): infection frequency, lateral transfer, and recombination rates.}, journal = {Molecular biology and evolution}, volume = {21}, number = {10}, pages = {1981-1991}, doi = {10.1093/molbev/msh211}, pmid = {15254259}, issn = {0737-4038}, support = {R01 GM62626-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Bacteriophages/genetics/*physiology ; Base Sequence ; Evolution, Molecular ; Molecular Sequence Data ; Phylogeny ; *Recombination, Genetic ; *Transduction, Genetic ; Wolbachia/*virology ; }, abstract = {The highly specialized genomes of bacterial endosymbionts typically lack one of the major contributors of genomic flux in the free-living microbial world-bacteriophages. This study yields three results that show bacteriophages have, to the contrary, been influential in the genome evolution of the most prevalent bacterial endosymbiont of invertebrates, Wolbachia. First, we show that bacteriophage WO is more widespread in Wolbachia than previously recognized, occurring in at least 89% (35/39) of the sampled genomes. Second, we show through several phylogenetic approaches that bacteriophage WO underwent recent lateral transfers between Wolbachia bacteria that coinfect host cells in the dipteran Drosophila simulans and the hymenopteran Nasonia vitripennis. These two cases, along with a previous report in the lepidopteran Ephestia cautella, support a general mechanism for genetic exchange in endosymbionts--the "intracellular arena" hypothesis--in which genetic material moves horizontally between bacteria that coinfect the same intracellular environment. Third, we show recombination in this bacteriophage; in the region encoding a putative capsid protein, the recombination rate is faster than that of any known recombining genes in the endosymbiont genome. The combination of these three lines of genetic evidence indicates that this bacteriophage is a widespread source of genomic instability in the intracellular bacterium Wolbachia and potentially the invertebrate host. More generally, it is the first bacteriophage implicated in frequent lateral transfer between the genomes of bacterial endosymbionts. Gene transfer by bacteriophages could drive significant evolutionary change in the genomes of intracellular bacteria that are typically considered highly stable and prone to genomic degradation.}, } @article {pmid15252981, year = {2004}, author = {Weeks, AR and Stouthamer, R}, title = {Increased fecundity associated with infection by a cytophaga-like intracellular bacterium in the predatory mite, Metaseiulus occidentalis.}, journal = {Proceedings. Biological sciences}, volume = {271 Suppl 4}, number = {Suppl 4}, pages = {S193-5}, pmid = {15252981}, issn = {0962-8452}, mesh = {Analysis of Variance ; Animals ; California ; Crosses, Genetic ; *Cytophaga ; Female ; Fertility/physiology ; Host-Parasite Interactions ; Male ; Mites/*microbiology/physiology ; Ovum/physiology ; Sex Ratio ; }, abstract = {The endosymbiont Wolbachia has gained widespread notoriety over the past decade because of its high infection frequency among arthropods, and the unique heterogeneity of the host reproductive effects that it has been implicated as causing to enhance its own spread. Recently, another endosymbiotic bacterium from the Cytophaga-Flavobacterium-Bacteroides phylum has been shown to be widespread among arthropods and manipulate its hosts' reproduction to enhance its own spread. We show that infection by this Cytophaga-like organism (CLO) in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae) is associated with a significant increase in the fecundity of infected females. This adds to the growing list of phenotypes that the CLO can induce in its hosts, which now include feminization, parthenogenesis induction, cytoplasmic incompatibility and fecundity enhancement, rivalling Wolbachia for overall diversity of host reproductive manipulations.}, } @article {pmid15245413, year = {2004}, author = {Keller, GP and Windsor, DM and Saucedo, JM and Werren, JH}, title = {Reproductive effects and geographical distributions of two Wolbachia strains infecting the Neotropical beetle, Chelymorpha alternans Boh. (Chrysomelidae, Cassidinae).}, journal = {Molecular ecology}, volume = {13}, number = {8}, pages = {2405-2420}, doi = {10.1111/j.1365-294X.2004.02213.x}, pmid = {15245413}, issn = {0962-1083}, mesh = {Animals ; Base Sequence ; Coleoptera/genetics/*microbiology/physiology ; Crosses, Genetic ; DNA Primers ; Electron Transport Complex IV/genetics ; *Environment ; *Genetic Variation ; Geography ; Haplotypes/genetics ; Molecular Sequence Data ; Panama ; Population Dynamics ; Reproduction/physiology ; Sequence Analysis, DNA ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally inherited endocellular bacteria known to alter insect host reproduction to facilitate their own transmission. Multiple Wolbachia infections are more common in tropical than temperate insects but few studies have investigated their dynamics in field populations. The beetle, Chelymorpha alternans, found throughout the Isthmus of Panama, is infected with two strains of Wolbachia, wCalt1 (99.2% of beetles) and wCalt2 (53%). Populations infected solely by the wCalt1 strain were limited to western Pacific Panama, whereas populations outside this region were either polymorphic for single (wCalt1) and double infections (wCalt1 + wCalt2) or consisted entirely of double infections. The wCalt2 strain was not found as a single infection in the wild. Both strains caused cytoplasmic incompatibility (CI). The wCalt1 strain caused weak CI (approximately 20%) and the double infection induced moderate CI (approximately 70-90%) in crosses with uninfected beetles. The wCalt1 strain rescued about 75% of eggs fertilized by sperm from wCalt2 males. Based on the relationships of beetle mtDNA and infection status, maternal transmission, and repeated population sampling we determined that the double infection invaded C. alternans populations about 100,000 years ago and that the wCalt2 strain appears to be declining in some populations, possibly due to environmental factors. This may be the first study to demonstrate an association between widespread strain loss and environmental factors in the field.}, } @article {pmid15242750, year = {2004}, author = {Azzouna, A and Greve, P and Martin, G}, title = {Sexual differentiation traits in functional males with female genital apertures (male symbol fga) in the woodlice Armadillidium vulgare Latr. (Isopoda, Crustacea).}, journal = {General and comparative endocrinology}, volume = {138}, number = {1}, pages = {42-49}, doi = {10.1016/j.ygcen.2004.05.003}, pmid = {15242750}, issn = {0016-6480}, mesh = {Animals ; Cell Nucleus/ultrastructure ; Crustacea/*physiology ; DNA/biosynthesis/genetics ; Endoplasmic Reticulum/ultrastructure ; Female ; Genitalia/*anatomy & histology/cytology/*physiology ; Gonads/anatomy & histology/cytology/physiology ; Male ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Sex Differentiation/*physiology ; }, abstract = {This study reports the results of examination of the gonadal morphology and ultrastructural features of the androgenic hormone (AH)-producing androgenic gland cells of laboratory stocks of functional male woodlice, Armadillidium vulgare, with female genital apertures (male symbol fga), with and without experimentally induced infections of the sex-ratio-distorting endobacterial parasite, Wolbachia. Males (male symbol fga) have been reported in wild populations containing individuals infected with this maternally transmitted sex-ratio-distorting parasite. We report a reduction of testicular segment (utricle) number, androgenic gland cell hypertrophy, and electron-dense ultrastructural cytological features in male symbol fga males. The presence of the Wolbachia parasite had no effect on the features we examined. These results suggest that male symbol fga males are produced as the result of a delayed expression/action of the male sex-determining AH which causes a "lag-phase" delay in male differentiation in genetic males and is not due to the presence, in genetic females, of a hypothetical, epigenetic "M" gene as suggested by Rigaud and Juchault. Our results favor the interpretation of males as true genetic (ZZ) males in which the delayed AH action appears to involve cellular AH trafficking pathways which may be controlled by an impaired autosomal gene responsible for AH action.}, } @article {pmid15242714, year = {2004}, author = {Sinkins, SP}, title = {Wolbachia and cytoplasmic incompatibility in mosquitoes.}, journal = {Insect biochemistry and molecular biology}, volume = {34}, number = {7}, pages = {723-729}, doi = {10.1016/j.ibmb.2004.03.025}, pmid = {15242714}, issn = {0965-1748}, mesh = {Aedes/*microbiology ; Animals ; Culicidae/immunology/*microbiology ; Drosophila ; Female ; Host-Parasite Interactions ; Polymerase Chain Reaction ; Population Dynamics ; Wolbachia/genetics/immunology/*pathogenicity ; }, abstract = {Wolbachia are maternally inherited bacteria that induce cytoplasmic incompatibility in mosquitoes, and are able to use these patterns of sterility to spread themselves through populations. For this reason they have been proposed as a gene drive system for mosquito genetic replacement, as well as for the reduction of population size or for modulating population age structure in order to reduce disease transmission. Here, recent progress in the study of mosquito Wolbachia is reviewed. We now have much more comprehensive estimates of the parameters that can affect the spread of Wolbachia through natural populations from low starting frequencies, and for waves of spread to be maintained in the face of partial barriers to gene flow. In Aedes albopictus these dynamics are extremely favourable, with very high maternal transmission fidelity and levels of incompatibility recorded. Correspondence between measurements taken in the lab and field is much better than in the Drosophila simulans model system. Important research goals are also discussed, including Wolbachia transformation, interspecific transfer and the elucidation of the mechanisms of incompatibility and rescue; all will be aided by a wealth of new Wolbachia genome information.}, } @article {pmid15242712, year = {2004}, author = {Rasgon, JL and Scott, TW}, title = {Impact of population age structure on Wolbachia transgene driver efficacy: ecologically complex factors and release of genetically modified mosquitoes.}, journal = {Insect biochemistry and molecular biology}, volume = {34}, number = {7}, pages = {707-713}, doi = {10.1016/j.ibmb.2004.03.023}, pmid = {15242712}, issn = {0965-1748}, support = {GM-20092/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*genetics/growth & development ; Aging ; Animals ; Animals, Genetically Modified/growth & development/*microbiology ; Anopheles/*genetics/growth & development ; Culex/genetics/growth & development ; Culicidae/*genetics/growth & development ; Models, Biological ; Population ; Wolbachia/*genetics/growth & development ; }, abstract = {Wolbachia symbionts hold theoretical promise as a way to drive transgenes into insect vector populations for disease prevention. For simplicity, current models of Wolbachia dynamics and spread ignore ecologically complex factors such as the age structure of vector populations and overlapping vector generations. We developed a model including these factors to assess their impact on the process of Wolbachia spread into populations of three mosquito species (Anopheles gambiae, Aedes aegypti and Culex pipiens). Depending on the mosquito species, Wolbachia parameters, released mosquito life stage and initial age structure of the target population, the number of Wolbachia-infected mosquitoes that we predict would need to be released ranged from less than the threshold calculated by the simple model to a 10-30-fold increase. Transgenic releases into age-structured populations, which is an expectation for wild mosquitoes, will be difficult and depending on the circumstances may not be economically or logistically feasible due to the large number of infected mosquitoes that must be released. Our results support the perspective that understanding ecological factors is critical for designing transgenic vector-borne disease control strategies.}, } @article {pmid15241444, year = {2004}, author = {Christie, JS and Castro, JA and Oliver, P and Picornell, A and Ramon, MM and Moya, A}, title = {Fitness and life-history traits of the two major mitochondrial DNA haplotypes of Drosophila subobscura.}, journal = {Heredity}, volume = {93}, number = {4}, pages = {371-378}, doi = {10.1038/sj.hdy.6800513}, pmid = {15241444}, issn = {0018-067X}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics ; *Haplotypes ; Larva/growth & development ; Longevity/physiology ; Polymerase Chain Reaction ; *Quantitative Trait, Heritable ; Restriction Mapping ; Selection, Genetic ; Tissue Survival/physiology ; Wolbachia/pathogenicity ; }, abstract = {Mitochondrial DNA restriction site analyses on natural populations of Drosophila subobscura have proved the existence of two common, coexisting haplotypes (I and II), as well as a set of less frequent ones derived from them. To explain this distribution, experiments to date point practically to all possible genetic mechanisms being involved in the changes of gene frequencies (cytonuclear coadaptation, direct natural selection on mtDNA and genetic drift). In an attempt to find differences that help to understand the dynamics of these haplotypes and to detect the effect of selection, we measured certain fitness components and life-history traits (egg-larva and larva-adult viabilities and developmental times, longevity, resistance to desiccation and optimal density) of the two main haplotypes I and II when maintained in laboratory population cages. As a general trend, haplotype II showed a higher net fitness than haplotype I, which explains the superiority of haplotype II over haplotype I in experimental populations but not their coexistence in nature, where additional factors must be considered.}, } @article {pmid15238684, year = {2004}, author = {Rajan, TV}, title = {Relationship of anti-microbial activity of tetracyclines to their ability to block the L3 to L4 molt of the human filarial parasite Brugia malayi.}, journal = {The American journal of tropical medicine and hygiene}, volume = {71}, number = {1}, pages = {24-28}, pmid = {15238684}, issn = {0002-9637}, support = {AI 039075/AI/NIAID NIH HHS/United States ; AI 042362/AI/NIAID NIH HHS/United States ; AI 050228/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Brugia malayi/*drug effects/growth & development/microbiology ; DNA, Bacterial/chemistry/isolation & purification ; Humans ; Larva/drug effects/microbiology/physiology ; Molting/*drug effects ; Symbiosis ; Tetracycline/*pharmacology ; Wolbachia/drug effects/genetics/physiology ; }, abstract = {The nematode parasites Wuchereria bancrofti, Brugia malayi, and B. timori cause a human disease known as lymphatic filariasis, which afflicts approximately 120 million people worldwide. These organisms are known to contain endosymbiotic bacteria (Wolbachia) that are related to rickettsiae. It has been previously reported that tetracycline blocks the L3 to L4 molt of the filarial parasite B. malayi, and suggested that this was related to their known anti-rickettsial activity. However, this interpretation was tempered by several observations. First, Wolbachia DNA could still be detected in nematodes from tetracycline-treated cultures. In addition, chloramphenicol, which has anti-rickettsial and anti-chlamydial activity, failed to inhibit the molt. These observations could not rule out the possibility that the anti-molting activity of tetracycline is due to pharmacologic activities unrelated to its anti-rickettsial functions. This study shows that chemically modified tetracycline, which does not to have anti-microbial activity, also blocks molting.}, } @article {pmid15238531, year = {2004}, author = {Zabalou, S and Charlat, S and Nirgianaki, A and Lachaise, D and Merçot, H and Bourtzis, K}, title = {Natural Wolbachia infections in the Drosophila yakuba species complex do not induce cytoplasmic incompatibility but fully rescue the wRi modification.}, journal = {Genetics}, volume = {167}, number = {2}, pages = {827-834}, pmid = {15238531}, issn = {0016-6731}, mesh = {Analysis of Variance ; Animals ; Cytoplasm/microbiology/physiology ; DNA/genetics/isolation & purification ; Drosophila/classification/genetics/*microbiology ; Gram-Negative Bacterial Infections ; Polymerase Chain Reaction ; Species Specificity ; Wolbachia/*pathogenicity ; }, abstract = {In this study, we report data about the presence of Wolbachia in Drosophila yakuba, D. teissieri, and D. santomea. Wolbachia strains were characterized using their wsp gene sequence and cytoplasmic incompatibility assays. All three species were found infected with Wolbachia bacteria closely related to the wAu strain, found so far in D. simulans natural populations, and were unable to induce cytoplasmic incompatibility. We injected wRi, a CI-inducing strain naturally infecting D. simulans, into the three species and the established transinfected lines exhibited high levels of CI, suggesting that absence of CI expression is a property of the Wolbachia strain naturally present or that CI is specifically repressed by the host. We also tested the relationship between the natural infection and wRi and found that it fully rescues the wRi modification. This result was unexpected, considering the significant evolutionary divergence between the two Wolbachia strains.}, } @article {pmid15218907, year = {2000}, author = {Jamnongluk, W and Kittayapong, P and Baisley, KJ and O'Neill, SL}, title = {Wolbachia infection and expression of cytoplasmic incompatibility in Armigeres subalbatus (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {37}, number = {1}, pages = {53-57}, doi = {10.1603/0022-2585-37.1.53}, pmid = {15218907}, issn = {0022-2585}, support = {G0700837/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Animals, Laboratory/genetics/immunology/microbiology ; Base Sequence ; Culicidae/genetics/*immunology/*microbiology ; Cytoplasm/immunology/microbiology ; DNA Primers ; Female ; Male ; Wolbachia/*growth & development/isolation & purification ; }, abstract = {Polymerase chain reaction screening revealed that Armigeres subalbatus (Coquillett), a vector of filariasis, was infected with the intracellular bacteria Wolbachia. Laboratory crosses between infected males and uninfected females resulted in less than half the number of offspring than control crosses between uninfected individuals when young (2- to 3-d-old) males were used in the cross. However, imcompatibility was lost when old (14- to 17-d-old) males were used. Field-collected females did not show detectable cytoplasmic incompatibility, and this may be because of the age at which males mate in the field. We used head pigment fluorescence levels to age field males collected from mating swarms, and found that 25-63% of swarming males were older than 13 d. Male age may be one factor influencing the observed low levels of cytoplasmic incompatibility detected in the field.}, } @article {pmid15214166, year = {2004}, author = {Dudkina, NV and Voronin, DA and Kiseleva, EV}, title = {[Structural organization and distribution of symbiotic bacteria Wolbachia in early embryos and ovaries of Drosophila melanogaster and D. simulans].}, journal = {Tsitologiia}, volume = {46}, number = {3}, pages = {208-220}, pmid = {15214166}, issn = {0041-3771}, mesh = {Animals ; Colony Count, Microbial ; Cytoplasm/microbiology ; Drosophila/embryology/*microbiology/physiology ; Drosophila melanogaster/embryology/microbiology/physiology ; Embryo, Nonmammalian/microbiology ; Female ; Microtubules/microbiology ; Oocytes/growth & development/microbiology ; Ovary/cytology/microbiology ; Spores, Bacterial ; Symbiosis ; Wolbachia/*isolation & purification/physiology ; }, abstract = {Electron microscopic and morphometric analyses of Wolbachia distribution in early embryos of Drosophila flies have demonstrated that the number of bacteria in the embryo remains constant from fertilization to blastoderm, and that afterwards the symbionts could be observed only in the polar cells. Each bacterium has a three-layer envelope, makes contacts with microtubules and moves through the cytoplasm following the actively dividing nuclei. It has been found for the first time that Wolbachia could produce secretory vacuoles in the cytoplasm of early embryos. The relative volume of Wolbachia was five times as much in the embryos of Drosophila simulans as in those of D. melanogaster (Canton S), while the survival rate of D. simulans was half as much as that of D. melanogaster. It was shown that Wolbachia could form spore-like structures in D. simulans embryos. Ultrastructural investigations of Drosophila ovaries suggest that the bacteria may be present in all ovariol cells, including the oocyte, within whose cytoplasm they are delivered to the host. The highest number of symbionts was observed in germarium cells. In ovariol cells, the bacteria gradually decrease in number as oogenesis progresses. It has been determined for the first time that the symbionts are located closely to membranes of rough endoplasmatic reticulum in follicular and nurse cells of D. melanogaster. The data obtained suggest that Wolbachia may be involved in the regulation of oocyte maturation.}, } @article {pmid15210803, year = {2004}, author = {Brattig, NW and Bazzocchi, C and Kirschning, CJ and Reiling, N and Büttner, DW and Ceciliani, F and Geisinger, F and Hochrein, H and Ernst, M and Wagner, H and Bandi, C and Hoerauf, A}, title = {The major surface protein of Wolbachia endosymbionts in filarial nematodes elicits immune responses through TLR2 and TLR4.}, journal = {Journal of immunology (Baltimore, Md. : 1950)}, volume = {173}, number = {1}, pages = {437-445}, doi = {10.4049/jimmunol.173.1.437}, pmid = {15210803}, issn = {0022-1767}, mesh = {Adolescent ; Adult ; Animals ; Bacterial Proteins/*immunology ; Cytokines/biosynthesis ; Female ; Humans ; Lymphokines/biosynthesis ; Male ; Membrane Glycoproteins/*physiology ; Membrane Proteins/immunology ; Middle Aged ; Onchocerca volvulus/*microbiology ; Onchocerciasis/*immunology ; Receptors, Cell Surface/*physiology ; Symbiosis ; Toll-Like Receptor 2 ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Wolbachia/*immunology ; }, abstract = {More than 150 million humans in tropical countries are infected by filarial nematodes which harbor intracellular bacterial endosymbionts of the genus Wolbachia (Rickettsiales). These bacteria have been implicated in adverse effects of drug treatment in filariasis. The present study provides evidence that purified major Wolbachia surface protein (rWSP) acts as an inducer of the innate immune system through TLR2 and TLR4: 1) recombinant, stringently purified rWSP elicited the release of TNF-alpha, IL-12, and IL-8 from cultured blood cells of both Onchocerca volvulus-infected and uninfected people; 2) the inflammatory response to rWSP challenge was TLR2- and TLR4-dependent as demonstrated with TLR-transfected fibroblastoid cells, as well as macrophages and dendritic cells from functional TLR-deficient mice; 3) blood cells of onchocerciasis patients exposed to rWSP also generated down-regulating mediators IL-10 and PGE(2) after 6 days of culture; 4) furthermore, rWSP-reactive IgG1 Abs were present in sera of O. volvulus-infected people but not in those of uninfected Europeans. The lack of rWSP-reactive IgE and IgG4 in serum indicated a bias toward a Th1-type adaptive immune response. Abs against rWSP stained endobacteria in living and degenerating adult O. volvulus filariae, tissue microfilariae and host tissue macrophages that apparently had engulfed microfilariae. Thus, filarial helminths, through products of their endobacteria such as WSP, acquire characteristics of a typical microbial pathogen inducing immune responses via TLR2 and TLR4.}, } @article {pmid15190188, year = {2004}, author = {Driver, C and Georgiou, A and Georgiou, G}, title = {The contribution by mitochondrially induced oxidative damage to aging in Drosophila melanogaster.}, journal = {Biogerontology}, volume = {5}, number = {3}, pages = {185-192}, doi = {10.1023/B:BGEN.0000031156.75376.e3}, pmid = {15190188}, issn = {1389-5729}, mesh = {Animals ; Circadian Rhythm ; Drosophila melanogaster/*physiology ; Lipid Peroxides/metabolism ; Mitochondria/*physiology ; *Oxidative Stress ; }, abstract = {Strains of Drosophila melanogaster have been generated that vary in the mitochondrial genome and have a common nuclear genome. Tetracycline was used to cure the strains of the endocommensal bacteria, Wolbachia, which is transmitted maternally. Variation in the tetracycline responsive agent contributed substantially to variation in longevity and production of metabolic oxidants. Changes in diurnal cycle of activity have been investigated: such changes should be responsive to metabolism-related damage because of the role of the very active neurons, retinal cells and muscle. The metabolic oxidant model predicts that longevity and production of metabolic oxidants should be negatively correlated, and that deterioration of behavior with age and metabolic oxidant production should be positively correlated. Only the first prediction was verified: rate of change in behavior correlated with metabolic oxidants, consistent with a role of metabolic oxidants in this age-related change. However, the data indicates a role for mitochondria in determination longevity, other than by production of metabolic oxidants.}, } @article {pmid15189221, year = {2004}, author = {Zchori-Fein, E and Perlman, SJ}, title = {Distribution of the bacterial symbiont Cardinium in arthropods.}, journal = {Molecular ecology}, volume = {13}, number = {7}, pages = {2009-2016}, doi = {10.1111/j.1365-294X.2004.02203.x}, pmid = {15189221}, issn = {0962-1083}, mesh = {Animals ; Arthropods/*microbiology/physiology ; Bacteroidetes/*genetics ; Base Sequence ; Cluster Analysis ; Cytoplasm/microbiology ; DNA Primers ; DNA, Ribosomal/genetics ; Models, Genetic ; Molecular Sequence Data ; *Phylogeny ; Reproduction/physiology ; Sequence Analysis, DNA ; Species Specificity ; *Symbiosis ; Wolbachia/genetics ; }, abstract = {Abstract 'Candidatus Cardinium', a recently described bacterium from the Bacteroidetes group, is involved in diverse reproduction alterations of its arthropod hosts, including cytoplasmic incompatibility, parthenogenesis and feminization. To estimate the incidence rate of Cardinium and explore the limits of its host range, 99 insect and mite species were screened, using primers designed to amplify a portion of Cardinium 16S ribosomal DNA (rDNA). These arthropods were also screened for the presence of the better-known reproductive manipulator, Wolbachia. Six per cent of the species screened tested positive for Cardinium, compared with 24% positive for Wolbachia. Of the 85 insects screened, Cardinium was found in four parasitic wasp species and one armoured scale insect. Of the 14 mite species examined, one predatory mite was found to carry the symbiont. A phylogenetic analysis of all known Cardinium 16S rDNA sequences shows that distantly related arthropods can harbour closely related symbionts, a pattern typical of horizontal transmission. However, closely related Cardinium were found to cluster among closely related hosts, suggesting host specialization and horizontal transmission among closely related hosts. Finally, the primers used revealed the presence of a second lineage of Bacteroidetes symbionts, not related to Cardinium, in two insect species. This second symbiont lineage is closely allied with other arthropod symbionts, such as Blattabacterium, the primary symbionts of cockroaches, and male-killing symbionts of ladybird beetles. The combined data suggest the presence of a diverse assemblage of arthropod-associated Bacteroidetes bacteria that are likely to strongly influence their hosts' biology.}, } @article {pmid15181901, year = {2004}, author = {Cerdeño-Tárraga, A and Crossman, L and Parkhill, J}, title = {New environments, versatile genomes.}, journal = {Nature reviews. Microbiology}, volume = {2}, number = {6}, pages = {446-447}, doi = {10.1038/nrmicro910}, pmid = {15181901}, issn = {1740-1526}, mesh = {Environment ; Genomics/*methods ; Phytoplankton/*genetics ; Treponema/*genetics ; Wolbachia/*genetics ; }, } @article {pmid15157232, year = {2004}, author = {Dutton, TJ and Sinkins, SP}, title = {Strain-specific quantification of Wolbachia density in Aedes albopictus and effects of larval rearing conditions.}, journal = {Insect molecular biology}, volume = {13}, number = {3}, pages = {317-322}, doi = {10.1111/j.0962-1075.2004.00490.x}, pmid = {15157232}, issn = {0962-1075}, mesh = {Aedes/*microbiology ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Body Weights and Measures ; DNA Primers ; Electrophoresis, Agar Gel ; Fluorometry ; Larva/microbiology/physiology ; Polymerase Chain Reaction ; Population Density ; Population Dynamics ; Reproduction/genetics ; *Symbiosis ; Wolbachia/*genetics/physiology ; }, abstract = {The density of the endosymbiont Wolbachia can influence the expression of the crossing sterilities known as cytoplasmic incompatibility (CI), and also its rate of maternal transmission. Aedes albopictus mosquitoes contain a superinfection with the Wolbachia strains wAlbA and wAlbB. A strain-specific real-time quantitative PCR assay was developed and used to quantify relative Wolbachia strain densities within individual mosquitoes. The wAlbB strain was consistently found to be at higher density than wAlbA, which can explain a slightly lower rate of maternal transmission reported for wAlbA. The effects of larval crowding and nutritional stress were also examined. Larval crowding always reduced adult size, but reduced the density of Wolbachia strains relative to uncrowded conditions only if crowding was accompanied by restricted nutrient availability. Crowded rearing conditions never resulted in strain segregation or in a reduction in the penetrance of CI, however. The rate of maternal transmission and the penetrance of CI are the two most important variables that determine relative Wolbachia population invasion dynamics, and both are considerably higher here than have been reported in the Drosophila simulans model system.}, } @article {pmid15149411, year = {2004}, author = {Vala, F and Egas, M and Breeuwer, JA and Sabelis, MW}, title = {Wolbachia affects oviposition and mating behaviour of its spider mite host.}, journal = {Journal of evolutionary biology}, volume = {17}, number = {3}, pages = {692-700}, doi = {10.1046/j.1420-9101.2003.00679.x}, pmid = {15149411}, issn = {1010-061X}, mesh = {Animals ; *Biological Evolution ; Cytoplasm/microbiology ; Female ; Male ; Oviposition/*physiology ; Reproduction/physiology ; *Sexual Behavior, Animal ; Tetranychidae/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are transmitted from mother to offspring via the cytoplasm of the egg. When mated to males infected with Wolbachia bacteria, uninfected females produce unviable offspring, a phenomenon called cytoplasmic incompatibility (CI). Current theory predicts that 'sterilization' of uninfected females by infected males confers a fitness advantage to Wolbachia in infected females. When the infection is above a threshold frequency in a panmictic population, CI reduces the fitness of uninfected females below that of infected females and, consequently, the proportion of infected hosts increases. CI is a mechanism that benefits the bacteria but, apparently, not the host. The host could benefit from avoiding incompatible mates. Parasite load and disease resistance are known to be involved in mate choice. Can Wolbachia also be implicated in reproductive behaviour? We used the two-spotted spider mite - Wolbachia symbiosis to address this question. Our results suggest that uninfected females preferably mate to uninfected males while infected females aggregate their offspring, thereby promoting sib mating. Our data agrees with other results that hosts of Wolbachia do not necessarily behave as innocent bystanders - host mechanisms that avoid CI can evolve.}, } @article {pmid15146989, year = {2004}, author = {Hartelt, K and Oehme, R and Frank, H and Brockmann, SO and Hassler, D and Kimmig, P}, title = {Pathogens and symbionts in ticks: prevalence of Anaplasma phagocytophilum (Ehrlichia sp.), Wolbachia sp., Rickettsia sp., and Babesia sp. in Southern Germany.}, journal = {International journal of medical microbiology : IJMM}, volume = {293 Suppl 37}, number = {}, pages = {86-92}, doi = {10.1016/s1433-1128(04)80013-5}, pmid = {15146989}, issn = {1438-4221}, mesh = {Anaplasma phagocytophilum/genetics/*isolation & purification ; Animals ; Citrate (si)-Synthase/chemistry/genetics ; DNA, Bacterial/chemistry/genetics ; Germany/epidemiology ; Humans ; Ixodes/*microbiology ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 16S/chemistry/genetics ; RNA, Ribosomal, 18S/chemistry/genetics ; Rickettsia/genetics/*isolation & purification ; Tick Infestations/epidemiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Tick-transmitted diseases like tick-borne encephalitis and Lyme borreliosis have been well known in Germany for decades. Ongoing research now gives an additional focus to a broad range of other bacteria and parasites in ticks like Anaplasma phagocytophilum, former Ehrlichia sp., Rickettsia sp. and Babesia sp. Knowledge about the prevalence of these infectious agents in ticks is an important prerequisite for risk assessment of human diseases. Therefore nymphs and adult Ixodes ricinus ticks were collected and examined for Anaplasma phagocytophilum (n = 5424 ticks), Rickettsia sp. (n = 1187), and Babesia sp. (n = 3113). For the detection of Anaplasma phagocytophilum, DNA from the 16S rDNA gene was amplified by nested PCR and hybridized with a DIG-labeled oligonucleotide probe. The examination of Rickettsia sp. was performed by single PCR. A partial sequence of the citrate synthase gene was amplified. As a target for the detection of Babesia sp., DNA from the 18S rDNA gene was amplified, also by single PCR. All positive PCR products were sequenced to control specificity. Anaplasma phagocytophilum was detected by PCR in n = 103 (1.9%) out of 5,424 examined ticks from 11 investigation areas. However, not all positive PCR products hybridized using DIG-labeled oligonucleotide probe. Thus, the result of sequencing indicated that only 1.0% (n = 54) belonged to Anaplasma phagocytophilum and nearly half of these PCR products (0.9%) were identified as Wolbachia sp. Rickettsia sp. in Ixodes ricinus ticks from 3 areas were found in n = 105 (8.9%) out of 1,187 ticks examined (range from 13.3% to 5.6%). Sequencing showed Rickettsia helvetica exclusively. In about 2.6% of Rickettsia-positive ticks, double infection with Anaplasma phagocytophilum was found. Babesia sp. was detected in n= 31 (1.0%) out of 3,113 ticks examined, which originated from 4 different areas. By sequencing, n = 28 (90.0%) were identified as Babesia divergens. Three of all Babesia-positive ticks were identified as harboring Babesia microti. The detection of Anaplasma phagocytophilum, Rickettsia sp. and Babesia sp. demonstrates their possible role as a source of human infection in Germany.}, } @article {pmid15140268, year = {2004}, author = {Perotti, MA and Catalá, SS and Ormeño, Adel V and Zelazowska, M and Biliński, SM and Braig, HR}, title = {The sex ratio distortion in the human head louse is conserved over time.}, journal = {BMC genetics}, volume = {5}, number = {}, pages = {10}, pmid = {15140268}, issn = {1471-2156}, mesh = {Africa/epidemiology ; Animals ; Anoplura/microbiology ; Argentina/epidemiology ; Asia/epidemiology ; Australia/epidemiology ; Child ; Female ; Humans ; Israel/epidemiology ; Lice Infestations/*epidemiology/parasitology/veterinary ; Male ; Molecular Sequence Data ; North America/epidemiology ; Pediculus/microbiology/*physiology ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rural Population ; Schools ; *Sex Ratio ; Species Specificity ; Swine Diseases/parasitology ; Time Factors ; United Kingdom/epidemiology ; Urban Population ; Wolbachia/genetics/isolation & purification ; }, abstract = {BACKGROUND: At the turn of the 19th century the first observations of a female-biased sex ratio in broods and populations of the head louse, Pediculus humanus capitis, had been reported. A study by Buxton in 1940 on the sex ratio of lice on prisoners in Ceylon is still today the subject of reanalyses. This sex ratio distortion had been detected in ten different countries. In the last sixty years no new data have been collected, especially on scalp infestations under economically and socially more developed conditions.

RESULTS: Here we report a female bias of head lice in a survey of 480 school children in Argentina. This bias is independent of the intensity of the pediculosis, which makes local mate competition highly unlikely as the source of the aberrant sex ratio; however, other possible adaptive mechanisms cannot be discounted. These lice as well as lice from pupils in Britain were carrying several strains of the endosymbiotic bacterium Wolbachia pipientis, one of the most wide spread intracellular sex ratio distorters. Similar Wolbachia strains are also present in the pig louse, Haematopinus suis, suggesting that this endosymbiont might have a marked influence on the biology of the whole order. The presence of a related obligate nutritional bacterium in lice prevents the investigation of a causal link between sex ratio and endosymbionts.

CONCLUSIONS: Regardless of its origin, this sex ratio distortion in head lice that has been reported world wide, is stable over time and is a remarkable deviation from the stability of frequency-dependent selection of Fisher's sex ratio. A female bias first reported in 1898 is still present over a hundred years and a thousand generations later.}, } @article {pmid15138452, year = {2004}, author = {Cordaux, R and Michel-Salzat, A and Frelon-Raimond, M and Rigaud, T and Bouchon, D}, title = {Evidence for a new feminizing Wolbachia strain in the isopod Armadillidium vulgare: evolutionary implications.}, journal = {Heredity}, volume = {93}, number = {1}, pages = {78-84}, doi = {10.1038/sj.hdy.6800482}, pmid = {15138452}, issn = {0018-067X}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; *Biological Evolution ; DNA, Mitochondrial/genetics ; Genetic Markers ; Genetics, Population ; Isopoda/*genetics ; Phenotype ; Phylogeny ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia are intracellular maternally inherited alpha-Proteobacteria infecting a wide range of arthropods. In the common pill bug Armadillidium vulgare, the known Wolbachia strain is responsible for feminization of genetic males. We have investigated Wolbachia diversity in 20 populations of A. vulgare from west and east Europe, north Africa and north America. A new Wolbachia strain (wVulM) was identified through the variability of the wsp gene, distantly related to that previously known (wVulC) in this host species. No individual with multiple infections was detected. Inoculation experiments indicated that the new wVulM bacterial strain also induces feminization in A. vulgare. However, the wVulC strain showed a higher transmission rate than the wVulM strain and was the most geographically widespread Wolbachia in A. vulgare populations. Mitochondrial 16SrDNA gene sequencing was conducted in Wolbachia-infected individuals, revealing the occurrence of four host lineages. The comparison of bacterial strains and their respective host mitochondrial phylogenies failed to show concordance, indicating horizontal transmission of the Wolbachia strains within populations of A. vulgare.}, } @article {pmid15129961, year = {2004}, author = {Huigens, ME and de Almeida, RP and Boons, PA and Luck, RF and Stouthamer, R}, title = {Natural interspecific and intraspecific horizontal transfer of parthenogenesis-inducing Wolbachia in Trichogramma wasps.}, journal = {Proceedings. Biological sciences}, volume = {271}, number = {1538}, pages = {509-515}, pmid = {15129961}, issn = {0962-8452}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; DNA Primers ; *Disease Transmission, Infectious ; Female ; Larva/microbiology ; Parthenogenesis/*physiology ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA ; Species Specificity ; Wasps/*microbiology/physiology ; Wolbachia/*genetics ; }, abstract = {The intracellular bacterium Wolbachia is one of the most common symbionts in arthropods and, because of its manipulative effects on host reproduction, is assumed to be an important factor in several evolutionary processes. These bacteria are mainly vertically transmitted from mother to daughter through the egg cytoplasm, and horizontal transmission is generally assumed to be rare. Here, we show natural inter- and intraspecific horizontal transfer of parthenogenesis-inducing Wolbachia between parasitoid wasps of the genus Trichogramma. Horizontal transfer was observed when infected and uninfected larvae shared the same host egg. This is the first report, to our knowledge, on interspecific horizontal transfer of Wolbachia between closely related sympatric species. Some originally uninfected immature wasps acquired Wolbachia while inside the host egg, but not all of these newly infected females exhibited the parthenogenesis phenotype. In general, intraspecific horizontal transfer was more successful than interspecific transfer. Wolbachia underwent vertical transmission in the new species but the infection tended to be lost within several generations. Our results have important implications for understanding the evolution of Wolbachia-host associations.}, } @article {pmid15127087, year = {2004}, author = {Dobson, SL and Rattanadechakul, W and Marsland, EJ}, title = {Fitness advantage and cytoplasmic incompatibility in Wolbachia single- and superinfected Aedes albopictus.}, journal = {Heredity}, volume = {93}, number = {2}, pages = {135-142}, doi = {10.1038/sj.hdy.6800458}, pmid = {15127087}, issn = {0018-067X}, support = {AI051533/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*physiology ; Analysis of Variance ; Animals ; *Biological Evolution ; Crosses, Genetic ; Cytoplasm/microbiology ; Female ; Fertility/physiology ; *Genetics, Population ; Genotype ; Longevity ; Models, Biological ; Polymerase Chain Reaction ; Reproduction/physiology ; *Selection, Genetic ; *Symbiosis ; Time Factors ; *Wolbachia ; }, abstract = {Wolbachia are obligate, maternally inherited, intracellular bacteria that infect numerous insects and other invertebrates. Wolbachia infections have evolved multiple mechanisms to manipulate host reproduction and facilitate invasion of naive host populations. One such mechanism is cytoplasmic incompatibility (CI) that occurs in many insect species, including Aedes albopictus (Asian tiger mosquito). The multiple Wolbachia infections that occur naturally in A. albopictus make this mosquito a useful system in which to study CI. Here, experiments employ mosquito strains that have been introgressed to provide genetically similar strains that harbor differing Wolbachia infection types. Cytoplasmic incompatibility levels, host longevity, egg hatch rates, and fecundity are examined. Crossing results demonstrate a pattern of additive unidirectional cytoplasmic incompatibility. Furthermore, relative to uninfected females, infected females are at a reproductive advantage due to both cytoplasmic incompatibility and a fitness increase associated with Wolbachia infection. In contrast, no fitness difference was observed in comparisons of single- and superinfected females. We discuss the observed results in regard to the evolution of the Wolbachia/A. albopictus symbiosis and the observed pattern of Wolbachia infection in natural populations.}, } @article {pmid15111096, year = {2004}, author = {Rodgers-Gray, TP and Smith, JE and Ashcroft, AE and Isaac, RE and Dunn, AM}, title = {Mechanisms of parasite-induced sex reversal in Gammarus duebeni.}, journal = {International journal for parasitology}, volume = {34}, number = {6}, pages = {747-753}, doi = {10.1016/j.ijpara.2004.01.005}, pmid = {15111096}, issn = {0020-7519}, mesh = {Amphipoda/anatomy & histology/*parasitology/physiology ; Androgens/physiology ; Animals ; Female ; Gametogenesis/physiology ; Gonadal Hormones/physiology ; *Hermaphroditic Organisms ; Host-Parasite Interactions/physiology ; Ions/metabolism ; Male ; Nosema/*physiology ; Ovary/anatomy & histology/physiology ; *Sex Determination Processes ; Sex Differentiation/physiology ; Testis/anatomy & histology/physiology ; Vas Deferens/anatomy & histology/physiology ; }, abstract = {The amphipod Gammarus duebeni is host to the feminising microsporidian parasite Nosema granulosis that converts males into functional females. To test the hypothesis that the parasite acts through endocrine disruption we compared the morphology of the gonad and activity of the androgenic gland, which coordinates male sexual differentiation, in infected and uninfected animals. Male gonad consisted of testis, seminal vesicle and vas deferens that was anchored to the genital papilla on segment 7. The androgenic gland was associated with the distal end of the vas deferens. In female and intersex animals the bi-lobed ovary opened into the oviduct at segment 5, vestigial vas deferens and vestigial androgenic gland were retained. The majority of parasitised individuals (38/39) were either phenotypic females or intersexes with fully developed ovaries and an undifferentiated androgenic gland. Our data suggest that the parasite prevents differentiation of the androgenic gland. In further support of this hypothesis, mass spectrometry of a single androgenic gland from males revealed a dominant molecular ion with a mass/charge ratio of 4818.4+H, corresponding to a peptide of androgenic gland hormone from Armadillidium vulgare. In contrast the vestigial androgenic gland from parasitised and unparasitised females showed only low intensity peaks. Our observations demonstrate that the parasite manipulates host sex by preventing androgenic gland differentiation, androgenic gland hormone production and consequently male differentiation. This is in agreement with observations of A. vulgare with inherited Wolbachia infection, suggesting that phylogenetically distant feminisers manipulate hosts through a common mechanism. The high frequency of infection in intersexes (89.3%) suggests that this phenotype results from incomplete feminisation by the parasite.}, } @article {pmid15111095, year = {2004}, author = {Foster, JM and Kumar, S and Ganatra, MB and Kamal, IH and Ware, J and Ingram, J and Pope-Chappell, J and Guiliano, D and Whitton, C and Daub, J and Blaxter, ML and Slatko, BE}, title = {Construction of bacterial artificial chromosome libraries from the parasitic nematode Brugia malayi and physical mapping of the genome of its Wolbachia endosymbiont.}, journal = {International journal for parasitology}, volume = {34}, number = {6}, pages = {733-746}, doi = {10.1016/j.ijpara.2004.02.001}, pmid = {15111095}, issn = {0020-7519}, mesh = {Animals ; Base Sequence ; Brugia malayi/*genetics ; Chromosome Mapping/*methods ; Chromosomes, Artificial, Bacterial/*genetics ; Contig Mapping/methods ; DNA, Bacterial/genetics ; DNA, Protozoan/genetics ; Genome, Bacterial ; Genome, Protozoan ; Genomic Library ; Molecular Weight ; Plasmids ; RNA, Bacterial/genetics ; RNA, Ribosomal/genetics ; Restriction Mapping/methods ; Sequence Analysis, DNA/methods ; Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {The parasitic nematode, Brugia malayi, causes lymphatic filariasis in humans, which in severe cases leads to the condition known as elephantiasis. The parasite contains an endosymbiotic alpha-proteobacterium of the genus Wolbachia that is required for normal worm development and fecundity and is also implicated in the pathology associated with infections by these filarial nematodes. Bacterial artificial chromosome libraries were constructed from B. malayi DNA and provide over 11-fold coverage of the nematode genome. Wolbachia genomic fragments were simultaneously cloned into the libraries giving over 5-fold coverage of the 1.1 Mb bacterial genome. A physical framework for the Wolbachia genome was developed by construction of a plasmid library enriched for Wolbachia DNA as a source of sequences to hybridise to high-density bacterial artificial chromosome colony filters. Bacterial artificial chromosome end sequencing provided additional Wolbachia probe sequences to facilitate assembly of a contig that spanned the entire genome. The Wolbachia sequences provided a marker approximately every 10 kb. Four rare-cutting restriction endonucleases were used to restriction map the genome to a resolution of approximately 60 kb and demonstrate concordance between the bacterial artificial chromosome clones and native Wolbachia genomic DNA. Comparison of Wolbachia sequences to public databases using BLAST algorithms under stringent conditions allowed confident prediction of 69 Wolbachia peptide functions and two rRNA genes. Comparison to closely related complete genomes revealed that while most sequences had orthologs in the genome of the Wolbachia endosymbiont from Drosophila melanogaster, there was no evidence for long-range synteny. Rather, there were a few cases of short-range conservation of gene order extending over regions of less than 10 kb. The molecular scaffold produced for the genome of the Wolbachia from B. malayi forms the basis of a genomic sequencing effort for this bacterium, circumventing the difficult challenge of purifying sufficient endosymbiont DNA from a tropical parasite for a whole genome shotgun sequencing strategy.}, } @article {pmid15094394, year = {2004}, author = {Fujii, Y and Kubo, T and Ishikawa, H and Sasaki, T}, title = {Isolation and characterization of the bacteriophage WO from Wolbachia, an arthropod endosymbiont.}, journal = {Biochemical and biophysical research communications}, volume = {317}, number = {4}, pages = {1183-1188}, doi = {10.1016/j.bbrc.2004.03.164}, pmid = {15094394}, issn = {0006-291X}, mesh = {Animals ; Bacteriophages/genetics/*isolation & purification/ultrastructure ; DNA/isolation & purification ; Electrophoresis, Agar Gel ; Genome, Bacterial ; Lysogeny ; Microscopy, Electron ; Moths/*microbiology ; Open Reading Frames ; Symbiosis ; Viral Proteins/genetics ; Wolbachia/genetics/physiology/*virology ; }, abstract = {Wolbachia is a group of obligate symbiotic bacteria found in many insects and other arthropods. The presence of Wolbachia alters reproduction in the host, but the mechanisms are unknown. Molecular biological studies of Wolbachia have delayed significantly, and one of the reasons is the lack of transformation techniques of this bacterium. In the present study, bacteriophage particles were isolated from Wolbachia for the first time. The purified phage had an isometric head that was approximately 40 nm in diameter and contained linear double-stranded DNA of approximately 20 kbp. Partial sequence information (total of 20,484 bp) revealed that there were 24 open reading frames including a structural gene module, and genes for replication and lysogenic conversion. This bacteriophage is the only known mobile genetic element potentially used for transformation of Wolbachia.}, } @article {pmid15088646, year = {2004}, author = {Merçot, H and Charlat, S}, title = {Wolbachia infections in Drosophila melanogaster and D. simulans: polymorphism and levels of cytoplasmic incompatibility.}, journal = {Genetica}, volume = {120}, number = {1-3}, pages = {51-59}, doi = {10.1023/b:gene.0000017629.31383.8f}, pmid = {15088646}, issn = {0016-6707}, mesh = {Animals ; Cytoplasm/*metabolism ; Drosophila/*microbiology ; Drosophila melanogaster/*microbiology ; Female ; Male ; Polymorphism, Genetic ; Rickettsiaceae Infections/*pathology ; Sex Factors ; Species Specificity ; Wolbachia/*metabolism ; }, abstract = {Wolbachia are endosymbiotic bacteria, widespread in terrestrial Arthropods. They are mainly transmitted vertically, from mothers to offspring and induce various alterations of their hosts' sexuality and reproduction, the most commonly reported phenomenon being Cytoplasmic Incompatibility (CI), observed in Drosophila melanogaster and D. simulans. Basically, CI results in a more or less intense embryonic mortality, occurring in crosses between males infected by Wolbachia and uninfected females. In D. simulans, Wolbachia and CI were observed in 1986. Since then, this host species has become a model system for investigating the polymorphism of Wolbachia infections and CI. In this review we describe the different Wolbachia infections currently known to occur in D. melanogaster and D. simulans. The two species are highly contrasting with regard to symbiotic diversity: while five Wolbachia variants have been described in D. simulans natural populations, D. melanogaster seems to harbor one Wolbachia variant only. Another marked difference between these two Drosophila species is their permissiveness with regard to CI, which seems to be fully expressed in D. simulans but partially or totally repressed in D. melanogaster, demonstrating the involvement of host factors in the control of CI levels. The potential of the two host species regarding the understanding of CI and its evolution is also discussed.}, } @article {pmid15088645, year = {2004}, author = {Solignac, M}, title = {Mitochondrial DNA in the Drosophila melanogaster complex.}, journal = {Genetica}, volume = {120}, number = {1-3}, pages = {41-50}, doi = {10.1023/b:gene.0000017628.52056.97}, pmid = {15088645}, issn = {0016-6707}, mesh = {Animals ; Cell Nucleus/metabolism ; Cytoplasm/metabolism ; DNA, Mitochondrial/*genetics/metabolism ; Drosophila ; Drosophila melanogaster/*genetics ; Genome ; Haplotypes ; Phylogeny ; Polymorphism, Genetic ; Species Specificity ; Wolbachia ; }, abstract = {Mitochondrial DNA in the complex Drosophila melanogaster was among the first studied in metazoans. The variability of the molecule was extensively studied using restriction enzymes, gene sequence and recently sequence of the whole coding region. Within the complex, seven major haplotypes have been described, one (me) in D. melanogaster, three in D. simulans (siI, siII, siIII), two in D. mauritiana (maI, maII), and one in D. sechellia (se). The molecular distance between the haplotypes is comprised between 1 and 5%, except for siII and maI, which are virtually identical. The nucleotide diversity within each of these haplotypes is very low, varying from 0 to 0.0005. Most of the cytoplasms are infected by the bacterium Wolbachia and different bacterial strains infect cytoplasms harboring different mtDNA types. mtDNA polymorphism is discussed in relation with Wolbachia, nuclear polymorphism and speciation events.}, } @article {pmid15088643, year = {2004}, author = {Capy, P and Gibert, P}, title = {Drosophila melanogaster, Drosophila simulans: so similar yet so different.}, journal = {Genetica}, volume = {120}, number = {1-3}, pages = {5-16}, doi = {10.1023/b:gene.0000017626.41548.97}, pmid = {15088643}, issn = {0016-6707}, mesh = {Animals ; Cell Movement ; Drosophila/*genetics/physiology ; Drosophila melanogaster/*genetics/physiology ; Genetic Variation ; Phenotype ; Polymorphism, Genetic ; Species Specificity ; }, abstract = {During the last two decades, the two cosmopolitan species Drosophila melanogaster and Drosophila simulans have been compared with regard to numerous characteristics, ranging from their geographic distribution and ecology to their DNA polymorphism. Various traits have been compared, including morphology, physiology, sexual behavior, allozymes and other proteins, chromosomal inversions, mitochondrial and nuclear DNA, transposable elements, wolbachia etc. Such comparisons reveal similarities and differences between the two species, depending on the trait considered. In most cases, the between-population variability of D. simulans is lower than that of D. melanogaster, but the two species exhibit similar levels of within-population variability. One of the main exceptions is the nucleotide polymorphism of several nuclear regions. Although several hypotheses have been proposed to explain these observations, the evolutionary dynamics of these two species are far from being understood. How have two species sharing a common ancestor in the recent past accumulated so many differences? A brief history of comparisons of the two species, from the first in 1919 by A.H. Sturtevant, and a summary of the hypotheses proposed to explain the similarities and the differences between these species are presented and discussed.}, } @article {pmid15087494, year = {2004}, author = {Dyson, EA and Hurst, GD}, title = {Persistence of an extreme sex-ratio bias in a natural population.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {101}, number = {17}, pages = {6520-6523}, pmid = {15087494}, issn = {0027-8424}, mesh = {Animals ; Butterflies ; Female ; Male ; Molecular Sequence Data ; Population Dynamics ; Samoa ; *Sex Ratio ; }, abstract = {The sex ratio is a key parameter in the evolution and ecology of a species. Selfish genetic elements that bias the sex ratio of affected individuals are well known and characterized, but their effect on populations has been considered limited, because either the element does not achieve high prevalence or the host rapidly evolves resistance to the distorting element, reducing its prevalence. We tested whether the host necessarily prevails by using a butterfly system where records from the early part of the 20th century reported extreme sex-ratio bias in nature. We reexamined this population and found the bias was present today, 400 generations after the original record, with a population sex ratio of 100 females per male. The sex-ratio bias was associated with the presence of a heritable male-killing Wolbachia infection in 99% of adult females, against which the host butterfly has failed to evolve resistance. The resultant dearth of males causes an average 57% reduction in the reproductive output of adult females. Persistence of the population despite the very high frequency of the sex-ratio distorter appears to be associated with the ability of males to mate >50 times in their life combined with a high intrinsic rate of increase of the species.}, } @article {pmid15078450, year = {2004}, author = {Pannebakker, BA and Zwaan, BJ and Beukeboom, LW and Van Alphen, JJ}, title = {Genetic diversity and Wolbachia infection of the Drosophila parasitoid Leptopilina clavipes in western Europe.}, journal = {Molecular ecology}, volume = {13}, number = {5}, pages = {1119-1128}, doi = {10.1111/j.1365-294X.2004.02147.x}, pmid = {15078450}, issn = {0962-1083}, mesh = {Animals ; Cluster Analysis ; Drosophila/*parasitology ; Europe ; *Genetic Variation ; Genetics, Population ; Genotype ; Parthenogenesis/*physiology ; Polymorphism, Restriction Fragment Length ; Wasps/*genetics/*microbiology/physiology ; *Wolbachia ; }, abstract = {Wolbachia are maternally transmitted bacteria that alter their arthropod hosts' reproduction in various ways, including parthenogenesis induction (PI). Wolbachia-induced parthenogenesis can have drastic effects on the genetic structure of its host because it potentially reduces populations to clones without genetic exchange. However, Wolbachia-induced parthenogenesis does not inevitably result in a reduction of genetic variation of infected populations vs. uninfected populations, because the parthenogenetic populations are initially derived from uninfected populations and can thus show similar genetic variation. Here we investigate these issues in infected and uninfected populations of the Drosophila parasitoid Leptopilina clavipes in western Europe. Wasps from 19 sites in the Netherlands, France and northern Spain were screened for Wolbachia and analysed using amplified fragment length polymorphism (AFLP) markers. All the populations from the Netherlands and mid-France were infected with the same two strains of Wolbachia, whereas populations from the Pyrenees were not infected. The infected and uninfected populations show identical levels of genetic variation, but have clearly diverged genetically, indicating the presence of a barrier that prevents gene flow. Within the infected wasps two distinct genotypes were found at multiple localities, indicating the coexistence of multiple clones. The conditions promoting clonal coexistence in L. clavipes are discussed.}, } @article {pmid15061286, year = {2004}, author = {Rasgon, JL and Scott, TW}, title = {An initial survey for Wolbachia (Rickettsiales: Rickettsiaceae) infections in selected California mosquitoes (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {41}, number = {2}, pages = {255-257}, doi = {10.1603/0022-2585-41.2.255}, pmid = {15061286}, issn = {0022-2585}, mesh = {Animals ; California ; Culicidae/*microbiology ; Wolbachia/classification/*isolation & purification ; }, abstract = {Knowledge of biogeographic variation in Wolbachia infection rates and inferred susceptibility to infection among different mosquito taxa has fundamental implications for the design and successful application of Wolbachia-based vector-borne disease control strategies. Using a Wolbachia-specific polymerase chain reaction assay, we tested 14 North American mosquito species in five genera (Aedes, Anopheles, Culiseta, Culex, and Ochlerotatus) for Wolbachia infection. Wolbachia infections were only detected in members of the Culex pipiens (L.) species complex.}, } @article {pmid15060582, year = {2004}, author = {Pannebakker, BA and Pijnacker, LP and Zwaan, BJ and Beukeboom, LW}, title = {Cytology of Wolbachia-induced parthenogenesis in Leptopilina clavipes (Hymenoptera: Figitidae).}, journal = {Genome}, volume = {47}, number = {2}, pages = {299-303}, doi = {10.1139/g03-137}, pmid = {15060582}, issn = {0831-2796}, mesh = {Animals ; Diploidy ; Meiosis ; Mitosis ; Parthenogenesis/*genetics/physiology ; Wasps/*cytology/genetics/*microbiology ; Wolbachia/*physiology ; }, abstract = {Parthenogenesis induced by cytoplasmatically inherited Wolbachia bacteria has been found in a number of arthropod species, mainly Hymenoptera. Previously, two different forms of diploidy restoration have been reported to underlie parthenogenesis induction in Hymenoptera by Wolbachia. Both are a form of gamete duplication, but each differs in their timing. We investigated the cytology of the early embryonic development of a Wolbachia-infected strain of the parasitoid wasp Leptopilina clavipes and compared it with that of an uninfected sexual strain. Both strains have a similar meiosis. In the infected parthenogenetic strain, diploidy is restored by anaphase restitution during the first somatic mitosis, similar to Trichogramma, but not to Muscidifurax. Our results confirm the occurrence of different cytological mechanisms of diploidy restoration associated with parthenogenesis-inducing Wolbachia in the order Hymenoptera.}, } @article {pmid15058435, year = {2004}, author = {Kageyama, D and Traut, W}, title = {Opposite sex-specific effects of Wolbachia and interference with the sex determination of its host Ostrinia scapulalis.}, journal = {Proceedings. Biological sciences}, volume = {271}, number = {1536}, pages = {251-258}, pmid = {15058435}, issn = {0962-8452}, mesh = {Animals ; DNA Primers ; Fluorescence ; Japan ; Karyotyping ; Moths/*genetics/*microbiology ; Polymerase Chain Reaction ; Sex Chromosomes/*genetics ; *Sex Determination Processes ; Sex Factors ; *Sex Ratio ; Tetracycline ; Transfection ; Wolbachia/*physiology ; }, abstract = {In the adzuki bean borer, Ostrinia scapulalis, the sex ratio in most progenies is 1 : 1. Females from Wolbachia-infected matrilines, however, give rise to all-female broods when infected and to all-male broods when cured of the infection. These observations had been interpreted as Wolbachia-induced feminization of genetic males into functional females. Here, we show that the interpretation is incorrect. Females from both lines have a female karyotype with a WZ sex-chromosome constitution while males are ZZ. At the time of hatching from eggs, WZ and ZZ individuals are present at a 1 : 1 ratio in broods from uninfected, infected and cured females. In broods from Wolbachia-infected females, ZZ individuals die during larval development, whereas in those from cured females, WZ individuals die. Hence, development of ZZ individuals is impaired by Wolbachia but development of WZ females may require the presence of Wolbachia in infected matrilines. Sexual mosaics generated (i) by transfection of uninfected eggs and (ii) by tetracycline treatment of Wolbachia-infected mothers prior to oviposition were ZZ in all tissues, including typically female organs. We conclude that: (i) Wolbachia acts by manipulating the sex determination of its host; and (ii) although sexual mosaics can survive, development of a normal female is incompatible with a ZZ genotype.}, } @article {pmid15056362, year = {2004}, author = {Gavotte, L and Vavre, F and Henri, H and Ravallec, M and Stouthamer, R and Boulétreau, M}, title = {Diversity, distribution and specificity of WO phage infection in Wolbachia of four insect species.}, journal = {Insect molecular biology}, volume = {13}, number = {2}, pages = {147-153}, doi = {10.1111/j.0962-1075.2004.00471.x}, pmid = {15056362}, issn = {0962-1075}, mesh = {Animals ; Bacteriophages/*genetics/ultrastructure ; Base Sequence ; Capsid Proteins/genetics ; DNA Primers ; Drosophila/microbiology/*parasitology ; Geography ; Microscopy, Electron ; Molecular Sequence Data ; *Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; Wasps/*microbiology ; Wolbachia/genetics/*virology ; }, abstract = {The bacteriophage WO was recently characterized in Wolbachia, a strictly intracellular bacterium that causes several reproductive alterations in its arthropod hosts. To gain insights into the phage-Wolbachia relationships, we studied the phage presence among Wolbachia infecting four insect species sharing several Wolbachia strains, two Drosophila and two of their parasitoid wasps. Based on the phage sequence of ORF7, we identified five different phages in six Wolbachia strains. Among these five bacteriophages, some are specific for a given bacterial strain whereas others are not, but globally phage infection appears stable on a large geographical scale and across insect generations. Their specificity contrasts with the absence of congruence between Wolbachia and phage phylogenies, suggesting phage exchanges between different Wolbachia lineages.}, } @article {pmid15049082, year = {2003}, author = {Hoti, SL and Sridhar, A and Das, PK}, title = {Presence of Wolbachia endosymbionts in microfilariae of Wuchereria bancrofti (Spirurida: Onchocercidae) from different geographical regions in India.}, journal = {Memorias do Instituto Oswaldo Cruz}, volume = {98}, number = {8}, pages = {1017-1019}, doi = {10.1590/s0074-02762003000800006}, pmid = {15049082}, issn = {0074-0276}, mesh = {Animals ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; India ; Microfilariae/*microbiology ; Polymerase Chain Reaction ; Wolbachia/*isolation & purification ; Wuchereria bancrofti/*microbiology ; }, abstract = {In view of the recent discovery of rickettsial endosymbionts, Wolbachia in lymphatic filarial parasites, Wuchereria bancrofti and Brugia malayi and subsequently of their vital role in the survival and development of the latter, antibiotics such as tetracycline are being suggested for the treatment of lymphatic filariasis, by way of eliminating the endosymbiont. But, it is essential to assess their presence in parasites from areas endemic for lymphatic filariasis before such a new control tool is employed. In the present communication, we report the detection of Wolbachia endosymbionts in microfilariae of W. bancrofti parasites collected from geographically distant locations of India, such as Pondicherry (Union Territory), Calicut (Kerala), Jagadalpur (Madhya Pradesh), Thirukoilur (TamilNadu), Chinnanergunam (TamilNadu), Rajahmundry (Andhra Pradesh), and Varanasi (Uttar Pradesh), using Wolbachia specific 16S rDNA polymerase chain reaction.}, } @article {pmid15037105, year = {2004}, author = {Casiraghi, M and Bain, O and Guerrero, R and Martin, C and Pocacqua, V and Gardner, SL and Franceschi, A and Bandi, C}, title = {Mapping the presence of Wolbachia pipientis on the phylogeny of filarial nematodes: evidence for symbiont loss during evolution.}, journal = {International journal for parasitology}, volume = {34}, number = {2}, pages = {191-203}, doi = {10.1016/j.ijpara.2003.10.004}, pmid = {15037105}, issn = {0020-7519}, mesh = {Animals ; Base Sequence ; DNA, Mitochondrial/analysis ; Filarioidea/genetics/*parasitology ; Genes, Bacterial ; Genes, Helminth ; Host-Parasite Interactions ; Molecular Sequence Data ; *Phylogeny ; Polymerase Chain Reaction/methods ; Symbiosis ; Wolbachia/*classification/genetics ; }, abstract = {Wolbachia pipientis is a bacterial endosymbiont associated with arthropods and filarial nematodes. In filarial nematodes, W. pipientis has been shown to play an important role in the biology of the host and in the immuno-pathology of filariasis. Several species of filariae, including the most important parasites of humans and animals (e.g. Onchocerca volvulus, Wuchereria bancrofti and Dirofilaria immitis) have been shown to harbour these bacteria. Other filarial species, including an important rodent species (Acanthocheilonema viteae), which has been used as a model for the study of filariasis, do not appear to harbour these symbionts. There are still several open questions about the distribution of W. pipientis in filarial nematodes. Firstly the number of species examined is still limited. Secondly, it is not clear whether the absence of W. pipientis in negative species could represent an ancestral characteristic or the result of a secondary loss. Thirdly, several aspects of the phylogeny of filarial nematodes are still unclear and it is thus difficult to overlay the presence/absence of W. pipientis on a tree representing filarial evolution. Here we present the results of a PCR screening for W. pipientis in 16 species of filariae and related nematodes, representing different families/subfamilies. Evidence for the presence of W. pipientis is reported for five species examined for the first time (representing the genera Litomosoides, Litomosa and Dipetalonema); original results on the absence of this bacterium are reported for nine species; for the remaining two species, we have confirmed the absence of W. pipientis recently reported by other authors. In the positive species, the infecting W. pipientis bacteria have been identified through 16S rDNA gene sequence analysis. In addition to the screening for W. pipientis in 16 species, we have generated phylogenetic reconstructions based on mitochondrial gene sequences (12S rDNA; COI), including a total of 28 filarial species and related spirurid nematodes. The mapping of the presence/absence of W. pipientis on the trees generated indicates that these bacteria have possibly been lost during evolution along some lineages of filarial nematodes.}, } @article {pmid15036143, year = {2004}, author = {McGraw, EA and O'Neill, SL}, title = {Wolbachia pipientis: intracellular infection and pathogenesis in Drosophila.}, journal = {Current opinion in microbiology}, volume = {7}, number = {1}, pages = {67-70}, doi = {10.1016/j.mib.2003.12.003}, pmid = {15036143}, issn = {1369-5274}, mesh = {Animals ; Drosophila/*immunology/*microbiology ; Host-Parasite Interactions/immunology ; Virulence ; Wolbachia/*immunology/*pathogenicity ; }, abstract = {Wolbachia pipientis is a vertically transmitted, obligate intracellular symbiont of arthropods. The bacterium is best known for its ability to manipulate host reproductive biology where it can induce cytoplasmic incompatibility, parthenogenesis, feminization and male-killing. In addition to the various reproductive phenotypes it generates through interaction with host reproductive tissue it is also known to infect somatic tissues. However, relatively little is known about the consequences of infection of these tissues with the exception that in some hosts Wolbachia acts as a classical mutualist and in others a pathogen, dramatically shortening adult insect lifespan. Manipulation experiments have demonstrated that the severity of Wolbachia-induced effects on the host is determined by a combination of host genotype, Wolbachia strain, host tissue localization, and interaction with the environment. The recent completion of the whole genome sequence of Wolbachia pipientis wMel strain indicates that it is likely to use a type IV secretion system to establish and maintain infection in its host. Moreover, an unusual abundance of genes encoding proteins with eukaryotic-like ankyrin repeat domains suggest a function in the various described phenotypic effects in hosts.}, } @article {pmid15024419, year = {2004}, author = {Wu, M and Sun, LV and Vamathevan, J and Riegler, M and Deboy, R and Brownlie, JC and McGraw, EA and Martin, W and Esser, C and Ahmadinejad, N and Wiegand, C and Madupu, R and Beanan, MJ and Brinkac, LM and Daugherty, SC and Durkin, AS and Kolonay, JF and Nelson, WC and Mohamoud, Y and Lee, P and Berry, K and Young, MB and Utterback, T and Weidman, J and Nierman, WC and Paulsen, IT and Nelson, KE and Tettelin, H and O'Neill, SL and Eisen, JA}, title = {Phylogenomics of the reproductive parasite Wolbachia pipientis wMel: a streamlined genome overrun by mobile genetic elements.}, journal = {PLoS biology}, volume = {2}, number = {3}, pages = {E69}, pmid = {15024419}, issn = {1545-7885}, support = {UO1-AI47409-01/AI/NIAID NIH HHS/United States ; }, mesh = {Adenosine Triphosphate/chemistry ; Animals ; Cell Lineage ; DNA/chemistry/genetics ; DNA Primers/chemistry ; Drosophila melanogaster/microbiology ; Evolution, Molecular ; Gene Deletion ; Gene Duplication ; Gene Library ; Genes, Bacterial ; Genome ; Genome, Bacterial ; Genomics/*methods ; Glycolysis ; Interspersed Repetitive Sequences ; Models, Genetic ; Molecular Sequence Data ; Open Reading Frames ; Parasites ; Phylogeny ; Polymerase Chain Reaction ; Protein Structure, Tertiary ; Purines/chemistry ; Wolbachia/*genetics ; }, abstract = {The complete sequence of the 1,267,782 bp genome of Wolbachia pipientis wMel, an obligate intracellular bacteria of Drosophila melanogaster, has been determined. Wolbachia, which are found in a variety of invertebrate species, are of great interest due to their diverse interactions with different hosts, which range from many forms of reproductive parasitism to mutualistic symbioses. Analysis of the wMel genome, in particular phylogenomic comparisons with other intracellular bacteria, has revealed many insights into the biology and evolution of wMel and Wolbachia in general. For example, the wMel genome is unique among sequenced obligate intracellular species in both being highly streamlined and containing very high levels of repetitive DNA and mobile DNA elements. This observation, coupled with multiple evolutionary reconstructions, suggests that natural selection is somewhat inefficient in wMel, most likely owing to the occurrence of repeated population bottlenecks. Genome analysis predicts many metabolic differences with the closely related Rickettsia species, including the presence of intact glycolysis and purine synthesis, which may compensate for an inability to obtain ATP directly from its host, as Rickettsia can. Other discoveries include the apparent inability of wMel to synthesize lipopolysaccharide and the presence of the most genes encoding proteins with ankyrin repeat domains of any prokaryotic genome yet sequenced. Despite the ability of wMel to infect the germline of its host, we find no evidence for either recent lateral gene transfer between wMel and D. melanogaster or older transfers between Wolbachia and any host. Evolutionary analysis further supports the hypothesis that mitochondria share a common ancestor with the alpha-Proteobacteria, but shows little support for the grouping of mitochondria with species in the order Rickettsiales. With the availability of the complete genomes of both species and excellent genetic tools for the host, the wMel-D. melanogaster symbiosis is now an ideal system for studying the biology and evolution of Wolbachia infections.}, } @article {pmid15024418, year = {2004}, author = {Wernegreen, JJ}, title = {Endosymbiosis: lessons in conflict resolution.}, journal = {PLoS biology}, volume = {2}, number = {3}, pages = {E68}, pmid = {15024418}, issn = {1545-7885}, support = {R01 GM062626/GM/NIGMS NIH HHS/United States ; R01 GM62626-01/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Evolution, Molecular ; Genome ; Insecta/microbiology ; Parasites ; *Symbiosis ; Wolbachia ; }, abstract = {Endosymbiotic bacteria live within a host species. There are many and diverse examples of such relationships, the study of which provides important lessons for ecology and evolution}, } @article {pmid15009266, year = {2004}, author = {Charlat, S and Ballard, JW and Merçot, H}, title = {What maintains noncytoplasmic incompatibility inducing Wolbachia in their hosts: a case study from a natural Drosophila yakuba population.}, journal = {Journal of evolutionary biology}, volume = {17}, number = {2}, pages = {322-330}, doi = {10.1046/j.1420-9101.2003.00676.x}, pmid = {15009266}, issn = {1010-061X}, mesh = {Analysis of Variance ; Animals ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; Cytoplasm/*microbiology ; Drosophila/*microbiology/*physiology ; Female ; Fertility/physiology ; Gabon ; Molecular Sequence Data ; Sequence Analysis, DNA ; Sex Ratio ; Sexual Behavior, Animal/*physiology ; Wolbachia/*genetics/physiology ; }, abstract = {Cytoplasmic incompatibility (CI) allows Wolbachia to invade hosts populations by specifically inducing sterility in crosses between infected males and uninfected females. In some species, non-CI inducing Wolbachia, that are thought to derive from CI-inducing ancestors, are common. In theory, the maintenance of such infections is not possible unless the bacterium is perfectly transmitted to offspring--and/or provides a fitness benefit to infected females. The present study aims to test this view by investigating a population of Drosophila yakuba from Gabon, West Africa. We did not find any evidence for CI using wild caught females. Infected females from the field transmitted the infection to 100% of their offspring. A positive effect on female fecundity was observed one generation after collecting, but this was not retrieved five generations later, using additional lines. Similarly, the presence of Wolbachia was found to affect mating behaviour, but the results of two experiments realized five generations apart were not consistent. Finally, Wolbachia was not found to affect sex ratio. Overall, our results would suggest that Wolbachia behaves like a neutral or nearly neutral trait in this species, and is maintained in the host by perfect maternal transmission.}, } @article {pmid14998511, year = {2004}, author = {Jolodar, A and Fischer, P and Büttner, DW and Brattig, NW}, title = {Wolbachia endosymbionts of Onchocerca volvulus express a putative periplasmic HtrA-type serine protease.}, journal = {Microbes and infection}, volume = {6}, number = {2}, pages = {141-149}, doi = {10.1016/j.micinf.2003.10.013}, pmid = {14998511}, issn = {1286-4579}, mesh = {Amino Acid Sequence ; Animals ; Molecular Sequence Data ; Onchocerca volvulus/*microbiology ; Periplasm/*enzymology ; Serine Endopeptidases/genetics/*metabolism ; Symbiosis/*physiology ; Wolbachia/*enzymology/genetics/physiology ; }, abstract = {Wolbachia are intracellular bacteria of many filarial nematodes. A mutualistic interaction between the endobacteria and the filarial host is likely, because the clearance of Wolbachia by tetracycline leads to the obstruction of embryogenesis and larval development. Databases were searched for exported molecules to identify candidates involved in this mutualism. Fragments of a Wolbachia serine protease from the human filarial parasite Onchocerca volvulus were obtained (Wol-Ov-HtrA) by the use of a PCR technique and primers based on the Rickettsia prowazekii genome. The deduced amino acid sequence exhibited 87% and 81% identity to the homologous Wolbachia proteases identified from Brugia malayi and Drosophila melanogaster, respectively. The full-length cDNA encodes 494 amino acids with a calculated mass of 54 kDa. Three characteristic features, (i) a catalytic triad of serine proteases, (ii) two PDZ domains and (iii) a putative signal peptide, classify the endobacterial protein as a member of the periplasmic HtrA family of proteases known to express chaperone and regulator activity of apoptosis. Using a rabbit antiserum raised against a recombinantly expressed 33-kDa fragment of Wol-Ov-HtrA, strong labelling of the antigen was found associated with endobacteria in hypodermis, oocytes, zygotes, all embryonic stages and microfilariae of O. volvulus. Staining of hypodermal cytoplasm surrounding the endobacteria indicated a possible release of the protein from the Wolbachia. The demonstration of Wol-Ov-HtrA-reactive IgG1 antibodies in sera of O. volvulus-infected persons indicated the exposure to the protein and its recognition by the human immune system. Wol-Ov-HtrA is a candidate for an exported Wolbachia protein that may interact with the filarial host metabolism.}, } @article {pmid14974684, year = {2003}, author = {Uesugi, R and Goka, K and Osakabe, M}, title = {Development of genetic differentiation and postzygotic isolation in experimental metapopulations of spider mites.}, journal = {Experimental & applied acarology}, volume = {31}, number = {3-4}, pages = {161-176}, pmid = {14974684}, issn = {0168-8162}, mesh = {Animals ; Female ; Genetic Variation ; Genetics, Population ; Insecticides ; Isoenzymes/genetics ; Malate Dehydrogenase/genetics ; Phosphoglucomutase/genetics ; Population Dynamics ; Selection, Genetic ; Tetranychidae/enzymology/*genetics/metabolism ; }, abstract = {We studied the development of genetic differentiation and postzygotic isolation in experimental metapopulations of the two-spotted spider mite, Tetranychus urticae Koch. A genetically diverse starter population was made by allowing six inbred sublines to interbreed. Then three migration patterns were tested: no migration, or one or three immigrants per subpopulation per generation. Variations in four traits were investigated: allozymes, acaricide resistance, diapause, and hatchability. In the allozymes, acaricide resistance, and diapause, genetic variation among subpopulations became high in metapopulations with no migration, but not in the others, which showed that one immigrant is enough to prevent genetic differentiation. Hatchability, which was decreased by interbreeding among the six sublines, gradually recovered in succeeding generations. In metapopulations with no migration, hatchability was reduced again after in-migration at the 15th generation. Different karyotypes or coadapted gene complexes can survive in different subpopulations by genetic drift, and both Wolbachia-infected and -noninfected subpopulations may be selected, which would lead to postzygotic isolation between isolated subpopulations. Our results indicate that sampling effects such as genetic drift or stochastic loss of Wolbachia produce postzygotic isolation in laboratory populations of spider mite.}, } @article {pmid14738900, year = {2004}, author = {Brattig, NW}, title = {Pathogenesis and host responses in human onchocerciasis: impact of Onchocerca filariae and Wolbachia endobacteria.}, journal = {Microbes and infection}, volume = {6}, number = {1}, pages = {113-128}, doi = {10.1016/j.micinf.2003.11.003}, pmid = {14738900}, issn = {1286-4579}, mesh = {Animals ; Humans ; Immunity, Cellular ; Immunity, Innate ; Inflammation/immunology/pathology ; Larva/physiology ; Onchocerca/*pathogenicity ; Onchocerciasis/immunology/*parasitology/pathology ; Symbiosis ; Virulence ; Wolbachia/*pathogenicity ; }, abstract = {Onchocerca volvulus is a tissue-invasive parasitic nematode causing skin and eye pathology in human onchocerciasis. The filariae habour abundant intracellular Wolbachia bacteria, now recognised as obligatory symbionts, and therefore emerging as a novel target for chemotherapy. Recent research demonstrates that both the filariae and endobacteria contribute to the pathogenesis of onchocerciasis, and molecules have been identified that promote inflammatory or counter-inflammatory immune mechanisms, divert the host's immune response or procure evasion of the parasite.}, } @article {pmid14733727, year = {2003}, author = {Hise, AG and Gillette-Ferguson, I and Pearlman, E}, title = {Immunopathogenesis of Onchocerca volvulus keratitis (river blindness): a novel role for TLR4 and endosymbiotic Wolbachia bacteria.}, journal = {Journal of endotoxin research}, volume = {9}, number = {6}, pages = {390-394}, doi = {10.1179/096805103225002746}, pmid = {14733727}, issn = {0968-0519}, support = {K08 AI054652/AI/NIAID NIH HHS/United States ; EY 10230/EY/NEI NIH HHS/United States ; EY 11373/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Keratitis/*etiology/immunology/microbiology/parasitology ; Membrane Glycoproteins/immunology/*metabolism ; Mice ; Mice, Inbred C3H ; Mice, Mutant Strains ; Models, Biological ; Onchocerca volvulus/*immunology/ultrastructure ; Onchocerciasis, Ocular/immunology/*microbiology/parasitology/pathology ; Receptors, Cell Surface/immunology/*metabolism ; Symbiosis/*immunology ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Wolbachia/*immunology/metabolism ; }, abstract = {Infection with the parasitic nematode Onchocerca volvulus is associated with inflammation of the skin and cornea that can lead to blindness. Corneal damage is thought to occur as a result of the host inflammatory responses to degenerating microfilariae in the eye. We have utilized a murine model of corneal inflammation (keratitis) to investigate the immune and inflammatory responses associated with river blindness. Soluble extracts of O. volvulus, a filarial species that contains the endosymbiont bacteria Wolbachia or Acanthocheilonema viteae (a nematode not naturally infected with the bacteria) were injected into mouse corneas. Inflammatory responses and corneal changes were measured. We demonstrated a major role for endosymbiont Wolbachia bacteria and Toll-like receptor 4 (TLR4) in the pathogenesis of ocular onchocerciasis.}, } @article {pmid14717905, year = {2004}, author = {Hughes, DP and Pamilo, P and Kathirithamby, J}, title = {Horizontal transmission of Wolbachia by strepsipteran endoparasites? A response to Noda et al., 2001.}, journal = {Molecular ecology}, volume = {13}, number = {2}, pages = {507-509}, doi = {10.1046/j.1365-294x.2003.02083.x}, pmid = {14717905}, issn = {0962-1083}, mesh = {Animals ; *Disease Transmission, Infectious ; Insecta/*microbiology/*parasitology/physiology ; Phylogeny ; Reproduction/physiology ; Wolbachia/*physiology ; }, } @article {pmid14711698, year = {2004}, author = {Benson, MJ and Gawronski, JD and Eveleigh, DE and Benson, DR}, title = {Intracellular symbionts and other bacteria associated with deer ticks (Ixodes scapularis) from Nantucket and Wellfleet, Cape Cod, Massachusetts.}, journal = {Applied and environmental microbiology}, volume = {70}, number = {1}, pages = {616-620}, pmid = {14711698}, issn = {0099-2240}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Cloning, Molecular ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Ixodes/cytology/*microbiology ; Massachusetts ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; }, abstract = {The diversity of bacteria associated with the deer tick (Ixodes scapularis) was assessed using PCR amplification, cloning, and sequencing of 16S rRNA genes originating from seven ticks collected from Nantucket Island and Wellfleet, Cape Cod, Mass. The majority of sequences obtained originated from gram-negative proteobacteria. Four intracellular bacteria were detected including strains of Ehrlichia, Rickettsia, and Wolbachia and an organism related to intracellular insect symbionts from the Cytophaga-Flavobacterium-Bacteroides group. Several strains of members of the Sphingomonadaceae were also detected in all but one tick. The results provide a view of the diversity of bacteria associated with I. scapularis ticks in the field.}, } @article {pmid14711652, year = {2004}, author = {Riegler, M and Charlat, S and Stauffer, C and Merçot, H}, title = {Wolbachia transfer from Rhagoletis cerasi to Drosophila simulans: investigating the outcomes of host-symbiont coevolution.}, journal = {Applied and environmental microbiology}, volume = {70}, number = {1}, pages = {273-279}, pmid = {14711652}, issn = {0099-2240}, mesh = {Animals ; *Biological Evolution ; Crosses, Genetic ; Drosophila/*microbiology/physiology ; Drosophila Proteins ; Female ; Fushi Tarazu Transcription Factors ; Genes, rRNA ; Homeodomain Proteins/genetics ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; Tephritidae/*microbiology/physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an endosymbiont of diverse arthropod lineages that can induce various alterations of host reproduction for its own benefice. Cytoplasmic incompatibility (CI) is the most common phenomenon, which results in embryonic lethality when males that bear Wolbachia are mated with females that do not. In the cherry fruit fly, Rhagoletis cerasi, Wolbachia seems to be responsible for previously reported patterns of incompatibility between populations. Here we report on the artificial transfer of two Wolbachia variants (wCer1 and wCer2) from R. cerasi into Drosophila simulans, which was performed with two major goals in mind: first, to isolate wCer1 from wCer2 in order to individually test their respective abilities to induce CI in the new host; and, second, to test the theoretical prediction that recent Wolbachia-host associations should be characterized by high levels of CI, fitness costs to the new host, and inefficient transmission from mothers to offspring. wCer1 was unable to develop in the new host, resulting in its rapid loss after successful injection, while wCer2 was established in the new host. Transmission rates of wCer2 were low, and the infection showed negative fitness effects, consistent with our prediction, but CI levels were unexpectedly lower in the new host. Based on these parameter estimates, neither wCer1 nor wCer2 could be naturally maintained in D. simulans. The experiment thus suggests that natural Wolbachia transfer between species might be restricted by many factors, should the ecological barriers be bypassed.}, } @article {pmid14707177, year = {2004}, author = {Pop, M and Kosack, DS and Salzberg, SL}, title = {Hierarchical scaffolding with Bambus.}, journal = {Genome research}, volume = {14}, number = {1}, pages = {149-159}, pmid = {14707177}, issn = {1088-9051}, support = {R01 LM006845/LM/NLM NIH HHS/United States ; R01-LM06845/LM/NLM NIH HHS/United States ; }, mesh = {Algorithms ; Animals ; Bacillus anthracis/genetics ; Benchmarking/methods ; Brucella/genetics ; Computational Biology ; Contig Mapping/*methods ; Databases, Genetic ; Dogs ; *Genome ; Genome, Bacterial ; Genomics/methods ; Haplotypes/genetics ; Shewanella/genetics ; *Software ; Software Validation ; Staphylococcus/genetics ; Wolbachia/genetics ; }, abstract = {The output of a genome assembler generally comprises a collection of contiguous DNA sequences (contigs) whose relative placement along the genome is not defined. A procedure called scaffolding is commonly used to order and orient these contigs using paired read information. This ordering of contigs is an essential step when finishing and analyzing the data from a whole-genome shotgun project. Most recent assemblers include a scaffolding module; however, users have little control over the scaffolding algorithm or the information produced. We thus developed a general-purpose scaffolder, called Bambus, which affords users significant flexibility in controlling the scaffolding parameters. Bambus was used recently to scaffold the low-coverage draft dog genome data. Most significantly, Bambus enables the use of linking data other than that inferred from mate-pair information. For example, the sequence of a completed genome can be used to guide the scaffolding of a related organism. We present several applications of Bambus: support for finishing, comparative genomics, analysis of the haplotype structure of genomes, and scaffolding of a mammalian genome at low coverage. Bambus is available as an open-source package from our Web site.}, } @article {pmid14706096, year = {2004}, author = {Hise, AG and Gillette-Ferguson, I and Pearlman, E}, title = {The role of endosymbiotic Wolbachia bacteria in filarial disease.}, journal = {Cellular microbiology}, volume = {6}, number = {2}, pages = {97-104}, doi = {10.1046/j.1462-5822.2003.00350.x}, pmid = {14706096}, issn = {1462-5814}, support = {K08 AI054652/AI/NIAID NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Brugia/immunology/*microbiology/pathogenicity ; Filariasis/*immunology/parasitology/*physiopathology ; Humans ; Onchocerca/immunology/*microbiology/pathogenicity ; *Symbiosis ; Wolbachia/growth & development/*immunology ; }, abstract = {In this review, we describe the pathogenic role of Wolbachia endosymbiotic bacteria in filarial diseases, focusing on the host innate immune responses to filarial and Wolbachia products. A description of the host pathogen recognition and early inflammatory responses including TLR4-mediated signalling, chemokine and cytokine responses and inflammatory cell recruitment is provided from human studies and from animal models of filarial disease. Finally, the impact of the discovery and characterization of Wolbachia on filarial research and treatment programmes is discussed.}, } @article {pmid14704183, year = {2003}, author = {Rasgon, JL and Scott, TW}, title = {Wolbachia and cytoplasmic incompatibility in the California Culex pipiens mosquito species complex: parameter estimates and infection dynamics in natural populations.}, journal = {Genetics}, volume = {165}, number = {4}, pages = {2029-2038}, pmid = {14704183}, issn = {0016-6731}, support = {GM-20092/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics/metabolism ; California ; Crosses, Genetic ; Culex/*microbiology/physiology ; Cytoplasm/*physiology ; Female ; *Genetics, Population ; *Infectious Disease Transmission, Vertical ; Infertility/microbiology ; Male ; Models, Genetic ; Polymerase Chain Reaction ; Rickettsiaceae Infections/*physiopathology/transmission ; Symbiosis/physiology ; Wolbachia/*physiology ; }, abstract = {Before maternally inherited bacterial symbionts like Wolbachia, which cause cytoplasmic incompatibility (CI; reduced hatch rate) when infected males mate with uninfected females, can be used in a program to control vector-borne diseases it is essential to understand their dynamics of infection in natural arthropod vector populations. Our study had four goals: (1) quantify the number of Wolbachia strains circulating in the California Culex pipiens species complex, (2) investigate Wolbachia infection frequencies and distribution in natural California populations, (3) estimate the parameters that govern Wolbachia spread among Cx. pipiens under laboratory and field conditions, and (4) use these values to estimate equilibrium levels and compare predicted infection prevalence levels to those observed in nature. Strain-specific PCR, wsp gene sequencing, and crossing experiments indicated that a single Wolbachia strain infects Californian Cx. pipiens. Infection frequency was near or at fixation in all populations sampled for 2 years along a >1000-km north-south transect. The combined statewide infection frequency was 99.4%. Incompatible crosses were 100% sterile under laboratory and field conditions. Sterility decreased negligibly with male age in the laboratory. Infection had no significant effect on female fecundity under laboratory or field conditions. Vertical transmission was >99% in the laboratory and approximately 98.6% in the field. Using field data, models predicted that Wolbachia will spread to fixation if infection exceeds an unstable equilibrium point above 1.4%. Our estimates accurately predicted infection frequencies in natural populations. If certain technical hurdles can be overcome, our data indicate that Wolbachia can invade vector populations as part of an applied transgenic strategy for vector-borne disease reduction.}, } @article {pmid14704179, year = {2003}, author = {Dean, MD and Ballard, KJ and Glass, A and Ballard, JW}, title = {Influence of two Wolbachia strains on population structure of East African Drosophila simulans.}, journal = {Genetics}, volume = {165}, number = {4}, pages = {1959-1969}, pmid = {14704179}, issn = {0016-6731}, mesh = {Africa, Eastern ; Animals ; Bacterial Outer Membrane Proteins ; Biological Evolution ; Cytoplasm/genetics ; DNA, Mitochondrial/genetics ; Demography ; Drosophila/genetics/*microbiology ; Female ; Genetic Variation ; *Genetics, Population ; Haplotypes ; Male ; Molecular Sequence Data ; Phylogeny ; Polymorphism, Genetic/*genetics ; Selection, Genetic ; Wolbachia/genetics/*physiology ; }, abstract = {Drosophila simulans is hypothesized to have originated in continental East Africa or Madagascar. In this study, we investigated evolutionary forces operating on mitochondrial DNA (mtDNA) in populations of D. simulans from Zimbabwe, Malawi, Tanzania, and Kenya. Variation in mtDNA may be affected by positive selection, background selection, demographic history, and/or any maternally inherited factor such as the bacterial symbiont Wolbachia. In East Africa, the wRi and wMa Wolbachia strains associate with the siII or siIII mitochondrial haplogroups, respectively. To ask how polymorphism relates to Wolbachia infection status, we sequenced 1776 bp of mitochondrial DNA and 1029 bp of the X-linked per locus from 79 lines. The two southern populations were infected with wRi and exhibited significantly reduced mtDNA variation, while Wolbachia-uninfected siII flies from Tanzania and Kenya showed high levels of mtDNA polymorphism. These are the first known populations of D. simulans that do not exhibit reduced mtDNA variation. We observed no mitochondrial variation in the siIII haplogroup regardless of Wolbachia infection status, suggesting positive or background selection. These populations offer a unique opportunity to monitor evolutionary dynamics in ancestral populations that harbor multiple strains of Wolbachia.}, } @article {pmid14702162, year = {2004}, author = {Lamb, TJ and Le Goff, L and Kurniawan, A and Guiliano, DB and Fenn, K and Blaxter, ML and Read, AF and Allen, JE}, title = {Most of the response elicited against Wolbachia surface protein in filarial nematode infection is due to the infective larval stage.}, journal = {The Journal of infectious diseases}, volume = {189}, number = {1}, pages = {120-127}, doi = {10.1086/380490}, pmid = {14702162}, issn = {0022-1899}, mesh = {Animals ; Antibodies, Bacterial/*blood ; Antibodies, Helminth/blood ; Bacterial Outer Membrane Proteins/*immunology ; Brugia malayi/immunology/microbiology/*physiology ; Disease Models, Animal ; Disease Progression ; Filariasis/*blood/diagnosis ; Filarioidea/microbiology/physiology ; Humans ; Larva/immunology ; Male ; Mice ; Mice, Inbred BALB C ; Species Specificity ; Symbiosis ; Wolbachia/*immunology ; }, abstract = {Immune responses to the intracellular Wolbachia bacteria of filarial nematodes are thought to contribute to the pathologic process of filarial infection. Here, we compare antibody responses of subjects living in an area where lymphatic filariasis is endemic with antibody responses elicited in a murine model of filarial infection, to provide evidence that the infective larval stage (L3), not adult nematodes, are the primary inducer of responses against Wolbachia. In human subjects, antibody responses to Brugia malayi Wolbachia surface protein (WSP) are most often correlated with antibody responses to the L3 stage of B. malayi. Analysis of anti-WSP responses induced in mice by different stages of the rodent filariae Litomosoides sigmodontis shows that the strongest anti-WSP response is elicited by the L3 stage. Although adult filarial nematode death may play a role in the generation of an anti-WSP response, it is the L3 stage that is the major source of immunogenic material, and incoming L3 provide a continual boosting of the anti-WSP response. Significant exposure to the endosymbiotic bacteria may occur earlier in nematode infection than previously thought, and the level of exposure to infective insect bites may be a key determinant of disease progression.}, } @article {pmid14686939, year = {2004}, author = {Czarnetzki, AB and Tebbe, CC}, title = {Detection and phylogenetic analysis of Wolbachia in Collembola.}, journal = {Environmental microbiology}, volume = {6}, number = {1}, pages = {35-44}, doi = {10.1046/j.1462-2920.2003.00537.x}, pmid = {14686939}, issn = {1462-2912}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Proteins/genetics ; Brain/microbiology ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/chemistry/isolation & purification ; Female ; Genes, rRNA ; In Situ Hybridization, Fluorescence ; Molecular Sequence Data ; Ovary/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Serotyping ; Wolbachia/*genetics/growth & development/*isolation & purification ; }, abstract = {Wolbachia are obligatory, cytoplasmatically inherited alpha-Proteobacteria which are known for infecting the reproductive tissues of many arthropods. Their prevalence in the large group of Collembola, however, is not known, except for PCR detection in the parthenogenetically reproducing species Folsomia candida (Order: Entomobryomorpha; Family: Isotomidae). In this study, fluorescence in situ hybridization on microscopic sections of F. candida specimens indicated that Wolbachia-related bacteria were restricted to tissues of the ovary and brain. PCR with primers designed to detect 16S rRNA genes of Wolbachia were positive with specimens from all of five geographically independent F. candida breeding stocks and with three parthenogenetic species from another order (Poduromorpha; Family Tullbergiidae), i.e. Mesaphorura italica, M. macrochaeta and Paratullbergia callipygos. In contrast, negative results were obtained with the two sexually reproducing species, Isotoma viridis (Isotomidae) and Protaphorura fimata (Poduromorpha; Onychiuridae). The ftsZ gene of Wolbachia could be PCR-amplified from all Wolbachia-positive hosts with the exception of M. macrochaeta. The phylogenetic distances of the ftsZ and 16S rRNA gene sequences reflected the phylogenetic distances of the host organisms but the sequences of Wolbachia were relatively closely related, indicating that Wolbachia infections took place after the Collembola had diversified. Our study confirms a monophyletic branch (supergroup E) of Collembola colonizing Wolbachia and indicates that this group is a sister group of supergroup A, the latter harbouring a high diversity of host organisms within the group of insects.}, } @article {pmid14663517, year = {2003}, author = {Kageyama, D and Ohno, S and Hoshizaki, S and Ishikawa, Y}, title = {Sexual mosaics induced by tetracycline treatment in the Wolbachia-infected adzuki bean borer, Ostrinia scapulalis.}, journal = {Genome}, volume = {46}, number = {6}, pages = {983-989}, doi = {10.1139/g03-082}, pmid = {14663517}, issn = {0831-2796}, mesh = {Animals ; Female ; Genitalia/drug effects/growth & development ; Larva/drug effects/growth & development/microbiology ; Male ; Moths/*drug effects/microbiology ; Sex Characteristics ; Sex Factors ; Tetracycline/*pharmacology ; Tibia/drug effects/growth & development ; Wings, Animal/drug effects/growth & development ; Wolbachia/*growth & development ; }, abstract = {Wolbachia-infected Ostrinia scapulalis (Lepidoptera: Crambidae) females generate all-female or nearly all-female broods. Curing the infection by tetracycline treatment during larval stages results in the generation of all-male broods in the next generation. Here we show that sexually mosaic offspring are produced by Wolbachia-infected females treated with tetracycline at the adult stage. The sexual mosaics had wings that were composed of distinctive female and male sectors. Besides wings, the sexually dimorphic mid tibiae displayed an intermediate morphology in some of the mosaics. Many of the mosaic individuals had an abnormal structure of the external genitalia as well, a combination of the male uncus and the female ovipositor. We assume that Wolbachia has a feminizing effect on O. scapulalis genetic males and, hence, incomplete curing of the Wolbachia infection results in the generation of sexually mosaic individuals.}, } @article {pmid14663516, year = {2003}, author = {Kageyama, D and Nishimura, G and Hoshizaki, S and Ishikawa, Y}, title = {Two kinds of sex ratio distorters in a moth, Ostrinia scapulalis.}, journal = {Genome}, volume = {46}, number = {6}, pages = {974-982}, doi = {10.1139/g03-083}, pmid = {14663516}, issn = {0831-2796}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/chemistry/genetics/isolation & purification ; Female ; Genotype ; Geography ; Male ; Molecular Sequence Data ; Moths/genetics/*microbiology ; Mutation ; Phenotype ; Sequence Analysis, DNA ; Sex Chromosome Aberrations ; *Sex Ratio ; Wolbachia/*genetics/growth & development ; }, abstract = {We surveyed female-biased sex ratio traits (SR traits) in a moth, Ostrinia scapulalis (Lepidoptera: Crambidae) from six geographic locations in Japan. A small proportion (8.3%) of females showed the SR trait. About half of them were associated with a Wolbachia infection (SRw+), whereas the remaining ones were not (SRw-). No significant differences in frequencies were detected among locations and years for either SRw+ or SRw- females. In SRw+ matrilines, the SR trait was inherited. Curing the Wolbachia infection by tetracycline at the larval stages resulted in all-male broods in the next generation. These characteristics were identical to the Wolbachia-induced SR trait observed previously in the related species Ostrinia furnacalis. The Wolbachia strain in O. scapulalis was indistinguishable from that in O. furnacalis in terms of the wsp and ftsZ gene sequences, suggesting either that infection originated from a common ancestor of the two hosts or that Wolbachia was transmitted horizontally between the host species. In SRw- matrilines, on the other hand, the SR trait was not due to bacterial infection but some other maternally inherited element, since antibiotic treatment had no effect and bacterial PCR did not give the characteristic product. In these matrilines, the SR trait occasionally failed to show, suggesting the presence of nuclear factor(s) suppressing the SR trait.}, } @article {pmid14662922, year = {2003}, author = {Fenollar, F and La Scola, B and Inokuma, H and Dumler, JS and Taylor, MJ and Raoult, D}, title = {Culture and phenotypic characterization of a Wolbachia pipientis isolate.}, journal = {Journal of clinical microbiology}, volume = {41}, number = {12}, pages = {5434-5441}, pmid = {14662922}, issn = {0095-1137}, mesh = {Animals ; Bacterial Proteins/isolation & purification ; Culicidae/*microbiology ; Electrophoresis, Polyacrylamide Gel ; Open Reading Frames/genetics ; Phenotype ; Phylogeny ; Polymerase Chain Reaction/methods ; Wolbachia/cytology/*genetics/growth & development/*isolation & purification ; }, abstract = {The recent isolation of Wolbachia pipientis in the continuous cell line Aa23, established from eggs of a strain of the Asian tiger mosquito Aedes albopictus, allowed us to perform extensive characterization of the isolate. Bacterial growth could be obtained in C6/36, another A. albopictus cell line, at 28 degrees C and in a human embryonic lung fibroblast monolayer at 28 and 37 degrees C, confirming that its host cell range is broader than was initially thought. The bacteria were best visualized by Diff-Quik and May-Grünwald-Giemsa staining. Proteins from 213 to 18 kDa with two major protein bands of 65 and 25 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By Western blotting with specific polyclonal mouse and rabbit antisera, dominant immunoreactive antigens were found at approximately 100, 80, and 30 kDa. The genome size was calculated to be 1,790 +/- 17 kb by pulsed-field gel electrophoresis. The sequence of the citrate synthase gene (gltA) of W. pipientis was determined by gene walking. Its position in the phylogenetic tree constructed with gltA confirmed that found in a phylogenetic tree constructed with 16S rRNA genes and that it belongs in the alpha subgroup of the class Proteobacteria and that it is closely related to but independent from the genera Ehrlichia, Anaplasma, and Neorickettsia.}, } @article {pmid14662876, year = {2003}, author = {Babu, S and Nutman, TB}, title = {Proinflammatory cytokines dominate the early immune response to filarial parasites.}, journal = {Journal of immunology (Baltimore, Md. : 1950)}, volume = {171}, number = {12}, pages = {6723-6732}, doi = {10.4049/jimmunol.171.12.6723}, pmid = {14662876}, issn = {0022-1767}, mesh = {Animals ; Antigen-Presenting Cells/immunology/metabolism/parasitology ; Antigens, Helminth/immunology ; Brugia malayi/growth & development/*immunology ; CD4-Positive T-Lymphocytes/cytology/immunology/metabolism/parasitology ; CD8-Positive T-Lymphocytes/cytology/immunology/metabolism/parasitology ; Cell Communication/immunology ; Cells, Cultured ; Cytokines/*biosynthesis ; Flow Cytometry ; Host-Parasite Interactions/immunology ; Humans ; Inflammation Mediators/*metabolism ; Lipopolysaccharides/pharmacology ; Lymphocyte Activation/immunology ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes/cytology/*immunology/*metabolism/parasitology ; Th1 Cells/cytology/immunology/metabolism/parasitology ; }, abstract = {Although the early human immune response to the infective-stage larvae (L3) of Brugia malayi has not been well-characterized in vivo (because of the inability to determine the precise time of infection), the consensus has been that it must involve a predominant Th2 environment. We have set up an in vitro system to study this early immune response by culturing PBMC from unexposed individuals with live L3 of B. malayi. After 24 h of culture, T cell responses were examined by flow cytometry and by quantitative real-time RT-PCR for multiple cytokines. T cells were activated early following exposure to L3 as indicated by up-regulation of surface markers CD69 and CD71. The frequency of T cells expressing proinflammatory Th1 cytokines (IFN-gamma, TNF-alpha, GM-CSF, IL-1alpha, and IL-8) but not Th2 cytokines (IL-4, IL-5, IL-6, IL-10, and IL-13) was significantly increased in response to L3. This T cell response occurred in both the CD4 and CD8 T cell compartment and was restricted to the effector/memory pool (CD45RO(+)). This T cell response was not due to LPS activity from the parasite or from its endosymbiont, Wolbachia; moreover, it required the presence of APC as well as direct contact with live L3. Real-time RT-PCR analysis of multiple cytokines in the T cells confirmed the increased expression of proinflammatory Th1 cytokines. Up-regulation of these cytokines suggests that the primary immune response to the live infective stage of the parasite is not predominantly Th2 in nature but rather dominated by a proinflammatory response.}, } @article {pmid14660690, year = {2004}, author = {Ballard, JW}, title = {Sequential evolution of a symbiont inferred from the host: Wolbachia and Drosophila simulans.}, journal = {Molecular biology and evolution}, volume = {21}, number = {3}, pages = {428-442}, doi = {10.1093/molbev/msh028}, pmid = {14660690}, issn = {0737-4038}, mesh = {Animals ; Base Sequence ; *Biological Evolution ; DNA, Mitochondrial/genetics ; Demography ; Drosophila/genetics/*microbiology ; Female ; Genetics, Population ; Geography ; Haplotypes ; Phylogeny ; Polymorphism, Genetic ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {This study aims to unravel the biogeography of a model symbiont/host system by exploiting the prediction that a symbiont will leave a signature of infection on the host. Specifically, a global sample of 1,442 Drosophila simulans from 33 countries and 64 sampling localities was employed to infer the phylogeography of the maternally inherited alpha-proteobacteria Wolbachia. Phylogenetic analyses, from three symbiont genes and 24 mtDNA genomes (excluding the A + T-rich region), showed that each of four Wolbachia strains infected D. simulans once. The global distribution and abundance of the Wolbachia strains and the three mtDNA haplogroups (D. simulans siI, siII and siIII) was then determined. Finally, network analyses of variable regions within siI (584 bp from seven additional lines) and siII (1,701 bp from 383 lines) facilitated a detailed biogeographic discussion. There is little variation in siIII and the haplogroup is restricted in its distribution. These data show how the history of an infection can be mapped by combining data from the symbiont and the host. They say little about the organismal history of the host because the mtDNA genome is a biased representation of the whole genome.}, } @article {pmid14651884, year = {2003}, author = {Komnenou, A and Egyed, Z and Sréter, T and Eberhard, ML}, title = {Canine onchocercosis in Greece: report of further 20 cases and molecular characterization of the parasite and its Wolbachia endosymbiont.}, journal = {Veterinary parasitology}, volume = {118}, number = {1-2}, pages = {151-155}, doi = {10.1016/j.vetpar.2003.09.007}, pmid = {14651884}, issn = {0304-4017}, mesh = {Animals ; DNA, Bacterial/*analysis ; Dog Diseases/microbiology/*parasitology/surgery ; Dogs ; Greece ; Onchocerca/isolation & purification/*microbiology ; Onchocerciasis, Ocular/microbiology/parasitology/surgery/*veterinary ; Phylogeny ; RNA, Ribosomal, 16S/analysis ; Symbiosis ; Wolbachia/classification/*isolation & purification ; }, abstract = {Recently, sporadic cases of subconjunctival Onchocerca infection have been reported in dogs in Greece and Hungary. Herein we report further cases from Greece and the results of the molecular analysis of Onchocerca sp. removed from Greek dogs and its Wolbachia endosymbionts. Twenty dogs of various breeds, 1-11 years of age with subconjunctival onchocercosis (4 cases each in right or left eye, 12 cases in both eyes) were presented having similar manifestations. Periorbital swelling, exophthalmos, lacrimation, discharge, photophobia, conjunctival congestion, corneal edema, protrusion of the nictitating membrane, and subconjunctival granuloma or cyst formation were the most important clinical signs. After surgical excision of the periocular masses containing the worms, all animals recovered fully from onchocercosis. Based on the similarities of the clinical picture of the Greek and Hungarian cases, the similar morphology of the Greek and Hungarian isolates, and the identical sequences of the cytochrome oxidase gene of the filarial parasites and that of the 16S ribosomal RNA gene from their Wolbachia endosymbionts, the Onchocerca sp. isolated from dogs in Greece and Hungary appears to belong to the same species.}, } @article {pmid14651648, year = {2003}, author = {Parvizi, P and Benlarbi, M and Ready, PD}, title = {Mitochondrial and Wolbachia markers for the sandfly Phlebotomus papatasi: little population differentiation between peridomestic sites and gerbil burrows in Isfahan province, Iran.}, journal = {Medical and veterinary entomology}, volume = {17}, number = {4}, pages = {351-362}, doi = {10.1111/j.1365-2915.2003.00451.x}, pmid = {14651648}, issn = {0269-283X}, mesh = {Animals ; Base Sequence ; Cytochromes b/genetics ; Disease Reservoirs/veterinary ; Female ; Gene Transfer Techniques ; *Gerbillinae/parasitology ; Haplotypes ; Host-Parasite Interactions ; Humans ; Insect Vectors/classification/*microbiology/parasitology ; Iran ; Leishmania major/pathogenicity/physiology ; Leishmaniasis, Cutaneous/*prevention & control/transmission ; Male ; Mitochondria/chemistry ; Phlebotomus/classification/*microbiology/parasitology ; Phylogeny ; Polymerase Chain Reaction/veterinary ; Population Dynamics ; Sequence Analysis, DNA ; Sex Factors ; Species Specificity ; Transgenes ; Wolbachia/*genetics/growth & development ; }, abstract = {In Iran, Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) is the main vector of Leishmania major Yakimoff & Schokhor (Kinetoplastida: Trypanosomatidae), the causative agent of rural zoonotic cutaneous leishmaniasis. This sandfly is abundant both in villages and in the burrows of the main reservoir host, the gerbil Rhombomys opimus (Licht.) (Rodentia: Gerbillidae). Populations of P. papatasi were sampled from the edges of villages in Isfahan province, using CDC miniature light traps in peridomestic sites and sticky papers placed at the entrances to gerbil burrows. Single peridomestic sites in two northern provinces were also sampled. Individual sandflies were characterized by PCR amplification and sequencing of fragments of their mitochondrial cytochrome b gene and of the wsp gene of endosymbiotic Wolbachia pipientis Hertig (alpha-Proteobacteria: Rickettsiaceae). The distributions of the haplotypes of these two maternally inherited genes were analysed to assess the population differentiation of P. papatasi, knowledge of which will be needed for planning control measures. For the first time these markers were used to characterize P. papatasi from gerbil burrows, and they indicated the absence not only of sympatric cryptic species but also of any long-term differentiation of lineages in different habitats. A single lineage of cytochrome b haplotypes was found, and both sexes in all populations had a high infection rate of the same A-group strain of Wolbachia (wPap). The distributions of cytochrome b haplotypes were consistent with females dispersing more than males, which has been reported for P. papatasi in other countries. The widespread distribution of wPap suggests that Wolbachia could be used to spread transgenes between populations of P. papatasi in different habitats.}, } @article {pmid14641977, year = {2003}, author = {Benlarbi, M and Ready, PD}, title = {Host-specific Wolbachia strains in widespread populations of Phlebotomus perniciosus and P. papatasi (Diptera: Psychodidae), and prospects for driving genes into these vectors of Leishmania.}, journal = {Bulletin of entomological research}, volume = {93}, number = {5}, pages = {383-391}, doi = {10.1079/ber2003251}, pmid = {14641977}, issn = {0007-4853}, mesh = {Animals ; Disease Transmission, Infectious/veterinary ; Female ; *Gene Transfer Techniques ; Genetic Vectors ; Host-Parasite Interactions ; Infectious Disease Transmission, Vertical/veterinary ; Insect Vectors/*microbiology/parasitology ; Leishmaniasis/prevention & control/*transmission ; Male ; Phlebotomus/*microbiology/parasitology ; Polymerase Chain Reaction/veterinary ; Reproduction ; Sex Factors ; Species Specificity ; Transgenes ; *Wolbachia/genetics/growth & development ; }, abstract = {A single strain of Wolbachia (alpha-proteobacteria, Rickettsiales) was found in widespread geographical populations of each of two Phlebotomus species, within which there was no indication of 'infectious speciation'. The two strains were identified by sequencing a fragment of wsp (a major surface protein gene), amplified by polymerase chain reaction from DNA extracted from the body parts of individual sandflies. Infection rates were high in the males and females of both sandflies, but they were lower for the B-group wPrn strain of Wolbachia in Phlebotomus perniciosus Newstead (60.3% overall) than for the A-group wPap strain in P. papatasi (Scopoli) (81.7%). Infections were frequent in the thorax, where Leishmania develops infective forms, as well as in the abdomen, where Wolbachia must infect the reproductive tissues to ensure its vertical transmission. These findings were related to knowledge of the population biology of Wolbachia in other insects, leading to the conclusion that this endosymbiont could be useful for driving transgenes through wild populations of both sandflies. This will require characterizing the cytoplasmic incompatibility phenotypes of Wolbachia-sandfly combinations, as well as estimating for them the incidence of paternal transmission and the fidelity of maternal transmission. Paternal transmission is one explanation for finding a single Wolbachia strain associated with all mitochondrial haplotypes and lineages of each sandfly species. However, this distribution pattern could also result from multiple horizontal transmissions or the failure of wsp to provide strain markers.}, } @article {pmid14645266, year = {2003}, author = {Sun, LV and Riegler, M and O'Neill, SL}, title = {Development of a physical and genetic map of the virulent Wolbachia strain wMelPop.}, journal = {Journal of bacteriology}, volume = {185}, number = {24}, pages = {7077-7084}, pmid = {14645266}, issn = {0021-9193}, mesh = {Blotting, Southern ; Chromosome Mapping ; *Genome, Bacterial ; Phenotype ; Virulence/genetics ; Wolbachia/*genetics/pathogenicity ; }, abstract = {We report here the construction of a physical and genetic map of the virulent Wolbachia strain, wMelPop. This map was determined by ordering 28 chromosome fragments that resulted from digestion with the restriction endonucleases FseI, ApaI, SmaI, and AscI and were resolved by pulsed-field gel electrophoresis. Southern hybridization was done with 53 Wolbachia-specific genes as probes in order to determine the relative positions of these restriction fragments and use them to serve as markers. Comparison of the resulting map with the whole genome sequence of the closely related benign Wolbachia strain, wMel, shows that the two genomes are largely conserved in gene organization with the exception of a single inversion in the chromosome.}, } @article {pmid14638788, year = {2003}, author = {Chirgwin, SR and Coleman, SU and Porthouse, KH and Nowling, JM and Punkosdy, GA and Klei, TR}, title = {Removal of Wolbachia from Brugia pahangi is closely linked to worm death and fecundity but does not result in altered lymphatic lesion formation in Mongolian gerbils (Meriones unguiculatus).}, journal = {Infection and immunity}, volume = {71}, number = {12}, pages = {6986-6994}, pmid = {14638788}, issn = {0019-9567}, support = {AI-19199-18/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; Antibodies, Bacterial/blood ; Brugia pahangi/drug effects/*microbiology/pathogenicity/*physiology ; Cytokines/metabolism ; DNA, Bacterial/analysis ; Female ; Fertility ; Filariasis/*drug therapy/parasitology/pathology/physiopathology ; Gerbillinae ; Kidney ; Lymph Nodes/immunology/pathology ; Male ; Polymerase Chain Reaction ; Tetracycline/*pharmacology/therapeutic use ; Wolbachia/*drug effects/genetics/isolation & purification ; }, abstract = {Approximately 30 years ago, researchers reported intracellular bacteria in filarial nematodes. These bacteria are relatives of the arthropod symbiont Wolbachia and occur in many filarial nematodes, including Brugia pahangi and Brugia malayi. Wolbachia bacteria have been implicated in a variety of roles, including filaria development and fecundity and the pathogenesis of lymphatic lesions associated with filarial infections. However, the role of the bacteria in worm biology or filarial disease is still not clear. The present experiments support previous data showing that tetracycline eliminates or reduces Wolbachia bacteria in B. pahangi in vivo. The elimination of Wolbachia was closely linked to a reduction in female fecundity and the viability of both sexes, suggesting that the killing of Wolbachia is detrimental to B. pahangi. The gerbils treated with tetracycline showed reduced levels of interleukin-4 (IL-4) and IL-5 mRNA in renal lymph nodes and spleens compared with the levels in B. pahangi-infected gerbils not treated with tetracycline. However, similar findings were noted in B. pahangi-infected gerbils treated with ivermectin, suggesting that the loss of circulating microfilariae, not the reduction of Wolbachia bacteria, was associated with the altered cytokine profile. Despite the change in T-cell cytokines, there was no difference in the sizes of renal lymph nodes isolated from gerbils in each treatment group. Furthermore, the numbers, sizes, or cellular compositions of granulomas examined in the lymphatics or renal lymph nodes did not differ with treatment. These data suggest that Wolbachia may not play a primary role in the formation of lymphatic lesions in gerbils chronically infected with B. pahangi.}, } @article {pmid14635812, year = {2003}, author = {Vala, F and Breeuwer, JA and Sabelis, MW}, title = {Sorting out the effects of Wolbachia, genotype and inbreeding on life-history traits of a spider mite.}, journal = {Experimental & applied acarology}, volume = {29}, number = {3-4}, pages = {253-264}, pmid = {14635812}, issn = {0168-8162}, mesh = {Animals ; Female ; Genetic Variation ; Genotype ; Inbreeding ; Male ; Sex Ratio ; Statistics, Nonparametric ; Symbiosis/physiology ; Tetranychidae/genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria manipulate host reproduction by inducing cytoplasmic incompatibility (CI) and sex ratio distortion. Wolbachia are transmitted from mother to offspring through the cytoplasm of the egg. Therefore, reproduction of Wolbachia is tightly coupled to reproduction of its host. Mathematical analysis predicts that in the course of evolution, traits that reduce the physiological costs of the infection will be selectively favored. For a Wolbachia-host system to evolve, traits under selection must have some genetic component and variation must be present in the population. We have previously established that highly inbred isofemale lines of the two-spotted spider mite Tetranychus urticae may differ regarding the effects of infection by Wolbachia, and that at least some of the traits affected had a genetic component. However, the effects measured could have been affected by the fact that the lines were severely inbred prior to the experiments. In this paper we attempt to distinguish between the effects of Wolbachia, isofemale line, and inbreeding. We show that Wolbachia did not affect longevity but infected females produced smaller clutch sizes, more daughter-biased sex ratios and had decreased F1 mortality; between-line variation was found for clutch size, F1 mortality and sex ratio; finally, inbreeding resulted in an overall reduction of clutch sizes, and a change in survival curves and mean longevity.}, } @article {pmid14629385, year = {2003}, author = {Tsutsui, ND and Kauppinen, SN and Oyafuso, AF and Grosberg, RK}, title = {The distribution and evolutionary history of Wolbachia infection in native and introduced populations of the invasive argentine ant (Linepithema humile).}, journal = {Molecular ecology}, volume = {12}, number = {11}, pages = {3057-3068}, doi = {10.1046/j.1365-294x.2003.01979.x}, pmid = {14629385}, issn = {0962-1083}, mesh = {Animals ; Ants/*microbiology ; Base Sequence ; Bayes Theorem ; DNA, Mitochondrial/genetics ; Electrophoresis, Agar Gel ; Genetic Markers/genetics ; *Genetics, Population ; *Geography ; Likelihood Functions ; Models, Genetic ; Molecular Sequence Data ; *Phylogeny ; Sequence Analysis, DNA ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia pipientis is a maternally transmitted bacterium that often alters the life history of its insect host to maximize transmission to subsequent generations. Here we report on the frequency and distribution of Wolbachia infection in a widespread invasive species, the Argentine ant (Linepithema humile). We screened 1175 individual Argentine ants from 89 nests on five continents and several islands, including numerous locations in both the native (South American) and introduced ranges. We detected Wolbachia in four of 11 native populations, but only one of 21 introduced populations was infected. In the Argentine ant's native range, the distribution of Wolbachia supergroups A and B was nonoverlapping. By coupling infection frequency data with behaviourally defined colony boundaries, we show that infected and uninfected colonies are often adjacent to one another, supporting the proposition that little female-mediated gene flow occurs among Argentine ant colonies. We also conduct a phylogenetic analysis, and show that the Wolbachia infecting both native and introduced populations of Argentine ants belong to two lineages that appear to be specialized on infecting New World ants. One other lineage of Wolbachia has undergone frequent, recent episodes of horizontal transmission between distantly related, introduced insect hosts.}, } @article {pmid14629360, year = {2003}, author = {Mouton, L and Henri, H and Bouletreau, M and Vavre, F}, title = {Strain-specific regulation of intracellular Wolbachia density in multiply infected insects.}, journal = {Molecular ecology}, volume = {12}, number = {12}, pages = {3459-3465}, doi = {10.1046/j.1365-294x.2003.02015.x}, pmid = {14629360}, issn = {0962-1083}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; DNA Primers ; Electrophoresis, Agar Gel ; *Genetic Variation ; Population Density ; Species Specificity ; *Symbiosis ; Wasps/*microbiology ; Wolbachia/*genetics/physiology ; }, abstract = {Vertically transmitted symbionts suffer a severe reduction in numbers when they pass through host generations, resulting in genetic homogeneity or even clonality of their populations. Wolbachia endosymbionts that induce cytoplasmic incompatibility in their hosts depart from this rule, because cytoplasmic incompatibility actively maintains multiple infection within hosts. Hosts and symbionts are thus probably under peculiar selective pressures that must shape the way intracellular bacterial populations are regulated. We studied the density and location of Wolbachia within adult Leptopilina heterotoma, a haplodiploid wasp that is parasitic on Drosophila and that is naturally infected with three Wolbachia strains, but for which we also obtained one simply infected and two doubly infected lines. Comparison of these four lines by quantitative polymerase chain reaction using a real-time detection system showed that total Wolbachia density varies according to the infection status of individuals, while the specific density of each Wolbachia strain remains constant regardless of the presence of other strains. This suggests that Wolbachia strains do not compete with one another within the same host individual, and that a strain-specific regulatory mechanism is operating. We discuss the regulatory mechanisms that are involved, and how this process might have evolved as a response to selective pressures acting on both partners.}, } @article {pmid14629013, year = {2003}, author = {Thipaksorn, A and Jamnongluk, W and Kittayapong, P}, title = {Molecular evidence of Wolbachia infection in natural populations of tropical odonates.}, journal = {Current microbiology}, volume = {47}, number = {4}, pages = {314-318}, doi = {10.1007/s00284-002-4010-4}, pmid = {14629013}, issn = {0343-8651}, mesh = {Animals ; Bacterial Outer Membrane Proteins/analysis/*genetics ; Bacterial Proteins/analysis/genetics ; DNA, Bacterial/chemistry/isolation & purification ; Genes, Bacterial ; Insecta/*microbiology ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology ; Thailand ; Wolbachia/classification/*genetics/*isolation & purification ; }, abstract = {Wolbachia are endosymbiotic bacteria that cause reproductive alterations in numerous arthropod species. Using a PCR-based method, we found that, out of 33 odonate species, four species were infected with Wolbachia. This finding represents the first record of Wolbachia infection in tropical odonates. Identical wsp gene sequences were found in the Wolbachia-infected common odonate species, Agriocnemis f. femina, collected from different locations in Thailand. The infection frequencies in several natural populations suggest that replacement of uninfected populations by Wolbachia-infected ones has recently occurred in this damselfly species.}, } @article {pmid14615218, year = {2003}, author = {Jeyaprakash, A and Hoy, MA and Allsopp, MH}, title = {Bacterial diversity in worker adults of Apis mellifera capensis and Apis mellifera scutellata (Insecta: Hymenoptera) assessed using 16S rRNA sequences.}, journal = {Journal of invertebrate pathology}, volume = {84}, number = {2}, pages = {96-103}, doi = {10.1016/j.jip.2003.08.007}, pmid = {14615218}, issn = {0022-2011}, mesh = {Animals ; Bacteria/*genetics/isolation & purification ; Base Sequence ; Bees/*microbiology ; DNA, Bacterial/*analysis ; *Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S/*analysis ; Restriction Mapping ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology ; }, abstract = {High-fidelity PCR of 16S rRNA sequences was used to identify bacteria associated with worker adults of the honeybee subspecies Apis mellifera capensis and Apis mellifera scutellata. An expected approximately 1.5-kb DNA band, representing almost the entire length of the 16S rRNA gene, was amplified from both subspecies and cloned. Ten unique sequences were obtained: one sequence each clustered with Bifidobacterium (Gram-positive eubacteria), Lactobacillus (Gram-positive eubacteria), and Gluconacetobacter (Gram-negative alpha-proteobacteria); two sequences each clustered with Simonsiella (beta-proteobacteria) and Serratia (gamma-proteobacteria); and three sequences each clustered with Bartonella (alpha-proteobacteria). Although the sequences relating to these six bacterial genera initially were obtained from either A. m. capensis or A. m. scutellata or both, newly designed honeybee-specific 16S rRNA primers subsequently amplified all sequences from all individual workers of both subspecies. Attempts to amplify these sequences from eggs have failed. However, the wsp primers designed to amplify Wolbachia DNA from arthropods, including these bees, consistently produced a 0.6-kb DNA band from individual eggs, indicating that amplifiable bacterial DNA was present. Hence, the 10 bacteria could have been acquired orally from workers or from other substrates. This screening of 16S rRNA sequences from A. m. capensis and A. m. scutellata found sequences related to Lactobacillus and Bifidobacterium which previously had been identified from other honeybee subspecies, as well as sequences related to Bartonella, Gluconacetobacter, Simonsiella/Neisseria, and Serratia, which have not been identified previously from honeybees.}, } @article {pmid14597281, year = {2003}, author = {Bazzocchi, C and Genchi, C and Paltrinieri, S and Lecchi, C and Mortarino, M and Bandi, C}, title = {Immunological role of the endosymbionts of Dirofilaria immitis: the Wolbachia surface protein activates canine neutrophils with production of IL-8.}, journal = {Veterinary parasitology}, volume = {117}, number = {1-2}, pages = {73-83}, doi = {10.1016/j.vetpar.2003.07.013}, pmid = {14597281}, issn = {0304-4017}, mesh = {Animals ; Bacterial Outer Membrane Proteins/immunology ; Chemotaxis/immunology ; Dirofilaria/*microbiology ; Dirofilaria immitis/microbiology ; Dirofilariasis/parasitology ; Dog Diseases/parasitology ; Dogs ; Immunoglobulin G/biosynthesis ; Interleukin-8/biosynthesis ; Neutrophils/metabolism ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; *Symbiosis ; Wolbachia/*immunology ; }, abstract = {Filarial nematodes, including Dirofilaria immitis and D. repens, harbour intracellular bacteria belonging to the genus Wolbachia. These bacteria have been implicated in the pathogenesis of filarial diseases, possibly through their endotoxins. Recent studies have shown that a major surface protein of Wolbachia (WSP) induces a specific IgG response in hosts infected by D. immitis. WSP from the Wolbachia of D. immitis was produced in recombinant form. The purified protein was used in stimulation assays on canine neutrophils. The assays performed using a modified Boyden chamber showed that WSP stimulates neutrophil chemokinesis. In addition, RT-PCR revealed increased production of chemokine IL-8 by cells incubated with this protein. Neutrophils have been shown to play a major role in the pathogenesis of river blindness, and to accumulate in the nodules of onchocerciasis patients. In dogs infected by D. immitis, neutrophils accumulate in kidneys and in the wall of pulmonary arteries. As shown by our studies, Wolbachia could contribute to these inflammatory phenomena through its surface protein WSP, independently from its endotoxin component.}, } @article {pmid14580056, year = {2003}, author = {Vala, F and Breeuwer, JA and Sabelis, MW}, title = {No variation for Wolbachia-induced hybrid breakdown in two populations of a spider mite.}, journal = {Experimental & applied acarology}, volume = {29}, number = {1-2}, pages = {1-12}, pmid = {14580056}, issn = {0168-8162}, mesh = {Animals ; Crosses, Genetic ; Female ; Male ; Reproduction/*physiology ; Sex Ratio ; Tetranychidae/*microbiology/physiology ; Wolbachia/*growth & development ; }, abstract = {Wolbachia are cytoplasmically transmitted bacteria that infect several species of mites. In the two-spotted spider mite Tetranychus urticae Koch this symbiont can induce reproductive incompatibility. Wolbachia-induced reproductive incompatibility is observed in crosses between Wolbachia-infected (W) males and uninfected (U) females. This incompatibility is expressed in F1 broods as male-biased sex ratios, an effect called cytoplasmic incompatibility (CI). However, in the two-spotted spider mite, Wolbachia-induced reproductive incompatibility may extend to the F2: broods of virgin F1 females from U x W crosses sometimes suffer increased mortality rates. This F2 effect is called hybrid breakdown (HB). Several isofemale lines derived from mites collected from rose and cucumber plants had been previously tested for CI. Here we report on the results obtained for HB.}, } @article {pmid14561283, year = {2003}, author = {Hunter, MS and Perlman, SJ and Kelly, SE}, title = {A bacterial symbiont in the Bacteroidetes induces cytoplasmic incompatibility in the parasitoid wasp Encarsia pergandiella.}, journal = {Proceedings. Biological sciences}, volume = {270}, number = {1529}, pages = {2185-2190}, pmid = {14561283}, issn = {0962-8452}, mesh = {Animals ; Bacteroidaceae/*physiology ; Crosses, Genetic ; Cytoplasm/*physiology ; Fertility/physiology ; Parthenogenesis/physiology ; Sequence Analysis, DNA ; Sex Ratio ; Symbiosis/*physiology ; Wasps/*microbiology ; }, abstract = {Vertically transmitted symbionts of arthropods have been implicated in several reproductive manipulations of their hosts. These include cytoplasmic incompatibility (CI), parthenogenesis induction in haplodiploid species (PI), feminization and male killing. One symbiont lineage in the alpha-Proteobacteria, Wolbachia, is the only bacterium known to cause all of these effects, and has been thought to be unique in causing CI, in which the fecundity of uninfected females is reduced after mating with infected males. Here, we provide evidence that an undescribed symbiont in the Bacteroidetes group causes CI in a sexual population of the parasitic wasp Encarsia pergandiella. Wasps were crossed in all four possible combinations of infected and uninfected individuals. In the cross predicted to be incompatible, infected (I) males x uninfected (U) females, progeny production was severely reduced, with these females producing only 12.6% of the number of progeny in other crosses. The incompatibility observed in this haplodiploid species was the female mortality type; dissections showed that most progeny from the incompatible cross died as eggs. The 16S rDNA sequence of this symbiont is 99% identical to a parthenogenesis-inducing symbiont in other Encarsia, and 96% identical to a feminizing symbiont in haplodiploid Brevipalpus mites. Thus, this recently discovered symbiont lineage is capable of inducing three of the four principal manipulations of host reproduction known to be caused by Wolbachia.}, } @article {pmid14532065, year = {2003}, author = {Kikuchi, Y and Fukatsu, T}, title = {Diversity of Wolbachia endosymbionts in heteropteran bugs.}, journal = {Applied and environmental microbiology}, volume = {69}, number = {10}, pages = {6082-6090}, pmid = {14532065}, issn = {0099-2240}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Cytoskeletal Proteins/genetics ; DNA, Bacterial/analysis ; *Genetic Variation ; Heteroptera/classification/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA ; *Symbiosis ; Wolbachia/*classification/genetics ; }, abstract = {An extensive survey of Wolbachia endosymbionts in Japanese terrestrial heteropteran bugs was performed by PCR detection with universal primers for wsp and ftsZ genes of Wolbachia, cloning of the PCR products, restriction fragment length polymorphism analysis of infecting Wolbachia types, and molecular phylogenetic characterization of all the detected Wolbachia strains. Of 134 heteropteran species from 19 families examined, Wolbachia infection was detected in 47 species from 13 families. From the 47 species, 59 Wolbachia strains were identified. Of the 59 strains, 16 and 43 were assigned to A group and B group in the Wolbachia phylogeny, respectively. The 47 species of Wolbachia-infected bugs were classified into 8 species with A infection, 28 species with B infection, 2 species with AA infection, 3 species with AB infection, 5 species with BB infection, and 1 species with ABB infection. Molecular phylogenetic analysis showed little congruence between Wolbachia phylogeny and host systematics, suggesting frequent horizontal transfers of Wolbachia in the evolutionary course of the Heteroptera. The phylogenetic analysis also revealed several novel lineages of Wolbachia. Based on statistical analyses of the multiple infections, we propose a hypothetical view that, in the heteropteran bugs, interactions between coinfecting Wolbachia strains are generally not intense and that Wolbachia coinfections have been established through a stochastic process probably depending on occasional horizontal transfers.}, } @article {pmid14524541, year = {2003}, author = {Mahilum, MM and Storch, V and Becker, N}, title = {Molecular and electron microscopic identification of Wolbachia in Culex pipiens complex populations from the Upper Rhine Valley, Germany, and Cebu City, Philippines.}, journal = {Journal of the American Mosquito Control Association}, volume = {19}, number = {3}, pages = {206-210}, pmid = {14524541}, issn = {8756-971X}, mesh = {Animals ; Culex/*microbiology ; DNA, Bacterial/analysis ; Female ; Germany ; Male ; Microscopy, Electron ; Philippines ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; Wolbachia/*genetics/isolation & purification/*ultrastructure ; }, abstract = {We detected Wolbachia infections in Culex pipiens complex mosquitoes from the Upper Rhine Valley, Germany, and Cebu City, Philippines, with the use of polymerase chain reaction (PCR) amplification of the 16S rDNA of the bacteria and further confirmation by electron microscopy. Species of 5 different autogenous and anautogenous populations of Culex pipiens from Germany and 1 population of Culex quinquefasciatus from the Philippines were used in this study. Larvae were sampled from different localities and reared to the adult stage for further processing. Whole adult males and the heads of adult females were processed by PCR, whereas the abdomens of females were used for electron microscopic study when an infection was detected by PCR. All Culex populations assayed by PCR showed infection of Wolbachia at rates between 10 and 100%. Females from different populations exhibited higher infection rates than did the males, and, likewise, Cx. pipiens populations that were autogenous showed higher infection rates of Wolbachia than did anautogenous individuals. In Cx. quinquefasciatus populations, males were infected at 33.33% and females, only 10%. We studied the ultrastructure of Wolbachia in the ovaries of species belonging to Cx. pipiens complex by means of transmission electron microscopy. The bacteria exhibited typical morphology for Wolbachia with 3 enveloping membranes.}, } @article {pmid14512953, year = {2003}, author = {Keeling, MJ and Jiggins, FM and Read, JM}, title = {The invasion and coexistence of competing Wolbachia strains.}, journal = {Heredity}, volume = {91}, number = {4}, pages = {382-388}, doi = {10.1038/sj.hdy.6800343}, pmid = {14512953}, issn = {0018-067X}, mesh = {Models, Biological ; Species Specificity ; Wolbachia/classification/*physiology ; }, abstract = {Cytoplasmic incompatibility between arthropods infected with different strains of Wolbachia has been proposed as an important mechanism for speciation. However, a basic requirement for this mechanism is the coexistence of different strains in neighbouring populations. Here we test whether this required coexistence is possible in a spatial context. Continuous-time models for the behaviour of one and two strains of Wolbachia within a single well-mixed population demonstrate the Allee effect and founder control, such that one strain is always driven extinct. In contrast, discretised spatial models show patchy persistence of the two strains although coexistence within the same habitat is rare. A simplified model of such founder control suggests that it is fragmentation of (or barriers within) the habitat rather than space itself that leads to persistence.}, } @article {pmid14511562, year = {2003}, author = {Kassem, HA and Hassan, AN and Abdel-Hamid, I and Osman, G and El Khalab, EM and Madkour, MA}, title = {Wolbachia infection and the expression of cytoplasmic incompatibility in sandflies (Diptera: Psychodidae) from Egypt.}, journal = {Annals of tropical medicine and parasitology}, volume = {97}, number = {6}, pages = {639-644}, doi = {10.1179/000349803225001391}, pmid = {14511562}, issn = {0003-4983}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; DNA, Bacterial/analysis ; Egypt ; Female ; Insect Vectors ; Male ; Parasitology/methods ; Phlebotomus/*microbiology/physiology ; Polymerase Chain Reaction/methods ; Reproduction ; Rickettsiaceae Infections/*diagnosis/drug therapy ; Species Specificity ; Tetracyclines ; *Wolbachia/genetics ; }, abstract = {A PCR-based method was used to screen four laboratory colonies of sandflies for Wolbachia infection. The colonies - one of Phlebotomus langeroni, one of P. bergeroti and two of P. papatasi - were all derived from sandflies collected in Egypt. Only one of the colonies, derived from P. papatasi collected in Sinai, was found infected. The sequence of the PCR product for this colony was identical to that previously reported for the Wolbachia in P. papatasi from Israel. The induction with tetracycline of cytoplasmic incompatibility (CI) in flies from the P. papatasi (Sinai) colony was then investigated, through reciprocal crosses between treated and untreated P. papatasi siblings. Partial CI expression was attained in the crosses involving antibiotic-treated (i.e. uninfected) females, whether the males used were infected with Wolbachia or had also been cleared of Wolbachia by antibiotic treatment. Most (75%) of the eggs oviposited by uninfected females that had been crossed with infected males, and most (58%) of those laid by uninfected females that had been crossed with uninfected males, failed to hatch. These results provide the first published evidence showing that Wolbachia infection in sandflies is advantageous to the insects. The failure to detect Wolbachia in one of the colonies derived from Egyptian P. papatasi or in the colonies derived from Egyptian P. bergeroti and P. langeroni may indicate that the inter- and intra-specific spread of Wolbachia is discontinuous, even within one country.}, } @article {pmid14506854, year = {2003}, author = {Nirgianaki, A and Banks, GK and Frohlich, DR and Veneti, Z and Braig, HR and Miller, TA and Bedford, ID and Markham, PG and Savakis, C and Bourtzis, K}, title = {Wolbachia infections of the whitefly Bemisia tabaci.}, journal = {Current microbiology}, volume = {47}, number = {2}, pages = {93-101}, doi = {10.1007/s00284-002-3969-1}, pmid = {14506854}, issn = {0343-8651}, mesh = {Animals ; Aphids/*microbiology ; Bacterial Outer Membrane Proteins/genetics ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Hemiptera/*microbiology ; Insecta/microbiology ; Lepidoptera/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {We report the first systematic survey for the presence of Wolbachia endosymbionts in aphids and whiteflies, particularly different populations and biotypes of Bemisia tabaci. Additional agriculturally important species included were predator species, leafhoppers, and lepidopterans. We used a polymerase chain reaction (PCR)-based detection assay with ribosomal 16S rDNA and Wolbachia cell surface protein (wsp) gene primers. Wolbachia were detected in a number of whitefly populations and species, whitefly predators, and one leafhopper species; however, none of the aphid species tested were found infected. Single, double, and triple infections were detected in some of the B. tabaci populations. PCR and phylogenetic analysis of wsp gene sequences indicated that all Wolbachia strains found belong to group B. Topologies of the optimal tree derived by maximum likelihood (ML) and a ML tree in which Wolbachia sequences from B. tabaci are constrained to be monophyletic are significantly different. Our results indicate that there have been at least four independent Wolbachia infection events in B. tabaci. The importance of the presence of Wolbachia infections in B. tabaci is discussed.}, } @article {pmid14501992, year = {2003}, author = {Hoerauf, A}, title = {Control of filarial infections: not the beginning of the end, but more research is needed.}, journal = {Current opinion in infectious diseases}, volume = {16}, number = {5}, pages = {403-410}, doi = {10.1097/00001432-200310000-00006}, pmid = {14501992}, issn = {0951-7375}, mesh = {Clinical Trials as Topic ; Filariasis/*prevention & control ; Filaricides/therapeutic use ; Humans ; Mosquito Control ; Onchocerciasis/prevention & control ; Vaccines ; }, abstract = {PURPOSE OF REVIEW: Infections with the filarial nematodes affect more than 150 million people mainly in the tropics. The very successful efforts to control filarial infections, however, have to be sustained by new tools that require long-term commitment to research. This review, focusing on reports from 2002 and 2003, highlights recent advances in research on immunology, understanding of pathogenesis and drug development in lymphatic filariasis and onchocerciasis research with potential relevance to the generation of new tools for control.

RECENT FINDINGS: Dramatic improvement has been achieved in the control of lymphatic filariasis and onchocerciasis by vector control and mass treatment with microfilaricidal drugs. Additional tools that could help in regional elimination or, ultimately, eradication of filariasis may arise from the development of new drugs or a vaccine. Research into the immune responses mediating protection or pathology has provided new insights into the pathways that lead to effector function and immunosuppression, such as T regulatory responses, as well as into genetic predispositions from the host's side, and to the identification of vaccine candidates that show protection in animal models. Recognition of the role the Wolbachia endosymbionts may play in activating the innate immune system has altered our understanding of immunopathology of filariasis and adverse reactions to microfilaricidal drugs. Wolbachia spp. have also proven to be suitable targets for the development of a long-term sterilizing or potentially macrofilaricidal drug.

SUMMARY: This review summarizes recent developments in the control of filariasis, in particular lymphatic filariasis and onchocerciasis, as well as in modern research into the immunity of filariasis and new drug development that could lead to additional tools necessary for sustained success in filariasis control.}, } @article {pmid13678709, year = {2003}, author = {Brownstein, JS and Hett, E and O'Neill, SL}, title = {The potential of virulent Wolbachia to modulate disease transmission by insects.}, journal = {Journal of invertebrate pathology}, volume = {84}, number = {1}, pages = {24-29}, doi = {10.1016/s0022-2011(03)00082-x}, pmid = {13678709}, issn = {0022-2011}, support = {AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Age Factors ; Animals ; Culicidae/*parasitology ; Cytoplasm ; Dengue/*transmission ; Fertility ; Host-Parasite Interactions ; Insect Vectors/parasitology ; *Models, Biological ; Pest Control, Biological/*methods ; Wolbachia/*pathogenicity/physiology ; }, abstract = {A virulent strain of Wolbachia has recently been identified in Drosophila that drastically reduces adult lifespan. It has been proposed that this phenotype might be introduced into insect disease vector populations to reduce pathogen transmission. Here we model the requirements for spread of such an agent and the associated reduction in disease transmission. First, a simulation of mosquito population age structure was used to describe the age distribution of mosquitoes transmitting dengue virus. Second, given varying levels of cytoplasmic incompatibility and fecundity effect, the maximum possible longevity reduction that would allow Wolbachia to invade was obtained. Finally, the two models were combined to estimate the reduction in disease transmission according to different introduction frequencies. With strong CI and limited effect of fecundity, an introduction of Wolbachia with an initial frequency of 0.4 could result in a 60-80% reduction of transmitting mosquitoes. Greater reductions are possible at higher initial release rates.}, } @article {pmid13678706, year = {2003}, author = {Ikeda, T and Ishikawa, H and Sasaki, T}, title = {Infection density of Wolbachia and level of cytoplasmic incompatibility in the Mediterranean flour moth, Ephestia kuehniella.}, journal = {Journal of invertebrate pathology}, volume = {84}, number = {1}, pages = {1-5}, doi = {10.1016/s0022-2011(03)00106-x}, pmid = {13678706}, issn = {0022-2011}, mesh = {Animals ; Chaperonin 60/genetics ; Cytoplasm/parasitology ; DNA Primers ; Female ; Host-Parasite Interactions/*physiology ; Male ; Moths/*genetics/*parasitology ; Phylogeny ; Polymerase Chain Reaction ; Rickettsiaceae Infections/genetics ; Testis/microbiology ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia, a causative agent of various reproductive changes in arthropods, induces cytoplasmic incompatibility (CI) in the Mediterranean flour moth, Ephestia kuehniella. Two strains of E. kuehniella, Yokohama and Tsuchiura, harbor closely related Wolbachia, but the Yokohama strain expresses stronger CI than the Tsuchiura strain. A transinfected E. kuehniella strain that harbors the Wolbachia derived from the almond moth Cadra cautella, expresses weak CI at a similar level to the Tsuchiura strain. In the present study, we measured the Wolbachia density in the testis of the three E. kuehniella strains in order to examine the effects of bacterial strain and infection load on the expression of CI. When individuals of the same strain were compared, a correlation of bacterial density to CI level was observed. In addition, the Wolbachia density was higher in the Yokohama strain than the Tsuchiura strain in agreement with the CI levels expressed. However, this relationship did not hold in the comparison between the naturally infected and transinfected strains that carried phylogenetically distant Wolbachia.}, } @article {pmid12965021, year = {2003}, author = {Moreau, J and Rigaud, T}, title = {Variable male potential rate of reproduction: high male mating capacity as an adaptation to parasite-induced excess of females?.}, journal = {Proceedings. Biological sciences}, volume = {270}, number = {1523}, pages = {1535-1540}, pmid = {12965021}, issn = {0962-8452}, mesh = {Animals ; *Birth Rate ; Female ; Feminization/microbiology ; Fertility/physiology ; Isopoda/microbiology/*physiology ; Male ; Population Dynamics ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Numerous animals are known to harbour intracytoplasmic symbionts that gain transmission to a new host generation via female eggs and not male sperm. Bacteria of the genus Wolbachia are a typical example. They infect a large range of arthropod species and manipulate host reproduction in several ways. In terrestrial isopods (woodlice), Wolbachia are responsible for converting males into females (feminization (F)) in some species, or for infertility in certain host crosses in other species (cytoplasmic incompatibility (CI)). Wolbachia with the F phenotype impose a strong excess of females on their host populations, while Wolbachia expressing CI do not. Here, we test the possibility that male mating capacity (MC) is correlated with Wolbachia-induced phenotype. We show that males of isopod hosts harbouring F Wolbachia possess a strong MC (i.e. are able to mate with several females in a short time), while those of species harbouring CI Wolbachia possess a weaker MC. This pattern may be explained either by the selection of high MC following the increase in female-biased sex ratios, or because the F phenotype would lead to population extinction in species where MC is not sufficiently high. This last hypotheses is nevertheless more constrained by population structure.}, } @article {pmid12964989, year = {2003}, author = {Weeks, AR and Velten, R and Stouthamer, R}, title = {Incidence of a new sex-ratio-distorting endosymbiotic bacterium among arthropods.}, journal = {Proceedings. Biological sciences}, volume = {270}, number = {1526}, pages = {1857-1865}, pmid = {12964989}, issn = {0962-8452}, mesh = {Animals ; Arthropods/*physiology ; Base Sequence ; Cytophaga/*genetics/*physiology ; DNA Primers ; DNA, Ribosomal/genetics ; Likelihood Functions ; Molecular Sequence Data ; Phylogeny ; *Sex Ratio ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {Many intracellular micro-organisms are now known to cause reproductive abnormalities and other phenomena in their hosts. The endosymbiont Wolbachia is the best known of these reproductive manipulators owing to its extremely high incidence among arthropods and the diverse host effects it has been implicated as causing. However, recent evidence suggests that another intracellular bacterium, a Cytophaga-like organism (CLO), may also induce several reproductive effects in its hosts. Here, we present the first survey of arthropod hosts for infection by the CLO. We use a sensitive hemi-nested polymerase chain reaction method to screen 223 species from 20 arthropod orders for infection by the CLO and Wolbachia. The results indicate that, although not as prevalent as Wolbachia, the CLO infects a significant number of arthropod hosts (ca. 7.2%). In addition, double infections of the CLO and Wolbachia were found in individuals of seven arthropod species. Sequencing analysis of the 16S rDNA region of the CLO indicates evidence for horizontal transmission of the CLO strains. We discuss these results with reference to future studies on host effects induced by intracellular micro-organisms.}, } @article {pmid19262760, year = {2003}, author = {Bordenstein, SR and Fitch, DH and Werren, JH}, title = {Absence of wolbachia in nonfilariid nematodes.}, journal = {Journal of nematology}, volume = {35}, number = {3}, pages = {266-270}, pmid = {19262760}, issn = {0022-300X}, abstract = {Intracellular bacteria of the genus Wolbachia are among the most abundant endosymbionts on the planet, occurring in at least two major phyla-the Arthropoda and Nematoda. Current surveys of Wolbachia distribution have found contrasting patterns within these groups. Whereas Wolbachia are widespread and occur in all three major subphyla of arthropods, with estimates placing them in at least several million arthropod species, the presence of nematodes carrying Wolbachia is currently confined to the filariids, in which they occur at appreciable frequencies. It has been hypothesized that Wolbachia entered the ancestor of modern-day filariids in a single acquisition event, and subsequently cospeciated with their filariid hosts. To further investigate this hypothesis, we examined the broader distribution of Wolbachia in nematodes using a polymerase chain reaction (PCR) assay in a diverse set of nonfilariid species. The assay consisted of three different types of PCR screens on adults of 20 secernentean nematode species (14 rhabditids, 2 strongylid parasites of vertebrates; 1 diplogasterid; 3 cephalobid relatives, 1 myolaim, and 1 filariid) and two adenophorean species (plectids). Two PCR screens were specific to the 16S rDNA and ftsZ protein coding gene of Wolbachia; and the third screen was specific to the 18S rDNA of the nematodes. Based upon our results, we conclude that Wolbachia are absent in all 21 non-filariid species encompassing all the major groups of the Secernentea and two species of Adenophorea, from which the Secernentea derived. The absence of Wolbachia in these non-filariids is consistent with the hypothesis that Wolbachia entered the nematode phylum once, in an ancestral lineage of extant filariids.}, } @article {pmid12943116, year = {2003}, author = {Armbruster, P and Damsky, WE and Giordano, R and Birungi, J and Munstermann, LE and Conn, JE}, title = {Infection of New- and Old-World Aedes albopictus (Diptera: Culicidae) by the intracellular parasite Wolbachia: implications for host mitochondrial DNA evolution.}, journal = {Journal of medical entomology}, volume = {40}, number = {3}, pages = {356-360}, doi = {10.1603/0022-2585-40.3.356}, pmid = {12943116}, issn = {0022-2585}, support = {F32AI09969-02/AI/NIAID NIH HHS/United States ; R01AI54139/AI/NIAID NIH HHS/United States ; R29AI40116/AI/NIAID NIH HHS/United States ; AI44793/AI/NIAID NIH HHS/United States ; R01 AI054139/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*classification/genetics/*microbiology ; Africa ; Animals ; Asia ; DNA, Mitochondrial/genetics/*isolation & purification ; Female ; Geography ; North America ; South America ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia are cytoplasmically inherited, endosymbiotic bacteria known to infect a wide variety of arthropods. Polymerase chain reaction (PCR) amplification of the Wolbachia surface protein (wsp) gene was used to assay the infection of geographically disparate populations of Aedes albopictus (Skuse) by Wolbachia. Nine North American, four South American, one Hawaiian, and four Old World populations of A. albopictus were all doubly infected with both the wAlbA and wAlbB strains of Wolbachia. A 365-bp region of the wAlbA wsp gene was sequenced from seven geographically disparate host populations, and all sequences were identical. Similarly, a 474-bp region of the wAlbB wsp gene was sequenced from the same populations, and all sequences were identical. These results suggest a role for Wolbachia infection in causing the previously established pattern of low mitochondrial DNA variability, but average nuclear gene diversity, within and among populations of A. albopictus.}, } @article {pmid12941392, year = {2003}, author = {Tram, U and Ferree, PM and Sullivan, W}, title = {Identification of Wolbachia--host interacting factors through cytological analysis.}, journal = {Microbes and infection}, volume = {5}, number = {11}, pages = {999-1011}, doi = {10.1016/s1286-4579(03)00192-8}, pmid = {12941392}, issn = {1286-4579}, mesh = {Animals ; Cell Cycle Proteins/physiology ; Centrosome/physiology ; Cytoskeleton/physiology ; Drosophila melanogaster/cytology/*microbiology ; Host-Parasite Interactions/physiology ; Microtubules/physiology ; Reproduction/physiology ; Wasps/cytology/growth & development/*microbiology ; Wolbachia/*physiology ; }, abstract = {Manipulation of host reproduction and efficient maternal transmission have facilitated the global spread of Wolbachia through millions of insect species. Cytological studies of the most common Wolbachia-induced phenotype, cytoplasmic incompatibility (CI), demonstrate that Wolbachia induce CI by altering host cell cycle timing. Cytological analyses also suggest that microtubules and motor proteins may play a role in the maternal and somatic transmission of Wolbachia.}, } @article {pmid12939620, year = {2003}, author = {Gotoh, T and Noda, H and Hong, XY}, title = {Wolbachia distribution and cytoplasmic incompatibility based on a survey of 42 spider mite species (Acari: Tetranychidae) in Japan.}, journal = {Heredity}, volume = {91}, number = {3}, pages = {208-216}, doi = {10.1038/sj.hdy.6800329}, pmid = {12939620}, issn = {0018-067X}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Crosses, Genetic ; Cytoplasm/*microbiology ; Female ; Insecta ; Japan ; Male ; Phylogeny ; Reproduction ; Sex Ratio ; Species Specificity ; Survival Rate ; Symbiosis ; Tetranychidae/*microbiology ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Wolbachia are a group of maternally inherited bacteria that infect a wide range of arthropods. Wolbachia infections are known to result in the expression of various abnormal reproductive phenotypes, the best known being cytoplasmic incompatibility. The first systematic survey of 42 spider mite species in Japan revealed that seven species (16.7%) were infected with Wolbachia. Wolbachia in the spider mites were grouped into three subgroups in supergroup B by phylogenetic analyses of the wsp gene. Most spider mites did not show cytoplasmic incompatibility when infected males were crossed with uninfected females. However, all infected populations of Panonychus mori and Oligonychus gotohi (five and four populations, respectively) possessed modification-positive strains of Wolbachia, and the cytoplasmic incompatibility decreased egg hatchability and female ratio of the spider mites. Thus, some Wolbachia strains cause sex ratio distortion in their hosts.}, } @article {pmid12933853, year = {2003}, author = {Punkosdy, GA and Addiss, DG and Lammie, PJ}, title = {Characterization of antibody responses to Wolbachia surface protein in humans with lymphatic filariasis.}, journal = {Infection and immunity}, volume = {71}, number = {9}, pages = {5104-5114}, pmid = {12933853}, issn = {0019-9567}, mesh = {Adolescent ; Adult ; Aged ; Amino Acid Sequence ; Animals ; Antibodies, Bacterial/*blood ; Antigens, Bacterial/genetics ; Bacterial Proteins/genetics/*immunology ; Brugia malayi/microbiology ; Case-Control Studies ; Elephantiasis, Filarial/*immunology/*microbiology/parasitology ; Epitope Mapping ; Female ; Haiti ; Humans ; Immunoglobulin G/blood ; Male ; Membrane Proteins/genetics/immunology ; Middle Aged ; Symbiosis ; Testicular Hydrocele/immunology/microbiology/parasitology ; Time Factors ; Wolbachia/genetics/*immunology/pathogenicity ; }, abstract = {Symbiotic Wolbachia organisms of filarial nematodes have received much attention as possible chemotherapy targets and disease-causing organisms. In order to further investigate the association between anti-Wolbachia immune responses and chronic filarial disease in humans, antibody responses to Wolbachia surface protein (WSP) were assayed in serum samples collected from 232 individuals living in Leogane, Haiti, an area where Wuchereria bancrofti infection is endemic, and from 67 North Americans with no history of lymphatic filariasis. As opposed to antifilarial antibody responses, which were largely influenced by the patient's infection status, the prevalence and levels of anti-WSP immunoglobulin G (IgG) antibodies among individuals with lymphedema or hydrocele were significantly greater than those in gender- and infection-matched individuals without disease. In at least one case, the anti-WSP IgG response was coincident with the onset of lymphedema development, and among anti-WSP-positive women with lymphedema, anti-WSP IgG levels were negatively correlated with the duration of lymphedema. The presence of anti-WSP IgG was also associated with the severity of inguinal adenopathy among men with hydrocele. In addition to the presence of anti-WSP antibodies among Haitians, 15 of 67 (22%) serum samples collected from individuals from North America, where filariasis is not endemic, were also positive for anti-WSP antibodies. In comparison to those from Haitians, anti-WSP antibodies from North Americans primarily recognized a distinct region of WSP located within the highly conserved second transmembrane domain. The results of this study demonstrate that anti-WSP antibody responses are associated with the presence of chronic filarial morbidity and not filarial infection status in humans and suggest that WSP should be further studied as a potential trigger for the development of filarial disease.}, } @article {pmid12897857, year = {2003}, author = {Harris, HL and Braig, HR}, title = {Sperm chromatin remodelling and Wolbachia-induced cytoplasmic incompatibility in Drosophila.}, journal = {Biochemistry and cell biology = Biochimie et biologie cellulaire}, volume = {81}, number = {3}, pages = {229-240}, doi = {10.1139/o03-053}, pmid = {12897857}, issn = {0829-8211}, mesh = {Animals ; Chromatin/*physiology ; Cytoplasm/physiology ; DNA Methylation ; Drosophila/genetics/microbiology/*physiology ; Female ; Fertility/genetics/physiology ; Male ; Mutation ; Spermatogenesis/physiology ; Spermatozoa/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis is an obligate bacterial endosymbiont, which has successfully invaded approximately 20% of all insect species by manipulating their normal developmental patterns. Wolbachia-induced phenotypes include parthenogenesis, male killing, and, most notably, cytoplasmic incompatibility. In the future these phenotypes might be useful in controlling or modifying insect populations but this will depend on our understanding of the basic molecular processes underlying insect fertilization and development. Wolbachia-infected Drosophila simulans express high levels of cytoplasmic incompatibility in which the sperm nucleus is modified and does not form a normal male pronucleus when fertilizing eggs from uninfected females. The sperm modification is somehow rescued in eggs infected with the same strain of Wolbachia. Thus, D. simulans has become an excellent model organism for investigating the manner in which endosymbionts can alter reproductive programs in insect hosts. This paper reviews the current knowledge of Drosophila early development and particularly sperm function. Developmental mutations in Drosophila that are known to affect sperm function will also be discussed.incompatibility.}, } @article {pmid12881548, year = {2003}, author = {Pennisi, E}, title = {Evolution 2003 meeting. Genetic deficiency blamed on bacteria.}, journal = {Science (New York, N.Y.)}, volume = {301}, number = {5632}, pages = {459}, doi = {10.1126/science.301.5632.459}, pmid = {12881548}, issn = {1095-9203}, mesh = {Animals ; *Bacterial Physiological Phenomena ; *Biological Evolution ; Digestion ; Diploidy ; Female ; Haploidy ; Insecta/*genetics/*microbiology ; Male ; Ovum/microbiology ; *Ploidies ; Sex Characteristics ; *Symbiosis ; Wolbachia/physiology ; }, } @article {pmid12871912, year = {2003}, author = {Reynolds, KT and Thomson, LJ and Hoffmann, AA}, title = {The effects of host age, host nuclear background and temperature on phenotypic effects of the virulent Wolbachia strain popcorn in Drosophila melanogaster.}, journal = {Genetics}, volume = {164}, number = {3}, pages = {1027-1034}, pmid = {12871912}, issn = {0016-6731}, mesh = {*Age Factors ; Animals ; Crosses, Genetic ; Drosophila melanogaster/*genetics/*microbiology ; Female ; Fertility/physiology ; Longevity ; Male ; Population Dynamics ; Species Specificity ; *Temperature ; Wolbachia/*pathogenicity ; }, abstract = {Because of their obligate endosymbiotic nature, Wolbachia strains by necessity are defined by their phenotypic effects upon their host. Nevertheless, studies on the influence of host background and environmental conditions upon the manifestation of Wolbachia effects are relatively uncommon. Here we examine the behavior of the overreplicating Wolbachia strain popcorn in four different Drosophila melanogaster backgrounds at two temperatures. Unlike other strains of Wolbachia in Drosophila, popcorn has a major fitness impact upon its hosts. The rapid proliferation of popcorn causes cells to rupture, resulting in the premature death of adult hosts. Apart from this effect, we found that popcorn delayed development time, and host background influenced both this trait and the rate of mortality associated with infection. Temperature influenced the impact of popcorn upon host mortality, with no reduction in life span occurring in flies reared at 19 degrees. No effect upon fecundity was found. Contrary to earlier reports, popcorn induced high levels of incompatibility when young males were used in tests, and CI levels declined rapidly with male age. The population dynamics of popcorn-type infections will therefore depend on environmental temperature, host background, and the age structure of the population.}, } @article {pmid12866789, year = {2003}, author = {Hartmann, N and Stuckas, H and Lucius, R and Bleiss, W and Theuring, F and Kalinna, BH}, title = {Trans-species transfer of Wolbachia: microinjection of Wolbachia from litomosoides sigmodontis into Acanthocheilonema viteae.}, journal = {Parasitology}, volume = {126}, number = {Pt 6}, pages = {503-511}, pmid = {12866789}, issn = {0031-1820}, mesh = {Animals ; DNA, Bacterial/analysis ; DNA, Helminth/analysis ; Dipetalonema/genetics/*microbiology/ultrastructure ; Filarioidea/*microbiology/ultrastructure ; Gerbillinae ; Microinjections ; Ornithodoros ; Polymerase Chain Reaction ; Symbiosis ; Wolbachia/genetics/*physiology/ultrastructure ; }, abstract = {Intracellular bacteria of the genus Wolbachia are found in most filarial nematodes, but are lacking in some species like Acanthocheilonema viteae. Due to their symbiotic nature and their role in the pathology of filarial infections they are considered to be potential targets for intervention against filarial infections in man. Infection of A. viteae (a species which does not naturally carry Wolbachia) with Wolbachia bacteria could allow comparative studies on the effect of the endobacterium on the parasite and on the host's immune systems. As a step towards such studies we microinjected adult female A. viteae with Wolbachia obtained from Litomosoides sigmodontis. The bacteria were isolated from L. sigmodontis by density-gradient centrifugation, microinjected into A. viteae worms and bacterial DNA detected by PCR with Wolbachia specific primers (ftsZ gene). Microinjected worms were cultured in vitro, and 81% survived for 10 days. Implantation of microinjected worms into Meriones unguiculatus, the rodent host of A. viteae resulted in 38% survival. The DNA of the microinjected worms recovered from jirds 8 weeks after implantation contained Wolbachia DNA as shown by PCR, suggesting that Wolbachia of L. sigmodontis can be horizontally transmitted to A. viteae.}, } @article {pmid12860708, year = {2003}, author = {Unver, A and Rikihisa, Y and Kawahara, M and Yamamoto, S}, title = {Analysis of 16S rRNA gene sequences of Ehrlichia canis, Anaplasma platys, and Wolbachia species from canine blood in Japan.}, journal = {Annals of the New York Academy of Sciences}, volume = {990}, number = {}, pages = {692-698}, doi = {10.1111/j.1749-6632.2003.tb07445.x}, pmid = {12860708}, issn = {0077-8923}, support = {R01AI47407/AI/NIAID NIH HHS/United States ; }, mesh = {Anaplasma/classification/*genetics ; Animals ; Base Sequence ; DNA, Ribosomal/*genetics ; Dogs/blood/*microbiology ; Ehrlichia canis/classification/*genetics ; Japan ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Sequence Alignment ; Sequence Homology, Nucleic Acid ; Wolbachia/classification/*genetics ; }, abstract = {In the present study, three canine blood samples from Japan, that were suspected to be ehrlichia positive were examined. After sequencing the 16S rRNA genes, each dog was found to be infected either with Ehrlichia canis (Kagoshima 1), Anaplasma platys (Okinawa 1), or Wolbachia sp. (Okinawa 2). Phylogenic analysis was performed on these sequences. The nearly entire 16S rRNA sequence of Kagoshima 1 was found to be most similar to the sequences from Oklahoma and Venezuela E. canis strains (1 base pair difference out of 1,387, 99.9% sequence identity). The 16S rRNA gene sequence of Okinawa 1 showed the closest DNA identity to the French strain of A. platys (1 base deletion out of 1,385 bp, 99.6% sequence identity). The 16S rRNA gene sequence of Okinawa 2 illustrated the closest DNA identity to that of a Wolbachia sp. from Dirofilaria immitis (98.9% sequence similarity). These data imply a low diversity within E. canis strains and within A. platys strains, including those strains reported in this study. This is also the first demonstration of Wolbachia DNA in dog blood, suggesting the involvement of Wolbachia sp. in canine febrile illnesses.}, } @article {pmid12860672, year = {2003}, author = {Taylor, MJ}, title = {Wolbachia in the inflammatory pathogenesis of human filariasis.}, journal = {Annals of the New York Academy of Sciences}, volume = {990}, number = {}, pages = {444-449}, doi = {10.1111/j.1749-6632.2003.tb07409.x}, pmid = {12860672}, issn = {0077-8923}, mesh = {Elephantiasis, Filarial/pathology/physiopathology ; Humans ; Inflammation ; Onchocerciasis/pathology/physiopathology ; Phylogeny ; Rickettsiaceae Infections/pathology/*physiopathology ; *Wolbachia/classification/isolation & purification ; }, abstract = {Filarial nematodes cause some of the most debilitating diseases in tropical medicine. Recent studies, however, have implicated the parasites' endosymbiotic Wolbachia bacteria, rather than the nematode, as the cause of inflammatory-mediated filarial disease. Soluble extracts of a variety of filarial species stimulate innate inflammatory responses, which are absent or reduced when using extracts derived from species either devoid of bacteria, or those cleared of bacteria by antibiotics. Characterization of the molecular nature of the bacterial derived inflammatory stimulus points toward an endotoxin-like activity that is dependent on the pattern recognition receptors CD14 and TLR4 and can be inhibited by lipid A antagonists. TLR4 dependent inflammation has been shown to occur in the systemic inflammatory adverse reaction to Brugia malayi following anti-filarial chemotherapy and in the development of neutrophil-mediated ocular inflammation in a mouse model of river blindness. The development of acute and severe inflammatory responses in people infected with Brugia malayi and Onchocerca volvulus is associated with the release of Wolbachia into the blood following death or damage of the worms after anti-filarial chemotherapy. Together these studies suggest that Wolbachia are the principal cause of acute inflammatory filarial disease. Accumulated exposure to acute episodes of inflammation may also underlie the development of chronic filarial pathology. The use of antibiotic therapy to target Wolbachia of filarial parasites may therefore provide a means to prevent the development of filarial pathology.}, } @article {pmid12856318, year = {2002}, author = {Fischer, P and Schmetz, C and Bandi, C and Bonow, I and Mand, S and Fischer, K and Büttner, DW}, title = {Tunga penetrans: molecular identification of Wolbachia endobacteria and their recognition by antibodies against proteins of endobacteria from filarial parasites.}, journal = {Experimental parasitology}, volume = {102}, number = {3-4}, pages = {201-211}, doi = {10.1016/s0014-4894(03)00058-4}, pmid = {12856318}, issn = {0014-4894}, mesh = {Amino Acid Sequence ; Animals ; Aspartate Aminotransferases/chemistry/genetics/*immunology ; Bacterial Outer Membrane Proteins/chemistry/genetics/*immunology ; Bacterial Proteins/chemistry/genetics/*immunology ; Base Sequence ; Child ; *Cytoskeletal Proteins ; DNA Primers/chemistry ; DNA, Bacterial/chemistry ; DNA, Ribosomal/chemistry ; Female ; Humans ; Immune Sera/immunology ; Immunohistochemistry ; Microscopy, Electron ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Sequence Alignment ; Siphonaptera/*microbiology/ultrastructure ; Wolbachia/classification/genetics/immunology/*isolation & purification ; }, abstract = {In search of Wolbachia in human parasites, Wolbachia were identified in the sand flea Tunga penetrans. PCR and DNA sequencing of the bacterial 16S rDNA, the ftsZ cell division protein, the Wolbachia surface protein (wsp) and the Wolbachia aspartate aminotransferase genes revealed a high similarity to the respective sequences of endosymbionts of filarial nematodes. Using these sequences a phylogenetic tree was generated, that indicates a close relationship between Wolbachia from T. penetrans and from filarial parasites, but possibly as a member of a new supergroup. Ultrastructural studies showed that Wolbachia are abundant in the ovaries of neosomic fleas, whereas other, smaller and morphologically distinct, bacteria were observed in the lumen of the intestine. Wolbachia were labeled by immunohistology and immunogold electron microscopy using polyclonal antibodies against wsp of Drosophila, of the filarial parasite Dirofilaria immitis, or against hsp 60 from Yersinia enterocolitica. These results show that as in filariasis, humans with tungiasis are exposed to Wolbachia. Furthermore, antisera raised against proteins of Wolbachia from arthropods or from filarial parasites can be immunologically cross-reactive.}, } @article {pmid12854423, year = {2003}, author = {Vinogradova, EB and Fedorova, MV and Shaikevich, EV and Zakharov, IA}, title = {Endosymbiotic bacterium Wolbachia pipientis in synanthropic populations of the mosquito Culex pipiens pipiens L. (Diptera, Culicidae).}, journal = {Doklady biological sciences : proceedings of the Academy of Sciences of the USSR, Biological sciences sections}, volume = {389}, number = {}, pages = {172-175}, pmid = {12854423}, issn = {0012-4966}, mesh = {Animals ; Base Sequence ; Culex/*microbiology ; DNA/chemistry ; Drosophila/genetics ; Electron Transport Complex IV/genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Homology, Nucleic Acid ; Wolbachia/*pathogenicity ; }, } @article {pmid12816546, year = {2003}, author = {McGarry, HF and Pfarr, K and Egerton, G and Hoerauf, A and Akue, JP and Enyong, P and Wanji, S and Kläger, SL and Bianco, AE and Beeching, NJ and Taylor, MJ}, title = {Evidence against Wolbachia symbiosis in Loa loa.}, journal = {Filaria journal}, volume = {2}, number = {1}, pages = {9}, pmid = {12816546}, issn = {1475-2883}, abstract = {BACKGROUND: The majority of filarial nematode species are host to Wolbachia bacterial endosymbionts, although a few including Acanthocheilonema viteae, Onchocerca flexuosa and Setaria equina have been shown to be free of infection. Comparisons of species with and without symbionts can provide important information on the role of Wolbachia symbiosis in the biology of the nematode hosts and the contribution of the bacteria to the development of disease. Previous studies by electron microscopy and PCR have failed to detect intracellular bacterial infection in Loa loa. Here we use molecular and immunohistological techniques to confirm this finding. METHODS: We have used a combination of PCR amplification of bacterial genes (16S ribosomal DNA [rDNA], ftsZ and Wolbachia surface protein [WSP]) on samples of L. loa adults, third-stage larvae (L3) and microfilariae (mf) and immunohistology on L. loa adults and mf derived from human volunteers to determine the presence or absence of Wolbachia endosymbionts. Samples used in the PCR analysis included 5 adult female worms, 4 adult male worms, 5 mf samples and 2 samples of L3. The quality and purity of nematode DNA was tested by PCR amplification of nematode 5S rDNA and with diagnostic primers from the target species and used to confirm the absence of contamination from Onchocerca sp., Mansonella perstans, M. streptocerca and Wuchereria bancrofti. Immunohistology was carried out by light and electron microscopy on L. loa adults and mf and sections were probed with rabbit antibodies raised to recombinant Brugia malayi Wolbachia WSP. Samples from nematodes known to be infected with Wolbachia (O. volvulus, O. ochengi, Litomosoides sigmodontis and B. malayi) were used as positive controls and A. viteae as a negative control. RESULTS: Single PCR analysis using primer sets for the bacterial genes 16S rDNA, ftsZ, and WSP were negative for all DNA samples from L. loa. Positive PCR reactions were obtained from DNA samples derived from species known to be infected with Wolbachia, which confirmed the suitability of the primers and PCR conditions. The quality and purity of nematode DNA samples was verified by PCR amplification of 5S rDNA and with nematode diagnostic primers. Additional analysis by 'long PCR' failed to produce any further evidence for Wolbachia symbiosis. Immunohistology of L. loa adults and mf confirmed the results of the PCR with no evidence for Wolbachia symbiosis. CONCLUSION: DNA analysis and immunohistology provided no evidence for Wolbachia symbiosis in L. loa.}, } @article {pmid12810042, year = {2003}, author = {Chirgwin, SR and Nowling, JM and Coleman, SU and Klei, TR}, title = {Brugia pahangi and Wolbachia: the kinetics of bacteria elimination, worm viability, and host responses following tetracycline treatment.}, journal = {Experimental parasitology}, volume = {103}, number = {1-2}, pages = {16-26}, doi = {10.1016/s0014-4894(03)00063-8}, pmid = {12810042}, issn = {0014-4894}, support = {AI-19199-18/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; Blotting, Southern ; Brugia pahangi/drug effects/*microbiology/physiology ; Cell Count ; Cytokines/analysis ; DNA, Bacterial/analysis ; Female ; Fertility ; Filariasis/*drug therapy/immunology/parasitology ; Gerbillinae ; Male ; Peritoneal Cavity/pathology ; Polymerase Chain Reaction ; Random Allocation ; Tetracycline/*pharmacology/therapeutic use ; Wolbachia/*drug effects/isolation & purification/physiology ; }, abstract = {Wolbachia spp., first reported from filariae nearly 30 years ago, have been suggested to contribute to the pathogenesis associated with human filarial infection. Tetracycline has been used to cure filariae of Wolbachia, as a novel means of chemotherapeutic treatment for both ocular and lymphatic filariasis. Tetracycline treatment of L4 or adult Brugia pahangi in vivo resulted in Wolbachia clearance. Less tetracycline was required to clear Wolbachia when treatment began at the L4 stage, compared with adults. Female worms died earlier than male worms when tetracycline was administered at the L4 stage. In all cases, Wolbachia clearance was closely associated with worm death. Worm recoveries decreased following the L4-L5 molt, suggesting tetracycline does not interrupt molting in this model system. Despite worm death and the assumed release of both bacterial- and worm-derived molecules, differences in inflammatory cell population and T cell cytokine mRNA profiles were negligible between tetracycline-treated and non-treated B. pahangi infected gerbils. These data suggest the contribution of Wolbachia to the in vivo induction of the gerbil immune response to B. pahangi may be small.}, } @article {pmid12807775, year = {2003}, author = {Veneti, Z and Clark, ME and Zabalou, S and Karr, TL and Savakis, C and Bourtzis, K}, title = {Cytoplasmic incompatibility and sperm cyst infection in different Drosophila-Wolbachia associations.}, journal = {Genetics}, volume = {164}, number = {2}, pages = {545-552}, pmid = {12807775}, issn = {0016-6731}, mesh = {Animals ; Crosses, Genetic ; Cysts/microbiology ; Cytoplasm/metabolism ; Drosophila melanogaster/*microbiology ; Female ; Genotype ; Male ; Microscopy, Confocal ; Phenotype ; Spermatogenesis ; Spermatozoa/metabolism ; Temperature ; Testis/microbiology ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia are a group of maternally transmitted obligatory intracellular alpha-proteobacteria that infect a wide range of arthropod and nematode species. Wolbachia infection in Drosophila in most cases is associated with the induction of cytoplasmic incompatibility (CI), manifested as embryonic lethality of offspring in a cross between infected males and uninfected females. While the molecular basis of CI is still unknown, it has been suggested that two bacterial functions are involved: mod (for modification) modifies the sperm during spermatogenesis and resc (for rescue) acts in the female germline and/or in early embryos, neutralizing the modification. There is considerable variation in the level of incompatibility in different Wolbachia/host interactions. We examine the relationship between the levels of CI in a number of naturally infected and transinfected Drosophila hosts and the percentage of Wolbachia-infected sperm cysts. Our results indicate the presence of two main groups of Drosophila-Wolbachia associations: group I, which exhibits a positive correlation between CI levels and the percentage of infected sperm cysts (mod(+) phenotype), and group II, which does not express CI (mod(-) phenotype) irrespective of the infection status of the sperm cysts. Group II can be further divided into two subgroups: The first one contains associations with high numbers of heavily Wolbachia-infected sperm cysts while in the second one, Wolbachia is rarely detected in sperm cysts, being mostly present in somatic cells. We conclude that there are three requirements for the expression of CI in a host-Wolbachia association: (a) Wolbachia has to be able to modify sperm (mod(+) genotype), (b) Wolbachia has to infect sperm cysts, and (c) Wolbachia has to be harbored by a permissive host.}, } @article {pmid12803636, year = {2003}, author = {Baudry, E and Bartos, J and Emerson, K and Whitworth, T and Werren, JH}, title = {Wolbachia and genetic variability in the birdnest blowfly Protocalliphora sialia.}, journal = {Molecular ecology}, volume = {12}, number = {7}, pages = {1843-1854}, doi = {10.1046/j.1365-294x.2003.01855.x}, pmid = {12803636}, issn = {0962-1083}, mesh = {Animals ; Base Sequence ; Cluster Analysis ; DNA Primers ; Diptera/*genetics/*microbiology ; *Evolution, Molecular ; *Genetic Variation ; *Geography ; Likelihood Functions ; Molecular Sequence Data ; North America ; Phylogeny ; Polymorphism, Restriction Fragment Length ; Reproduction/genetics ; Sequence Analysis, DNA ; Wolbachia/*physiology ; }, abstract = {Wolbachia are widespread cytoplasmically inherited bacteria that induce various reproductive alterations in host arthropods, including cytoplasmic incompatibility (CI), an incompatibility between sperm and egg that typically results in embryonic death. CI has been invoked as a possible mechanism for reproductive isolation and speciation in arthropods, by restricting gene flow and promoting maintenance (and evolution) of genetic divergence between populations. Here we investigate patterns of Wolbachia infection and nuclear and mitochondrial differentiation in geographical populations of the birdnest blowfly Protocalliphora sialia. Blowflies in western North America are infected with two A-group Wolbachia, with some individuals singly and others doubly infected. Individuals in eastern North America mostly show single infections with a B-group Wolbachia. Populations in the Midwest are polymorphic for infections and show A- or B-group infection. There is a low level of mitochondrial divergence and perfect concordance of mitochondrial haplotype with infection type, suggesting that two Wolbachia-associated selective sweeps of the mitochondrion have occurred in this species. Amplified fragment length polymorphism analysis of nuclear genetic variation shows genetic differentiation between the eastern-Midwestern and western populations. Both Midwestern and eastern flies infected with A-Wolbachia show eastern nuclear genetic profiles. Current results therefore suggest that Wolbachia has not acted as a major barrier to gene flow between western and eastern-Midwestern populations, although some genetic differentiation between A-Wolbachia infected and B-Wolbachia infected individuals in eastern-Midwestern populations cannot be ruled out.}, } @article {pmid12803629, year = {2003}, author = {Dewayne Shoemaker, D and Keller, G and Ross, KG}, title = {Effects of Wolbachia on mtDNA variation in two fire ant species.}, journal = {Molecular ecology}, volume = {12}, number = {7}, pages = {1757-1771}, doi = {10.1046/j.1365-294x.2003.01864.x}, pmid = {12803629}, issn = {0962-1083}, mesh = {Animals ; Ants/*genetics/*microbiology ; Cluster Analysis ; DNA, Mitochondrial/genetics ; *Evolution, Molecular ; *Genetic Variation ; Haplotypes/genetics ; *Phylogeny ; Sequence Analysis, DNA ; Wolbachia/*physiology ; }, abstract = {Wolbachia are endosymbiotic bacteria that infect arthropods. As they are maternally transmitted, the spread of Wolbachia variants within host populations may affect host mtDNA evolution. We sequenced a portion of the mitochondrial cytochrome oxidase I gene from numerous individuals of two Wolbachia-infected fire ant species, Solenopsis invicta and S. richteri, to determine how these bacteria influence patterns of mtDNA variation. As predicted, there was a strong association between Wolbachia strain and host mtDNA lineage within and between these fire ant species. However, there was no consistent association between the presence of Wolbachia and a reduction in mtDNA diversity. Moreover, patterns of mtDNA variation within Wolbachia-infected populations did not differ consistently from neutral expectations, despite our prediction that strong positive selection acting on Wolbachia influences the evolutionary dynamics of other cytoplasmic genomes. Specifically, while values of Tajima's D consistently were less than zero for all six samples of fire ants harbouring Wolbachia, MacDonald-Kreitman tests suggested that the patterns of variation were different from those expected under neutrality in only two of the samples. We conclude that these neutrality tests do not unambiguously reveal a clear effect of Wolbachia infection on patterns of mtDNA variation and substitution in fire ants. Finally, consistent with an earlier study, our data revealed the presence of two divergent mtDNA haplotype lineages and Wolbachia strains within S. invicta. Recognition of these two lineages has important consequences for interpreting patterns of mtDNA evolution and genetic differentiation between conspecific social forms of this species.}, } @article {pmid12801420, year = {2003}, author = {Büttner, DW and Wanji, S and Bazzocchi, C and Bain, O and Fischer, P}, title = {Obligatory symbiotic Wolbachia endobacteria are absent from Loa loa.}, journal = {Filaria journal}, volume = {2}, number = {1}, pages = {10}, pmid = {12801420}, issn = {1475-2883}, abstract = {BACKGROUND: Many filarial nematodes harbour Wolbachia endobacteria. These endobacteria are transmitted vertically from one generation to the next. In several filarial species that have been studied to date they are obligatory symbionts of their hosts. Elimination of the endobacteria by antibiotics interrupts the embryogenesis and hence the production of microfilariae. The medical implication of this being that the use of doxycycline for the treatment of human onchocerciasis and bancroftian filariasis leads to elimination of the Wolbachia and hence sterilisation of the female worms. Wolbachia play a role in the immunopathology of patients and may contribute to side effects seen after antifilarial chemotherapy. In several studies Wolbachia were not observed in Loa loa. Since these results have been doubted, and because of the medical significance, several independent methods were applied to search for Wolbachia in L. loa. METHODS: Loa loa and Onchocerca volvulus were studied by electron microscopy, histology with silver staining, and immunohistology using antibodies against WSP, Wolbachia aspartate aminotransferase, and heat shock protein 60. The results achieved with L. loa and O. volvulus were compared. Searching for Wolbachia, genes were amplified by PCR coding for the bacterial 16S rDNA, the FTSZ cell division protein, and WSP. RESULTS: No Wolbachia endobacteria were discovered by immunohistology in 13 male and 14 female L. loa worms and in numerous L. loa microfilariae. In contrast, endobacteria were found in large numbers in O. volvulus and 14 other filaria species. No intracellular bacteria were seen in electron micrographs of oocytes and young morulae of L. loa in contrast to O. volvulus. In agreement with these results, Wolbachia DNA was not detected by PCR in three male and six female L. loa worms and in two microfilariae samples of L. loa. CONCLUSIONS: Loa loa do not harbour obligatory symbiotic Wolbachia endobacteria in essential numbers to enable their efficient vertical transmission or to play a role in production of microfilariae. Exclusively, the filariae cause the immunopathology of loiasis is patients and the adverse side effects after antifilarial chemotherapy. Doxycycline cannot be used to cure loiais but it probably does not represent a risk for L. loa patients when administered to patients with co-infections of onchocerciasis.}, } @article {pmid12760642, year = {2003}, author = {Gorham, CH and Fang, QQ and Durden, LA}, title = {Wolbachia endosymbionts in fleas (Siphonaptera).}, journal = {The Journal of parasitology}, volume = {89}, number = {2}, pages = {283-289}, doi = {10.1645/0022-3395(2003)089[0283:WEIFS]2.0.CO;2}, pmid = {12760642}, issn = {0022-3395}, mesh = {Animals ; Cats ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Dogs ; Female ; Georgia ; Male ; New York ; Opossums ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Sciuridae ; Sequence Alignment ; Sigmodontinae ; Siphonaptera/classification/*microbiology ; *Symbiosis ; Wolbachia/classification/genetics/isolation & purification/*physiology ; }, abstract = {Intracellular endosymbionts, Wolbachia spp., have been reported in many different orders of insects and in nematodes but not previously in fleas. This is the first conclusive report of Wolbachia spp. within members of the Siphonaptera. Using nested polymerase chain reaction (PCR) targeting of the 16S ribosomal RNA gene, we screened for Wolbachia spp. in fleas collected from 3 counties in Georgia and 1 in New York. The prevalence of Wolbachia spp. detected varied among the 6 different species screened: 21% in the cat flea Ctenocephalides felis (n = 604), 7% in the dog flea C. canis (n = 28), 25% in Polygenus gwyni (n = 8), 80% in Orchopeas howardi (n = 15), 94% in Pulex simulans (n = 255), and 24% in the sticktight flea Echidnophaga gallinacea (n = 101). Wolbachia spp. infection in fleas was confirmed by sequencing positive PCR products, comparing sequenced 16S ribosomal DNA (rDNA) with Wolbachia spp. sequences in GenBank using BLAST search, and subjecting sequence data to phylogenetic analysis. For further confirmation, 16S rDNA-positive samples were reamplified using the wsp gene.}, } @article {pmid12753632, year = {2003}, author = {Volkmann, L and Fischer, K and Taylor, M and Hoerauf, A}, title = {Antibiotic therapy in murine filariasis (Litomosoides sigmodontis): comparative effects of doxycycline and rifampicin on Wolbachia and filarial viability.}, journal = {Tropical medicine & international health : TM & IH}, volume = {8}, number = {5}, pages = {392-401}, doi = {10.1046/j.1365-3156.2003.01040.x}, pmid = {12753632}, issn = {1360-2276}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/therapeutic use ; Drug Therapy, Combination/therapeutic use ; Female ; Filariasis/complications/*drug therapy/pathology ; Filarioidea/drug effects/growth & development ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophil Infiltration/drug effects ; Polymerase Chain Reaction ; Rickettsiaceae Infections/complications/*drug therapy/pathology ; Rifampin/therapeutic use ; Symbiosis/drug effects ; Wolbachia/*drug effects/growth & development ; }, abstract = {The symbiosis of filarial nematodes and rickettsial Wolbachia endobacteria has been exploited as a target for antibiotic therapy of filariasis. Depletion of Wolbachia after tetracycline treatment results in filarial sterility because of interruption of embryogenesis and inhibits larval development and adult worm viability. The aim of this study was to investigate if antibiotic intervention of BALB/c mice infected with the rodent filaria Litomosoides sigmodontis with rifampicin or the combination of rifampicin and doxycycline can be used to shorten the treatment period. Both regimens, when given over a period of 14 days initiated with infection, were sufficient to deplete Wolbachia as evidenced by immunohistology and semiquantitative PCR. Worm development and filarial load were significantly reduced in experiments followed up until 63 days p.i. The therapy inhibited embryogenesis and led to filarial sterility. In contrast, treatment with doxycycline alone for 21 days led only to a modest reduction of Wolbachia, filarial growth retardation, worm viability and fertility. In conclusion, the combination of antirickettsial drugs could be used as a suitable tool to explore the minimum duration of therapy required for the depletion of Wolbachia in parasitized hosts subsequent to the onset of patency in human and animal filariasis and the prevention of adverse reactions in human infections.}, } @article {pmid12753223, year = {2003}, author = {Kittayapong, P and Jamnongluk, W and Thipaksorn, A and Milne, JR and Sindhusake, C}, title = {Wolbachia infection complexity among insects in the tropical rice-field community.}, journal = {Molecular ecology}, volume = {12}, number = {4}, pages = {1049-1060}, doi = {10.1046/j.1365-294x.2003.01793.x}, pmid = {12753223}, issn = {0962-1083}, mesh = {Animals ; Base Sequence ; Cluster Analysis ; Environment ; Insecta/*microbiology ; Molecular Sequence Data ; Oryza/physiology ; *Phylogeny ; Thailand ; Wolbachia/classification/*genetics/*physiology ; }, abstract = {Wolbachia are a group of intracellular bacteria that cause reproductive alterations in their arthropod hosts. Widely discordant host and Wolbachia phylogenies indicate that horizontal transmission of these bacteria among species sometimes occurs. A likely means of horizontal transfer is through the feeding relations of organisms within communities. Feeding interactions among insects within the rice-field insect community have been well documented in the past. Here, we present the results of a polymerase chain reaction-based survey and phylogenetic analysis of Wolbachia strains in the rice-field insect community of Thailand. Our field survey indicated that 49 of 209 (23.4%) rice-field insect species were infected with Wolbachia. Of the 49 infected species, 27 were members of two feeding complexes: (i) a group of 13 hoppers preyed on by 2 mirid species and parasitized by a fly species, and (ii) 2 lepidopteran pests parasitized by 9 wasp species. Wolbachia strains found in three hoppers, Recilia dorsalis, Nephotettix malayanus and Nisia nervosa, the two mirid predators, Cyrtorhinus lividipennis and Tytthus chinensis, and the fly parasitoid, Tomosvaryella subvirescens, were all in the same Wolbachia clade. In the second complex, the two lepidopteran pests, Cnaphalocrocis medinalis and Scirpophaga incertulas, were both infected with Wolbachia from the same clade, as was the parasitoid Tropobracon schoenobii. However, none of the other infected parasitoid species in this feeding complex was infected by Wolbachia from this clade. Mean (+/- SD) genetic distance of Wolbachia wsp sequences among interacting species pairs of the hopper feeding complex (0.118 +/- 0.091 nucleotide sequence differences), but not for the other two complexes, was significantly smaller than that between noninteracting species pairs (0.162 +/- 0.079 nucleotide sequence differences). Our results suggest that some feeding complexes, such as the hopper complex described here, could be an important means by which Wolbachia spreads among species within arthropod communities.}, } @article {pmid12750334, year = {2003}, author = {Bordenstein, SR and Uy, JJ and Werren, JH}, title = {Host genotype determines cytoplasmic incompatibility type in the haplodiploid genus Nasonia.}, journal = {Genetics}, volume = {164}, number = {1}, pages = {223-233}, pmid = {12750334}, issn = {0016-6731}, mesh = {Animals ; Crosses, Genetic ; Female ; Genotype ; Host-Parasite Interactions/*genetics ; Male ; Ovum/*metabolism ; Spermatozoa/*metabolism ; Wasps/*genetics/microbiology ; Wolbachia/*genetics/metabolism ; }, abstract = {In haplodiploid species, Wolbachia-induced cytoplasmic incompatibility (CI) can be expressed in one of two ways: as a "conversion" of diploid fertilized eggs into haploid males or as embryonic mortality. Here we describe CI-type variation within the parasitic wasp genus Nasonia and genetically analyze the basis of this variation. We reach four main conclusions: (i) CI is expressed primarily as conversion in N. vitripennis, but as embryonic mortality in the sibling species N. giraulti and N. longicornis; (ii) the difference in CI type between N. giraulti (mortality) and N. vitripennis (conversion) is determined by host nuclear genotype rather than by Wolbachia differences; (iii) N. vitripennis "conversion genes" are recessive in hybrid females; and (iv) a difference in CI level between the sibling species N. giraulti and N. longicornis is due to the different Wolbachia infections in the species rather than to the host genotype. These results show that host nuclear genes can influence the type of CI present in a species. On the basis of these findings, we propose a model for how different CI types evolve in haplodiploids due to selection on nuclear genes modifying CI.}, } @article {pmid12750316, year = {2003}, author = {Jiggins, FM}, title = {Male-killing Wolbachia and mitochondrial DNA: selective sweeps, hybrid introgression and parasite population dynamics.}, journal = {Genetics}, volume = {164}, number = {1}, pages = {5-12}, pmid = {12750316}, issn = {0016-6731}, mesh = {Animals ; Butterflies/*genetics/microbiology ; *DNA, Mitochondrial ; Female ; Genetic Variation ; *Hybridization, Genetic ; Likelihood Functions ; Male ; Molecular Sequence Data ; Phylogeny ; Population Dynamics ; Selection, Genetic ; Wolbachia/*genetics ; }, abstract = {Mitochondrial DNA (mtDNA) sequences are widely used as neutral genetic markers in insects. However, patterns of mtDNA variability are confounded by the spread of maternally transmitted parasites, which are genetically linked to the mitochondria. We have investigated these effects in the butterflies Acraea encedon (which is host to two strains of male-killing Wolbachia bacteria) and A. encedana (which is host to one strain). Within a population, the mitochondria are in linkage disequilibrium with the different male-killers. Furthermore, there has been a recent selective sweep of the mtDNA, which has led to the loss of mitochondrial variation within populations and erased any geographical structure. We also found that one of the male-killers, together with the associated mtDNA, has introgressed from A. encedana into A. encedon within the last 16,000 years. Interestingly, because butterflies are female heterogametic, this will presumably have also led to the introgression of genes on the W sex chromosome. Finally, in A. encedon the mitochondria in uninfected females are unaltered by the spread of the male-killer and have diverse, geographically structured mtDNA. This means we can reject the hypothesis that the male-killer is at a stable equilibrium maintained by imperfect transmission of the bacterium. Instead, some other form of balancing selection may be maintaining uninfected females in the population and preventing the species from going extinct due to a shortage of males.}, } @article {pmid12748849, year = {2003}, author = {Grobusch, MP and Kombila, M and Autenrieth, I and Mehlhorn, H and Kremsner, PG}, title = {No evidence of Wolbachia endosymbiosis with Loa loa and Mansonella perstans.}, journal = {Parasitology research}, volume = {90}, number = {5}, pages = {405-408}, pmid = {12748849}, issn = {0932-0113}, mesh = {Animals ; Female ; Humans ; Loa/isolation & purification/*microbiology/ultrastructure ; Loiasis/parasitology ; Male ; Mansonella/isolation & purification/*microbiology/ultrastructure ; Mansonelliasis/parasitology ; Microfilariae ; Microscopy, Electron ; Polymerase Chain Reaction ; Symbiosis ; Wolbachia/isolation & purification/*physiology ; }, abstract = {Endosymbiotic Wolbachia bacteria from different filarial species, including major pathogens of humans such as Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus, seem to play an important role in the development, viability and fertility of these worms. Wolbachia trigger inflammatory host responses as well as adverse reactions against standard treatment regimens and are therefore under investigation as novel treatment targets. We investigated whether Wolbachia are also endosymbiotic in Loa loa and Mansonella perstans. In both male and female adult L. loa, we found no evidence of bacteria by light or transmission electron microscopy. Furthermore, Wolbachia-specific PCR was negative in both L. loa and M. perstans microfilariae. The absence of Wolbachia in both filarial species therefore discourages the use of antibiotics as an adjunct or alternative approach to current treatment concepts for both loiasis and mansonelliasis perstans.}, } @article {pmid12743802, year = {2003}, author = {Fischer, P and Bonow, I and Büttner, DW and Kamal, IH and Liebau, E}, title = {An aspartate aminotransferase of Wolbachia endobacteria from Onchocerca volvulus is recognized by IgG1 antibodies from residents of endemic areas.}, journal = {Parasitology research}, volume = {90}, number = {1}, pages = {38-47}, pmid = {12743802}, issn = {0932-0113}, mesh = {Amino Acid Sequence ; Animals ; Antibodies, Bacterial/*immunology ; Aspartate Aminotransferases/blood/genetics/*immunology/isolation & purification ; DNA, Bacterial/genetics/isolation & purification ; *Endemic Diseases ; Female ; Humans ; Immunoglobulin G/*immunology ; Immunohistochemistry ; Molecular Sequence Data ; Onchocerca volvulus/*microbiology ; Onchocerciasis/*immunology ; Rabbits ; Sequence Alignment ; Seroepidemiologic Studies ; Staining and Labeling ; Wolbachia/enzymology/*immunology ; }, abstract = {Wolbachia are intracellular alpha-proteobacteria, closely related to Rickettsia, that infect various arthropods and filarial parasites. In the present study, the cDNA encoding the aspartate aminotransferase (AspAT) of Wolbachia from the human pathogenic filarial parasite Onchocerca volvulus (Ov-WolAspAT) was identified. At the amino acid level, the identity of the Ov-WolAspAT was 56% to Rickettsia prowazekii AspAT and 54% to the AspAT of the nitrogen-fixing bacterium Sinorhizobium meliloti, but the highest degree of identity was found to the putative AspAT of Wolbachia from Brugia malayi and Drosophila melanogaster (85%). All of these bacterial AspATs are members of the AspAT subclass Ib. A 35 kDa fragment of the Ov-WolAspAT was expressed in Escherichia coli, and immunolocalization using polyclonal antibodies against this antigen revealed that Ov-WolAspAT is present in a considerable proportion of the Wolbachia from O. volvulus, as well as in the endobacteria of several other filarial parasites. Western blot analysis using recombinant Ov-WolAspAT as antigen showed that IgG1 antibodies were present in 70 (51%) individuals living in areas endemic for O. volvulus, B. malayi or Wuchereria bancrofti and no IgG4 or IgE antibodies were found. Among 40 sera of persons from Uganda and Liberia who were putatively not infected with human filarial parasites, 11 (28%) individuals presented IgG1 antibodies, while none of the 33 sera from healthy Europeans and none of the 14 sera from patients with proven Rickettsia or Brucella infections reacted with the antigen. These results also show that an intracellular protein of Wolbachia endobacteria (WolAspAT) acts as antigen in human filariasis.}, } @article {pmid12717985, year = {2003}, author = {Watanabe, H and Mitobe, J}, title = {[Possible bacterial etiologies for diseases of unknown origin].}, journal = {Nihon rinsho. Japanese journal of clinical medicine}, volume = {61 Suppl 3}, number = {}, pages = {285-289}, pmid = {12717985}, issn = {0047-1852}, mesh = {Actinobacteria ; Angiomatosis, Bacillary/*etiology ; Animals ; Bartonella Infections/complications ; Bartonella henselae ; *Campylobacter jejuni ; Gastritis, Atrophic/*etiology ; Gram-Positive Bacterial Infections/*complications ; Guillain-Barre Syndrome/*etiology ; Helicobacter Infections/*complications ; *Helicobacter pylori ; Humans ; Onchocerca volvulus/microbiology ; Onchocerciasis/etiology ; Propionibacterium acnes ; Sarcoidosis/*etiology ; Stomach Neoplasms/*etiology ; Whipple Disease/*etiology ; Wolbachia ; }, } @article {pmid12710612, year = {2003}, author = {Taillardat-Bisch, AV and Raoult, D and Drancourt, M}, title = {RNA polymerase beta-subunit-based phylogeny of Ehrlichia spp., Anaplasma spp., Neorickettsia spp. and Wolbachia pipientis.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {53}, number = {Pt 2}, pages = {455-458}, doi = {10.1099/ijs.0.02411-0}, pmid = {12710612}, issn = {1466-5026}, mesh = {Anaplasma/*classification/enzymology/genetics ; Bacterial Proteins/genetics ; DNA-Directed RNA Polymerases/*genetics ; Ehrlichia/*classification/enzymology/genetics ; Molecular Sequence Data ; Neorickettsia/*classification/enzymology/genetics ; Phylogeny ; RNA, Ribosomal, 16S/analysis/genetics ; Sequence Analysis, DNA ; Wolbachia/*classification/enzymology/genetics ; }, abstract = {Sequence analysis of rpoB, the gene encoding the beta-subunit of RNA polymerase, was used in a phylogenetic investigation of nine species from the genera Ehrlichia, Neorickettsia, Wolbachia and Anaplasma. The complete nucleotide sequences obtained for Anaplasma phagocytophilum (HGE agent), Ehrlichia chaffeensis, Neorickettsia sennetsu, Neorickettsia risticii, Anaplasma marginale and Wolbachia pipientis were amongst the longest rpoB sequences in GenBank and ranged from 4074 bp for N. sennetsu to 4311 bp for W. pipientis. Additional partial rpoB sequences were obtained for Ehrlichia canis, Ehrlichia ruminantium and Ehrlichia muris. Identical phylogenetic trees were inferred from multiple sequence alignments of the nucleotide sequences and the derived amino acid sequences using either distance, maximum-likelihood or parsimony methods. This study confirms the phylogeny previously inferred from sequence analyses of the 16S rRNA gene, groESL and gltA and allows the confirmation of four monophyletic clades. The rpoB nucleotide sequences were more variable than the 16S rRNA gene and groESL sequences at the species level.}, } @article {pmid12709338, year = {2003}, author = {Fenollar, F and Maurin, M and Raoult, D}, title = {Wolbachia pipientis growth kinetics and susceptibilities to 13 antibiotics determined by immunofluorescence staining and real-time PCR.}, journal = {Antimicrobial agents and chemotherapy}, volume = {47}, number = {5}, pages = {1665-1671}, pmid = {12709338}, issn = {0066-4804}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Fluorescent Antibody Technique ; Microbial Sensitivity Tests ; *Polymerase Chain Reaction ; Rabbits ; Wolbachia/*drug effects/genetics/growth & development ; }, abstract = {Wolbachia spp. are strict intracellular bacteria that infect a wide range of arthropods and filarial nematodes. Filarial nematodes are important causes of human diseases. There is increasing evidence that Wolbachia spp. influence important functions in the biology of the hosts, specifically, infertility. Preliminary experiments with humans and animals have suggested that antibiotics with activity against Wolbachia may help to treat filariasis. In this study, we determined using a real-time quantitative PCR assay the growth kinetics of a strain of Wolbachia pipientis from a mosquito grown in Aa23 cells. The doubling time was estimated to be 14 h. We then determined the susceptibilities of this strain to 13 antibiotics by two methods: an immunofluorescent-antibody test and a real-time quantitative PCR assay. Both techniques gave similar results. Doxycycline and rifampin were the most effective compounds, with MICs of 0.125 and 0.06 to 0.125 micro g/ml, respectively. Fluoroquinolones were less effective, with MICs of 2 to 4 micro g/ml for ciprofloxacin, 2 micro g/ml for ofloxacin, and 1 micro g/ml for levofloxacin. beta-Lactams (penicillin G, amoxicillin, ceftriaxone) were not effective at concentrations up to 128 micro g/ml. The MIC of erythromycin was >32 micro g/ml, whereas that of telithromycin was 8 micro g/ml. Other antibiotic compounds were bacteriostatic only at high concentrations, including gentamicin, co-trimoxazole, and thiamphenicol. The real-time PCR assay was a convenient and reliable technique for determination of the antibiotic susceptibilities of WOLBACHIA: It may help in the future to simplify antibiotic susceptibility testing of strict intracellular pathogens.}, } @article {pmid12706439, year = {2003}, author = {Hoerauf, A and Mand, S and Volkmann, L and Büttner, M and Marfo-Debrekyei, Y and Taylor, M and Adjei, O and Büttner, DW}, title = {Doxycycline in the treatment of human onchocerciasis: Kinetics of Wolbachia endobacteria reduction and of inhibition of embryogenesis in female Onchocerca worms.}, journal = {Microbes and infection}, volume = {5}, number = {4}, pages = {261-273}, doi = {10.1016/s1286-4579(03)00026-1}, pmid = {12706439}, issn = {1286-4579}, mesh = {Animals ; Anti-Bacterial Agents/pharmacokinetics/*therapeutic use ; Doxycycline/pharmacokinetics/*therapeutic use ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry/methods ; Ivermectin/pharmacology ; Male ; Microfilariae/drug effects/growth & development ; Onchocerca volvulus/*drug effects/embryology/growth & development/pathogenicity ; Onchocerciasis/*drug therapy/epidemiology/pathology ; Polymerase Chain Reaction/methods ; Spermatogenesis/drug effects ; Spermatozoa/drug effects ; Treatment Outcome ; Wolbachia/*drug effects/metabolism ; }, abstract = {Recently, experts have warned that mass treatment with ivermectin alone may not interrupt the transmission of Onchocerca. Hence, additional drugs are needed, such as antibiotics acting on symbiotic endobacteria of the filariae, the causative agents of onchocerciasis. Based on animal experiments, human onchocerciasis was treated with doxycycline, and preliminary observations published in 2001 in The Lancet showed sterility in female worms by depletion and marked reduction in symbiotic Wolbachia endobacteria from the filariae. Here, a detailed kinetic analysis of the features of the worms, following administration or not of doxycycline to the patients is reported. Sixty-three onchocerciasis patients in Ghana were treated with 100 mg doxycycline daily for 6 weeks and 2 or 6 months later with ivermectin. Onchocercomas were extirpated 2, 6, 11 and 18 months after the onset of treatment and the filariae were examined by immunohistology and PCR. The analysis showed: (i) progressive depletion of Wolbachia from adult worms and microfilariae by doxycycline over a period of 6 months; (ii) inhibition of embryogenesis by doxycycline after 6 months with respect to all embryo stages followed by decline in microfilariae after 11 months; (iii) reduction in spermatozoa in the female genital tract by doxycycline, whereas spermiogenesis was only partly reduced after 11 and 18 months; (iv) no relevant macro- or microfilaricidal activity; (v) depletion/marked reduction in endobacteria and inhibition of embryogenesis were sustained until 18 months after doxycycline and 12 months after co-administration of ivermectin; (vi) no severe adverse side effects were seen. Due to its long-lasting inhibition of embryogenesis, doxycycline presents an additional strategy for the treatment of onchocerciasis and control of Onchocerca microfilariae transmission. Extension of the existing registration will not require much time or high cost. Treatment of individual patients can be considered immediately.}, } @article {pmid12701381, year = {2002}, author = {Baldo, L and Bartos, JD and Werren, JH and Bazzocchi, C and Casiraghi, M and Panelli, S}, title = {Different rates of nucleotide substitutions in Wolbachia endosymbionts of arthropods and nematodes: arms race or host shifts?.}, journal = {Parassitologia}, volume = {44}, number = {3-4}, pages = {179-187}, pmid = {12701381}, issn = {0048-2951}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Base Sequence ; Codon/genetics ; DNA-Binding Proteins/genetics ; *Evolution, Molecular ; Fushi Tarazu Transcription Factors ; Genes, Bacterial ; Homeodomain Proteins/genetics ; Likelihood Functions ; *Mutagenesis ; Nematoda/*microbiology ; Phylogeny ; Selection, Genetic ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid ; Species Specificity ; Wolbachia/*genetics/ultrastructure ; }, abstract = {The genus Wolbachia encompasses intracellular bacteria found in arthropods and in filarial nematodes. In arthropods, Wolbachia is primarily a reproductive parasite and shows relatively frequent horizontal transfer between host species, while in nematodes it appears to be a mutualist and is strictly vertically transmitted. We can expect that different selective pressures are acting on their genomes. Here we present an analysis of three Wolbachia genes, wsp, ftsZ and dnaA. In wsp of arthropod Wolbachia, an excess of non-synonymous substitutions was observed, providing evidence for positive selection. In nematode Wolbachia, no evidence for positive selection was found. Pressure for amino acid variation in wsp of arthropod Wolbachia could derive either from an arms race with the host or from the occurrence of more frequent hosts shifts due to horizontal transmission. In nematode Wolbachia, the lack of positively selected sites could result from the absence of an arms race, or from the homogeneity of the biochemical environment they exist in (ensured by strict vertical transmission). In ftsZ minor differences in substitution patterns were observed between arthropod and nematode Wolbachia, only in the 3'-portion of the gene. dnaA showed comparable patterns of variation in both lineages, with evidence for strong conservation.}, } @article {pmid12693853, year = {2003}, author = {Hirunkanokpun, S and Kittayapong, P and Cornet, JP and Gonzalez, JP}, title = {Molecular evidence for novel tick-associated spotted fever group rickettsiae from Thailand.}, journal = {Journal of medical entomology}, volume = {40}, number = {2}, pages = {230-237}, doi = {10.1603/0022-2585-40.2.230}, pmid = {12693853}, issn = {0022-2585}, mesh = {Animals ; Base Sequence ; Cattle/microbiology ; DNA Primers ; Dogs/microbiology ; Genotype ; Humans ; Phylogeny ; Rats/microbiology ; Rickettsia/*classification/*genetics/isolation & purification ; Rickettsia Infections/*epidemiology ; Thailand/epidemiology ; Ticks/*microbiology ; }, abstract = {Ticks are of considerable medical and veterinary importance because they directly harm the host through their feeding action and indirectly through vectoring many bacterial pathogens. Despite many ticks being known from Thailand, very little is known about the bacteria they may harbor. We report here the results of a survey of tick-associated bacteria in Thailand. A total of 334 individuals representing 14 species of ticks in five genera were collected from 10 locations in Thailand and were examined for the human pathogens, Borrelia, Francisella, Rickettsia, and the common arthropod endosymbionts, Wolbachia, by polymerase chain reaction (PCR) assay using specific primers. Rickettsial DNA was detected in 30% (9/30) of Amblyomma testudinarium (Koch, 1844) collected from Khao Yai National Park, Nakhon Nayok Province and 16.84% (16/95) of Hemaphysalis ornithophila (Hoogstraal and Kohls, 1959) collected from Khao Yai National Park, Nakhon Nayok Province and Khao Ang Rue Nai Wildlife Sanctuary, Chachoengsao Province. Rickettsial DNA was not detected in any of the other tick species and no DNA of Borrelia, Francisella, or Wolbachia was detected in any of 14 tick species. Phylogenetic relationships among the rickettsiae detected in this study and those of other rickettsiae were inferred from comparison of sequences of the 17-kDa antigen gene, the citrate synthase gene (gltA), and the 190-kDa outer membrane protein gene (ompA). Results indicated that the three Thai rickettsiae detected in this study represent new rickettsial genotypes and form a separate cluster among the spotted fever group rickettsiae.}, } @article {pmid12693838, year = {2003}, author = {Rasgon, JL and Styer, LM and Scott, TW}, title = {Wolbachia-induced mortality as a mechanism to modulate pathogen transmission by vector arthropods.}, journal = {Journal of medical entomology}, volume = {40}, number = {2}, pages = {125-132}, doi = {10.1603/0022-2585-40.2.125}, pmid = {12693838}, issn = {0022-2585}, support = {GM-20092/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropod Vectors/microbiology/*physiology ; Culex/microbiology/physiology ; Female ; Male ; Reproduction ; Wolbachia/*pathogenicity ; }, abstract = {Insecticide resistance and absence of clinical cures or vaccines for many vector-borne diseases has stimulated interest in using genetically modified arthropod vectors for disease control. Current transgenic strategies focus on vector susceptibility to pathogen infection, which is an inefficient target for pathogen transmission interference. Manipulation of vector survival is theoretically more effective, resulting in larger reductions in the expected number of human infections. A hypothetical method to manipulate vector survival is to drive mortality-inducing Wolbachia into populations. For varying patterns and degrees of induced mortality, we outline the conditions under which virulent Wolbachia introductions into vector populations are expected to succeed and quantify the resultant reduction in pathogen transmission. The most critical component to the success of this strategy is the pattern of induced mortality. For operationally feasible introductions, induced mortality must be delayed until after vector reproduction begins. If this condition is not met, introduction thresholds become exceedingly high, ranging from approximately 40% to 90% of the total adult population. Delayed induced mortality patterns can reduce introduction thresholds to approximately 15-45% of the total adult population. Reduction in cytoplasmic incompatibility with male age has negligible effects on introduction success regardless of the induced mortality pattern. Under proper circumstances, symbiont-induced manipulation of vector survival can theoretically result in up to 100% reduction in pathogen transmission, depending on Wolbachia parameters, magnitude and pattern of induced mortality, and duration of pathogen incubation in the vector. Our results indicate that a broadening of the current paradigm for genetic manipulation of vectors to parameters other than arthropod vector competence is justified and will reveal new research possibilities for vector-borne disease control.}, } @article {pmid12684759, year = {2003}, author = {Hoerauf, A and Mand, S and Fischer, K and Kruppa, T and Marfo-Debrekyei, Y and Debrah, AY and Pfarr, KM and Adjei, O and Büttner, DW}, title = {Doxycycline as a novel strategy against bancroftian filariasis-depletion of Wolbachia endosymbionts from Wuchereria bancrofti and stop of microfilaria production.}, journal = {Medical microbiology and immunology}, volume = {192}, number = {4}, pages = {211-216}, pmid = {12684759}, issn = {0300-8584}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/administration & dosage/pharmacology/*therapeutic use ; Doxycycline/administration & dosage/pharmacology/*therapeutic use ; Elephantiasis, Filarial/*drug therapy/*parasitology ; Female ; Filaricides/administration & dosage/pharmacology/therapeutic use ; Ghana ; Humans ; Ivermectin/administration & dosage/pharmacology/therapeutic use ; Male ; Microfilariae/drug effects ; Middle Aged ; Parasitemia/drug therapy ; Symbiosis ; Wolbachia/*drug effects/growth & development ; Wuchereria bancrofti/drug effects/growth & development/*microbiology ; }, abstract = {Chemotherapy of onchocerciasis by doxycycline, which targets symbiotic Wolbachia endobacteria, has been shown to result in a long-term sterility of adult female worms and corresponding absence of microfilariae. It represents an additional chemotherapeutic approach. The aim of this study was to determine whether a similar regimen would also show efficacy against Wuchereria bancrofti. Ghanaian individuals (n=93) with lymphatic filariasis and a minimum microfilaremia of 40 microfilariae/ml were included in a treatment study consisting of four arms: (1) doxycycline 200 mg/day for 6 weeks; (2) doxycycline as in (1), followed by a single dose of ivermectin after 4 months; (3) ivermectin only; or (4) no treatment during observation period of 1 year (ivermectin at the end of the study). Doxycycline treatment resulted in a 96% loss of Wolbachia, as determined by real time PCR from microfilariae. After 12 months, doxycycline had led to a 99% reduction of microfilaremia when given alone, and to a complete amicrofilaremia together with ivermectin. In contrast, after ivermectin treatment alone a significant presence of microfilariae remained (9% compared to pretreatment), as known from other studies. This study shows that doxycycline is also effective in depleting Wolbachia from W. bancrofti. It is likely that the mechanism of doxycycline is similar to that in other filarial species, i.e., a predominant blockade of embryogenesis, leading to a decline of microfilariae according to their half-life. This could render doxycycline treatment an additional tool for the treatment of microfilaria-associated diseases in bancroftian filariasis, such as tropical pulmonary eosinophilia and microfiluria.}, } @article {pmid12683975, year = {2003}, author = {Charlat, S and Hurst, GD and Merçot, H}, title = {Evolutionary consequences of Wolbachia infections.}, journal = {Trends in genetics : TIG}, volume = {19}, number = {4}, pages = {217-223}, doi = {10.1016/S0168-9525(03)00024-6}, pmid = {12683975}, issn = {0168-9525}, mesh = {Animals ; Arthropods/genetics/*microbiology ; *Biological Evolution ; Cell Cycle/genetics/physiology ; Cell Differentiation/genetics/physiology ; Germ Cells ; Nematoda/genetics/*microbiology ; Sex Determination Processes ; Wolbachia/*genetics/pathogenicity ; }, abstract = {The past decade has revealed the bacterium Wolbachia as the most widespread symbiont of arthropods and nematodes. Behind this evolutionary success is an remarkable variety of effects on host biology, ranging from manipulation of reproduction in favor of females to more classical mutualistic interactions. Here we discuss the potential of Wolbachia for promoting evolutionary changes in its hosts.}, } @article {pmid12683537, year = {2003}, author = {Vavre, F and Fouillet, P and Fleury, F}, title = {Between- and within-host species selection on cytoplasmic incompatibility-inducing Wolbachia in haplodiploids.}, journal = {Evolution; international journal of organic evolution}, volume = {57}, number = {2}, pages = {421-427}, doi = {10.1111/j.0014-3820.2003.tb00275.x}, pmid = {12683537}, issn = {0014-3820}, mesh = {Alleles ; Animals ; Biological Evolution ; Cytoplasm/microbiology ; Diploidy ; Female ; Genes, Insect ; Genetic Variation ; Haploidy ; Insecta/*genetics/*microbiology ; Male ; Models, Biological ; Reproduction/genetics ; Species Specificity ; Symbiosis/genetics/physiology ; Wolbachia/genetics/pathogenicity/*physiology ; }, abstract = {The most common effect of the endosymbiont Wolbachia is cytoplasmic incompatibility (CI), a form of postzygotic reproductive isolation that occurs in crosses where the male is infected by at least one Wolbachia strain that the female lacks. We revisited two puzzling features of Wolbachia biology: how Wolbachia can invade a new species and spread among populations, and how the association, once established in a host species, can evolve, with emphasis on the possible process of infection loss. These questions are particularly relevant in haplodiploid species, where males develop from unfertilized eggs, and females from fertilized eggs. When CI occurs in such species, fertilized eggs either die (female mortality type: FM), or develop into males (male development type: MD), raising one more question: how transition among CI types is possible. We reached the following conclusions: (1) the FM type is a better invader and should be retained preferentially after a new host is captured; (2) given the assumptions of the models, FM and MD types are selected on neither the bacterial side nor the host side; (3) selective pressures acting on both partners are more or less congruent in the FM type, but divergent in the MD type; (4) host and symbiont evolution can drive infection to extinction for all CI types, but the MD type is more susceptible to the phenomenon; and (5) under realistic conditions, transition from MD to FM type is possible. Finally, all these results suggest that the FM type should be more frequent than the MD type, which is consistent with the results obtained so far in haplodiploids.}, } @article {pmid12679529, year = {2003}, author = {Reuter, M and Keller, L}, title = {High levels of multiple Wolbachia infection and recombination in the ant Formica exsecta.}, journal = {Molecular biology and evolution}, volume = {20}, number = {5}, pages = {748-753}, doi = {10.1093/molbev/msg082}, pmid = {12679529}, issn = {0737-4038}, mesh = {Animals ; Ants/*genetics/*microbiology/physiology ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; *Biological Evolution ; *Genetic Variation ; Molecular Sequence Data ; Polymorphism, Restriction Fragment Length ; Recombination, Genetic/genetics ; Reproduction/physiology ; Sequence Analysis, DNA ; Switzerland ; Symbiosis ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia bacteria are intracellular symbionts of many arthropod species. Their spread through host populations is promoted by drastic alterations imposed on their hosts' reproductive physiology. In the present study, we analyzed the association between Wolbachia strains and host mitochondrial haplotypes in a Swiss population of the ant Formica exsecta. In this species, female dispersal is extremely limited and the mitochondrial haplotypes are strongly differentiated between and within subpopulations. Our study revealed exceptionally high levels of multiple infection, with all ants harboring four or five distinct Wolbachia strains. Four of these strains were present in all ants analyzed. A fifth strain was associated with only three of the five mitochondrial haplotypes. An analysis of the Wolbachia gene wsp further revealed an unexpected high rate of recombination, with three of the five Wolbachia strains appearing to have arisen by homologous recombination.}, } @article {pmid12675371, year = {2003}, author = {Vala, F and Van Opijnen, T and Breeuwer, JA and Sabelis, MW}, title = {Genetic conflicts over sex ratio: mite-endosymbiont interactions.}, journal = {The American naturalist}, volume = {161}, number = {2}, pages = {254-266}, doi = {10.1086/345856}, pmid = {12675371}, issn = {0003-0147}, mesh = {Animals ; Biological Evolution ; Female ; Host-Parasite Interactions ; Litter Size ; Male ; *Reproduction ; *Sex Ratio ; *Symbiosis ; Tetranychidae/genetics/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Nucleocytoplasmic genetic conflicts arise as a result of asymmetric transmission of cytoplasmic and nuclear genes. Spread of a cytoplasmic element promoting female-biased sex ratios creates selection on nuclear genes for mechanisms that decrease the bias. Here we investigate the conflict over sex ratio between the cytoplasmic bacterium Wolbachia and the two-spotted spider mite Tetranychus urticae Koch. We show that, first, infected females produce significantly more female-biased sex ratios than uninfected (cured) females. Second, this effect is not due to parthenogenesis, male killing, or feminization, phenotypes commonly associated with infection by Wolbachia. Third, sex ratio is a trait with a heritable component in this species; thus, it can evolve under selection. Fourth, the sex ratio produced by uninfected (cured) females changes over time, approaching the sex ratio produced by females from the infected culture. On the basis of these results, we suggest that after sex ratio manipulation by Wolbachia, a host compensatory mechanism evolved that allows infected females to produce the sex ratio favored by nuclear genes. We discuss the evolution of "mutualism" with respect to the evolution of host mechanisms that compensate for effects induced by vertically transmitted "parasites."}, } @article {pmid12669810, year = {2003}, author = {Belshaw, R and Quicke, DL}, title = {The cytogenetics of thelytoky in a predominantly asexual parasitoid wasp with covert sex.}, journal = {Genome}, volume = {46}, number = {1}, pages = {170-173}, doi = {10.1139/g02-112}, pmid = {12669810}, issn = {0831-2796}, mesh = {Animals ; *Cytogenetic Analysis ; Female ; Ovum/cytology ; Reproduction, Asexual/*genetics ; Wasps/*genetics/microbiology ; Wolbachia/physiology ; }, abstract = {Asexual lineages in the parasitoid wasp Lysiphlebus fabarum (Hymenoptera: Braconidae: Aphidiinae) have previously been shown to have occasionally undergone sexual reproduction and recombination with males from related sexual populations. In the present study, the cytogenetic system of asexual females in this species is shown by 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) staining to be central fusion automixis. This system has the potential to allow occasional sex and recombination without leading to an elevation of ploidy and with the maintenance of at least some heterozygosity. No evidence of the bacterial symbiont Wolbachia was found, and the observed system was compared with that in parasitoid wasps where asexuality is the result of Wolbachia infection.}, } @article {pmid12655178, year = {2003}, author = {Ikeda, T and Ishikawa, H and Sasaki, T}, title = {Regulation of Wolbachia density in the Mediterranean flour moth, Ephestia kuehniella, and the almond moth, Cadra cautella.}, journal = {Zoological science}, volume = {20}, number = {2}, pages = {153-157}, doi = {10.2108/zsj.20.153}, pmid = {12655178}, issn = {0289-0003}, mesh = {Animals ; Colony Count, Microbial ; Embryo, Nonmammalian/microbiology ; Female ; Host-Parasite Interactions ; Larva/microbiology ; Lepidoptera/embryology/growth & development/*microbiology ; Mediterranean Region ; Ovary/microbiology ; Time Factors ; Wolbachia/growth & development/*physiology ; }, abstract = {The Mediterranean flour moth, Ephestia kuehniella, is infected with A-group Wolbachia (wKue), and the almond moth, Cadra cautella, is doubly infected with A- and B-group Wolbachia, which are designated as wCauA and wCauB, respectively. In both insects, the Wolbachia populations increased greatly during embryonic and larval stages. The Wolbachia population doubled every 3.6 days on average in E. kuehniella larvae, whereas those of wCauA and wCauB doubled every 2.1 days in C. cautella larvae. The populations of wCauA and wCauB that had been transferred into the E. kuehniella background increased at similar rates to that of wKue in the natural host E. kuehniella, suggesting that the host genetic background influences Wolbachia proliferation. To examine whether the populations of the two Wolbachia variants in double infection is regulated collectively or independently, we measured the infection load in the ovaries of three transfected E. kuehniella lines in different infection states: single infection with wCauA, single infection with wCauB, and double infection. The density of each Wolbachia variant did not differ significantly between the singly and doubly transfected hosts, suggesting independent regulation.}, } @article {pmid12652910, year = {2002}, author = {Kang, L and Zhu, H and Cheng, Q and Zhou, W and Sun, L and Cai, L and Ma, X and Chen, C and Zhao, S and Li, C}, title = {Cloning and characterization of a gene encoding glutathione-regulated potassium-efflux system protein KefKL from the endosymbiont Wolbachia.}, journal = {DNA sequence : the journal of DNA sequencing and mapping}, volume = {13}, number = {6}, pages = {375-381}, doi = {10.1080/1042517021000003897}, pmid = {12652910}, issn = {1042-5179}, mesh = {Amino Acid Sequence ; Base Sequence ; Glutathione/*metabolism ; Molecular Sequence Data ; Potassium-Hydrogen Antiporters/*genetics/metabolism ; Sequence Analysis, DNA ; Wolbachia/*genetics/metabolism ; }, abstract = {The maternally inherited intracellular symbiont Wolbachia is well known for inducing a variety of reproductive and developmental abnormalities in the diverse arthropod hosts it infects. It has been implicated in causing cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males and male killing in different hosts. However, the molecular mechanisms by which this fastidious bacterium causes these abnormalities have not yet been determined. In our study, representational difference analysis (RDA) was used to analyze the genomic difference between different Wolbachia strains. A gene encoding glutathione-regulated potassium-efflux system protein KefKL from Wolbachia in Drosophila simulans Riverside (w Ri) was isolated. The homologous genes from Wolbachia in Drosophila melanogaster yw67c23 (wMel) and Wolbachia in Drosophila melanogaster CantonS (wMelCS) were also cloned and sequenced. Sequence analysis showed that these deduced amino acid sequences contained two important motifs: Na+/H+ antiportor and NAD binding domain, which shared conserved sequences among different strains. Considering the crucial function of KefKL for ionic homeostasis, this gene might play an important role in Wolbachia physiology. Further study indicated that there was no homologue detected from Wolbachia in Drosophila simulans DSW/Mau (wMa) and Wolbachia in Drosophila simulans Noumea (wNo). Whether Wolbachia contained KefKL (or the homologous gene) was consistent with the phylogenetic studies using wsp sequences, which showed that wMa and wNo were grouped into one branch, while w Ri, wMel and wMelCS were more closely related.}, } @article {pmid12643996, year = {2003}, author = {Dobson, SL}, title = {Reversing Wolbachia-based population replacement.}, journal = {Trends in parasitology}, volume = {19}, number = {3}, pages = {128-133}, doi = {10.1016/s1471-4922(03)00002-3}, pmid = {12643996}, issn = {1471-4922}, mesh = {Animals ; Computer Simulation ; Crosses, Genetic ; Female ; Host-Parasite Interactions ; Infectious Disease Transmission, Vertical ; Infertility/metabolism ; Insecta/parasitology ; Male ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Genetic manipulation that reduces the competence of a vector population to transmit pathogens would provide a useful tool to complement current control strategies, which are based primarily on the reduction/exclusion of vector populations and the prophylactic/therapeutic treatment of the vertebrate host population. Genetic drive is an important component of vector population replacement strategies, facilitating the replacement of natural populations with a genetically modified population. Genetic drive is reviewed here, emphasizing strategies that would employ infections of intracellular Wolbachia bacteria as a vehicle for population replacement. Also discussed are strategies for the retarding, arresting or reversing of Wolbachia-based population replacement. These strategies are based upon altering the conditions required for transgene invasion and are a prudent safeguard, should unexpected detrimental effects become associated with transgene spread.}, } @article {pmid12643829, year = {2003}, author = {Rolain, JM and Franc, M and Davoust, B and Raoult, D}, title = {Molecular detection of Bartonella quintana, B. koehlerae, B. henselae, B. clarridgeiae, Rickettsia felis, and Wolbachia pipientis in cat fleas, France.}, journal = {Emerging infectious diseases}, volume = {9}, number = {3}, pages = {338-342}, pmid = {12643829}, issn = {1080-6040}, mesh = {Animals ; Bartonella/*genetics/isolation & purification ; Cats ; DNA Primers ; France ; Polymerase Chain Reaction ; Rickettsia felis/*genetics/isolation & purification ; Siphonaptera/*microbiology ; Wolbachia/*genetics/isolation & purification ; }, abstract = {The prevalences of Bartonella, Rickettsia, and Wolbachia were investigated in 309 cat fleas from France by polymerase chain reaction (PCR) assay and sequencing with primers derived from the gltA gene for Rickettsia, the its and pap31 genes for Bartonella, and the 16S rRNA gene for Anaplasmataceae. Positive PCR results were confirmed by using the Lightcycler and specific primers for the rOmpB of Rickettsia and gltA of Bartonella. R. felis was detected in 25 fleas (8.1%), W. pipientis, an insect symbiont, in 55 (17.8%), and Bartonella in 81 (26.2%), including B. henselae (9/81; 11.1%), B. clarridgeiae (55/81; 67.9%), B. quintana (14/81; 17.3%), and B. koehlerae (3/81; 3.7%). This is the first report of the amplification of B. quintana from fleas and the first description of B. koehlerae in fleas from an area outside the United States. Cat fleas may be more important vectors of human diseases than previously reported.}, } @article {pmid12634822, year = {2003}, author = {Charlat, S and Bonnavion, P and Merçot, H}, title = {Wolbachia segregation dynamics and levels of cytoplasmic incompatibility in Drosophila sechellia.}, journal = {Heredity}, volume = {90}, number = {2}, pages = {157-161}, doi = {10.1038/sj.hdy.6800211}, pmid = {12634822}, issn = {0018-067X}, mesh = {Animals ; Cytoplasm ; Drosophila/*microbiology/physiology ; Female ; Fertility ; Male ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {In Drosophila sechellia, the endocellular bacterium Wolbachia induces cytoplasmic incompatibility (CI): in crosses involving infected males, a partial or complete embryonic mortality occurs unless the female bears the same Wolbachia. D. sechellia is known to harbour two Wolbachia variants, namely wSh and wSn, closely related to wHa and wNo, respectively, two strains infecting the populations of D. simulans from the Seychelles archipelago and New Caledonia. Strikingly, the two species show similar infection patterns: in D. sechellia, wSh can be present on its own or in double infection with wSn, but individuals carrying wSn only do not occur; in D. simulans, wHa can be present on its own or in double infection with wNo, but individuals carrying wNo only do not occur, or occur at very low frequency. Previous experiments on D. simulans showed that lines singly infected by wNo can be obtained by segregation, and stably maintained. Here we investigate this issue in D. sechellia through an 18 generation experiment, and show that wSn and wSh singly infected lines can arise by segregation. Using singly infected lines obtained in this experiment, we estimate the CI intensities of wSh and wSn in D. sechellia, and compare these to the CI intensities of the same Wolbachia injected into D. simulans. Our results do not suggest any consistent effect of the host species on the CI induced by wSh. On the contrary, it seems that wSn expression is repressed by host factors in D. sechellia.}, } @article {pmid12632152, year = {2003}, author = {Kramer, LH and Passeri, B and Corona, S and Simoncini, L and Casiraghi, M}, title = {Immunohistochemical/immunogold detection and distribution of the endosymbiont Wolbachia of Dirofilaria immitis and Brugia pahangi using a polyclonal antiserum raised against WSP (Wolbachia surface protein).}, journal = {Parasitology research}, volume = {89}, number = {5}, pages = {381-386}, doi = {10.1007/s00436-002-0765-6}, pmid = {12632152}, issn = {0932-0113}, mesh = {Animals ; Antibodies, Bacterial/*immunology ; Antigens, Bacterial/analysis ; Bacterial Outer Membrane Proteins/*analysis/immunology ; Brugia pahangi/*microbiology ; Dirofilaria immitis/*microbiology ; Dirofilariasis/parasitology ; Dogs ; Doxycycline/pharmacology ; Female ; Gerbillinae/parasitology ; Immunohistochemistry ; Male ; Microscopy, Immunoelectron ; Symbiosis ; Wolbachia/drug effects/immunology/*isolation & purification ; }, abstract = {Intracellular bacteria in filarial nematodes were described as early as the 1970s, yet it was only with the work on Dirofilaria immitis, the agent of canine and feline heartworm disease, that these microorganisms were identified as belonging to Wolbachia, a genus known for encompassing bacteria infecting insects and other arthropods. The implications for the presence of intracellular bacteria in filarial nematodes is now the subject of intense research, particularly regarding their role in the immunology and pathogenesis of disease in infected humans and animals and as a possible target for therapy. Here, the authors report results on the immunohistochemical and immunogold staining of Wolbachia in D. immitis and Brugia pahangi using polyclonal antibodies raised against the recombinant Wolbachia surface protein (WSP). The bacteria were present in the lateral hypodermal chords of both male and female worms and in the reproductive tract of adult females (oocytes, morulae, microfilariae). In D. immitis and B. pahangi from animals treated with tetracycline, positive staining was observed in the lateral chords of adult males and females, but was absent from the oocytes and morulae. These results indicate that Wolbachia endosymbionts can be identified immunohistochemically with anti-WSP polyclonal antibodies, that their distribution matches that already described for Wolbachia of other filarial worms, and that antibiotic treatment may impede the vertical transmission of these bacteria. Unequivocal detection of Wolbachia is essential for the study of this symbiont, in particular to monitor the effects of antibiotic treatment on worms. The use of a specific marker for bacteria in their nematode hosts represents an extremely useful tool in evaluating the pathogenic role and the effect of antibiotic treatment on these potential targets in the control of filarial disease.}, } @article {pmid12625936, year = {2002}, author = {Fischer, P and Wibowo, H and Pischke, S and Rückert, P and Liebau, E and Ismid, IS and Supali, T}, title = {PCR-based detection and identification of the filarial parasite Brugia timori from Alor Island, Indonesia.}, journal = {Annals of tropical medicine and parasitology}, volume = {96}, number = {8}, pages = {809-821}, doi = {10.1179/000349802125002239}, pmid = {12625936}, issn = {0003-4983}, mesh = {Animals ; Anopheles/parasitology ; Brugia/*genetics/isolation & purification/microbiology ; Brugia malayi/genetics/isolation & purification ; Brugia pahangi/genetics/isolation & purification ; DNA, Helminth/*analysis ; DNA, Ribosomal Spacer/analysis ; Elephantiasis, Filarial/*parasitology ; Humans ; Indonesia ; Molecular Sequence Data ; Polymerase Chain Reaction/*methods ; Repetitive Sequences, Nucleic Acid ; Sequence Homology, Nucleic Acid ; Wolbachia/genetics ; }, abstract = {Brugia timori is widely distributed on Alor Island, Indonesia, where it causes a high degree of morbidity. The HhaI tandem repeat of B. timori was found to be identical to that of B. malayi, for which sensitive PCR-based assays have already been developed. Using one of these assays, a single microfilaria (mf) of B. timori, present in a spot of dry blood on filter paper, could be detected. The assay was equally sensitive in the detection of B. timori and B. malayi. When the collected mosquitoes were pooled according to species and tested with the assay, 39 (64%) of the 61 Anopheles barbirostris pools (containing a total of 642 mosquitoes) were positive. As none of the 33 Culex pools tested (which contained 624 mosquitoes) gave a positive result, and An. barbirostris is the only Anopheles species commonly caught on human bait in Alor, An. barbirostris is assumed to be the main and perhaps only local vector. Brugia timori could be differentiated from B. malayi by restriction-endonuclease digestion of the PCR-amplified mitochondrial cytochrome oxidase subunit 2. A few distinct nucleotide exchanges were also found in the second internal transcribed ribosomal spacer of the filariae, and in the 16S rDNA and FTSZ gene of their Wolbachia endobacteria. The results show that B. timori can be effectively detected using the PCR-based assay developed for B. malayi and can then be differentiated from B. malayi by other molecular markers. PCR-based techniques targeting the HhaI repeat can therefore be employed for monitoring B. timori in the framework of the Global Programme to Eliminate Lymphatic Filariasis.}, } @article {pmid12625925, year = {2002}, author = {Townson, H}, title = {Wolbachia as a potential tool for suppressing filarial transmission.}, journal = {Annals of tropical medicine and parasitology}, volume = {96 Suppl 2}, number = {}, pages = {S117-27}, doi = {10.1179/000349802125002464}, pmid = {12625925}, issn = {0003-4983}, mesh = {Animals ; Culicidae/genetics/*microbiology/physiology ; Cytoplasm/physiology ; Elephantiasis, Filarial/*prevention & control/transmission ; Gene Transfer Techniques ; Humans ; Pest Control, Biological/*methods ; Population Control/methods ; Reproduction/physiology ; Transgenes/genetics ; Wolbachia/genetics/*physiology/ultrastructure ; }, abstract = {There is currently a great deal of interest in Wolbachia because of their wide distribution in arthropods and filarial nematodes and their striking effects on the biology of their hosts, including a possible role in speciation. They manipulate the reproduction of arthropod hosts through various effects on their hosts' biology, particularly cytoplasmic incompatibility (CI), to increase the proportion of infected individuals in the population, often to the point of fixation. This ability of Wolbachia to sweep through host populations indicates several potential applications of Wolbachia in the control of mosquito-borne disease. One uses Wolbachia-induced CI as a form of sterile-insect technique, to suppress mosquito populations. Another envisages the application of CI for population replacement, with the intention of preventing the transmission of human pathogens, by substituting desirable genotypes, including those carried in transgenes. A third possibility is to use Wolbachia to reduce the survival of mosquito populations and thereby reduce their ability to transmit the infection. This article provides an overview of the biological effects of Wolbachia on arthropod hosts, with discussion of the possible future exploitation of these effects in the control of filariasis.}, } @article {pmid12597651, year = {2003}, author = {Cornel, AJ and McAbee, RD and Rasgon, J and Stanich, MA and Scott, TW and Coetzee, M}, title = {Differences in extent of genetic introgression between sympatric Culex pipiens and Culex quinquefasciatus (Diptera: Culicidae) in California and South Africa.}, journal = {Journal of medical entomology}, volume = {40}, number = {1}, pages = {36-51}, doi = {10.1603/0022-2585-40.1.36}, pmid = {12597651}, issn = {0022-2585}, mesh = {Animals ; Base Sequence ; California ; Culex/*classification/*genetics/microbiology/physiology ; DNA Primers ; Female ; *Genetic Variation ; Genitalia, Male/anatomy & histology ; Geography ; Male ; RNA, Ribosomal, 16S/genetics ; Reproduction ; Sex Ratio ; South Africa ; Wolbachia/pathogenicity ; }, abstract = {Comparisons of five morphological characters, 12 enzyme electrophoresis profiles, and Wolbachia pipientis infection rates were used to characterize populations of members of the Culex pipiens L. complex in California and South Africa. In South Africa, male phallosome DV/D ratio, male maxillary palp index, branching of siphonal seta 1a, the enzyme locus Mdhp-1, and W. pipientis infection rates proved highly diagnostic for separating Culex quinquefasciatus from Cx. pipiens phenotypes. In Johannesburg, where sympatric members of the Cx. pipiens complex were analyzed as one population, a significant Wahlund Effect was observed in the enzyme loci such as Ao, 6-Pgdh, Mdh-2, and Pgm. In California, all populations of the Cx. pipiens complex were in Hardy Weinberg equilibrium at all polymorphic enzyme loci examined. Additionally, in California, all populations had similar W. pipientis infection rates and appeared morphologically identical (except for DV/D ratio, in extreme north and south). These findings indicate that in South Africa, Cx. pipiens and Cx. quinquefasciatus remain as genetically distinct populations and behave as separate species. Conversely, in California, there is considerable genetic introgression between Cx. pipiens and Cx. quinquefasciatus, and they behave as a single species.}, } @article {pmid12597646, year = {2003}, author = {Ruang-Areerate, T and Kittayapong, P and Baimai, V and O'Neill, SL}, title = {Molecular phylogeny of Wolbachia endosymbionts in Southeast Asian mosquitoes (Diptera: Culicidae) based on wsp gene sequences.}, journal = {Journal of medical entomology}, volume = {40}, number = {1}, pages = {1-5}, doi = {10.1603/0022-2585-40.1.1}, pmid = {12597646}, issn = {0022-2585}, mesh = {Aedes/classification/genetics/microbiology ; Animals ; Asia, Southeastern ; Bacterial Outer Membrane Proteins/*genetics ; Base Sequence ; Culicidae/classification/genetics/*microbiology ; DNA Primers ; DNA, Bacterial/*genetics ; Genes, Bacterial ; Phylogeny ; Symbiosis ; Wolbachia/*classification/genetics/physiology ; }, abstract = {Wolbachia are maternally inherited intracellular bacteria that infect a wide range of arthropods and nematodes and are associated with various reproductive abnormalities in their hosts. Insect-associated Wolbachia form a monophyletic clade in the alpha-Proteobacteria and recently have been separated into two supergroups (A and B) and 19 groups. Our recent polymerase chain reaction (PCR) survey using wsp specific primers indicated that various strains of Wolbachia were present in mosquitoes collected from Southeast Asia. Here, we report the phylogenetic relationship of the Wolbachia strains found in these mosquitoes using wsp gene sequences. Our phylogenetic analysis revealed eight new Wolbachia strains, five in the A supergroup and three in the B supergroup. Most of the Wolbachia strains present in Southeast Asian mosquitoes belong to the established Mors, Con, and Pip groups.}, } @article {pmid12596230, year = {2003}, author = {Poinsot, D and Charlat, S and Merçot, H}, title = {On the mechanism of Wolbachia-induced cytoplasmic incompatibility: confronting the models with the facts.}, journal = {BioEssays : news and reviews in molecular, cellular and developmental biology}, volume = {25}, number = {3}, pages = {259-265}, doi = {10.1002/bies.10234}, pmid = {12596230}, issn = {0265-9247}, mesh = {Animals ; Cytoplasm/*metabolism/*microbiology ; Female ; Host-Parasite Interactions ; Male ; Models, Biological ; Ovum/microbiology/physiology ; Reproduction/physiology ; Spermatozoa/*microbiology ; Wolbachia/genetics/*metabolism/*physiology ; }, abstract = {The endocellular bacterium Wolbachia manipulates the reproduction of its arthropod hosts for its own benefit by various means, the most widespread being cytoplasmic incompatibility (CI). To date, the molecular mechanism involved in CI has not been elucidated. We examine here three different CI models described in previous literature, namely, the "lock-and-key", "titration-restitution" and "slow-motion" models. We confront them with the full range of CI patterns discovered so far, including the most complex ones such as multiple infections, asymmetrical and partial compatibility relationships and the existence of Wolbachia variants that can rescue the host from CI but not induce it. We conclude that the lock-and-key model is the most parsimonious of the models and fits the observations best. The two other models cannot be categorically invalidated, but they encounter some difficulties that make additional hypotheses necessary.}, } @article {pmid12592565, year = {2003}, author = {Pearlman, E}, title = {Immunopathogenesis of Onchocerca volvulus keratitis (river blindness): a novel role for endosymbiotic Wolbachia bacteria.}, journal = {Medical microbiology and immunology}, volume = {192}, number = {1}, pages = {57-60}, doi = {10.1007/s00430-002-0160-z}, pmid = {12592565}, issn = {0300-8584}, support = {EY10230/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Disease Models, Animal ; *Drosophila Proteins ; Keratitis/immunology/*microbiology/parasitology ; Membrane Glycoproteins/immunology/metabolism ; Mice ; Mice, Inbred C3H ; Onchocerca volvulus/*immunology ; Onchocerciasis/immunology/*microbiology/parasitology ; Onchocerciasis, Ocular/immunology/*microbiology/parasitology ; Receptors, Cell Surface/immunology/metabolism ; Symbiosis/*immunology ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Wolbachia/*immunology/metabolism ; }, abstract = {River blindness is thought to occur as a result of the host response to degenerating microfilariae in the eye. Utilizing a murine model of corneal inflammation (keratitis) to investigate the immune and inflammatory responses associated with river blindness, we recently demonstrated an important role for endotoxin-like products from endosymbiotic bacteria and for activation of Toll-like receptor 4 (TLR4). These observations have led to a new understanding of the pathogenesis of this disease}, } @article {pmid12592564, year = {2003}, author = {Pfarr, KM and Fischer, K and Hoerauf, A}, title = {Involvement of Toll-like receptor 4 in the embryogenesis of the rodent filaria Litomosoides sigmodontis.}, journal = {Medical microbiology and immunology}, volume = {192}, number = {1}, pages = {53-56}, doi = {10.1007/s00430-002-0159-5}, pmid = {12592564}, issn = {0300-8584}, mesh = {Animals ; Disease Models, Animal ; *Drosophila Proteins ; Female ; Fertility ; Filariasis/parasitology ; Filarioidea/*embryology/physiology ; Membrane Glycoproteins/*physiology ; Mice ; Mice, Inbred C3H ; Rats ; Receptors, Cell Surface/*physiology ; Sigmodontinae ; Toll-Like Receptor 4 ; Toll-Like Receptors ; }, abstract = {To examine the role that lipopolysaccharide (LPS)-like molecules from the filarial intracellular endobacteria Wolbachia might play in the development of filarial infections, a natural infection in the LPS-nonresponsive C3H/HeJ mouse strain was compared to that of the LPS-responsive C3H/HeN mouse strain. C3H/HeN mice have been shown to be susceptible to the rodent filarial nematode Litomosoides sigmodontis, with the development of adult worms including females containing mature microfilariae (first stage larvae) in the uterine tubes. However, free microfilariae are not detected. In this study the worm burden and worm length were not significantly different between the C3H/HeN and C3H/HeJ mice. However, the fertility of worms from CeH/HeJ mice was found to be higher than those from C3H/HeN mice. Significantly, mature microfilariae were found at the site of infection only in C3H/HeJ mice. These results indicate a role for TLR4 signaling in the immune response that inhibits worm embryogenesis and prevents the release of microfilariae or directly kills released microfilariae.}, } @article {pmid12581590, year = {2003}, author = {Marcos-Atxutegi, C and Kramer, LH and Fernandez, I and Simoncini, L and Genchi, M and Prieto, G and Simón, F}, title = {Th1 response in BALB/c mice immunized with Dirofilaria immitis soluble antigens: a possible role for Wolbachia?.}, journal = {Veterinary parasitology}, volume = {112}, number = {1-2}, pages = {117-130}, doi = {10.1016/s0304-4017(02)00419-3}, pmid = {12581590}, issn = {0304-4017}, mesh = {Animals ; Antibodies, Bacterial/immunology ; Antibodies, Helminth/immunology ; Antigens, Helminth/chemistry/*immunology ; Cytokines/blood ; Dirofilaria immitis/chemistry/*immunology/*microbiology ; Eosinophils/immunology ; Female ; Immunoglobulin E/blood ; Immunoglobulin G/blood ; Leukocyte Count ; Mice ; Mice, Inbred BALB C ; Neutrophils/immunology ; Solubility ; Th1 Cells/*immunology ; Th2 Cells/immunology ; Time Factors ; Wolbachia/*immunology/*physiology ; }, abstract = {The immune response to filarial infection has been shown to be of both the Th1 and Th2 types. Studies aimed at developing immunization strategies against Dirofilaria immitis infection in dogs have shown that protection against larval challenge is of the Th2 type and that several proteins are recognized by immunized or infected animals. The bacterial endosymbiont Wolbachia, harbored by many filarial species including D. immitis, has recently been shown to interact with the host immune system. Specific antibodies to the Wolbachia recombinant surface protein (WSPr) have been observed in cats infected with D. immitis. In this work the authors have determined cytokine production and antibody response in BALB/c mice inoculated with soluble antigens from third stage larvae or from adult worms of D. immitis. Inoculated mice first produced IFN-gamma followed by a peak in IL-4. Specific antibodies to the Wolbachia protein WSPr were exclusively IgG2a, while antibodies against peptides derived from antigens of D. immitis were in the IgG1 and IgE subclasses. The cytokine response is thus similar to that reported for other filarial infection, where Th1 response shifts towards Th2. Antibody response indicates that Wolbachia may induce preferentially a Th1 response during filarial infection, while nematode antigens may be involved in Th2 response. There is thus an overall agreement with current opinions on the role of bacterial versus nematode molecules in driving the response towards the different directions.}, } @article {pmid12573064, year = {2002}, author = {Groenenboom, MA and Hogeweg, P}, title = {Space and the persistence of male-killing endosymbionts in insect populations.}, journal = {Proceedings. Biological sciences}, volume = {269}, number = {1509}, pages = {2509-2518}, pmid = {12573064}, issn = {0962-8452}, mesh = {Animals ; Female ; Insecta/growth & development/*microbiology ; Male ; Models, Biological ; Polymorphism, Genetic ; Sex Ratio ; *Symbiosis ; Wolbachia/genetics/*growth & development/*pathogenicity ; }, abstract = {Male-killing bacteria are bacteria that are transmitted vertically through the females of their insect hosts. They can distort the sex ratio of their hosts by killing infected male offspring. In nature, male-killing endosymbionts (male killers) often have a 100% efficient vertical transmission, and multiple male-killing bacteria infecting a single population are observed. We use different model formalisms to study these observations. In mean-field models a male killer with perfect transmission drives the host population to extinction, and coexistence between multiple male killers within one population is impossible; however, in spatially explicit models, both phenomena are readily observed. We show how the spatial pattern formation underlies these results. In the case of high transmission efficiencies, waves with a high density of male killers alternate with waves of mainly wild-type hosts. The male killers cause local extinction, but this creates an opportunity for uninfected hosts to re-invade these areas. Spatial pattern formation also creates an opportunity for two male killers to coexist within one population: different strains create spatial regions that are qualitatively different; these areas then serve as different niches, making coexistence possible.}, } @article {pmid12559491, year = {2003}, author = {Clark, ME and Veneti, Z and Bourtzis, K and Karr, TL}, title = {Wolbachia distribution and cytoplasmic incompatibility during sperm development: the cyst as the basic cellular unit of CI expression.}, journal = {Mechanisms of development}, volume = {120}, number = {2}, pages = {185-198}, doi = {10.1016/s0925-4773(02)00424-0}, pmid = {12559491}, issn = {0925-4773}, mesh = {Animals ; Cysts ; Cytoplasm/*physiology ; Drosophila/genetics/*microbiology ; Drosophila melanogaster/genetics/microbiology ; Female ; Fertility/genetics/physiology ; Host-Parasite Interactions/*physiology ; Male ; Spermatocytes/microbiology ; Spermatogenesis/*physiology ; Spermatozoa/microbiology ; Testis/microbiology ; Wolbachia/genetics/*physiology ; }, abstract = {The growth and distribution of the intracellular microbe Wolbachia pipientis during spermatogenesis in several different host/symbiont genetic combinations in Drosophila melanogaster and Drosophila simulans is described. Considerable intra- and inter-strain variation in Wolbachia density and tissue distribution was observed. Wolbachia were found inside spermatocytes and spermatids or within the somatic cyst cells surrounding the germ cells. Some strains displayed both tissue distributions. High rates of cytoplasmic incompatibility (CI) are correlated with high levels of Wolbachia only when spermatocytes and/or spermatids harbor the microbe. Wolbachia infection of somatic cyst cells, although sometimes present at high levels, did not result in significant CI expression. CI-inducing Wolbachia strains within D. simulans showed no distinguishable differences in distribution or density within infected spermatids. To dissect the relative contribution of host and symbiont to the expression of CI, Wolbachia from various host strains known to exhibit varying levels of CI were introgressed into new uninfected host genetic backgrounds. These introgression experiments confirm that the mod(+)/mod(-) phenotype is an intrinsic Wolbachia trait and is not determined by host factors. The level of sperm modification in those lines harboring Wolbachia capable of modifying sperm, however, is influenced by host genetic background. These results form the basis of the Wolbachia Infected Spermatocyte/Spermatid Hypothesis (WISSH). According to WISSH, Wolbachia infection in spermatocytes and then spermatids during sperm development is required for CI expression.}, } @article {pmid12546803, year = {2003}, author = {Gogarten, JP}, title = {Gene transfer: gene swapping craze reaches eukaryotes.}, journal = {Current biology : CB}, volume = {13}, number = {2}, pages = {R53-4}, doi = {10.1016/s0960-9822(02)01426-4}, pmid = {12546803}, issn = {0960-9822}, mesh = {Animals ; *Biological Evolution ; Diplomonadida/genetics ; Eukaryotic Cells ; *Gene Transfer, Horizontal ; Genes, Bacterial ; Genes, Protozoan ; Hybridization, Genetic ; Models, Genetic ; Phagocytosis ; Prokaryotic Cells ; Symbiosis ; Wolbachia/genetics ; }, abstract = {Recent studies have provided evidence for gene transfers from prokaryotes to eukaryotes and between eukaryotes. The mechanisms and frequencies of these transfers remain the subject of speculation, but the findings provide ample reason to seriously consider interspecies gene transfer as an important evolutionary process in eukaryotes.}, } @article {pmid12543839, year = {2003}, author = {Hoerauf, A and Büttner, DW and Adjei, O and Pearlman, E}, title = {Onchocerciasis.}, journal = {BMJ (Clinical research ed.)}, volume = {326}, number = {7382}, pages = {207-210}, pmid = {12543839}, issn = {1756-1833}, support = {//Wellcome Trust/United Kingdom ; R01 EY010320/EY/NEI NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Doxycycline/therapeutic use ; Forecasting ; Humans ; Onchocerca volvulus/microbiology ; Onchocerciasis/microbiology/*prevention & control ; Rickettsiaceae Infections/*microbiology/prevention & control ; *Wolbachia ; }, } @article {pmid12543723, year = {2002}, author = {Lammie, PJ and Cuenco, KT and Punkosdy, GA}, title = {The pathogenesis of filarial lymphedema: is it the worm or is it the host?.}, journal = {Annals of the New York Academy of Sciences}, volume = {979}, number = {}, pages = {131-42; discussion 188-96}, doi = {10.1111/j.1749-6632.2002.tb04874.x}, pmid = {12543723}, issn = {0077-8923}, mesh = {Animals ; Elephantiasis, Filarial/immunology/*physiopathology ; Haiti ; *Host-Parasite Interactions ; Humans ; Lymphedema/*parasitology ; Wuchereria bancrofti ; }, abstract = {Our understanding of the pathogenesis of filarial lymphedema, although evolving, is still limited. Recurrent bacterial infections play a major role in the progression of lymphedema to elephantiasis, but the host and parasite factors that trigger disease development are not known. Field studies in Haiti show that lymphedema and host responses to parasite antigens cluster in families, consistent with the hypothesis that host genes influence lymphedema susceptibility. The recent recognition that filarial parasites harbor the endosymbiotic bacteria, Wolbachia, also raises questions about the potential contribution of the inflammatory response to Wolbachia antigens to lymphedema development. In this review, we discuss potential risk factors for lymphedema and try to integrate these in a model of pathogenesis.}, } @article {pmid12537121, year = {2002}, author = {Chirgwin, SR and Porthouse, KH and Nowling, JM and Klei, TR}, title = {The filarial endosymbiont Wolbachia sp. is absent from Setaria equina.}, journal = {The Journal of parasitology}, volume = {88}, number = {6}, pages = {1248-1250}, doi = {10.1645/0022-3395(2002)088[1248:TFEWSI]2.0.CO;2}, pmid = {12537121}, issn = {0022-3395}, support = {AI-19199-16/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Female ; Horse Diseases/parasitology ; Horses ; Male ; Peritoneal Cavity/parasitology ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics ; Setaria Nematode/microbiology/*physiology ; Setariasis/parasitology ; *Symbiosis ; Wolbachia/genetics/isolation & purification/*physiology ; }, abstract = {Wolbachia sp. was first reported in filarial nematodes over 25 yr ago. Today, much research is focused on the role of these bacteria in filarial worm biology. The filarial symbionts are closely related to arthropod symbionts, which are known to modify host reproduction and biology through various mechanisms. Similarly, it has been suggested that Wolbachia sp. is essential for long-term survival and reproduction of filariae. We report that Wolbachia sp. 16S rDNA was not found in the equine filarial nematode Setaria equina, using either polymerase chain reaction (PCR) or DNA hybridization. In addition, ultrastructural analysis of adult worms did not reveal the presence of Wolbachia sp. in hypodermal cords or reproductive tissues. These data suggest that like Onchocerca flexuosa and Acanthocheilonema vitae, S. equina may not be dependent on Wolbachia sp. for survival.}, } @article {pmid12534342, year = {2002}, author = {Noda, H and Miyoshi, T and Koizumi, Y}, title = {In vitro cultivation of Wolbachia in insect and mammalian cell lines.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {38}, number = {7}, pages = {423-427}, pmid = {12534342}, issn = {1071-2690}, mesh = {Aedes/cytology/microbiology ; Animals ; Cell Line ; Insecta/*cytology/*microbiology ; Lepidoptera/cytology/microbiology ; Mice ; Time Factors ; Wolbachia/genetics/*growth & development/ultrastructure ; }, abstract = {Wolbachia infecting the small brown planthopper, Laodelphax striatellus, were successfully maintained and cultivated in two insect and one mammalian cell lines. The bacteria with the planthopper ovary were introduced into the flasks with the cultures of the cell lines. The Wolbachia proliferated in mosquito (Aedes albopictus) and lepidopteran (Heliothis zea) cell lines and in the mouse cell line, L929. Proliferation of Wolbachia was confirmed by electron microscopy and quantitative polymerase chain reaction. This simple method for the cultivation of Wolbachia was applicable to other strains of Wolbachia, such as the one found in the lepidopteran eggs, and should facilitate fundamental and applied studies of this important group of microorganisms.}, } @article {pmid12534211, year = {2002}, author = {Reynolds, KT and Hoffmann, AA}, title = {Male age, host effects and the weak expression or non-expression of cytoplasmic incompatibility in Drosophila strains infected by maternally transmitted Wolbachia.}, journal = {Genetical research}, volume = {80}, number = {2}, pages = {79-87}, doi = {10.1017/s0016672302005827}, pmid = {12534211}, mesh = {Age Factors ; Animals ; Crosses, Genetic ; Cytoplasm/*physiology ; DNA, Bacterial/genetics ; Drosophila/*microbiology ; Female ; Male ; *Wolbachia ; }, abstract = {In Drosophila melanogaster, the maternally inherited endocellular microbe Wolbachia causes cytoplasmic incompatibility (CI) in crosses between infected males and uninfected females. CI results in a reduction in the number of eggs that hatch. The level of CI expression in this species has been reported as varying from partial (a few eggs fail to hatch) to nonexistent (all eggs hatch). We show that male age in this host species has a large impact on the level of CI exhibited and explains much of this variability. Strong CI is apparent when young males are used in crosses. CI declines rapidly with male age, particularly when males are repeatedly mated. Wolbachia from a Canton S line that was previously reported as not causing CI does in fact induce CI when young males are used in crosses, albeit at a weaker level than in other D. melanogaster strains. The strain differences in CI expression are due to host background effects rather than differences in Wolbachia strains. These results highlight the importance of undertaking crosses with a range of male ages and nuclear backgrounds before ascribing particular host phenotypes to Wolbachia strains.}, } @article {pmid12522428, year = {2003}, author = {Kang, L and Ma, X and Cai, L and Liao, S and Sun, L and Zhu, H and Chen, X and Shen, D and Zhao, S and Li, C}, title = {Superinfection of Laodelphax striatellus with Wolbachia from Drosophila simulans.}, journal = {Heredity}, volume = {90}, number = {1}, pages = {71-76}, doi = {10.1038/sj.hdy.6800180}, pmid = {12522428}, issn = {0018-067X}, mesh = {Animals ; Drosophila/genetics/*microbiology ; Insect Vectors/genetics/virology ; Oryza/genetics/virology ; *Superinfection/genetics/transmission ; Tenuivirus/genetics ; Wolbachia/*genetics ; }, abstract = {Wolbachia are maternally inherited, intracellular alpha-proteobacteria that infect a wide range of arthropods. They manipulate the reproduction of hosts to facilitate their spread into host populations, through ways such as cytoplasmic incompatibility (CI), parthenogenesis, feminization and male killing. The influence of Wolbachia infection on host populations has attracted considerable interest in their possible role in speciation and as a potential agent of biological control. In this study, we used both microinjection and nested PCR to show that the Wolbachia naturally infecting Drosophila simulans can be transferred into a naturally Wolbachia-infected strain of the small brown planthopper Laodelphax striatellus, with up to 30% superinfection frequency in the F(12) generation. The superinfected males of L. striatellus showed unidirectional CI when mated with the original single-infected females, while superinfected females of L. striatellus were compatible with superinfected or single-infected males. These results are, to our knowledge, the first to establish a superinfected horizontal transfer route for Wolbachia between phylogenetically distant insects. The segregation of Wolbachia from superinfected L. striatellus was observed during the spreading process, which suggests that Wolbachia could adapt to a phylogenetically distant host with increased infection frequency in the new host population; however, it would take a long time to establish a high-frequency superinfection line. This study implies a novel way to generate insect lines capable of driving desired genes into Wolbachia-infected populations to start population replacement.}, } @article {pmid12522425, year = {2003}, author = {Charlat, S and Le Chat, L and Merçot, H}, title = {Characterization of non-cytoplasmic incompatibility inducing Wolbachia in two continental African populations of Drosophila simulans.}, journal = {Heredity}, volume = {90}, number = {1}, pages = {49-55}, doi = {10.1038/sj.hdy.6800177}, pmid = {12522425}, issn = {0018-067X}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; DNA, Mitochondrial ; Drosophila/*microbiology ; Haplotypes ; Phylogeny ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an endocellular bacterium infecting arthropods and nematodes. In arthropods, it invades host populations through various mechanisms, affecting host reproduction, the most common of which being cytoplasmic incompatibility (CI). CI is an embryonic mortality occurring when infected males mate with uninfected females or females infected by a different Wolbachia strain. This phenomenon is observed in Drosophila simulans, an intensively studied Wolbachia host, harbouring at least five distinct bacterial strains. In this study, we investigate various aspects of the Wolbachia infections occurring in two continental African populations of D. simulans: CI phenotype, phylogenetic position based on the wsp gene and associated mitochondrial haplotype. From the East African population (Tanzania), we show that (i) the siIII mitochondrial haplotype occurs in continental populations, which was unexpected based on the current views of D. simulans biogeography, (ii) the wKi strain (that rescues from CI while being unable to induce it) is very closely related to the CI-inducing strain wNo, (iii) wKi and wNo might not derive from a unique infection event, and (iv) wKi is likely to represent the same entity as the previously described wMa variant. In the West African population (Cameroon), the Wolbachia infection was found identical to the previously described wAu, which does not induce CI. This finding supports the view that wAu might be an ancient infection in D. simulans.}, } @article {pmid12522059, year = {2003}, author = {Simón, F and Prieto, G and Morchón, R and Bazzocchi, C and Bandi, C and Genchi, C}, title = {Immunoglobulin G antibodies against the endosymbionts of filarial nematodes (Wolbachia) in patients with pulmonary dirofilariasis.}, journal = {Clinical and diagnostic laboratory immunology}, volume = {10}, number = {1}, pages = {180-181}, pmid = {12522059}, issn = {1071-412X}, mesh = {Animals ; Antibodies, Bacterial/*blood ; Antibody Formation ; Dirofilaria ; Dirofilariasis/*diagnosis/epidemiology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G/blood ; Solitary Pulmonary Nodule/parasitology ; Symbiosis ; Wolbachia/immunology ; }, abstract = {The dog parasite Dirofilaria immitis can infect humans. Patients with pulmonary dirofilariasis were tested for immunoglobulin G (IgG) antibodies against the surface protein of Wolbachia, the bacterial endosymbiont of D. immitis. These patients showed significantly higher IgG titers than healthy individuals from areas in which D. immitis was endemic as well as areas in which it was not endemic. Titration of anti-Wolbachia surface protein IgG could become useful for diagnostic applications.}, } @article {pmid12507486, year = {2002}, author = {Gottlieb, Y and Zchori-Fein, E and Werren, JH and Karr, TL}, title = {Diploidy restoration in Wolbachia-infected Muscidifurax uniraptor (Hymenoptera: Pteromalidae).}, journal = {Journal of invertebrate pathology}, volume = {81}, number = {3}, pages = {166-174}, doi = {10.1016/s0022-2011(02)00149-0}, pmid = {12507486}, issn = {0022-2011}, mesh = {Animals ; Bacterial Infections/*genetics ; *Diploidy ; Female ; Hymenoptera/*genetics/*microbiology ; *Wolbachia ; }, abstract = {Thelytokous reproduction, where females produce diploid female offspring without fertilization, can be found in many insects. In some Hymenoptera species, thelytoky is induced by Wolbachia, a group of cytoplasmically inherited bacteria. We compare and contrast early embryonic development in the thelytokous parthenogenetic species Muscidifurax uniraptor with the development of unfertilized eggs of the closely related arrhenotokous species, Muscidifurax raptorellus. In the Wolbachia-infected parasitic wasp M. uniraptor, meiosis and the first mitotic division occur normally. Diploidy restoration is achieved following the completion of the first mitosis. This pattern differs in the timing of diploidy restoration from previously described cases of Wolbachia-associated thelytoky. Results presented here suggest that different cytogenetic mechanisms of diploidy restoration may occur in different species with Wolbachia-induced thelytoky.}, } @article {pmid12487358, year = {2002}, author = {Jiggins, FM and Randerson, JP and Hurst, GD and Majerus, ME}, title = {How can sex ratio distorters reach extreme prevalences? Male-killing Wolbachia are not suppressed and have near-perfect vertical transmission efficiency in Acraea encedon.}, journal = {Evolution; international journal of organic evolution}, volume = {56}, number = {11}, pages = {2290-2295}, doi = {10.1111/j.0014-3820.2002.tb00152.x}, pmid = {12487358}, issn = {0014-3820}, mesh = {Animals ; Butterflies/*microbiology/physiology ; Female ; Male ; *Sex Ratio ; Sexual Behavior, Animal/physiology ; Wolbachia/*physiology ; }, abstract = {Maternally transmitted bacteria that kill male hosts early in their development are found in many insects. These parasites typically infect 1-30% of wild females, but in a few species of insects, prevalences exceed 95%. We investigated one such case in the butterfly Acraea encedon, which is infected with a male-killing Wolbachia bacterium. We measured three key parameters that affect the prevalence of the parasite: transmission efficiency, rate of survival of infected males, and the direct cost of infection. We observed that all wild females transmit the bacterium to all their offspring and that all infected males die in wild populations. We were unable to detect any physiological cost to infection in lab culture. These observations explain the high prevalence of the A. encedon male killer, as theory predicts that under these conditions the parasite will spread to fixation. This will occur provided the death of males provides some benefit to the surviving infected females. The problem therefore becomes why the bacterium has not reached fixation and driven the butterfly extinct due to the shortage of males. We therefore investigated whether males choose to mate with uninfected rather than infected females, as this would prevent the bacterium from reaching fixation. We tested this hypothesis in the "lekking swarms" of virgin females found in the most female-biased populations, and were unable to detect any evidence of mate choice. In conclusion, this male killer has spread to high prevalence because it has a high transmission efficiency and low cost, but the factors maintaining uninfected females in the population remain unknown.}, } @article {pmid12470942, year = {2003}, author = {Van Borm, S and Wenseleers, T and Billen, J and Boomsma, JJ}, title = {Cloning and sequencing of wsp encoding gene fragments reveals a diversity of co-infecting Wolbachia strains in Acromyrmex leafcutter ants.}, journal = {Molecular phylogenetics and evolution}, volume = {26}, number = {1}, pages = {102-109}, doi = {10.1016/s1055-7903(02)00298-1}, pmid = {12470942}, issn = {1055-7903}, mesh = {Animals ; Ants/*microbiology ; Bacterial Outer Membrane Proteins/*genetics ; *Cloning, Molecular ; Phylogeny ; Sequence Analysis, DNA ; Wolbachia/*genetics/metabolism ; }, abstract = {By sequencing part of the wsp gene of a series of clones, we detected an unusually high diversity of nine Wolbachia strains in queens of three species of leafcutter ants. Up to four strains co-occurred in a single ant. Most strains occurred in two clusters (InvA and InvB), but the social parasite Acromyrmex insinuator hosted two additional infections. The multiple Wolbachia strains may influence the expression of reproductive conflicts in leafcutter ants, but the expected turnover of infections may make the cumulative effects on host ant reproduction complex. The additional Wolbachia infections of the social parasite A. insinuator were almost certainly acquired by horizontal transmission, but may have facilitated reproductive isolation from its closely related host.}, } @article {pmid12454075, year = {2002}, author = {Sasaki, T and Kubo, T and Ishikawa, H}, title = {Interspecific transfer of Wolbachia between two lepidopteran insects expressing cytoplasmic incompatibility: a Wolbachia variant naturally infecting Cadra cautella causes male killing in Ephestia kuehniella.}, journal = {Genetics}, volume = {162}, number = {3}, pages = {1313-1319}, pmid = {12454075}, issn = {0016-6731}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Gram-Negative Bacterial Infections/physiopathology ; Moths/drug effects/*microbiology ; Phenotype ; Sex Ratio ; Tetracycline/pharmacology ; Transfection ; Wolbachia/drug effects/*pathogenicity ; }, abstract = {Wolbachia is known as the causative agent of various reproductive alterations in arthropods. The almond moth Cadra cautella is doubly infected with A- and B-group Wolbachia and expresses complete cytoplasmic incompatibility (CI). The Mediterranean flour moth Ephestia kuehniella carries A-group Wolbachia and expresses partial CI. In the present study, the Wolbachia in C. cautella was transferred to E. kuehniella from which the original Wolbachia had been removed. We obtained transfected lines of three different infection states: single infection with A, single infection with B, and double infection with A and B. The doubly transfected lines and those transfected with only A produced exclusively female progeny. Two lines of evidence suggested that the sex ratio distortion was due to male killing. First, reduced egg hatch rate was observed. Second, removal of the Wolbachia from the transfected lines resulted in the recovery of a normal sex ratio of approximately 1:1. The occurrence of male killing following transfection showed that host factors influence the determination of the reproductive phenotype caused by Wolbachia. The transfected E. kuehniella males carrying exclusively B-group Wolbachia expressed partial incompatibility when crossed with the uninfected females. In addition, the transfected lines were bidirectionally incompatible with the naturally infected strain, which was the first demonstration of bidirectional CI in a lepidopteran.}, } @article {pmid12449484, year = {2002}, author = {Fry, AJ and Rand, DM}, title = {Wolbachia interactions that determine Drosophila melanogaster survival.}, journal = {Evolution; international journal of organic evolution}, volume = {56}, number = {10}, pages = {1976-1981}, doi = {10.1111/j.0014-3820.2002.tb00123.x}, pmid = {12449484}, issn = {0014-3820}, mesh = {Animals ; Crosses, Genetic ; Drosophila melanogaster/*microbiology ; Female ; Fertility ; Genotype ; Longevity ; Male ; RNA, Ribosomal, 16S/metabolism ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*pathogenicity/*physiology ; }, abstract = {We have recently described a mutualistic symbiosis in which Wolbachia bacteria were shown to improve the fitness of some Drosophila melanogaster stocks. Wolbachia did not extend longevity in all Drosophila genotypes, even though 16s rDNA sequences indicated that our Drosophila stocks were infected with the same Wolbachia strain. Here, we use reciprocal hybrid crosses between two Drosophila strains, one that lived longer with Wolbachia (Z53) and one that did not (Z2), to investigate the inheritance of the survival phenotype and its dependence on the host genotype, sex, and mating conditions. Wolbachia's positive effects were more apparent in hybrid flies than in parental flies, ruling out exclusive maternal inheritance or the dependence of the survival phenotype on Wolbachia strain differences. The Wolbachia survival effects were more apparent in single-sex cages, where courtship and mating were not permitted. In these cages, nearly all flies with Wolbachia lived longer than uninfected flies, even though strain Z2 showed no Wolbachia effect in mixed-sex mating cages. We used comparisons between single- and mixed-sex cages to estimate the cost of reproduction for both sexes. Our data suggest that Wolbachia infection may increase the inferred cost of reproduction, particularly in males. Wolbachia can even produce a positive survival effect almost as large as the negative survival effect associated with reproduction. We discuss the implications of our experiments for the study of insect symbioses.}, } @article {pmid12440772, year = {2002}, author = {Vray, B and Hartmann, S and Hoebeke, J}, title = {Immunomodulatory properties of cystatins.}, journal = {Cellular and molecular life sciences : CMLS}, volume = {59}, number = {9}, pages = {1503-1512}, doi = {10.1007/s00018-002-8525-4}, pmid = {12440772}, issn = {1420-682X}, mesh = {Amino Acid Sequence ; Animals ; Cathepsins/metabolism ; Cystatins/genetics/*immunology/metabolism ; Cysteine Proteinase Inhibitors/genetics/*immunology/metabolism ; Cytokines/metabolism ; Helminth Proteins/genetics/immunology/metabolism ; Humans ; Models, Molecular ; Molecular Sequence Data ; Nitric Oxide/metabolism ; Protein Structure, Tertiary ; Sequence Alignment ; T-Lymphocyte Subsets/immunology/metabolism ; Wolbachia/metabolism ; }, abstract = {Cystatins are natural tight-binding reversible inhibitors of cysteine proteases. Because these cysteine proteases exist in all living organisms and because they are involved in various biological and pathological processes, the control of these protease functions by cystatins is of cardinal importance. Cystatins are found in mammals but cystatin-like molecules are also present in mammals and parasites. In the immune system, cystatins modulate cathepsin activities and antigen presentation. They also induce tumor necrosis factor alpha and interleukin 10 synthesis, and they stimulate nitric oxide production by interferon gamma-activated murine macrophages. In turn, nitric oxide has inhibitory activity on cysteine proteases, especially those from parasitic protozoa. Cystatins isolated from parasitic nematodes also have immunomodulatory activities that are distinguishable from those induced by lipopolysacharide-like molecules from endosymbiotic bacteria. On the whole, cystatins and cystatin-like molecules belong to a new category of immunomodulatory molecules. Doubtless increasing data will improve our knowledge of this property, leading to practical applications in immunotherapy.}, } @article {pmid12427319, year = {2002}, author = {Shoemaker, DD and Machado, CA and Molbo, D and Werren, JH and Windsor, DM and Herre, EA}, title = {The distribution of Wolbachia in fig wasps: correlations with host phylogeny, ecology and population structure.}, journal = {Proceedings. Biological sciences}, volume = {269}, number = {1506}, pages = {2257-2267}, pmid = {12427319}, issn = {0962-8452}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; DNA, Bacterial/genetics ; Ecosystem ; Female ; Ficus/parasitology ; Genes, Bacterial ; Male ; Panama ; Phylogeny ; Population Dynamics ; Symbiosis ; Wasps/genetics/*microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {We surveyed for the presence and identity of Wolbachia in 44 species of chalcid wasps associated with 18 species of Panamanian figs. We used existing detailed knowledge of the population structures of the host wasps, as well as the ecological and evolutionary relationships among them, to explore the relevance of each of these factors to Wolbachia prevalence and mode of transmission. Fifty-nine per cent of these wasp species have Wolbachia infections, the highest proportion reported for any group of insects. Further, neither the presence nor the frequency of Wolbachia within hosts was correlated with the population structure of pollinator hosts. Phylogenetic analyses of wsp sequence data from 70 individuals representing 22 wasp species show that neither the close phylogenetic relationship nor close ecological association among host species is consistently linked to close phylogenetic affinities of the Wolbachia associated with them. Moreover, no genetic variation was detected within any Wolbachia strain from a given host species. Thus, the spread of Wolbachia within host species exceeds the rate of horizontal transmission among species and both exceed the rate of mutation of the wsp gene in Wolbachia. The presence and, in some cases, high frequency of Wolbachia infections within highly inbred species indicate that the Wolbachia either directly increase host fitness or are frequently horizontally transferred within these wasp species. However, the paucity of cospeciation of Wolbachia and their wasp hosts indicates that Wolbachia do not persist within a given host lineage for long time-periods relative to speciation times.}, } @article {pmid12421415, year = {2002}, author = {Mitsuhashi, W and Saiki, T and Wei, W and Kawakita, H and Sato, M}, title = {Two novel strains of Wolbachia coexisting in both species of mulberry leafhoppers.}, journal = {Insect molecular biology}, volume = {11}, number = {6}, pages = {577-584}, doi = {10.1046/j.1365-2583.2002.00368.x}, pmid = {12421415}, issn = {0962-1075}, mesh = {Animals ; DNA Primers ; DNA, Ribosomal/genetics ; Female ; Fruit/parasitology ; Insecta/*microbiology ; Ovary/microbiology ; Ovum/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Salivary Glands/microbiology ; Trees/parasitology ; Wolbachia/*classification/genetics/isolation & purification ; }, abstract = {Wolbachia is an intracellular symbiont that causes reproductive disorders in many insects. Its presence in the leafhoppers Hishimonoides sellatiformis and Hishimonus sellatus, vectors of mulberry dwarf-Phytoplasma, was confirmed by the PCR analysis of 16S rDNA, ftsZ and wsp. Sequencing of cloned PCR products revealed that two Wolbachia strains coexist in both leafhoppers. The phylogenetic analysis of wsp revealed that these strains belong in novel positions in the B-group of Wolbachia. These strains were detected by PCR and/or PCR-RFLP in all of the tested non-genital organs including salivary glands, as well as in the tested genital organs of Hishimonoides sellatiformis. In addition, Wolbachia-like organisms were observed by electron microscopy in all PCR-positive organs. We discuss the possible horizontal transmission of Wolbachia via mulberry trees.}, } @article {pmid12406252, year = {2002}, author = {Riegler, M and Stauffer, C}, title = {Wolbachia infections and superinfections in cytoplasmically incompatible populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae).}, journal = {Molecular ecology}, volume = {11}, number = {11}, pages = {2425-2434}, doi = {10.1046/j.1365-294x.2002.01614.x}, pmid = {12406252}, issn = {0962-1083}, mesh = {Animals ; Cytoplasm/genetics ; Europe ; Genetics, Population ; Host-Parasite Interactions ; Infertility/microbiology ; Molecular Sequence Data ; Species Specificity ; Tephritidae/classification/*microbiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia is an obligately intracellular, maternally inherited bacterium which has been detected in many arthropods. Wolbachia infections disperse in host populations by mechanisms such as cytoplasmic incompatibility (CI). CI leads to embryonic mortality which occurs when infected males mate with uninfected females or females with a different Wolbachia strain. Populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae) were found to be infected by two different Wolbachia strains, wCer1 and wCer2. Superinfections with both strains occurred throughout southern and central Europe and infections with wCer1 were found in northern, western and eastern Europe. Strong unidirectional CI between European populations of R. cerasi were first reported in the 1970s. From the conformity in the recent geographical distribution of the Wolbachia infections and the CI expression patterns found 25 years ago it was deduced that wCer2 potentially causes CI in R. cerasi. The comparison of the geographical distributions indicated that wCer1 + 2 must have spread into wCer1-infected populations in some areas. In other regions, a spread of wCer1 + 2 was probably prevented by dispersal barriers. There, a sharp transition from infected to superinfected populations suggested regional isolation between wCer1 and wCer1 + 2-infected populations.}, } @article {pmid12403324, year = {2002}, author = {Kläger, SL and Watson, A and Achukwi, D and Hultmark, D and Hagen, HE}, title = {Humoral immune response of Simulium damnosum s.l. following filarial and bacterial infections.}, journal = {Parasitology}, volume = {125}, number = {Pt 4}, pages = {359-366}, doi = {10.1017/s0031182002002159}, pmid = {12403324}, issn = {0031-1820}, mesh = {Animals ; Antibody Formation ; Antimicrobial Cationic Peptides/biosynthesis/immunology ; Bacterial Infections/*immunology/microbiology ; Cattle ; Cell Extracts/immunology ; Cell Line ; Drosophila melanogaster ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli/immunology ; Female ; Hemolymph/chemistry/immunology ; Lipopolysaccharides/*immunology ; Micrococcus luteus/immunology ; Onchocerca/*immunology/microbiology ; Onchocerciasis/*immunology/microbiology ; Peptides/blood/immunology ; Simuliidae/*immunology/microbiology/parasitology ; Time Factors ; Wolbachia/immunology ; }, abstract = {The time-course of the humoral immune response of female blackflies after a challenge with bacteria, different Onchocerca microfilariae species, bacterial endotoxin and microfilarial extract was investigated. Strong bacteriolytic and growth inhibition activities against the Gram-positive bacterium Micrococcus luteus were induced by all agents. Specific differences were found in activity levels and time-course. Notably the endotoxin lipopolysaccharide (LPS) induced a very early, profound bacteriolytic and antibacterial response, which declined within a day after injection. In contrast, the bacteriolytic activities after Escherichia coli D31 and Onchocerca microfilariae infections were lower, but remained elevated over the observation period of 4 days. The bacteriolytic activity was correlated to a haemolymph protein with a molecular weight of around 14 kDa. Anti-Gram-positive activity in the E. coli infected group appeared within the first 6 h. However, it took 4 days in the microfilarial infected blackflies to reach significant levels. The active agent was identified to be a peptide with a molecular weight of around 4-4.5 kDa. Activity against the Gram-negative bacteria E. coli was detected in blackflies injected with E. coli D31, O. dukei microfilariae and microfilarial extract on days 1 and 4 after injection. The immune response in S. damnosum s.l. naturally infected via a bloodmeal on cattle supported the findings of the experimental infections. Similarities of the immune response kinetics between bacterial and filarial infections suggested that intracellular Wolbachia bacteria, released from microfilariae, could be responsible for the antibacterial response. This is supported by the observation that the induction of an immune response in the Drosophila melanogaster mbn-2 cell line by the filarial extract is blocked by polymyxin B, which forms inactive complexes with bacterial LPS.}, } @article {pmid12402526, year = {2001}, author = {Simoncini, L and Casiraghi, M and Bazzocchi, C and Sacchi, L and Bandi, C and Genchi, C}, title = {Real-time PCR for quantification of the bacterial endosymbionts (Wolbachia) of filarial nematodes.}, journal = {Parassitologia}, volume = {43}, number = {4}, pages = {173-178}, pmid = {12402526}, issn = {0048-2951}, mesh = {Animals ; Bacterial Proteins/genetics ; Brugia pahangi/*microbiology ; Computer Systems ; *Cytoskeletal Proteins ; DNA, Bacterial/*analysis ; Dirofilaria immitis/*microbiology ; Dirofilariasis/parasitology ; Dog Diseases/parasitology ; Dogs ; Female ; Gerbillinae/parasitology ; Male ; Polymerase Chain Reaction/*methods ; Reproducibility of Results ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/drug effects/genetics/*isolation & purification ; }, abstract = {Filarial nematodes harbour intracellular symbiotic bacteria belonging to the genus Wolbachia. Wolbachia is thought to play an important role in the biology of the nematode. Moreover, Wolbachia appears to be involved in the immunopathogenesis of filariasis and in the onset of the side-effects of antifilarial therapy. Investigations in these research areas require reliable methods to quantify Wolbachia both in nematodes and in vertebrate tissues. To this purpose, we designed a quantitative real-time PCR targeted on the ftsZ gene of the Wolbachia of Brugia pahangi, a model filarial species maintained in gerbils. The method was applied to quantify Wolbachia in Brugia pahangi, from animals with or without tetracycline treatment. Our results show that tetracycline treatment leads to dramatic reduction or clearance of Wolbachia from the nematode. Results obtained from different replicates were reproducible and the method appeared very sensitive compared to other PCR protocols for Wolbachia detection. Real-time PCR is thus an appropriate method for investigations on the biological role of Wolbachia and on the implication of these bacteria in the pathogenesis of filariasis. With slight modifications of the primers and probe, the protocol we have developed could be applied in studies of the human pathogen Brugia malayi and on the model filarial species Litomosoides sigmodontis.}, } @article {pmid12392911, year = {2002}, author = {Casiraghi, M and McCall, JW and Simoncini, L and Kramer, LH and Sacchi, L and Genchi, C and Werren, JH and Bandi, C}, title = {Tetracycline treatment and sex-ratio distortion: a role for Wolbachia in the moulting of filarial nematodes?.}, journal = {International journal for parasitology}, volume = {32}, number = {12}, pages = {1457-1468}, doi = {10.1016/s0020-7519(02)00158-3}, pmid = {12392911}, issn = {0020-7519}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Brugia pahangi/drug effects/*growth & development/*microbiology ; Female ; Gerbillinae/parasitology ; Immunohistochemistry ; Male ; Polymerase Chain Reaction ; *Sex Ratio ; Tetracycline/*pharmacology ; Time Factors ; Wolbachia/*drug effects/isolation & purification/*physiology ; }, abstract = {Filarial nematodes harbour intracellular bacteria of the genus Wolbachia. These bacteria are thought to be beneficial to the host nematode. Indeed, tetracycline treatments reduce the population of Wolbachia in filarial worms and have detrimental effects on the nematode. Even though various antibiotic-curing experiments have been performed on filariae, the actual role of Wolbachia in the biology of these nematodes is not yet clear. To address this issue, we designed a first experiment on a model filaria (Brugia pahangi), maintained in the gerbil (Meriones unguiculatus). In this experiment, timing of tetracycline treatment was set on the basis of the larval stage of the nematode. This first experiment showed that 2 weeks of treatment started after the L(4)-L(5) moult of males, but before the moult of females, led to significant sex-ratio distortion of the nematodes. We thus hypothesised that tetracycline interferes with the moult in B. pahangi. To test this hypothesis, we designed a second experiment in which antibiotic treatments were started (1). before the moult of both sexes, (2). after the moult of males but before the moult of females, or (3). after the moult of both sexes. Treatment 1 determined a reduction of worm recovery with no sex bias. Treatment 2 led to a male-biased sex-ratio. Treatment 3 had no effect on either worm recovery or sex-ratio. These results thus support the hypothesis that tetracycline treatment interferes with the L(4)-L(5) moult of B. pahangi. The nematodes recovered from the treated and control animals were examined for the presence of Wolbachia using both immunohistochemistry and real-time PCR. In general, nematodes from treated animals showed a dramatic reduction in Wolbachia content. In one group, Wolbachia depletion, as observed at the end of the treatment, was followed by a rebound to 'normal' values 160 days later. Prospects for antifilarial therapy using Wolbachia-targeted tetracycline treatments should thus take into account the possibility of Wolbachia rebound.}, } @article {pmid12389718, year = {2002}, author = {Charlat, S and Nirgianaki, A and Bourtzis, K and Merçot, H}, title = {Evolution of Wolbachia-induced cytoplasmic incompatibility in Drosophila simulans and D. sechellia.}, journal = {Evolution; international journal of organic evolution}, volume = {56}, number = {9}, pages = {1735-1742}, doi = {10.1111/j.0014-3820.2002.tb00187.x}, pmid = {12389718}, issn = {0014-3820}, mesh = {Animals ; *Biological Evolution ; Cytoplasm/*physiology ; DNA, Bacterial/genetics ; Drosophila/genetics/*microbiology/physiology ; Female ; Male ; Phylogeny ; Species Specificity ; Wolbachia/genetics/*physiology ; }, abstract = {The intracellular bacterium Wolbachia invades arthropod host populations through various mechanisms, the most common of which being cytoplasmic incompatibility (CI). CI involves elevated embryo mortality when infected males mate with uninfected females or females infected with different, incompatible Wolbachia strains. The present study focuses on this phenomenon in two Drosophila species: D. simulans and D. sechellia. Drosophila simulans populations are infected by several Wolbachia strains, including wHa and wNo. Drosophila sechellia is infected by only two Wolbachia: wSh and wSn. In both Drosophila species, double infections with Wolbachia are found. As indicated by several molecular markers, wHa is closely related to wSh, and wNo to wSn. Furthermore, the double infections in the two host species are associated with closely related mitochondrial haplotypes, namely siI (associated with wHa and wNo in D. simulans) and se (associated with wSh and wSn in D. sechellia). To test the theoretical prediction that Wolbachia compatibility types can diverge rapidly, we injected wSh and wSn into D. simulans, to compare their CI properties to those of their sister strains wHa and wNo, respectively, in the same host genetic background. We found that within each pair of sister strains CI levels were similar and that sister strains were fully compatible. We conclude that the short period for which the Wolbachia sister strains have been evolving separated from each other was not sufficient for their CI properties to diverge significantly.}, } @article {pmid12386340, year = {2002}, author = {Kondo, N and Nikoh, N and Ijichi, N and Shimada, M and Fukatsu, T}, title = {Genome fragment of Wolbachia endosymbiont transferred to X chromosome of host insect.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {99}, number = {22}, pages = {14280-14285}, pmid = {12386340}, issn = {0027-8424}, mesh = {Animals ; Base Sequence ; Blotting, Southern/methods ; Coleoptera/*genetics ; DNA, Bacterial ; Drosophila melanogaster/microbiology ; Drug Resistance, Bacterial ; Evolution, Molecular ; Female ; *Gene Transfer, Horizontal ; Genes, Bacterial ; *Genome, Bacterial ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/methods ; Rifampin/pharmacology ; Symbiosis ; Tetracycline/pharmacology ; Wolbachia/drug effects/*genetics/isolation & purification ; *X Chromosome ; }, abstract = {The adzuki bean beetle, Callosobruchus chinensis, is triple-infected with distinct lineages of Wolbachia endosymbiont, wBruCon, wBruOri, and wBruAus, which were identified by their wsp (Wolbachia surface protein) gene sequences. Whereas wBruCon and wBruOri caused cytoplasmic incompatibility of the host insect, wBruAus did not. Although wBruCon and wBruOri were easily eliminated by antibiotic treatments, wBruAus persisted over five treated generations and could not be eliminated. The inheritance pattern of wBruAus was, surprisingly, explained by sex-linked inheritance in male-heterozygotic organisms, which agreed with the karyotype of C. chinensis (2n = 20, XY). Quantitative PCR analysis demonstrated that females contain around twice as much wsp titer as males, which is concordant with an X chromosome linkage. Specific PCR and Southern blot analyses indicated that the wBruAus-bearing strain of C. chinensis contains only a fraction of the Wolbachia gene repertoire. Several genome fragments of wBruAus were isolated using an inverse PCR technique. The fragments exhibited a bacterial genome structure containing a number of ORFs typical of the alpha-proteobacteria, although some of the ORFs contained disruptive mutations. In the flanking region of ftsZ gene, a non-long terminal repeat (non-LTR) retrotransposon sequence, which is typical of insects but not found from bacteria, was present. These results strongly suggest that wBruAus has no microbial entity but is a genome fragment of Wolbachia endosymbiont transferred to the X chromosome of the host insect.}, } @article {pmid12296954, year = {2002}, author = {Tsuchida, T and Koga, R and Shibao, H and Matsumoto, T and Fukatsu, T}, title = {Diversity and geographic distribution of secondary endosymbiotic bacteria in natural populations of the pea aphid, Acyrthosiphon pisum.}, journal = {Molecular ecology}, volume = {11}, number = {10}, pages = {2123-2135}, doi = {10.1046/j.1365-294x.2002.01606.x}, pmid = {12296954}, issn = {0962-1083}, mesh = {Animals ; Aphids/*microbiology ; DNA, Ribosomal/analysis ; Female ; Genetic Variation/*genetics ; *Genetics, Population ; Japan ; Pisum sativum/*parasitology ; Polymerase Chain Reaction ; Proteobacteria/*classification/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Spiroplasma/*classification/genetics/isolation & purification ; Symbiosis ; }, abstract = {In addition to the essential intracellular symbiotic bacterium Buchnera, several facultative endosymbiotic bacteria called collectively secondary symbionts (S-symbionts) have been identified from the pea aphid Acyrthosiphon pisum. We conducted an extensive and systematic survey of S-symbionts in Japanese local populations of A. pisum using a specific PCR detection technique. Five S-symbionts of A. pisum, PASS, PAUS, PABS, Rickettsia and Spiroplasma, and two facultative endosymbionts universally found in various insects, Wolbachia and Arsenophonus, were targeted. Of 119 isofemale strains originating from 81 localities, 66.4% of the strains possessed either of four S-symbionts: PASS (38.7%); PAUS (16.0%); Rickettsia (8.4%); and Spiroplasma (3.4%), while 33.6% of the strains contained only Buchnera. PABS, Wolbachia and Arsenophonus were not detected from the Japanese strains of A. pisum. In order to understand intra- and interpopulational diversity of S-symbiont microbiota in detail, 858 insects collected from 43 localities were examined for infection with the four S-symbionts. It was demonstrated that different S-symbionts coexist commonly in the same local populations, but double infections with two S-symbionts were rarely detected. Notably, the S-symbionts exhibited characteristic geographical distribution patterns: PASS at high frequencies all over Japan; PAUS at high frequencies mainly in the northeastern part of Japan; and Rickettsia and Spiroplasma at low frequencies sporadically in the southwestern part of Japan. These results indicate that the geographical distribution and infection frequency of the S-symbionts, in particular PAUS, might be affected by environmental and/or historical factors. Statistical analyses suggested that the distribution of PAUS infection might be related to host plant species, temperature and precipitation.}, } @article {pmid12243742, year = {2002}, author = {Rao, RU and Moussa, H and Weil, GJ}, title = {Brugia malayi: effects of antibacterial agents on larval viability and development in vitro.}, journal = {Experimental parasitology}, volume = {101}, number = {1}, pages = {77-81}, doi = {10.1016/s0014-4894(02)00019-x}, pmid = {12243742}, issn = {0014-4894}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Azithromycin/pharmacology ; Brugia malayi/*drug effects/growth & development/microbiology/physiology ; Chloramphenicol/pharmacology ; Dose-Response Relationship, Drug ; Doxycycline/pharmacology ; Larva/drug effects/growth & development/physiology ; Molting/drug effects ; Movement/drug effects ; Rifampin/pharmacology ; Tetracycline/pharmacology ; Time Factors ; Wolbachia/drug effects/isolation & purification ; }, abstract = {Recent studies have suggested that intracellular Wolbachia bacteria are necessary for reproduction and survival of adult filarial worms. We now report results of in vitro studies of effects of antibacterial antibiotics (tetracycline, rifampicin, chloramphenicol, azithromycin, and doxycycline) on Brugia malayi infective larvae (L3) motility and molting. All of the antibiotics tested except chloramphenicol decreased L3 motility by 50% or more at 10 days, with minimal effective concentrations (MECs) of 20-100 microg/ml. Tetracyclines, rifampicin, and chloramphenicol inhibited L3 to L4 molting by 12 days in a concentration- and time-dependent manner, with MECs in the range of 1-20 microg/ml. These studies show that antibiotics active against Rickettsiaceae inhibit B. malayi L3 molting at low concentrations in vitro; higher concentrations kill the larvae. While it is possible that antibiotics directly affect filarial L3, we believe it is more likely that the effects seen are indirect effects related to bacterial killing.}, } @article {pmid12234541, year = {2002}, author = {Stouthamer, R and Mak, F}, title = {Influence of antibiotics on the offspring production of the Wolbachia-infected parthenogenetic parasitoid Encarsia formosa.}, journal = {Journal of invertebrate pathology}, volume = {80}, number = {1}, pages = {41-45}, doi = {10.1016/s0022-2011(02)00034-4}, pmid = {12234541}, issn = {0022-2011}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Female ; Fertility ; Longevity ; Male ; Sex Ratio ; Tetracycline/*pharmacology ; Wasps/*drug effects/growth & development/microbiology ; Wolbachia/*physiology ; }, abstract = {Three different concentrations of the antibiotic tetracycline in honey were tested for their influence on the offspring production and longevity of the parasitoid wasp Encarsia formosa. Several earlier publications did not provide a conclusive answer on the effect that the Wolbachia have on these wasps. The results of our experiments show that at high tetracycline hydrochloride concentrations in honey (50mg/ml) the antibiotic is toxic to the females, all females died within three days after the antibiotic treatment. The concentration 5mg/ml was less toxic although the treated females also lived shorter and produced less offspring than the control females. At the lowest tested concentration of 1mg/ml there was no significant difference either in offspring production or in longevity between the control and the treated females. The antibiotic treatment at both 5 and 1mg/ml resulted in exclusively male progeny after the first two days of oviposition. These results are consistent with the theory that in species in which all individuals are infected the Wolbachia should not impose a large fitness cost.}, } @article {pmid12234537, year = {2002}, author = {Grenier, S and Gomes, SM and Pintureau, B and Lassablière, F and Bolland, P}, title = {Use of tetracycline in larval diet to study the effect of Wolbachia on host fecundity and clarify taxonomic status of Trichogramma species in cured bisexual lines.}, journal = {Journal of invertebrate pathology}, volume = {80}, number = {1}, pages = {13-21}, doi = {10.1016/s0022-2011(02)00039-3}, pmid = {12234537}, issn = {0022-2011}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Crosses, Genetic ; Female ; Fertility/drug effects ; Larva/drug effects/microbiology ; Male ; Tetracycline/*pharmacology ; Wasps/drug effects/*microbiology ; Wolbachia/*physiology ; }, abstract = {Endosymbionts of the genus Wolbachia were efficiently cured from Trichogramma species by incorporating 0.02% tetracycline into the artificial diet used to rear larvae. Use of this technique yielded stable cured lines (bisexual and arrhenotokous lines) in which no Wolbachia organisms were detected by PCR for up to 14 generations after curing. Four cured strains of Trichogramma pretiosum showed a significantly lower total fecundity compared to their Wolbachia-infected counterpart. However, the fecundity of a single cured strain of Trichogramma evanescens was similar to its Wolbachia-infected counterpart. These differences in the effect on fecundity may be due to differences between the Wolbachia strains infecting T. pretiosum or T. evanescens, providing additional evidence for the hypothesis that a specific interaction exists between some Trichogramma species and their Wolbachia symbionts. Tetracycline in larval diet was also used to generate bisexual strains of Trichogramma oleae and Trichogramma cordubensis so that these species could be crossed with the closely related species, respectively, T. pretiosum and T. evanescens, to test their compatibility. These crosses showed a lack of compatibility, validating maintenance of these as distinct species.}, } @article {pmid12234534, year = {2002}, author = {Taylor, MJ}, title = {Wolbachia endosymbiotic bacteria of filarial nematodes. A new insight into disease pathogenesis and control.}, journal = {Archives of medical research}, volume = {33}, number = {4}, pages = {422-424}, doi = {10.1016/s0188-4409(02)00377-6}, pmid = {12234534}, issn = {0188-4409}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Filariasis/drug therapy/*immunology/microbiology/parasitology ; Filarioidea/cytology/*microbiology ; Humans ; Inflammation ; Symbiosis ; Wolbachia/*immunology/physiology ; }, abstract = {Filarial nematodes are parasitic worms that cause some of the most devastating of all tropical diseases such as elephantiasis and river blindness. Studies on the inflammatory pathogenesis of filarial disease have shown that endotoxin-like activity derived from endosymbiotic Wolbachia bacteria is the major inflammatory stimulus of filarial nematodes. Wolbachia appear to have evolved as essential symbionts of their filarial nematode hosts. Antibiotic depletion of bacteria shows that they are required for normal fertility and development of the worm and may even protect the parasites from host immunity. In addition to the uncovering of a fascinating symbiotic relationship, this discovery means we can now consider using antibiotics as a new approach to the treatment of filarial diseases.}, } @article {pmid12221039, year = {2003}, author = {Normark, BB}, title = {The evolution of alternative genetic systems in insects.}, journal = {Annual review of entomology}, volume = {48}, number = {}, pages = {397-423}, doi = {10.1146/annurev.ento.48.091801.112703}, pmid = {12221039}, issn = {0066-4170}, mesh = {Animals ; *Biological Evolution ; Diploidy ; Disorders of Sex Development ; Extrachromosomal Inheritance ; Female ; Haploidy ; Insecta/*genetics/microbiology ; Male ; Models, Genetic ; Parthenogenesis ; Sex Determination Processes ; Wolbachia/genetics ; }, abstract = {There are three major classes of insect genetic systems: those with diploid males (diplodiploidy), those with effectively haploid males (haplodiploidy), and those without males (thelytoky). Mixed systems, involving cyclic or facultative switching between thelytoky and either of the other systems, also occur. I present a classification of the genetic systems of insects and estimate the number of evolutionary transitions between them that have occurred. Obligate thelytoky has arisen from each of the other systems, and there is evidence that over 900 such origins have occurred. The number of origins of facultative thelytoky and the number of reversions from obligate thelytoky to facultative and cyclic thelytoky are difficult to estimate. The other transitions are few in number: five origins of cyclic thelytoky, eight origins of obligate haplodiploidy (including paternal genome elimination), the strange case of Micromalthus, and the two reversions from haplodiploidy to diplodiploidy in scale insects. Available evidence tends to support W.D. Hamilton's hypothesis that maternally transmitted endosymbionts have been involved in the origins of haplodiploidy. Bizarre systems of extrazygotic inheritance in Sternorrhyncha are not easily accommodated into any existing classification of genetic systems.}, } @article {pmid12219740, year = {2002}, author = {Callahan, GN}, title = {Madness.}, journal = {Emerging infectious diseases}, volume = {8}, number = {9}, pages = {998-1002}, doi = {10.3201/eid0809.ad0809}, pmid = {12219740}, issn = {1080-6040}, mesh = {Animals ; Ants/parasitology/physiology ; Borna disease virus/physiology ; Cats ; Depression/etiology/physiopathology ; Dicrocoelium/physiology ; Female ; Humans ; Male ; Mental Disorders/*etiology/*microbiology/physiopathology/virology ; Neurosyphilis/complications ; Paintings ; Phthiraptera/parasitology/physiology ; Rats ; Toxoplasma/physiology ; Wolbachia/physiology ; }, } @article {pmid12206235, year = {2002}, author = {Vala, F and Weeks, A and Claessen, D and Breeuwer, JA and Sabelis, MW}, title = {Within- and between-population variation for Wolbachia-induced reproductive incompatibility in a haplodiploid mite.}, journal = {Evolution; international journal of organic evolution}, volume = {56}, number = {7}, pages = {1331-1339}, doi = {10.1111/j.0014-3820.2002.tb01447.x}, pmid = {12206235}, issn = {0014-3820}, mesh = {Animals ; Cytoplasm/*microbiology/physiology ; Genetic Variation ; Male ; Mites/*microbiology/physiology ; Models, Theoretical ; Reproduction/physiology ; Wolbachia/genetics/pathogenicity/*physiology ; }, abstract = {Wolbachia pipientis is a bacterium that induces cytoplasmic incompatibility (CI), the phenomenon in which infected males are reproductively incompatible with uninfected females. CI spreads in a population of hosts because it reduces the fitness of uninfected females relative to infected females. CI encompasses two steps: modification (mod) of sperm of infected males and rescuing (resc) of these chromosomes by Wolbachia in the egg. Infections associated with CI have mod+ resa+ phenotypes. However, mod- resc+ phenotypes also exist; these do not result in CI. Assuming mod/resc phenotypes are properties of the symbiont, theory predicts that mod- resc+ infections can only spread in a host population where a mod+ resc+ infection already occurs. A mod- resc+ infection spreads if the cost it imposes on the infected females is lower than the cost inflicted by the resident (mod+ resc+) infection. Furthermore, introduction of a mod- Wolbachia eventually drives infection to extinction. The uninfected population that results can be recolonized by a CI-causing Wolbachia. Here, we investigated whether variability for induction of CI was present in two Tetranychus urticae populations. In one population all isofemale lines tested were mod-. In the other, mod+ resc+ and mod- resc+ isofemale lines coexisted. We found no evidence for a cost difference to females expressing either type (mod-/-). Infections in the two populations could not be distinguished based on sequences of two Wolbachia genes. We consider the possibility that mod- is a host effect through a population dynamics model. A mod- host allele leads to infection extinction in the absence of fecundity differences. Furthermore, the uninfected population that results is immune to reestablishment of the (same) CI-causing Wolbachia.}, } @article {pmid12208043, year = {2002}, author = {Egyed, Z and Sréter, T and Széll, Z and Nyiro, G and Márialigeti, K and Varga, I}, title = {Molecular phylogenetic analysis of Onchocerca lupi and its Wolbachia endosymbiont.}, journal = {Veterinary parasitology}, volume = {108}, number = {2}, pages = {153-161}, doi = {10.1016/s0304-4017(02)00186-3}, pmid = {12208043}, issn = {0304-4017}, mesh = {Animals ; Bacterial Outer Membrane Proteins/chemistry/genetics ; Bacterial Proteins/chemistry/genetics ; Base Sequence ; *Cytoskeletal Proteins ; Dog Diseases/parasitology ; Dogs ; Electron Transport Complex IV/chemistry/genetics ; Molecular Sequence Data ; Onchocerca/*genetics/growth & development ; Onchocerciasis, Ocular/parasitology/veterinary ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Symbiosis ; Wolbachia/*genetics ; }, abstract = {The morphology of Onchocerca lupi, responsible for canine ocular onchocercosis, is unique within the genus. Earlier analyses of the 5S ribosomal RNA gene spacer region sequence of the parasite and the 16S ribosomal RNA gene sequence of its Wolbachia endosymbiotic bacteria (Rickettsiales) supported the morphological and biological arguments that O. lupi is a distinct species. However, the exact phylogenetic position of O. lupi and its endosymbiont could not be unambiguously determined. Herein we report analyses based on the mitochondrial cytochrome oxidase I (COI) gene of the filarial species and the Wolbachia surface protein (wsp) and the bacterial cell-cycle ftsZ genes of their wolbachiae. Our results indicate that O. lupi separated from other Onchocerca spp. early in evolution. This is in line with the previous morphological analysis demonstrating that O. lupi is an atypical Onchocerca species showing both primitive and evolved characters. The phylogenetic trees generated for the COI sequences of filariae and the wsp and ftsZ sequences of their wolbachiae were congruent with each other, which supports the hypothesis that nematodes and their Wolbachia endobacteria share a long co-evolutionary history.}, } @article {pmid12207731, year = {2002}, author = {Rokas, A and Atkinson, RJ and Nieves-Aldrey, JL and West, SA and Stone, GN}, title = {The incidence and diversity of Wolbachia in gallwasps (Hymenoptera; Cynipidae) on oak.}, journal = {Molecular ecology}, volume = {11}, number = {9}, pages = {1815-1829}, doi = {10.1046/j.1365-294x.2002.01556.x}, pmid = {12207731}, issn = {0962-1083}, mesh = {Animals ; Female ; Genes, Bacterial ; Male ; Phylogeny ; Quercus/*parasitology ; Wasps/classification/*microbiology/physiology ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia bacteria infect approximately 20% of all insect species, and cause a range of alterations to host reproduction, including imposition of thelytoky. The incidence and phenotypic impact of Wolbachia remains to be established in many insect taxa, and considerable research effort is currently focused on its association with particular reproductive modes and the relative importance of the various pathways via which infection occurs. Gallwasps represent an attractive system for addressing these issues for two reasons. First, they show a diversity of reproductive modes (including arrhenotoky, thelytoky and cyclical parthenogenesis) in which the impact of Wolbachia infection can be examined. Second, they occupy two intimately linked trophic niches (gall-inducers and inquilines) between which there is potential for the horizontal exchange of Wolbachia infection. In the arrhenotokous gallwasp lineages screened to date (the herb-galling 'Aylacini' and the rose-galling Diplolepidini), Wolbachia infection always induces thelytoky. The impact of Wolbachia in other arrhenotokous clades, and in the cyclically parthenogenetic clades remains unknown. Here we use polymerase chain reaction (PCR) screening and sequence data for two Wolbachia genes (wsp and ftsZ) to examine the prevalence and incidence of Wolbachia infection in 64 species (a total of 609 individuals) in two further tribes: the arrhenotokous inquilines (tribe Synergini), and the cyclically parthenogenetic oak gallwasps (tribe Cynipini). We ask: (i) whether Wolbachia infection has any apparent impact on host reproduction in the two tribes and (ii) whether there is any correlation between Wolbachia infection and the apparent lack of an arrhenotokous generation in many oak gallwasp life cycles. We show: (i) that Wolbachia infection is rare in the Cynipini. Infected species show no deviation from cyclical parthenogenesis, and infection is no more common in species known only from a thelytokous generation; (ii) that there is a higher incidence of infection within the arrhenotokous inquilines, and generally in gallwasp tribes without cyclical parthenogensis; (iii) all Wolbachia-positive inquiline species are known to possess males, implying either that Wolbachia infection does not result in loss of sex in this tribe or, more probably, that (as for some rose gallwasps) Wolbachia infection leads to loss of sex in specific populations; and (iv) although we find some inquilines and gall inducers to be infected with Wolbachia having the same wsp sequence, these hosts are not members of the same gall communities, arguing against frequent horizontal transmission between these two trophic groups. We suggest that exchange may be mediated by the generalist parasitoids common in oak galls.}, } @article {pmid12200493, year = {2002}, author = {Jiggins, FM}, title = {The rate of recombination in Wolbachia bacteria.}, journal = {Molecular biology and evolution}, volume = {19}, number = {9}, pages = {1640-1643}, doi = {10.1093/oxfordjournals.molbev.a004228}, pmid = {12200493}, issn = {0737-4038}, mesh = {Alleles ; Animals ; Arthropods/microbiology ; Gene Frequency/genetics ; Likelihood Functions ; Recombination, Genetic/*genetics ; Wolbachia/*genetics ; }, } @article {pmid12192522, year = {2002}, author = {Jamnongluk, W and Kittayapong, P and Baimai, V and O'Neill, SL}, title = {Wolbachia infections of tephritid fruit flies: molecular evidence for five distinct strains in a single host species.}, journal = {Current microbiology}, volume = {45}, number = {4}, pages = {255-260}, doi = {10.1007/s00284-002-3746-1}, pmid = {12192522}, issn = {0343-8651}, mesh = {Amino Acid Sequence ; Animals ; Cloning, Molecular ; DNA Primers ; Genes, Bacterial ; Host-Parasite Interactions ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods ; Sequence Alignment ; Tephritidae/*microbiology ; Wolbachia/*classification/genetics/growth & development/isolation & purification ; }, abstract = {Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and can induce cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing or feminization in their hosts. Here, we report phylogenetic relationships of Wolbachia in tephritid fruit flies based on wsp gene sequences. We also report, for the first time, five distinct strains of Wolbachia in Bactrocera ascita sp. B. Four of the five Wolbachia strains found in this species were in the same groups as those found in other tephritid fruit flies, suggesting possible horizontal transmission of Wolbachia from other fruit flies into B. ascita sp. B. The unreliability of wsp-specific group primers demonstrated in this study suggests that these primers might be useful only for preliminary identification of Wolbachia. Final determination of group affiliation needs to be verified with wsp sequence data.}, } @article {pmid12180086, year = {2002}, author = {Kageyama, D and Nishimura, G and Hoshizaki, S and Ishikawa, Y}, title = {Feminizing Wolbachia in an insect, Ostrinia furnacalis (Lepidoptera: Crambidae).}, journal = {Heredity}, volume = {88}, number = {6}, pages = {444-449}, doi = {10.1038/sj.hdy.6800077}, pmid = {12180086}, issn = {0018-067X}, mesh = {Animals ; DNA, Bacterial/genetics ; Female ; Feminization/microbiology ; Lepidoptera/*microbiology ; Male ; Phylogeny ; Sex Ratio ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Wolbachia, which forms a group of maternally inherited bacteria in arthropods, often cause reproduction alterations in their hosts, such as cytoplasmic incompatibility, parthenogenesis, male-killing, hybrid breakdown and feminization. To date, Wolbachia-induced feminization has been reported only in isopods. Here we report that a Wolbachia strain feminizes an insect host, Ostrinia furnacalis. Among 79 wild females of O. furnacalis examined, Wolbachia infection was detected in 13 females. Twelve of the 13 infected females produced all-female progenies, and this trait was maternally inherited. Tetracycline treatment of thelygenic matrilines resulted in the production of all-male progenies. The present findings indicate that the Wolbachia infection induces feminization of genetic males in O. furnacalis. Differences in the Wolbachia-induced feminization in O. furnacalis and that in isopods are discussed along with the differences in sex determination mechanisms between insects and isopods. Phylogenetic analysis of the wsp gene sequence of Wolbachia suggests independent evolutionary origins for the Wolbachia-induced feminizations in O. furnacalis and in isopods. Our findings over 5 years suggest that the infection has been maintained at a low prevalence in the O. furnacalis population.}, } @article {pmid12147509, year = {2002}, author = {Ijichi, N and Kondo, N and Matsumoto, R and Shimada, M and Ishikawa, H and Fukatsu, T}, title = {Internal spatiotemporal population dynamics of infection with three Wolbachia strains in the adzuki bean beetle, Callosobruchus chinensis (Coleoptera: Bruchidae).}, journal = {Applied and environmental microbiology}, volume = {68}, number = {8}, pages = {4074-4080}, pmid = {12147509}, issn = {0099-2240}, mesh = {Animals ; Coleoptera/growth & development/*microbiology/ultrastructure ; DNA, Bacterial/analysis ; Fabaceae/parasitology ; Female ; In Situ Hybridization ; Male ; Microscopy, Electron ; Polymerase Chain Reaction ; Population Dynamics ; Wolbachia/classification/genetics/*growth & development/*pathogenicity ; }, abstract = {The adzuki bean beetle, Callosobruchus chinensis, is infected with three distinct lineages of endosymbiotic bacteria belonging to the genus Wolbachia, which were designated wBruCon, wBruOri, and wBruAus. In an attempt to understand the mechanisms underlying the infection with these three organisms, the spatiotemporal infection dynamics of the three Wolbachia strains was investigated in detail by using a quantitative PCR technique. During the development of C. chinensis, the wBruCon, wBruOri, and wBruAus infection levels consistently increased but the growth patterns were different. The levels of infection plateaued at the pupal stage at approximately 3 x 10(8), 2 x 10(8), and 5 x 10(7) wsp copy equivalents per insect for wBruCon, wBruOri, and wBruAus, respectively. At the whole-insect level, the population densities of the three Wolbachia types did not show remarkable differences between adult males and females. At the tissue level, however, the total densities and relative levels of the three Wolbachia types varied significantly when different tissues and organs were compared and when the same tissues derived from males and females were compared. The histological data obtained by in situ hybridization and electron microscopy were concordant with the results of quantitative PCR analyses. Based on the histological data and the peculiar Wolbachia composition commonly found in nurse tissues and oocytes, we suggest that the Wolbachia strains are vertically transmitted to oocytes not directly, but by way of nurse tissue. On the basis of our results, we discuss interactions among the three coinfecting Wolbachia types, reproductive strategies of Wolbachia, and factors involved in the different cytoplasmic incompatibility phenotypes.}, } @article {pmid12144650, year = {2002}, author = {Jiggins, FM and Bentley, JK and Majerus, ME and Hurst, GD}, title = {Recent changes in phenotype and patterns of host specialization in Wolbachia bacteria.}, journal = {Molecular ecology}, volume = {11}, number = {8}, pages = {1275-1283}, doi = {10.1046/j.1365-294x.2002.01532.x}, pmid = {12144650}, issn = {0962-1083}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Butterflies/*microbiology ; Chaperonins ; Escherichia coli Proteins ; Heat-Shock Proteins/genetics ; Male ; Molecular Sequence Data ; Phenotype ; Phylogeny ; Sex Factors ; Symbiosis ; Wolbachia/classification/*genetics/pathogenicity/*physiology ; }, abstract = {Wolbachia are a genus of bacterial symbionts that are known to manipulate the reproduction of their arthropod hosts, both by distorting the host sex ratio and by inducing cytoplasmic incompatibility. Previous work has suggested that some Wolbachia clades specialize in particular host taxa, but others are diverse. Furthermore, the frequency with which related strains change in phenotype is unknown. We have examined these issues for Wolbachia bacteria from Acraea butterflies, where different interactions are known in different host species. We found that bacteria from Acraea butterflies mostly cluster together in several different clades on the bacterial phylogeny, implying specialization of particular strains on these host taxa. We also observed that bacterial strains with different phenotypic effects on their hosts commonly shared identical gene sequences at two different loci. This suggests both that the phenotypes of the strains have changed recently between sex ratio distortion and cytoplasmic incompatibility, and that host specialization is not related to the bacterial phenotype, as suggested from previous data. We also analysed published data from other arthropod taxa, and found that the Wolbachia infections of the majority of arthropod genera tend to cluster together on the bacterial phylogeny. Therefore, we conclude that Wolbachia is most likely to move horizontally between closely related hosts, perhaps because of a combination of shared vectors for transmission and physiological specialization of the bacteria on those hosts.}, } @article {pmid12144285, year = {2002}, author = {Ricci, I and Cancrini, G and Gabrielli, S and D'Amelio, S and Favi, G}, title = {Searching for Wolbachia (Rickettsiales: Rickettsiaceae) in mosquitoes (Diptera: Culicidae): large polymerase chain reaction survey and new identifications.}, journal = {Journal of medical entomology}, volume = {39}, number = {4}, pages = {562-567}, doi = {10.1603/0022-2585-39.4.562}, pmid = {12144285}, issn = {0022-2585}, mesh = {Aedes/microbiology ; Animals ; Anopheles/microbiology ; Base Sequence ; Culex/microbiology ; Culicidae/*microbiology ; DNA, Bacterial ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods ; Wolbachia/classification/*genetics ; }, abstract = {Bacteria of the genus Wolbachia constitute a group of intracellular and maternally inherited micro-organisms that are widespread in arthropods, inducing several reproductive disorders such as cytoplasmic incompatibility in their hosts. Considering relevant biological implications related to the presence of Wolbachia in several insect orders, for example its potential role as mechanism for rapid speciation and as vehicle to drive genetic markers in wild populations of vectors of medical and veterinary interest, we carried out an extensive polymerase chain reaction survey to detect Wolbachia in several species of mosquito belonging to genera involved in the transmission of pathogens. Five species out of 26 tested have shown to be infected; for four of them this is the first evidence of the Wolbachia infection. A phylogenetic analysis was also performed, positioning the five Wolbachia strains in the phyletic subdivision B.}, } @article {pmid12144012, year = {2002}, author = {Egas, M and Vala, F and Breeuwer, JA}, title = {On the evolution of cytoplasmic incompatibility in haplodiploid species.}, journal = {Evolution; international journal of organic evolution}, volume = {56}, number = {6}, pages = {1101-1109}, doi = {10.1111/j.0014-3820.2002.tb01424.x}, pmid = {12144012}, issn = {0014-3820}, mesh = {Bacterial Infections/genetics ; *Biological Evolution ; Cytoplasm/*physiology ; Female ; *Haploidy ; Humans ; Male ; Species Specificity ; Wolbachia/*genetics/growth & development/pathogenicity ; }, abstract = {The most enigmatic sexual manipulation by Wolbachia endosymbionts is cytoplasmic incompatibility (CI): infected males are reproductively incompatible with uninfected females. In this paper, we extend the theory on population dynamics and evolution of CI, with emphasis on haplodiploid species. First, we focus on the problem of the threshold to invasion of the Wolbachia infection in a population. Simulations of the dynamics of infection in small populations show that it does not suffice to assume invasion by drift alone (or demographic "accident"). We propose several promising alternatives that may facilitate invasion of Wolbachia in uninfected populations: sex-ratio effects, meta population structure, and other fitness-compensating effects. Including sex-ratio effects of Wolbachia allows invasion whenever infected females produce more infected daughters than uninfected females produce uninfected daughters. Several studies on haplodiploid species suggest the presence of such sex-ratio effects. The simple metapopulation model we analyzed predicts that, given that infecteds are better "invaders," uninfecteds must be better "colonizers" to maintain coexistence of infected and uninfected patches. This condition seems more feasible for species that suffer local extinction due to predation (or parasitization) than for species that suffer local extinction due to overexploiting their resource(s). Finally, we analyze the evolution of CI in haplodiploids once a population has been infected. Evolution does not depend on the type of CI (female mortality or male production), but hinges solely on decreasing the fitness cost and/or increasing the transmission efficiency. Our models offer new perspectives for increasing our understanding of the population and evolutionary dynamics of CI.}, } @article {pmid12140246, year = {2002}, author = {Jiggins, FM and Hurst, GD and Yang, Z}, title = {Host-symbiont conflicts: positive selection on an outer membrane protein of parasitic but not mutualistic Rickettsiaceae.}, journal = {Molecular biology and evolution}, volume = {19}, number = {8}, pages = {1341-1349}, doi = {10.1093/oxfordjournals.molbev.a004195}, pmid = {12140246}, issn = {0737-4038}, mesh = {Amino Acid Sequence ; Animals ; *Antigens, Bacterial ; Bacterial Outer Membrane Proteins/chemistry/*genetics ; Ehrlichia ruminantium/classification/*genetics/physiology ; Likelihood Functions ; Molecular Sequence Data ; Phylogeny ; *Selection, Genetic ; *Symbiosis ; Wolbachia/classification/*genetics/physiology ; }, abstract = {The Rickettsiaceae is a family of intracellular bacterial symbionts that includes both vertically transmitted parasites that spread by manipulating the reproduction of their host (Wolbachia in arthropods) and horizontally transmitted parasites (represented by Cowdria ruminantium), and mutualists (Wolbachia pipientis in nematode worms). We have investigated the nature of natural selection acting on an outer membrane protein, the wsp gene in Wolbachia and its homologue map1 in Cowdria, thought likely to be involved in host-parasite interactions in these bacteria. The ratio of nonsynonymous to synonymous substitution rates (d(N)/d(S)) at individual amino acid sites or at lineages within the gene's phylogeny was estimated using maximum likelihood models of codon substitution. The first hypothesis we tested was that this protein is under positive selection in the parasitic but not in the mutualistic Rickettsiaceae. This hypothesis was supported as positive selection and was detected in Cowdria and arthropod Wolbachia sequence evolution but not in the evolution of Wolbachia sequences from nematodes. Furthermore, this selection was concentrated outside the transmembrane region of the protein and, therefore, in the regions of the protein that may interact with the host. The second hypothesis tested was that positive selection would be stronger in the strains of arthropod Wolbachia that distort the host sex ratio than in those that induce cytoplasmic incompatibility. However, we found no support for this hypothesis. In conclusion, our results are consistent with the hypothesis that antagonistic coevolution causes faster evolution of surface protein sequences in parasites than in mutualists. Confirmation of this conclusion awaits the replication of these results both in additional genes and across more bacterial taxa. The regions of the wsp and map1 genes we identified as likely to be involved in host-parasite arms races should be examined in future studies of parasite virulence and host immune responses, and during the design of vaccines.}, } @article {pmid12136416, year = {2002}, author = {Oliver, P and Castro, JA and Picornell, A and Ramon, MM and Solé, E and Balanyà, J and Serra, L and Latorre, A and Moya, A}, title = {Linkage disequilibria between mtDNA haplotypes and chromosomal arrangements in a natural population of Drosophila subobscura.}, journal = {Heredity}, volume = {89}, number = {2}, pages = {133-138}, doi = {10.1038/sj.hdy.6800116}, pmid = {12136416}, issn = {0018-067X}, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics ; Genetics, Population ; Haplotypes ; *Linkage Disequilibrium ; }, abstract = {The association between mtDNA haplotypes and chromosomal arrangements in a natural population of Drosophila subobscura from Calvia (Balearic Islands, Spain) was studied in order to search for linkage disequilibria, in an attempt to explain the populational dynamics of the mtDNA haplotypes of this species in nature. The presence of Wolbachia was not detected. Two main haplotypes (I and II) were found, as well as a series of less common ones. The Tajima D-test seemed to indicate some kind of seasonal population bottleneck. An analysis of linkage disequilibrium and factorial analysis of correspondences detected an association between haplotype I and the J(ST) inversion and haplotype II and the J(1) inversion.}, } @article {pmid12135259, year = {2002}, author = {Kitrayapong, P and Baimai, V and O'Neill, SL}, title = {Field prevalence of Wolbachia in the mosquito vector Aedes albopictus.}, journal = {The American journal of tropical medicine and hygiene}, volume = {66}, number = {1}, pages = {108-111}, doi = {10.4269/ajtmh.2002.66.108}, pmid = {12135259}, issn = {0002-9637}, mesh = {Aedes/*microbiology ; Animals ; Bacterial Outer Membrane Proteins/chemistry/genetics ; DNA, Bacterial/chemistry/genetics ; Female ; Insect Vectors/*microbiology ; Polymerase Chain Reaction ; Prevalence ; Thailand ; Wolbachia/classification/genetics/*growth & development ; }, abstract = {The endosymbiotic bacteria in the genus Wolbachia have been proposed as a potential candidate to deliver pathogen-blocking genes into natural populations of medically important insects. The successful application of Wolbachia in insect vector control depends on the ability of the agent to successfully invade and maintain itself at high frequency under field conditions. Here, we evaluated the prevalence of Wolbachia infections in a field population of the Wolbachia-superinfected mosquito Aedes albopictus. A field prevalence of 100% (n = 1,016) was found in a single population in eastern Thailand via polymerase chain reaction (PCR) testing of Wolbachia both from individual parent females and their corresponding F1 offspring. This is the first report of accurate Wolbachia prevalence in a field population of an insect disease vector. The prevalence of superinfection was estimated to be 99.41%. All single-infected individual mosquitoes (n = 6) were found to harbor group A Wolbachia. For this particular population, none was found to be single-infected with group B Wolbachia. Our results also show that PCR testing of field materials alone without checking F1 offspring overestimated the natural prevalence of single infection. Thus, the confirmation of infection status by means of F1 offspring was critical to the accurate estimates of Wolbachia prevalence under field conditions.}, } @article {pmid12135258, year = {2002}, author = {Kittayapong, P and Baisley, KJ and Sharpe, RG and Baimai, V and O'Neill, SL}, title = {Maternal transmission efficiency of Wolbachia superinfections in Aedes albopictus populations in Thailand.}, journal = {The American journal of tropical medicine and hygiene}, volume = {66}, number = {1}, pages = {103-107}, doi = {10.4269/ajtmh.2002.66.103}, pmid = {12135258}, issn = {0002-9637}, support = {G0700837/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; DNA, Bacterial/chemistry/genetics ; Female ; Insect Vectors/*microbiology ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Thailand ; Wolbachia/genetics/*growth & development ; }, abstract = {We examined the transmission efficiency of 2 strains of Wolbachia bacteria that cause cytoplasmic incompatibility in field populations of Aedes albopictus by polymerase chain reaction assay. We found mainland and island populations throughout Thailand to be superinfected with group A and B bacteria. Of 320 Wolbachia-positive adult mosquitoes, 97.5% were infected with both groups. Single infected individuals of each Wolbachia group were encountered in nearly equal numbers. We screened 550 offspring from 80 field-collected mothers and found the transmission efficiency of group A Wolbachia to be 96.7% and that of group B Wolbachia to be 99.6%. Mothers that did not transmit both Wolbachia infections to all of their offspring were significantly larger in size than those with perfect transmission fidelity. We discuss our findings in relation to the prospects of the use of Wolbachia as a gene-driving mechanism.}, } @article {pmid12132134, year = {2002}, author = {Kager, PA}, title = {[Combatting river blindness by means of chemotherapy directed at the symbiotic Wolbachia bacteria in the causative filariae].}, journal = {Nederlands tijdschrift voor geneeskunde}, volume = {146}, number = {26}, pages = {1212-1215}, pmid = {12132134}, issn = {0028-2162}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology/*therapeutic use ; Disease Models, Animal ; Doxycycline/pharmacology/*therapeutic use ; *Drosophila Proteins ; Female ; Humans ; Male ; Membrane Glycoproteins/physiology ; Mice ; Onchocerca/microbiology/*physiology ; Onchocerciasis, Ocular/*drug therapy/prevention & control ; Receptors, Cell Surface/physiology ; Symbiosis ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Wolbachia/*drug effects/physiology ; }, abstract = {In a mouse model of river blindness it was demonstrated that Wolbachia bacteria, endosymbionts of filarial nematodes, play an important role in the inflammatory process leading to the disease and that this process depends on Toll-like receptor 4. Wolbachia is found in many arthropods and in all filariae pathogenic for man. Treatment with doxycycline depletes female filariae of Wolbachia and renders them infertile for at least 18 months. Chemotherapy of filarial nematodes should be studied for the reduction and possibly prevention of pathology due to filariae, and for a contribution to control and eradication programmes.}, } @article {pmid12102035, year = {2002}, author = {Hassler, D and Kimmig, P and Braun, R}, title = {[River blindness: not only the work of Onchocerca volvulus?].}, journal = {Deutsche medizinische Wochenschrift (1946)}, volume = {127}, number = {22}, pages = {1178-1179}, pmid = {12102035}, issn = {0012-0472}, mesh = {Animals ; Female ; Humans ; Inflammation/etiology ; Male ; Mammals ; Onchocerca volvulus/*physiology ; Onchocerciasis, Ocular/drug therapy/*etiology/pathology ; *Symbiosis ; Wolbachia/*physiology ; }, } @article {pmid12099435, year = {2002}, author = {Rao, R and Well, GJ}, title = {In vitro effects of antibiotics on Brugia malayi worm survival and reproduction.}, journal = {The Journal of parasitology}, volume = {88}, number = {3}, pages = {605-611}, doi = {10.1645/0022-3395(2002)088[0605:IVEOAO]2.0.CO;2}, pmid = {12099435}, issn = {0022-3395}, support = {AI 02642/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology/therapeutic use ; Brugia malayi/*drug effects/physiology ; Female ; Filariasis/drug therapy ; Formazans/chemistry ; Gerbillinae ; Histocytochemistry ; Male ; Microfilariae ; Microscopy, Electron ; Reproduction/drug effects ; Tetrazolium Salts/chemistry ; }, abstract = {Recent studies have suggested that intracellular Wolbachia spp. endobacteria are necessary for the reproduction and survival of filarial nematodes. The effects of antibiotics that are active against related bacteria on adult worms and microfilariae (Mf) of Brugia malayi in vitro were investigated. Antibiotics tested were doxycycline (Doxy), tetracycline (Tet), rifampicin (Rif), azithromycin (Azith), and chloramphenicol (Chlor). Doxy, Tet, Rif, and Azith reduced release of Mf by adult female worms. The minimum effective concentrations that reduced Mf release by 50% were 5 microg/ml for Doxy, 20 microg/ml for Tet, 40 microg/ml for Rif, and 100 microg/ml for Azith. The same drugs (at higher concentrations) killed adult worms and Mf. Embryograms showed that Tets blocked embryogenesis in female worms. Electron microscopy (EM) showed that the Tets, Rif, and Azith cleared Wolbachia spp. from adult worms and damaged developing embryos. These studies show that antibiotics active against Rickettsiaceae affect adult B. malayi worms and Mf in vitro. Additional studies will be needed to elucidate the mechanisms of action of these antibiotics on Wolbachia and filarial worms.}, } @article {pmid12097909, year = {2002}, author = {Starr, DJ and Cline, TW}, title = {A host parasite interaction rescues Drosophila oogenesis defects.}, journal = {Nature}, volume = {418}, number = {6893}, pages = {76-79}, doi = {10.1038/nature00843}, pmid = {12097909}, issn = {0028-0836}, mesh = {Alleles ; Animals ; *Drosophila Proteins ; Drosophila melanogaster/cytology/genetics/*parasitology/*physiology ; Female ; Gene Expression ; Genes, Lethal/genetics ; Genetic Linkage/genetics ; Germ-Line Mutation/genetics ; Host-Parasite Interactions/physiology ; Male ; Oogenesis/genetics/*physiology ; Ovary/abnormalities/cytology/*physiology ; Phenotype ; RNA-Binding Proteins/genetics/*physiology ; Sex Characteristics ; Sex Determination Processes ; Wolbachia/*physiology ; }, abstract = {The cytoplasmically inherited bacterium Wolbachia pipientis is a widespread parasite of arthropods that manipulates the reproductive biology of its hosts, often to their detriment, in order to foster its own transmission through egg cytoplasm. Here we report that infection by Wolbachia restores fertility to Drosophila melanogaster mutant females prevented from making eggs by protein-coding lesions in Sex-lethal (Sxl), the master regulator of sex determination. Suppression of sterility by Wolbachia discriminates markedly among similar germline-specific Sxl alleles, and is not observed for mutations in other genes that produce similar 'tumorous ovary' phenotypes, including one that blocks Sxl germline expression. This allele and gene specificity indicates that suppression probably results from a specific interaction with Sxl protein, rather than from a bypass of the normal germline requirement for this developmental regulator or from an effect on Sxl expression. The Sxl-Wolbachia interaction provides a rare opportunity to explore host-parasite relationships at the molecular level in a model insect. Furthermore, demonstration that a parasite infection can counteract the deleterious effects of mutations in host genes illustrates how hosts might become dependent on parasites.}, } @article {pmid12095241, year = {2002}, author = {Puttaraju, HP and Madhu, M}, title = {Presence of Wolbachia endosymbionts in different silkworm species and races and in their uzi fly parasites.}, journal = {Journal of invertebrate pathology}, volume = {79}, number = {2}, pages = {120-122}, doi = {10.1016/s0022-2011(02)00031-9}, pmid = {12095241}, issn = {0022-2011}, mesh = {Animals ; Bombyx/classification/*microbiology/*parasitology ; Diptera/*microbiology ; *Ectoparasitic Infestations ; *Symbiosis ; Wolbachia/*isolation & purification ; }, } @article {pmid12095235, year = {2002}, author = {Sokolova, MI and Zinkevich, NS and Zakharov, IA}, title = {Bacteria in ovarioles of females from maleless families of ladybird beetles Adalia bipunctata L. (Coleoptera: Coccinellidae) naturally infected with Rickettsia, Wolbachia, and Spiroplasma.}, journal = {Journal of invertebrate pathology}, volume = {79}, number = {2}, pages = {72-79}, doi = {10.1016/s0022-2011(02)00012-5}, pmid = {12095235}, issn = {0022-2011}, mesh = {Animals ; Bacterial Infections/*microbiology ; Coleoptera/*microbiology ; Rickettsia/*isolation & purification ; Rickettsia Infections/*microbiology ; Spiroplasma/*isolation & purification ; Wolbachia/*isolation & purification ; }, abstract = {Ovarioles were found to be infected with Spiroplasma, Wolbachia, and Rickettsia in Adalia bipunctata females with maleless progeny in different natural populations. Ooplasm was infected with few Wolbachia bacteria. In ooplasm infected by Rickettsia, bacteria were present in small foci. Spiroplasmas were found encapsulated into ooplasm from the wider intercellular spaces between epithelial and oocyte cells. The cytoplasm of follicular epithelia infected with Rickettsia was heavily destroyed, but the nucleus was intact and free from bacteria. The essential feature of follicular epithelium cells from Spiroplasma and Wolbachia infected A. bipunctata females was inclusions of three types: crystalline, filaments, and concentric myelin-like lamellae. Observations of smears prepared from ovaries of A. bipunctata from natural populations revealed a low concentration of bacteria within a microscopy field (less 10 bacteria) in more than 90% of specimens, and only a few ovaries were heavily infected. Two different ways of bacterial invasion of the oocyte are suggested: Spiroplasma-like, through the intercellular spaces in the epithelium and Rickettsia-like, through the cytoplasm of follicular epithelium cells. Bacteria were not found in germarium zones and we suggest that each follicle is infected from haemolymph.}, } @article {pmid12090719, year = {2002}, author = {Hoerauf, A and Adjei, O and Büttner, DW}, title = {Antibiotics for the treatment of onchocerciasis and other filarial infections.}, journal = {Current opinion in investigational drugs (London, England : 2000)}, volume = {3}, number = {4}, pages = {533-537}, pmid = {12090719}, issn = {1472-4472}, mesh = {Animals ; Brugia/microbiology ; Doxycycline/administration & dosage/therapeutic use ; Drug Resistance ; Filariasis/*drug therapy/parasitology ; Filaricides/*therapeutic use ; Humans ; Onchocerca/microbiology ; Onchocerciasis/drug therapy/parasitology ; Symbiosis ; Wolbachia/drug effects ; Wuchereria bancrofti/microbiology ; }, abstract = {More effective drugs are needed for the treatment of human filarial diseases and the elimination of these infections as a public health problem. The drugs must either kill or sterilize adult worms. The relevant filariae, Onchocerca volvulus, Wuchereria bancofti and Brugia species, harbor rickettsial endoboacteria of the genus Wolbachia as symbionts. Animal experiments have shown that the elimination of these endobacteria causes inhibition of embryogenesis, and with Onchocerca ochengi a macrofilaricidal effect. Trials with human onchocerciasis patients using doxycydine demonstrated a long-term sterilizing activity, opening up a new strategy for the control of filarial infections. Indications of antiwolbachial therapy against onchocerciasis are discussed.}, } @article {pmid12079666, year = {2002}, author = {Berticat, C and Rousset, F and Raymond, M and Berthomieu, A and Weill, M}, title = {High Wolbachia density in insecticide-resistant mosquitoes.}, journal = {Proceedings. Biological sciences}, volume = {269}, number = {1498}, pages = {1413-1416}, pmid = {12079666}, issn = {0962-8452}, mesh = {Animals ; Colony Count, Microbial ; Culex/*drug effects/genetics/*microbiology ; Genes, Insect/genetics ; *Insecticide Resistance/genetics ; Insecticides/*pharmacology ; Polymerase Chain Reaction ; Symbiosis ; Wolbachia/genetics/growth & development/*isolation & purification ; }, abstract = {Wolbachia symbionts are responsible for various alterations in host reproduction. The effects of the host genome on endosymbiont levels have often been suggested, but rarely described. Here, we show that Wolbachia density is strongly modified by the presence of insecticide-resistant genes in the common house mosquito, Culex pipiens. The Wolbachia density was estimated using a real-time quantitative PCR assay. Strains harbouring different genes conferring resistance were more infected than a susceptible strain with the same genetic background. We show that this interaction also operates in natural populations. We propose that mosquitoes may control Wolbachia density less efficiently when they carry an insecticide-resistant gene, i.e. when they suffer from a physiological resistance cost.}, } @article {pmid12075374, year = {2002}, author = {Walgate, R}, title = {Could antibiotics cure river blindness?.}, journal = {Bulletin of the World Health Organization}, volume = {80}, number = {4}, pages = {336}, pmid = {12075374}, issn = {0042-9686}, mesh = {Africa ; Animals ; Causality ; Doxycycline/adverse effects/pharmacology/therapeutic use ; Humans ; Onchocerca volvulus/drug effects/*microbiology ; Onchocerciasis, Ocular/*drug therapy/microbiology/parasitology ; Tetracyclines/adverse effects/pharmacology/*therapeutic use ; Wolbachia/drug effects/*pathogenicity ; }, } @article {pmid12062980, year = {2002}, author = {Habeck, M}, title = {Tracking down the culprit in river blindness.}, journal = {The Lancet. Infectious diseases}, volume = {2}, number = {5}, pages = {262}, doi = {10.1016/s1473-3099(02)00274-8}, pmid = {12062980}, issn = {1473-3099}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Doxycycline/therapeutic use ; Humans ; Onchocerca/drug effects/*microbiology ; Onchocerciasis, Ocular/drug therapy/*microbiology ; Wolbachia/*pathogenicity ; }, } @article {pmid12061400, year = {2002}, author = {Hiroki, M and Kato, Y and Kamito, T and Miura, K}, title = {Feminization of genetic males by a symbiotic bacterium in a butterfly, Eurema hecabe (Lepidoptera: Pieridae).}, journal = {Die Naturwissenschaften}, volume = {89}, number = {4}, pages = {167-170}, doi = {10.1007/s00114-002-0303-5}, pmid = {12061400}, issn = {0028-1042}, mesh = {Animals ; Bacterial Proteins/genetics ; Butterflies/drug effects/*physiology ; *Cytoskeletal Proteins ; Female ; *Feminization ; Larva ; Male ; Oviposition ; Reproduction/*physiology ; Rickettsiaceae/*physiology ; Sex Ratio ; Symbiosis ; Tetracycline/pharmacology ; }, abstract = {Wolbachia are symbiotic bacteria found in many arthropods and filarian nematodes. They often manipulate the reproduction of host arthropods. In the present study, female-biased sex-ratio distortion in the butterfly Eurema hecabe was investigated. Breeding experiments showed that this distorted sex ratio is maternally inherited. When treated with tetracycline, adult females of the thelygenic line produced male progeny only. After PCR using Wolbachia-specific primers for the ftsZ gene a positive result was seen in the thelygenic females, but not in male progeny from tetracycline-treated females, or individuals from a Tokyo population with normal sex ratio and reproduction. Cytological observations showed that thelygenic females lack the sex chromatin body (W chromosome). The results strongly suggest that the sex-ratio distortion in E. hecabe is due to feminization of genetic males by Wolbachia.}, } @article {pmid12051989, year = {2002}, author = {Schofield, P}, title = {Spatially explicit models of Turelli-Hoffmann Wolbachia invasive wave fronts.}, journal = {Journal of theoretical biology}, volume = {215}, number = {1}, pages = {121-131}, doi = {10.1006/jtbi.2001.2493}, pmid = {12051989}, issn = {0022-5193}, mesh = {Animals ; Drosophila/*microbiology ; Female ; Insect Vectors/*microbiology ; Male ; *Models, Biological ; Rickettsiaceae Infections/*transmission ; *Wolbachia ; }, abstract = {This paper examines different mathematical models of insect dispersal and infection spread and compares these with field data. Reaction-diffusion and integro-difference equation models are used to model the spatio-temporal spread of Wolbachia in Drosophila simulans populations. The models include cytoplasmic incompatibility between infected females and uninfected males that creates a threshold density, similar to an Allee effect, preventing increase from low incidence of infection in the host population. The model builds on an earlier model (Turelli & Hoffmann, 1991) by incorporating imperfect maternal transmission. The results of simulations of the models using the same parameter values produce different dynamics for each model. These differences become very marked in the integro-difference equation models when insect dispersal patterns are assumed to be non-Gaussian. The success or failure of invasion by Wolbachia in the simulations may be attributed to the insect dispersal mechanism used in the model rather than the parameter values. As the models predict very different outcomes for the integro-difference models depending on the underlying assumptions of insect dispersal patterns, this emphasizes that good field data on real (rather than idealized) dispersal patterns need to be collected before models such as these can be used for predictive purposes.}, } @article {pmid12041732, year = {2002}, author = {Taylor, MJ}, title = {A new insight into the pathogenesis of filarial disease.}, journal = {Current molecular medicine}, volume = {2}, number = {3}, pages = {299-302}, doi = {10.2174/1566524024605662}, pmid = {12041732}, issn = {1566-5240}, mesh = {Animals ; Filariasis/*etiology/parasitology/pathology ; Humans ; Wolbachia/pathogenicity ; }, abstract = {Filariasis is a major public health problem throughout many regions of the tropics. The disease is caused by several species of filarial nematode including Wuchereria bancrofti and Brugia malayi, the agents of lymphatic filariasis, and Onchocerca volvulus, the cause of 'riverblindness'. Disease caused by these worms varies depending on the tissue location of the parasite, and is associated with episodes of acute and chronic inflammation. These pathologies, including elephantiasis and blindness, rank among the most disabling in the world. Studies aimed at characterizing the molecular nature of the inflammatory stimuli derived from filarial nematodes uncovered a long forgotten secret, their symbiont Wolbachia. LPS-like molecules from these intracellular bacteria are responsible for potent inflammatory responses from macrophages and in animal models of filarial disease. Wolbachia has also been associated with severe inflammatory reactions to filarial chemotherapy, being released into the blood following the death of the parasite. Recent studies in animal models even implicate Wolbachia in the onset of lymphodema and blindness. Taken together these studies suggest a major role for Wolbachia in the pathogenesis of filarial disease. It may be possible, through the use of antibiotic therapy, to clear worms of their bacteria, in the hope that this will prevent the onset and development of filarial pathology.}, } @article {pmid12029099, year = {2002}, author = {Kennedy, D}, title = {Good news on a tropical disease.}, journal = {Science (New York, N.Y.)}, volume = {296}, number = {5572}, pages = {1365}, doi = {10.1126/science.296.5572.1365}, pmid = {12029099}, issn = {1095-9203}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/*pharmacology/therapeutic use ; Developing Countries ; Filaricides/administration & dosage/therapeutic use ; Humans ; Ivermectin/administration & dosage/*therapeutic use ; Onchocerca volvulus/drug effects/microbiology/*physiology ; Onchocerciasis, Ocular/epidemiology/parasitology/*prevention & control/transmission ; Organizations, Nonprofit ; Symbiosis ; Wolbachia/drug effects/*physiology ; World Health Organization ; }, } @article {pmid12009809, year = {2001}, author = {Tagami, Y and Miura, K and Stouthamer, R}, title = {How does infection with parthenogenesis-inducing Wolbachia reduce the fitness of Trichogramma?.}, journal = {Journal of invertebrate pathology}, volume = {78}, number = {4}, pages = {267-271}, doi = {10.1006/jipa.2002.5080}, pmid = {12009809}, issn = {0022-2011}, mesh = {Animals ; Female ; Wasps/embryology/*microbiology ; Wolbachia/*physiology ; }, abstract = {We analyzed the survival rate of the immature stages of Trichogramma species and lines that differed in their mode of reproduction. Specifically, we compared the mortality of arrhenotokous (W(-)), irrevertable thelytokous (W(-)), and Wolbachia-associated thelytokous (W(+)) forms. The embryonic mortality of the W(+) strains was significantly higher than that of the W(-) lines. The embryonic mortality was negligible for the arrhenotokous Trichogramma evanescens and the thelytokous Trichogramma cacoeciae, which is not infected with Wolbachia. Only 30% of the eggs of the Wolbachia-infected strains developed to adults, while the emergence rate of the Wolbachia-free strains was more than 78%, irrespective of the origin of the strains. More than 78% of the overall mortality in W(+) strains happened during the early stages of development. About 35% of embryos of W(+) strains remain in the mitotic stage even 48 h after oviposition. Most embryos of W(-) strains had already developed to cellular blastoderm after 6 h, regardless of strains. The mortality of immature stages in W(+) strains was mainly caused by the failure of the mitotic divisions.}, } @article {pmid12004132, year = {2002}, author = {Tram, U and Sullivan, W}, title = {Role of delayed nuclear envelope breakdown and mitosis in Wolbachia-induced cytoplasmic incompatibility.}, journal = {Science (New York, N.Y.)}, volume = {296}, number = {5570}, pages = {1124-1126}, doi = {10.1126/science.1070536}, pmid = {12004132}, issn = {1095-9203}, support = {GM16409/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; CDC2 Protein Kinase/metabolism ; Centrosome/physiology ; Chromosomes/physiology ; Cytoplasm/physiology ; Embryo, Nonmammalian/physiology ; Female ; Fertilization ; Histones/metabolism ; Male ; *Mitosis ; Motion Pictures ; Nuclear Envelope/*physiology ; Ovum/microbiology/physiology ; Phosphorylation ; Spermatozoa/microbiology/physiology ; Spindle Apparatus/physiology ; Time Factors ; Wasps/embryology/*microbiology/*physiology ; Wolbachia/*physiology ; Zygote/*physiology ; }, abstract = {The bacterium Wolbachia manipulates reproduction in millions of insects worldwide; the most common effect is cytoplasmic incompatibility (CI). We found that CI resulted from delayed nuclear envelope breakdown of the male pronucleus in Nasonia vitripennis. This caused asynchrony between the male and female pronuclei and, ultimately, loss of paternal chromosomes at the first mitosis. When Wolbachia were present in the egg, synchrony was restored, which explains suppression of CI in these crosses. These results suggest that Wolbachia target cell cycle regulatory proteins. A striking consequence of CI is that it alters the normal pattern of reciprocal centrosome inheritance in Nasonia.}, } @article {pmid12004092, year = {2002}, author = {Zimmer, C}, title = {Evolutionary biology. Timing is everything for Wolbachia hosts.}, journal = {Science (New York, N.Y.)}, volume = {296}, number = {5570}, pages = {999-1000}, doi = {10.1126/science.296.5570.999a}, pmid = {12004092}, issn = {1095-9203}, mesh = {Animals ; *Biological Evolution ; Cell Division ; Chromosomes/physiology ; Embryo, Nonmammalian/physiology ; Female ; Fertilization ; Male ; Motion Pictures ; Nuclear Envelope/physiology ; Reproduction ; Sex Determination Processes ; Sex Ratio ; Time Factors ; Wasps/embryology/*microbiology/*physiology ; Wolbachia/*physiology ; Zygote/*physiology ; }, } @article {pmid11992713, year = {2002}, author = {Egyed, Z and Sréter, T and Széll, Z and Nyirö, G and Dobos-Kovács, M and Márialigeti, K and Varga, I}, title = {Electron microscopic and molecular identification of Wolbachia endosymbionts from Onchocerca lupi: implications for therapy.}, journal = {Veterinary parasitology}, volume = {106}, number = {1}, pages = {75-82}, doi = {10.1016/s0304-4017(02)00029-8}, pmid = {11992713}, issn = {0304-4017}, mesh = {Animals ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Dog Diseases/drug therapy/*parasitology ; Dogs ; Female ; Male ; Microscopy, Electron ; Onchocerca/genetics/growth & development/*microbiology ; Onchocerciasis, Ocular/drug therapy/parasitology/*veterinary ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Homology, Nucleic Acid ; Symbiosis ; Wolbachia/*genetics/growth & development/ultrastructure ; }, abstract = {It was recently demonstrated that Wolbachia intracellular bacteria (alpha 2 proteobacteria, Rickettsiales) living in filarial nematodes are obligatory symbionts of their hosts. Herein, we report the electron microscopic and 16S ribosomal DNA-based (16S rDNA) identification of the endobacteria harboring in Onchocerca lupi. The worm nodules containing the nematodes were removed from three Hungarian dogs naturally infected with O. lupi. Wolbachia-like endobacteria were detected by electron microscopy in the lateral chords of both adult worms and microfilariae. The endosymbionts in O. lupi resemble in location, size, and morphology the wolbachiae found in other filariae. The presence of wolbachiae in O. lupi was also confirmed by PCR amplification of the 16S rDNA of the bacteria. The 16S rDNA-based phylogenetic analysis revealed that the endosymbionts of O. lupi infecting dogs belong to the supergroup C of Wolbachia pipientis and are not identical with those of other Onchocerca spp. sequenced so far. Since intermittent treatment with oxytetracycline has adulticid and microfilaricid activity by depletion of Wolbachia endobacteria, this antibiotic treatment regimen may offer an alternative of ivermectin or diethylcarbamazine in the suppression of postoperative microfilaridermia in Onchocerca-infected dogs and may prevent relapse.}, } @article {pmid11989683, year = {2002}, author = {Ballard, JW and Chernoff, B and James, AC}, title = {Divergence of mitochondrial dna is not corroborated by nuclear dna, morphology, or behavior in Drosophila simulans.}, journal = {Evolution; international journal of organic evolution}, volume = {56}, number = {3}, pages = {527-545}, doi = {10.1111/j.0014-3820.2002.tb01364.x}, pmid = {11989683}, issn = {0014-3820}, mesh = {Animals ; Base Sequence ; Behavior, Animal/*physiology ; Cell Nucleus/*genetics ; DNA/*genetics ; DNA Primers ; DNA, Mitochondrial/*genetics ; Drosophila/classification/*genetics/physiology ; *Evolution, Molecular ; Female ; Genetic Variation ; Geography ; *Phylogeny ; Reproduction/physiology ; Wolbachia/classification/genetics/physiology ; X Chromosome ; }, abstract = {We ask whether the observed mitochondrial DNA (mtDNA) population subdivision of Drosophila simulans is indicative of organismal structure or of specific processes acting on the mitochondrial genome. Factors either intrinsic or extrinsic to the host genome may influence the evolutionary dynamics of mtDNA. Potential intrinsic factors include adaptation of the mitochondrial genome and of nucleomitochondrial gene complexes specific to the local environment. An extrinsic force that has been shown to influence mtDNA evolution in invertebrates is the bacterial endosymbiont Wolbachia. Evidence presented in this study suggests that mtDNA is not a good indicator of organismal subdivision in D. simulans. Furthermore, there is no evidence to suggest that Wolbachia causes any reduction in nuclear gene flow in this species. The observed differentiation in mtDNA is not corroborated by data from NADH: ubiquinone reductase 75kD subunit precursor or the Alcohol dehydrogenase-related loci, from the shape or size of the male genital arch, or from assortative premating behavior. We discuss these results in relation to a mitochondrial genetic species concept and the potential for Wolbachia-induced incompatibility to be a mechanism of speciation in insects. We conclude with an iterated appeal to include phylogenetic and statistical tests of neutrality as a supplement to phylogenetic and population genetic analyses when using mtDNA as an evolutionary marker.}, } @article {pmid11986872, year = {2002}, author = {Vavre, F and Fleury, F and Varaldi, J and Fouillet, P and Boulétreau, M}, title = {Infection polymorphism and cytoplasmic incompatibility in Hymenoptera-Wolbachia associations.}, journal = {Heredity}, volume = {88}, number = {5}, pages = {361-365}, doi = {10.1038/sj.hdy.6800063}, pmid = {11986872}, issn = {0018-067X}, mesh = {Animals ; Cytoplasm/physiology ; Drosophila/parasitology ; Female ; Fertility ; Male ; Polymorphism, Genetic ; Reproduction ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {Most cases of Wolbachia infection so far documented in haplodiploid Hymenoptera are associated with parthenogenesis induction. Only three examples of Wolbachia-mediated cytoplasmic incompatibility (CI) have been reported, resulting either in haploidisation of fertilised eggs, which develop into viable males, or in their death. To better document this variability, we studied two new Wolbachia-wasp associations involving Drosophila parasitoids. In Trichopria cf. drosophilae, individuals are infected by two different Wolbachia variants, populations are nearly totally infected, and Wolbachia induces incomplete CI resulting in death of the fertilised eggs. On the other hand, Pachycrepoideus dubius harbours only one bacterial variant, populations are polymorphic for infection, and Wolbachia has no detectable effect. These two cases show that the range of variation in Wolbachia's effects in Hymenoptera is as wide as in diploids, extending from complete CI to an undetectable effect. Cases so far studied show some parallel between the strength of incompatibility, the number of Wolbachia variants infecting each wasp, and the natural infection frequency. These empirical data support theoretical models predicting evolution of CI towards lower levels, resulting in the decline and ultimate loss of infection, and place multiple infections as being an important factor in the evolution of host-Wolbachia associations.}, } @article {pmid11965304, year = {2002}, author = {Blanke, CH}, title = {Wolbachia DNA recognition.}, journal = {Lancet (London, England)}, volume = {359}, number = {9314}, pages = {1345-1346}, doi = {10.1016/s0140-6736(02)08292-2}, pmid = {11965304}, issn = {0140-6736}, mesh = {Antigens, Bacterial/*immunology ; DNA, Bacterial/*immunology ; Filariasis/drug therapy/*immunology ; Filaricides/therapeutic use ; Humans ; Ivermectin/therapeutic use ; Species Specificity ; Wolbachia/*immunology ; }, } @article {pmid11964892, year = {2001}, author = {Taylor, MJ and Hoerauf, A}, title = {A new approach to the treatment of filariasis.}, journal = {Current opinion in infectious diseases}, volume = {14}, number = {6}, pages = {727-731}, doi = {10.1097/00001432-200112000-00011}, pmid = {11964892}, issn = {0951-7375}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Disease Models, Animal ; Filariasis/*drug therapy/microbiology ; Humans ; Symbiosis ; Wolbachia/*drug effects ; }, abstract = {The symbiosis of filarial nematodes and intracellular Wolbachia bacteria has recently been exploited as a target for antibiotic therapy of filariasis. Antibiotic treatment of filarial nematodes results in sterility and inhibits larval development and adult worm viability. In the first trial on human onchocerciasis depletion of bacteria following treatment with doxycycline resulted in a complete and long-term block of embryogenesis. Bacteria are unable to repopulate nematode tissues up to 18 months after depletion, suggesting these effects may be permanent. Following ivermectin treatment, individuals given antibiotic therapy showed sustained reductions in skin microfilariae, with the majority of people remaining microfilarial negative 12-18 months after treatment. Since Wolbachia also contribute to the inflammatory pathogenesis of filarial disease, antibiotic therapy could, in addition to effects on worm fertility or viability, prevent the onset or development of filarial pathology.}, } @article {pmid11964873, year = {2001}, author = {Nutman, TB}, title = {Lymphatic filariasis: new insights and prospects for control.}, journal = {Current opinion in infectious diseases}, volume = {14}, number = {5}, pages = {539-546}, doi = {10.1097/00001432-200110000-00006}, pmid = {11964873}, issn = {0951-7375}, mesh = {Animals ; Elephantiasis, Filarial/diagnosis/*prevention & control/therapy ; Humans ; Wuchereria bancrofti/genetics/immunology/pathogenicity ; }, abstract = {Although lymphatic filariasis remains among the major causes of disability among the tropical infectious diseases, dramatic advances have been made in the approach to its diagnosis, epidemiology and treatment, in our understanding of the molecular composition of the parasites that cause these infections, and in the factors underlying the pathology seen. Superimposing the tools of modern epidemiology, immunology, and molecular biology on field-based clinical trials has allowed the emergence of the concept of elimination of lymphatic filariasis. Much of the important new research emphasizes parasite development in the context of the host response, the importance of both the adult worm and other factors in the pathogenesis of lymphatic filarial disease, the role the Wolbachia endosymbiont holds as both a target for drug treatment and in inducing post-treatment reactions, and the various principles underlying the implementation of control programs.}, } @article {pmid11964814, year = {2000}, author = {Mackenzie, CD}, title = {Human onchocerciasis: the essential partnership between research and disease control efforts.}, journal = {Current opinion in infectious diseases}, volume = {13}, number = {5}, pages = {457-464}, doi = {10.1097/00001432-200010000-00005}, pmid = {11964814}, issn = {1473-6527}, abstract = {Twenty years ago onchocerciasis was a disease generally ignored by the medical world, except by those who actually worked with the affected people in Africa and Latin America. Now, largely as a result of the success of mass vector control and drug treatment programs, this is a disease management model for developing countries. The recent literature on onchocerciasis has, not surprisingly, mainly focused on various aspects of control. Investigation into the more basic questions is needed to ensure continued effective disease control. The present mass drug control program is based on a single pharmaceutical, ivermectin (Mectizan), which acts almost exclusively on the microfilarial stage of the infection. Efforts are being made to identify other useful drugs; however, no major candidates have yet appeared. The identification of potential biochemical targets for anti-filarial compounds through a better understanding of the biochemistry of these worms is being pursued. The Onchocerca volvulus endosymbiont Wolbachia may provide a target for therapeutic intervention. An improved understanding of the genomics of O. volvulus has made possible the identification of strain differences in the parasites, and an appreciation of the relevance of these strain differences to the clinical disease, onchocerciasis. There is a need for a better understanding of the clinical disease, and the various pathogenic mechanisms that underly the different syndromes. It is particularly important to understand the pathological basis and mechanisms underlying the adverse responses that can occur with chemotherapy. Present control programs now need to be carefully monitored for effectiveness using new assessment tools, such as antigen assays and the identification of organisms in pools of vectors. Current efforts to control onchocerciasis must be coordinated with new chemotherapy-based control programs for other worm diseases that are emerging. The results of laboratory studies are increasingly being applied to improve the effectiveness of field-based control programs and their assessment. Such research is essential for progress towards the goals of controlling and eliminating onchocerciasis.}, } @article {pmid11925998, year = {2002}, author = {Blaxter, M and Daub, J and Guiliano, D and Parkinson, J and Whitton, C and , }, title = {The Brugia malayi genome project: expressed sequence tags and gene discovery.}, journal = {Transactions of the Royal Society of Tropical Medicine and Hygiene}, volume = {96}, number = {1}, pages = {7-17}, doi = {10.1016/s0035-9203(02)90224-5}, pmid = {11925998}, issn = {0035-9203}, mesh = {Animals ; Brugia malayi/*genetics/parasitology ; Conserved Sequence ; *Expressed Sequence Tags ; Genome, Bacterial ; *Genome, Protozoan ; Genomic Library ; Symbiosis ; Wolbachia/genetics ; }, abstract = {To advance and facilitate molecular studies of Brugia malayi, one of the causative agents of human lymphatic filariasis, an expressed sequence tag (EST)-based gene discovery programme has been carried out. Over 22,000 ESTs have been produced and deposited in the public databases by a consortium of laboratories from endemic and non-endemic countries. The ESTs have been analysed using custom informatic tools to reveal patterns of individual gene expression that may point to potential targets for future research on anti-filarial drugs and vaccines. Many genes first discovered as ESTs are now being analysed by researchers for immunodiagnostic, vaccine and drug target potential. Building on the success of the B. malayi EST programme, significant EST datasets are being generated for a number of other major parasites of humans and domesticated animals, and model parasitic species.}, } @article {pmid11920298, year = {2002}, author = {Keiser, PB and Reynolds, SM and Awadzi, K and Ottesen, EA and Taylor, MJ and Nutman, TB}, title = {Bacterial endosymbionts of Onchocerca volvulus in the pathogenesis of posttreatment reactions.}, journal = {The Journal of infectious diseases}, volume = {185}, number = {6}, pages = {805-811}, doi = {10.1086/339344}, pmid = {11920298}, issn = {0022-1899}, mesh = {Adolescent ; Adult ; Calcium-Binding Proteins/blood ; Calgranulin B ; DNA, Bacterial/analysis ; Diethylcarbamazine/*adverse effects ; Filaricides/*adverse effects ; Humans ; Ivermectin/*adverse effects ; Leukocyte L1 Antigen Complex ; Male ; Membrane Glycoproteins/blood ; Neural Cell Adhesion Molecules/blood ; Onchocerciasis/*drug therapy/immunology ; *Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {Treatment of onchocerciasis with diethylcarbamazine (DEC) or ivermectin is associated with a posttreatment reaction characterized by fever, tachycardia, hypotension, lymphadenopathy, and pruritus. To investigate the role of the Wolbachia bacterial endosymbiont of Onchocerca volvulus in these reactions, serum samples collected before and after treatment with either anthelmintic were assessed for evidence of Wolbachia DNA. By use of real-time quantitative polymerase chain reaction, Wolbachia DNA was detected in both groups-with significantly higher levels in those who received DEC (P <.0001). In the ivermectin group, there was a significant correlation between levels of bacterial DNA and serum tumor necrosis factor-alpha (P =.013). Peak DNA levels correlated with reaction scores (P =.048). Significant correlations were also seen between Wolbachia DNA and the antibacterial peptides calprotectin (P =.021) and calgranulin B (P <.0001). These findings support a role for Wolbachia products in mediating the inflammatory responses seen following treatment of onchocerciasis and suggest new targets for modulating these reactions.}, } @article {pmid11920134, year = {2002}, author = {Kittayapong, P and Mongkalangoon, P and Baimai, V and O'Neill, SL}, title = {Host age effect and expression of cytoplasmic incompatibility in field populations of Wolbachia-superinfected Aedes albopictus.}, journal = {Heredity}, volume = {88}, number = {4}, pages = {270-274}, doi = {10.1038/sj.hdy.6800039}, pmid = {11920134}, issn = {0018-067X}, mesh = {Aedes/*microbiology/physiology ; Age Factors ; Animals ; Crosses, Genetic ; Cytoplasm/*physiology ; DNA/metabolism ; Female ; Male ; Polymerase Chain Reaction ; Wolbachia/*physiology ; }, abstract = {The Asian tiger mosquito, Aedes albopictus (Skuse), is a known vector of dengue in South America and Southeast Asia. It is naturally superinfected with two strains of Wolbachia endosymbiont that are able to induce cytoplasmic incompatibility (CI). In this paper, we report the strength of CI expression in crosses involving field-caught males. CI expression was found to be very strong in all crosses between field males and laboratory-reared uninfected or wAlbA infected young females. In addition, crossing experiments with laboratory colonies showed that aged super-infected males could express strong CI when mated with young uninfected or wAlbA infected females. These results provide additional evidence that the CI properties of Wolbachia infecting Aedes albopictus are well suited for applied strategies that seek to utilise Wolbachia for host population modification.}, } @article {pmid11920120, year = {2002}, author = {Hong, XY and Gotoh, T and Nagata, T}, title = {Vertical transmission of Wolbachia in Tetranychus kanzawai Kishida and Panonychus mori Yokoyama (Acari: Tetranychidae).}, journal = {Heredity}, volume = {88}, number = {3}, pages = {190-196}, doi = {10.1038/sj.hdy.6800026}, pmid = {11920120}, issn = {0018-067X}, mesh = {Animals ; DNA/metabolism ; Female ; *Gram-Negative Bacterial Infections ; Infectious Disease Transmission, Vertical ; Male ; Mites/*microbiology ; Models, Statistical ; Reproduction/*physiology ; Sexual Behavior, Animal ; Species Specificity ; Time Factors ; Wolbachia/*metabolism ; }, abstract = {The vertical transmission of Wolbachia in two species of spider mite was investigated and compared. One species, Tetranychus kanzawai Kishida, was infected with a modification negative strain of Wolbachia while the other species, Panonychus mori Yokoyama, was infected with a modification positive strain. The infection showed perfect maternal transmission in the laboratory population of T. kanzawai in which Wolbachia-infected females produced infected offspring regardless of whether they mated with infected or uninfected males, and uninfected females produced Wolbachia-free progenies without regard to the infection status of their mating partners. In artificial P. mori populations initiated with 50% infected and 50% uninfected female adults, the infection frequencies among progenies increased with each generation, reaching 100% at the sixth generation in the Sendai population and after the sixth generation in the Toyama population. In another experiment, in which an artificial T. kanzawai population was composed of 50% infected and 50% uninfected female adults, the infection frequency in progeny populations increased very slowly, reaching 62.5% at the 15th generation. The difference in infection frequency in the two spider mites may be due to the different strains of Wolbachia.}, } @article {pmid11920119, year = {2002}, author = {James, AC and Dean, MD and McMahon, ME and Ballard, JW}, title = {Dynamics of double and single Wolbachia infections in Drosophila simulans from New Caledonia.}, journal = {Heredity}, volume = {88}, number = {3}, pages = {182-189}, doi = {10.1038/sj.hdy.6800025}, pmid = {11920119}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm/metabolism ; DNA, Mitochondrial/metabolism ; Drosophila/metabolism ; Female ; Gram-Negative Bacterial Infections/*microbiology/*pathology ; Male ; Models, Genetic ; New Caledonia ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sex Ratio ; Sexual Behavior, Animal ; Wolbachia/*metabolism ; }, abstract = {The bacterial symbiont Wolbachia can cause cytoplasmic incompatibility in Drosophila simulans flies: if an infected male mates with an uninfected female, or a female with a different strain of Wolbachia, there can be a dramatic reduction in the number of viable eggs produced. Here we explore the dynamics associated with double and single Wolbachia infections in New Caledonia. Doubly infected females were compatible with all males in the population, explaining the high proportion of doubly infected flies. In this study, males that carry only wHa or wNo infections showed reduced incompatibility when mated to uninfected females, compared with previous reports. These data suggest that either the DNA of these bacterial isolates have diverged from those previously collected, or the genetic background of the host has lead to a reduction in the phenotype of incompatibility. Mitochondrial sequence polymorphism at two sites within the host genome was assayed to investigate population structure related to infection types. There was no correlation between sequence polymorphism and infection type suggesting that double infections are the stable type, with singly infected and uninfected flies arising from stochastic segregation of bacterial strains. Finally, we discuss the nomenclature of Wolbachia strain designation.}, } @article {pmid11920117, year = {2002}, author = {Dyson, EA and Kamath, MK and Hurst, GD}, title = {Wolbachia infection associated with all-female broods in Hypolimnas bolina (Lepidoptera: Nymphalidae): evidence for horizontal transmission of a butterfly male killer.}, journal = {Heredity}, volume = {88}, number = {3}, pages = {166-171}, doi = {10.1038/sj.hdy.6800021}, pmid = {11920117}, issn = {0018-067X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; Crosses, Genetic ; *Cytoskeletal Proteins ; Female ; Gram-Negative Bacterial Infections/*microbiology ; Lepidoptera/microbiology/*physiology ; Male ; Phylogeny ; Sequence Analysis, DNA ; Sex Ratio ; Sexual Behavior, Animal ; Wolbachia/*metabolism/physiology ; }, abstract = {Inherited bacteria that kill male hosts during embryogenesis infect a wide range of insect species. In order to ascertain if there are patterns to host infection, with particular male killing bacteria specialising on particular taxa, we investigated the male killing trait in the butterfly Hypolimnas bolina. All-female broods were first reported in this species in the 1920s. Investigation of this system in the Fiji Islands revealed the causal agent of sex ratio distortion in H. bolina to be a male killing Wolbachia bacterium. This bacterium is identical in wsp and ftsZ sequence to a male killer in the butterfly Acraea encedon in Tanzania, suggesting it has moved between host species, yet retained its phenotype. The prevalence of the Wolbachia was calculated for three different island groups of Fiji, and found to vary significantly across the country. Antibiotics failed to cure either the male killing trait or the Wolbachia infection. The implications of these results are discussed.}, } @article {pmid11916479, year = {2002}, author = {Wenseleers, T and Sundström, L and Billen, J}, title = {Deleterious Wolbachia in the ant Formica truncorum.}, journal = {Proceedings. Biological sciences}, volume = {269}, number = {1491}, pages = {623-629}, pmid = {11916479}, issn = {0962-8452}, mesh = {Animals ; Ants/*microbiology/*physiology ; Biological Evolution ; Female ; Host-Parasite Interactions ; Male ; Molecular Sequence Data ; Phylogeny ; Reproduction/physiology ; Sex Ratio ; Species Specificity ; Wolbachia/genetics/*pathogenicity/physiology ; }, abstract = {Wolbachia is a maternally inherited bacterium that may manipulate the reproduction of its arthropod hosts. In insects, it is known to lead to inviable matings, cause asexual reproduction or kill male offspring, all to its own benefit, but to the detriment of its host. In social Hymenoptera, Wolbachia occurs widely, but little is known about its fitness effects. We report on a Wolbachia infection in the wood ant Formica truncorum, and evaluate whether it influences reproductive patterns. All 33 colonies of the study population were infected, suggesting that Wolbachia infection is at, or close to, fixation. Interestingly, in colonies with fewer infected workers, significantly more sexuals are produced, indicating that Wolbachia has deleterious effects in this species. In addition, adult workers are shown to have significantly lower infection rates (45%) than worker pupae (87%) or virgin queens (94%), suggesting that workers lose their infection over life. Clearance of Wolbachia infection has, to our knowledge, never been shown in any other natural system, but we argue that it may, in this case, represent an adaptive strategy to reduce colony load. The cause of fixation requires further study, but our data strongly suggest that Wolbachia has no influence on the sex ratio in this species.}, } @article {pmid11905109, year = {2002}, author = {Hurst, LD and Randerson, JP}, title = {Parasitic sex puppeteers.}, journal = {Scientific American}, volume = {286}, number = {4}, pages = {56-61}, doi = {10.1038/scientificamerican0402-56}, pmid = {11905109}, issn = {0036-8733}, mesh = {Animals ; Arthropods/genetics/*microbiology/*physiology ; Biological Evolution ; Butterflies/genetics/microbiology/physiology ; Crustacea/genetics/microbiology/physiology ; Drosophila/genetics/microbiology/physiology ; Female ; Genetics, Population ; Male ; Reproduction ; Sex Determination Processes ; Sex Ratio ; Wasps/classification/microbiology/physiology ; Wolbachia/*physiology ; }, } @article {pmid11901124, year = {2002}, author = {Dobson, SL and Marsland, EJ and Rattanadechakul, W}, title = {Mutualistic Wolbachia infection in Aedes albopictus: accelerating cytoplasmic drive.}, journal = {Genetics}, volume = {160}, number = {3}, pages = {1087-1094}, pmid = {11901124}, issn = {0016-6731}, mesh = {Aedes/*microbiology ; Animals ; Biological Evolution ; Female ; Fertility/physiology ; Genetics, Population ; Male ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {Maternally inherited rickettsial symbionts of the genus Wolbachia occur commonly in arthropods, often behaving as reproductive parasites by manipulating host reproduction to enhance the vertical transmission of infections. One manipulation is cytoplasmic incompatibility (CI), which causes a significant reduction in brood hatch and promotes the spread of the maternally inherited Wolbachia infection into the host population (i.e., cytoplasmic drive). Here, we have examined a Wolbachia superinfection in the mosquito Aedes albopictus and found the infection to be associated with both cytoplasmic incompatibility and increased host fecundity. Relative to uninfected females, infected females live longer, produce more eggs, and have higher hatching rates in compatible crosses. A model describing Wolbachia infection dynamics predicts that increased fecundity will accelerate cytoplasmic drive rates. To test this hypothesis, we used population cages to examine the rate at which Wolbachia invades an uninfected Ae. albopictus population. The observed cytoplasmic drive rates were consistent with model predictions for a CI-inducing Wolbachia infection that increases host fecundity. We discuss the relevance of these results to both the evolution of Wolbachia symbioses and proposed applied strategies for the use of Wolbachia infections to drive desired transgenes through natural populations (i.e., population replacement strategies).}, } @article {pmid11901123, year = {2002}, author = {v d Schulenburg, JH and Hurst, GD and Tetzlaff, D and Booth, GE and Zakharov, IA and Majerus, ME}, title = {History of infection with different male-killing bacteria in the two-spot ladybird beetle Adalia bipunctata revealed through mitochondrial DNA sequence analysis.}, journal = {Genetics}, volume = {160}, number = {3}, pages = {1075-1086}, pmid = {11901123}, issn = {0016-6731}, mesh = {Animals ; Biological Evolution ; Coleoptera/*genetics/*microbiology ; *DNA, Mitochondrial ; Electron Transport Complex IV/genetics ; Genotype ; Male ; Molecular Sequence Data ; NADH Dehydrogenase/genetics ; Phylogeny ; Rickettsia/*genetics ; Sequence Analysis, DNA ; Spiroplasma/*genetics ; *Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {The two-spot ladybird beetle Adalia bipunctata (Coleoptera: Coccinellidae) is host to four different intracellular maternally inherited bacteria that kill male hosts during embryogenesis: one each of the genus Rickettsia (alpha-Proteobacteria) and Spiroplasma (Mollicutes) and two distinct strains of Wolbachia (alpha-Proteobacteria). The history of infection with these male-killers was explored using host mitochondrial DNA, which is linked with the bacteria due to joint maternal inheritance. Two variable regions, 610 bp of cytochrome oxidase subunit I and 563 bp of NADH dehydrogenase subunit 5, were isolated from 52 A. bipunctata with known infection status and different geographic origin from across Eurasia. Two outgroup taxa were also considered. DNA sequence analysis revealed that the distribution of mitochondrial haplotypes is not associated with geography. Rather, it correlates with infection status, confirming linkage disequilibrium between mitochondria and bacteria. The data strongly suggest that the Rickettsia male-killer invaded the host earlier than the other taxa. Further, the male-killing Spiroplasma is indicated to have undergone a recent and extensive spread through host populations. In general, male-killing in A. bipunctata seems to represent a highly dynamic system, which should prove useful in future studies on the evolutionary dynamics of this peculiar type of symbiont-host association.}, } @article {pmid11886634, year = {2002}, author = {Dobson, SL and Fox, CW and Jiggins, FM}, title = {The effect of Wolbachia-induced cytoplasmic incompatibility on host population size in natural and manipulated systems.}, journal = {Proceedings. Biological sciences}, volume = {269}, number = {1490}, pages = {437-445}, pmid = {11886634}, issn = {0962-8452}, mesh = {Animals ; Computer Simulation ; Crosses, Genetic ; Female ; Host-Parasite Interactions ; Infectious Disease Transmission, Vertical ; Infertility/genetics/microbiology ; Insecta/*cytology/genetics/*microbiology ; Male ; Models, Biological ; Population Dynamics ; Rickettsiaceae Infections/genetics/microbiology/transmission ; Wolbachia/*physiology ; }, abstract = {Obligate, intracellular bacteria of the genus Wolbachia often behave as reproductive parasites by manipulating host reproduction to enhance their vertical transmission. One of these reproductive manipulations, cytoplasmic incompatibility, causes a reduction in egg-hatch rate in crosses between individuals with differing infections. Applied strategies based upon cytoplasmic incompatibility have been proposed for both the suppression and replacement of host populations. As Wolbachia infections occur within a broad range of invertebrates, these strategies are potentially applicable to a variety of medically and economically important insects. Here, we examine the interaction between Wolbachia infection frequency and host population size. We use a model to describe natural invasions of Wolbachia infections, artificial releases of infected hosts and releases of sterile males, as part of a traditional sterile insect technique programme. Model simulations demonstrate the importance of understanding the reproductive rate and intraspecific competition type of the targeted population, showing that releases of sterile or incompatible individuals may cause an undesired increase in the adult number. In addition, the model suggests a novel applied strategy that employs Wolbachia infections to suppress host populations. Releases of Wolbachia-infected hosts can be used to sustain artificially an unstable coexistence of multiple incompatible infections within a host population, allowing the host population size to be reduced, maintained at low levels, or eliminated.}, } @article {pmid11884755, year = {2002}, author = {Saint André, Av and Blackwell, NM and Hall, LR and Hoerauf, A and Brattig, NW and Volkmann, L and Taylor, MJ and Ford, L and Hise, AG and Lass, JH and Diaconu, E and Pearlman, E}, title = {The role of endosymbiotic Wolbachia bacteria in the pathogenesis of river blindness.}, journal = {Science (New York, N.Y.)}, volume = {295}, number = {5561}, pages = {1892-1895}, doi = {10.1126/science.1068732}, pmid = {11884755}, issn = {1095-9203}, support = {K08 AI054652/AI/NIAID NIH HHS/United States ; EY10320/EY/NEI NIH HHS/United States ; EY11373/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology/therapeutic use ; Brugia malayi/physiology ; Cornea/immunology/metabolism/microbiology/parasitology ; Dipetalonema/physiology ; Doxycycline/pharmacology/therapeutic use ; *Drosophila Proteins ; Eosinophils/immunology ; Humans ; Immunity, Innate ; Keratitis/immunology/microbiology/parasitology/pathology ; Membrane Glycoproteins/genetics/metabolism ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Microscopy, Confocal ; Neutrophil Infiltration ; Neutrophils/immunology ; Onchocerca volvulus/immunology/*microbiology/physiology ; Onchocerciasis, Ocular/*immunology/*microbiology/parasitology/pathology ; Receptors, Cell Surface/genetics/metabolism ; *Symbiosis ; Toll-Like Receptor 4 ; Toll-Like Receptors ; Wolbachia/immunology/*pathogenicity/physiology ; }, abstract = {Parasitic filarial nematodes infect more than 200 million individuals worldwide, causing debilitating inflammatory diseases such as river blindness and lymphatic filariasis. Using a murine model for river blindness in which soluble extracts of filarial nematodes were injected into the corneal stroma, we demonstrated that the predominant inflammatory response in the cornea was due to species of endosymbiotic Wolbachia bacteria. In addition, the inflammatory response induced by these bacteria was dependent on expression of functional Toll-like receptor 4 (TLR4) on host cells.}, } @article {pmid11884722, year = {2002}, author = {Pennisi, E}, title = {Infectious disease. New culprit emerges in river blindness.}, journal = {Science (New York, N.Y.)}, volume = {295}, number = {5561}, pages = {1809-1811}, doi = {10.1126/science.295.5561.1809}, pmid = {11884722}, issn = {1095-9203}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Blindness/etiology ; Eye/immunology/parasitology/*pathology ; Female ; Helminth Proteins/physiology ; Humans ; Keratitis ; Mice ; Microfilariae/immunology/physiology ; Onchocerca volvulus/growth & development/immunology/*microbiology/pathogenicity ; Onchocerciasis, Ocular/drug therapy/*immunology/*microbiology/parasitology ; Wolbachia/immunology/*pathogenicity/physiology ; }, } @article {pmid11880639, year = {2002}, author = {McGraw, EA and Merritt, DJ and Droller, JN and O'Neill, SL}, title = {Wolbachia density and virulence attenuation after transfer into a novel host.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {99}, number = {5}, pages = {2918-2923}, pmid = {11880639}, issn = {0027-8424}, support = {AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {*Adaptation, Physiological ; Animals ; Drosophila/*microbiology ; Drosophila melanogaster/*microbiology ; Female ; Male ; Virulence ; Wolbachia/growth & development/pathogenicity/*physiology ; }, abstract = {The factors that control replication rate of the intracellular bacterium Wolbachia pipientis in its insect hosts are unknown and difficult to explore, given the complex interaction of symbiont and host genotypes. Using a strain of Wolbachia that is known to over-replicate and shorten the lifespan of its Drosophila melanogaster host, we have tracked the evolution of replication control in both somatic and reproductive tissues in a novel host/Wolbachia association. After transinfection (the transfer of a Wolbachia strain into a different species) of the over-replicating Wolbachia popcorn strain from D. melanogaster to Drosophila simulans, we demonstrated that initial high densities in the ovaries were in excess of what was required for perfect maternal transmission, and were likely causing reductions in reproductive fitness. Both densities and fitness costs associated with ovary infection rapidly declined in the generations after transinfection. The early death effect in D. simulans attenuated only slightly and was comparable to that induced in D. melanogaster. This study reveals a strong host involvement in Wolbachia replication rates, the independence of density control responses in different tissues, and the strength of natural selection acting on reproductive fitness.}, } @article {pmid11861893, year = {2002}, author = {Lo, N and Casiraghi, M and Salati, E and Bazzocchi, C and Bandi, C}, title = {How many wolbachia supergroups exist?.}, journal = {Molecular biology and evolution}, volume = {19}, number = {3}, pages = {341-346}, doi = {10.1093/oxfordjournals.molbev.a004087}, pmid = {11861893}, issn = {0737-4038}, mesh = {Animals ; Arthropods/microbiology ; Biological Evolution ; Likelihood Functions ; Nematoda/microbiology ; *Phylogeny ; Polymerase Chain Reaction ; Symbiosis ; Wolbachia/*classification/*genetics ; }, } @article {pmid11856419, year = {2002}, author = {Kondo, N and Ijichi, N and Shimada, M and Fukatsu, T}, title = {Prevailing triple infection with Wolbachia in Callosobruchus chinensis (Coleoptera: Bruchidae).}, journal = {Molecular ecology}, volume = {11}, number = {2}, pages = {167-180}, doi = {10.1046/j.0962-1083.2001.01432.x}, pmid = {11856419}, issn = {0962-1083}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; Cloning, Molecular ; Coleoptera/*microbiology/physiology ; Female ; Japan ; Male ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Reproduction ; Sequence Analysis ; Wolbachia/classification/*genetics/*physiology ; }, abstract = {Prevailing triple infection with three distinct Wolbachia strains was identified in Japanese populations of the adzuki bean beetle, Callosobruchus chinensis. When a polymerase chain reaction (PCR) assay was conducted using universal primers for ftsZ and wsp, Wolbachia was detected in all the individuals examined, 288 males and 334 females from nine Japanese populations. PCR-restriction fragment length polymorphism (RFLP) analysis of cloned wsp gene fragments from single insects revealed that three types of wsp sequences coexist in the insects. Molecular phylogenetic analysis of the wsp sequences unequivocally demonstrated that C. chinensis harbours three phylogenetically distinct Wolbachia, tentatively designated as wBruCon, wBruOri and wBruAus, respectively. Diagnostic PCR analysis using specific primers demonstrated that, of 175 males and 235 females from nine local populations, infection frequencies with wBruCon, wBruOri and wBruAus were 100%, 96.3% and 97.0%, respectively. As for the infection status of individuals, triple infection (93.7%) dominated over double infection (6.1%) and single infection (0.2%). The amounts of wBruCon, wBruOri and wBruAus in field-collected adult insects were analysed by using a quantitative PCR technique in terms of wsp gene copies per individual insect. Irrespective of original populations, wBruCon and wBruOri (107 -108 wsp copies/insect) were consistently greater in amount than wBruAus (106 -107 wsp copies/insect), suggesting that the population sizes of the three Wolbachia strains are controlled, although the mechanism is unknown. Mating experiments suggested that the three Wolbachia cause cytoplasmic incompatibility at different levels of intensity.}, } @article {pmid11833290, year = {2001}, author = {Zakharov, IA and Shaikevich, EV}, title = {The Stockholm populations of Adalia bipunctata (L) (Coleoptera: Coccinellidae)--a case of extreme female-biased population sex ratio.}, journal = {Hereditas}, volume = {134}, number = {3}, pages = {263-266}, doi = {10.1111/j.1601-5223.2001.00263.x}, pmid = {11833290}, issn = {0018-0661}, mesh = {Animals ; Antigens, Bacterial/genetics ; Coleoptera/*microbiology/physiology ; DNA, Plant/analysis ; Female ; Male ; Pigmentation/genetics ; Pupa/microbiology ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/isolation & purification ; *Sex Ratio ; Spiroplasma/genetics/isolation & purification ; Sweden ; Symbiosis ; Wings, Animal ; Wolbachia/genetics/immunology ; }, abstract = {The genetic composition and sex ratio in the Stockholm populations of Adalia bipunctata have been studied. The overall frequency of melanics is 3.2%, which is significantly lower than in the populations of St. Petersburg and other large cities along the Baltic Sea. The secondary sex ratio in the Stockholm populations is female-biased 82:18. More than half of A. bipunctata females are infected with the male-killing Spiroplasma bacterium. Beetles of the co-existing species Adalia decempunctata are infected with a different bacterium belonging to the genus Rickettsia.}, } @article {pmid11823204, year = {2002}, author = {Dobson, SL and Marsland, EJ and Veneti, Z and Bourtzis, K and O'Neill, SL}, title = {Characterization of Wolbachia host cell range via the in vitro establishment of infections.}, journal = {Applied and environmental microbiology}, volume = {68}, number = {2}, pages = {656-660}, pmid = {11823204}, issn = {0099-2240}, support = {AI 40620/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Cell Culture Techniques/methods ; Cells, Cultured ; Culex/microbiology ; Drosophila/microbiology ; Drosophila melanogaster/*microbiology ; Lepidoptera/microbiology ; Spodoptera/*microbiology ; Wolbachia/*growth & development/isolation & purification/*pathogenicity ; }, abstract = {Maternally transmitted bacteria of the genus Wolbachia are obligate, intracellular symbionts that are frequently found in insects and cause a diverse array of reproductive manipulations, including cytoplasmic incompatibility, male killing, parthenogenesis, and feminization. Despite the existence of a broad range of scientific interest, many aspects of Wolbachia research have been limited to laboratories with insect-rearing facilities. The inability to culture these bacteria outside of the invertebrate host has also led to the existing bias of Wolbachia research toward infections that occur in host insects that are easily reared. Here, we demonstrate that Wolbachia infections can be simply established, stably maintained, and cryogenically stored in vitro using standard tissue culture techniques. We have examined Wolbachia host range by introducing different Wolbachia types into a single tissue culture. The results show that an Aedes albopictus (Diptera: Culicidae) cell line can support five different Wolbachia infection types derived from Drosophila simulans (Diptera: Drosophilidae), Culex pipiens (Culicidae), and Cadra cautella (Lepidoptera: Phycitidae). These bacterial types include infection types that have been assigned to two of the major Wolbachia clades. As an additional examination of Wolbachia host cell range, we demonstrated that a Wolbachia strain from D. simulans could be established in host insect cell lines derived from A. albopictus, Spodoptera frugiperda (Lepidoptera: Noctuidae), and Drosophila melanogaster. These results will facilitate the development of a Wolbachia stock center, permitting novel approaches for the study of Wolbachia infections and encouraging Wolbachia research in additional laboratories.}, } @article {pmid11812492, year = {2002}, author = {Ware, J and Moran, L and Foster, J and Posfai, J and Vincze, T and Guiliano, D and Blaxter, M and Eisen, J and Slatko, B}, title = {Sequencing and analysis of a 63 kb bacterial artificial chromosome insert from the Wolbachia endosymbiont of the human filarial parasite Brugia malayi.}, journal = {International journal for parasitology}, volume = {32}, number = {2}, pages = {159-166}, doi = {10.1016/s0020-7519(01)00367-8}, pmid = {11812492}, issn = {0020-7519}, mesh = {Animals ; Base Sequence ; Brugia malayi/*microbiology ; Chromosomes, Artificial, Bacterial/chemistry/*genetics ; DNA, Bacterial/chemistry/genetics ; Molecular Sequence Data ; Open Reading Frames/genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Wolbachia/chemistry/*genetics ; }, abstract = {Wolbachia endosymbiotic bacteria are widespread in filarial nematodes and are directly involved in the immune response of the host. In addition, antibiotics which disrupt Wolbachia interfere with filarial nematode development thus, Wolbachia provide an excellent target for control of filariasis. A 63.1 kb bacterial artificial chromosome insert, from the Wolbachia endosymbiont of the human filarial parasite Brugia malayi, has been sequenced using the New England Biolabs Inc. Genome Priming System() transposition kit in conjunction with primer walking methods. The bacterial artificial chromosome insert contains approximately 57 potential ORFs which have been compared by individual protein BLAST analysis with the 35 published complete microbial genomes in the Comprehensive Microbial Resource database at The Institute for Genomic Research and in the NCBI GenBank database, as well as to data from 22 incomplete genomes from the DOE Joint Genome Institute. Twenty five of the putative ORFs have significant similarity to genes from the alpha-proteobacteria Rickettsia prowazekii, the most closely related completed genome, as well as to the newly sequenced alpha-proteobacteria endosymbiont Sinorhizobium meliloti. The bacterial artificial chromosome insert sequence however has little conserved synteny with the R. prowazekii and S. meliloti genomes. Significant sequence similarity was also found in comparisons with the currently available sequence data from the Wolbachia endosymbiont of Drosophila melanogaster. Analysis of this bacterial artificial chromosome insert provides useful gene density and comparative genomic data that will contribute to whole genome sequencing of Wolbachia from the B. malayi host. This will also lead to a better understanding of the interactions between the endosymbiont and its host and will offer novel approaches and drug targets for elimination of filarial disease.}, } @article {pmid11811805, year = {2002}, author = {Sacchi, L and Corona, S and Casiraghi, M and Bandi, C}, title = {Does fertilization in the filarial nematode Dirofilaria immitis occur through endocytosis of spermatozoa?.}, journal = {Parasitology}, volume = {124}, number = {Pt 1}, pages = {87-95}, doi = {10.1017/s0031182001008964}, pmid = {11811805}, issn = {0031-1820}, mesh = {Animals ; Cell Membrane/physiology/ultrastructure ; Dirofilaria immitis/*physiology/ultrastructure ; Endocytosis/*physiology ; Female ; Fertilization/*physiology ; Male ; Microscopy, Electron ; Oocytes/physiology/ultrastructure ; Sperm-Ovum Interactions/physiology ; Spermatozoa/*physiology/ultrastructure ; }, abstract = {Information on the ultrastructural details of fertilization in filarial nematodes are still unavailable. Here we report new data on this process in Dirofilaria immits, the heartworm of dogs and cats. Electron microscopy allowed us to observe oocytes engulfing spermatozoa through an endocytosis-like process. We also observed spermatozoa inside the oocytes which still possessed their plasma membrane and which were clearly enveloped by a further membrane, likely derived from the endocytosis process. At this stage, at the interface between the sperm membrane and the endocytotic membrane (vacuolar space), we observed flocculent material in the proximity of the membranous organelles (MOs) of the sperm. In the proximity of the MOs, we also observed the enlargement of the vacuolar space. Other images showed the dissolution of the sperm membrane, and the release of nuclear masses and organelles in the egg cytoplasm. We did not observe the fusion of lysosomes to the endocytotic vacuoles. In addition, the lysis of the sperm organelles has never been observed inside the vacuoles containing the whole sperm. Thus we suggest that the degradation of the endocytotic and sperm plasma membranes is determined by material released by the MOs. Since we did not observe the entry of sperm into the oocytes by other mechanisms, we also suggest that endocytosis is the normal process used by the spermatozoon to get into the egg cytoplasm in D. immitis. Finally, during our observations of the seminal receptacle we did not observe any structure in the spermatozoa which could be interpreted as an intracellular bacterium. This is consistent with previous results indicating that the bacterium Wolbachia in filarial nematodes is not transmitted through the sperm.}, } @article {pmid11804774, year = {2002}, author = {Clark, ME and Veneti, Z and Bourtzis, K and Karr, TL}, title = {The distribution and proliferation of the intracellular bacteria Wolbachia during spermatogenesis in Drosophila.}, journal = {Mechanisms of development}, volume = {111}, number = {1-2}, pages = {3-15}, doi = {10.1016/s0925-4773(01)00594-9}, pmid = {11804774}, issn = {0925-4773}, mesh = {Animals ; Cysts/microbiology ; Drosophila/*microbiology ; Drosophila melanogaster ; Male ; Spermatogenesis ; Testis/*growth & development/*microbiology/pathology ; Wolbachia/*pathogenicity ; }, abstract = {Wolbachia is a cytoplasmically inherited alpha-proteobacterium found in a wide range of host arthropod and nematode taxa. Wolbachia infection in Drosophila is closely associated with the expression of a unique form of post-fertilization lethality termed cytoplasmic incompatibility (CI). This form of incompatibility is only expressed by infected males suggesting that Wolbachia exerts its effect during spermatogenesis. The growth and distribution of Wolbachia throughout sperm development in individual spermatocysts and elongating sperm bundles is described. Wolbachia growth within a developing cyst seems to begin during the pre-meiotic spermatocyte growth phase with the majority of bacteria accumulating during cyst elongation. Wolbachia are predominantly localized in the proximal end of the immature cyst, opposite the spermatid nuclei, and throughout development there appears little movement of Wolbachia between spermatids via the connecting cytoplasmic bridges. The overall number of new cysts infected as well as the number of spermatids/cysts infected seems to decrease with age and corresponds to the previously documented drop in CI with age. In contrast, in one CI expressing line of Drosophila melanogaster, fewer cysts are infected and a much greater degree of variation in numbers is observed between spermatids. Furthermore, the initiation and extent of the fastest period of Wolbachia growth in the D. melanogaster strain lags behind that of Drosophila simulans. The possible implications on the as yet unexplained mechanism of CI are discussed.}, } @article {pmid11779785, year = {2001}, author = {Charlat, S and Calmet, C and Merçot, H}, title = {On the mod resc model and the evolution of Wolbachia compatibility types.}, journal = {Genetics}, volume = {159}, number = {4}, pages = {1415-1422}, pmid = {11779785}, issn = {0016-6731}, mesh = {Algorithms ; Animals ; Arthropods ; Crosses, Genetic ; Cytoplasm/*microbiology/*physiology ; Evolution, Molecular ; Female ; Host-Parasite Interactions ; Male ; Models, Genetic ; Models, Theoretical ; Ovum/physiology ; Reproduction/physiology ; Spermatozoa/*microbiology ; Time Factors ; Wolbachia/*genetics/*physiology ; }, abstract = {Cytoplasmic incompatibility (CI) is induced by the endocellular bacterium Wolbachia. It results in an embryonic mortality occurring when infected males mate with uninfected females. The mechanism involved is currently unknown, but the mod resc model allows interpretation of all observations made so far. It postulates the existence of two bacterial functions: modification (mod) and rescue (resc). The mod function acts in the males' germline, before Wolbachia are shed from maturing sperm. If sperm is affected by mod, zygote development will fail unless resc is expressed in the egg. Interestingly, CI is also observed in crosses between infected males and infected females when the two partners bear different Wolbachia strains, demonstrating that mod and resc interact in a specific manner: Two Wolbachia strains are compatible with each other only if they harbor the same compatibility type. Here we focus on the evolutionary process involved in the emergence of new compatibility types from ancestral ones. We argue that new compatibility types are likely to evolve under a wider range of conditions than previously thought, through a two-step process. First, new mod variants can arise by mutation and spread by drift. This is possible because mod is expressed in males and Wolbachia is transmitted by females. Second, once such a mod variant achieves a certain frequency, it can create the conditions for the deterministic invasion of a new resc variant, allowing the invasion of a new mod resc pair. Furthermore, we show that a stable polymorphism might be maintained in natural populations, allowing the long-term existence of "suicidal" Wolbachia strains.}, } @article {pmid11761383, year = {2001}, author = {Dobson, SL and Rattanadechakul, W}, title = {A novel technique for removing Wolbachia infections from Aedes albopictus (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {38}, number = {6}, pages = {844-849}, doi = {10.1603/0022-2585-38.6.844}, pmid = {11761383}, issn = {0022-2585}, mesh = {Aedes/*microbiology ; Animals ; Anti-Bacterial Agents/*pharmacology ; Female ; Larva/microbiology ; Male ; Tetracycline/*pharmacology ; Wolbachia/*drug effects/genetics ; }, abstract = {Intracellular bacteria of the genus Wolbachia often behave as reproductive parasites by manipulating host reproduction to enhance the vertical transmission of infections. Wolbachia infections in Aedes albopictus (Skuse) cause a reproductive manipulation known as cytoplasmic incompatibility, which can reduce brood hatch. Because field populations of Ae. albopictus are naturally infected, studies of Wolbachia-induced effects on Ae. albopictus reproduction and fitness require that Wolbachia be artificially removed. Although simiple techniques for clearing Wolbachia infections from other host insects have been developed, removal of Wolbachia bacteria from Ae. abopictus is difficult. Here we describe an improved method for removing Wolbachia infections Ae. albopictus. This method differs from earlier techniques in that it relies upon the tetracycline treatment of adults instead of larvae. We demonstrate that tetracycline treatment of adult Ae. albopictus can predictably generate uninfected individuals, simplify the procedure required for Wolbachia removal, and reduce the level of inbreeding required to produce uninfected lines.}, } @article {pmid11760958, year = {2001}, author = {Dumler, JS and Barbet, AF and Bekker, CP and Dasch, GA and Palmer, GH and Ray, SC and Rikihisa, Y and Rurangirwa, FR}, title = {Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and 'HGE agent' as subjective synonyms of Ehrlichia phagocytophila.}, journal = {International journal of systematic and evolutionary microbiology}, volume = {51}, number = {Pt 6}, pages = {2145-2165}, doi = {10.1099/00207713-51-6-2145}, pmid = {11760958}, issn = {1466-5026}, mesh = {Anaplasma/classification/genetics ; Anaplasmataceae/*classification/genetics ; Animals ; Bacterial Proteins/*genetics ; Chaperonins/*genetics ; DNA, Ribosomal/genetics ; Ehrlichia/classification/genetics ; Ehrlichia ruminantium/classification/genetics ; Humans ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; Rickettsiaceae/*classification/genetics ; Sequence Alignment ; Sequence Analysis, DNA ; }, abstract = {The genera Anaplasma, Ehrlichia, Cowdria, Neorickettsia and Wolbachia encompass a group of obligate intracellular bacteria that reside in vacuoles of eukaryotic cells and were previously placed in taxa based upon morphological, ecological, epidemiological and clinical characteristics. Recent genetic analyses of 16S rRNA genes, groESL and surface protein genes have indicated that the existing taxa designations are flawed. All 16S rRNA gene and groESL sequences deposited in GenBank prior to 2000 and selected sequences deposited thereafter were aligned and phylogenetic trees and bootstrap values were calculated using the neighbour-joining method and compared with trees generated with maximum-probability, maximum-likelihood, majority-rule consensus and parsimony methods. Supported by bootstrap probabilities of at least 54%, 16S rRNA gene comparisons consistently clustered to yield four distinct clades characterized roughly as Anaplasma (including the Ehrlichia phagocytophila group, Ehrlichia platys and Ehrlichia bovis) with a minimum of 96.1% similarity, Ehrlichia (including Cowdria ruminantium) with a minimum of 97.7% similarity, Wolbachia with a minimum of 95.6% similarity and Neorickettsia (including Ehrlichia sennetsu and Ehrlichia risticii) with a minimum of 94.9% similarity. Maximum similarity between clades ranged from 87.1 to 94.9%. Insufficient differences existed among E. phagocytophila, Ehrlichia equi and the human granulocytic ehrlichiosis (HGE) agent to support separate species designations, and this group was at least 98.2% similar to any Anaplasma species. These 16S rRNA gene analyses are strongly supported by similar groESL clades, as well as biological and antigenic characteristics. It is proposed that all members of the tribes Ehrlichieae and Wolbachieae be transferred to the family Anaplasmataceae and that the tribe structure of the family Rickettsiaceae be eliminated. The genus Anaplasma should be emended to include Anaplasma (Ehrlichia) phagocytophila comb. nov. (which also encompasses the former E. equi and the HGE agent), Anaplasma (Ehrlichia) bovis comb. nov. and Anaplasma (Ehrlichia) platys comb. nov., the genus Ehrlichia should be emended to include Ehrlichia (Cowdria) ruminantium comb. nov. and the genus Neorickettsia should be emended to include Neorickettsia (Ehrlichia) risticii comb. nov. and Neorickettsia (Ehrlichia) sennetsu comb. nov.}, } @article {pmid11758285, year = {2001}, author = {Bazzocchi, C and Ceciliani, F and Brattig, N}, title = {[Bacterial symbionts (Wolbachia) of filarial nematodes: implications for the treatment and pathology of filariasis].}, journal = {Annali dell'Istituto superiore di sanita}, volume = {37}, number = {2}, pages = {265-273}, pmid = {11758285}, issn = {0021-2571}, mesh = {Animals ; Filariasis/*drug therapy/*etiology/immunology ; Filarioidea/immunology/*microbiology/pathogenicity ; Host-Parasite Interactions ; Humans ; Phylogeny ; *Rickettsiaceae/immunology/physiology ; }, abstract = {Filarial nematodes harbour intracellular, Gram-negative bacteria belonging to the genus Wolbachia. These bacteria have been observed in various species of filariae, including the main filariasis agents of humans and animals. It has been suggested that Wolbachia could play an important role in the biology of filarial nematodes and could be implicated in the pathogenesis of filarial diseases. Wolbachia could thus represent a target for the control of filariasis and key to the understanding of these diseases. Indeed, in various species of filariae, tetracycline treatments have been shown both to reduce/eliminate the Wolbachia population and to determine detrimental effects on the nematodes. In addition, proteins of Wolbachia have been shown to determine specific IgG responses in animals infected by filariae and some Wolbachia molecules (e.g. LPS) have been shown to stimulate innate-immunity responses (e.g. production of cytokines such as IL1, IL6, IL10, TNF-alpha and IFN-gamma by macrophages).}, } @article {pmid11749711, year = {2001}, author = {McGraw, EA and Merritt, DJ and Droller, JN and O'Neill, SL}, title = {Wolbachia-mediated sperm modification is dependent on the host genotype in Drosophila.}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1485}, pages = {2565-2570}, pmid = {11749711}, issn = {0962-8452}, support = {AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/microbiology/physiology ; Drosophila/genetics/*microbiology/physiology ; Drosophila melanogaster/genetics/microbiology/physiology ; Female ; Genotype ; Male ; Microinjections ; Spermatozoa/microbiology/*physiology ; Transfection ; Wolbachia/*pathogenicity ; }, abstract = {Estimates of Wolbachia density in the eggs, testes and whole flies of drosophilid hosts have been unable to predict the lack of cytoplasmic incompatibility (CI) expression in so-called mod(-) variants. Consequently, the working hypothesis has been that CI expression, although related to Wolbachia density, is also governed by unknown factors that are influenced by both host and bacterial genomes. Here, we compare the behaviour of the mod(-) over-replicating Wolbachia popcorn strain in its native Drosophila melanogaster host to the same strain transinfected into a novel host, namely Drosophila simulans. We report that (i) the popcorn strain is a close relative of other D. melanogaster infections, (ii) the mod(-) status of popcorn in D. melanogaster appears to result from its inability to colonize sperm bundles, (iii) popcorn is present in the bundles in D. simulans and induces strong CI expression, which demonstrates that the bacterial strain does not lack the genetic machinery for inducing CI and that there is host-species-specific control over Wolbachia tissue tropism, and (iv) infection of sperm bundles by the mod(-) D. simulans wCof strain indicates that there are several independent routes by which a strain can be a CI non-expressor.}, } @article {pmid11741630, year = {2001}, author = {Cross, HF and Haarbrink, M and Egerton, G and Yazdanbakhsh, M and Taylor, MJ}, title = {Severe reactions to filarial chemotherapy and release of Wolbachia endosymbionts into blood.}, journal = {Lancet (London, England)}, volume = {358}, number = {9296}, pages = {1873-1875}, doi = {10.1016/S0140-6736(01)06899-4}, pmid = {11741630}, issn = {0140-6736}, mesh = {Animals ; *Brugia malayi ; Diethylcarbamazine/*adverse effects ; Filariasis/blood/*drug therapy ; Filaricides/*adverse effects ; Humans ; Polymerase Chain Reaction ; Severity of Illness Index ; Systemic Inflammatory Response Syndrome/*chemically induced/classification/microbiology ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia bacteria seem to have evolved as essential endosymbionts of their filarial nematode hosts. Studies in mice have suggested that these bacteria are associated with systemic inflammatory reactions to filarial chemotherapy. We took blood samples from 15 Indonesian patients before and after treatment with diethylcarbamazine for Brugia malayi infection, and recorded the severity of any post-treatment inflammatory reactions. Blood from all three patients with severe adverse reactions and from one of six with moderate reactions was positive for Wolbachia DNA 4-48 h after diethylcarbamazine treatment. We suggest that these severe inflammatory reactions are associated with the release of endosymbionts into the blood after treatment for filariasis.}, } @article {pmid11737291, year = {2001}, author = {Morimoto, S and Nakai, M and Ono, A and Kunimi, Y}, title = {Late male-killing phenomenon found in a Japanese population of the oriental tea tortrix, Homona magnanima (Lepidoptera: Tortricidae).}, journal = {Heredity}, volume = {87}, number = {Pt 4}, pages = {435-440}, doi = {10.1046/j.1365-2540.2001.00924.x}, pmid = {11737291}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Female ; Gene Transfer, Horizontal ; Japan ; Larva/genetics/physiology ; Lepidoptera/*genetics/*physiology ; Male ; Oocytes/physiology ; Polymerase Chain Reaction ; Pupa/genetics/physiology ; *Sex Ratio ; }, abstract = {A female-biased sex ratio was found in the oriental tea tortrix, Homona magnanima (Lepidoptera: Tortricidae), in Tsukuba, Ibaraki, Japan. There was no difference in mean egg hatch between the all-female and normal strains. Greater than 50% mortality was observed in the all-female strain larvae, suggesting that female-only broods are produced as a result of late male-killing. The female-biased sex ratio was maternally inherited and maintained, even when females were backcrossed with males of the normal strain, thus implicating cytoplasmic parasitism as its cause. The phenomenon was persistent in the presence of antibiotics, and was not due to infection by agents that cause other male-killing phenomena, such as Rickettsia, Wolbachia, Spiroplasma, or protozoan parasites. When a homogenate of dead male larvae of the all-female strain was inoculated in normal-strain larvae, this male-killing trait was transmitted to the next generation; thus, its causative agent is probably transmitted horizontally as well.}, } @article {pmid11737290, year = {2001}, author = {Michel-Salzat, A and Cordaux, R and Bouchon, D}, title = {Wolbachia diversity in the Porcellionides pruinosus complex of species (Crustacea: Oniscidea): evidence for host-dependent patterns of infection.}, journal = {Heredity}, volume = {87}, number = {Pt 4}, pages = {428-434}, doi = {10.1046/j.1365-2540.2001.00920.x}, pmid = {11737290}, issn = {0018-067X}, mesh = {Animals ; Bacterial Infections/microbiology ; Base Sequence ; Biological Evolution ; Crustacea/*genetics/*microbiology ; DNA, Mitochondrial/genetics ; Ecosystem ; Genes, Bacterial/genetics ; Genetic Variation/*genetics ; Haplotypes/genetics ; Host-Parasite Interactions/*genetics ; Likelihood Functions ; Male ; Molecular Sequence Data ; Phylogeny ; Sequence Homology, Nucleic Acid ; Species Specificity ; Symbiosis/genetics ; Wolbachia/classification/*genetics ; }, abstract = {Porcellionides pruinosus is a cosmopolitan woodlouse. It is known to exhibit patterns of geographical variation between populations, and has been suspected to consist of several very closely related species. This species was found to carry Wolbachia endosymbionts, alpha-proteobacteria which are known to modify the reproduction of their crustacean hosts by inducing cytoplasmic incompatibility or feminization. In the P. pruinosus complex, Wolbachia induced feminization, but two different patterns of infection were reported: high prevalence and presence of infected males, or low prevalence and absence of infected males. In this study we investigated nine populations described as Porcellionides pruinosus carrying different Wolbachia strains, with the aim of investigating the possibility of coevolution between symbionts and hosts. Molecular analyses were carried out on both Wolbachia DNA (wsp gene) and host mitochondrial DNA (mt LSU rDNA). We show that (1) the nine host populations exhibited a high genetic differentiation so that they have to be split into two sibling species; (2) three different Wolbachia strains were found in the host complex but were not species-specific; and (3) the pattern of infection by Wolbachia was more host-dependent than symbiont-dependent.}, } @article {pmid11737276, year = {2001}, author = {Rokas, A and Atkinson, RJ and Brown, GS and West, SA and Stone, GN}, title = {Understanding patterns of genetic diversity in the oak gallwasp Biorhiza pallida: demographic history or a Wolbachia selective sweep?.}, journal = {Heredity}, volume = {87}, number = {Pt 3}, pages = {294-304}, doi = {10.1046/j.1365-2540.2001.00872.x}, pmid = {11737276}, issn = {0018-067X}, mesh = {Animals ; Base Sequence ; Cytochrome b Group/chemistry/genetics ; DNA Primers/chemistry ; DNA, Bacterial/analysis ; DNA, Mitochondrial/genetics ; Europe ; Female ; *Genetic Variation ; Haplotypes/genetics ; Isoenzymes/metabolism ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Wasps/*genetics/microbiology ; Wolbachia/*physiology ; }, abstract = {The endosymbiont Wolbachia can be responsible for selective sweeps on mitochondrial DNA variability within species. Similar signals can also result from demographic processes, although crucially the latter affect nuclear as well as mitochondrial loci. Here we present data on Wolbachia infection status and phylogeographic patterning for a widely distributed insect host, the oak gallwasp Biorhiza pallida (Hymenoptera: Cynipidae). Two hundred and eighteen females from eight European countries were screened for Wolbachia. All individuals from Hungary, Italy, France, U.K., Ireland, Switzerland, Sweden, and northern and southern Spain were infected with a single group A strain of Wolbachia, while populations in central Spain were not infected. A mitochondrial marker (cytochrome b) shows low variation and departure from neutrality in infected populations, but greater variation and no deviation from neutrality in Wolbachia-free populations. This pattern is compatible with a Wolbachia-induced selective sweep. However, we also find parallel differences between infected and uninfected populations for nuclear markers (sequence data for ITS1 and ITS2). All markers support the existence of a deep split between populations in Spain (some free of Wolbachia), and those in the rest of Europe (all infected). Allelic variation for five allozyme loci is also consistent with the Spain-rest of Europe split. Concordant patterns for nuclear and mitochondrial markers suggest that differences in the nature and extent of genetic diversity between these two regions are best explained by differing demographic histories (perhaps associated with range expansion from Pleistocene glacial refugia), rather than a Wolbachia-associated selective sweep.}, } @article {pmid11734058, year = {2001}, author = {Anderson, CL and Karr, TL}, title = {Wolbachia: evolutionary novelty in a rickettsial bacteria.}, journal = {BMC evolutionary biology}, volume = {1}, number = {}, pages = {10}, pmid = {11734058}, issn = {1471-2148}, mesh = {Chaperonin 60/chemistry/genetics ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Ehrlichia/genetics/pathogenicity ; *Evolution, Molecular ; Phylogeny ; Protein Structure, Secondary/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*genetics/*pathogenicity ; Symbiosis/physiology ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {BACKGROUND: Although closely related, the alpha-proteobacteria Wolbachia and the Rickettsiaceae (Rickettsia and Ehrlichia), employ different evolutionary life history strategies. Wolbachia are obligate endocellular symbionts that infect an extraordinary host range and, in contrast to the infectious and pathogenic Rickettsia and Ehrlichia, profoundly influence host reproductive biology.

RESULTS: Phylogenies of the Rickettsia, Ehrlichia, and Wolbachia were independently inferred from 16S rDNA sequences and GroEL amino acid sequences. Topologies inferred from both sets of sequence data were consistent with one another, and both indicate the genus Wolbachia shared a common ancestor most recently with Ehrlichia. These two genera are a sister group to the genus Rickettsia. Mapping biological properties onto this phylogeny reveals that manipulation of host reproduction, characteristic of Wolbachia strains, is a derived characteristic. This evolutionary novelty is accompanied by the loss of the ability to infect vertebrate hosts.

CONCLUSIONS: Because of the contrasting transmission strategies employed by each, Wolbachia is expected to maximize efficiency of vertical transmission, while Ehrlichia and Rickettsia will optimize horizontal transfer of infection. Wolbachia manipulation of host reproduction could thus be viewed as strategy employed by this bacterium to foster its own propagation via vertical transmission.}, } @article {pmid11703514, year = {2001}, author = {Keller, L and Liautard, C and Reuter, M and Brown, WD and Sundström, L and Chapuisat, M}, title = {Sex ratio and Wolbachia infection in the ant Formica exsecta.}, journal = {Heredity}, volume = {87}, number = {Pt 2}, pages = {227-233}, doi = {10.1046/j.1365-2540.2001.00918.x}, pmid = {11703514}, issn = {0018-067X}, mesh = {Animals ; Ants/*microbiology ; Female ; Male ; Polymerase Chain Reaction ; Sex Ratio ; Wolbachia/*physiology ; }, abstract = {Sex allocation data in social Hymenoptera provide some of the best tests of kin selection, parent-offspring conflict and sex ratio theories. However, these studies critically depend on controlling for confounding ecological factors and on identifying all parties that potentially manipulate colony sex ratio. It has been suggested that maternally inherited parasites may influence sex allocation in social Hymenoptera. If the parasites can influence sex allocation, infected colonies are predicted to invest more resources in females than non-infected colonies, because the parasites are transmitted through females but not males. Prime candidates for such sex ratio manipulation are Wolbachia, because these cytoplasmically transmitted bacteria have been shown to affect the sex ratio of host arthropods by cytoplasmic incompatibility, parthenogenesis, male-killing and feminization. In this study, we tested whether Wolbachia infection is associated with colony sex ratio in two populations of the ant Formica exsecta that have been the subject of extensive sex ratio studies. In these populations colonies specialize in the production of one sex or the other. We found that almost all F. exsecta colonies in both populations are infected with Wolbachia. However, in neither population did we find a significant association in the predicted direction between the prevalence of Wolbachia and colony sex ratio. In particular, colonies with a higher proportion of infected workers did not produce more females. Hence, we conclude that Wolbachia does not seem to alter the sex ratio of its hosts as a means to increase transmission rate in these two populations of ants.}, } @article {pmid11703513, year = {2001}, author = {Hurst, GD and Jiggins, FM and Robinson, SJ}, title = {What causes inefficient transmission of male-killing Wolbachia in Drosophila?.}, journal = {Heredity}, volume = {87}, number = {Pt 2}, pages = {220-226}, doi = {10.1046/j.1365-2540.2001.00917.x}, pmid = {11703513}, issn = {0018-067X}, mesh = {Animals ; Drosophila/genetics/*microbiology ; Environment ; Genetic Variation ; Sex Characteristics ; Sex Ratio ; Temperature ; Wolbachia/*physiology ; }, abstract = {Selfish genetic elements that distort the sex ratio are common in arthropods. Theory predicts they will invade and spread to fixation if they are vertically transmitted with perfect fidelity, potentially leading to host extinction. For inherited microorganisms that distort the sex ratio, inefficient vertical transmission or incomplete sex ratio distorting ability is required for host persistence. However, the relative roles of genetic and environmental factors in permitting the survival of male hosts and preventing parasite transmission are poorly understood. We examined the causes of transmission infidelity and male survival for a male-killing Wolbachia strain in Drosophila bifasciata. Under standard laboratory conditions (18 degrees C), in its standard genetic background, males are produced very rarely, and no case of reversion has been observed in 20 generations of laboratory rearing. To investigate the role of host genetic factors, Wolbachia was crossed into 27 different inbred lines of D. bifasciata, but in no case was reversion observed at preferred environmental temperatures. The role of elevated temperature in inducing inefficient transmission was examined. Whilst vertical transmission was perfect over three generations of maintenance at 23.5 degrees, transmission infidelity was observed at 25 degrees. We conclude that there is no evidence for the presence of either fixed or polymorphic host genes that repress transmission at standard environmental temperatures. However, severe temperature treatment does make vertical transmission imperfect. We suggest that the case of Wolbachia in D. bifasciata is one that is naturally balanced, the population being maintained polymorphic without the evolution of host resistance genes.}, } @article {pmid11685890, year = {2001}, author = {Taylor, M}, title = {One in the eye for river blindness.}, journal = {Trends in parasitology}, volume = {17}, number = {8}, pages = {358}, doi = {10.1016/s1471-4922(01)02063-3}, pmid = {11685890}, issn = {1471-4922}, mesh = {Animals ; Anthelmintics/therapeutic use ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Filaricides/*therapeutic use ; Humans ; Ivermectin/therapeutic use ; *Onchocerca volvulus ; Onchocerciasis, Ocular/*drug therapy ; Symbiosis/drug effects ; Wolbachia/*drug effects ; }, } @article {pmid11674872, year = {2001}, author = {Weeks, AR and Breeuwer, JA}, title = {Wolbachia-induced parthenogenesis in a genus of phytophagous mites.}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1482}, pages = {2245-2251}, pmid = {11674872}, issn = {0962-8452}, mesh = {Animals ; Bacterial Proteins/genetics ; *Cytoskeletal Proteins ; Female ; Male ; Mites/microbiology/*physiology ; Parthenogenesis/*physiology ; Reproduction ; Spermatozoa ; Tetracycline/pharmacology ; Wolbachia/genetics/*physiology ; }, abstract = {The vertically transmitted endosymbiotic bacterium Wolbachia modifies host reproduction in several ways in order to enhance its own spread. One such modification results in the induction of parthenogenesis, where males, which are unable to transmit Wolbachia, are not produced. Interestingly, parthenogenesis-inducing Wolbachia have only been found within haplodiploid insects and it is not known whether this exclusivity is the result of functional constraints of Wolbachia. Here we find a unique pattern of Wolbachia infection that is associated with parthenogenesis in six species within the phytophagous mite genus Bryobia. Through antibiotic treatment we show that, in two species, Bryobia praetiosa and an unidentified species, the Wolbachia infection is strictly associated with parthenogenesis. Microsatellite loci show the mechanism of parthenogenesis to be functionally apomictic and not gamete duplication, with progeny identical to their infected mother. Crossing experiments within B. praetiosa showed no evidence of sexual reproduction. These results are discussed with reference to the distribution of parthenogenesis-inducing Wolbachia and the diversification of the Bryobia genus.}, } @article {pmid11595053, year = {2001}, author = {Olsen, K and Reynolds, KT and Hoffmann, AA}, title = {A field cage test of the effects of the endosymbiont Wolbachia on Drosophila melanogaster.}, journal = {Heredity}, volume = {86}, number = {Pt 6}, pages = {731-737}, doi = {10.1046/j.1365-2540.2001.00892.x}, pmid = {11595053}, issn = {0018-067X}, mesh = {Analysis of Variance ; Animals ; Climate ; Drosophila melanogaster/*microbiology/*physiology ; Female ; Fertility ; Male ; Queensland ; Seasons ; Survival Rate ; *Symbiosis ; Victoria ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Wolbachia endosymbionts are known to affect the fitness of their hosts, but most of this information is from laboratory studies. In Drosophila melanogaster, Wolbachia frequencies vary clinically in frequency in Australia and may confound climatic adaptation. Here we use field cages in a reciprocal exchange design to test for Wolbachia effects in D. melanogaster in winter at temperate and tropical sites. Infected flies of both populations had a lower fecundity in tropical north Queensland, whilst in temperate southern Victoria Wolbachia effects depended on the nuclear population background. Here infected flies from Victoria were more fecund. Wolbachia also influenced larval/pupal viability in the tropics but this was dependent on population background. In comparisons of the populations, there was no evidence for local adaptation for total fecundity, viability or survival over winter. However, in Victoria, a local population had a higher late-life fecundity than a tropical population from Queensland that had higher early-life fecundity. At a tropical site, local Queensland flies had a higher early fecundity than Victorian flies. In contrast to many laboratory studies, mortality rates in the field cages increased only slightly over time. Both the Wolbachia effects and population differences have not been previously detected in laboratory studies with D. melanogaster and highlight the utility of Drosophila field studies in fitness experiments.}, } @article {pmid11592990, year = {2001}, author = {Zchori-Fein, E and Gottlieb, Y and Kelly, SE and Brown, JK and Wilson, JM and Karr, TL and Hunter, MS}, title = {A newly discovered bacterium associated with parthenogenesis and a change in host selection behavior in parasitoid wasps.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {98}, number = {22}, pages = {12555-12560}, pmid = {11592990}, issn = {0027-8424}, mesh = {Animals ; Base Sequence ; Female ; Microscopy, Electron ; Molecular Sequence Data ; Ovary/ultrastructure ; Oviposition ; Phylogeny ; Polymerase Chain Reaction ; Wasps/*microbiology ; Wolbachia/*classification/genetics/physiology ; }, abstract = {The symbiotic bacterium Wolbachia pipientis has been considered unique in its ability to cause multiple reproductive anomalies in its arthropod hosts. Here we report that an undescribed bacterium is vertically transmitted and associated with thelytokous parthenogenetic reproduction in Encarsia, a genus of parasitoid wasps. Although Wolbachia was found in only one of seven parthenogenetic Encarsia populations examined, the "Encarsia bacterium" (EB) was found in the other six. Among seven sexually reproducing populations screened, EB was present in one, and none harbored Wolbachia. Antibiotic treatment did not induce male production in Encarsia pergandiella but changed the oviposition behavior of females. Cured females accepted one host type at the same rate as control females but parasitized significantly fewer of the other host type. Phylogenetic analysis based on the 16S rDNA gene sequence places the EB in a unique clade within the Cytophaga-Flexibacter-Bacteroid group and shows EB is unrelated to the Proteobacteria, where Wolbachia and most other insect symbionts are found. These results imply evolution of the induction of parthenogenesis in a lineage other than Wolbachia. Importantly, these results also suggest that EB may modify the behavior of its wasp carrier in a way that enhances its transmission.}, } @article {pmid11580031, year = {2001}, author = {Vavre, F and Dedeine, F and Quillon, M and Fouillet, P and Fleury, F and Bouletreau, M}, title = {Within-species diversity of Wolbachia-induced cytoplasmic incompatibility in haplodiploid insects.}, journal = {Evolution; international journal of organic evolution}, volume = {55}, number = {8}, pages = {1710-1714}, doi = {10.1111/j.0014-3820.2001.tb00691.x}, pmid = {11580031}, issn = {0014-3820}, mesh = {Animals ; Cell Survival ; Crosses, Genetic ; Cytoplasm/physiology ; Drosophila melanogaster/growth & development/parasitology ; Embryo, Nonmammalian/physiology ; Female ; Male ; Ovum/physiology ; Sex Ratio ; Symbiosis ; Wasps/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia-induced cytoplasmic incompatibility (CI) can have two consequences in haplodiploid insects: fertilized eggs either die (female mortality, FM) or they develop into haploid males (male development, MD). Origin of this diversity remains poorly understood, but current hypotheses invoke variation in damage suffered by paternal chromosomes in incompatible eggs, thus intermediate CI types should be expected. Here, we show the existence of such a particular CI type. In the parasitoid wasp Leptopilina heterotoma, we compared CI effects in crosses involving lines derived from a single inbred line with various Wolbachia infection statuses (natural tri-infection, mono-infection, or no infection). Tri-infected males induce a FM CI type when crossed with either uninfected or mono-infected females. Crossing mono-infected males with uninfected females results in almost complete CI with both reduced offspring production, indicating partial mortality of fertilized eggs, and increased number of sons, showing haploid male development of others. Mono-infected males thus induce an intermediate Cl type when mated with uninfected females. The first evidence of this expected particular CI type demonstrates that no discontinuity separates MD and FM CI types, which appear to be end points of a phenotypic continuum. Second, different CI types can occur within a given species and even within offspring of a single pair. Third, phenotypic expression of the particular CI type induced by a given Wolbachia variant depends on other bacterial variants that co-infect the same tissues. These results support the idea that haplodiploids should be helpful in clarifying evolutionary pathways of insect-Wolbachia associations.}, } @article {pmid11560895, year = {2001}, author = {Rand, DM and Clark, AG and Kann, LM}, title = {Sexually antagonistic cytonuclear fitness interactions in Drosophila melanogaster.}, journal = {Genetics}, volume = {159}, number = {1}, pages = {173-187}, pmid = {11560895}, issn = {0016-6731}, mesh = {Alleles ; Analysis of Variance ; Animals ; Cell Nucleus/metabolism ; Cytoplasm/*metabolism ; DNA, Mitochondrial/metabolism ; Drosophila melanogaster/*chemistry ; Fathers ; Female ; Genetics, Population ; Genotype ; Haplotypes ; Male ; Models, Genetic ; Mothers ; Phenotype ; Protein Binding ; Sex Factors ; *Sexual Behavior, Animal ; *X Chromosome ; }, abstract = {Theoretical and empirical studies have shown that selection cannot maintain a joint nuclear-cytoplasmic polymorphism within a population except under restrictive conditions of frequency-dependent or sex-specific selection. These conclusions are based on fitness interactions between a diploid autosomal locus and a haploid cytoplasmic locus. We develop a model of joint transmission of X chromosomes and cytoplasms and through simulation show that nuclear-cytoplasmic polymorphisms can be maintained by selection on X-cytoplasm interactions. We test aspects of the model with a "diallel" experiment analyzing fitness interactions between pairwise combinations of X chromosomes and cytoplasms from wild strains of Drosophila melanogaster. Contrary to earlier autosomal studies, significant fitness interactions between X chromosomes and cytoplasms are detected among strains from within populations. The experiment further demonstrates significant sex-by-genotype interactions for mtDNA haplotype, cytoplasms, and X chromosomes. These interactions are sexually antagonistic--i.e., the "good" cytoplasms in females are "bad" in males--analogous to crossing reaction norms. The presence or absence of Wolbachia did not alter the significance of the fitness effects involving X chromosomes and cytoplasms but tended to reduce the significance of mtDNA fitness effects. The negative fitness correlations between the sexes demonstrated in our empirical study are consistent with the conditions that maintain cytoplasmic polymorphism in simulations. Our results suggest that fitness interactions with the sex chromosomes may account for some proportion of cytoplasmic variation in natural populations. Sexually antagonistic selection or reciprocally matched fitness effects of nuclear-cytoplasmic genotypes may be important components of cytonuclear fitness variation and have implications for mitochondrial disease phenotypes that differ between the sexes.}, } @article {pmid11555254, year = {2001}, author = {Noda, H and Miyoshi, T and Zhang, Q and Watanabe, K and Deng, K and Hoshizaki, S}, title = {Wolbachia infection shared among planthoppers (Homoptera: Delphacidae) and their endoparasite (Strepsiptera: Elenchidae): a probable case of interspecies transmission.}, journal = {Molecular ecology}, volume = {10}, number = {8}, pages = {2101-2106}, doi = {10.1046/j.0962-1083.2001.01334.x}, pmid = {11555254}, issn = {0962-1083}, mesh = {Animals ; Genes, Bacterial ; Insecta/*microbiology/*parasitology/physiology ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/*genetics/*physiology ; }, abstract = {Wolbachia, a group of parasitic bacteria of arthropods, are believed to be horizontally transmitted among arthropod taxa. We present a new probable example of interspecies horizontal transmission of Wolbachia by way of an endoparasite based on the conformity of Wolbachia gene sequences. Field samples of two rice planthoppers, Laodelphax striatellus and Sogatella furcifera possessed identical Wolbachia. Among three major endoparasites of planthoppers, a strepsipteran, Elenchus japonicus, harboured the identical Wolbachia strain, suggesting strepsipteran transmission of Wolbachia from one planthopper to the other. No Wolbachia was detected in a mermithid nematode Agamermis unka, and dryinid wasps possessed different types of Wolbachia.}, } @article {pmid11516587, year = {2001}, author = {Bandi, C and Trees, AJ and Brattig, NW}, title = {Wolbachia in filarial nematodes: evolutionary aspects and implications for the pathogenesis and treatment of filarial diseases.}, journal = {Veterinary parasitology}, volume = {98}, number = {1-3}, pages = {215-238}, doi = {10.1016/s0304-4017(01)00432-0}, pmid = {11516587}, issn = {0304-4017}, mesh = {Animals ; Anthelmintics/adverse effects/therapeutic use ; Anti-Bacterial Agents/therapeutic use ; Antibodies, Bacterial/analysis/biosynthesis ; Biological Evolution ; Diethylcarbamazine/adverse effects/therapeutic use ; Filariasis/drug therapy/*microbiology/parasitology ; Filarioidea/*microbiology ; Humans ; Phylogeny ; Symbiosis ; Wolbachia/drug effects/genetics/*growth & development ; }, abstract = {The presence of intracellular bacteria in the body of various species of filarial nematodes, including important parasites such as Brugia malayi, Dirofilaria immitis, and Onchocerca volvulus, was observed as early as the mid-1970s. These bacteria were shown to be transovarially transmitted (from the female worm to the offspring) and to be present in significant amounts in the body of the nematode. As highlighted by their discoverers, the potential importance of these bacteria is fairly obvious: (1) bacteria-derived molecules should be considered as having an immunological and pathological role in filarial diseases; (2) the interaction between the bacteria and the filarial host deserves investigation, in view of the possibility that the bacteria are needed by the host nematode and could thus represent a target for therapy. Other authors, independently from the discovery of these intracellular bacteria, showed that the antibiotic tetracycline (which is well known for its efficacy on intracellular bacteria) had detrimental effects on two species of filarial nematodes (Brugia pahangi and Litomosoides sigmodontis). It is therefore surprising that for more than 20 years, no further investigations focused on the bacteria of filarial nematodes, nor on the anti-filarial properties of tetracycline. Recently, the bacteria of filarial nematodes have been independently "rediscovered" by research groups from the schools of Hamburg, Liverpool and Milan. These bacteria are now classified as Wolbachia, and the basic aspects of their phylogenetic history and relationship with the Wolbachia of arthropods have been reconstructed. In addition, their implications for the pathogenesis and treatment of filarial diseases have started to be uncovered. This paper, which is authored by representatives of the three European schools who reopened this research area, reviews our present knowledge of these fascinating microorganisms, highlighting the complexity of a symbiotic system which involves, in addition to the nematode and its bacterium, the vertebrate host.}, } @article {pmid11488969, year = {2001}, author = {Moret, Y and Juchault, P and Rigaud, T}, title = {Wolbachia endosymbiont responsible for cytoplasmic incompatibility in a terrestrial crustacean: effects in natural and foreign hosts.}, journal = {Heredity}, volume = {86}, number = {Pt 3}, pages = {325-332}, doi = {10.1046/j.1365-2540.2001.00831.x}, pmid = {11488969}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Crustacea/*microbiology/*physiology ; Cytoplasm/physiology ; Embryonic Development ; Female ; Male ; Mitosis ; Reproduction ; Sexual Behavior, Animal ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are vertically transmitted endosymbionts that disturb the reproduction of many arthropods thereby enhancing their spread in host populations. Wolbachia are often responsible for changes of sex ratios in terrestrial isopods, a result of the feminization of genotypic males. Here we found that the Wolbachia hosted by Cylisticus convexus (wCc) caused unidirectional cytoplasmic incompatibility (CI), an effect commonly found in insects. To understand the diversity of Wolbachia-induced effects in isopods, wCc were experimentally transferred in a novel isopod host, Armadillidium vulgare. wCc conserved the ability to induce CI. However, Wolbachia were not transmitted to the eggs, so the capacity to restore the compatibility in crosses involving two transinfected individuals was lost. The feminizing Wolbachia hosted by A. vulgare was unable to rescue CI induced by wCc. These results showed that Wolbachia in isopods did not evolved broadly to induce feminization, and that CI and the feminizing effect are probably due to different mechanisms. In addition, wCc reduces the mating capacity of infected C. convexus males, suggesting that the bacteria might alter reproductive behaviour. The maintenance of wCc in host populations is discussed.}, } @article {pmid11485804, year = {2001}, author = {Charlat, S and Merçot, H}, title = {Cytoplasmic incompatibility and maternal-haploid.}, journal = {Trends in genetics : TIG}, volume = {17}, number = {8}, pages = {440-441}, doi = {10.1016/s0168-9525(01)02423-4}, pmid = {11485804}, issn = {0168-9525}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm/*metabolism ; Drosophila melanogaster ; Fathers ; Female ; *Haploidy ; Homozygote ; Male ; *Models, Genetic ; Mothers ; Sex Factors ; Wolbachia/*genetics ; }, } @article {pmid11485684, year = {2001}, author = {Brouqui, P and Fournier, PE and Raoult, D}, title = {Doxycycline and eradication of microfilaremia in patients with loiasis.}, journal = {Emerging infectious diseases}, volume = {7}, number = {3 Suppl}, pages = {604-605}, doi = {10.3201/eid0707.010747}, pmid = {11485684}, issn = {1080-6040}, mesh = {Adolescent ; Animals ; Anti-Bacterial Agents/*therapeutic use ; Doxycycline/*therapeutic use ; Humans ; Loa/microbiology ; Loiasis/*drug therapy/parasitology ; Male ; Middle Aged ; Parasitemia/drug therapy ; Polymerase Chain Reaction ; RNA, Ribosomal, 18S/genetics ; Wolbachia/isolation & purification ; }, } @article {pmid11472559, year = {2001}, author = {Taylor, MJ and Cross, HF and Ford, L and Makunde, WH and Prasad, GB and Bilo, K}, title = {Wolbachia bacteria in filarial immunity and disease.}, journal = {Parasite immunology}, volume = {23}, number = {7}, pages = {401-409}, doi = {10.1046/j.1365-3024.2001.00400.x}, pmid = {11472559}, issn = {0141-9838}, mesh = {Animals ; Antigens, Bacterial/immunology ; Brugia malayi/immunology/*microbiology ; Cytokines/metabolism ; Elephantiasis, Filarial/drug therapy/*immunology ; Filaricides/adverse effects/therapeutic use ; Humans ; Leukocytes/immunology ; Lipopolysaccharides/*immunology ; Symbiosis ; Wolbachia/*immunology/physiology ; }, abstract = {Lymphatic filarial nematodes are infected with endosymbiotic Wolbachia bacteria. Lipopolysaccharide from these bacteria is the major activator of innate inflammatory responses induced directly by the parasite. Here, we propose a mechanism by which Wolbachia initiates acute inflammatory responses associated with death of parasites, leading to acute filarial lymphangitis and adverse reactions to antifilarial chemotherapy. We also speculate that repeated exposure to acute inflammatory responses and the chronic release of bacteria, results in damage to infected lymphatics and desensitization of the innate immune system. These events will result in an increased susceptibility to opportunistic infections, which cause acute dermatolymphangitis associated with lymphoedema and elephantiasis. The recognition of the contribution of endosymbiotic bacteria to filarial disease could be exploited for clinical intervention by the targeting of bacteria with antibiotics in an attempt to reduce the development of filarial pathology.}, } @article {pmid11472553, year = {2001}, author = {Maizels, RM and Blaxter, ML and Scott, AL}, title = {Immunological genomics of Brugia malayi: filarial genes implicated in immune evasion and protective immunity.}, journal = {Parasite immunology}, volume = {23}, number = {7}, pages = {327-344}, doi = {10.1046/j.1365-3024.2001.00397.x}, pmid = {11472553}, issn = {0141-9838}, mesh = {Animals ; Antigens, Bacterial ; *Antigens, Helminth/chemistry/genetics/physiology ; Brugia malayi/*genetics/*immunology/pathogenicity/physiology ; *Genes, Helminth ; *Helminth Proteins/chemistry/genetics/physiology ; Wolbachia/immunology ; }, abstract = {Filarial nematodes are metazoan parasites with genome sizes of> 100 million base pairs, probably encoding 15 000-20 000 genes. Within this considerable gene complement, it seems likely that filariae have evolved a spectrum of immune evasion products which underpin their ability to live for many years within the human host. Moreover, no suitable vaccine currently exists for human filarial diseases, and few markers have yet been established for diagnostic use. In this review, we bring together biochemical and immunological data on prominent filarial proteins with the exciting new information provided by the Filarial Genome Project's expressed sequence tag (EST) database. In this discussion, we focus on those genes with the highest immunological profile, such as inhibitors of host enzymes, cytokine homologues and stage-specific surface proteins, as well as products associated with the mosquito-borne infective larva which offer the best opportunity for an anti-filarial vaccine. These gene products provide a fascinating glimpse of the molecular repertoire which helminth parasites have evolved to manipulate and evade the mammalian immune response.}, } @article {pmid11472552, year = {2001}, author = {Devaney, E and Yazdanbakhsh, M}, title = {Prospects and challenges in lymphatic filariasis.}, journal = {Parasite immunology}, volume = {23}, number = {7}, pages = {323-325}, doi = {10.1046/j.1365-3024.2001.00393.x}, pmid = {11472552}, issn = {0141-9838}, mesh = {Animals ; Brugia/genetics/immunology/microbiology/physiology ; Elephantiasis, Filarial/*immunology/parasitology ; Humans ; Lymphatic System/immunology/parasitology ; Wolbachia/physiology ; Wuchereria bancrofti/genetics/immunology/microbiology/physiology ; }, } @article {pmid11452274, year = {2001}, author = {Knight, J}, title = {Meet the Herod bug.}, journal = {Nature}, volume = {412}, number = {6842}, pages = {12-14}, doi = {10.1038/35083744}, pmid = {11452274}, issn = {0028-0836}, mesh = {Animals ; Biological Evolution ; Female ; Invertebrates/microbiology ; Male ; Sex Characteristics ; Wolbachia/*physiology ; }, } @article {pmid11446511, year = {2001}, author = {Heddi, A and Charles, H and Khatchadourian, C}, title = {Intracellular bacterial symbiosis in the genus Sitophilus: the 'biological individual' concept revisited.}, journal = {Research in microbiology}, volume = {152}, number = {5}, pages = {431-437}, doi = {10.1016/s0923-2508(01)01216-5}, pmid = {11446511}, issn = {0923-2508}, mesh = {Animals ; Biological Evolution ; Coleoptera/genetics/*microbiology/physiology ; DNA, Bacterial ; DNA, Mitochondrial ; Eukaryotic Cells/microbiology ; Gammaproteobacteria/*genetics ; Genome, Bacterial ; Reproduction ; *Symbiosis/genetics ; Wolbachia/*genetics ; }, abstract = {Eukaryotic cells, as genetic entities, most often involve several physically associated genomes that direct the metabolic cell equilibrium. In the coleopteran insects of the genus Sitophilus, in addition to the nucleus and the mitochondrial genomes, two other intracellular bacterial genomes belonging to the alpha and the gamma groups of Proteobacteria are also present. Coexisting with the eukaryotic host cell genomes, they intervene in the physiology and reproduction of the host. They are both transmitted vertically to the progeny and exhibit different levels of symbiont integration in insects. Their coexistence within a eukaryotic cell system illustrates the genetic complexity of animal tissue and questions the concept of the 'biological individual'.}, } @article {pmid11443570, year = {2001}, author = {Punkosdy, GA and Dennis, VA and Lasater, BL and Tzertzinis, G and Foster, JM and Lammie, PJ}, title = {Detection of serum IgG antibodies specific for Wolbachia surface protein in rhesus monkeys infected with Brugia malayi.}, journal = {The Journal of infectious diseases}, volume = {184}, number = {3}, pages = {385-389}, doi = {10.1086/322023}, pmid = {11443570}, issn = {0022-1899}, mesh = {Animals ; Antibodies, Bacterial/*blood ; Antibody Specificity ; Bacterial Proteins/genetics/*immunology ; Base Sequence ; Cloning, Molecular ; Disease Models, Animal ; Disease Progression ; Humans ; Immunoglobulin G/*blood ; Lymphedema/etiology/immunology ; Macaca mulatta ; Male ; Membrane Proteins/genetics/immunology ; Molecular Sequence Data ; Recombinant Fusion Proteins/immunology ; Rickettsia Infections/blood/complications/*immunology ; Time Factors ; *Wolbachia/immunology ; }, abstract = {The mechanism of lymphedema development in individuals with lymphatic filariasis is presently poorly understood. To investigate whether Wolbachia, symbiotic bacteria living within filarial nematodes, may be involved in disease progression, Wolbachia-specific immune responses were assayed in a group of Brugia malayi-infected rhesus monkeys. Serum IgG antibodies specific for a major Wolbachia surface protein (WSP) were detected in 2 of 12 infected monkeys. It is interesting that both of these monkeys developed lymphedema after becoming amicrofilaremic. WSP-specific antibody responses were temporally associated with increases in antifilarial IgG1 antibodies as well as lymphedema development. These findings suggest that Wolbachia may be important in understanding disease caused by filarial worms.}, } @article {pmid11437528, year = {2001}, author = {Rigaud, T and Pennings, PS and Juchault, P}, title = {Wolbachia bacteria effects after experimental interspecific transfers in terrestrial isopods.}, journal = {Journal of invertebrate pathology}, volume = {77}, number = {4}, pages = {251-257}, doi = {10.1006/jipa.2001.5026}, pmid = {11437528}, issn = {0022-2011}, mesh = {Animals ; Crustacea/*microbiology ; Female ; Male ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are intracellular parasites, vertically transmitted from mothers to offspring through the cytoplasm of the eggs. They manipulate the reproduction of their hosts to increase in frequency in host populations. In terrestrial isopods for example, Wolbachia are responsible for the full feminization of putative males, therefore increasing the proportion of females, the sex by which they are transmitted. Vertical transmission, however, is not the only means for Wolbachia propagation. Infectious (i.e., horizontal) transmission between different host species or taxa is required to explain the fact that the phylogeny of Wolbachia does not parallel that of their hosts. The aim of this study was to investigate, by experimental transinfections, whether Wolbachia strains could be successfully transferred to a different, previously uninfected isopod host. While Wolbachia survived in all the studied recipient species, vertical transmission was efficient only in cases where donor and recipient species were closely related. Even in this case, Wolbachia strains did not always keep their ability to entirely feminize their host, a deficiency that can be link to a low bacterial density in the host tissues. In addition, Wolbachia infection was associated with a decrease in host fertility, except when the bacterial strain came from the same host population as the recipient animals. This suggest that Wolbachia could be adapted to local host populations. It therefore seems that isopod Wolbachia are highly adapted to their host and can hardly infect another species of hosts. The successful infection of a given Wolbachia strain into a new isopod host species therefore probably requires a strong selection on bacterial variants.}, } @article {pmid11435084, year = {2001}, author = {Taylor, M}, title = {One in the eye for river blindness.}, journal = {Trends in microbiology}, volume = {9}, number = {7}, pages = {310}, doi = {10.1016/s0966-842x(01)02111-4}, pmid = {11435084}, issn = {0966-842X}, mesh = {Animals ; Anti-Bacterial Agents/therapeutic use ; Filaricides/*therapeutic use ; Humans ; Ivermectin/*therapeutic use ; Onchocerca volvulus/*microbiology ; Onchocerciasis/*drug therapy ; Wolbachia/drug effects ; }, } @article {pmid11431554, year = {2001}, author = {Otto, SP and Jarne, P}, title = {Evolution. Haploids--hapless or happening?.}, journal = {Science (New York, N.Y.)}, volume = {292}, number = {5526}, pages = {2441-2443}, doi = {10.1126/science.1062890}, pmid = {11431554}, issn = {0036-8075}, mesh = {Alleles ; Animals ; *Biological Evolution ; Diploidy ; Female ; Gene Expression ; Gram-Negative Bacteria/*physiology ; *Haploidy ; Male ; Mites/*genetics/*microbiology/physiology ; Mutation ; Ovum/microbiology ; Parthenogenesis ; Reproduction ; Selection, Genetic ; Symbiosis ; Wolbachia/physiology ; }, } @article {pmid11429144, year = {2001}, author = {Jiggins, FM and von Der Schulenburg, JH and Hurst, GD and Majerus, ME}, title = {Recombination confounds interpretations of Wolbachia evolution.}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1474}, pages = {1423-1427}, doi = {10.1098/rspb.2001.1656}, pmid = {11429144}, issn = {0962-8452}, mesh = {*Biological Evolution ; Genes, Bacterial ; Humans ; Recombination, Genetic ; *Wolbachia ; }, abstract = {Wolbachia are vertically transmitted bacteria known from arthropods and nematode worms, which are maintained in host populations because they either physiologically benefit infected individuals or parasitically manipulate their reproduction. The different manipulation phenotypes are scattered across the Wolbachia phylogeny, suggesting that there have been multiple evolutions of similar phenotypes. This conclusion relies on the assumption of an absence of recombination between bacterial strains, so that the gene used to reconstruct the phylogeny reflects the evolutionary history of the genes involved in the trait. We tested for recombination by reconstructing the phylogeny of two Wolbachia genes from seven B-subdivision strains. The two genes produced mutually incompatible topologies, indicating that these lineages are subject to genetic recombination. This means that many evolutionary patterns inferred from Wolbachia phylogenies must be re-evaluated. Furthermore, recombination may be an important feature both in the evolution of the manipulation phenotypes and avoidance of Müller's ratchet. Finally, we discuss the implications of recombination for attempts to genetically engineer Wolbachia for use in the control of crop pests and human pathogens.}, } @article {pmid11422512, year = {2001}, author = {Behura, SK and Sahu, SC and Mohan, M and Nair, S}, title = {Wolbachia in the Asian rice gall midge, Orseolia oryzae (Wood-Mason): correlation between host mitotypes and infection status.}, journal = {Insect molecular biology}, volume = {10}, number = {2}, pages = {163-171}, doi = {10.1046/j.1365-2583.2001.00251.x}, pmid = {11422512}, issn = {0962-1075}, mesh = {Animals ; Base Sequence ; DNA, Bacterial/classification ; DNA, Mitochondrial/classification ; Diptera/*microbiology ; Female ; Genes, Bacterial ; Male ; Molecular Sequence Data ; Oryza ; Polymorphism, Genetic ; RNA, Ribosomal ; Sequence Analysis, DNA ; Wolbachia/classification/*genetics ; }, abstract = {Using a PCR-based method, we detected Wolbachia in the Asian rice gall midge. Furthermore, results showed that all females across all biotypes are infected with Wolbachia. However, all male flies are not infected and show different infection frequency in different biotypes. We have also identified three mitotypes, in the rice gall midge, based on DraI restriction pattern of a portion of the 12S rRNA gene that was PCR amplified using primers specific to this gene. All the females and infected male flies had type 1 mtDNA while uninfected males showed only type 2 or 3 mtDNA. Inheritance patterns of mtDNA revealed the existence of a correlation between mtDNA type and Wolbachia infection in the Asian rice gall midge. Evidence for paternal inheritance of mtDNA in Wolbachia-free gall midge is also presented.}, } @article {pmid11403387, year = {2001}, author = {Casiraghi, M and Favia, G and Cancrini, G and Bartoloni, A and Bandi, C}, title = {Molecular identification of Wolbachia from the filarial nematode Mansonella ozzardi.}, journal = {Parasitology research}, volume = {87}, number = {5}, pages = {417-420}, doi = {10.1007/s004360000368}, pmid = {11403387}, issn = {0932-0113}, mesh = {Animals ; DNA, Bacterial/analysis ; DNA, Protozoan/analysis ; DNA, Ribosomal/genetics ; Mansonella/*microbiology ; Phylogeny ; Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; Wolbachia/classification/*genetics/isolation & purification ; }, abstract = {Mansonella ozzardi, a filarial parasite of humans in Latin America, has been shown to harbour intracellular bacteria not yet identified. Here we show that these bacteria, like those of other filarial nematodes, belong to the genus Wolbachia (alpha 2 Proteobacteria; Rickettsiales). Their unambiguous placement in the Wolbachia group was shown by 16S rDNA sequence analysis. However, the exact position of the Wolbachia from M. ozzardi relative to the other wolbachiae is not clear. Indeed, 16S rDNA sequence analysis places this bacterium at a deep branch in Wolbachia evolution. It is interesting that analysis of the 5S rDNA gene spacer of the nematode host also suggests that the genus Mansonella, together with the genus Loa, could represent a deep-branching lineage in filarial evolution.}, } @article {pmid11394345, year = {2001}, author = {Hoerauf, A and Fleischer, B and Walter, RD}, title = {Of filariasis, mice and men.}, journal = {Trends in parasitology}, volume = {17}, number = {1}, pages = {4-5}, doi = {10.1016/s1471-4922(00)01849-3}, pmid = {11394345}, issn = {1471-4922}, mesh = {Animals ; Filariasis/diagnosis/drug therapy/genetics/*immunology ; Filarioidea/immunology ; Humans ; Mice ; Onchocerca volvulus/immunology ; Wolbachia/genetics/immunology ; }, } @article {pmid11384164, year = {2001}, author = {Pfarr, KM and Qazi, S and Fuhrman, JA}, title = {Nitric oxide synthase in filariae: demonstration of nitric oxide production by embryos in Brugia malayi and Acanthocheilonema viteae.}, journal = {Experimental parasitology}, volume = {97}, number = {4}, pages = {205-214}, doi = {10.1006/expr.2001.4613}, pmid = {11384164}, issn = {0014-4894}, support = {AI 02642/AI/NIAID NIH HHS/United States ; AI 33395/AI/NIAID NIH HHS/United States ; NS30566/NS/NINDS NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/embryology/*enzymology ; Dipetalonema/embryology/*enzymology ; Female ; Gerbillinae ; Immunohistochemistry ; Male ; Nitric Oxide/*biosynthesis ; Nitric Oxide Synthase/isolation & purification/*metabolism ; }, abstract = {The radical gas nitric oxide (NO) is synthesized by nitric oxide synthase (NOS) from l-arginine and molecular oxygen. Nitric oxide is an important signaling molecule in invertebrate and vertebrate systems. Previously we have shown that NOS is localized to more tissues in Brugia malayi than has been reported in Ascaris suum. In this paper, we analyze the distribution of NOS in Acanthocheilonema viteae, a filarial nematode that differs from B. malayi in that A. viteae females release microfilariae without a sheath. A. viteae is also one of a few filarial parasites without the Wolbachia intracellular endosymbiont. By use of a specific antibody, NOS was demonstrated in extracts of A. viteae and Dirofilaria immitis. The localization pattern of NOS in A. viteae was similar to that seen in B. malayi, with the enzyme localized to the body wall muscles of both sexes, developing spermatozoa, intrauterine sperm, and early embryos. By use of DAF-2, a fluorescent indicator specific for nitric oxide, the embryos of B. malayi and A. viteae were demonstrated to produce NO ex utero. The near identical staining patterns seen in A. viteae and B. malayi argue that NO is not produced by Wolbachia, nor is it produced by the nematodes in response to the infection. Localization of NOS to the sperm of filarial nematodes suggests a role for NO during fertilization as has been described for sea urchin and ascidian fertilization. Demonstration of the activity of embryonic NOS supports our earlier hypothesis that NO is a signaling molecule during embryogenesis in filarial nematodes.}, } @article {pmid11380661, year = {2001}, author = {Jiggins, FM and Hurst, GD and Schulenburg, JH and Majerus, ME}, title = {Two male-killing Wolbachia strains coexist within a population of the butterfly Acraea encedon.}, journal = {Heredity}, volume = {86}, number = {Pt 2}, pages = {161-166}, doi = {10.1046/j.1365-2540.2001.00804.x}, pmid = {11380661}, issn = {0018-067X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Butterflies/drug effects/*microbiology ; Female ; Host-Parasite Interactions/drug effects/genetics ; Male ; Phylogeny ; Sex Factors ; Sex Ratio ; Tetracycline/pharmacology ; Wolbachia/*genetics/physiology ; }, abstract = {Inherited bacteria that kill male hosts early in their development are known from five insect orders. We ask to what extent the incidence of male-killers might be restricted by the rate at which new host-parasite interactions arise, by testing whether multiple male-killers have invaded a single host species. In Uganda, the butterflies Acraea encedon and A. encedana are both infected by the same strain of male-killing Wolbachia and there was no evidence of variation within the population. In Tanzanian A. encedon however, two phylogenetically distinct strains of male-killing Wolbachia were found within the same population. If this pattern of male-killer polymorphism is found to be general across infected species, it suggests that new male-killing infections arise frequently on an evolutionary time scale. Whether this polymorphism is stable, and what forces may be maintaining it, are unknown.}, } @article {pmid11377205, year = {2001}, author = {Brattig, NW and Büttner, DW and Hoerauf, A}, title = {Neutrophil accumulation around Onchocerca worms and chemotaxis of neutrophils are dependent on Wolbachia endobacteria.}, journal = {Microbes and infection}, volume = {3}, number = {6}, pages = {439-446}, doi = {10.1016/s1286-4579(01)01399-5}, pmid = {11377205}, issn = {1286-4579}, mesh = {Adult ; Animals ; Chemotactic Factors/metabolism ; Chemotaxis, Leukocyte ; Humans ; Interleukin-8 ; Neutrophils/enzymology/*immunology ; Onchocerca volvulus/cytology/*immunology/*microbiology ; Onchocerciasis/*immunology/pathology ; Symbiosis ; Tumor Necrosis Factor-alpha ; Wolbachia/drug effects/*immunology ; }, abstract = {Unlike in many other helminth infections, neutrophilic granulocytes are major cellular components in the hosts immune response against filarial worms. The pathways that drive the immune response involving neutrophils are unclear. This study shows that Wolbachia endobacteria (detectable by polyclonal antibodies against endobacterial heat shock protein 60 and catalase and by polymerase chain reaction being sensitive to doxycycline treatment) are direct and indirect sources of signals accounting for neutrophil accumulation around adult Onchocerca volvulus filariae. Worm nodules from untreated onchocerciasis patients displayed a strong neutrophil infiltrate adjacent to the live adult worms. In contrast, in patients treated with doxycycline to eliminate the endobacteria from O. volvulus and to render the worms sterile, the neutrophil accumulation around live adult filariae was drastically reduced. Neutrophils were absent in worm nodules from the deer filaria Onchocerca flexuosa, a species which does not contain endobacteria. Extracts of O. volvulus extirpated from untreated patients showed neutrophil chemotactic activity and in addition, induced strong TNF-alpha and IL-8 production in human monocytes, in contrast to filarial extracts obtained after doxycycline treatment. Thus, neutrophil chemotaxis and activation are induced directly by endobacterial products and also indirectly via chemokine induction by monocytes. These results show that the neutrophil response is a characteristic of endobacteria-containing filariae.}, } @article {pmid11375098, year = {2001}, author = {Jiggins, FM and Bentley, JK and Majerus, ME and Hurst, GD}, title = {How many species are infected with Wolbachia? Cryptic sex ratio distorters revealed to be common by intensive sampling.}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1472}, pages = {1123-1126}, doi = {10.1098/rspb.2001.1632}, pmid = {11375098}, issn = {0962-8452}, mesh = {Animals ; Butterflies/*microbiology ; Female ; Sex Ratio ; Species Specificity ; Wolbachia/isolation & purification/*physiology ; }, abstract = {Inherited bacterial symbionts from the genus Wolbachia have attracted much attention by virtue of their ability to manipulate the reproduction of their arthropod hosts. The potential importance of these bacteria has been underlined by surveys, which have estimated that 17% of insect species are infected. We examined whether these surveys have systematically underestimated the proportion of infected species through failing to detect the low-prevalence infections that are expected when Wolbachia distorts the sex ratio. We estimated the proportion of species infected with Wolbachia within Acraea butterflies by testing large collections of each species for infection. Seven out of 24 species of Acraea were infected with Wolbachia. Four of these were infected with Wolbachia at high prevalence, a figure compatible with previous broad-scale surveys, whilst three carried low-prevalence infections that would have had a very low likelihood of being detected by previous sampling methods. This led us to conclude that sex-ratio-distorting Wolbachia may be common in insects that have an ecology and/or genetics that permit the invasion of these parasites and that previous surveys may have seriously underestimated the proportion of species that are infected.}, } @article {pmid11375084, year = {2001}, author = {Arakaki, N and Miyoshi, T and Noda, H}, title = {Wolbachia-mediated parthenogenesis in the predatory thrips Franklinothrips vespiformis (Thysanoptera: Insecta).}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1471}, pages = {1011-1016}, doi = {10.1098/rspb.2001.1628}, pmid = {11375084}, issn = {0962-8452}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Female ; Genes, Bacterial ; Insecta/microbiology/*physiology ; Male ; *Parthenogenesis ; Phylogeny ; Polymerase Chain Reaction/methods ; Temperature ; Tetracycline/pharmacology ; Wolbachia/classification/genetics/isolation & purification/*physiology ; }, abstract = {Wolbachia are bacterial endosymbionts in arthropods and filarial nematodes. They cause thelytoky, which is a form of parthenogenesis in which females produce females without males, in hymenopteran insects. Infection of this parthenogenesis-inducing Wolbachia has been restricted to the order Hymenoptera, but was found in another insect order, Thysanoptera. A parthenogenetic colony of a predatory thrips Franklinothrips vespiformis (Aeolothripidae) possessed B-group Wolbachia. Male progeny were produced from this thrips by heat and tetracycline treatments. Males produced motile sperm, which were transferred to the female spermatheca by mating. However, the mating did not affect the sex ratios of the next generation, suggesting that the sperm do not fertilize the eggs.}, } @article {pmid11372962, year = {2001}, author = {Dobson, SL and Marsland, EJ and Rattanadechakul, W}, title = {Wolbachia-induced cytoplasmic incompatibility in single- and superinfected Aedes albopictus (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {38}, number = {3}, pages = {382-387}, doi = {10.1603/0022-2585-38.3.382}, pmid = {11372962}, issn = {0022-2585}, mesh = {Aedes/*microbiology/physiology ; Animals ; Cytoplasm ; Female ; Male ; Oviposition ; Wolbachia/*physiology ; }, abstract = {Maternally inherited bacteria of the genus Volbachia can cause cytoplasmic incompatibility resulting in the developmental arrest of early embryos. Previous studies have shown that both single- and superinfections of Wolbachia naturally occur in populations of Aedes albopictus (Skuse). Here, we report crossing experiments using three infection types occurring in Ae. albopictus: uninfected, single-infected, and superinfected individuals. Crosses were monitored over the lifetime of adults to detect possible effects of host age on cytoplasmic incompatibility levels and infection virulence. Both single- and superinfections induced high levels of cytoplasmic incompatibility throughout the lifetime of Ae. albopictus, demonstrating that both the single- and superinfections are well adapted for invasion of Ae. albopictus populations. Superinfected females were the longest lived and had the highest oviposition rates, whereas in males, uninfected individuals were the longest lived. These latter results demonstrate the need for additional experiments to better elucidate Wolbachia effects on host fitness in addition to cytoplasmic incompatibility.}, } @article {pmid11356444, year = {2001}, author = {Hoerauf, A and Mand, S and Adjei, O and Fleischer, B and Büttner, DW}, title = {Depletion of wolbachia endobacteria in Onchocerca volvulus by doxycycline and microfilaridermia after ivermectin treatment.}, journal = {Lancet (London, England)}, volume = {357}, number = {9266}, pages = {1415-1416}, doi = {10.1016/S0140-6736(00)04581-5}, pmid = {11356444}, issn = {0140-6736}, mesh = {Animals ; Anti-Bacterial Agents/administration & dosage/*therapeutic use ; Case-Control Studies ; Doxycycline/administration & dosage/*therapeutic use ; Drug Therapy, Combination ; Female ; Filaricides/administration & dosage/*therapeutic use ; Humans ; Ivermectin/administration & dosage/*therapeutic use ; Onchocerca volvulus/*drug effects/microbiology ; Onchocerciasis/*drug therapy ; Skin/pathology ; Wolbachia/*drug effects ; }, abstract = {Ivermectin is the drug used for mass chemotherapy of onchocerciasis within the WHO African Programme for Onchocerciasis Control. This approach aims to eliminate the disease as a public health problem but using one dose per year may not completely interrupt transmission since it does not suppress microfilaridermia thoroughly enough. Here we show that additional treatment with doxycycline, previously shown to sterilise adult female worms for a few months by depletion of symbiotic wolbachia endobacteria, significantly enhances ivermectin-induced suppression of microfilaridermia, rendering anti-wolbachia treatment a promising basis for blocking transmission by a drug-based approach.}, } @article {pmid11355885, year = {2001}, author = {Masui, S and Kuroiwa, H and Sasaki, T and Inui, M and Kuroiwa, T and Ishikawa, H}, title = {Bacteriophage WO and virus-like particles in Wolbachia, an endosymbiont of arthropods.}, journal = {Biochemical and biophysical research communications}, volume = {283}, number = {5}, pages = {1099-1104}, doi = {10.1006/bbrc.2001.4906}, pmid = {11355885}, issn = {0006-291X}, mesh = {Animals ; Bacteriophages/genetics/*isolation & purification/ultrastructure ; Base Sequence ; Female ; Genome, Bacterial ; Gryllidae/*microbiology ; Lysogeny ; Molecular Sequence Data ; Open Reading Frames ; Ovary/microbiology ; Symbiosis/*physiology ; Wolbachia/genetics/*physiology/*virology ; }, abstract = {Wolbachia are intracellular symbionts mainly found in arthropods, causing various sexual alterations on their hosts by unknown mechanisms. Here we report the results that strongly suggest that Wolbachia have virus-like particles of phage WO, which was previously identified as a prophage-like element in the Wolbachia genome. Wolbachia (strain wTai) infection in an insect was detected with the antibody against Wsp, an outer surface protein of Wolbachia, by fluorescence microscopy and immunoelectron-microscopy for the first time. Virus-like particles in Wolbachia were observed by electron-microscopy. The 0.22-microm filtrate of insect ovary contained DAPI-positive particles, and PCR analysis demonstrated that a phage WO DNA passed through the filter while Wolbachia DNA were eliminated, suggesting that the DAPI-positive particles were phage WO.}, } @article {pmid11353833, year = {2001}, author = {Dedeine, F and Vavre, F and Fleury, F and Loppin, B and Hochberg, ME and Bouletreau, M}, title = {Removing symbiotic Wolbachia bacteria specifically inhibits oogenesis in a parasitic wasp.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {98}, number = {11}, pages = {6247-6252}, pmid = {11353833}, issn = {0027-8424}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Dose-Response Relationship, Drug ; Female ; Lipopolysaccharides ; Male ; Oocytes ; Oogenesis/drug effects/*physiology ; Rifampin/pharmacology ; Symbiosis ; Wasps/drug effects/microbiology/*physiology ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are bacteria that live in the cells of various invertebrate species to which they cause a wide range of effects on physiology and reproduction. We investigated the effect of Wolbachia infection in the parasitic wasp, Asobara tabida Nees (Hymenoptera, Braconidae). In the 13 populations tested, all individuals proved to be infected by Wolbachia. The removal of Wolbachia by antibiotic treatment had a totally unexpected effect-aposymbiotic female wasps were completely incapable of producing mature oocytes and therefore could not reproduce. In contrast, oogenesis was not affected in treated Asobara citri, a closely related species that does not harbor Wolbachia. No difference between natural symbiotic and cured individuals was found for other adult traits including male fertility, locomotor activity, and size, indicating that the effect on oogenesis is highly specific. We argue that indirect effects of the treatments used in our study (antibiotic toxicity or production of toxic agents) are very unlikely to explain the sterility of females, and we present results showing a direct relationship between oocyte production and Wolbachia density in females. We conclude that Wolbachia is necessary for oogenesis in these A. tabida strains, and this association would seem to be the first example of a transition from facultative to obligatory symbiosis in arthropod-Wolbachia associations.}, } @article {pmid11352061, year = {2001}, author = {Zimmer, C}, title = {Wolbachia. A tale of sex and survival.}, journal = {Science (New York, N.Y.)}, volume = {292}, number = {5519}, pages = {1093-1095}, doi = {10.1126/science.292.5519.1093}, pmid = {11352061}, issn = {0036-8075}, mesh = {Animals ; Butterflies/microbiology/physiology ; Culicidae/parasitology/physiology ; Disease Vectors ; Drosophila melanogaster/parasitology/physiology ; Ecosystem ; Female ; Genome, Bacterial ; Humans ; Infertility/microbiology/physiopathology ; Invertebrates/*microbiology/*physiology ; Malaria/parasitology/prevention & control ; Male ; Nematoda/microbiology/physiology ; Ovum/growth & development/microbiology ; Parasitic Diseases/*parasitology/prevention & control/transmission ; Reproduction ; Selection, Genetic ; Sex Distribution ; Species Specificity ; Symbiosis ; Trypanosomiasis, African/parasitology ; Virulence/genetics ; Wasps/microbiology/physiology ; Wolbachia/genetics/pathogenicity/*physiology ; }, } @article {pmid11345332, year = {2001}, author = {Fujii, Y and Kageyama, D and Hoshizaki, S and Ishikawa, H and Sasaki, T}, title = {Transfection of Wolbachia in Lepidoptera: the feminizer of the adzuki bean borer Ostrinia scapulalis causes male killing in the Mediterranean flour moth Ephestia kuehniella.}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1469}, pages = {855-859}, doi = {10.1098/rspb.2001.1593}, pmid = {11345332}, issn = {0962-8452}, mesh = {Androgen-Insensitivity Syndrome/microbiology ; Animals ; Base Sequence ; DNA Primers/genetics ; Female ; Lepidoptera/*microbiology ; Male ; Microinjections ; Phenotype ; Sex Ratio ; Transfection ; Wolbachia/*genetics/*pathogenicity ; }, abstract = {Two species of Lepidoptera, Ostrinia scapulalis and Ephestia kuehniella, harbour Wolbachia, which are maternally transmitted intracellular bacteria that often cause reproductive abnormalities in arthropods. While the infection in O. scapulalis causes conversion of genetic males into functional females (feminization), that in E. kuehniella induces cytoplasmic incompatibility. In the present study, we investigated the relative importance of host and Wolbachia factors in the differential expression of reproductive alterations in these insects. We transferred the Wolbachia harboured by O. scapulalis to E. kuehniella in which the original infection had been cured by tetracycline treatment. The transfected strain of E. kuehniella expressed a maternally inherited, female-biased sex ratio. Unexpectedly, two lines of evidence suggested that the sex ratio distortion was due to male killing. First, higher mortality of young larvae was observed. Second, the removal of the transferred Wolbachia resulted in the recovery of a 1:1 sex ratio, whereas the removal of a feminizer should result in a male-biased sex ratio among offspring. To the authors' knowledge, this is the first report that a single Wolbachia strain can cause two distinct sexual abnormalities in different hosts. Our observations highlighted the importance of host-Wolbachia interactions in determining the phenotype of reproductive alterations.}, } @article {pmid11328780, year = {2001}, author = {Hermans, PG and Hart, CA and Trees, AJ}, title = {In vitro activity of antimicrobial agents against the endosymbiont Wolbachia pipientis.}, journal = {The Journal of antimicrobial chemotherapy}, volume = {47}, number = {5}, pages = {659-663}, doi = {10.1093/jac/47.5.659}, pmid = {11328780}, issn = {0305-7453}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Cells, Cultured/microbiology ; Doxycycline/*pharmacology ; Humans ; Insecta ; Microbial Sensitivity Tests ; Rickettsiaceae Infections/microbiology ; Wolbachia/*drug effects ; }, abstract = {Arthropod-transmitted (filarial) nematodes are important causes of disease in humans in tropical countries, yet no safe drug appropriate for mass delivery kills the adult worms. However, most filarial nematodes contain rickettsia-like bacteria of the genus Wolbachia, and related bacteria also occur in insects. There is increasing evidence that these bacteria have significant functions in the biology of filarial nematodes. They are thus important targets in the search for antifilarial drugs and experiments in animals and humans have suggested that antibiotic therapy has potential in treating filarial infections. To optimize future clinical trials there is a need for a fast and simple in vitro drug screen to compare drug efficacies against Wolbachia. In the absence of Wolbachia-infected nematode cell lines, we have utilized an Aedes albopictus insect cell line, naturally infected with Wolbachia, to test the activity of antimicrobial agents. Of the five antibiotics tested, doxycycline, oxytetracycline and rifampicin showed good activity (MICs of 0.0625, 4 and 0.0625 mg/L, respectively) whereas ciprofloxacin and penicillin were shown to have no effect.}, } @article {pmid11301253, year = {2001}, author = {Werren, JH and Bartos, JD}, title = {Recombination in Wolbachia.}, journal = {Current biology : CB}, volume = {11}, number = {6}, pages = {431-435}, doi = {10.1016/s0960-9822(01)00101-4}, pmid = {11301253}, issn = {0960-9822}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; DNA, Bacterial/analysis ; Phylogeny ; *Recombination, Genetic ; Sequence Analysis, DNA ; Wasps/microbiology ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia are widely distributed intracellular bacteria that cause a number of reproductive alterations in their eukaryotic hosts. Such alterations include the induction of parthenogenesis, feminization, cytoplasmic incompatibility, and male killing [1-11]. These important bacteria may play a role in rapid speciation in insects [12-14], and there is growing interest in their potential uses as tools for biological control and genetic manipulation of pests and disease vectors [15-16]. Here, we show recombination in the Wolbachia outer surface protein gene (wsp) between strains of Wolbachia. In addition, we find a possible ecological context for this recombination. Evidence indicates either genetic exchange between Wolbachia in a parasitoid wasp and in the fly that it parasitizes or horizontal transfer of Wolbachia between the parasitoid and the fly, followed by a recombination event. Results have important implications for the evolution of these bacteria and the potential use of Wolbachia in biological control.}, } @article {pmid11298981, year = {2001}, author = {Mandel, MJ and Ross, CL and Harrison, RG}, title = {Do Wolbachia infections play a role in unidirectional incompatibilities in a field cricket hybrid zone?.}, journal = {Molecular ecology}, volume = {10}, number = {3}, pages = {703-709}, doi = {10.1046/j.1365-294x.2001.01213.x}, pmid = {11298981}, issn = {0962-1083}, mesh = {Animals ; Bacterial Proteins/chemistry/genetics ; Chimera/*genetics/physiology ; *Cytoskeletal Proteins ; DNA, Bacterial/chemistry/isolation & purification ; Female ; Gryllidae/*genetics/*microbiology ; Male ; New England ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Reproduction/genetics/physiology ; Sequence Alignment ; Sequence Analysis, DNA ; Wolbachia/chemistry/genetics/*physiology ; }, abstract = {Two closely related field crickets, Gryllus firmus and G. pennsylvanicus, hybridize along an extensive north-south zone in the eastern United States. Crosses between G. firmus males and G. pennsylvanicus females produce viable and fertile F1, but the reciprocal cross consistently fails to produce offspring. Wolbachia, a bacterial parasite of arthropods that causes unidirectional incompatibilities in a variety of insect species, has been suggested as the cause of the observed incompatibility between G. pennsylvanicus and G. firmus. We examine the presence/absence of Wolbachia strains, defined by sequencing the ftsZ gene, in four cricket populations from the north-eastern United States. Most G. firmus individuals are infected (100% in Guilford, Connecticut; 65% in Seaside Park, New Jersey) and > 95% of those infected harbour a single strain of Wolbachia. All individuals in G. pennsylvanicus populations (Ithaca, New York; Sharon, Connecticut) are infected; the majority of individuals carry a second strain of Wolbachia, but a significant fraction carry the same strain found commonly in G. firmus. The presence of an apparently identical Wolbachia strain in crickets of both species means that some crosses between G. pennsylvanicus males and G. firmus females should be compatible. We have no evidence of such compatibility. Furthermore, if Wolbachia infections are responsible for the observed incompatibility between species, then incompatibilities must also exist within G. pennsylvanicus, because this species harbours both Wolbachia strains. Although some single pair crosses within G. pennsylvanicus do fail to produce offspring, the proportion is lower than expected if Wolbachia were responsible. Therefore, Wolbachia is unlikely to be involved in reproductive isolation between the two cricket species.}, } @article {pmid11297179, year = {2001}, author = {Stouthamer, R and van Tilborg, M and de Jong, JH and Nunney, L and Luck, RF}, title = {Selfish element maintains sex in natural populations of a parasitoid wasp.}, journal = {Proceedings. Biological sciences}, volume = {268}, number = {1467}, pages = {617-622}, doi = {10.1098/rspb.2000.1404}, pmid = {11297179}, issn = {0962-8452}, mesh = {Animals ; Chromosome Mapping ; Crosses, Genetic ; Female ; Fertilization ; Karyotyping ; Larva ; Lepidoptera/*parasitology ; Male ; Ovum/physiology ; Parthenogenesis ; Population Dynamics ; Reproduction ; Sex Ratio ; Wasps/genetics/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {Genomic conflicts between heritable elements with different modes of inheritance are important in the maintenance of sex and in the evolution of sex ratio. Generally, we expect sexual populations to exhibit a 1:1 sex ratio. However, because of their biology, parasitoid wasps often exhibit a female-biased sex ratio. Sex-ratio distorters can further alter this optimum, sometimes leading to the complete loss of sexual reproduction. In the parasitoid wasp Trichogramma kaykai ca. 4-26% of females in field populations are infected with a bacterial sex-ratio distorter, Wolbachia, allowing virgin mothers to produce daughters. In some micro-Hymenoptera these infections have led to the complete loss of sex, but in field populations of T. kaykai the proportion of individuals infected remains relatively stable. We tested several hypotheses to explain this low infection level, including inefficient and horizontal transmission of Wolbachia, suppressor genes negating the effect of Wolbachia and the presence of male-biasing sex-ratio distorters. Here, a male-biasing sex-ratio distorter, a parasitic B chromosome, causing females to produce only sons, keeps the frequency of Wolbachia low. The male-biasing factor of T. kaykai is the second known case of a B chromosome manipulating the reproduction of a parasitoid wasp.}, } @article {pmid11296829, year = {2001}, author = {Ono, M and Braig, HR and Munstermann, LE and Ferro, C and O'Neill, SL}, title = {Wolbachia infections of phlebotomine sand flies (Diptera: Psychodidae).}, journal = {Journal of medical entomology}, volume = {38}, number = {2}, pages = {237-241}, doi = {10.1603/0022-2585-38.2.237}, pmid = {11296829}, issn = {0022-2585}, support = {AI 34521/AI/NIAID NIH HHS/United States ; AI 40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Base Sequence ; DNA, Bacterial ; Molecular Sequence Data ; Polymerase Chain Reaction/methods ; Psychodidae/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Old and New World phlebotomine sand fly species were screened for infection with Wolbachia, intracellular bacterial endosymbionts found in many arthropods and filarial nematodes. Of 53 samples representing 15 species, nine samples offour species were found positive for Wolbachia by polymerase chain reaction amplification using primers for the Wolbachia surface protein (wsp) gene. Five of the wsp gene fragments from four species were cloned, sequenced, and used for phylogenetic analysis. These wsp sequences were placed in three different clades within the arthropod associated Wolbachia (groups A and B), suggesting that Wolbacia has infected sand flies on more than one occasion. Two distantly related sand fly species, Lutzomyia (Psanthyromyia) shannoi (Dyar) and Lutzomyia (Nyssomyia) whitmani (Antunes & Coutinho), infected with an identical Wolbachia strain suggest a very recent horizontal transmission.}, } @article {pmid11267910, year = {2001}, author = {Noda, H and Koizumi, Y and Zhang, Q and Deng, K}, title = {Infection density of Wolbachia and incompatibility level in two planthopper species, Laodelphax striatellus and Sogatella furcifera.}, journal = {Insect biochemistry and molecular biology}, volume = {31}, number = {6-7}, pages = {727-737}, doi = {10.1016/s0965-1748(00)00180-6}, pmid = {11267910}, issn = {0965-1748}, mesh = {Animals ; DNA, Bacterial/analysis ; Genes, Bacterial ; Hemiptera/*microbiology ; Ovum/growth & development ; Polymerase Chain Reaction/methods ; Sequence Analysis, DNA ; Tetracycline ; Wolbachia/genetics/*isolation & purification/physiology ; }, abstract = {Wolbachia, a bacterial endosymbiote of arthropods, causes cytoplasmic incompatibility (CI) in many insect species. CI traits were studied in two planthopper species, Laodelphax striatellus and Sogatella furcifera, and Wolbachia densities in these planthopper species were calculated by quantitative PCR methods. The CI level of L. striatellus was quite high and even aged males strongly caused CI. In contrast, S. furcifera showed partial CI, and males lost their ability to cause CI with age. Wolbachia infecting these two planthopper species were the same with respect to the nucleotide sequences of Wolbachia genes, 16S rDNA, ftsZ gene, groE genes, and wsp gene. Two methods for quantitative PCR, one using a DNA sequencer and the other a real-time sequence detection system, were established to calculate the amount of Wolbachia in the planthoppers. The density of Wolbachia in S. furcifera males was quite low. The difference in CI levels between the two planthopper species seems to be due to different amounts of Wolbachia infecting males.}, } @article {pmid11253199, year = {2001}, author = {Smith, O}, title = {Evolution. Nota bene. Wolbachia and wasp evolution.}, journal = {Science (New York, N.Y.)}, volume = {291}, number = {5509}, pages = {1719}, doi = {10.1126/science.291.5509.1719}, pmid = {11253199}, issn = {0036-8075}, mesh = {Animals ; *Biological Evolution ; Female ; Fertility ; Male ; Reproduction ; Species Specificity ; *Symbiosis ; Wasps/*microbiology/*physiology ; Wolbachia/*physiology ; }, } @article {pmid11244060, year = {2001}, author = {Sun, LV and Foster, JM and Tzertzinis, G and Ono, M and Bandi, C and Slatko, BE and O'Neill, SL}, title = {Determination of Wolbachia genome size by pulsed-field gel electrophoresis.}, journal = {Journal of bacteriology}, volume = {183}, number = {7}, pages = {2219-2225}, pmid = {11244060}, issn = {0021-9193}, support = {AI40620/AI/NIAID NIH HHS/United States ; AI47409/AI/NIAID NIH HHS/United States ; }, mesh = {Chromosomes, Bacterial ; DNA, Bacterial/analysis ; Electrophoresis, Gel, Pulsed-Field ; *Genome, Bacterial ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Genome sizes of six different Wolbachia strains from insect and nematode hosts have been determined by pulsed-field gel electrophoresis of purified DNA both before and after digestion with rare-cutting restriction endonucleases. Enzymes SmaI, ApaI, AscI, and FseI cleaved the studied Wolbachia strains at a small number of sites and were used for the determination of the genome sizes of wMelPop, wMel, and wMelCS (each 1.36 Mb), wRi (1.66 Mb), wBma (1.1 Mb), and wDim (0.95 Mb). The Wolbachia genomes studied were all much smaller than the genomes of free-living bacteria such as Escherichia coli (4.7 Mb), as is typical for obligate intracellular bacteria. There was considerable genome size variability among Wolbachia strains, especially between the more parasitic A group Wolbachia infections of insects and the mutualistic C and D group infections of nematodes. The studies described here found no evidence for extrachromosomal plasmid DNA in any of the strains examined. They also indicated that the Wolbachia genome is circular.}, } @article {pmid11237467, year = {2001}, author = {Loppin, B and Berger, F and Couble, P}, title = {Paternal chromosome incorporation into the zygote nucleus is controlled by maternal haploid in Drosophila.}, journal = {Developmental biology}, volume = {231}, number = {2}, pages = {383-396}, doi = {10.1006/dbio.2000.0152}, pmid = {11237467}, issn = {0012-1606}, mesh = {Aneuploidy ; Animals ; Cell Movement ; Cell Nucleus/*metabolism/ultrastructure ; Chromosomes/*metabolism/ultrastructure ; Drosophila ; Female ; Fertilization/physiology ; Haploidy ; Heterochromatin/physiology ; Male ; Microscopy, Confocal ; *Mitosis ; Mutation ; Phenotype ; Sex Factors ; Time Factors ; Zygote/*physiology/ultrastructure ; }, abstract = {maternal haploid (mh) is a strict maternal effect mutation that causes the production of haploid gynogenetic embryos (eggs are fertilized but only maternal chromosomes participate in development). We conducted a cytological analysis of fertilization and early development in mh eggs to elucidate the mechanism of paternal chromosome elimination. In mh eggs, as in wild-type eggs, male and female pronuclei migrate and appose, the first mitotic spindle forms, and both parental sets of chromosomes congress on the metaphase plate. In contrast to control eggs, mh paternal sister chromatids fail to separate in anaphase of the first division. As a consequence the paternal chromatin stretches and forms a bridge in telophase. During the first three embryonic divisions, damaged paternal chromosomes are progressively eliminated from the spindles that organize around maternal chromosomes. A majority of mh embryos do not survive the deleterious presence of aneuploid nuclei and rapidly arrest their development. The rest of mh embryos develop as haploid gynogenetic embryos and die before hatching. The mh phenotype is highly reminiscent of the early developmental defects observed in eggs fertilized by ms(3)K81 mutant males and in eggs produced in incompatible crosses of Drosophila harboring the endosymbiont bacteria Wolbachia.}, } @article {pmid11228015, year = {2001}, author = {Bandi, C and Dunn, AM and Hurst, GD and Rigaud, T}, title = {Inherited microorganisms, sex-specific virulence and reproductive parasitism.}, journal = {Trends in parasitology}, volume = {17}, number = {2}, pages = {88-94}, doi = {10.1016/s1471-4922(00)01812-2}, pmid = {11228015}, issn = {1471-4922}, mesh = {Animals ; Arthropods/*microbiology/*parasitology ; Female ; Male ; Microsporidiosis/veterinary ; Reproduction ; Rickettsiaceae Infections/veterinary ; *Symbiosis ; Wolbachia/growth & development ; }, abstract = {Parasites show an amazing repertoire of adaptations, highlighted by complex life cycles that allow both survival in the host and transmission among hosts. However, there is one heterogeneous group of microorganisms whose adaptations are perhaps even more surprising: parthenogenesis induction, feminization of genetic males, killing of male hosts and sperm-mediated sterilization of uninfected eggs. The common feature of these microorganisms is their mode of transmission: inheritance from mother to offspring. Here, we present an introduction to hereditary symbiosis, focusing on microsporidia and bacteria that manipulate host reproduction in arthropods (reproductive parasites). We also discuss the implications of one of these microorganisms, Wolbachia, for the control of arthropod pests and vectors and for the therapy of filarial diseases. Finally, we discuss whether some parasites of vertebrates might show sex-specific virulence.}, } @article {pmid11217858, year = {2001}, author = {Bordenstein, SR and O'Hara, FP and Werren, JH}, title = {Wolbachia-induced incompatibility precedes other hybrid incompatibilities in Nasonia.}, journal = {Nature}, volume = {409}, number = {6821}, pages = {707-710}, doi = {10.1038/35055543}, pmid = {11217858}, issn = {0028-0836}, mesh = {Animals ; *Biological Evolution ; Crosses, Genetic ; Female ; Fertility ; Hybridization, Genetic ; Male ; Reproduction ; Species Specificity ; Sperm Motility ; Spermatozoa ; Symbiosis ; Wasps/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are cytoplasmically inherited bacteria that cause a number of reproductive alterations in insects, including cytoplasmic incompatibility, an incompatibility between sperm and egg that results in loss of sperm chromosomes following fertilization. Wolbachia are estimated to infect 15-20% of all insect species, and also are common in arachnids, isopods and nematodes. Therefore, Wolbachia-induced cytoplasmic incompatibility could be an important factor promoting rapid speciation in invertebrates, although this contention is controversial. Here we show that high levels of bidirectional cytoplasmic incompatibility between two closely related species of insects (the parasitic wasps Nasonia giraulti and Nasonia longicornis) preceded the evolution of other postmating reproductive barriers. The presence of Wolbachia severely reduces the frequency of hybrid offspring in interspecies crosses. However, antibiotic curing of the insects results in production of hybrids. Furthermore, F1 and F2 hybrids are completely viable and fertile, indicating the absence of F1 and F2 hybrid breakdown. Partial interspecific sexual isolation occurs, yet it is asymmetric and incomplete. Our results indicate that Wolbachia-induced reproductive isolation occurred in the early stages of speciation in this system, before the evolution of other postmating isolating mechanisms (for example, hybrid inviability and hybrid sterility).}, } @article {pmid11217844, year = {2001}, author = {Wade, MJ}, title = {Infectious speciation.}, journal = {Nature}, volume = {409}, number = {6821}, pages = {675-677}, doi = {10.1038/35055648}, pmid = {11217844}, issn = {0028-0836}, mesh = {Animals ; *Biological Evolution ; Cytoplasm ; Female ; Host-Parasite Interactions ; Male ; Reproduction ; Wasps/genetics/*parasitology ; Wolbachia/*physiology ; }, } @article {pmid11214099, year = {2000}, author = {Townson, S and Hutton, D and Siemienska, J and Hollick, L and Scanlon, T and Tagboto, SK and Taylor, MJ}, title = {Antibiotics and Wolbachia in filarial nematodes: antifilarial activity of rifampicin, oxytetracycline and chloramphenicol against Onchocerca gutturosa, Onchocerca lienalis and Brugia pahangi.}, journal = {Annals of tropical medicine and parasitology}, volume = {94}, number = {8}, pages = {801-816}, doi = {10.1080/00034980020027988}, pmid = {11214099}, issn = {0003-4983}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Brugia pahangi/*drug effects/ultrastructure ; Cattle ; Chloramphenicol/therapeutic use ; Female ; Filariasis/*drug therapy ; Gerbillinae ; Male ; Mice ; Mice, Inbred CBA ; Microscopy, Electron ; Onchocerciasis/*drug therapy ; Oxytetracycline/therapeutic use ; Rifampin/therapeutic use ; Treatment Outcome ; Wolbachia/*drug effects/ultrastructure ; }, abstract = {The activity against filarial parasites of the antibiotics rifampicin, oxytetracycline and chloramphenicol was examined. In addition, transmission electron microscopy was used to study the effects of rifampicin and oxytetracycline on filarial tissues and on the endosymbiont bacterium, Wolbachia. When tested in vitro at a concentration of 50.0 microM, each of the three antibiotics significantly reduced the motility levels of male Onchocerca gutturosa. Rifampicin, however, was the most active, virtually immobilizing the parasite by the end of the 40-day trial and producing an 84% reduction in viability (as measured by formazan-based colorimetry). In tests against O. lienalis microfilariae (mff) in CBA mice, the numbers of mff recovered after treatment with oxytetracycline at 100, 25 or 6.5 mg/kg daily, for 15 days, were 56% (P < or = 0.03), 38% (P> 0.05) and 45% (P = 0.05) less than that recovered from the untreated controls, respectively. In another trial in mice, rifampicin (100 mg/kg daily for 15 days) was found to be the most active (causing a 74% reduction in the number of mff recovered--approximately equal to that achieved with the positive control of a single dose of ivermectin at 2 microg/kg), with chloramphenicol also showing significant activity (39% reduction). In further, in-vivo trials, at three dose levels (100, 25 or 6.25 mg/kg daily, for 15 days), all three antibiotics were tested against adult Brugia pahangi in the peritoneal cavities of jirds. None of the antibiotics produced a significant reduction in the numbers of live worms recovered, although a marginal effect was observed in eight of the nine antibiotic-treated groups. A further extended trial with rifampicin and oxytetracycline resulted in 43% and 38% reductions in worm recoveries, respectively (not statistically significant but consistent with a marginal effect); some of these worms appeared less motile and qualitatively in poor condition compared with those recovered from untreated jirds. Ultrastructural studies of these treated worms revealed that virtually all of the endosymbiont bacteria had been cleared from the parasite tissues. The tissues of the adult worms appeared to be largely intact but with a granulomatous response of host cells adhering to some specimens. However, developing uterine forms appeared to be abnormal and extensively damaged, showing an abrogation of embryogenesis. In contrast, worms recovered from control animals contained large numbers of Wolbachia, had no adherent host cells, and showed normal ultrastructure; the female worms exhibited a full range of intra-uterine developing stages from eggs to stretched mff. It is likely that the activity of these antibiotics against the endosymbiont Wolbachia causes the observed antifilarial activity, although some direct effect of each drug on filarial viability cannot be ruled out.}, } @article {pmid11209779, year = {2000}, author = {Price, CS and Kim, CH and Posluszny, J and Coyne, JA}, title = {Mechanisms of conspecific sperm precedence in Drosophila.}, journal = {Evolution; international journal of organic evolution}, volume = {54}, number = {6}, pages = {2028-2037}, doi = {10.1111/j.0014-3820.2000.tb01246.x}, pmid = {11209779}, issn = {0014-3820}, support = {GM-07197/GM/NIGMS NIH HHS/United States ; R01 GM58260/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/anatomy & histology/microbiology/*physiology ; Female ; Genitalia, Male/anatomy & histology ; Male ; Reproduction ; Sexual Behavior, Animal/physiology ; Spermatozoa/microbiology/*physiology ; Wolbachia/pathogenicity ; }, abstract = {The postmating, prezygotic isolating mechanism known as conspecific sperm precedence (CSP) may play an important role in speciation, and understanding the mechanism of CSP is important in reconstructing its evolution. When a Drosophila simulans female mates with both a D. simulans male and a D. mauritiana male, the vast majority of her progeny are fathered by D. simulans, regardless of the order of mating. The dearth of hybrid progeny does not result from inviability of eggs fertilized by heterospecific sperm or from the relative inviability of heterospecific larvae. Instead, CSP apparently results from a prefertilization obstacle to heterospecific sperm. We identified two independent barriers to heterospecific fertilization, sperm displacement and incapacitation, whose action depends on the order of mating. When a D. simulans female mates first with a conspecific male, the seminal fluid from this mating incapacitates heterospecific sperm transferred two days later. This sperm incapacitation occurs with no change in the retention of stored sperm over time, but does not occur when the conspecific mating lasts for only 5 min. When the order of matings is reversed, the seminal fluid from the second mating physically displaces heterospecific sperm from storage, even if the conspecific copulation lasts only 5 min. Conspecific sperm are not susceptible to displacement by a second conspecific copulation, but are susceptible to interference by heterospecific sperm if the conspecific copulation is interrupted after 12 min. Curing the D. mauritiana males of their infection with the endosymbiont Wolbachia had no effect on CSP. Sperm displacement and incapacitation involve the same basic mechanisms seen in second-male sperm precedence within species, supporting the hypothesis that CSP is an evolutionary by-product of adaptations affecting sperm competition within species.}, } @article {pmid11201358, year = {2000}, author = {van der Geest, LP and Elliot, SL and Breeuwer, JA and Beerling, EA}, title = {Diseases of mites.}, journal = {Experimental & applied acarology}, volume = {24}, number = {7}, pages = {497-560}, pmid = {11201358}, issn = {0168-8162}, mesh = {Animals ; Humans ; *Mites/microbiology/parasitology/virology ; }, abstract = {An overview is given of studies on diseases of mites. Knowledge of diseases of mites is still fragmentary but in recent years more attention has been paid to acaropathogens, often because of the economic importance of many mite species. Most research on mite pathogens concerns studies on fungal pathogens of eriophyoids and spider mites especially. These fungi often play an important role in the regulation of natural mite populations and are sometimes able to decimate populations of phytophagous mites. Studies are being conducted to develop some of these fungi as commercial acaricides. Virus diseases are known in only a few mites, namely, the citrus red mite and the European red mite. In both cases, non-occluded viruses play an important role in the regulation of mite populations in citrus and peach orchards, respectively, but application of these viruses as biological control agents does not seem feasible. A putative iridovirus has been observed in association with Varroa mites in moribund honeybee colonies. The virus is probably also pathogenic for honeybees and may be transmitted to them through this parasitic mite. Few bacteria have been reported as pathogens of the Acari but in recent years research has been concentrated on intracellular organisms such as Wolbachia that may cause distorted sex ratios in offspring and incompatibility between populations. The role of these organisms in natural populations of spider mites is in particular discussed. The effect of Bacillus thuringiensis on mites is also treated in this review, although its mode of action in arthropods is mainly due to the presence of toxins and it is, therefore, not considered to be a pathogen in the true sense of the word. Microsporidia have been observed in several mite species especially in oribatid mites, although other groups of mites may also be affected. In recent years, Microsporidia infections in Phytoseiidae have received considerable attention, as they are often found in mass rearings of beneficial arthropods. They affect the efficacy of these predators as biological control agent of insect and mite pests. Microsporidia do not seem to have potential for biological control of mites.}, } @article {pmid11198638, year = {2000}, author = {Taylor, MJ}, title = {Wolbachia bacteria of filarial nematodes in the pathogenesis of disease and as a target for control.}, journal = {Transactions of the Royal Society of Tropical Medicine and Hygiene}, volume = {94}, number = {6}, pages = {596-598}, doi = {10.1016/s0035-9203(00)90201-3}, pmid = {11198638}, issn = {0035-9203}, mesh = {Animals ; Anti-Bacterial Agents/*therapeutic use ; Elephantiasis, Filarial/*prevention & control ; Humans ; Pest Control, Biological/*methods ; Public Health ; *Rickettsiaceae ; Symbiosis ; Wuchereria bancrofti/*microbiology ; }, } @article {pmid11197770, year = {2001}, author = {Casiraghi, M and Anderson, TJ and Bandi, C and Bazzocchi, C and Genchi, C}, title = {A phylogenetic analysis of filarial nematodes: comparison with the phylogeny of Wolbachia endosymbionts.}, journal = {Parasitology}, volume = {122 Pt 1}, number = {}, pages = {93-103}, doi = {10.1017/s0031182000007149}, pmid = {11197770}, issn = {0031-1820}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/genetics ; *Cytoskeletal Proteins ; Electron Transport Complex IV/genetics ; Filarioidea/*classification/enzymology/genetics/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/chemistry ; Symbiosis ; Thelazioidea/classification/enzymology/genetics ; Wolbachia/*classification/enzymology ; }, abstract = {Infection with the endosymbiotic bacteria Wolbachia is widespread in filarial nematodes. Previous studies have suggested concordance between the phylogeny of Wolbachia with that of their nematode hosts. However, there is only one published molecular phylogenetic study of filarial species, based on the 5S rRNA gene spacer. The phylogeny proposed by this study is partially incongruent with previous classifications of filarial nematodes, based on morphological characters. Furthermore, both traditional classifications and molecular phylogenies are, in part, inconsistent with the phylogeny of Wolbachia. Here we report mitochondrial cytochrome oxidase I (COI) gene sequences for 11 species of filaria and for another spirurid nematode which was included as an outgroup. In addition, 16S rRNA, wsp and ftsZ gene sequences were generated for the Wolbachia of several filarial species, in order to complete the available data sets and further resolve the phylogeny of Wolbachia in nematodes. We used these data to evaluate whether nematode and Wolbachia phylogenies are concordant. Some of the possible phylogenetic reconstructions based on COI gene were congruent with the phylogeny of Wolbachia and supported the grouping of the rodent filaria Litomosoides sigmodontis with the lymphatic filariae (i.e. Brugia spp. and Wuchereria spp.) and the sister group relationship of Dirofilaria spp. and Onchocerca spp. However, the placement of the Wolbachia-free filaria Acanthocheilonema viteae is ambiguous and dependent on the phylogenetic methods used.}, } @article {pmid11197127, year = {2000}, author = {Bazzocchi, C and Ceciliani, F and McCall, JW and Ricci, I and Genchi, C and Bandi, C}, title = {Antigenic role of the endosymbionts of filarial nematodes: IgG response against the Wolbachia surface protein in cats infected with Dirofilaria immitis.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1461}, pages = {2511-2516}, pmid = {11197127}, issn = {0962-8452}, mesh = {Amino Acid Sequence ; Animals ; Antigens, Bacterial/genetics/*immunology ; Base Sequence ; Blotting, Western ; Cat Diseases/immunology/*parasitology ; Cats ; Dirofilaria immitis/*microbiology ; Dirofilariasis/parasitology ; Immunoglobulin G/blood ; Membrane Proteins/genetics/immunology ; Molecular Sequence Data ; Sequence Alignment ; *Symbiosis ; Wolbachia/*immunology ; }, abstract = {Filarial nematodes harbour intracellular endosymbiotic bacteria, which have been assigned to the genus Wolbachia. These bacteria appear to play an important role in the pathogenesis of filarial diseases through their lipopolysaccharides. In view of the presence of Wolbachia endosymbionts in the body of filarial nematodes, one might also expect that proteins from these bacteria play an antigenic role in humans and animals affected by filariases. To test this hypothesis, we produced in recombinant form the surface protein WSP and a portion of the cell-cycle protein FTSZ from the Wolbachia of Dirofilaria immitis. Western immunoblot assays were then performed using cat sera to test the immunogenicity of these proteins. Sera were collected from owners' cats, which were either sero-negative or sero-positive for D. immitis and from cats before and after experimental infection with D. immitis. FTSZ was recognized in Western blots by sera from both positive and negative cats and from both uninfected and experimentally infected cats. WSP was recognized only by sera from positive cats and from cats experimentally infected with D. immitis; this protein was not recognized by sera from negative cats and from cats before experimental infection with D. immitis. The results of Western blot assays on WSP thus support the hypothesis that infection with filarial nematodes induces the production of antibodies against Wolbachia proteins.}, } @article {pmid11195346, year = {2000}, author = {Beukeboom, LW and Pijnacker, LP}, title = {Automictic parthenogenesis in the parasitoid Venturia canescens (Hymenoptera: Ichneumonidae) revisited.}, journal = {Genome}, volume = {43}, number = {6}, pages = {939-944}, doi = {10.1139/g00-061}, pmid = {11195346}, issn = {0831-2796}, mesh = {Animals ; Chromosomes ; Hymenoptera/classification/*genetics/microbiology ; Meiosis/genetics ; *Parthenogenesis ; Polymerase Chain Reaction ; Wolbachia/genetics/isolation & purification ; }, abstract = {Both arrhenotokous and thelytokous reproduction are known to occur in the parasitoid wasp Venturia canescens. The cytological mechanism of thelytoky was previously reported to involve the formation of a restitution metaphase after the reduction division, but the exact nature of the subsequent divisions, whether reductional or equational, remained unclear. We reinvestigated the cytological mechanisms in a thelytokous strain collected in France. Our observations confirm previous results, but an equational and not a reduction division was observed after restitution. This type of reproduction can be classified as central fusion automictic parthenogenesis. In two arrhenotokous strains the normal pattern of oogenesis and syngamy of Hymenoptera was observed. In addition, we used PCR amplification to show that thelytoky in V. canescens is not caused by Wolbachia bacteria. The results are discussed in relation to maintenance of heterozygosity and female sex.}, } @article {pmid11183760, year = {2000}, author = {Enserink, M}, title = {Mosquito engineering. Building a disease-fighting mosquito.}, journal = {Science (New York, N.Y.)}, volume = {290}, number = {5491}, pages = {440-441}, doi = {10.1126/science.290.5491.440}, pmid = {11183760}, issn = {0036-8075}, mesh = {Aedes/*genetics/immunology/parasitology ; Animals ; Anopheles/*genetics/immunology/parasitology ; Antibodies, Protozoan/biosynthesis ; *DNA Transposable Elements ; Defensins/genetics/metabolism ; Gene Transfer Techniques ; *Genetic Engineering ; Insect Vectors/*genetics/growth & development/immunology ; Malaria/*prevention & control/transmission ; Plasmodium/growth & development/immunology ; Wolbachia/physiology ; }, abstract = {With resistance against insecticides on the rise and a U.N.-backed push to phase out DDT, several labs have embarked on the most ambitious and futuristic of all approaches to combat malaria: They hope to replace billions and billions of mosquitoes in the world's endemic areas with new strains, created in the lab, that would be "refractory," or unable to transmit Plasmodium, the parasite that causes the disease. The idea is not that farfetched, these researchers claim.}, } @article {pmid11155997, year = {2000}, author = {Vandekerckhove, TT and Willems, A and Gillis, M and Coomans, A}, title = {Occurrence of novel verrucomicrobial species, endosymbiotic and associated with parthenogenesis in Xiphinema americanum-group species (Nematoda, Longidoridae).}, journal = {International journal of systematic and evolutionary microbiology}, volume = {50 Pt 6}, number = {}, pages = {2197-2205}, doi = {10.1099/00207713-50-6-2197}, pmid = {11155997}, issn = {1466-5026}, mesh = {Animals ; DNA, Ribosomal/analysis ; Female ; Gram-Negative Bacteria/*classification/genetics/physiology ; Indoles ; Microscopy, Electron ; Microscopy, Fluorescence/methods ; Molecular Sequence Data ; Nematoda/*microbiology/physiology ; Parthenogenesis/*physiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Numerous micro-organisms have been described as cytoplasmic symbionts of eukaryotes. Many so-called obligate endosymbionts rely exclusively on maternal (vertical or transovarial) transmission to maintain themselves, rendering them dependent on the host sex ratio, which they would tend to manipulate to their own advantage. The latter phenomenon is often associated with the presence of Wolbachia pipientis (alpha-Proteobacteria) in arthropods and nematodes. A potentially similar situation was discovered involving members of a new clade of Verrucomicrobia, another main line of descent in the Bacteria. Nematode species of the Xiphinema americanum group (Nematoda, Longidoridae), viz. Xiphinema americanum, Xiphinema rivesi and Xiphinema brevicollum, each harbour their own specific verrucomicrobial endosymbionts. They are exclusively maternally inherited and their hosts reproduce by thelytokous (mother-to-daughter) parthenogenesis, males being extremely rare. A new genus, 'Candidatus Xiphinematobacter' gen. nov., along with three new candidate verrucomicrobial species, 'Candidatus Xiphinematobacter americani' sp. nov., 'Candidatus Xiphinematobacter rivesi' sp. nov. and 'Candidatus Xiphinematobacter brevicolli' sp. nov., are described on the basis of transmission electron microscopy, scanning electron microscopy, DAPI (4',6-diamidino-2-phenylindole) epifluorescence microscopy and 16S rDNA sequence analysis. These are the first endosymbiotic species described among the Verrucomicrobia. They share a mean 16S rDNA similarity of about 93%, whereas similarity to their closest relative, clone WCHD3-88, is less than 87%. Thus, the endosymbionts form a homogeneous clade for which the new candidate genus 'Candidatus Xiphinematobacter' gen. nov. is proposed. The type species is 'Candidatus Xiphinematobacter brevicolli' sp. nov.}, } @article {pmid11128800, year = {2000}, author = {Fleury, F and Vavre, F and Ris, N and Fouillet, P and Boulétreau, M}, title = {Physiological cost induced by the maternally-transmitted endosymbiont Wolbachia in the Drosophila parasitoid Leptopilina heterotoma.}, journal = {Parasitology}, volume = {121 Pt 5}, number = {}, pages = {493-500}, doi = {10.1017/s0031182099006599}, pmid = {11128800}, issn = {0031-1820}, mesh = {Animals ; Drosophila melanogaster/*parasitology ; Female ; Fertility/physiology ; Host-Parasite Interactions ; Image Processing, Computer-Assisted ; Male ; Motor Activity/physiology ; Sex Ratio ; Videotape Recording ; Wasps/growth & development/*microbiology/*physiology ; Wolbachia/*pathogenicity ; }, abstract = {Endosymbiotic bacteria of the genus Wolbachia infect a number of invertebrate species in which they induce various alterations in host reproduction, mainly cytoplasmic incompatibility (CI). In contrast to most other maternally transmitted parasites, manipulation of host reproduction makes the spread of Wolbachia possible even if they induce a physiological cost on their hosts. Current studies have shown that fitness consequences of Wolbachia infection could range from positive (mutualist) to negative (parasitic) but, in most cases, Wolbachia do not have strong deleterious effects on host fitness and the status of association remains unclear. Here, we show that in the Drosophila parasitoid wasp Leptopilina heterotoma, Wolbachia infection has a negative impact on several host fitness traits of both sexes. Fecundity, adult survival and locomotor performance are significantly reduced, whereas circadian rhythm, development time and offspring sex-ratio are not affected. Although the cost of bacterial infection can be overcome by effects on host reproduction i.e. cytoplasmic incompatibility, it could influence the spread of the bacterium at the early stages of the invasion process. Clearly, results underline the wide spectrum of phenotypic effects of Wolbachia infection and, to our knowledge, Wolbachia infection of L. heterotoma appears to be one of the most virulent that has ever been observed in insects.}, } @article {pmid11122476, year = {2000}, author = {Shoemaker, DD and Ross, KG and Keller, L and Vargo, EL and Werren, JH}, title = {Wolbachia infections in native and introduced populations of fire ants (Solenopsis spp.).}, journal = {Insect molecular biology}, volume = {9}, number = {6}, pages = {661-673}, doi = {10.1046/j.1365-2583.2000.00233.x}, pmid = {11122476}, issn = {0962-1075}, mesh = {Animals ; Ants/classification/genetics/growth & development/*microbiology ; DNA, Bacterial ; DNA, Mitochondrial ; Genetic Variation ; Phylogeny ; South America ; Species Specificity ; United States ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Wolbachia are cytoplasmically inherited bacteria that induce a variety of effects with fitness consequences on host arthropods, including cytoplasmic incompatibility, parthenogenesis, male-killing and feminization. We report here the presence of Wolbachia in native South American populations of the fire ant Solenopsis invicta, but the apparent absence of the bacteria in introduced populations of this pest species in the USA. The Wolbachia strains in native S. invicta are of two divergent types (A and B), and the frequency of infection varies dramatically between geographical regions and social forms of this host. Survey data reveal that Wolbachia also are found in other native fire ant species within the Solenopsis saevissima species complex from South America, including S. richteri. This latter species also has been introduced in the USA, where it lacks Wolbachia. Sequence data reveal complete phylogenetic concordance between mtDNA haplotype in S. invicta and Wolbachia infection type (A or B). In addition, the mtDNA and associated group A Wolbachia strain in S. invicta are more closely related to the mtDNA and Wolbachia strain found in S. richteri than they are to the mtDNA and associated group B Wolbachia in S. invicta. These data are consistent with historical introgression of S. richteri cytoplasmic elements into S. invicta populations, resulting in enhanced infection and mtDNA polymorphisms in S. invicta. Wolbachia may have significant fitness effects on these hosts (either directly or by cytoplasmic incompatibility) and therefore these microbes potentially could be used in biological control programmes to suppress introduced fire ant populations.}, } @article {pmid11122472, year = {2000}, author = {Bourtzis, K and Pettigrew, MM and O'Neill, SL}, title = {Wolbachia neither induces nor suppresses transcripts encoding antimicrobial peptides.}, journal = {Insect molecular biology}, volume = {9}, number = {6}, pages = {635-639}, doi = {10.1046/j.1365-2583.2000.00224.x}, pmid = {11122472}, issn = {0962-1075}, support = {AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/immunology/*microbiology ; Amino Acid Sequence ; Animals ; Anti-Bacterial Agents/metabolism ; Base Sequence ; Cloning, Molecular ; DNA ; Defensins/genetics ; Drosophila/genetics/immunology/*microbiology ; Gene Expression Regulation ; Genes, Insect ; Genetic Markers ; Molecular Sequence Data ; Sequence Alignment ; Transcription, Genetic ; Wolbachia/immunology/*physiology ; }, abstract = {Wolbachia are intracellular maternally inherited microorganisms that are associated with reproductive abnormalities such as cytoplasmic incompatibility (CI), feminization and parthenogenesis in the various arthropod species they infect. Surveys indicate that these bacteria infect more than 16% of all insect species as well as isopods, mites and nematodes, making Wolbachia one of the most ubiquitous parasites yet described. However, nothing is known about the interactions of this bacterium with the host's immune system. We studied the expression of inducible antimicrobial markers in the adults of two Wolbachia infected insect species, Drosophila simulans and Aedes albopictus. The lack of available immune markers in the mosquito species led us to clone part of the defensin gene from this species, which was found to be very similar to the other mosquito defensins cloned from Anopheles gambiae and Aedes aegypti. Comparisons of the expression pattern of the antibacterial markers between Wolbachia-infected and cured lines, and also between bacteria-challenged and unchallenged adults indicated that Wolbachia does not either constitutively induce or suppress the transcription of these antibacterial genes. In addition, no difference in the transcription of these genes was found between double and single Wolbachia-infected strains or between strains in which Wolbachia has different tissue tropisms.}, } @article {pmid11108593, year = {2000}, author = {James, AC and Ballard, JW}, title = {Expression of cytoplasmic incompatibility in Drosophila simulans and its impact on infection frequencies and distribution of Wolbachia pipientis.}, journal = {Evolution; international journal of organic evolution}, volume = {54}, number = {5}, pages = {1661-1672}, doi = {10.1111/j.0014-3820.2000.tb00710.x}, pmid = {11108593}, issn = {0014-3820}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm/genetics ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Drosophila/genetics/*microbiology/*physiology ; Drosophila melanogaster/microbiology ; Female ; Genetics, Population ; Madagascar ; Male ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Reunion ; Wolbachia/classification/genetics/*physiology ; }, abstract = {The aim of this study is to examine the expression of cytoplasmic incompatibility and investigate the distribution and population frequencies of Wolbachia pipientis strains in Drosophila simulans. Nucleotide sequence data from 16S rDNA and a Wolbachia surface protein coding sequence and cytoplasmic incompatibility assays identify four distinct Wolbachia strains: wHa, wRi, wMa, and wAu. The levels of cytoplasmic incompatibility between six lines carrying these strains of bacteria and three control lines without bacteria are characterized. Flies infected with wHa and wRi are bidirectionally incompatible, and males that carry either strain can only successfully produce normal numbers of offspring with females carrying the same bacterial strain. Males infected with wAu do not express incompatibility. Males infected with the wMa strain express intermediate incompatibility when mated to females with no bacteria and no incompatibility with females with any other Wolbachia strain. We conduct polymerase chain reaction/restriction fragment length polymorphism assays to distinguish the strain of Wolbachia and the mitochondrial haplotype to survey populations for each type and associations between them. Drosophila simulans is known to have three major mitochondrial haplotypes (siI, sill, and siIII) and two subtypes (siIIA and siIIB). All infected lines of the sil haplotype carry wHa, wNo, or both; wMa and wNo are closely related and it is not clear whether they are distinct strains or variants of the same strain. Infected lines with the silIA haplotype harbor wRi and the siIIB haplotype carries wAu. The wMa infection is found in siIII haplotype lines. The phenotypic expression of cytoplasmic incompatibility and its relation to between-population differences in frequencies of Wolbachia infection are discussed.}, } @article {pmid11080372, year = {2000}, author = {Masui, S and Kamoda, S and Sasaki, T and Ishikawa, H}, title = {Distribution and evolution of bacteriophage WO in Wolbachia, the endosymbiont causing sexual alterations in arthropods.}, journal = {Journal of molecular evolution}, volume = {51}, number = {5}, pages = {491-497}, doi = {10.1007/s002390010112}, pmid = {11080372}, issn = {0022-2844}, mesh = {Amino Acid Sequence ; Animals ; Arthropods/genetics/*virology ; Bacteriophages/*genetics ; Cloning, Molecular ; DNA, Bacterial/chemistry/genetics ; DNA, Viral/chemistry/genetics ; *Evolution, Molecular ; Gene Expression Regulation, Viral ; Genes, Viral/genetics ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Sex Determination Processes ; Symbiosis/genetics ; Wolbachia/*genetics/virology ; }, abstract = {Wolbachia are obligatory intracellular and maternally inherited bacteria, known to infect many species of arthropod. In this study, we discovered a bacteriophage-like genetic element in Wolbachia, which was tentatively named bacteriophage WO. The phylogenetic tree based on phage WO genes of several Wolbachia strains was not congruent with that based on chromosomal genes of the same strains, suggesting that phage WO was active and horizontally transmitted among various Wolbachia strains. All the strains of Wolbachia used in this study were infected with phage WO. Although the phage genome contained genes of diverse origins, the average G+C content and codon usage of these genes were quite similar to those of a chromosomal gene of Wolbachia. These results raised the possibility that phage WO has been associated with Wolbachia for a very long time, conferring some benefit to its hosts. The evolution and possible roles of phage WO in various reproductive alterations of insects caused by Wolbachia are discussed.}, } @article {pmid11075704, year = {2000}, author = {Vala, F and Breeuwer, JA and Sabelis, MW}, title = {Wolbachia-induced 'hybrid breakdown' in the two-spotted spider mite Tetranychus urticae Koch.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1456}, pages = {1931-1937}, pmid = {11075704}, issn = {0962-8452}, mesh = {Animals ; Crosses, Genetic ; Female ; Male ; Mites/*microbiology/physiology ; Reproduction/*physiology ; Sex Ratio ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The most common post-zygotic isolation mechanism between populations of the phytophagous mite Tetranychus urticae is 'hybrid breakdown', i.e. when individuals from two different populations are crossed, F1 hybrid females are produced, but F2 recombinant male offspring suffer increased mortality. Two-spotted spider mites collected from two populations, one on rose and the other on cucumber plants, were infected with Wolbachia bacteria. These bacteria may induce cytoplasmic incompatibility in their hosts: uninfected (U) females become reproductively incompatible with infected (W) males. We report on the effect of Wolbachia infections in intra- and interstrain crosses on (i) F1 mortality and sex ratios (a test for cytoplasmic incompatibility), and (ii) the number of haploid offspring and mortality in clutches of F1 virgins (a test for hybrid breakdown). U x W crosses within the rose strain exhibited partial cvtoplasmic incompatibility. More interestingly, F2 males suffered increased mortality, a result identical to the hybrid breakdown phenomenon. The experiments were repeated using females from the cucumber strain. In interstrain U x W and U x U crosses, hybrid breakdown was much stronger in the former (80 versus 26%). This is the first report of a Wolbachia infection causing a hybrid breakdown phenotype. Our results show that Wolbhachia infections can contribute to reproductive incompatibility between populations of T. urticae.}, } @article {pmid11053403, year = {2000}, author = {Masui, S and Sasaki, T and Ishikawa, H}, title = {Genes for the type IV secretion system in an intracellular symbiont, Wolbachia, a causative agent of various sexual alterations in arthropods.}, journal = {Journal of bacteriology}, volume = {182}, number = {22}, pages = {6529-6531}, pmid = {11053403}, issn = {0021-9193}, mesh = {Animals ; Arthropods/*microbiology/physiology ; Bacterial Proteins/*genetics/metabolism ; *Genes, Bacterial ; Molecular Sequence Data ; Operon ; Reproduction/physiology ; Symbiosis ; *Virulence Factors ; Wolbachia/*genetics/metabolism ; }, abstract = {Wolbachia species are intracellular bacteria known to cause reproductive abnormalities in their hosts. In this study, we identified Wolbachia genes encoding homologs to the type IV secretion system by which many pathogenic bacteria secrete macromolecules. The genes identified encoded most of the essential components of the secretion system and were cotranscribed as an operon.}, } @article {pmid11041802, year = {2000}, author = {Cook, JM and Rokas, A}, title = {Influential passengers come of age. 1st International Wolbachia Conference, Orthodox Academy, Kolymbari, Crete, Greece, 7-12 June 2000.}, journal = {Trends in genetics : TIG}, volume = {16}, number = {9}, pages = {378-379}, doi = {10.1016/s0168-9525(00)02085-0}, pmid = {11041802}, issn = {0168-9525}, mesh = {Humans ; *Wolbachia/isolation & purification/physiology ; }, } @article {pmid11040288, year = {2000}, author = {Pintureau, B and Chaudier, S and Lassablière, F and Charles, H and Grenier, S}, title = {Addition of wsp sequences to the Wolbachia phylogenetic tree and stability of the classification.}, journal = {Journal of molecular evolution}, volume = {51}, number = {4}, pages = {374-377}, doi = {10.1007/s002390010099}, pmid = {11040288}, issn = {0022-2844}, mesh = {Animals ; Asia ; Bacterial Outer Membrane Proteins/*genetics ; DNA Primers ; Europe ; *Phylogeny ; Polymerase Chain Reaction/methods ; South America ; Species Specificity ; Symbiosis ; Wasps/microbiology ; *Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia are symbiotic bacteria altering reproductive characters of numerous arthropods. Their most recent phylogeny and classification are based on sequences of the wsp gene. We sequenced wsp gene from six Wolbachia strains infecting six Trichogramma species that live as egg parasitoids on many insects. This allows us to test the effect of the addition of sequences on the Wolbachia phylogeny and to check the classification of Wolbachia infecting Trichogramma. The six Wolbachia studied are classified in the B supergroup. They confirm the monophyletic structure of the B Wolbachia in Trichogramma but introduce small differences in the Wolbachia classification. Modifications include the definition of a new group, Sem, for Wolbachia of T. semblidis and the merging of the two closely related groups, Sib and Kay. Specific primers were determined and tested for the Sem group.}, } @article {pmid11038309, year = {2000}, author = {Smith, HL and Rajan, TV}, title = {Tetracycline inhibits development of the infective-stage larvae of filarial nematodes in vitro.}, journal = {Experimental parasitology}, volume = {95}, number = {4}, pages = {265-270}, doi = {10.1006/expr.2000.4525}, pmid = {11038309}, issn = {0014-4894}, support = {AI39705/AI/NIAID NIH HHS/United States ; AI42362/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia malayi/*drug effects/growth & development/microbiology ; Brugia pahangi/*drug effects/growth & development/microbiology ; DNA, Bacterial/chemistry/isolation & purification ; Dirofilaria immitis/*drug effects/growth & development/microbiology ; Larva/drug effects/microbiology/physiology ; Molting/drug effects ; Tetracycline/*pharmacology ; Wolbachia/drug effects/genetics/physiology ; }, abstract = {In recent years, studies have linked tetracycline treatment of filaria-infected animals with reduced adult worm burdens and decreased levels of microfilaremia. These observations are believed to be attributable to clearance of Wolbachia, intracellular rickettsial-like organisms found within filarial tissues. Although maximal worm reductions were observed when treatment was initiated early in infection, it is not known whether tetracycline inhibits development of infective-stage larvae. To address this issue, we studied the effect of tetracycline on three different species of filarial nematodes, Brugia malayi, Brugia pahangi, and Dirofilaria immitis, in a serumfree in vitro system supporting molting to the fourth larval stage. Tetracycline was capable of inhibiting L3 to L4 molting within a dosage range similar to that reported for susceptible rickettsial organisms. However, Wolbachia DNA could still be detected in nematodes from tetracycline-treated cultures. In addition, three other antibiotics with anti-rickettsial and anti-chlamydial activity (chloramphenicol, erythromycin, and ciprofloxacin) failed to inhibit L3 to L4 molting. Although tetracycline is capable of completely blocking molting of infective-stage larvae, it remains possible that this effect is due to pharmacological activities unrelated to its anti-rickettsial functions.}, } @article {pmid11029673, year = {2000}, author = {Oh, HW and Kim, MG and Shin, SW and Bae, KS and Ahn, YJ and Park, HY}, title = {Ultrastructural and molecular identification of a Wolbachia endosymbiont in a spider, Nephila clavata.}, journal = {Insect molecular biology}, volume = {9}, number = {5}, pages = {539-543}, doi = {10.1046/j.1365-2583.2000.00218.x}, pmid = {11029673}, issn = {0962-1075}, mesh = {Animals ; Bacterial Proteins/genetics ; *Cytoskeletal Proteins ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Female ; Genes, Bacterial ; Microscopy, Electron ; Molecular Sequence Data ; Ovum/microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Spiders/*microbiology ; *Symbiosis ; Wolbachia/*classification/genetics/*ultrastructure ; }, abstract = {Wolbachia-like bacteria were observed in the egg cells of golden orb-weaving spider, Nephila clavata, by means of transmission electron microscopy. The bacteria exhibited the typical morphology of Wolbachia, including three enveloping membranes. Based on the amplification and sequencing of partial 16S rDNA and ftsZ gene, the bacteria were identified as Wolbachia, intracellular, transovarially inherited alpha-proteobacteria in invertebrates. Phylogenetic analysis based on 16S rDNA and ftsZ gene sequences invariably indicated that the intracellular bacteria from N. clavata belonged to group A Wolbachia, which were found only from insects. Clustering of Wolbachia from N. clavata with group A Wolbachia indicates that the bacteria were probably transferred horizontally between insects and the spider.}, } @article {pmid11014817, year = {2000}, author = {Hurst, GD and Johnson, AP and Schulenburg, JH and Fuyama, Y}, title = {Male-killing Wolbachia in Drosophila: a temperature-sensitive trait with a threshold bacterial density.}, journal = {Genetics}, volume = {156}, number = {2}, pages = {699-709}, pmid = {11014817}, issn = {0016-6731}, mesh = {Animals ; Crosses, Genetic ; Drosophila/classification/*genetics/*microbiology ; Female ; Fertility ; Larva ; Male ; Phenotype ; Phylogeny ; Rifampin/pharmacology ; Sex Ratio ; Temperature ; Wolbachia/drug effects/growth & development/*pathogenicity ; }, abstract = {Inherited microorganisms that disturb the reproduction of their host have been characterized from a number of host taxa. To understand the general principles underlying the genetic and mechanistic basis of interactions, study of different agents in model host species is required. To this end, the nature and genetics of the maternally inherited sex-ratio trait of Drosophila bifasciata were investigated. Successful curing of affected lines with antibiotics demonstrated this trait was associated with the presence of a bacterium, and molecular systematic analysis demonstrated an association between the presence of the trait and infection with an A group Wolbachia. The penetrance and heritability of the trait did not vary with maternal age. Exposure to elevated temperatures did reduce trait penetrance but did not affect heritability. Examination of the effect of temperature on bacterial density in eggs revealed a decrease in bacterial density following exposure of the parent to elevated temperature, consistent with the hypothesis that male killing in D. bifasciata requires a threshold density of Wolbachia within eggs. The male offspring produced following exposure to elevated temperatures were infected with Wolbachia on emergence as adults. Crossing studies demonstrated a weak cytoplasmic incompatibility phenotype exhibited by Wolbachia in these males. The results are discussed with respect to the incidence of male killing within the clade Wolbachia, the general nature of Wolbachia-host interactions, and the prospects for using this association to investigate the mechanism of male killing.}, } @article {pmid11012722, year = {2000}, author = {Poinsot, D and Montchamp-Moreau, C and Merçot, H}, title = {Wolbachia segregation rate in Drosophila simulans naturally bi-infected cytoplasmic lineages.}, journal = {Heredity}, volume = {85 (Pt 2)}, number = {}, pages = {191-198}, doi = {10.1046/j.1365-2540.2000.00736.x}, pmid = {11012722}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm/microbiology ; Drosophila/*microbiology ; Female ; Male ; Species Specificity ; Wolbachia/classification/*physiology ; }, abstract = {Wolbachia are maternally transmitted endocellular bacteria infecting several arthropod species. In order to study Wolbachia segregation rate, Drosophila simulans females from an Indo-Pacific population (Seychelles) bi-infected by the two Wolbachia variants wHa and wNo were backcrossed to uninfected males in two conditions. In the first case, Seychelles males from a stock cured from its Wolbachia by tetracycline treatment were used. In the second case, the males came from a naturally uninfected Tunisian population. It was found that (i) the two Wolbachia variants can segregate, so that bi-infected females can produce a few offspring infected only by wHa or wNo. This occurs in both backcross conditions. (ii) Segregation leads more frequently to wHa than to wNo mono-infection. (iii) Wolbachia transmission is lower when the Seychelles genome is introgressed by the Tunisian genome, suggesting that host genomic factors might influence infection fate.}, } @article {pmid11012714, year = {2000}, author = {Sasaki, T and Ishikawa, H}, title = {Transinfection of Wolbachia in the mediterranean flour moth, Ephestia kuehniella, by embryonic microinjection.}, journal = {Heredity}, volume = {85 (Pt 2)}, number = {}, pages = {130-135}, doi = {10.1046/j.1365-2540.2000.00734.x}, pmid = {11012714}, issn = {0018-067X}, mesh = {Animals ; Crosses, Genetic ; Cytoplasm ; Female ; Male ; Microinjections ; Moths/embryology/*microbiology ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Wolbachia are maternally transmitted intracellular bacteria found in many arthropod species. They cause a reproductive incompatibility called cytoplasmic incompatibility (CI) in several hosts, including the Mediterranean flour moth, Ephestia kuehniella. Two strains of E. kuehniella, one from Yokohama city and the other from Tsuchiura city, express different levels of CI: the Yokohama strain expresses CI at a higher level than the Tsuchiura strain. In order to determine whether the difference of CI levels depends on Wolbachia or the host, we performed transinfection experiments in E. kuehniella by means of embryonic microinjection, and successfully transferred Wolbachia carried by the Yokohama strain into the Tsuchiura strain, from which the original Wolbachia had been removed by tetracycline treatment. The resulting transinfected strain expressed CI at a level near that of the Yokohama strain, suggesting that, in these strains of E. kuehniella, the level of CI is determined by Wolbachia rather than by the host.}, } @article {pmid11008105, year = {2000}, author = {Brattig, NW and Rathjens, U and Ernst, M and Geisinger, F and Renz, A and Tischendorf, FW}, title = {Lipopolysaccharide-like molecules derived from Wolbachia endobacteria of the filaria Onchocerca volvulus are candidate mediators in the sequence of inflammatory and antiinflammatory responses of human monocytes.}, journal = {Microbes and infection}, volume = {2}, number = {10}, pages = {1147-1157}, doi = {10.1016/s1286-4579(00)01269-7}, pmid = {11008105}, issn = {1286-4579}, mesh = {Animals ; Antigens, CD/metabolism ; B7-1 Antigen/metabolism ; B7-2 Antigen ; Cells, Cultured ; Female ; HLA-DR Antigens/metabolism ; Humans ; Inflammation Mediators/*metabolism/pharmacology ; Interleukin-10/biosynthesis ; Lipopolysaccharides/*metabolism/pharmacology ; Membrane Glycoproteins/metabolism ; Monocytes/*metabolism/parasitology ; Onchocerca volvulus/metabolism/*microbiology ; Onchocerciasis/*metabolism/pathology ; Tumor Necrosis Factor-alpha/biosynthesis ; Wolbachia/*metabolism ; }, abstract = {The majority of Onchocerca volvulus-infected persons show signs of cellular anergy, and long-time survival of adult and larval parasites in subcutaneous tissue is observed. The mechanisms leading to immunological hyporesponsiveness are poorly understood. Monocytes/macrophages represent a link between the innate and acquired immune system and are candidate cells to promote inflammatory and antiinflammatory processes. In the present study we have shown that products of microfilarial (O. volvulus) and adult (O. volvulus and O. ochengi) parasites affect monocytes in vitro. An early production of TNF-alpha by exposed monocytes was followed by the production of IL-10 and a reduced expression of HLA-DR and the costimulatory molecules B7-1 and B7-2, while other adhesion receptors remained unaffected. Downregulation of the functional membrane receptors failed to occur after treatment of the cells with anti-IL-10 antibodies. The engagement of CD14, a dominant membrane receptor on monocytes and major binding protein for lipopolysaccharides, was indicated by partial blocking of monocyte modulation by neutralizing antibodies to CD14 and by the antagonistic lipid A analog compound 406. Lipopolysaccharide-like molecules were detected in sterile products of O. volvulus stages which could originate from Wolbachia bacteria related to Gram-negative Rickettsiales, known to be abundant in the hypodermis and the female reproductive organs of O. volvulus. The present results indicate that the monocyte/macrophage may be a major target cell for immunomodulatory parasite-derived and intraparasitic, bacteria-derived molecules, thereby contributing to the host's cellular hyporesponsiveness.}, } @article {pmid11007323, year = {2000}, author = {Lachaise, D and Harry, M and Solignac, M and Lemeunier, F and Bénassi, V and Cariou, ML}, title = {Evolutionary novelties in islands: Drosophila santomea, a new melanogaster sister species from São Tomé.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1452}, pages = {1487-1495}, pmid = {11007323}, issn = {0962-8452}, mesh = {Animals ; Atlantic Islands ; Bacterial Proteins/genetics ; *Biological Evolution ; Chimera ; Cytochrome c Group/genetics ; Drosophila/anatomy & histology/*classification/genetics/microbiology ; Drosophila melanogaster/anatomy & histology/classification/genetics/microbiology ; Female ; Genes, Insect ; Geography ; Male ; Wolbachia/genetics ; }, abstract = {The finding of new melanogaster sister species may help us in understanding more about how the emergence of genetic novelties, particularly in insular habitats, can result in speciation. Here we report on the discovery of Drosophila santomea, which is the first melanogaster sibling found off West-equatorial Africa, on São Tomé, one of the Gulf of Guinea islands. Although the eight other melanogaster sister species are remarkably conservative in their morphology except for their terminalia, the new find has a morphological trait distinguishing it from all of these: a pure yellow body coloration of both sexes without the normal black abdominal banding. Evidence from the terminalia, polytene and mitotic chromosomes, period gene and allozymes are provided indicating that it is nonetheless the nearest relative of Drosophila yakuba with which it coexists on the island. The new find is a clear-cut taxon as shown by the production of sterile male hybrids, eventually with developmental defects, in both directions of cross with yakuba and by the existence of an altitudinal divide accompanied by a hybrid zone at mid-elevation on the island. Molecular and karyotypic data further support this conclusion. In contrast to the significant divergence of their nuclear DNAs, an intriguing similarity in their cytochrome b sequences was observed indicating a recent coalescence common to santomea, yakuba and also teissieri cytoplasms. These were shown to harbour the same Wolbachia endosymbiotic bacteria which could possibly be responsible for mitochondrial DNA hitchhiking across the species barrier.}, } @article {pmid10983833, year = {2000}, author = {Fialho, RF and Stevens, L}, title = {Male-killing Wolbachia in a flour beetle.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1451}, pages = {1469-1473}, pmid = {10983833}, issn = {0962-8452}, mesh = {Animals ; Crosses, Genetic ; DNA/analysis ; Female ; Fertility ; Male ; Sequence Analysis, DNA ; Sex Ratio ; Tribolium/*microbiology ; Wolbachia/*physiology ; }, abstract = {The bacteria in the genus Wolbachia are cytoplasmically inherited symbionts of arthropods. Infection often causes profound changes in host reproduction, enhancing bacterial transmission and spread in a population. The reproductive alterations known to result from Wolbachia infection include cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males, fecundity enhancement, male killing and, perhaps, lethality Here, we report male killing in a third insect, the black flour beetle Tribolium madens, based on highly female-biased sex ratios of progeny from females infected with Wolbachia. The bias is cytoplasmic in nature as shown by repeated backcrossing of infected females with males of a naturally uninfected strain. Infection also lowers the egg hatch rates significantly to approximately half of those observed for uninfected females. Treatment of the host with antibiotics eliminated infection, reverted the sex ratio to unbiased levels and increased the percentage hatch. Typically Wolbachia infection is transmitted from mother to progeny, regardless of the sex of the progeny; however, infected T. madens males are never found. Virgin females are sterile, suggesting that the sex-ratio distortion in T. madens results from embryonic male killing rather than parthenogenesis. Based on DNA sequence data, the male-killing strain of Wolbachia in T. madens was indistinguishable from the CI-inducing Wolbachia in Tribolium confusum, a closely related beetle. Our findings suggest that host symbiont interaction effects may play an important role in the induction of Wolbachia reproductive phenotypes.}, } @article {pmid10972121, year = {2000}, author = {Werren, JH and Windsor, DM}, title = {Wolbachia infection frequencies in insects: evidence of a global equilibrium?.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1450}, pages = {1277-1285}, pmid = {10972121}, issn = {0962-8452}, mesh = {Animals ; DNA, Bacterial/genetics ; Insecta/*microbiology ; North America ; Polymerase Chain Reaction ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; *Wolbachia/genetics/isolation & purification ; }, abstract = {Wolbachia are a group of cytoplasmically inherited bacteria that cause reproduction alterations in arthropods, including parthenogenesis, reproductive incompatibility, feminization of genetic males and male killing. Previous general surveys of insects in Panama and Britain found Wolbachia to be common, occurring in 16-22% of species. Here, using similar polymerase chain reaction methods, we report that 19.3% of a sample of temperate North American insects are infected with Wolbachia, a frequency strikingly similar to frequencies found in two other studies in widely separated locales. The results may indicate a widespread equilibrium of Wolbachia infection frequencies in insects whose maintenance remains to be explained. Alternatively, Wolbachia may be increasing in global insect communities. Within each of the three geographic regions surveyed, Hymenoptera are more frequently infected with A group Wolbachia and Lepidoptera more frequently infected with B group Wolbachia.}, } @article {pmid10971717, year = {2000}, author = {Jeyaprakash, A and Hoy, MA}, title = {Long PCR improves Wolbachia DNA amplification: wsp sequences found in 76% of sixty-three arthropod species.}, journal = {Insect molecular biology}, volume = {9}, number = {4}, pages = {393-405}, doi = {10.1046/j.1365-2583.2000.00203.x}, pmid = {10971717}, issn = {0962-1075}, mesh = {Animals ; Arthropods/*microbiology ; Base Sequence ; *DNA, Bacterial ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods/standards ; Sensitivity and Specificity ; Wasps/*microbiology ; Wolbachia/classification/*genetics ; }, abstract = {Bacteria belonging to the genus Wolbachia are associated with a variety of reproductive anomalies in arthropods. Allele-specific polymerase chain reaction (= Standard PCR) routinely has been used to amplify Wolbachia DNA from arthropods. While testing the two-spotted spider mite Tetranychus urticae and other arthropods known to be infected with Wolbachia, Standard PCR frequently produced false negatives, perhaps because the DNA from the arthropod host interfered with amplification by Taq DNA polymerase. Long PCR, which uses two enzymes (Taq and Pwo), consistently amplified Wolbachia DNA and a sensitivity analysis indicated that Long PCR was approximately six orders of magnitude more sensitive than Standard PCR in amplifying plasmid DNA spiked into insect genomic DNA. A survey indicated that 76% of sixty-two arthropod species and two subspecies in thirteen orders tested positive for the Wolbachia wsp sequence by Long PCR, which is considerably higher than the rate of 16.9% obtained previously for the ftsZ sequence using Standard PCR (Werren, J.H., Windsor, D. and Gao, L. (1995a) Proc R Soc Lond B 262: 197-204). A subsample of Long PCR products from fourteen arthropod species and two subspecies were sequenced, both directly and after cloning. Two A- and eleven B-Wolbachia strains were detected and their wsp sequences displayed a maximum of 23.7% sequence divergence at this locus. Two new groups (named Fus and Ten) were identified in addition to nineteen reported earlier (Zhou, W., Rousset, F. and O'Neill, S.L. (1998) Proc R Soc Lond B 265: 1-7; van Meer, M.M.M., Witteveldt, J. and Stouthamer, R. (1999) Insect Mol Biol 8: 399-408), because they displayed more than 2.5% sequence divergence from other Wolbachia wsp sequences. PCR products from seventeen of twenty-nine (59%) arthropod species analysed could not be sequenced directly due to apparent infection by multiple Wolbachia strains. The wsp sequences cloned from two such species (Plutella xylostella and Trichoplusia ni) indicated both A- and B-Wolbachia were present in a single individual. Hence, superinfection also may be more widespread than the 1.2% incidence previously estimated.}, } @article {pmid10963399, year = {2000}, author = {Pintureau, B and Grenier, S and Boléat, B and Lassablière, F and Heddi, A and Khatchadourian, C}, title = {Dynamics of Wolbachia populations in transfected lines of Trichogramma.}, journal = {Journal of invertebrate pathology}, volume = {76}, number = {1}, pages = {20-25}, doi = {10.1006/jipa.2000.4953}, pmid = {10963399}, issn = {0022-2011}, mesh = {Animals ; Female ; Genetic Variation ; Microinjections ; Ovary/microbiology ; Solutions ; Symbiosis ; Wasps/growth & development/*microbiology ; Wolbachia/genetics/*growth & development/isolation & purification ; }, abstract = {Fluorescence in situ hybridization was tested to specifically detect symbionts of the genus Wolbachia in Trichogramma and to allow for semiquantitative estimations of symbiont abundance. Extraction solutions used for horizontal transfers of symbionts contain a high abundance of Wolbachia, but Wolbachia have a low and decreasing abundance in microinjected lines (transfected lines). Moreover, eggs of microinjected lines were shown to be polymorphic for the infection. In naturally infected lines, Wolbachia are localized at the posterior pole of the eggs; they are scattered during the early stages of larval development and then concentrated in the ovaries at the end of the female pupal development. Scattering and concentration are probably not active but rather the result of replications or morphogenesis. Conversely, Wolbachia are not concentrated at the posterior pole of eggs in microinjected lines. Comparison of the within-family and between-family variances of the symbiont abundance in a microinjected line did not lead us to conclude that this character shows a genetic variability.}, } @article {pmid10948369, year = {2000}, author = {De Barro, PJ and Hart, PJ}, title = {Mating interactions between two biotypes of the whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) in Australia.}, journal = {Bulletin of entomological research}, volume = {90}, number = {2}, pages = {103-112}, doi = {10.1017/s0007485300000201}, pmid = {10948369}, issn = {0007-4853}, mesh = {Animals ; Australia ; Female ; Fertility ; Hemiptera/*physiology ; Male ; Sexual Behavior, Animal/*physiology ; }, abstract = {The biological consequences of mating interactions between indigenous and exotic biotypes of Bemisia tabaci (Gennadius) in Australia were studied using a combination of field and laboratory experiments. The key results of the interaction between the B and eastern Australian biotypes were reduced population increase, a marked increase in the proportion of male progeny, fewer eggs produced by females paired with males of different biotype and no difference in the numbers of eggs per unmated female and females paired with males of the same biotype. In addition, there was no change in the proportion of eggs hatching, mixed biotype pairs spent more time courting than single biotype pairs and a low level of hybridization in field cages and small containers was observed. These observations suggest three possibilities. The first is the 'distracting male hypothesis' in which mating pairs made up of different biotypes apportion more time to courtship and less time to egg laying than single biotype pairs. The second invokes the 'single-locus complementary sex determination model' in which the production of non-viable diploid male zygotes may explain the reduction in eggs laid. The third is cytoplasmic incompatibility between biotypes caused by Wolbachia. The results also suggest that the geographical distribution of clusters of related biotypes both overseas and in Australia may be explained by between-biotype interactions leading to the formation of parapatric populations.}, } @article {pmid10937195, year = {2000}, author = {Vavre, F and Fleury, F and Varaldi, J and Fouillet, P and Boulétreau, M}, title = {Evidence for female mortality in Wolbachia-mediated cytoplasmic incompatibility in haplodiploid insects: epidemiologic and evolutionary consequences.}, journal = {Evolution; international journal of organic evolution}, volume = {54}, number = {1}, pages = {191-200}, doi = {10.1111/j.0014-3820.2000.tb00019.x}, pmid = {10937195}, issn = {0014-3820}, mesh = {Animals ; Base Sequence ; *Biological Evolution ; Cytoplasm/genetics ; DNA Primers/genetics ; Diploidy ; Drosophila/genetics/microbiology ; Female ; Haploidy ; Hymenoptera/genetics/microbiology ; Insecta/*genetics/*microbiology ; Male ; Parthenogenesis ; Phylogeny ; Reproduction ; Wolbachia/genetics/*pathogenicity ; }, abstract = {Until now, only two Wolbachia-mediated cytoplasmic incompatibility (CI) types have been described in haplodiploid species, the first in Nasonia (Insect) and the second in Tetranychus (Acari). They both induce a male-biased sex ratio in the incompatible cross. In Nasonia, CI does not reduce fertility since incompatible eggs develop as haploid males, whereas in Tetranychus CI leads to a partial mortality of incompatible eggs, thus reducing the fertility of females. Here, we study Wolbachia infection in a Drosophila parasitoid, Leptopilina heterotoma (Hymenoptera: Figitidae). A survey of Wolbachia infection shows that all natural populations tested are totally infected. Crosses between infected males and cured females show complete incompatibility: almost no females are produced. Moreover, incompatible eggs die early during their development, unlike Nasonia. This early death allows the parasitized Drosophila larva to achieve its development and to emerge. Thus, uninfected females crossed with infected males have reduced offspring production consisting only of males. Evidence of this CI type in insects demonstrates that the difference in CI types of Nasonia and Tetranychus is not due to specific factors of insects or acari. Using theoretical models, we compare the invasion processes of different strategies of Wolbachia: CI in diploid species, the two CI types in haplodiploid species, and parthenogenesis (the classical effect in haplodiploid species). Models show that CI in haplodiploid species is less efficient than in diploid ones. However, the Leptopilina type is advantageous compared to the Nasonia type. Parthenogenesis may be more or less advantageous, depending on the infection cost and on the proportion of fertilized eggs. Finally, we can propose different processes of Wolbachia strategy evolution in haplodiploid species from Nasonia CI type to Leptopilina CI type or parthenogenesis.}, } @article {pmid10903373, year = {2000}, author = {Ballard, JW}, title = {Comparative genomics of mitochondrial DNA in Drosophila simulans.}, journal = {Journal of molecular evolution}, volume = {51}, number = {1}, pages = {64-75}, doi = {10.1007/s002390010067}, pmid = {10903373}, issn = {0022-2844}, mesh = {Alcohol Dehydrogenase/genetics ; Animals ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics ; *Evolution, Molecular ; Genetic Variation ; Genome ; Haplotypes ; Models, Genetic ; *Phylogeny ; Polymorphism, Genetic ; }, abstract = {The current study compares the nucleotide variation among 22 complete mitochondrial genomes of the three distinct Drosophila simulans haplotypes with intron 1 of the alcohol dehydrogenase-related locus. This is the first study to investigate the sequence variation of multiple complete mitochondrial genomes within distinct mitochondrial haplotypes of a single species. Patterns of variation suggest distinct forces are influencing the evolution of mitochondrial DNA (mtDNA) and autosomal DNA in D. simulans. First, there is little variation within each mtDNA haplotype but strong differentiation among them. In contrast, there is no support for differentiation of the mitochondrial haplotypes at the autosomal locus. Second, there is a significant deficiency of mitochondrial variation in each haplotype relative to the autosomal locus. Third, the ratio of nonsynonymous to synonymous substitutions is not equal in all branches of the well-resolved phylogeny. There is an excess of nonsynonymous substitutions relative to synonymous substitutions within each D. simulans haplotype. This result is similar to that previously observed within the mtDNA of distinct species. A single evolutionary force may be causally linked to the observed patterns of mtDNA variation-a rickettsia-like microorganism, Wolbachia pipientis, which is known to directly influence mitochondrial evolution but have a less direct influence on autosomal loci.}, } @article {pmid10903165, year = {2000}, author = {Kamoda, S and Masui, S and Ishikawa, H and Sasaki, T}, title = {Wolbachia infection and cytoplasmic incompatibility in the cricket Teleogryllus taiwanemma.}, journal = {The Journal of experimental biology}, volume = {203}, number = {Pt 16}, pages = {2503-2509}, doi = {10.1242/jeb.203.16.2503}, pmid = {10903165}, issn = {0022-0949}, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Cytoplasm/microbiology ; Female ; Genes, Bacterial ; Gryllidae/*microbiology/*physiology ; Male ; Phylogeny ; Proteins/isolation & purification ; Reproduction ; Spermatozoa/chemistry ; Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are cytoplasmically inherited bacteria found in many arthropods. They induce various reproductive alterations in their hosts, including cytoplasmic incompatibility, thelytokous parthenogenesis, feminization and male-killing. In this study, we examined Wolbachia infection and its effects on the host cricket Teleogryllus taiwanemma. In a phylogenetic study based on the wsp gene coding for a Wolbachia surface protein, the Wolbachia strain harboured by T. taiwanemma was clustered together with those harboured by Laodelphax striatellus, Tribolium confusum, Acraea encedon, Trichogramma deion and Adalia bipunctata. Crossing experiments using the Wolbachia-infected and uninfected strains of cricket showed that the infection is associated with the expression of unidirectional cytoplasmic incompatibility: the egg hatch rate in the incompatible cross between the infected males and uninfected females was 20.3 %. We also examined the distribution of Wolbachia within the host using polymerase chain reaction assays; they were detected in the antennae, heads, forewings, hindwings, testes, ovaries, Malpighian tubules, foot muscles and fat bodies. Quantitative polymerase chain reaction assays showed that the bacterial density was highest in the fat bodies, followed by the ovaries and testes. Wolbachia were not detected in the haemolymph or in mature spermatozoa. The spermatozoa of the infected male may be modified by the presence of Wolbachia during its development. To examine this possibility, we compared the profiles of sperm proteins between the infected and uninfected males using two-dimensional gel electrophoresis. However, no differences in the protein profiles were observed.}, } @article {pmid10885510, year = {2000}, author = {Langworthy, NG and Renz, A and Mackenstedt, U and Henkle-Dührsen, K and de Bronsvoort, MB and Tanya, VN and Donnelly, MJ and Trees, AJ}, title = {Macrofilaricidal activity of tetracycline against the filarial nematode Onchocerca ochengi: elimination of Wolbachia precedes worm death and suggests a dependent relationship.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1448}, pages = {1063-1069}, pmid = {10885510}, issn = {0962-8452}, mesh = {Animals ; Cameroon ; Cattle ; Female ; Humans ; Male ; Onchocerca/cytology/*drug effects/*microbiology ; Onchocerciasis/*drug therapy ; Onchocerciasis, Ocular/parasitology ; Oxytetracycline/*pharmacology/therapeutic use ; Tetracyclines/*pharmacology/therapeutic use ; Wolbachia/drug effects/*physiology ; }, abstract = {Filarial nematodes are important and widespread parasites of animals and humans. We have been using the African bovine parasite Onchocerca ochengi as a chemotherapeutic model for O. volvulus, the causal organism of 'river blindness' in humans, for which there is no safe and effective drug lethal to adult worms. Here we report that the antibiotic, oxytetracycline is macrofilaricidal against O. ochengi. In a controlled trial in Cameroon, all adult worms (as well as microfilariae) were killed, and O. ochengi intradermal nodules resolved, by nine months' post-treatment in cattle treated intermittently for six months. Adult worms removed from concurrent controls remained fully viable and reproductively active. By serial electron-microscopic examination, the macrofilaricidal effects were related to the elimination of intracellular micro-organisms, initially abundant. Analysis of a fragment of the 16S rRNA gene from the O. ochengi micro-organisms confirmed them to be Wolbachia organisms of the order Rickettsiales, and showed that the sequence differed in only one nucleotide in 858 from the homologous sequence of the Wolbachia organisms of O. volvulus. These data are, to our knowledge, the first to show that antibiotic therapy can be lethal to adult filariae. They suggest that tetracycline therapy is likely to be macrofilaricidal against O. volvulus infections in humans and, since similar Wolbachia organisms occur in a number of other filarial nematodes, against those infections too. In that the elimination of Wolbachia preceded the resolution of the filarial infections, they suggest that in O. ochengi at least, the Wolbachia organisms play an essential role in the biology and metabolism of the filarial worm.}, } @article {pmid10874716, year = {2000}, author = {Trees, AJ and Graham, SP and Renz, A and Bianco, AE and Tanya, V}, title = {Onchocerca ochengi infections in cattle as a model for human onchocerciasis: recent developments.}, journal = {Parasitology}, volume = {120 Suppl}, number = {}, pages = {S133-42}, doi = {10.1017/s0031182099005788}, pmid = {10874716}, issn = {0031-1820}, mesh = {Animals ; Cattle ; *Cattle Diseases/drug therapy/immunology ; Disease Models, Animal ; Female ; Host-Parasite Interactions ; Humans ; Male ; *Onchocerca/genetics/immunology ; Onchocerciasis/*veterinary ; }, abstract = {The bovine parasite Onchocerca ochengi is a nodule-dwelling filarial nematode, closely related to O. volvulus, the causal agent of human River Blindness, and, sharing with it, the same vector. This brief review, based on a presentation at the BSP Autumn Symposium 1999, describes recent work supported by the WHO Drug Development Research Macrofil programme and the Edna McConnell Clark Foundation vaccine development programme, to research the chemotherapy and immunology of onchocerciasis utilising this model system, with experimental infections in Liverpool and field infections in northern Cameroon. In a series of chemotherapeutic trials involving 10 compounds in 20 treatment regimes, the comparability of drug efficacy against O. ochengi with that described against O. volvulus has been demonstrated. Repeated, long-term treatment with oxytetracycline has been shown to be macrofilaricidal and the effect is hypothesized to be related to action on Wolbachia endobacteria, abundant in O. ochengi. Avermectins/milbemycins are not macrofilaricidal (even in high and repeated long-term treatments) but induce sustained abrogation of embryogenesis. In prospective, field exposure experiments with naive calves, prophylactic treatments with ivermectin and moxidectin prevented the development of adult worm infection, raising the possibility that drug-attenuated larval challenge infections may induce immunity. Putatively immune adult cattle exist in endemically exposed populations, and these have been shown to be significantly less susceptible to challenge than age-matched naive controls, whereas radically drug-cured, previously patently-infected cattle were not. Experimental infections with O. ochengi have revealed the kinetics of the immune response in relation to parasite development and demonstrate analogous responses to those reported in O. volvulus infection in humans and chimpanzees. In an immunization experiment with irradiated L3 larvae, cattle were significantly protected against experimental challenge--the first such demonstration of the experimental induction of immunity in a natural Onchocerca host-parasite system. Taken collectively, these studies not only demonstrate the similarity between the host-parasite relationships of O. ochengi in cattle and O. volvulus in humans, but promise to advance options for the control of human onchocerciasis.}, } @article {pmid10856373, year = {2000}, author = {Bazzocchi, C and Jamnongluk, W and O'Neill, SL and Anderson, TJ and Genchi, C and Bandi, C}, title = {wsp gene sequences from the Wolbachia of filarial nematodes.}, journal = {Current microbiology}, volume = {41}, number = {2}, pages = {96-100}, doi = {10.1007/s002840010100}, pmid = {10856373}, issn = {0343-8651}, support = {AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Brugia malayi/microbiology ; Cloning, Molecular ; DNA Primers ; Dirofilaria/microbiology ; *Genes, Bacterial ; Molecular Sequence Data ; Nematoda/*microbiology ; Phylogeny ; Symbiosis/genetics ; Wolbachia/chemistry/*genetics ; Wuchereria bancrofti/microbiology ; }, abstract = {Wolbachia endosymbiotic bacteria are widespread in arthropods and are also present in filarial nematodes. Almost all filarial species so far examined have been found to harbor these endosymbionts. The sequences of only three genes have been published for nematode Wolbachia (i.e., the genes coding for the proteins FtsZ and catalase and for 16S rRNA). Here we present the sequences of the genes coding for the Wolbachia surface protein (WSP) from the endosymbionts of eight species of filaria. Complete gene sequences were obtained from the endosymbionts of two different species, Dirofilaria immitis and Brugia malayi. These sequences allowed us to design general primers for amplification of the wsp gene from the Wolbachia of all filarial species examined. For these species, partial WSP sequences (about 600 base pairs) were obtained with these primers. Phylogenetic analysis groups these nematode wsp sequences into a coherent cluster. Within the nematode cluster, wsp-based Wolbachia phylogeny matches a previous phylogeny obtained with ftsZ gene sequences, with a good consistency of the phylogeny of hosts (nematodes) and symbionts (Wolbachia). In addition, different individuals of the same host species (Dirofilaria immitis and Wuchereria bancrofti) show identical wsp gene sequences.}, } @article {pmid10853728, year = {2000}, author = {Randerson, JP and Jiggins, FM and Hurst, LD}, title = {Male killing can select for male mate choice: a novel solution to the paradox of the lek.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1446}, pages = {867-874}, pmid = {10853728}, issn = {0962-8452}, mesh = {Animals ; Butterflies/*genetics/microbiology/*physiology ; Female ; Genes, Insect ; Genetic Variation ; Male ; Models, Biological ; Sex Ratio ; Sexual Behavior, Animal ; Wolbachia/pathogenicity ; }, abstract = {In lekking species, intense directional selection is applied to aspects of the male genotype by female choice. Under conventional quantitative genetics theory, the expectation is that this will lead to a rapid loss in additive genetic variance for the trait in question. However, despite female choice, male variation is maintained and hence it pays females to continue choosing. This has been termed the 'paradox of the lek'. Here we present a theoretical analysis of a putative sex-role-reversed lek in the butterfly Acraea encedon. Sex-role reversal appears to have come about because of infection with a male-killing Wolbachia. The bacterium is highly prevalent in some populations, such that there is a dearth of males. Receptive females form dense aggregations, and it has been suggested that males preferentially select females uninfected with the bacterium. As with more conventional systems, this presents a theoretical problem exactly analogous to the lek paradox, namely what maintains female variation and hence why do males continue to choose? We model the evolution of a male choice gene that allows discrimination between infected and uninfected females, and show that the stable maintenance of both female variation and male choice is likely, so long as males make mistakes when discriminating between females. Furthermore, our model allows the maintenance, in a panmictic population, of a male killer that is perfectly transmitted. This is the first model to allow this result, and may explain the long-term persistence of a male killer in Hypolimnas bolina.}, } @article {pmid10843833, year = {2000}, author = {Zchori-Fein, E and Gottlieb, Y and Coll, M}, title = {Wolbachia density and host fitness components in Muscidifurax uniraptor (Hymenoptera: pteromalidae).}, journal = {Journal of invertebrate pathology}, volume = {75}, number = {4}, pages = {267-272}, doi = {10.1006/jipa.2000.4927}, pmid = {10843833}, issn = {0022-2011}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Colony Count, Microbial ; Hymenoptera/drug effects/*microbiology ; Reproduction/drug effects ; Rifampin/pharmacology ; Sex Ratio ; Wolbachia/drug effects/*isolation & purification ; }, abstract = {Intracellular bacteria of the genus Wolbachia are found in a variety of arthropod hosts, where they cause various reproductive disorders. Attempts to study the fitness advantages and disadvantages of carrying these symbionts have yielded contradicting results. Using various doses of the antibiotic rifampicin, we were able to manipulate the density of Wolbachia in the uniparental parasitoid Muscidifurax uniraptor (Hymenoptera: Pteromalidae). The effect of different titers of the symbiont on the fecundity, reproductive rate, longevity, survival rate, and sex ratio of the host was measured. The data gathered show that following antibiotic treatments, the percentage of males rises at low doses of rifampicin and then drops again. The total sex ratio of offspring produced by treated mothers was positively correlated with the numbers of Wolbachia found in eggs laid by these females. No significant effects were detected with regard to the other studied fitness components. It is concluded that in M. uniraptor, Wolbachia are not posing any burden on the life history trait studied.}, } @article {pmid10849086, year = {2000}, author = {Majerus, ME and Hinrich, J and Schulenburg, GV and Zakharov, IA}, title = {Multiple causes of male-killing in a single sample of the two-spot ladybird, Adalia bipunctata (Coleoptera: coccinellidae) from Moscow.}, journal = {Heredity}, volume = {84 (Pt 5)}, number = {}, pages = {605-609}, doi = {10.1046/j.1365-2540.2000.00710.x}, pmid = {10849086}, issn = {0018-067X}, mesh = {Animals ; Coleoptera/genetics/*microbiology ; DNA Primers/chemistry ; Infectious Disease Transmission, Vertical ; Male ; Moscow ; Phenotype ; Polymerase Chain Reaction ; Rickettsieae/*physiology ; Sequence Analysis, DNA ; Sex Ratio ; Spiroplasma/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Thirty-six matrilines from a single Muscovite sample of Adalia bipunctata were assayed, using appropriate primers, for presence of the four male-killing symbionts known to infect this species of ladybird. All four, a Rickettsia, a Spiroplasma and two different strains of Wolbachia, were found to be present. Vertical transmission efficiencies were assessed from F1 and F2 families from each of the matrilines, and were found to differ significantly between symbionts. Potential explanations of the presence of four different male-killing symbionts within a single population, are considered.}, } @article {pmid10822808, year = {2000}, author = {Zakharov, IA and Goriacheva, II and Shaĭkevich, EV and Schulenburg, JH and Majerus, EN}, title = {[Wolbachia--a new bacteria causing sex ratio bias in the two-spot lady-bird Adalia bipunctata L].}, journal = {Genetika}, volume = {36}, number = {4}, pages = {482-486}, pmid = {10822808}, issn = {0016-6758}, mesh = {Animals ; Female ; Insecta/*microbiology/*physiology ; Male ; Sex Factors ; Sex Ratio ; Wolbachia/*isolation & purification ; }, abstract = {Some of the male-killing lines of the two-spot ladybird Adalia bipunctata L. isolated from the populations of Moscow and Tomsk and having a female-biased sex ratio were found to be infected with a bacterium of the genus Wolbachia. This fact is the first demonstration of the ability of Wolbachia to kill males of a host insect. The coexistence of females infected with different male-killing bacteria was recorded in the population of Moscow.}, } @article {pmid10821272, year = {2000}, author = {Huigens, ME and Luck, RF and Klaassen, RH and Maas, MF and Timmermans, MJ and Stouthamer, R}, title = {Infectious parthenogenesis.}, journal = {Nature}, volume = {405}, number = {6783}, pages = {178-179}, doi = {10.1038/35012066}, pmid = {10821272}, issn = {0028-0836}, mesh = {Animals ; Butterflies/parasitology ; Female ; Larva/microbiology ; Ovum/microbiology ; *Parthenogenesis ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {Parthenogenesis-inducing Wolbachia bacteria are reproductive parasites that cause infected female wasps to produce daughters without mating. This manipulation of the host's reproduction enhances the transmission of Wolbachia to future generations because the bacteria are passed on vertically only from mothers to daughters. Males are dead ends for cytoplasmically inherited bacteria: they do not pass them on to their offspring. Vertical transmission of Wolbachia has been previously considered to be the main mode of transmission. Here we report frequent horizontal transmission from infected to uninfected wasp larvae sharing a common food source. The transferred Wolbachia are then vertically transmitted to the new host's offspring. This natural and unexpectedly frequent horizontal transfer of parthenogensis-inducing Wolbachia intraspecifically has important implications for the co-evolution of Wolbachia and their host.}, } @article {pmid10810024, year = {2000}, author = {Hoerauf, A}, title = {Targeting of wolbachia endobacteria in litomosoides sigmodontis: comparison of tetracyclines with chloramphenicol, macrolides and ciprofloxacin.}, journal = {Tropical medicine & international health : TM & IH}, volume = {5}, number = {4}, pages = {275-279}, pmid = {10810024}, issn = {1365-3156}, abstract = {Endobacteria of the genus Wolbachia in filarial nematodes are related to Rickettsiaceae and can be depleted by tetracycline antibiotics. This depletion blocks female worm development as well as early embryogenesis, in contrast to the currently used microfilaricidal ivermectin which blocks only the last stage of embryogenesis. Since targeting Wolbachia is becoming an area of research for the treatment of human filariases, it was investigated if antibiotics other than tetracyclines are able to deplete Wolbachia from filariae. BALB/c mice infected with the rodent filaria Litomosoides sigmodontis were treated with erythromycin, chloramphenicol or ciprofloxacin. All drugs were well resorbed and resulted in serum levels clearly above breakpoint levels for bacteria susceptible to the respective antibiotic. However, contrary to tetracycline, none of these antibiotics depleted Wolbachia or altered worm development and fertility, as evidenced by immunohistology, immunoelectron microscopy and semiquantitative PCR.}, } @article {pmid10810023, year = {2000}, author = {Hoerauf, A and Volkmann, L and Nissen-Paehle, K and Schmetz, C and Autenrieth, I and Büttner, DW and Fleischer, B}, title = {Targeting of Wolbachia endobacteria in Litomosoides sigmodontis: comparison of tetracyclines with chloramphenicol, macrolides and ciprofloxacin.}, journal = {Tropical medicine & international health : TM & IH}, volume = {5}, number = {4}, pages = {275-279}, pmid = {10810023}, issn = {1360-2276}, mesh = {Animals ; Anti-Bacterial Agents/blood/*therapeutic use ; Chloramphenicol/blood/therapeutic use ; Ciprofloxacin/blood/therapeutic use ; DNA, Bacterial/*isolation & purification ; Female ; Filariasis/*drug therapy ; Filarioidea/*growth & development/microbiology ; Injections, Intraperitoneal ; Macrolides ; Mice ; Mice, Inbred BALB C ; Polymerase Chain Reaction ; Tetracycline/blood/therapeutic use ; Wolbachia/*drug effects ; }, abstract = {Endobacteria of the genus Wolbachia in filarial nematodes are related to Rickettsiaceae and can be depleted by tetracycline antibiotics. This depletion blocks female worm development as well as early embryogenesis, in contrast to the currently used microfilaricidal ivermectin which blocks only the last stage of embryogenesis. Since targeting Wolbachia is becoming an area of research for the treatment of human filariases, it was investigated if antibiotics other than tetracyclines are able to deplete Wolbachia from filariae. BALB/c mice infected with the rodent filaria Litomosoides sigmodontis were treated with erythromycin, chloramphenicol or ciprofloxacin. All drugs were well resorbed and resulted in serum levels clearly above breakpoint levels for bacteria susceptible to the respective antibiotic. However, contrary to tetracycline, none of these antibiotics depleted Wolbachia or altered worm development and fertility, as evidenced by immunohistology, immunoelectron microscopy and semiquantitative PCR.}, } @article {pmid10790392, year = {2000}, author = {Snook, RR and Cleland, SY and Wolfner, MF and Karr, TL}, title = {Offsetting effects of Wolbachia infection and heat shock on sperm production in Drosophila simulans: analyses of fecundity, fertility and accessory gland proteins.}, journal = {Genetics}, volume = {155}, number = {1}, pages = {167-178}, pmid = {10790392}, issn = {0016-6731}, support = {1T34GM08491/GM/NIGMS NIH HHS/United States ; HD08172-01/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*microbiology/*physiology ; *Drosophila Proteins ; Female ; Fertility/physiology ; Heat-Shock Response/physiology ; Intercellular Signaling Peptides and Proteins ; Male ; Peptides/*metabolism ; Protein Processing, Post-Translational ; Sexual Behavior, Animal ; Spermatozoa/*cytology/microbiology/physiology ; Symbiosis/physiology ; Wolbachia/*physiology ; }, abstract = {Infection in Drosophila simulans with the endocellular symbiont Wolbachia pipientis results in egg lethality caused by failure to properly initiate diploid development (cytoplasmic incompatibility, CI). The relationship between Wolbachia infection and reproductive factors influencing male fitness has not been well examined. Here we compare infected and uninfected strains of D. simulans for (1) sperm production, (2) male fertility, and (3) the transfer and processing of two accessory gland proteins, Acp26Aa or Acp36De. Infected males produced significantly fewer sperm cysts than uninfected males over the first 10 days of adult life, and infected males, under varied mating conditions, had lower fertility compared to uninfected males. This fertility effect was due to neither differences between infected and uninfected males in the transfer and subsequent processing of accessory gland proteins by females nor to the presence of Wolbachia in mature sperm. We found that heat shock, which is known to decrease CI expression, increases sperm production to a greater extent in infected compared to uninfected males, suggesting a possible link between sperm production and heat shock. Given these results, the roles Wolbachia and heat shock play in mediating male gamete production may be important parameters for understanding the dynamics of infection in natural populations.}, } @article {pmid10782070, year = {2000}, author = {Taylor, MJ and Bandi, C and Hoerauf, AM and Lazdins, J}, title = {Wolbachia bacteria of filarial nematodes: a target for control?.}, journal = {Parasitology today (Personal ed.)}, volume = {16}, number = {5}, pages = {179-180}, doi = {10.1016/s0169-4758(00)01661-6}, pmid = {10782070}, issn = {0169-4758}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology/*therapeutic use ; Brugia malayi/drug effects/microbiology ; Filariasis/*drug therapy/microbiology/parasitology ; Filarioidea/drug effects/*microbiology ; Humans ; Onchocerca volvulus/drug effects/microbiology ; Rifampin/therapeutic use ; Symbiosis ; Tetracycline/pharmacology/*therapeutic use ; Wolbachia/drug effects/*pathogenicity ; Wuchereria bancrofti/drug effects/microbiology ; }, } @article {pmid10770808, year = {2000}, author = {Taylor, MJ and Cross, HF and Bilo, K}, title = {Inflammatory responses induced by the filarial nematode Brugia malayi are mediated by lipopolysaccharide-like activity from endosymbiotic Wolbachia bacteria.}, journal = {The Journal of experimental medicine}, volume = {191}, number = {8}, pages = {1429-1436}, pmid = {10770808}, issn = {0022-1007}, mesh = {Animals ; Brugia malayi/*microbiology/*pathogenicity ; Cell Line ; Culicidae ; Filariasis/*etiology ; Humans ; Inflammation/*etiology ; Inflammation Mediators/isolation & purification/toxicity ; Lipopolysaccharides/isolation & purification/toxicity ; Mice ; Mice, Inbred C3H ; Symbiosis ; Wolbachia/*pathogenicity ; }, abstract = {The pathogenesis of filarial disease is characterized by acute and chronic inflammation. Inflammatory responses are thought to be generated by either the parasite, the immune response, or opportunistic infection. We show that soluble extracts of the human filarial parasite Brugia malayi can induce potent inflammatory responses, including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and nitric oxide (NO) from macrophages. The active component is heat stable, reacts positively in the Limulus amebocyte lysate assay, and can be inhibited by polymyxin B. TNF-alpha, IL-1beta, and NO responses were not induced in macrophages from lipopolysaccharide (LPS)-nonresponsive C3H/HeJ mice. The production of TNF-alpha after chemotherapy of microfilariae was also only detected in LPS-responsive C3H/HeN mice, suggesting that signaling through the Toll-like receptor 4 (TLR4) is necessary for these responses. We also show that CD14 is required for optimal TNF-alpha responses at low concentrations. Together, these results suggest that extracts of B. malayi contain bacterial LPS. Extracts from the rodent filaria, Acanthocheilonema viteae, which is not infected with the endosymbiotic Wolbachia bacteria found in the majority of filarial parasites, failed to induce any inflammatory responses from macrophages, suggesting that the source of bacterial LPS in extracts of B. malayi is the Wolbachia endosymbiont. Wolbachia extracts derived from a mosquito cell line induced similar LPS-dependent TNF-alpha and NO responses from C3H/HeN macrophages, which were eliminated after tetracycline treatment of the bacteria. Thus, Wolbachia LPS may be one of the major mediators of inflammatory pathogenesis in filarial nematode disease.}, } @article {pmid10770311, year = {2000}, author = {Hoerauf, A and Volkmann, L and Hamelmann, C and Adjei, O and Autenrieth, IB and Fleischer, B and Büttner, DW}, title = {Endosymbiotic bacteria in worms as targets for a novel chemotherapy in filariasis.}, journal = {Lancet (London, England)}, volume = {355}, number = {9211}, pages = {1242-1243}, doi = {10.1016/S0140-6736(00)02095-X}, pmid = {10770311}, issn = {0140-6736}, mesh = {Adolescent ; Adult ; Animals ; Anti-Bacterial Agents/pharmacology/*therapeutic use ; DNA, Bacterial/analysis ; DNA, Helminth/analysis ; Doxycycline/pharmacology/*therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Onchocerca volvulus/genetics/*microbiology ; Onchocerciasis/*drug therapy/microbiology ; Polymerase Chain Reaction ; Symbiosis ; Wolbachia/*drug effects/genetics/physiology ; }, abstract = {Endosymbiotic bacteria living in plasmodia or worm parasites are required for the homoeostasis of their host and should be excellent targets for chemotherapy of certain parasitic diseases. We show that targeting of Wolbachia spp bacteria in Onchocerca volvulus filariae by doxycycline leads to sterility of adult worms to an extent not seen with drugs used against onchocerciasis, a leading cause of blindness in African countries.}, } @article {pmid10759311, year = {2000}, author = {Cheng, Q and Ruel, TD and Zhou, W and Moloo, SK and Majiwa, P and O'Neill, SL and Aksoy, S}, title = {Tissue distribution and prevalence of Wolbachia infections in tsetse flies, Glossina spp.}, journal = {Medical and veterinary entomology}, volume = {14}, number = {1}, pages = {44-50}, doi = {10.1046/j.1365-2915.2000.00202.x}, pmid = {10759311}, issn = {0269-283X}, support = {AI-34033/AI/NIAID NIH HHS/United States ; AI-40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Insect Control ; Insect Vectors ; Polymerase Chain Reaction/veterinary ; Tsetse Flies/*microbiology ; *Wolbachia ; }, abstract = {Tsetse flies Glossina spp. (Diptera: Glossinidae) harbor three different symbiotic microorganisms, one being an intracellular Rickettsia of the genus Wolbachia. This bacterium infects a wide range of arthropods, where it causes a variety of reproductive abnormalities, one of which is termed cytoplasmic incompatibility (CI) that, when expressed, results in embryonic death due to disruptions in fertilization events. We report here that in colonized flies, Wolbachia infections can be detected in 100% of sampled individuals, while infections vary significantly in field populations. Based on Wolbachia Surface Protein (wsp) gene sequence analysis, the infections associated with different fly species are all unique within the A group of the Wolbachia pipientis clade. In addition to being present in germ-line tissues, Wolbachia infections have been found in somatic tissues of several insects. Using a Wolbachia-specific PCR-based assay, the tissue tropism of infections in Glossina morsitans morsitans Westwood, Glossina brevipalpis Newstead and Glossina austeni Newstead were analysed. While infections in G. m. morsitans and G. brevipalpis were limited to reproductive tissues, in G. austeni, Wolbachia could be detected in various somatic tissues.}, } @article {pmid10742050, year = {2000}, author = {Schulenburg, JH and Hurst, GD and Huigens, TM and van Meer, MM and Jiggins, FM and Majerus, ME}, title = {Molecular evolution and phylogenetic utility of Wolbachia ftsZ and wsp gene sequences with special reference to the origin of male-killing.}, journal = {Molecular biology and evolution}, volume = {17}, number = {4}, pages = {584-600}, doi = {10.1093/oxfordjournals.molbev.a026338}, pmid = {10742050}, issn = {0737-4038}, mesh = {Bacterial Outer Membrane Proteins/*genetics ; Bacterial Proteins/*genetics ; *Cytoskeletal Proteins ; Evolution, Molecular ; *Phylogeny ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {A detailed assessment of the evolution and phylogenetic utility of two genes, ftsZ and wsp, was used to investigate the origin of male-killing Wolbachia, previously isolated from the ladybird Adalia bipunctata and the butterfly Acraea encedon. The analysis included almost all available sequences of B-group Wolbachia and two outgroup taxa and showed that (1) the two gene regions differ in phylogenetic utility, (2) sequence variation is here correlated with phylogenetic information content, (3) both genes show significant rate heterogeneity between lineages, (4) increased substitution rates are associated with homoplasy in the data, (5) wsp sequences of some taxa appear to be subject to positive selection, and (6) only a limited number of clades can be inferred with confidence due to either lack of phylogenetic information or the presence of homoplasy. With respect to the evolution of male-killing, the two genes nevertheless seemed to provide unbiased information. However, they consistently produce contradictory results. Current data therefore do not permit clarification of the origin of this behavior. In addition, A. bipunctata was found to be a host to two recently diverged strains of male-killing Wolbachia that showed increased substitution rates for both genes. Moreover, the wsp gene, which codes for an outer membrane protein, was found to be subject to positive selection in these taxa. These findings were postulated to be the product of high selection pressures due to antagonistic host-symbiont interactions in this ladybird species. In conclusion, our study demonstrates that the results of a detailed phylogenetic analysis, including characterization of the limitations of such an approach, can serve as a valuable basis for an understanding of the evolution of Wolbachia bacteria. Moreover, particular features of gene evolution, such as elevated substitution rates or the presence of positive selection, may provide information about the dynamics of Wolbachia-host associations.}, } @article {pmid10731575, year = {2000}, author = {Unnasch, TR and Williams, SA}, title = {The genomes of Onchocerca volvulus.}, journal = {International journal for parasitology}, volume = {30}, number = {4}, pages = {543-552}, doi = {10.1016/s0020-7519(99)00184-8}, pmid = {10731575}, issn = {0020-7519}, mesh = {Animals ; DNA, Mitochondrial/chemistry ; Gene Expression ; Genetic Variation ; *Genome ; Onchocerca volvulus/*genetics ; }, abstract = {Onchocerca volvulus, the filarial parasite that causes onchocerciasis or river blindness, contains three distinct genomes. These include the nuclear genome, the mitochondrial genome and the genome of an intracellular endosymbiont of the genus Wolbachia. The nuclear genome is roughly 1.5x10(8) bp in size, and is arranged on four chromosome pairs. Analysis of expressed sequence tags from different life-cycle stages has resulted in the identification of transcripts from roughly 4000 O. volvulus genes. Several of these transcripts are highly abundant, including those encoding collagen and cuticular proteins. Analysis of several gene sequences from O. volvulus suggests that the nuclear genes of O. volvulus are relatively compact and are interrupted relatively frequently by small introns. The intron-exon boundaries of these genes generally follow the GU-AG rule characteristic of the splice donor and acceptors of other vertebrate organisms. The nuclear genome also contains at least one repeated sequence family of a 150 bp repeat which is arranged in tandem arrays and appears subject to concerted evolution. The mitochondrial genome of O. volvulus is remarkably compact, only 13747 bp in size. Consistent with the small size of the genome, four gene pairs overlap, eight contain no intergenic regions and the remaining gene pairs are separated by small intergenic domains ranging from 1 to 46 bp. The protein-coding genes of the O. volvulus mitochondrial genome exhibit a striking codon bias, with 15/20 amino acids having a single codon preference greater than 70%. Intraspecific variation in both the nuclear and mitochondrial genomes appears to be quite limited, consistent with the hypothesis that O. volvulus has suffered a genetic bottleneck in the recent past.}, } @article {pmid10731564, year = {2000}, author = {Williams, SA and Lizotte-Waniewski, MR and Foster, J and Guiliano, D and Daub, J and Scott, AL and Slatko, B and Blaxter, ML}, title = {The filarial genome project: analysis of the nuclear, mitochondrial and endosymbiont genomes of Brugia malayi.}, journal = {International journal for parasitology}, volume = {30}, number = {4}, pages = {411-419}, doi = {10.1016/s0020-7519(00)00014-x}, pmid = {10731564}, issn = {0020-7519}, mesh = {Animals ; Brugia malayi/*genetics ; Chromosome Mapping ; DNA, Mitochondrial/chemistry ; Expressed Sequence Tags ; Filariasis/genetics ; *Genome ; Humans ; Wolbachia/genetics ; }, abstract = {The Filarial Genome Project (FGP) was initiated in 1994 under the auspices of the World Health Organisation. Brugia malayi was chosen as the model organism due to the availability of all life cycle stages for the construction of cDNA libraries. To date, over 20000 cDNA clones have been partially sequenced and submitted to the EST database (dbEST). These ESTs define approximately 7000 new Brugia genes. Analysis of the EST dataset provides useful information on the expression pattern of the most abundantly expressed Brugia genes. Some highly expressed genes have been identified that are expressed in all stages of the parasite's life cycle, while other highly expressed genes appear to be stage-specific. To elucidate the structure of the Brugia genome and to provide a basis for comparison to the Caenorhabditis elegans genome, the FGP is also constructing a physical map of the Brugia chromosomes and is sequencing genomic BAC clones. In addition to the nuclear genome, B. malayi possesses two other genomes: the mitochondrial genome and the genome of a bacterial endosymbiont. Eighty percent of the mitochondrial genome of B. malayi has been sequenced and is being compared to mitochondrial sequences of other nematodes. The bacterial endosymbiont genome found in B. malayi is closely related to the Wolbachia group of rickettsia-like bacteria that infects many insect species. A set of overlapping BAC clones is being assembled to cover the entire bacterial genome. Currently, half of the bacterial genome has been assembled into four contigs. A consortium has been established to sequence the entire genome of the Brugia endosymbiont. The sequence and mapping data provided by the FGP is being utilised by the nematode research community to develop a better understanding of the biology of filarial parasites and to identify new vaccine candidates and drug targets to aid the elimination of human filariasis.}, } @article {pmid10692011, year = {2000}, author = {Bordenstein, SR and Werren, JH}, title = {Do Wolbachia influence fecundity in Nasonia vitripennis?.}, journal = {Heredity}, volume = {84 (Pt 1)}, number = {}, pages = {54-62}, doi = {10.1046/j.1365-2540.2000.00637.x}, pmid = {10692011}, issn = {0018-067X}, mesh = {Animals ; Female ; Fertility ; Host-Parasite Interactions ; Male ; Wasps/genetics/*microbiology/physiology ; Wolbachia/pathogenicity/*physiology ; }, abstract = {This paper reports the influence of a vertically transmitted symbiont, Wolbachia, on host fitness in the parasitic wasp, Nasonia vitripennis. We measured fecundities of uninfected strains and strains infected with either two Wolbachia variants (wAv,wBv) or one (wAv or wBv). Preliminary tests suggested that double-infected females produce more offspring on average than uninfected females. However, further studies failed to yield consistent fitness effects. To control for host genetic effects, the genotype of the double-infected and uninfected strain was 'replaced' with three different host genetic backgrounds by introgression. Contrary to previous results, we found no convincing evidence for positive fitness effects of Wolbachia in Nasonia vitripennis, once host genetic background was controlled for. It can be concluded that under the experimental design used here, the fecundity effects associated with Wolbachia in N. vitripennis are small or absent.}, } @article {pmid10670955, year = {2000}, author = {Jiggins, FM and Hurst, GD and Majerus, ME}, title = {Sex-ratio-distorting Wolbachia causes sex-role reversal in its butterfly host.}, journal = {Proceedings. Biological sciences}, volume = {267}, number = {1438}, pages = {69-73}, pmid = {10670955}, issn = {0962-8452}, mesh = {Animals ; Butterflies/genetics/microbiology/*physiology ; Female ; Male ; Reproduction ; Sex Ratio ; Sexual Behavior, Animal ; Wolbachia/*physiology ; }, abstract = {Sex-role-reversed mating systems in which females compete for males and males may be choosy are usually associated with males investing more than females in offspring. We report that sex-role reversal may also be caused by selfish genetic elements which distort the sex ratio towards females. Some populations of the butterflies Acraea encedon and Acraea encedana are extremely female biased because over 90% of females are infected with a Wolbachia bacterium that is maternally inherited and kills male embryos. Many females in these populations are virgins suggesting that their reproductive success may be limited by access to males. These females form lekking swarms at landmarks in which females exhibit behaviours which we interpret as functioning to solicit matings from males. The hypothesis that female A. encedon swarm in order to mate is supported by the finding that, in release recapture experiments, mated females tend to leave the swarm while unmated females remained. This behaviour is a sex-role-reversed form of a common mating system in insects in which males form lekking swarms at landmarks and compete for females. Female lekking swarms are absent from less female-biased populations and here the butterflies are instead associated with resources in the form of the larval food plant.}, } @article {pmid10668862, year = {1999}, author = {van Opijnen, T and Breeuwer, JA}, title = {High temperatures eliminate Wolbachia, a cytoplasmic incompatibility inducing endosymbiont, from the two-spotted spider mite.}, journal = {Experimental & applied acarology}, volume = {23}, number = {11}, pages = {871-881}, pmid = {10668862}, issn = {0168-8162}, mesh = {Administration, Oral ; Animals ; Anti-Bacterial Agents/administration & dosage/pharmacology ; Crosses, Genetic ; DNA, Bacterial/genetics/isolation & purification ; Female ; Hot Temperature ; Male ; Mites/*microbiology/physiology ; Polymerase Chain Reaction ; Sex Ratio ; *Symbiosis ; Wolbachia/drug effects/genetics/*physiology ; }, abstract = {Wolbachia can induce cytoplasmic incompatibility (CI) in the arrhenotokous two-spotted spider mite between uninfected females and infected males. Cytoplasmic incompatibility is expressed through a male-biased sex ratio and a low hatchability, and can be suppressed by removing Wolbachia from spider mites reared on a diet with antibiotics. Here we investigated whether heat-treatment can elimate Wolbachia from infected mites. Using a PCR assay with a Wolbachia-specific primer pair (ftsZ), and by standard crosses, we were able to show that 71 per cent of the mites had lost the Wolbachia infection after rearing the infected population at 32+/-0.5 degrees C for four generations. The infection could be completely removed when mites were reared at 32+/-0.5 degrees C for six generations. Curing through high temperatures could be one of the reasons why mixed infected/uninfected populations occur in the field. An additional consequence of rearing mites at 32+/-0.5 degrees C was the shortened development time. The effect of environmental temperature on the abundance of Wolbachia and possible behavioural consequences for the spider mite are discussed.}, } @article {pmid10655228, year = {2000}, author = {Presgraves, DC}, title = {A genetic test of the mechanism of Wolbachia-induced cytoplasmic incompatibility in Drosophila.}, journal = {Genetics}, volume = {154}, number = {2}, pages = {771-776}, pmid = {10655228}, issn = {0016-6731}, support = {GM-51932/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Crosses, Genetic ; Drosophila melanogaster/embryology/*microbiology ; Female ; Male ; Spermatozoa/*microbiology ; Wolbachia/*pathogenicity ; }, abstract = {Cytoplasmic bacteria of the genus Wolbachia are best known as the cause of cytoplasmic incompatibility (CI): many uninfected eggs fertilized by Wolbachia-modified sperm from infected males die as embryos. In contrast, eggs of infected females rescue modified sperm and develop normally. Although Wolbachia cause CI in at least five insect orders, the mechanism of CI remains poorly understood. Here I test whether the target of Wolbachia-induced sperm modification is the male pronucleus (e.g., DNA or pronuclear proteins) or some extranuclear factor from the sperm required for embryonic development (e.g., the paternal centrosome). I distinguish between these hypotheses by crossing gynogenetic Drosophila melanogaster females to infected males. Gynogenetic females produce diploid eggs whose normal development requires no male pronucleus but still depends on extranuclear paternal factors. I show that when gynogenetic females are crossed to infected males, uniparental progeny with maternally derived chromosomes result. This finding shows that Wolbachia impair the male pronucleus but no extranuclear component of the sperm.}, } @article {pmid10652551, year = {2000}, author = {Rokas, I}, title = {Wolbachia as a speciation agent.}, journal = {Trends in ecology & evolution}, volume = {15}, number = {2}, pages = {44-45}, doi = {10.1016/s0169-5347(99)01783-8}, pmid = {10652551}, issn = {1872-8383}, } @article {pmid10652077, year = {2000}, author = {Malloch, G and Fenton, B and Butcher, RD}, title = {Molecular evidence for multiple infections of a new subgroup of Wolbachia in the European raspberry beetle Byturus tomentosus.}, journal = {Molecular ecology}, volume = {9}, number = {1}, pages = {77-90}, doi = {10.1046/j.1365-294x.2000.00836.x}, pmid = {10652077}, issn = {0962-1083}, mesh = {Animals ; Bacterial Proteins/genetics ; Base Sequence ; Coleoptera/classification/*microbiology ; *Cytoskeletal Proteins ; Ecosystem ; Europe ; Genes, Bacterial/genetics ; Genetic Variation/genetics ; Molecular Sequence Data ; Phylogeny ; Point Mutation/genetics ; Polymerase Chain Reaction ; Recombination, Genetic ; Sequence Alignment ; Symbiosis ; Wolbachia/*classification/genetics/*physiology ; }, abstract = {Wolbachia, a group of maternally inherited intracellular parasitic bacteria, alter host reproduction, including the induction of thelytokous parthenogenesis, feminization of genetic males, son killing and, most commonly, the induction of cytoplasmic incompatibility (CI), in a diverse array of arthropods. CI can result in infertility and has attracted attention because of its potential in biological control and as an agent in speciation. Although there has been some analysis of overall infection rates in arthropods and within individual insect orders, there has been little exploration of within-species variation. In this study, primers specific for the ftsZ gene of Wolbachia were used to amplify it from different geographical samples of the European raspberry beetle (Byturus tomentosus), confirming the presence of Wolbachia. More than 99% of UK individuals were found to be infected with Wolbachia and 97% of these B. tomentosus beetles harboured multiple infections. Preliminary analysis of B. tomentosus beetles from continental European populations revealed a lower level of infection (24%) than those from the UK. Phylogenetic analysis using the ftsZ DNA sequences places Wolbachia from B. tomentosus into a new clade (Abt) within the A division, with some revisions to the existing Wolbachia phylogeny.}, } @article {pmid10620236, year = {1999}, author = {Fukatsu, T}, title = {Acetone preservation: a practical technique for molecular analysis.}, journal = {Molecular ecology}, volume = {8}, number = {11}, pages = {1935-1945}, doi = {10.1046/j.1365-294x.1999.00795.x}, pmid = {10620236}, issn = {0962-1083}, mesh = {*Acetone ; Animals ; Aphids/*genetics/*microbiology ; Buchnera/*physiology ; DNA/chemistry/isolation & purification ; Electrophoresis, Polyacrylamide Gel ; Ethanol ; Immunohistochemistry ; In Situ Hybridization ; Proteins/chemistry/isolation & purification ; RNA/chemistry/isolation & purification ; Specimen Handling ; Symbiosis ; *Tissue Preservation ; }, abstract = {In attempts to establish a convenient and reliable method for field collection and archival preservation of insects and their endosymbiotic microorganisms for molecular analysis, acetone, ethanol, and other organic solvents were tested for DNA preservability of the pea aphid Acyrthosiphon pisum and its intracellular symbiotic bacterium Buchnera sp. After 6 months' storage, not only the band of high-molecular-size DNA but also the bands of rRNA were well preserved in acetone, ethanol, 2-propanol, diethyl ether and ethyl acetate. Polymerase chain reaction (PCR) assays confirmed that the DNA of both the insects and their symbionts was well preserved in these solvents. In contrast, methanol and chloroform showed poor DNA preservability. When water-containing series of acetone and ethanol were examined for DNA preservability, acetone was apparently more robust against water contamination than ethanol. Considering that most biological materials contain high amounts of water, acetone may be a more recommendable preservative for DNA analysis than ethanol which has been widely used for this purpose. The DNA of various insects could be preserved in acetone at room temperature in good condition for several years. In addition to the DNA of the host insects, the DNA of their endosymbionts, including Buchnera and other mycetocyte symbionts, Wolbachia, and gut bacteria, was amplified by PCR after several years of acetone storage. The RNA and protein of the pea aphid and its endosymbiont were also preserved for several years in acetone. After 2 years' storage in acetone, proteins of A. pisum could be analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, and the endosymbiotic bacteria were successfully detected by immunohistochemistry and in situ hybridization on the tissue sections.}, } @article {pmid18811258, year = {1997}, author = {Clancy, DJ and Hoffmann, AA}, title = {Behavior of Wolbachia endosymbionts from Drosophila simulans in Drosophila serrata, a novel host.}, journal = {The American naturalist}, volume = {149}, number = {5}, pages = {975-988}, doi = {10.1086/286033}, pmid = {18811258}, issn = {0003-0147}, abstract = {Many species harbor the incompatibility-inducing microbe Wolbachia, a maternally inherited endoparasite that causes reduced egg hatch in crosses between infected males and uninfected females. Infected females are immune to this effect, which gives them a relative fitness advantage that results in the spread of the infection. The strength of incompatibility, fitness deficits associated with the infection, and transmission rate from mother to offspring largely determine the rate and extent of spread of Wolbachia in a population. We transferred Wolbachia from Drosophila simulans to Drosophila serrata, a novel host, and compared parameter estimates with those from three naturally occurring Drosophila-Wolbachia associations believed to be of different ages. Transfected D. serrata showed strong incompatibility, low transmission efficiency, and an associated fitness deficit, and they would probably be unable to spread in nature. The comparisons generally supported the predicted evolution of a host-Wolbachia association. The parameters peculiar to any given host-Wolbachia association may determine whether the microbial strain can spread in that host.}, } @article {pmid15012323, year = {1997}, author = {Werren, JH}, title = {Biology of Wolbachia.}, journal = {Annual review of entomology}, volume = {42}, number = {}, pages = {587-609}, doi = {10.1146/annurev.ento.42.1.587}, pmid = {15012323}, issn = {0066-4170}, abstract = {Wolbachia are a common and widespread group of bacteria found in reproductive tissues of arthropods. These bacteria are transmitted through the cytoplasm of eggs and have evolved various mechanisms for manipulating reproduction of their hosts, including induction of reproductive incompatibility, parthenogenesis, and feminization. Wolbachia are also transmitted horizontally between arthropod species. Significant recent advances have been made in the study of these interesting microorganisms. In this paper, Wolbachia biology is reviewed, including their phylogeny and distribution, mechanisms of action, population biology and evolution, and biological control implications. Potential directions for future research are also discussed.}, } @article {pmid15275347, year = {1995}, author = {O'Neill, SL}, title = {Wolbachia pipientis: symbiont or parasite?.}, journal = {Parasitology today (Personal ed.)}, volume = {11}, number = {5}, pages = {168-169}, doi = {10.1016/0169-4758(95)80146-4}, pmid = {15275347}, issn = {0169-4758}, } @article {pmid14144721, year = {1964}, author = {SUITOR, EC}, title = {STUDIES ON THE CELL ENVELOPE OF WOLBACHIA PERSICA.}, journal = {The Journal of infectious diseases}, volume = {114}, number = {}, pages = {125-134}, doi = {10.1093/infdis/114.2.125}, pmid = {14144721}, issn = {0022-1899}, mesh = {*Bacitracin ; *Bacteria ; *Cell Membrane ; Chick Embryo ; *Culture Media ; *Cycloserine ; *DNA ; *Deoxyribonucleases ; *Detergents ; *Electrons ; *Microscopy ; *Microscopy, Electron ; *Muramidase ; *Osmosis ; *Penicillins ; *Pharmacology ; *Polymyxins ; *Research ; *Ribonucleases ; *Trypsin ; *Tyrothricin ; *Ultrasonics ; *Wolbachia ; }, } @article {pmid14118048, year = {1964}, author = {WEISS, E and NEPTUNE, EM and DAVIES, JA}, title = {LIPID METABOLISM OF THE RICKETTSIALIKE MICROORGANISM WOLBACHIA PERSICA. III. COMPARISON WITH OTHER METABOLIC ACTIVITIES.}, journal = {The Journal of infectious diseases}, volume = {114}, number = {}, pages = {50-54}, doi = {10.1093/infdis/114.1.50}, pmid = {14118048}, issn = {0022-1899}, mesh = {*Bacteria ; *Carbon Isotopes ; *Glucose ; *Lipid Metabolism ; *Palmitic Acid ; *Research ; *Serine ; *Wolbachia ; }, } @article {pmid14118047, year = {1964}, author = {NEPTUNE, EM and WEISS, E and DAVIES, JA}, title = {LIPID METABOLISM OF THE RICKETTSIALIKE MICROORGANISM WOLBACHIA PERSICA. II. STUDIES WITH LABELED NONLIPID SUBSTRATES.}, journal = {The Journal of infectious diseases}, volume = {114}, number = {}, pages = {45-49}, doi = {10.1093/infdis/114.1.45}, pmid = {14118047}, issn = {0022-1899}, mesh = {*Acetates ; *Bacteria ; *Carbon Isotopes ; *Escherichia coli ; *Fatty Acids ; *Glucose ; *Glutamine ; *Glycerol ; *Glycerophosphates ; *Lecithins ; *Lipid Metabolism ; *Metabolism ; *Palmitic Acid ; *Phosphatidylcholines ; *Phosphatidylethanolamines ; *Phospholipids ; *Research ; *Serine ; *Wolbachia ; }, } @article {pmid14118046, year = {1964}, author = {NEPTUNE, EM and WEISS, E and DAVIES, JA and SUITOR, EC}, title = {LIPID METABOLISM OF THE RICKETTSIALIKE MICROORGANISM WOLBACHIA PERSICA. I. INCORPORATION OF LONG CHAIN FATTY ACIDS INTO PHOSPHATIDES.}, journal = {The Journal of infectious diseases}, volume = {114}, number = {}, pages = {39-44}, doi = {10.1093/infdis/114.1.39}, pmid = {14118046}, issn = {0022-1899}, mesh = {*Bacteria ; *Carbon Isotopes ; Chick Embryo ; *Chromatography ; *Escherichia coli ; *Fatty Acids ; *Glucose ; *Lecithins ; *Lipid Metabolism ; *Manometry ; *Palmitic Acid ; *Phosphatidylcholines ; *Phosphatidylethanolamines ; *Phospholipids ; *Research ; *Wolbachia ; }, } @article {pmid14005862, year = {1962}, author = {WEISS, E and MYERS, WF and SUITOR, EC and NEPTUNE, EM}, title = {Respiration of a rickettsialike microorganism, Wolbachia persica.}, journal = {The Journal of infectious diseases}, volume = {110}, number = {}, pages = {155-164}, doi = {10.1093/infdis/110.2.155}, pmid = {14005862}, issn = {0022-1899}, mesh = {*Respiration ; Wolbachia/*metabolism ; }, } @article {pmid10612622, year = {1999}, author = {Cui, L and Chang, SH and Strickman, D and Rowton, E}, title = {Frequency of Wolbachia infection in laboratory and field sand fly (Diptera: Psychodidae) populations.}, journal = {Journal of the American Mosquito Control Association}, volume = {15}, number = {4}, pages = {571-572}, pmid = {10612622}, issn = {8756-971X}, mesh = {Animals ; Membrane Proteins/genetics ; Polymerase Chain Reaction ; Population Dynamics ; Psychodidae/*parasitology ; Wolbachia/genetics/isolation & purification/*pathogenicity ; }, abstract = {Using a polymerase chain reaction (PCR) assay with primers designed for the Wolbachia 16S rRNA and outer surface protein (wsp) gene, we screened 11 laboratory colonies and 4 field samples of 6 sand fly species for Wolbachia infection. Infection was only detected in 3 laboratory colonies of Phlebotomus papatasi Scopoli, originally collected in Israel, Egypt, and Saudi Arabia.}, } @article {pmid10605113, year = {1999}, author = {Vavre, F and Fleury, F and Lepetit, D and Fouillet, P and Boulétreau, M}, title = {Phylogenetic evidence for horizontal transmission of Wolbachia in host-parasitoid associations.}, journal = {Molecular biology and evolution}, volume = {16}, number = {12}, pages = {1711-1723}, doi = {10.1093/oxfordjournals.molbev.a026084}, pmid = {10605113}, issn = {0737-4038}, mesh = {Animals ; Bacterial Outer Membrane Proteins/*genetics ; Base Sequence ; Data Interpretation, Statistical ; Drosophila/*microbiology ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {Endosymbiotic Wolbachia infect a number of arthropod species in which they can affect the reproductive system. While maternally transmitted, unlike mitochondria their molecular phylogeny does not parallel that of their hosts. This strongly suggests horizontal transmission among species, the mechanisms of which remain unknown. Such transfers require intimate between-species relationships, and thus host-parasite associations are outstandingly appropriate for study. Here, we demonstrate that hymenopteran parasitoids of frugivorous Drosophila species are especially susceptible to Wolbachia infection. Of the five common European species, four proved to be infected; furthermore, multiple infections are common, with one species being doubly infected and two triply infected (first report). Phylogenetic statuses of the Wolbachia infecting the different species of the community have been studied using the gene wsp, a highly variable gene recently described. This study reveals exciting similarities between the Wolbachia variants found in parasitoids and their hosts. These arguments strongly support the hypothesis of frequent natural Wolbachia transfers into other species and open a new field for genetic exchanges among species, especially in host-parasitoid associations.}, } @article {pmid10591206, year = {1999}, author = {Hadfield, SJ and Axton, JM}, title = {Germ cells colonized by endosymbiotic bacteria.}, journal = {Nature}, volume = {402}, number = {6761}, pages = {482}, doi = {10.1038/45002}, pmid = {10591206}, issn = {0028-0836}, mesh = {Animals ; Cell Polarity ; Drosophila/*microbiology ; Female ; Genes, Insect ; Ovum/*microbiology ; Symbiosis ; Wolbachia/*physiology ; }, } @article {pmid10587943, year = {1999}, author = {Slatko, BE and O'Neill, SL and Scott, AL and Werren, JL and Blaxter, ML}, title = {The Wolbachia Genome Consortium.}, journal = {Microbial & comparative genomics}, volume = {4}, number = {3}, pages = {161-165}, doi = {10.1089/omi.1.1999.4.161}, pmid = {10587943}, issn = {1090-6592}, mesh = {Biotechnology ; *Genome, Bacterial ; Research ; Sequence Analysis, DNA ; Wolbachia/*genetics/physiology ; }, } @article {pmid10587942, year = {1999}, author = {O'Neill, SL}, title = {Wolbachia: why these bacteria are important to genome research.}, journal = {Microbial & comparative genomics}, volume = {4}, number = {3}, pages = {159}, doi = {10.1089/omi.1.1999.4.159}, pmid = {10587942}, issn = {1090-6592}, mesh = {Animals ; Arthropods/microbiology ; *Genome, Bacterial ; *Research ; Wolbachia/*genetics/*physiology ; Wuchereria/microbiology ; }, } @article {pmid10583549, year = {1999}, author = {Rigaud, T and Moreau, J and Juchault, P}, title = {Wolbachia infection in the terrestrial isopod oniscus asellus: sex ratio distortion and effect on fecundity.}, journal = {Heredity}, volume = {83}, number = {# (Pt 4)}, pages = {469-475}, doi = {10.1038/sj.hdy.6885990}, pmid = {10583549}, issn = {1365-2540}, abstract = {Maternally inherited Wolbachia bacteria are widespread in arthropods where they are responsible for various reproductive alterations. In terrestrial isopods (woodlice), Wolbachia may induce feminization or cytoplasmic incompatibility (CI), but their effect remains unknown in most host species. To increase our understanding of host/symbiont interactions in terrestrial isopods, the effect of Wolbachia was investigated in the oniscidean Oniscus asellus, mainly to discriminate between feminization and CI. The Wolbachia infection was not linked with a CI phenomenon, but females infected with Wolbachia produced female-biased broods compared with uninfected females. The fecundity of infected females was slightly lower than that of uninfected, but the number of young at the adult stage was similar between the two female categories. The experimental transfer of the symbiont into uninfected strains showed that Wolbachia was responsible for the feminization of a number of genetic males. In female-biased broods, Wolbachia were vertically transmitted to around 88% of the offspring, but the transmission rate was lower in the few male-biased progenies. The feminizing activity of these symbionts was not systematic, as many phenotypic males were infected. These results contrasted with what is known in another woodlouse species, and indicated that feminization has evolved in different ways in terrestrial isopods.}, } @article {pmid10556723, year = {1999}, author = {Vandekerckhove, TT and Watteyne, S and Willems, A and Swings, JG and Mertens, J and Gillis, M}, title = {Phylogenetic analysis of the 16S rDNA of the cytoplasmic bacterium Wolbachia from the novel host Folsomia candida (Hexapoda, Collembola) and its implications for wolbachial taxonomy.}, journal = {FEMS microbiology letters}, volume = {180}, number = {2}, pages = {279-286}, doi = {10.1111/j.1574-6968.1999.tb08807.x}, pmid = {10556723}, issn = {0378-1097}, mesh = {Animals ; DNA, Bacterial/genetics ; DNA, Ribosomal/*genetics ; Female ; Insecta/*microbiology ; Microscopy, Electron ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods ; RNA, Ribosomal, 16S/*genetics ; Wolbachia/*classification/*genetics/ultrastructure ; }, abstract = {Wolbachia pipientis are intracellular, transovarially inherited alpha-Proteobacteria in invertebrates. Four major Wolbachia groups exist: A, B (contained in divergent arthropods), C and D (harbored by Nematoda). By means of transmission electron microscopy, we observed Wolbachia-like bacteria in a primitive insect, Folsomia candida (Hexapoda, Collembola, Isotomidae). 16S rDNA analysis proved them to constitute a novel lineage, henceforth named group E, in the wolbachial phylogenetic tree. It shares 97.8% 16S rDNA homology with its nearest neighbors, groups A and B, which diverged from it more recently. We propose (i) a new taxon E for the Wolbachia strain in F. candida, (ii) that the single-described Wolbachia pipientis fall apart into at least three species: C, D and the large E-A-B complex. F. candida's group E Wolbachia rekindle the question about invasive capacities of free-living ancestral wolbachiae and horizontal transfer.}, } @article {pmid10555290, year = {1999}, author = {Clark, MA and Moran, NA and Baumann, P}, title = {Sequence evolution in bacterial endosymbionts having extreme base compositions.}, journal = {Molecular biology and evolution}, volume = {16}, number = {11}, pages = {1586-1598}, doi = {10.1093/oxfordjournals.molbev.a026071}, pmid = {10555290}, issn = {0737-4038}, mesh = {AT Rich Sequence ; Amino Acid Substitution ; Base Sequence ; Buchnera/*genetics ; DNA Primers ; *Evolution, Molecular ; }, abstract = {A major limitation on ability to reconstruct bacterial evolution is the lack of dated ancestors that might be used to evaluate and calibrate molecular clocks. Vertically transmitted symbionts that have cospeciated with animal hosts offer a firm basis for calibrating sequence evolution in bacteria, since fossils of the hosts can be used to date divergence events. Sequences for a functionally diverse set of genes have been obtained for bacterial endosymbionts (Buchnera) from two pairs of aphid host species, each pair diverging 50-70 MYA. Using these dates and estimated numbers of Buchnera generations per year, we calculated rates of base substitution for neutral and selected sites of protein-coding genes and overall rates for rRNA genes. Buchnera shows homogeneity among loci with regard to synonymous rate. The Buchnera synonymous rate is about twice that for low-codon-bias genes of Escherichia coli-Salmonella typhimurium on an absolute timescale, and fourfold higher on a generational timescale. Nonsynonymous substitutions show a greater rate disparity in favor of Buchnera, a result consistent with a genomewide decrease in selection efficiency in Buchnera. Ratios of synonymous to nonsynonymous substitutions differ for the two pairs of Buchnera, indicating that selection efficiency varies among lineages. Like numerous other intracellular bacteria, such as Rickettsia and Wolbachia, Buchnera has accumulated amino acids with codons rich in A or T. Phylogenetic reconstruction of amino acid replacements indicates that replacements yielding increased A + T predominated early in the evolution of Buchnera, with the trend slowing or stopping during the last 50 Myr. This suggests that base composition in Buchnera has approached a limit enforced by selective constraint acting on protein function.}, } @article {pmid10547686, year = {1999}, author = {Stouthamer, R and Breeuwer, JA and Hurst, GD}, title = {Wolbachia pipientis: microbial manipulator of arthropod reproduction.}, journal = {Annual review of microbiology}, volume = {53}, number = {}, pages = {71-102}, doi = {10.1146/annurev.micro.53.1.71}, pmid = {10547686}, issn = {0066-4227}, mesh = {Animals ; Arthropods/*microbiology/*physiology ; Biological Evolution ; Female ; Male ; Phenotype ; Reproduction/genetics/physiology ; Wolbachia/*physiology ; }, abstract = {The alpha-proteobacterium Wolbachia pipientis is a very common cytoplasmic symbiont of insects, crustaceans, mites, and filarial nematodes. To enhance its transmission, W. pipientis has evolved a large scale of host manipulations: parthenogenesis induction, feminization, and male killing. W. pipientis's most common effect is a crossing incompatibility between infected males and uninfected females. Little is known about the genetics and biochemistry of these symbionts because of their fastidious requirements. The affinity of W. pipientis for the microtubules associated with the early divisions in eggs may explain some of their effects. Such inherited microorganisms are thought to have been major factors in the evolution of sex determination, eusociality, and speciation. W. pipientis isolates are also of interest as vectors for the modification of wild insect populations, in the improvement of parasitoid wasps in biological pest control, and as a new method for interfering with diseases caused by filarial nematodes.}, } @article {pmid10511685, year = {1999}, author = {Taylor, MJ and Hoerauf, A}, title = {Wolbachia bacteria of filarial nematodes.}, journal = {Parasitology today (Personal ed.)}, volume = {15}, number = {11}, pages = {437-442}, doi = {10.1016/s0169-4758(99)01533-1}, pmid = {10511685}, issn = {0169-4758}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Arthropods/microbiology ; Brugia malayi/*microbiology/ultrastructure ; Female ; Filariasis/drug therapy/immunology/microbiology ; Immunohistochemistry ; Male ; Microscopy, Electron ; Onchocerca volvulus/*microbiology/ultrastructure ; Phylogeny ; Symbiosis/genetics ; Wolbachia/*growth & development ; Wuchereria bancrofti/*microbiology/ultrastructure ; }, abstract = {The finding that the intracellular bacteria of filarial nematodes are related to the Wolbachia symbionts of arthropods has generated great interest. Here, Mark Taylor and Achim Hoerauf review recent studies by several groups on the structure, distribution and phylogeny of these endosymbionts, and discuss the potential role for these bacteria in filarial disease and as a target for chemotherapy.}, } @article {pmid10511680, year = {1999}, author = {Bandi, C and Slatko, B and O'Neill, SL}, title = {Wolbachia genomes and the many faces of symbiosis.}, journal = {Parasitology today (Personal ed.)}, volume = {15}, number = {11}, pages = {428-429}, doi = {10.1016/s0169-4758(99)01543-4}, pmid = {10511680}, issn = {0169-4758}, mesh = {Animals ; Arthropods/*microbiology/physiology ; Female ; *Genome, Bacterial ; Male ; Nematoda/*microbiology/physiology ; Sequence Analysis, DNA/methods ; *Symbiosis ; Wolbachia/chemistry/*genetics/physiology ; }, } @article {pmid10469257, year = {1999}, author = {Van Meer, MM and Witteveldt, J and Stouthamer, R}, title = {Phylogeny of the arthropod endosymbiont Wolbachia based on the wsp gene.}, journal = {Insect molecular biology}, volume = {8}, number = {3}, pages = {399-408}, doi = {10.1046/j.1365-2583.1999.83129.x}, pmid = {10469257}, issn = {0962-1075}, mesh = {Animals ; Arthropods/parasitology ; Bacterial Outer Membrane Proteins/*genetics ; Base Sequence ; DNA Primers ; DNA, Bacterial ; *Genes, Bacterial ; Molecular Sequence Data ; Phenotype ; Phylogeny ; Rickettsiaceae/*classification/genetics ; Symbiosis ; }, abstract = {Bacteria of the genus Wolbachia (Rickettsiae) are widespread in arthropods and can induce cytoplasmic incompatibility (CI), thelytoky (T) or feminization (F) in their host. Recent research on the wsp gene of mainly CI inducing Wolbachia has shown that this gene evolves at a much faster rate than previously sequenced genes such as 16S or ftsZ. As a result this gene appears to be very useful in subdividing the Wolbachia and twelve groups have been distinguished to date. Here we extend the Wolbachia wsp data set with fifteen T-Wolbachia, one F-Wolbachia and three other CI-Wolbachia strains. The results showed: (i) the addition of seven groups; (ii) no relation between host phenotype and Wolbachia phylogenetic position; and (iii) possible horizontal Wolbachia transfer between the moth Ephestia kuehniella and its parasitoid Trichogramma spp.}, } @article {pmid10447705, year = {1999}, author = {Marcad, I and Souty-Grosset, C and Bouchon, D and Rigaud, T and Raimond, R}, title = {Mitochondrial DNA variability and wolbachia infection in two sibling woodlice species.}, journal = {Heredity}, volume = {83 (Pt 1)}, number = {}, pages = {71-78}, doi = {10.1038/sj.hdy.6885380}, pmid = {10447705}, issn = {1365-2540}, abstract = {Several morphological races and subspecies have been described and later included within the terrestrial isopod species Porcellionides pruinosus. During our study of this species, we have worked on specimens from France, Greece, Tunisia and Reunion island. Laboratory crosses have revealed two separate groups of populations: French populations (four localities) in one group, and those from Tunisia, Reunion island and Greece in the other. French individuals were reproductively isolated from those of the other populations. We have undertaken a survey of mitochondrial DNA (mtDNA) polymorphism in these seven populations. We observed two groups of mitotypes corresponding to the two groups of populations. Interfertility experiments between populations and the mitochondrial genetic distances between mitotypes both suggest the presence of two different species, one in France and one in Greece, Tunisia and Reunion island. The two species harbour, respectively, two different Wolbachia lines. Another feature of the molecular genetic analysis was the apparent mitochondrial monomorphism in the French populations and the low variability in the other three populations. The result can be related to the possibility of Wolbachia-induced genetic hitchhiking in these populations.}, } @article {pmid10430591, year = {1999}, author = {Rigaud, T and Bouchon, D and Souty-Grosset, C and Raimond, R}, title = {Mitochondrial DNA polymorphism, sex ratio distorters and population genetics in the isopod Armadillidium vulgare.}, journal = {Genetics}, volume = {152}, number = {4}, pages = {1669-1677}, pmid = {10430591}, issn = {0016-6731}, mesh = {Animals ; Crustacea/*genetics/microbiology/physiology ; DNA, Mitochondrial/*genetics ; Female ; France ; Genetics, Population ; Male ; Plasmids/*genetics ; Rickettsiaceae/*genetics ; *Sex Ratio ; }, abstract = {Two maternally inherited sex ratio distorters (SRD) impose female-biased sex ratios on the wood louse Armadillidium vulgare by feminizing putative males. These SRD are (i) an intracytoplasmic bacterium of the genus Wolbachia, and (ii) another non-Mendelian element of unknown nature: the f element. Mitochondrial DNA variation was investigated in A. vulgare field populations to trace the evolution of host-SRD relationships and to investigate the effect of SRD on host cytoplasmic polymorphism. The Wolbachia endosymbionts showed no polymorphism in their ITS2 sequence and were associated with two closely related mitochondrial types. This situation probably reflects a single infection event followed by a slight differentiation of mitochondria. There was no association between the f element and a given mitochondrial type, which may confirm the fact that this element can be partially paternally transmitted. The spreading of a maternally inherited SRD in a population should reduce the mitochondrial diversity by a hitchhiking process. In A. vulgare, however, a within-population mtDNA polymorphism was often found, because of the deficient spread of Wolbachia and the partial paternal inheritance of the f element. The analysis of molecular variance indicated that A. vulgare populations are genetically structured, but without isolation by distance.}, } @article {pmid10419978, year = {1999}, author = {Sun, LV and Babaratsas, A and Savakis, C and O'Neill, SL and Bourtzis, K}, title = {Gene organization of the dnaA region of Wolbachia.}, journal = {Journal of bacteriology}, volume = {181}, number = {15}, pages = {4708-4710}, pmid = {10419978}, issn = {0021-9193}, support = {AI 40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics ; Chromosome Mapping ; DNA-Binding Proteins/genetics ; Drosophila/microbiology ; Glutamate-Ammonia Ligase/genetics ; Insecta/microbiology ; Open Reading Frames ; Rickettsiaceae/classification/*genetics ; }, abstract = {The dnaA region of Wolbachia, an intracellular bacterial parasite of insects, is unique. A glnA cognate was found upstream of the dnaA gene, while neither of the two open reading frames detected downstream of dnaA has any homologue in the database. This unusual gene arrangement may reflect requirements associated with the unique ecological niche this agent occupies.}, } @article {pmid10413661, year = {1999}, author = {Masui, S and Kamoda, S and Sasaki, T and Ishikawa, H}, title = {The first detection of the insertion sequence ISW1 in the intracellular reproductive parasite Wolbachia.}, journal = {Plasmid}, volume = {42}, number = {1}, pages = {13-19}, doi = {10.1006/plas.1999.1407}, pmid = {10413661}, issn = {0147-619X}, mesh = {Amino Acid Sequence ; Animals ; Base Sequence ; DNA Primers/genetics ; DNA Transposable Elements/*genetics ; DNA, Bacterial/genetics ; Female ; Gryllidae/microbiology/physiology ; Molecular Sequence Data ; Phylogeny ; Reproduction ; Rickettsiaceae/*genetics/pathogenicity ; Salmonella typhimurium/genetics ; Sequence Homology, Amino Acid ; }, abstract = {Wolbachia are maternally inherited intracellular rickettsia-like bacteria known to infect a wide range of arthropods. They are associated with a number of different reproductive phenotypes in their hosts, such as cytoplasmic incompatibility, parthenogenesis, and feminization. We report on a novel insertion sequence (IS), ISW1, which was identified in the region downstream of groEL of a Wolbachia strain, wTai. The 573-bp-long ISW1 sequence is the first IS element observed in this organism, displays significant similarity to IS200, and lacks terminal inverted repeats. There were more than 20 copies of ISW1 on the chromosome of wTai. Sequence analysis of nine distinct ISW1 copies and their flanking regions showed that the copies were identical and suggested that ISW1 has no preference for its insertion sites. Possible roles of ISW1 in the adaptation of Wolbachia to intracellular environments and in various reproductive alterations caused by this bacterium are discussed.}, } @article {pmid10411634, year = {1999}, author = {Martin, G and Sorokine, O and Moniatte, M and Bulet, P and Hetru, C and Van Dorsselaer, A}, title = {The structure of a glycosylated protein hormone responsible for sex determination in the isopod, Armadillidium vulgare.}, journal = {European journal of biochemistry}, volume = {262}, number = {3}, pages = {727-736}, doi = {10.1046/j.1432-1327.1999.00442.x}, pmid = {10411634}, issn = {0014-2956}, mesh = {Amino Acid Sequence ; Animals ; Carbohydrate Conformation ; Chromatography, High Pressure Liquid ; Crustacea/*chemistry/genetics/physiology ; Dimerization ; Glycoproteins/*chemistry/isolation & purification/physiology ; *Gonadal Hormones ; Gonadal Steroid Hormones/*chemistry/isolation & purification/physiology ; Male ; Molecular Sequence Data ; Molecular Weight ; Peptide Fragments/chemistry ; Polysaccharides/chemistry ; *Sex Determination Processes ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Sulfides ; }, abstract = {Two glycoforms (AH1 and AH2) of androgenic hormone, and its corresponding hormone precursor derived from HPLC-purified androgenic gland extract from the woodlouse Armadillidium vulgare were fully characterized by microsequencing and mass spectrometry. The amino-acid sequences of the two glycoforms were identical; they consist of two peptide chains, A and B, of 29 and 44 amino acids, respectively, with chain A carrying one N-glycosylated moiety on Asn18. The two chains are linked by two disulfide bridges. Glycoforms were only differentiated by the size and heterogeneity of the glycan chain. The androgenic hormone precursor (16.5 kDa) was shown to contain the sequence of chains A and B from the androgenic hormone, connected by a C-peptide (50 amino acids). These results were confirmed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analysis performed on a single hypertrophied androgenic gland. When injected into young females, both glycoforms of the androgenic hormone were able to override genetic sex-determination. In invertebrates, there is no other example where sex-differentiation is controlled by a protein hormone that is not synthesized by the gonads but by a special gland. A functional comparison with two other hormones which are believed to play a role in sex determination, i.e. ecdysone in insects and anti-Müllerian hormone in mammals, is presented. Work is in progress to clone and characterize the gene encoding androgenic hormone, moreover special attention is devoted to its regulatory regions, putative targets for the Wolbachia action.}, } @article {pmid10390640, year = {1999}, author = {McGraw, EA and O'Neill, SL}, title = {Evolution of Wolbachia pipientis transmission dynamics in insects.}, journal = {Trends in microbiology}, volume = {7}, number = {7}, pages = {297-302}, doi = {10.1016/s0966-842x(99)01531-0}, pmid = {10390640}, issn = {0966-842X}, support = {AI40620/AI/NIAID NIH HHS/United States ; T32 AI07404/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Cytoplasm/physiology ; Drosophila/*microbiology/physiology ; Female ; Male ; Reproduction ; Rickettsiaceae/*genetics/*physiology ; }, abstract = {Wolbachia pipientis is an intracellular bacterial parasite of arthropods that enhances its transmission by manipulating host reproduction, most commonly by inducing cytoplasmic incompatibility. The discovery of isolates with modified cytoplasmic incompatibility phenotypes and others with novel virulence properties is an indication of the potential breadth of evolutionary strategies employed by Wolbachia.}, } @article {pmid10383683, year = {1999}, author = {Rousset, F and Braig, HR and O'Neill, SL}, title = {A stable triple Wolbachia infection in Drosophila with nearly additive incompatibility effects.}, journal = {Heredity}, volume = {82 (Pt 6)}, number = {}, pages = {620-627}, doi = {10.1046/j.1365-2540.1999.00501.x}, pmid = {10383683}, issn = {0018-067X}, support = {AI 40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*microbiology/physiology ; Female ; Male ; Reproduction/physiology ; Rickettsiaceae/*physiology ; }, abstract = {Drosophila simulans strains infected with three different Wolbachia strains were generated by experimental injection of a third symbiont into a naturally double-infected strain. This transfer led to a substantial increase in total Wolbachia density in the host strain. Each of the three symbionts was stably transmitted in the presence of the other two. Triple-infected males were incompatible with double-infected females. No evidence was obtained for interference between modification effects of the different Wolbachia strains in males. Some incompatibility was observed between triple-infected males and females. However, this incompatibility reaction is not a specific property of triple-infected flies, because it was also observed in double-infected strains.}, } @article {pmid10380108, year = {1999}, author = {Turelli, M and Hoffmann, AA}, title = {Microbe-induced cytoplasmic incompatibility as a mechanism for introducing transgenes into arthropod populations.}, journal = {Insect molecular biology}, volume = {8}, number = {2}, pages = {243-255}, doi = {10.1046/j.1365-2583.1999.820243.x}, pmid = {10380108}, issn = {0962-1075}, mesh = {Animals ; *Arthropods ; Cytoplasm ; *Models, Genetic ; Rickettsia/*genetics ; *Transgenes ; }, abstract = {Many arthropods are infected with maternally transmitted, intracellular bacteria of the genus Wolbachia. These infections often produce 'cytoplasmic incompatibility' (CI)--reduced egg-hatch frequencies when uninfected females mate with infected males or when males and females carrying different Wolbachia strains mate. Because infected females often enjoy a fitness advantage--they are effectively immune to any effects from males carrying the same Wolbachia strain--Wolbachia and associated cytoplasmic elements can spread rapidly through natural populations. Wolbachia might therefore drive transgenes associated with disease control or pest abatement into populations. In this paper, simple mathematical analyses are presented of three alternative strategies for 'CI drive'. The analyses reveal which parameters must be estimated in order to predict population dynamics, and they demonstrate stringent requirements for initially driving and/or maintaining transgenes in target populations.}, } @article {pmid10380102, year = {1999}, author = {Plantard, O and Rasplus, JY and Mondor, G and Le Clainche, I and Solignac, M}, title = {Distribution and phylogeny of Wolbachia inducing thelytoky in Rhoditini and 'Aylacini' (Hymenoptera: Cynipidae).}, journal = {Insect molecular biology}, volume = {8}, number = {2}, pages = {185-191}, doi = {10.1046/j.1365-2583.1999.820185.x}, pmid = {10380102}, issn = {0962-1075}, mesh = {Animals ; Base Sequence ; DNA, Complementary ; Molecular Sequence Data ; Phylogeny ; Rickettsia/*classification/genetics ; Wasps/*microbiology ; }, abstract = {Wolbachia are endosymbiotic bacteria responsible for thelytoky in several parasitoid hymenopteran genera. After finding these micro-organisms in some populations of Diplolepis spinosissimae (Hymenoptera: Cynipidae) where they are responsible for thelytoky through gamete duplication, we searched for Wolbachia spp. using specific PCR primers in nineteen other species of the Rhoditini tribe (rose gallwasps) and eight species of the 'Aylacini' tribe (gallwasps associated with herbaceous plants). Wolbachia were found in twelve Rhoditini species and four 'Aylacini' species. The most infected species have very few males (spanandry) and the thelytoky of infected species/arrhenotoky of uninfected species is confirmed by previous research based on the sex of the offspring of virgin females. Phylogenetic analyses based on the partial Wolbachia ftsZ gene sequences indicate that some strains associated with closely related gallwasps are phylogenetically distant, suggesting that cynipids have been affected by several infection events. In contrast, the five infected European species of Diplolepis harbour the same strain of Wolbachia.}, } @article {pmid10376278, year = {1998}, author = {Genchi, C and Sacchi, L and Bandi, C and Venco, L}, title = {Preliminary results on the effect of tetracycline on the embryogenesis and symbiotic bacteria (Wolbachia) of Dirofilaria immitis. An update and discussion.}, journal = {Parassitologia}, volume = {40}, number = {3}, pages = {247-249}, pmid = {10376278}, issn = {0048-2951}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Dirofilaria immitis/drug effects/*embryology ; Dirofilariasis/parasitology ; Dog Diseases/parasitology ; Dogs ; Female ; Morphogenesis/drug effects ; Rickettsia/*drug effects ; Symbiosis/*drug effects ; Tetracycline/*pharmacology ; }, abstract = {The distribution and phylogeny of Wolbachia in filarial species suggests that these endosymbiotic bacteria may be important in the biology of their filarial hosts. An experiment to falsify this hypothesis would be to treat filarial worms with antibiotics which are active against intracellular bacteria. Indeed, it has already been shown that tetracycline treatment inhibits development in a model filarial species (Brugia pahangi) at different stages of the life cycle, in both mosquito and mammalian hosts. Here we discuss these previous data and present new results on the effect of tetracycline on the embryogenesis of the canine filaria Dirofilaria immitis.}, } @article {pmid10359795, year = {1999}, author = {Heddi, A and Grenier, AM and Khatchadourian, C and Charles, H and Nardon, P}, title = {Four intracellular genomes direct weevil biology: nuclear, mitochondrial, principal endosymbiont, and Wolbachia.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {96}, number = {12}, pages = {6814-6819}, pmid = {10359795}, issn = {0027-8424}, mesh = {Biological Evolution ; DNA, Mitochondrial ; Gene Expression Regulation ; Genome, Bacterial ; *Genome, Plant ; Molecular Sequence Data ; Oryza/*genetics ; Symbiosis/*genetics ; }, abstract = {Cell physiology in the weevil Sitophilus oryzae is coordinated by three integrated genomes: nuclear, mitochondrial, and the "S. oryzae principal endosymbiont" (SOPE). SOPE, a cytoplasmic bacterium (2 x 10(3) bacteria per specialized bacteriocyte cell and 3 x 10(6) bacteria per weevil) that belongs to the proteobacteria gamma3-subgroup, is present in all weevils studied. We discovered a fourth prokaryotic genome in somatic and germ tissues of 57% of weevil strains of three species, S. oryzae, Sitophilus zeamais, and Sitophilus granarius, distributed worldwide. We assigned this Gram-negative prokaryote to the Wolbachia group (alpha-proteobacteria), on the basis of 16S rDNA sequence and fluorescence in situ DNA-RNA hybridization (FISH). Both bacteria, SOPE and Wolbachia, were selectively eliminated by combined heat and antibiotic treatments. Study of bacteria involvement in this insect's genetics and physiology revealed that SOPE, which induces the specific differentiation of the bacteriocytes, increases mitochondrial oxidative phosphorylation through the supply of pantothenic acid and riboflavin. Elimination of this gamma3-proteobacterium impairs many physiological traits. By contrast, neither the presence nor the absence of Wolbachia significantly affects the weevil's physiology. Wolbachia, disseminated throughout the body cells, is in particularly high density in the germ cells, where it causes nucleocytoplasmic incompatibility. The coexistence of two distinct types of intracellular proteobacteria at different levels of symbiont integration in insects illustrates the genetic complexity of animal tissue. Furthermore, evolutionary timing can be inferred: first nucleocytoplasm, then mitochondria, then SOPE, and finally Wolbachia. Symbiogenesis, the genetic integration of long-term associated members of different species, in the weevil appears to be a mechanism of speciation (with Wolbachia) and provides a means for animals to acquire new genes that permit better adaptation to the environment (with SOPE).}, } @article {pmid10221636, year = {1999}, author = {Bandi, C and McCall, JW and Genchi, C and Corona, S and Venco, L and Sacchi, L}, title = {Effects of tetracycline on the filarial worms Brugia pahangi and Dirofilaria immitis and their bacterial endosymbionts Wolbachia.}, journal = {International journal for parasitology}, volume = {29}, number = {2}, pages = {357-364}, doi = {10.1016/s0020-7519(98)00200-8}, pmid = {10221636}, issn = {0020-7519}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Brugia/*drug effects/microbiology ; Dirofilaria/*drug effects/microbiology ; Dogs ; Elephantiasis, Filarial/veterinary ; Female ; Gerbillinae/parasitology ; Microscopy, Electron ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods ; Rickettsiaceae/*drug effects/genetics/isolation & purification ; Symbiosis ; Tetracycline/*pharmacology ; }, abstract = {Wolbachia endosymbiotic bacteria have been shown to be widespread among filarial worms and could thus play some role in the biology of these nematodes. Indeed, tetracycline has been shown to inhibit both the development of adult worms from third-stage larvae and the development of the microfilaraemia in jirds infected with Brugia pahangi. The possibility that these effects are related to the bacteriostatic activity of tetracycline on Wolbachia symbionts should be considered. Here we show that tetracycline treatment is very effective in blocking embryo development in two filarial nematodes, B. pahangi and Dirofilaria immitis. Embryo degeneration was documented by TEM, while the inhibition of the transovarial transmission of Wolbachia was documented by PCR. Phylogenetic analysis on the ssrDNA sequence of the Wolbachia of B. pahangi confirms that the phylogeny of the bacterial endosymbionts is consistent with that of the host worms. The possibility that tetracycline inhibition of embryo development in B. pahangi and D. immitis is determined by cytoplasmic incompatibility is discussed.}, } @article {pmid10209097, year = {1999}, author = {Heath, BD and Butcher, RD and Whitfield, WG and Hubbard, SF}, title = {Horizontal transfer of Wolbachia between phylogenetically distant insect species by a naturally occurring mechanism.}, journal = {Current biology : CB}, volume = {9}, number = {6}, pages = {313-316}, doi = {10.1016/s0960-9822(99)80139-0}, pmid = {10209097}, issn = {0960-9822}, mesh = {Animals ; DNA, Bacterial/isolation & purification ; Female ; Host-Parasite Interactions ; Insecta/*microbiology/parasitology ; Male ; Microscopy, Fluorescence ; Ovary/microbiology ; Polymerase Chain Reaction ; Pupa/microbiology ; Rickettsiaceae/*physiology ; Species Specificity ; Wasps/*microbiology/physiology ; }, abstract = {Wolbachia is a genus of alpha-proteobacteria found in obligate intracellular association with a wide variety of arthropods, including an estimated 10-20% of all insect species [1]. Wolbachia represents one of a number of recently identified 'reproductive parasites' [2] which manipulate the reproduction of their hosts in ways that enhance their own transmission [3] [4] [5] [6] [7] [8] [9]. The influence of Wolbachia infection on the dynamics of host populations has focused considerable interest on its possible role in speciation through reproductive isolation [3] [10] [11] and as an agent of biological control [2] [12] [13]. Although Wolbachia normally undergoes vertical transmission through the maternal line of its host population [14], there is compelling evidence from molecular phylogenies that extensive horizontal (intertaxon) transmission must have occurred [1] [9] [15] [16] [17]. Some of the best candidate vectors for the horizontal transmission of Wolbachia are insect parasitoids [15], which comprise around 25% of all insect species and attack arthropods from an enormous range of taxa [18]. In this study, we used both fluorescence microscopy and PCR amplification with Wolbachia-specific primers to show that Wolbachia can be transmitted to a parasitic wasp (Leptopilina boulardi) from its infected host (Drosophila simulans) and subsequently undergo diminishing vertical transmission in this novel host species. These results are, to our knowledge, the first to reveal a natural horizontal transfer route for Wolbachia between phylogenetically distant insect species.}, } @article {pmid10196738, year = {1999}, author = {Dobson, SL and Bourtzis, K and Braig, HR and Jones, BF and Zhou, W and Rousset, F and O'Neill, SL}, title = {Wolbachia infections are distributed throughout insect somatic and germ line tissues.}, journal = {Insect biochemistry and molecular biology}, volume = {29}, number = {2}, pages = {153-160}, doi = {10.1016/s0965-1748(98)00119-2}, pmid = {10196738}, issn = {0965-1748}, support = {AI07404-07/AI/NIAID NIH HHS/United States ; AI34355/AI/NIAID NIH HHS/United States ; AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Blotting, Western ; DNA, Bacterial/*analysis ; Drosophila/*microbiology ; Gonads/microbiology ; Infectious Disease Transmission, Vertical ; Polymerase Chain Reaction ; Rickettsia/*genetics/pathogenicity ; Rickettsia Infections/transmission ; Tissue Distribution ; }, abstract = {Wolbachia are intracellular microorganisms that form maternally-inherited infections within numerous arthropod species. These bacteria have drawn much attention, due in part to the reproductive alterations that they induce in their hosts including cytoplasmic incompatibility (CI), feminization and parthenogenesis. Although Wolbachia's presence within insect reproductive tissues has been well described, relatively few studies have examined the extent to which Wolbachia infects other tissues. We have examined Wolbachia tissue tropism in a number of representative insect hosts by western blot, dot blot hybridization and diagnostic PCR. Results from these studies indicate that Wolbachia are much more widely distributed in host tissues than previously appreciated. Furthermore, the distribution of Wolbachia in somatic tissues varied between different Wolbachia/host associations. Some associations showed Wolbachia disseminated throughout most tissues while others appeared to be much more restricted, being predominantly limited to the reproductive tissues. We discuss the relevance of these infection patterns to the evolution of Wolbachia/host symbioses and to potential applied uses of Wolbachia.}, } @article {pmid10098265, year = {1999}, author = {Van Meer, MM and Stouthamer, R}, title = {Cross-order transfer of Wolbachia from Muscidifurax uniraptor (Hymenoptera: Pteromalidae) to Drosophila simulans (Diptera: Drosophilidae).}, journal = {Heredity}, volume = {82 (Pt 2)}, number = {}, pages = {163-169}, doi = {10.1038/sj.hdy.6884610}, pmid = {10098265}, issn = {0018-067X}, mesh = {Alphaproteobacteria/*growth & development ; Animals ; Bacterial Proteins/genetics ; Crosses, Genetic ; Cytoplasm/genetics ; *Cytoskeletal Proteins ; Drosophila/*genetics/microbiology ; Female ; Fertility ; Hymenoptera/*genetics/microbiology ; Male ; Microinjections ; Ovum/growth & development ; Polymerase Chain Reaction ; }, abstract = {Bacteria of the genus Wolbachia are widespread in arthropods and can induce different effects on the host such as cytoplasmic incompatibility (CI), thelytoky (T) or feminization (F). In some Wolbachia-infected hosts, no effect (N) has been found. Successful transfer of Wolbachia by microinjection from one host to an uninfected one has been established with CI, F, N-Wolbachia but not with T-Wolbachia. In this paper a transfer experiment of T-Wolbachia from the parasitoid Muscidifurax uniraptor to Drosophila simulans is described. The infection could be detected in the new host for several generations by polymerase chain reaction (PCR). However, no specific effects on the host were detected, and the bacteria were not stably maintained.}, } @article {pmid10092480, year = {1999}, author = {Taylor, MJ and Bilo, K and Cross, HF and Archer, JP and Underwood, AP}, title = {16S rDNA phylogeny and ultrastructural characterization of Wolbachia intracellular bacteria of the filarial nematodes Brugia malayi, B. pahangi, and Wuchereria bancrofti.}, journal = {Experimental parasitology}, volume = {91}, number = {4}, pages = {356-361}, doi = {10.1006/expr.1998.4383}, pmid = {10092480}, issn = {0014-4894}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aedes ; Animals ; Base Sequence ; Brugia malayi/*microbiology/ultrastructure ; Brugia pahangi/*microbiology/ultrastructure ; DNA, Bacterial/chemistry ; DNA, Ribosomal/*chemistry ; Elephantiasis, Filarial/parasitology ; Female ; Gerbillinae ; Humans ; Male ; Mice ; Mice, SCID ; Molecular Sequence Data ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/*classification/genetics/ultrastructure ; Wuchereria bancrofti/*microbiology/ultrastructure ; }, } @article {pmid10030026, year = {1999}, author = {van Meer, MM and van Kan, FJ and Stouthamer, R}, title = {Spacer 2 region and 5S rDNA variation of Wolbachia strains involved in cytoplasmic incompatibility or sex-ratio distortion in arthropods.}, journal = {Letters in applied microbiology}, volume = {28}, number = {1}, pages = {17-22}, doi = {10.1046/j.1365-2672.1999.00476.x}, pmid = {10030026}, issn = {0266-8254}, mesh = {Animals ; Arthropods/*microbiology/physiology ; Bacterial Proteins/genetics ; Base Sequence ; *Cytoskeletal Proteins ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/*genetics ; Female ; Genetic Variation ; Male ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/methods ; RNA, Ribosomal, 5S/*genetics ; Reproduction/physiology ; Rickettsiaceae/*genetics/isolation & purification ; Sex Ratio ; }, abstract = {Bacteria in the genus Wolbachia are widespread in arthropods and can induce sex-ratio distortion or cytoplasmic incompatibility in their hosts. The phylogeny of Wolbachia has been studied using 16S rDNA and the cell cycle gene ftsZ, but sequence variation of those genes is limited. The spacer 2 region (SR2) was amplified to determine whether this region would improve phylogenetic resolution. The SR2 of Wolbachia is 66 bp long, shows higher variation than ftsZ and has very low homology with closely related bacteria. Due to the small length of SR2 of Wolbachia, little phylogenetic information could be retrieved.}, } @article {pmid9927181, year = {1999}, author = {Cheng, Q and Aksoy, S}, title = {Tissue tropism, transmission and expression of foreign genes in vivo in midgut symbionts of tsetse flies.}, journal = {Insect molecular biology}, volume = {8}, number = {1}, pages = {125-132}, doi = {10.1046/j.1365-2583.1999.810125.x}, pmid = {9927181}, issn = {0962-1075}, support = {NIH-AI-34033/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteria/genetics ; Digestive System/microbiology ; Female ; Gene Expression ; *Genes, Bacterial ; Hemolymph/microbiology ; Recombination, Genetic ; Symbiosis ; Tissue Distribution ; Tsetse Flies/*microbiology ; }, abstract = {Tsetse flies (Diptera: Glossinidae) harbour three different symbiotic organisms in addition to the pathogenic African trypanosomes they transmit. The two gut-associated symbionts (primary, P; secondary, S) are enteric and are nutritionally required, whereas the third microorganism Wolbachia (family Rickettsiaceae) affects the reproductive biology of the insects it infects. The bacteriome-associated P-symbiont (Wigglesworthia glossinidia) displays a concordant phylogeny with its host tsetse species, whereas midgut S-symbionts characterized from distant tsetse have identical 16S rDNA sequences and therefore may either represent recent independent acquisitions or horizontal transfer between species. The S-symbionts have been cultured in vitro and a genetic transformation system has been developed. Here we report on their density and tissue tropism in different species (G. m. morsitans, G. p. palpalis, G. austeni and G. brevipalpis) and on their maternal route of transmission to tsetse progeny. Using a bacterium-specific PCR-assay, the S-symbionts were found primarily in the midgut, haemolymph, milk gland and in G. palpalis also in salivary glands of teneral flies. In older flies these infections were found to spread to other tissues including muscle, testes and fat body. The S-symbionts were transformed to express the marker gene product, Green Fluorescent Protein (GFP) in vitro. When the recombinant symbionts were introduced into the haemoceal of fertile female flies via intrathoracic microinjection, they were detected in the intrauterine progeny, indicating that haemolymph may provide a possible route for their transmission. The implications of these results for symbiont-host interactions and for transgenic strategies in tsetse are discussed.}, } @article {pmid9927175, year = {1999}, author = {Vavre, F and Girin, C and Boulétreau, M}, title = {Phylogenetic status of a fecundity-enhancing Wolbachia that does not induce thelytoky in Trichogramma.}, journal = {Insect molecular biology}, volume = {8}, number = {1}, pages = {67-72}, doi = {10.1046/j.1365-2583.1999.810067.x}, pmid = {9927175}, issn = {0962-1075}, mesh = {Animals ; Bacterial Proteins/genetics ; Base Sequence ; *Cytoskeletal Proteins ; DNA, Complementary ; Female ; Fertility ; Male ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S ; Rickettsia/classification/*genetics ; Wasps/*microbiology/physiology ; }, abstract = {Wolbachia are widespread bacteria which infect a number of species of insects and other arthropods. They manipulate the reproduction of their hosts at their own advantage. In Trichogramma species all Wolbachia known so far induce thelytoky and form a monophyletic group in the B subdivision of Wolbachia. Here we show that some strains of the arrhenotokous species Trichogramma bourarachae harbour Wolbachia symbionts that locate in the A subdivision, and which do not induce thelytoky. Although the symbiont of T. bourarachae is closely related to Wolbachia that induce cytoplasmic incompatibility in other insects, no cytoplasmic incompatibilities were found in crosses involving infected and uninfected strains. In T. bourarachae the presence of this Wolbachia is associated with a higher fecundity of strains. Our results strongly suggest that Wolbachia are involved in this increased fecundity. Theoretical models on the evolution of host-Wolbachia interaction predict that a reduced effect on reproduction can be selected for if cost of infection is reduced. The effect in T. bourarachae should illustrate this prediction.}, } @article {pmid9921679, year = {1998}, author = {Bandi, C and Anderson, TJ and Genchi, C and Blaxter, ML}, title = {Phylogeny of Wolbachia in filarial nematodes.}, journal = {Proceedings. Biological sciences}, volume = {265}, number = {1413}, pages = {2407-2413}, pmid = {9921679}, issn = {0962-8452}, mesh = {Animals ; Bacterial Proteins/classification/genetics ; Cattle ; *Cytoskeletal Proteins ; DNA, Bacterial ; DNA, Ribosomal/analysis ; Dogs ; Filarioidea/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S ; Rickettsiaceae/*classification/genetics ; }, abstract = {Intracellular bacteria have been observed in various species of filarial nematodes (family Onchocercidae). The intracellular bacterium of the canine filaria Dirofilaria immitis has been shown to be closely related to Wolbachia, a rickettsia-like micro-organism that is widespread among arthropods. However, the relationships between endosymbionts of different filariae, and between these and the arthropod wolbachiae, appear not to have been studied. To address these issues we have examined ten species of filarial nematodes for the presence of Wolbachia. For nine species, all samples examined were PCR positive using primers specific for the ftsZ gene of Wolbachia. For one species, the examined samples were PCR negative. Sequences of the amplified ftsZ gene fragments of filarial wolbachiae fall into two clusters (C and D), which are distinct from the A and B clusters recognized for arthropod wolbachiae. These four lineages (A-D) are related in a star-like phylogeny, with higher nucleotide divergence observed between C and D wolbachiae than that observed between A and B wolbachiae. In addition, within each of the two lineages of filarial wolbachiae, the phylogeny of the symbionts is consistent with the host phylogeny. Thus, there is no evidence for recent Wolbachia transmission between arthropods and nematodes. Endosymbiont 16S ribosomal DNA sequences from a subset of filarial species support these findings.}, } @article {pmid9884329, year = {1999}, author = {Hoerauf, A and Nissen-Pähle, K and Schmetz, C and Henkle-Dührsen, K and Blaxter, ML and Büttner, DW and Gallin, MY and Al-Qaoud, KM and Lucius, R and Fleischer, B}, title = {Tetracycline therapy targets intracellular bacteria in the filarial nematode Litomosoides sigmodontis and results in filarial infertility.}, journal = {The Journal of clinical investigation}, volume = {103}, number = {1}, pages = {11-18}, pmid = {9884329}, issn = {0021-9738}, mesh = {Animals ; Bacterial Proteins/analysis ; Dipetalonema/drug effects ; Filariasis/drug therapy ; Filarioidea/drug effects/*microbiology ; Immunohistochemistry ; Infertility ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Phylogeny ; RNA, Ribosomal, 16S/analysis ; Rats ; Rickettsia/*drug effects ; Tetracycline/*pharmacology ; }, abstract = {Intracellular bacteria have been described in several species of filarial nematodes, but their relationships with, and effects on, their nematode hosts have not previously been elucidated. In this study, intracellular bacteria were observed in tissues of the rodent parasite Litomosoides sigmodontis by transmission electron microscopy and by immunohistochemistry using antiendobacterial heat shock protein-60 antisera. Molecular phylogenetic analysis of the bacterial 16S ribosomal RNA gene, isolated by PCR, showed a close relationship to the rickettsial Wolbachia endobacteria of arthropods and to other filarial intracellular bacteria. The impact of tetracycline therapy of infected rodents on L. sigmodontis development was analyzed in order to understand the role(s) these bacteria might play in filarial biology. Tetracycline therapy, when initiated with L. sigmodontis infection, eliminated the bacteria and resulted in filarial growth retardation and infertility. If initiated after microfilarial development, treatment reduced filarial fertility. Treatment with antibiotics not affecting rickettsial bacteria did not inhibit filarial development. Acanthocheilonema viteae filariae were shown to lack intracellular bacteria and to be insensitive to tetracycline. These results suggest a mutualistic interaction between the intracellular bacteria and the filarial nematode. Investigation of such a mutualism in endobacteria-containing human filariae is warranted for a potential chemotherapeutic exploitation.}, } @article {pmid9873076, year = {1999}, author = {Chen, X and Li, S and Aksoy, S}, title = {Concordant evolution of a symbiont with its host insect species: molecular phylogeny of genus Glossina and its bacteriome-associated endosymbiont, Wigglesworthia glossinidia.}, journal = {Journal of molecular evolution}, volume = {48}, number = {1}, pages = {49-58}, doi = {10.1007/pl00006444}, pmid = {9873076}, issn = {0022-2844}, support = {NIH-AI-34033/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Base Sequence ; Cloning, Molecular ; DNA, Ribosomal/genetics ; Enterobacteriaceae/classification/*genetics ; *Evolution, Molecular ; Molecular Sequence Data ; *Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sequence Homology, Nucleic Acid ; Symbiosis/*genetics ; Tsetse Flies/classification/*genetics ; }, abstract = {Many arthropods with restricted diets rely on symbiotic associations for full nutrition and fecundity. Tsetse flies (Diptera: Glossinidae) harbor three symbiotic organisms in addition to the parasitic African trypanosomes they transmit. Two of these microorganisms reside in different gut cells, while the third organism is harbored in reproductive tissues and belongs to the genus Wolbachia. The primary symbiont (genus Wigglesworthia glossinidia) lives in differentiated epithelial cells (bacteriocytes) which form an organ (bacteriome) in the anterior gut, while the secondary (S) symbionts are present in midgut cells. Here we have characterized the phylogeny of Wigglesworthia based on their 16S rDNA sequence analysis from eight species representing the three subgenera of Glossina: Austenina (=fusca group), Nemorhina (=palpalis group), and Glossina (=morsitans group). Independently, the ribosomal DNA internal transcribed spacer-2 (ITS-2) regions from these species were analyzed. The analysis of Wigglesworthia indicated that they form a distinct lineage in the gamma subdivision of Proteobacteria and display concordance with their host insect species. The trees generated by parsimony confirmed the monophyletic taxonomic placement of Glossina, where fusca group species formed the deepest branch followed by morsitans and palpalis groups, respectively. The placement of the species Glossina austeni by both the traditional morphological and biochemical criteria has been controversial. Results presented here, based on both the ITS-2 and the symbiont 16S rDNA sequence analysis, suggest that Glossina austeni should be placed into a separate fourth subgenus, Machadomyia, which forms a sister-group relationship with the morsitans group species.}, } @article {pmid9851608, year = {1998}, author = {Henkle-Dührsen, K and Eckelt, VH and Wildenburg, G and Blaxter, M and Walter, RD}, title = {Gene structure, activity and localization of a catalase from intracellular bacteria in Onchocerca volvulus.}, journal = {Molecular and biochemical parasitology}, volume = {96}, number = {1-2}, pages = {69-81}, doi = {10.1016/s0166-6851(98)00109-1}, pmid = {9851608}, issn = {0166-6851}, mesh = {Amino Acid Sequence ; Animals ; Catalase/analysis/chemistry/*genetics/metabolism ; Female ; *Genes, Bacterial ; Genes, rRNA ; Immunoenzyme Techniques ; Male ; Microfilariae/enzymology/microbiology ; Microscopy, Electron ; Molecular Sequence Data ; Onchocerca volvulus/enzymology/growth & development/*microbiology/ultrastructure ; Open Reading Frames ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Recombinant Proteins/chemistry/metabolism ; Rickettsiaceae/enzymology/*genetics/ultrastructure ; Sequence Alignment ; }, abstract = {Within the context of studies on the antioxidant enzymes in Onchocerca volvulus, DNA clones encoding catalase (CAT) were isolated from an O. volvulus adult lambda zapII cDNA library. Analysis of their nucleotide and encoded amino acid sequences revealed that they derive from intracellular bacteria, rather than the O. volvulus nuclear genome. The endobacterial CAT gene was found to lie in a gene cluster, followed by a ferritin gene and an excinuclease gene. The endobacterial CAT gene encodes a functional enzyme capable of detoxifying H2O2, demonstrated by producing an active recombinant protein in an E. coli expression system. The purified 54 kDa protein has CAT activity over a broad pH range, with a specific activity of 103,000 +/- 3000 U mg(-1). The optical spectrum of the endobacterial CAT shows that it is a ferric haem-containing protein with a Soret band at 405 nm. To investigate the phylogeny of the intracellular bacterium in O. volvulus, a segment of the 16S rRNA gene was amplified from total genomic DNA by a polymerase chain reaction using universal eubacterial primers. A phylogenetic analysis of the O. volvulus-derived 16S rRNA sequence revealed that the endobacterium belongs to a distinct Wolbachia clade of the order Rickettsiales. Onchocercomata and biopsies containing different onchocercal species were immunohistochemically stained using polyclonal antibodies raised against the recombinant endobacterial CAT. CAT was detected in the endobacteria in the hypodermis of adult male and female O. volvulus, O. ochengi, O. gibsoni and O. fasciata. The endobacterial enzyme was also detected in onchocercal oocytes and all embryonic stages including intrauterine microfilariae as well as skin microfilariae. O. volvulus thus harbours Wolbachia-like endosymbionts which are transovarially transmitted and show particular affinity for the hypodermal tissues of the lateral chords.}, } @article {pmid9819901, year = {1998}, author = {West, SA and Cook, JM and Werren, JH and Godfray, HC}, title = {Wolbachia in two insect host-parasitoid communities.}, journal = {Molecular ecology}, volume = {7}, number = {11}, pages = {1457-1465}, doi = {10.1046/j.1365-294x.1998.00467.x}, pmid = {9819901}, issn = {0962-1083}, mesh = {Animals ; Aphids/classification/*microbiology ; Bacterial Proteins/genetics ; *Cytoskeletal Proteins ; DNA, Bacterial/genetics ; Ecosystem ; Genes, Bacterial ; Lepidoptera/classification/*microbiology ; Phylogeny ; Rickettsiaceae/classification/genetics/*isolation & purification ; Species Specificity ; }, abstract = {Wolbachia form a group of intracellular bacteria that alter reproduction in their arthropod hosts. Two major phylogenetic subdivisions (A and B) of Wolbachia occur. Using a polymerase chain reaction assay we surveyed for the A and B group Wolbachia in 82 insect species from two temperate host-parasitoid communities (food webs) and a general collection of Lepidoptera caught at a light trap. One host-parasitoid community was based around leaf-mining Lepidoptera, and the other around Aphids. We found that: (i) 22.0% of insects sampled were infected with Wolbachia; and (ii) the prevalence and type (A or B) of Wolbachia infection differed significantly between communities and taxonomic groups. We obtained DNA sequences from the ftsZ gene for the group B Wolbachia found in six leaf-mining species and one of their parasitoids, as well as four of the Lepidoptera caught by a light trap. Taken together, the results of our survey and phylogenetic analyses of the sequence data suggest that host-parasitoid transfer of Wolbanchia is not the major route through which the species we have examined become infected. In addition, the Wolbachia strains observed in five leaf-mining species from the same genus were not closely related, indicating that transfer between species has not occurred due to a shared feeding niche or cospeciation.}, } @article {pmid9809424, year = {1998}, author = {Eremeeva, ME and Ching, WM and Wu, Y and Silverman, DJ and Dasch, GA}, title = {Western blotting analysis of heat shock proteins of Rickettsiales and other eubacteria.}, journal = {FEMS microbiology letters}, volume = {167}, number = {2}, pages = {229-237}, doi = {10.1111/j.1574-6968.1998.tb13233.x}, pmid = {9809424}, issn = {0378-1097}, support = {AI-17416/AI/NIAID NIH HHS/United States ; }, mesh = {Alphaproteobacteria/*chemistry/growth & development ; Animals ; Antibodies, Bacterial/immunology ; Antibodies, Monoclonal/immunology ; Bacteria/*chemistry/growth & development ; *Blotting, Western ; Cross Reactions ; Electrophoresis, Polyacrylamide Gel ; Heat-Shock Proteins/*chemistry/immunology ; Heat-Shock Response ; Mice ; Rabbits ; }, abstract = {Heat shock proteins (Hsp) of four Rickettsia species, three Bartonella species, two Ehrlichia species, Orientia tsutsugamushi and seventeen other eubacterial species were characterized by the enhanced chemiluminescence Western blotting (WB) technique with antibodies raised against recombinant Hsp from Escherichia coli and purified GroES from R. typhi. Although E. coli DnaK and GroEL have epitopes that are highly conserved among the homologous proteins found in Rickettsia, Ehrlichia, O. tsutsugamushi, Bartonella and other Proteobacteria, anti-E. coli DnaK and GroEL monoclonal antibodies (Dasch et al. (1990) Ann. N.Y. Acad. Sci. 590, 352-369) recognize less conserved epitopes. In contrast, epitopes on E. coli DnaJ, GrpE and GroES are much less conserved since anti-E. coli DnaJ, GrpE and GroES polyclonal antibodies did not recognize DnaJ, GrpE or GroES homologues in Rickettsia, Bartonella, Orientia, Ehrlichia and Legionella. Polyclonal antiserum prepared against GroES from R. typhi reacted strongly with purified 10 kDa GroES peptide from Rickettsia and Bartonella, and strongly bound to proteins of varying electrophoretic mobility from Wolbachia, Legionella, Proteus and Shigella flexneri and more weakly to other GroES homologues including that found in E. coli. Consequently, commercially available anti-DnaJ, anti-GrpE and anti-GroES polyclonal antibodies and anti-DnaK monoclonal antibody raised against their respective recombinant E. coli Hsp are not suitable for detection and identification of homologues of these proteins in a wide range of eubacteria.}, } @article {pmid9735249, year = {1998}, author = {Frank, SA}, title = {Dynamics of Cytoplasmic Incompatability with Multiple Wolbachia Infections.}, journal = {Journal of theoretical biology}, volume = {192}, number = {2}, pages = {213-218}, doi = {10.1006/jtbi.1998.0652}, pmid = {9735249}, issn = {1095-8541}, abstract = {Wolbachia infections occur in many arthropods. These matrilineally inherited bacteria cause cytoplasmic incompatibility, in which a cross produces no offspring when between an infected male and an uninfected female. Some populations harbour multiple Wolbachia strains. Females fail to produce offspring when crossed to a male with a strain that the female lacks. Prior theoretical work showed that a panmictic population cannot maintain polymorphism for different strains when each female carries only a single strain. A few authors suggested that doubly infected females can stabilize multistrain polymorphism, but conditions for invasion and location of stable equilibrium were not analysed in detail. For two strains, I describe the conditions under which a multiply infected class can spread. Spread of the doubly infected type stabilizes polymorphism of the singly infected classes. This analysis also suggests an interesting extension to higher multiplicity of infection. For an arbitrary number of strains, N, a panmictic population cannot maintain different classes with N-1 infections unless the class with N infections is also present. This pyramid of polymorphism may explain the puzzling diversity of incompatibility types observed in some Culex mosquitos. Multiple infection also has interesting consequences for the dynamics of spatial variation and reproductive isolation.Copyright 1998 Academic Press Limited}, } @article {pmid9725842, year = {1998}, author = {Poinsot, D and Bourtzis, K and Markakis, G and Savakis, C and Merçot, H}, title = {Wolbachia transfer from Drosophila melanogaster into D. simulans: Host effect and cytoplasmic incompatibility relationships.}, journal = {Genetics}, volume = {150}, number = {1}, pages = {227-237}, pmid = {9725842}, issn = {0016-6731}, support = {AI34355/AI/NIAID NIH HHS/United States ; AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/*physiology ; Drosophila/*microbiology ; Female ; Male ; Rickettsiaceae/isolation & purification ; Rickettsiaceae Infections/*physiopathology/transmission ; Species Specificity ; }, abstract = {Wolbachia are maternally transmitted endocellular bacteria causing a reproductive incompatibility called cytoplasmic incompatibility (CI) in several arthropod species, including Drosophila. CI results in embryonic mortality in incompatible crosses. The only bacterial strain known to infect Drosophila melanogaster (wDm) was transferred from a D. melanogaster isofemale line into uninfected D. simulans isofemale lines by embryo microinjections. Males from the resulting transinfected lines induce >98% embryonic mortality when crossed with uninfected D. simulans females. In contrast, males from the donor D. melanogaster line induce only 18-32% CI on average when crossed with uninfected D. melanogaster females. Transinfected D. simulans lines do not differ from the D. melanogaster donor line in the Wolbachia load found in the embryo or in the total bacterial load of young males. However, >80% of cysts are infected by Wolbachia in the testes of young transinfected males, whereas only 8% of cysts are infected in young males from the D. melanogaster donor isofemale line. This difference might be caused by physiological differences between hosts, but it might also involve tissue-specific control of Wolbachia density by D. melanogaster. The wDm-transinfected D. simulans lines are unidirectionally incompatible with strains infected by the non-CI expressor Wolbachia strains wKi, wMau, or wAu, and they are bidirectionally incompatible with strains infected by the CI-expressor Wolbachia strains wHa or wNo. However, wDm-infected males do not induce CI toward females infected by the CI-expressor strain wRi, which is found in D. simulans continental populations, while wRi-infected males induce partial CI toward wDm-infected females. This peculiar asymmetrical pattern could reflect an ongoing divergence between the CI mechanisms of wRi and wDm. It would also confirm other results indicating that the factor responsible for CI induction in males is distinct from the factor responsible for CI rescue in females.}, } @article {pmid9723877, year = {1998}, author = {Gottlieb, Y and Zchori-Fein, E and Faktor, O and Rosen, D}, title = {Phylogenetic analysis of parthenogenesis-inducing Wolbachia in the genus Aphytis (Hymenoptera: Aphelinidae).}, journal = {Insect molecular biology}, volume = {7}, number = {4}, pages = {393-396}, doi = {10.1046/j.1365-2583.1998.740393.x}, pmid = {9723877}, issn = {0962-1075}, mesh = {Animals ; Bacterial Proteins/genetics ; Base Sequence ; *Cytoskeletal Proteins ; DNA, Complementary ; Molecular Sequence Data ; *Parthenogenesis ; *Phylogeny ; Rickettsiaceae/*classification/genetics ; Wasps/*microbiology/physiology ; }, abstract = {Parthenogenesis-inducing intracellular bacteria of the genus Wolbachia are found in a variety of parasitoid wasp genera. The presence of Wolbachia in the uniparental Aphytis species A. lingnanensis Compere, A. diaspidis (Howard), A. chilensis Howard, and A. chrysomphali (Mercet) was tested using primers specific for the ftsZ gene. The symbiont was detected in all of these species. Wolbachia ftsZ genes that were sequenced from the four hosts show a high degree of similarity. Both the PCR with specific primers for group 'A' and phylogenetic analysis place these Wolbachia in group 'A'. The fact that the tested Aphytis species belong to different phylogenetic groups and harbour what seem to be almost identical Wolbachia, lends credence to the horizontal transmission hypothesis.}, } @article {pmid9721689, year = {1998}, author = {Wenseleers, T and Ito, F and Van Borm, S and Huybrechts, R and Volckaert, F and Billen, J}, title = {Widespread occurrence of the micro-organism Wolbachia in ants.}, journal = {Proceedings. Biological sciences}, volume = {265}, number = {1404}, pages = {1447-1452}, pmid = {9721689}, issn = {0962-8452}, mesh = {Animals ; Ants/*microbiology/*physiology ; Biological Evolution ; *Disorders of Sex Development ; Female ; Male ; Polymerase Chain Reaction ; Reproduction ; Rickettsiaceae/isolation & purification/pathogenicity/*physiology ; *Sex Determination Processes ; Social Behavior ; Species Specificity ; }, abstract = {For more than 20 years, sex allocation in hymenopteran societies has been a major topic in insect sociobiology. A recurring idea was that relatedness asymmetrics arising from their haplodiploid sex determination system would lead to various parent-offspring conflicts over optimal reproduction. A possible weakness of existing theory is that only interests of nuclear genes are properly accounted for. Yet, a diversity of maternally transmitted elements manipulate the reproduction of their host in many solitary arthropod groups. The bacterium Wolbachia is a striking example of such a selfish cytoplasmic element, with effects ranging from reproductive incompatibility between host strains, induction of parthenogenesis and feminization of males. This paper reports on a first PCR-based Wolbachia screening in ants. Out of 50 Indo-Australian species, 50% screened positive for an A-group strain. One of these species also harboured a B-group strain in a double infection. Various factors that might explain the unusually high incidence of Wolbachia in ants are discussed. In general, Wolbachia may represent a widespread and previously unrecognized party active in the conflicts of interest within social insect colonies.}, } @article {pmid9705412, year = {1998}, author = {Ohashi, N and Unver, A and Zhi, N and Rikihisa, Y}, title = {Cloning and characterization of multigenes encoding the immunodominant 30-kilodalton major outer membrane proteins of Ehrlichia canis and application of the recombinant protein for serodiagnosis.}, journal = {Journal of clinical microbiology}, volume = {36}, number = {9}, pages = {2671-2680}, pmid = {9705412}, issn = {0095-1137}, support = {R01 AI33123/AI/NIAID NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Bacterial Outer Membrane Proteins/biosynthesis/*genetics/immunology ; Base Sequence ; Cloning, Molecular/methods ; DNA Primers ; Dog Diseases/*diagnosis ; Dogs ; Ehrlichia/classification/*genetics/isolation & purification ; Ehrlichiosis/diagnosis/*veterinary ; Evolution, Molecular ; Genome, Bacterial ; Humans ; Introns ; Mice ; Molecular Sequence Data ; Molecular Weight ; *Multigene Family ; Open Reading Frames ; *Phylogeny ; Polymerase Chain Reaction/methods ; Sequence Alignment ; Sequence Homology, Amino Acid ; Serologic Tests/methods ; }, abstract = {A 30-kDa major outer membrane protein of Ehrlichia canis, the agent of canine ehrlichiosis, is the major antigen recognized by both naturally and experimentally infected dog sera. The protein cross-reacts with a serum against a recombinant 28-kDa protein (rP28), one of the outer membrane proteins of a gene (omp-1) family of Ehrlichia chaffeensis. Two DNA fragments of E. canis were amplified by PCR with two primer pairs based on the sequences of E. chaffeensis omp-1 genes, cloned, and sequenced. Each fragment contained a partial 30-kDa protein gene of E. canis. Genomic Southern blot analysis with the partial gene probes revealed the presence of multiple copies of these genes in the E. canis genome. Three copies of the entire gene (p30, p30-1, and p30a) were cloned and sequenced from the E. canis genomic DNA. The open reading frames of the two copies (p30 and p30-1) were tandemly arranged with an intergenic space. The three copies were similar but not identical and contained a semivariable region and three hypervariable regions in the protein molecules. The following genes homologous to three E. canis 30-kDa protein genes and the E. chaffeensis omp-1 family were identified in the closely related rickettsiae: wsp from Wolbachia sp. , p44 from the agent of human granulocytic ehrlichiosis, msp-2 and msp-4 from Anaplasma marginale, and map-1 from Cowdria ruminantium. Phylogenetic analysis among the three E. canis 30-kDa proteins and the major surface proteins of the rickettsiae revealed that these proteins are divided into four clusters and the two E. canis 30-kDa proteins are closely related but that the third 30-kDa protein is not. The p30 gene was expressed as a fusion protein, and the antibody to the recombinant protein (rP30) was raised in a mouse. The antibody reacted with rP30 and a 30-kDa protein of purified E. canis. Twenty-nine indirect fluorescent antibody (IFA)-positive dog plasma specimens strongly recognized the rP30 of E. canis. To evaluate whether the rP30 is a suitable antigen for serodiagnosis of canine ehrlichiosis, the immunoreactions between rP30 and the whole purified E. canis antigen were compared in the dot immunoblot assay. Dot reactions of both antigens with IFA-positive dog plasma specimens were clearly distinguishable by the naked eye from those with IFA-negative plasma specimens. By densitometry with a total of 42 IFA-positive and -negative plasma specimens, both antigens produced results similar in sensitivity and specificity. These findings suggest that the rP30 antigen provides a simple, consistent, and rapid serodiagnosis for canine ehrlichiosis. Cloning of multigenes encoding the 30-kDa major outer membrane proteins of E. canis will greatly facilitate understanding pathogenesis and immunologic study of canine ehrlichosis and provide a useful tool for phylogenetic analysis.}, } @article {pmid9695115, year = {1998}, author = {Curtis, CF and Sinkins, SP}, title = {Wolbachia as a possible means of driving genes into populations.}, journal = {Parasitology}, volume = {116 Suppl}, number = {}, pages = {S111-5}, doi = {10.1017/s0031182000084997}, pmid = {9695115}, issn = {0031-1820}, mesh = {Animals ; Crosses, Genetic ; Culicidae/genetics/*parasitology ; Host-Parasite Interactions ; Malaria/prevention & control ; Male ; Spermatocytes/parasitology ; Symbiosis ; }, abstract = {Cytoplasmic incompatibility consists of sterility in cross matings, the crossing type being maternally inherited. It can be explained by the action of Wolbachia symbionts which are transmitted through the egg cytoplasm and leave an imprint on the sperm which prevents it fertilizing unless it is 'rescued' by the action of the same type of Wolbachia in the egg. Thus matings between infected males and uninfected females are sterile, but the reciprocal matings are fertile. Hence uninfected females are at risk of failing to transmit their uninfected cytoplasm if they cross mate, but infected females are at no such risk. Therefore natural selection favours the infected state and in two wild insect populations the infection has been observed spreading. If a gene for inability to transmit malaria could be introduced into Wolbachia and if this could be introduced into Anopheles (where these symbionts appear not to occur naturally), release of a limited number of such insects should trigger a process of displacement of malaria vectors, by the non-vector type. A simple model is used to demonstrate the limitations to this process which would be introduced by immigration.}, } @article {pmid9695096, year = {1998}, author = {Dunn, AM and Rigaud, T}, title = {Horizontal transfer of parasitic sex ratio distorters between crustacean hosts.}, journal = {Parasitology}, volume = {117 (Pt 1)}, number = {}, pages = {15-19}, doi = {10.1017/s0031182098002820}, pmid = {9695096}, issn = {0031-1820}, mesh = {Adrenal Glands/ultrastructure ; Animals ; Crustacea/microbiology/*parasitology/physiology ; Female ; Feminization/parasitology/physiopathology/veterinary ; Fertility ; Fluorescent Dyes/chemistry ; Gonads/ultrastructure ; Host-Parasite Interactions ; Indoles/chemistry ; Male ; Microscopy, Electron ; Nosema/*physiology ; Ovary/parasitology/physiology ; Polymerase Chain Reaction ; Rickettsiaceae/*physiology ; Sex Ratio ; }, abstract = {Parasitic sex distorters were artificially transferred within and between crustacean host species in order to study the effects of parasitism on host fitness and sex determination and to investigate parasite host specificity. Implantation of Nosema sp. to uninfected strains of its Gammarus duebeni host resulted in an active parasite infection in the gonad of recipient females and subsequent transovarial parasite transmission. The young of artificially infected females were feminized by the parasite, demonstrating that Nosema sp. is a cause of a sex ratio distortion in its host. In contrast, we were unable to cross-infect Armadillidium vulgare with the feminizing microsporidian from G. duebeni or to cross-infect G. duebeni with the feminizing bacterium Wolbachia sp. from A. vulgare.}, } @article {pmid9684374, year = {1998}, author = {Bouchon, D and Rigaud, T and Juchault, P}, title = {Evidence for widespread Wolbachia infection in isopod crustaceans: molecular identification and host feminization.}, journal = {Proceedings. Biological sciences}, volume = {265}, number = {1401}, pages = {1081-1090}, pmid = {9684374}, issn = {0962-8452}, mesh = {Animals ; Base Sequence ; Crustacea/*microbiology ; DNA, Bacterial ; Feminization ; Molecular Sequence Data ; Phylogeny ; Rickettsiaceae/*classification/genetics ; }, abstract = {Wolbachia are maternally inherited, intracellular, alpha proteobacteria that infect a wide range of arthropods. They cause three kinds of reproductive alterations in their hosts: cytoplasmic incompatibility, parthenogenesis and feminization. There have been many studies of the distribution of Wolbachia in arthropods, but very few crustacean species are known to be infected. We investigated the prevalence of Wolbachia in 85 species from five crustacean orders. Twenty-two isopod species were found to carry these bacteria. The bacteria were found mainly in terrestrial species, suggesting that Wolbachia came from a continental environment. The evolutionary relationships between these Wolbachia strains were determined by sequencing bacterial genes and by interspecific transfers. All the bacteria associated with isopods belonged to the Wolbachia B group, based on 16S rDNA sequence data. All the terrestrial isopod symbionts in this group except one formed an independent clade. The results of interspecific transfers show evidence of specialization of Wolbachia symbionts to their isopod hosts. They also suggest that host species plays a more important role than bacterial phylogeny in determining the phenotype induced by Wolbachia infection.}, } @article {pmid9573188, year = {1998}, author = {Braig, HR and Zhou, W and Dobson, SL and O'Neill, SL}, title = {Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis.}, journal = {Journal of bacteriology}, volume = {180}, number = {9}, pages = {2373-2378}, pmid = {9573188}, issn = {0021-9193}, support = {AI 07404/AI/NIAID NIH HHS/United States ; AI 34355/AI/NIAID NIH HHS/United States ; AI40620/AI/NIAID NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Bacterial Outer Membrane Proteins/*genetics ; Base Sequence ; Cloning, Molecular ; Drosophila/*microbiology ; Escherichia coli/genetics ; *Genes, Bacterial ; Molecular Sequence Data ; Moths/microbiology ; Promoter Regions, Genetic ; Recombinant Proteins ; Rickettsiaceae/*genetics ; Sequence Homology, Amino Acid ; Symbiosis/*genetics ; }, abstract = {The maternally inherited intracellular symbiont Wolbachia pipientis is well known for inducing a variety of reproductive abnormalities in the diverse arthropod hosts it infects. It has been implicated in causing cytoplasmic incompatibility, parthenogenesis, and the feminization of genetic males in different hosts. The molecular mechanisms by which this fastidious intracellular bacterium causes these reproductive and developmental abnormalities have not yet been determined. In this paper, we report on (i) the purification of one of the most abundantly expressed Wolbachia proteins from infected Drosophila eggs and (ii) the subsequent cloning and characterization of the gene (wsp) that encodes it. The functionality of the wsp promoter region was also successfully tested in Escherichia coli. Comparison of sequences of this gene from different strains of Wolbachia revealed a high level of variability. This sequence variation correlated with the ability of certain Wolbachia strains to induce or rescue the cytoplasmic incompatibility phenotype in infected insects. As such, this gene will be a very useful tool for Wolbachia strain typing and phylogenetic analysis, as well as understanding the molecular basis of the interaction of Wolbachia with its host.}, } @article {pmid9538033, year = {1998}, author = {Schilthuizen, M and Gittenberger, E}, title = {Screening mollusks for Wolbachia infection.}, journal = {Journal of invertebrate pathology}, volume = {71}, number = {3}, pages = {268-270}, doi = {10.1006/jipa.1997.4739}, pmid = {9538033}, issn = {0022-2011}, mesh = {Animals ; Mollusca/*microbiology ; *Rickettsiaceae ; }, abstract = {We screened 38 species of mollusks for infection by Wolbachia, a bacterium that is a common endosymbiont in arthropods, where it induces alterations in reproduction. Using a PCR assay, we could not detect the symbiont in any of the samples, indicating that, in mollusks, it might be absent.}, } @article {pmid9569669, year = {1998}, author = {Zhou, W and Rousset, F and O'Neil, S}, title = {Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences.}, journal = {Proceedings. Biological sciences}, volume = {265}, number = {1395}, pages = {509-515}, pmid = {9569669}, issn = {0962-8452}, support = {AI 34355/AI/NIAID NIH HHS/United States ; AI 40620/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology ; Base Sequence ; Cloning, Molecular ; DNA Primers ; Drosophila/microbiology ; *Genes, Bacterial ; Insecta/*microbiology ; *Phylogeny ; Polymerase Chain Reaction/methods ; Rickettsiaceae/*classification/*genetics ; }, abstract = {Wolbachia are a group of intracellular inherited bacteria that infect a wide range of arthropods. They are associated with a number of different reproductive phenotypes in their hosts, such as cytoplasmic incompatibility, parthenogenesis and feminization. While it is known that the bacterial strains responsible for these different host phenotypes form a single clade within the alpha-Proteobacteria, until now it has not been possible to resolve the evolutionary relationships between different Wolbachia strains. To address this issue we have cloned and sequenced a gene encoding a surface protein of Wolbachia (wsp) from a representative sample of 28 Wolbachia strains. The sequences from this gene were highly variable and could be used to resolve the phylogenetic relationships of different Wolbachia strains. Based on the sequence of the wsp gene from different Wolbachia isolates we propose that the Wolbachia pipientis clade be initially divided into 12 groups. As more sequence information becomes available we expect the number of such groups to increase. In addition, we present a method of Wolbachia classification based on the use of group-specific wsp polymerase chain reaction (PGR) primers which will allow Wolbachia isolates to be typed without the need to clone and sequence individual Wolbachia genes. This system should facilitate future studies investigating the distribution and biology of Wolbachia strains from large samples of different host species.}, } @article {pmid9560398, year = {1998}, author = {Bordenstein, SR and Werren, JH}, title = {Effects of A and B Wolbachia and host genotype on interspecies cytoplasmic incompatibility in Nasonia.}, journal = {Genetics}, volume = {148}, number = {4}, pages = {1833-1844}, pmid = {9560398}, issn = {0016-6731}, mesh = {Animals ; Cytoplasm ; Female ; Genotype ; Male ; Reproduction/physiology ; Rickettsiaceae/*physiology ; Sexual Behavior, Animal/physiology ; Species Specificity ; Symbiosis ; Wasps/*genetics/*physiology ; }, abstract = {Wolbachia endosymbionts cause postmating reproductive isolation between the sibling species Nasonia vitripennis and N. giraulti. Most Nasonia are doubly infected with a representative from each of the two major Wolbachia groups (A and B). This study investigates the role of single (A or B) and double (A and B) Wolbachia infections in interspecies cytoplasmic incompatibility (CI) and host genomic influences on the incompatibility phenotype. Results show that the single A Wolbachia harbored in N. vitripennis (wAv) is bidirectionally incompatible with the single A Wolbachia harbored in N. giraulti (wAg). Results also indirectly show that the N. vitripennis wBv is bidirectionally incompatible with the N. giraulti wBg. The findings support current phylogenetic evidence that suggests these single infections have independent origins and were acquired via horizontal transfer. The wAv Wolbachia expresses partial CI in the N. vitripennis nuclear background. However, following genomic replacement by introgression, wAv expresses complete CI in the N. giraulti background and remains bidirectionally incompatible with wAg. Results show that double infections can reinforce interspecies reproductive isolation through the addition of incompatibility types and indicate that the host genome can influence incompatibility levels. This study has implications for host-symbiont coevolution and the role of Wolbachia in speciation.}, } @article {pmid9559095, year = {1997}, author = {Hoshizaki, S}, title = {Allozyme polymorphism and geographic variation in the small brown planthopper, Laodelphax striatellus (Homoptera: Delphacidae).}, journal = {Biochemical genetics}, volume = {35}, number = {11-12}, pages = {383-393}, doi = {10.1023/a:1022233700872}, pmid = {9559095}, issn = {0006-2928}, mesh = {Alleles ; Animals ; Gene Frequency ; Geography ; Insecta/enzymology/*genetics/*microbiology ; Isoelectric Focusing ; Japan ; *Polymorphism, Genetic ; Rickettsiaceae Infections ; Taiwan ; }, abstract = {The small brown planthopper, Laodelphax striatellus, immigrates annually into Japan over the East China Sea from the Asian mainland. It is not known whether this long-distance dispersal has any effect on the genetic structure of Japanese L. striatellus populations. The dispersal of L. striatellus is suspected to be relevant to the population dynamics of infection with the parasitic bacterium Wolbachia, which causes cytoplasmic incompatibility in L. striatellus. Wolbachia infection has spread within and among Japanese L. striatellus populations due to this cytoplasmic incompatibility. In the present study, the geographic differences among II L. striatellus populations from Japan and Taiwan was investigated using allozyme polymorphism. FST values on three enzyme loci (GPI, PGM, and AK) indicated a geographically differentiated population structure. Significant differentiation was found even among populations located along the course of the long-distance dispersal. The results indicated that long-range dispersal of L. striatellus does not occur regularly over the main islands of Japan and that it does not have a large effect on the population structure of L. striatellus. This conclusion is in agreement with the geographically variable life history of L. striatellus adapted to local climates. The short-term rice stripe epidemic, which is vectored by L. striatellus, in northwestern Kyushu, Japan, during 1985 and 1986 corresponds to these results. Based on the present findings, short-distance dispersal was considered to drive the spatial spread of Wolbachia infection among L. striatellus populations.}, } @article {pmid9535156, year = {1998}, author = {Sasaki, T and Braig, HR and O'Neill, SL}, title = {Analysis of Wolbachia protein synthesis in Drosophila in vivo.}, journal = {Insect molecular biology}, volume = {7}, number = {2}, pages = {101-105}, doi = {10.1046/j.1365-2583.1998.72057.x}, pmid = {9535156}, issn = {0962-1075}, support = {AI 34355/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/*biosynthesis ; Drosophila/metabolism/*microbiology ; Rickettsiaceae/*metabolism ; }, abstract = {Intracellular Wolbachia infections are extremely common in arthropods and exert profound control over the reproductive biology of the host. However, very little is known about the underlying molecular mechanisms which mediate these interactions with the host. We examined protein synthesis by Wolbachia in a Drosophila host in vivo by selective metabolic labelling of prokaryotic proteins and subsequent analysis by 1D and 2D gel electrophoresis. Using this method we could identify the major proteins synthesized by Wolbachia in ovaries and testes of flies. Of these proteins the most abundant was of low molecular weight and showed size variation between Wolbachia strains which correlated with the reproductive phenotype they generated in flies. Using the gel systems we employed it was not possible to identify any proteins of Wolbachia origin in the mature sperm cells of infected flies.}, } @article {pmid9523438, year = {1998}, author = {Karr, TL and Yang, W and Feder, ME}, title = {Overcoming cytoplasmic incompatibility in Drosophila.}, journal = {Proceedings. Biological sciences}, volume = {265}, number = {1394}, pages = {391-395}, pmid = {9523438}, issn = {0962-8452}, mesh = {Animals ; Drosophila/*microbiology/*physiology ; Female ; Male ; Reproduction/*physiology ; Rickettsiaceae/*physiology ; Rickettsiaceae Infections/*physiopathology ; Spermatozoa/microbiology/*physiology ; }, abstract = {The endocellular microbe Wolbachia pipientis infects a wide variety of invertebrate species, in which its presence is closely linked to a form of reproductive failure termed cytoplasmic incompatibility (CI). CI renders infected males unable to father offspring when mated to uninfected females. Because CI can dramatically affect fitness in natural populations, mechanisms that abate CI can have equally large impacts on fitness. We have discovered that repeated copulation by Wolbachia-infected male Drosophila simulans significantly diminishes CI. Repeated copulation does not prevent Wolbachia from populating developing spermatids, but may reduce the time during spermatogenesis when Wolbachia can express CI. This restoration of fertility in premated infected males could have important implications for Wolbachia transmission and persistence in nature and for its exploitation as an agent of biological pest control.}, } @article {pmid9511829, year = {1998}, author = {Popov, VL and Han, VC and Chen, SM and Dumler, JS and Feng, HM and Andreadis, TG and Tesh, RB and Walker, DH}, title = {Ultrastructural differentiation of the genogroups in the genus Ehrlichia.}, journal = {Journal of medical microbiology}, volume = {47}, number = {3}, pages = {235-251}, doi = {10.1099/00222615-47-3-235}, pmid = {9511829}, issn = {0022-2615}, support = {AI 31431/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cells, Cultured ; Ehrlichia/classification/genetics/*ultrastructure ; Ehrlichia chaffeensis/ultrastructure ; Horses ; Humans ; Macrophages/microbiology ; Mice ; Mice, Inbred BALB C ; RNA, Ribosomal, 16S/analysis ; }, abstract = {Ultrastructural characteristics of 15 strains and isolates of ehrlichiae belonging to three genogroups, or clades of genetically related organisms united in the genera Ehrlichia, Cowdria, Anaplasma, Neorickettsia and a strain of Wolbachia pipientis which represents a fourth genogroup in this cluster of species, were studied in continuous cell culture or in vivo: E. canis (Oklahoma strain and VHE isolate), E. muris (AS 145), E. chaffeensis (Arkansas, 91HE17 and Sapulpa), human granulocytic ehrlichiae (HGE)(BDS, 96HE27, 96HE37, #54, #55 and #72), E. equi (MRK), E. sennetsu (Miyayama), E. risticii (HRC-IL). Wolbachia pipientis was studied in the naturally infected Aedes albopictus mosquito cell line Aa23. All organisms were similar in the normal ultrastructure of individual cells and in the ability to form abnormal, pathological ehrlichial cells of the same type irrespective of the species. Normally all ehrlichiae studied in cell culture existed in two morphological forms - reticulate and dense-cored cells, both of which could divide by binary fission. Most alterations were related to their membranes, especially the cell wall. Differences in the structure of intravacuolar microcolonies (morulae) of ehrlichiae and their inter-relations with the host cells allowed differentiation of the genogroups: the E. canis-E. chaffeensis-E. muris genogroup formed large morulae, with many ehrlichiae, often suspended in a fibrillar matrix, and the host cell mitochondria and endoplasmic reticulum usually aggregated near the morulae and were in contact with the morula membrane; the E. phagocytophila-E. equi-HGE group morulae had no fibrillar matrix, no contacts with host cell mitochodria, and they did not aggregate around the morulae; E. sennetsu-E. risticii group usually developed in small individual vacuoles that did not fuse with each other and divided along with the ehrlichiae.}, } @article {pmid9495337, year = {1998}, author = {Bourtzis, K and Dobson, SL and Braig, HR and O'Neill, SL}, title = {Rescuing Wolbachia have been overlooked.}, journal = {Nature}, volume = {391}, number = {6670}, pages = {852-853}, doi = {10.1038/36017}, pmid = {9495337}, issn = {0028-0836}, mesh = {Animals ; Crosses, Genetic ; Drosophila/genetics/*microbiology ; Drosophila melanogaster/genetics/microbiology ; Female ; Male ; Ovary/microbiology ; Reproduction ; Rickettsiaceae/genetics/*physiology ; Species Specificity ; Spermatozoa/microbiology ; Testis/microbiology ; }, } @article {pmid9405738, year = {1998}, author = {Zchori-Fein, E and Roush, RT and Rosen, D}, title = {Distribution of parthenogenesis-inducing symbionts in ovaries and eggs of Aphytis (Hymentoptera: Aphelinidae).}, journal = {Current microbiology}, volume = {36}, number = {1}, pages = {1-8}, doi = {10.1007/s002849900270}, pmid = {9405738}, issn = {0343-8651}, mesh = {Animals ; Bacteria/*isolation & purification ; Female ; Hymenoptera/*microbiology ; Microscopy, Confocal ; Microscopy, Electron ; Ovary/microbiology/ultrastructure ; Ovum/microbiology ; Symbiosis ; }, abstract = {In parasitic Hymenoptera, severe sex ratio distortions are sometimes associated with the presence of symbiotic microorganisms. Symbiont-induced uniparental reproduction is found in several species of the genus Aphytis and is suspected in others. These intracellular bacteria (called Wolbachia) reside within the egg cytoplasm. In order to follow their vertical transmission, the distribution pattern of the symbionts in ovaries and eggs of two uniparental lines (A. lingnanensis and A. yanonensis) was studied by electron, confocal, and two-photon microscopy. The results obtained suggest that Wolbachia move from nurse cells to developing oocytes. In freshly laid eggs, the symbionts are concentrated in the posterior pole, away from the micropyle, and in later stages of embryogenesis they are found surrounding nuclei throughout the embryo.}, } @article {pmid9475734, year = {1998}, author = {Hoffmann, AA and Hercus, M and Dagher, H}, title = {Population dynamics of the Wolbachia infection causing cytoplasmic incompatibility in Drosophila melanogaster.}, journal = {Genetics}, volume = {148}, number = {1}, pages = {221-231}, pmid = {9475734}, issn = {0016-6731}, mesh = {Analysis of Variance ; Animals ; Animals, Laboratory ; Breeding ; Cytoplasm/*physiology ; Drosophila melanogaster/growth & development/*microbiology ; Female ; Male ; Rickettsiaceae ; Rickettsiaceae Infections/*physiopathology/transmission ; Spermatozoa/physiology ; }, abstract = {Field populations of Drosophila melanogaster are often infected with Wolbachia, a vertically transmitted microorganism. Under laboratory conditions the infection causes partial incompatibility in crosses between infected males and uninfected females. Here we examine factors influencing the distribution of the infection in natural populations. We show that the level of incompatibility under field conditions was much weaker than in the laboratory. The infection was not transmitted with complete fidelity under field conditions, while field males did not transmit the infection to uninfected females and Wolbachia did not influence sperm competition. There was no association between field fitness as measured by fluctuating asymmetry and the infection status of adults. Infected field females were smaller than uninfecteds in some collections from a subtropical location, but not in other collections from the same location. Laboratory cage studies showed that the infection did not change in frequency when populations were maintained at a low larval density, but it decreased in frequency at a high larval density. Monitoring of infection frequencies in natural populations indicated stable frequencies in some populations but marked fluctuations in others. Simple models suggest that the infection probably provides a fitness benefit for the host in order to persist in populations. The exact nature of this benefit remains elusive.}, } @article {pmid9327558, year = {1997}, author = {Niebylski, ML and Peacock, MG and Fischer, ER and Porcella, SF and Schwan, TG}, title = {Characterization of an endosymbiont infecting wood ticks, Dermacentor andersoni, as a member of the genus Francisella.}, journal = {Applied and environmental microbiology}, volume = {63}, number = {10}, pages = {3933-3940}, pmid = {9327558}, issn = {0099-2240}, mesh = {Animals ; Arachnid Vectors/*microbiology/ultrastructure ; Base Sequence ; Chlorocebus aethiops ; DNA Primers/genetics ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Dermacentor/*microbiology/ultrastructure ; Female ; Francisella/genetics/*isolation & purification/ultrastructure ; Microscopy, Electron ; Ovary/microbiology ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Symbiosis ; Vero Cells ; }, abstract = {A microorganism (Dermacantor andersoni symbiont [DAS]) infecting Rocky Mountain wood ticks (D. andersoni) collected in the Bitterroot Mountains of western Montana was characterized as an endosymbiont belonging to the genus Francisella. Previously described as Wolbachia like, the organism's DNA was amplified from both naturally infected tick ovarial tissues and Vero cell cultures by PCR assay with primer sets derived from eubacterial 16S ribosomal DNA (rDNA) and Francisella membrane protein genes. The 16S rDNA gene sequence of the DAS was most similar (95.4%) to that of Francisella tularensis subsp. tularensis. Through a combination of Giménez staining, PCR assay, and restriction fragment length polymorphism analysis, 102 of 108 female ticks collected from 1992 to 1996 were infected. Transovarial transmission to female progeny was 95.6%, but we found no evidence of horizontal transmission.}, } @article {pmid9327557, year = {1997}, author = {Noda, H and Munderloh, UG and Kurtti, TJ}, title = {Endosymbionts of ticks and their relationship to Wolbachia spp. and tick-borne pathogens of humans and animals.}, journal = {Applied and environmental microbiology}, volume = {63}, number = {10}, pages = {3926-3932}, pmid = {9327557}, issn = {0099-2240}, support = {AR37909/AR/NIAMS NIH HHS/United States ; }, mesh = {Animals ; Arachnid Vectors/*microbiology ; Bacteria/classification/genetics/*isolation & purification ; Base Sequence ; DNA Primers/genetics ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Dogs ; Humans ; Ixodes/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/classification/genetics/*isolation & purification ; Symbiosis ; Ticks/*microbiology ; }, abstract = {The presence, internal distribution, and phylogenetic position of endosymbiotic bacteria from four species of specific-pathogen-free ticks were studied. These included the hard ticks Ixodes scapularis (the black-legged tick), Rhipicephalus sanguineus (the brown dog tick), and Haemaphysalis longicornis and the African soft tick Ornithodoros moubata. PCR assays for bacteria, using two sets of general primers for eubacterial 16S and 23S rRNA genes (rDNAs) and seven sets of specific primers for wolbachial, rickettsial, or Francisella genes, indicated that I. scapularis possessed symbiotic rickettsiae in the ovaries and that the other species harbored eubacteria in both the ovaries and Malpighian tubules. Phylogenetic analysis based on the sequence of 16S rDNA indicated that the symbiont of I. scapularis belonged to the alpha subgroup of proteobacteria and was closely related to the members of the genus Rickettsia. The other species had similar microorganisms in the ovaries and Malpighian tubules, which belonged to the gamma subgroup of proteobacteria, and formed a monophyletic group with the Q-fever pathogen, Coxiella burnetii. O. moubata harbored another symbiont, which formed a monophyletic group with Francisella tularensis and Wolbachia persica, the latter a symbiont previously isolated from Malpighian tubules of the soft tick Argas (Persicargas) arboreus. Thus, the symbionts of these four tick species were not related to the Wolbachia species found in insects. The two symbionts that live in the Malpighian tubules, one closely related to C. burnetii and the other closely related to F. tularensis, appear to be of ancient origin and be widely distributed in ticks.}, } @article {pmid9326628, year = {1997}, author = {Giordano, R and Jackson, JJ and Robertson, HM}, title = {The role of Wolbachia bacteria in reproductive incompatibilities and hybrid zones of Diabrotica beetles and Gryllus crickets.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {94}, number = {21}, pages = {11439-11444}, pmid = {9326628}, issn = {0027-8424}, mesh = {Animals ; Bacterial Proteins/*genetics ; *Biological Evolution ; Coleoptera/genetics/*microbiology/*physiology ; Crosses, Genetic ; *Cytoskeletal Proteins ; Evolution, Molecular ; Female ; GTP-Binding Proteins/genetics ; Geography ; Male ; Molecular Sequence Data ; Orthoptera/genetics/microbiology/*physiology ; Phylogeny ; Reproduction/genetics ; Rickettsia/genetics/*physiology ; United States ; Zea mays/parasitology ; }, abstract = {A rickettsial bacterium in the genus Wolbachia is the cause of a unidirectional reproductive incompatibility observed between two major beetle pests of maize, the western corn rootworm, Diabrotica virgifera virgifera, and the Mexican corn rootworm, D. v. zeae. These subspecies are allopatric except for two known regions of sympatry in Texas and Mexico. We demonstrate that populations of D. v. virgifera, with the exception of two populations in southern Arizona, are infected with a strain of Wolbachia. Populations of D. v. zeae are not infected. Treatment of D. v. virgifera with tetracycline eliminated the Wolbachia and removed the reproductive incompatibility. Similar patterns of reproductive incompatibility exist among taxa of the cricket genus Gryllus. Gryllus assimilis, G. integer, G. ovisopis, G. pennsylvanicus, and G. rubens are infected with Wolbachia whereas G. firmus is usually not. Populations of G. rubens and G. ovisopis carry the same Wolbachia strain, which is distinct from that of G. integer. G. pennsylvanicus is infected with two Wolbachia strains, that found in G. rubens and one unique to G. pennsylvanicus. Moreover, a proportion of G. pennsylvanicus individuals harbors both strains. Wolbachia may have influenced speciation in some members of the genus Gryllus by affecting the degree of hybridization between species. Given that Wolbachia infections are relatively common in insects, it is likely that other insect hybrid zones may be influenced by infections with Wolbachia.}, } @article {pmid9326576, year = {1997}, author = {Werren, JH}, title = {Wolbachia run amok.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {94}, number = {21}, pages = {11154-11155}, pmid = {9326576}, issn = {0027-8424}, mesh = {Animals ; Aphids/microbiology ; Drosophila melanogaster/microbiology ; Gram-Negative Bacteria/*physiology ; Humans ; Invertebrates ; Oocytes/microbiology ; Rickettsia/*physiology ; Rickettsia Infections/*transmission ; Symbiosis ; }, } @article {pmid9380712, year = {1997}, author = {Min, KT and Benzer, S}, title = {Wolbachia, normally a symbiont of Drosophila, can be virulent, causing degeneration and early death.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {94}, number = {20}, pages = {10792-10796}, pmid = {9380712}, issn = {0027-8424}, support = {AG 12289/AG/NIA NIH HHS/United States ; EY 09278/EY/NEI NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/genetics/*microbiology/ultrastructure ; Microscopy, Electron ; Phenotype ; Rickettsia/*pathogenicity/ultrastructure ; }, abstract = {Wolbachia, a maternally transmitted microorganism of the Rickettsial family, is known to cause cytoplasmic incompatibility, parthenogenesis, or feminization in various insect species. The bacterium-host relationship is usually symbiotic: incompatibility between infected males and uninfected females can enhance reproductive isolation and evolution, whereas the other mechanisms enhance progeny production. We have discovered a variant Wolbachia carried by Drosophila melanogaster in which this cozy relationship is abrogated. Although quiescent during the fly's development, it begins massive proliferation in the adult, causing widespread degeneration of tissues, including brain, retina, and muscle, culminating in early death. Tetracycline treatment of carrier flies eliminates both the bacteria and the degeneration, restoring normal life-span. The 16s rDNA sequence is over 98% identical to Wolbachia known from other insects. Examination of laboratory strains of D. melanogaster commonly used in genetic experiments reveals that a large proportion actually carry Wolbachia in a nonvirulent form, which might affect their longevity and behavior.}, } @article {pmid9401467, year = {1997}, author = {Masui, S and Sasaki, T and Ishikawa, H}, title = {groE-homologous operon of Wolbachia, an intracellular symbiont of arthropods: a new approach for their phylogeny.}, journal = {Zoological science}, volume = {14}, number = {4}, pages = {701-706}, doi = {10.2108/zsj.14.701}, pmid = {9401467}, issn = {0289-0003}, mesh = {Amino Acid Sequence ; Animals ; Arthropods/*microbiology ; Bacterial Proteins/*genetics ; Base Sequence ; Chaperonins ; Escherichia coli Proteins ; Heat-Shock Proteins/*genetics ; Molecular Sequence Data ; *Operon ; Phylogeny ; Rickettsia/classification/*genetics ; }, abstract = {Wolbachia, a member of rickettsia found in the cells of many arthropod species, are cytoplasmically inherited bacteria which interfere with host's sexuality and reproduction. Wolbachia strains have been phylogenetically divided into A and B groups based on the nucleotide sequences of their ftsZ genes. In an attempt to further define the phylogenetical relationship among these endosymbionts, we cloned and sequenced the entire length of the groE operon of a Wolbachia harbored by a cricket. The operon encoded two heat shock proteins, which represented the third and fourth proteins of any Wolbachia ever characterized. Also, 800 bp stretches of the groE operons of several other Wolbachia were sequenced, and a phylogenetic tree was constructed based on the results. The groE tree defined the relationship among A group Wolbachia strains that had not been successfully resolved by the ftsZ tree, and suggested unexpected horizontal transmission of these bacteria.}, } @article {pmid9272448, year = {1997}, author = {Hypsa, V and Aksoy, S}, title = {Phylogenetic characterization of two transovarially transmitted endosymbionts of the bedbug Cimex lectularius (Heteroptera:Cimicidae).}, journal = {Insect molecular biology}, volume = {6}, number = {3}, pages = {301-304}, doi = {10.1046/j.1365-2583.1997.00178.x}, pmid = {9272448}, issn = {0962-1075}, support = {AI-34033/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteria/*genetics ; Bedbugs/*microbiology ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Female ; Molecular Sequence Data ; Ovary/microbiology ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsieae/*genetics ; *Symbiosis ; }, abstract = {Two different inherited bacterial symbionts from ovary tissue of the bedbug Cimex lectularius were characterized by gene amplification and sequencing analysis of their 16S rDNA gene. The first bacterium belongs to the Wolbachia subgroup of the alpha-Proteobacteria, the second is a member of the gamma-subdivision, and is closely related to the bacterial parasite of the leafhopper Euscelidius variegatus (BEV) which was shown to be capable of transovarial transmission. The high similarity (> 97%) between the C. lectularius symbiont and BEV indicates that these two microorganisms belong to the same lineage and share the ability to invade distant insect hosts to fulfill their symbiotic functions and to establish transovarial transmission to future generations.}, } @article {pmid9263471, year = {1997}, author = {Fialho, RF and Stevens, L}, title = {Molecular evidence for single Wolbachia infections among geographic strains of the flour beetle Tribolium confusum.}, journal = {Proceedings. Biological sciences}, volume = {264}, number = {1384}, pages = {1065-1068}, pmid = {9263471}, issn = {0962-8452}, mesh = {Animals ; Base Sequence ; Crosses, Genetic ; DNA Primers ; Female ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Reproduction ; Rickettsia/*classification/genetics/*isolation & purification ; Rickettsia Infections ; Tribolium/*microbiology ; }, abstract = {Infections with the rickettsial microorganism Wolbachia are cytoplasmically inherited and occur in a wide range of insect species and several other arthropods. Wolbachia infection often results in unidirectional cytoplasmic incompatibility (CI): crosses between infected males and uninfected females are incompatible and show a reduction of progeny or complete inviability. Unidirectional CI can also occur when males harbouring two incompatible Wolbachia strains are crossed with females infected with only one of the two strains. In the flour beetle Tribolium confusum, Wolbachia infections are of particular interest because of the severity of incompatibility. Typically, no progeny results from the incompatible cross, whereas only partial incompatibility is observed in most other hosts. Werren et al. (1995a) reported that Wolbachia infections in T. confusum consist of two bacterial strains belonging to distinct phylogenic groups, based on PCR amplification and sequence analysis of the bacterial cell division gene ftsZ. However, Fialho & Stevens (1996) showed that eight strains of T. confusum were infected with a single and common incompatibility type. Here we report analysis of the ftsZ gene by specific PCR amplification. Diagnostic restriction enzyme assays revealed no evidence of double infections in 11 geographic strains of T. confusum, including the strain examined by Werren et al. (1995a). Further, sequence analysis of the Wolbachia ftsZ gene and an internal transcribed spacer (ITS) region in two of these strains displayed no nucleotide variation or evidence of polymorphisms. Results suggest that T. confusum is infected with B-group Wolbachia only.}, } @article {pmid9099582, year = {1997}, author = {Aksoy, S and Chen, X and Hypsa, V}, title = {Phylogeny and potential transmission routes of midgut-associated endosymbionts of tsetse (Diptera:Glossinidae).}, journal = {Insect molecular biology}, volume = {6}, number = {2}, pages = {183-190}, doi = {10.1111/j.1365-2583.1997.tb00086.x}, pmid = {9099582}, issn = {0962-1075}, support = {AI-34033/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Bacteria/*classification/*genetics ; Base Sequence ; DNA ; Digestive System/microbiology ; Molecular Sequence Data ; Phylogeny ; *Symbiosis ; Tsetse Flies/*microbiology ; }, abstract = {Many tsetse species (Diptera: Glossinidae) harbour two morphologically different intracellular endosymbiotic microorganisms associated with gut tissue: primary (P) and secondary (S) endosymbionts. The P-endosymbionts of tsetse (Wigglesworthia glossinidia) are sequestered in specialized epithelial cells, bacteriocytes, which form a structure (bacteriome) in the anterior portion of the gut. Phylogenetic characterization of P-endsymbionts from the three subgenera of genus Glossina has shown that these organisms constitute a distinct lineage within the gamma-subdivision of Proteobacteria and have evolved concordantly with their insect host species, suggesting an evolutionarily ancient association for this symbiosis. The S-endosymbiont is a smaller (1-2 micron) gram-negative rod and is harboured in midgut epithelial cells. Its phylogenetic characterization from Glossina morsitans morsitans had shown that it is a member of the family Enterobacteriaceae within the gamma-3 subdivision of the Proteobacteria, closely related to enteric bacteria. Some tsetse species harbour a third bacterium in their reproductive tissue, which was shown phylogenetically to belong to to the Wolbachia pipientis assemblage of microorganisms. Here, we show that S-endosymbionts from five tsetse species, representing all three subgenera, form a cluster of closely related microorganisms, based on their almost identical 16S rRNA gene sequences. The high similarity provides strong evidence of recent independent acquisition of S-endosymbionts by individual tsetse species, unlike Wigglesworthia which displays concordant evolution with host insect species. A PCR-based assay and restriction fragment length polymorphism (RFLP) analysis was developed to localize the S-endosymbionts and Wigglesworthia in ovary, egg, milk-gland and spermatheca tissues in order to investigate the potential routes for the vertical transmission of these symbionts to the intrauterine larvae. Only S-endosymbionts were found to infect milk gland tissue, suggesting that milk gland secretions represent a route of transmission for these symbionts into the developing larva. The ovary tissue was found to harbour only Wolbachia, confirming its transovarial transmission, whereas the mode of transmission of Wigglesworthia remains unknown.}, } @article {pmid9107051, year = {1997}, author = {Schilthuizen, M and Stouthamer, R}, title = {Horizontal transmission of parthenogenesis-inducing microbes in Trichogramma wasps.}, journal = {Proceedings. Biological sciences}, volume = {264}, number = {1380}, pages = {361-366}, pmid = {9107051}, issn = {0962-8452}, mesh = {Alphaproteobacteria/classification/*genetics/*physiology ; Animals ; Anti-Bacterial Agents/pharmacology ; DNA Primers ; DNA, Bacterial/isolation & purification ; Female ; Lepidoptera/parasitology ; Ovum ; *Parthenogenesis ; *Phylogeny ; Polymerase Chain Reaction ; Reproduction/drug effects ; Symbiosis ; Wasps/*microbiology/*physiology ; }, abstract = {Complete parthenogenesis (thelytoky) in species of the parasitic wasp Trichogramma is usually caused by the cytoplasmically inherited bacterium Wolbachia. This symbiont induces gamete duplication, which, in these haplodiploid organisms, results in all-female broods. Antibiotic treatment "cures' this condition, restoring normal sexual reproduction. Phylogenetic analysis of Wolbachia has shown that, in contrast with the strains in other host organisms (where the symbiont also induces different reproductive alterations), those in Trichogramma form a monophyletic group. This might be an indication of symbiont-host cocladogenesis. To test this, we performed comparative molecular phylogenetics on 20 parthenogenetic Trichogramma cultures and their Wolbachiae. We conclude that there is, in fact, little evidence for cocladogenesis. Instead, the phylogenetic distribution of the symbionts appears to result from occasional horizontal transmission, which probably takes place inside the hosts of Trichogramma parasitoids (usually lepidopteran eggs). This study therefore suggests that parthenogenesis is not only curable, it can sometimes be contagious also.}, } @article {pmid9419851, year = {1997}, author = {Bandi, C and Sironi, M and Nalepa, CA and Corona, S and Sacchi, L}, title = {Phylogenetically distant intracellular symbionts in termites.}, journal = {Parassitologia}, volume = {39}, number = {1}, pages = {71-75}, pmid = {9419851}, issn = {0048-2951}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Cockroaches/microbiology ; Fat Body/*microbiology ; Insecta/*microbiology ; Microscopy, Electron ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Species Specificity ; Symbiosis ; }, abstract = {Cockroaches are known to harbour intracellular bacteria in specialised cells (mycetocytes, or bacteriocytes) of the fat body. In termites, mycetocyte bacteria have been observed only in Mastotermes darwiniensis. These symbionts are thought to have originated from a bacterium that infected an ancestor common to cockroaches and termites. Thus, loss of the infection should have occurred during evolution in all termite lineages, with the exception of that leading to M. darwiniensis. One might suspect that traces of the ancient infection may be present in some termites, in the form of non-mycetocyte intracellular bacteria (e.g. a small number of bacteria within normal cells). Indeed, circumstantial evidence for the presence of intracellular bacteria in two termite species has been reported. However, no data are available on the actual distribution of these bacteria in termites, or on their relationships with the mycetocyte bacteria of cockroaches and M. darwiniensis. In this paper we report results indicating that non-mycetocyte intracellular bacteria are widespread in termites. These results were obtained by electron microscopy on representatives of nine termite species. In addition, sequence analysis of the 16S rRNA genes indicated that the non-mycetocyte bacteria of termites belong to the wolbachia group of the alpha-2 subclass of the proteobacteria. These latter bacteria are not related to the mycetocyte bacteria of cockroaches and M. darwiniensis, which belong to the blattabacterium group of the flavobacteria-bacteroides. PCR analyses with primers specific for wolbachia or blattabacterium provided further support for the identification of the observed non-mycetocyte bacteria as members of the wolbachia group.}, } @article {pmid9056458, year = {1997}, author = {Bjørnson, S and Steiner, MY and Keddie, BA}, title = {Birefringent Crystals and Abdominal Discoloration in the Predatory Mite Phytoseiulus persimilis (Acari: Phytoseiidae).}, journal = {Journal of invertebrate pathology}, volume = {69}, number = {2}, pages = {85-91}, doi = {10.1006/jipa.1996.4627}, pmid = {9056458}, issn = {1096-0805}, abstract = {In response to grower complaints of poor performance of Phytoseiulus persimilis, mites from 14 commercial insectaries and research colonies were examined for pathogens. Some were found to have abdominal discolorations, manifested initially as two white stripes along the dorsal sides of the body within the Malpighian tubules. Advanced signs appeared as a large, centrally located, white spot or U-shaped discoloration in the distal opisthosoma within the rectum/anal atrium. White material often accumulated and hardened within the anus and formed a rectal plug that inhibited further excretion. Most affected mites were lethargic. Adults and immatures with abdominal discoloration contained numerous densely packed, birefringent, dumbbell-shaped entities. Though occasionally observed in the colon, they occurred most frequently within the Malpighian tubules and/or rectum and anal atrium. Dumbbells measured 2-4 &mgr;m long and contained prominent concentric rings. When observed by transmission electron microscopy, the entities lacked cellular organelles. Asymptomatic mites contained few or no such entities. Dumbbell-shaped inclusions were observed in P. persimilis from all sources examined. High levels of potassium, low levels of phosphorous and sulfur, and traces of chlorine were detected by energy-dispersive X-ray analysis. Guanine and uric acid, known nitrogenous wastes of arachnids, do not contain these elements. The chemical composition and structure indicate that the dumbbells are crystals. Both asymptomatic mites and those specimens exhibiting abdominal discoloration were examined for pathogens using light and transmission electron microscopy. Microsporidia, virus-like particles, and a rickettsia (genus Wolbachia) were observed in some mites but showed no correlation with white abdominal discoloration or associated crystal formation. Neither were pathogens always detected in symptomatic mites. Although birefringent crystals may be naturally occurring excretory products, the cause of white abdominal signs associated with crystal accumulation in P. persimilis is unknown. These signs indicate overall poor health.}, } @article {pmid9061971, year = {1997}, author = {Guillemaud, T and Pasteur, N and Rousset, F}, title = {Contrasting levels of variability between cytoplasmic genomes and incompatibility types in the mosquito Culex pipiens.}, journal = {Proceedings. Biological sciences}, volume = {264}, number = {1379}, pages = {245-251}, pmid = {9061971}, issn = {0962-8452}, mesh = {Animals ; Culex/*genetics ; DNA, Mitochondrial/*genetics ; Molecular Sequence Data ; }, abstract = {Reproductive incompatibilities called cytoplasmic incompatibilities are known to affect a large number of arthropod species and are mediated by Wolbachia, a maternally transmitted microorganism. The crossing relationships between strains of potential hosts define their incompatibility types and it is generally assumed that differences between strains of Wolbachia induce different crossing types. Among all the described host species, the mosquito, Culex pipiens, displays the greatest variability of cytoplasmic incompatibility crossing types. We analysed mitochondrial and bacterial DNA variability in Culex pipiens in order to investigate some possible causes of incompatibility crossing type variability. We sequenced fragments of the ftsZ gene, and the A + T-rich control region of the mtDNA. We also sequenced the second subunit of the mitochondrial cytochrome oxidase (COII) gene, in Culex pipiens and a closely related species, C. torrentium, in order to verify the usefulness of the A + T-rich region for the present purposes. No variability was found in the Wolbachia ftsZ gene fragment, and very limited variation of the mitochondrial marker whatever the compatibility type or the origin of the host. A low variability was found in the A + T-rich region and comparison of divergence of the A + T-rich region and COII gene between C. pipiens and C. torrentium did not reveal any special constraints affecting this region. In contrast to observations in other host species, variability of incompatibility crossing types is not due to multiple infections by distantly related Wolbachia strains.}, } @article {pmid9013253, year = {1997}, author = {O'Neill, SL and Pettigrew, MM and Sinkins, SP and Braig, HR and Andreadis, TG and Tesh, RB}, title = {In vitro cultivation of Wolbachia pipientis in an Aedes albopictus cell line.}, journal = {Insect molecular biology}, volume = {6}, number = {1}, pages = {33-39}, doi = {10.1046/j.1365-2583.1997.00157.x}, pmid = {9013253}, issn = {0962-1075}, support = {AI28778/AI/NIAID NIH HHS/United States ; AI34355/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*cytology ; Animals ; Cell Line ; Rickettsiaceae/*growth & development ; }, abstract = {A continuous cell line, Aa23, was established from eggs of a strain of the Asian tiger mosquito, Aedes albopictus, naturally infected with the intracellular symbiont Wolbachia pipientis. The resulting cell line was shown to be persistently infected with the bacterial endosymbiont. Treatment with antibiotics cured the cells of the infection. In the course of establishing this cell line it was noticed that RFLPs in the PCR products of two Wolbachia genes from the parental mosquitoes were fixed in the infected cell line. This indicates that the mosquito host was naturally superinfected with different Wolbachia strains, whereas the infected cell line derived from these mosquitoes only contained one of the original Wolbachia strains. The development of an in vitro culture system for this fastidious microorganism should facilitate molecular analysis of the reproduction distorting phenotypes it induces in natural arthropod hosts.}, } @article {pmid9044057, year = {1997}, author = {Callaini, G and Dallai, R and Riparbelli, MG}, title = {Wolbachia-induced delay of paternal chromatin condensation does not prevent maternal chromosomes from entering anaphase in incompatible crosses of Drosophila simulans.}, journal = {Journal of cell science}, volume = {110 (Pt 2)}, number = {}, pages = {271-280}, doi = {10.1242/jcs.110.2.271}, pmid = {9044057}, issn = {0021-9533}, mesh = {Anaphase ; Animals ; Chromatin ; Chromosomes ; Crosses, Genetic ; Drosophila/*embryology ; Female ; Male ; Meiosis/physiology ; Oocytes/physiology ; Rickettsia/*physiology ; Sperm-Ovum Interactions/physiology ; Spermatozoa/physiology ; Spindle Apparatus/physiology ; Zygote/*physiology ; }, abstract = {The behavior of parental chromosomes during the first mitosis of Drosophila simulans zygotes obtained from unidirectional incompatible crosses is described and it is demonstrated that the condensation of parental chromatin complements was asynchronous. The timing of paternal chromatin condensation appeared to be delayed in these embryos, so that condensed maternal chromosomes and entangled prophase-like paternal fibers congressed in the equatorial plane of the first metaphase spindle. At anaphase the maternal chromosomes migrated to opposite poles of the spindle, whereas the paternal chromatin lagged in the midzone of the spindle. This resulted in dramatic errors in paternal chromatin inheritance leading to the formation of embryos with aneuploid or haploid nuclei. These observations suggest that the anaphase onset of maternal chromosomes is unaffected by the improper alignment of the paternal complement. Since the first metaphase spindle of the Drosophila zygote consists of twin bundles of microtubules each holding one parental complement, we suspect that each half spindle regulates the timing of anaphase onset of its own chromosome set. In normal developing embryos, the fidelity of chromosome transmission is presumably ensured by the relative timing required to prepare parental complements for the orderly segregation that occurs during the metaphase-anaphase transition.}, } @article {pmid8978041, year = {1996}, author = {Ballard, JW and Hatzidakis, J and Karr, TL and Kreitman, M}, title = {Reduced variation in Drosophila simulans mitochondrial DNA.}, journal = {Genetics}, volume = {144}, number = {4}, pages = {1519-1528}, pmid = {8978041}, issn = {0016-6731}, support = {GM-39355/GM/NIGMS NIH HHS/United States ; GM-47962/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; DNA, Mitochondrial/*genetics ; Drosophila/*genetics/microbiology ; Evolution, Molecular ; *Genetic Variation ; Rickettsia Infections/*genetics ; }, abstract = {We investigated the evolutionary dynamics of infection of a Drosophila simulans population by a maternally inherited insect bacterial parasite, Wolbachia, by analyzing nucleotide variability in three regions of the mitochondrial genome in four infected and 35 uninfected lines. Mitochondrial variability is significantly reduced compared to a noncoding region of a nuclear-encoded gene in both uninfected and pooled samples of flies, indicating a sweep of genetic variation. The selective sweep of mitochondrial DNA may have been generated by the fixation of an advantageous mitochondrial gene mutation in the mitochondrial genome. Alternatively, the dramatic reduction in mitochondrial diversity may be related to Wolbachia.}, } @article {pmid8913750, year = {1996}, author = {Bourtzis, K and Nirgianaki, A and Markakis, G and Savakis, C}, title = {Wolbachia infection and cytoplasmic incompatibility in Drosophila species.}, journal = {Genetics}, volume = {144}, number = {3}, pages = {1063-1073}, pmid = {8913750}, issn = {0016-6731}, mesh = {Animals ; Bacterial Proteins/genetics ; DNA-Binding Proteins/genetics ; Drosophila/genetics/*microbiology/*physiology ; Female ; *Fertility ; Male ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; Rickettsia/genetics/*isolation & purification ; }, abstract = {Forty-one stocks from 30 Drosophila species were surveyed for Wolbachia infection using PCR technology. D. sechellia and two strains of D. auraria were found to be infected and were tested for the expression of cytoplasmic incompatibility, along with D. ananassae and D. melanogaster strains, which are already known to be infected. D. ananassae and D. melanogaster show levels of incompatibility up to 25%, while D. auraria and D. sechellia exhibit levels of egg mortality approximately 60%. A dot-blot assay using the dnaA sequence as probe was developed to assess the infection levels in individual males that were used in incompatibility crosses. A positive correlation between bacterial density and cytoplasmic incompatibility was observed. The stocks examined can be clustered into at least two groups, depending on the levels of infection relative to the degree of cytoplasmic incompatibility exhibited. One group, containing D. simulans Hawaii, D. sechellia, and D. auraria, exhibits high levels of cytoplasmic incompatibility relative to levels of infection; all the other species and D. simulans Riverside exhibit significantly lower levels of cytoplasmic incompatibility relative to levels of infection. These data show that, in addition to bacterial density, bacterial and/ or host factors also affect the expression of cytoplasmic incompatibility.}, } @article {pmid8856963, year = {1996}, author = {Breeuwer, JA and Jacobs, G}, title = {Wolbachia: intracellular manipulators of mite reproduction.}, journal = {Experimental & applied acarology}, volume = {20}, number = {8}, pages = {421-434}, pmid = {8856963}, issn = {0168-8162}, mesh = {Animals ; Bacterial Proteins/*genetics ; *Cytoskeletal Proteins ; DNA, Bacterial/*analysis ; Female ; Mites/*microbiology ; Polymerase Chain Reaction ; Rickettsiaceae/genetics/*isolation & purification ; }, abstract = {Cytoplasmically transmitted Wolbachia (alpha-Proteobacteria) are a group of closely related intracellular microorganisms that alter reproduction in arthropods. They are found in a few isopods and are widespread in insects. Wolbachia are implicated as the cause of parthenogenesis in parasitic wasps, feminization in isopods and reproductive (cytoplasmic) incompatibility in many insects. Here we report on the widespread occurrence of Wolbachia in spider mites and predatory mites based on a PCR assay for a 730 bp fragment of the ftsZ gene with primers that are specific for Wolbachia. An additional PCR, using two primer pairs that amplify a 259 bp region of the ftsZ gene that are diagnostic for the two Wolbachia subdivisions A and B, showed that infected mites only carried type B and not type A Wolbachia. The fact that some species tested negative for Wolbachia does not mean that the entire species is uninfected. We found that natural populations of Tetranychus urticae are polymorphic for the infection. The possible effects of Wolbachia on mite reproduction and post-zygotic reproductive isolation are discussed.}, } @article {pmid8799741, year = {1996}, author = {Tsagkarakou, A and Guillemaud, T and Rousset, F and Navajas, M}, title = {Molecular identification of a Wolbachia endosymbiont in a Tetranychus urticae strain (Acari: Tetranychidae).}, journal = {Insect molecular biology}, volume = {5}, number = {3}, pages = {217-221}, doi = {10.1111/j.1365-2583.1996.tb00057.x}, pmid = {8799741}, issn = {0962-1075}, mesh = {Animals ; Bacterial Proteins/*genetics ; Base Sequence ; *Cytoskeletal Proteins ; *DNA, Bacterial ; *DNA, Ribosomal ; Female ; Mites/*microbiology ; Molecular Sequence Data ; Phylogeny ; *RNA, Ribosomal, 16S ; Rickettsiaceae/*genetics ; }, abstract = {Wolbachia, a maternally inherited bacterium, is involved in alterations of arthropod sexuality. Reproductive incompatibilities are often observed in miles, but the existence of this microorganism in their cytoplasm has not yet been demonstrated. We identified the presence of Wolbachia in a strain of the spider mite Tetranychus urticae based on the amplification and sequencing of part of the 16S rDNA and ftsZ genes. In order to establish the phylogenetic relationships between Wolbachia found in T. urticae and in other arthropods, we aligned the resulting sequences with already published ones. For both 16S and ftsZ genes the Wolbachia carried by T. urticae clustered together with Wolbachia found in other arthropods.}, } @article {pmid8764685, year = {1996}, author = {Chang, NW and Wade, MJ}, title = {An improved microinjection protocol for the transfer of Wolbachia pipientis between infected and uninfected strains of the flour beetle Tribolium confusum.}, journal = {Canadian journal of microbiology}, volume = {42}, number = {7}, pages = {711-714}, doi = {10.1139/m96-097}, pmid = {8764685}, issn = {0008-4166}, support = {GM22523/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/microbiology ; Female ; Larva/microbiology/physiology ; Male ; Microinjections ; Ovum/microbiology ; Reproduction ; Rickettsiaceae/*physiology ; Tribolium/*microbiology/physiology ; }, abstract = {The lethality of halocarbon and other oils to hatching larvae of the flour beetle Tribolium confusum limits existing microinjection protocols, because postinjection survivorship is only 5.6% of the eggs injected. We report the development of an oil-free protocol that improves survivorship fivefold. We used this protocol to transfect the cytoplasmic endosymbiont Wolbachia pipientis from infected donor eggs to uninfected host eggs and observed reproductive incompatibility in 40% of the surviving, injected eggs. Compared with mock injected controls (35.9% survival) or microinjection of uninfected cytoplasm (32.3% survival), injection of Wolbachia-infected cytoplasm into host eggs significantly reduced egg-to-adult survival (25.1%).}, } @article {pmid8817452, year = {1996}, author = {Lassy, CW and Karr, TL}, title = {Cytological analysis of fertilization and early embryonic development in incompatible crosses of Drosophila simulans.}, journal = {Mechanisms of development}, volume = {57}, number = {1}, pages = {47-58}, doi = {10.1016/0925-4773(96)00527-8}, pmid = {8817452}, issn = {0925-4773}, mesh = {Animals ; Centrosome/physiology ; Chromosome Aberrations/physiology ; Crosses, Genetic ; Drosophila/*embryology/*microbiology/physiology ; Female ; Fertilization ; Infertility/microbiology ; Male ; Rickettsia ; Sperm Tail/physiology ; }, abstract = {Cytoplasmic incompatibility (CI) is a unique form of male sterility found in numerous insect species that harbor a bacterial endosymbiont Wolbachia. CI is characterized by severe reduction in the progeny produced when infected males are crossed to uninfected females. The reduction in progeny correlates with developmental defects that arise during and immediately following fertilization, suggesting that sperm function is disrupted. To investigate the nature of the cellular defects associated with CI, fertilization and early embryonic development were examined in normal and incompatible crosses of Drosophila simulans using anti-sperm, anti-tubulin and anti-chromatin antibodies. Although pleiotropic, defects associated with CI can be classified into five broad categories: (1) sperm defects in the egg; (2) aberrant morphology of the mitotic apparatus; (3) defects in chromatin structure; (4) proliferation of centrosomes in the absence of nuclear division; and (5) loss of mitotic synchrony. Although mitosis and chromosome behavior are severely disrupted in CI crosses during early development, centrosome duplication and migration appear to continue unabated. The available cytological data suggest that the primary defects observed in incompatible crosses are due to defects in chromosome replication/segregation and in associated centrosome/microtubule-based processes.}, } @article {pmid8725242, year = {1996}, author = {Perrot-Minnot, MJ and Guo, LR and Werren, JH}, title = {Single and double infections with Wolbachia in the parasitic wasp Nasonia vitripennis: effects on compatibility.}, journal = {Genetics}, volume = {143}, number = {2}, pages = {961-972}, pmid = {8725242}, issn = {0016-6731}, mesh = {Animals ; Polymerase Chain Reaction ; Rickettsia/*genetics ; Rickettsia Infections/*genetics ; Wasps/*microbiology ; }, abstract = {Wolbachia are cytoplasmically inherited bacteria responsible for reproductive incompatibility in a wide range of insects. There has been little exploration, however, of within species Wolbachia polymorphisms and their effects on compatibility. Here we show that some strains of the parasitic wasp Nasonia vitripennis are infected with two distinct bacterial strains (A and B) whereas others are singly infected (A or B). Double and single infections are confirmed by both PCR amplification and Southern analysis of genomic DNA. Furthermore, it is shown that prolonged larval diapause (the overwintering stage of the wasp) of a double-infected strain can lead to stochastic loss of one or both bacterial strains. After diapause of a double-infected line, sublines were produced with AB, A only, B only or no Wolbachia. A and B sublines are bidirectionally incompatible, whereas males from AB lines are unidirectionally incompatible with females of A and B sublines. Results therefore show rapid development of bidirectional incompatibility within a species due to segregation of associated symbiotic bacteria.}, } @article {pmid8575931, year = {1996}, author = {Hoffmann, AA and Clancy, D and Duncan, J}, title = {Naturally-occurring Wolbachia infection in Drosophila simulans that does not cause cytoplasmic incompatibility.}, journal = {Heredity}, volume = {76 (Pt 1)}, number = {}, pages = {1-8}, doi = {10.1038/hdy.1996.1}, pmid = {8575931}, issn = {0018-067X}, mesh = {Animals ; Australia ; Base Sequence ; Biological Evolution ; Crosses, Genetic ; Cytoplasm/genetics/*physiology ; DNA Primers ; Drosophila/genetics/*microbiology ; Female ; Fertility/genetics ; Genetics, Population ; Male ; Molecular Sequence Data ; Rickettsiaceae/*physiology ; }, abstract = {Microbes of the genus Wolbachia are transmitted by their hosts via the maternal parent and are responsible for cytoplasmic incompatibility among insect populations. This phenomenon can result in Wolbachia spreading through natural populations as previously demonstrated in Drosophila simulans. Here we describe another Wolbachia infection in D. simulans that does not cause cytoplasmic incompatibility. This is a property of the Wolbachia rather than the nuclear background. The infection occurs at a low frequency in natural populations from eastern Australia. The infection shows perfect maternal transmission in the field and does not cause any detectable deleterious effects on its host. These findings suggest that the Wolbachia infection behaves like a neutral variant in populations. The infection may represent an evolutionary outcome of interactions between Wolbachia infections and their hosts.}, } @article {pmid7489921, year = {1995}, author = {Bensaadi-Merchermek, N and Salvado, JC and Cagnon, C and Karama, S and Mouchès, C}, title = {Characterization of the unlinked 16S rDNA and 23S-5S rRNA operon of Wolbachia pipientis, a prokaryotic parasite of insect gonads.}, journal = {Gene}, volume = {165}, number = {1}, pages = {81-86}, doi = {10.1016/0378-1119(95)00539-i}, pmid = {7489921}, issn = {0378-1119}, mesh = {Animals ; Base Sequence ; Cloning, Molecular ; Culex/microbiology ; DNA, Ribosomal/*genetics ; Molecular Sequence Data ; Operon/*genetics ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; RNA, Ribosomal, 23S/*genetics ; Rickettsiaceae/*genetics ; }, abstract = {The rRNA-encoding genes (rDNAs) have been cloned and characterized from Wolbachia pipientis (Wp), the gonadial bacteria-like parasite of the mosquito Culex pipiens (Cp) and the moth Ephestia cautella (Ec). In Wp from both insect species the rDNAs are organized in a way which appears to be very unusual. The rRNAs are encoded by two unlinked transcription units, each present in a single copy per genome. One contains the 16S rDNA only, while the other is an operon encoding both the 23S and 5S rDNAs. Each transcription unit contains two putative upstream promoters, and downstream a Rho-independent terminator. The 16S rDNA, as well as the 23S-5S rRNA operon are not linked to any tRNA-encoding sequence and lack the antitermination boxes which are usually present immediately downstream from eubacterial promoters of rDNAs. Wp infecting Ec and Cp are highly similar taking as criteria the rDNAs and their flanking sequences. However, it clearly appears that each insect species harbours a different and specific Wp strain, or even subspecies. Phylogenetic relationships deduced from the complete sequences of their rDNAs undoubtedly confirm that Wp from Cp and Ec belong to the alpha-group of Proteobacteria, and are closely related to the Rickettsia.}, } @article {pmid8825761, year = {1995}, author = {Hoshizaki, S and Shimada, T}, title = {PCR-based detection of Wolbachia, cytoplasmic incompatibility microorganisms, infected in natural populations of Laodelphax striatellus (Homoptera: Delphacidae) in central Japan: has the distribution of Wolbachia spread recently?.}, journal = {Insect molecular biology}, volume = {4}, number = {4}, pages = {237-243}, doi = {10.1111/j.1365-2583.1995.tb00029.x}, pmid = {8825761}, issn = {0962-1075}, mesh = {Animals ; Base Sequence ; DNA, Bacterial/*genetics ; DNA, Ribosomal/*genetics ; Insecta/*microbiology ; Japan ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*genetics/isolation & purification ; }, abstract = {Cytoplasmic incompatibility is caused in various insects by intracellular infection with rickettsia-like microorganisms of the genus Wolbachia. In Japan Laodelphax striatellus shows unidirectional cytoplasmic incompatibility between northeastern and southwestern populations. In this study, nine natural populations of L. striatellus collected from central Japan, including the geographic boundary between the two cytotype populations, were analysed for Wolbachia infection by PCR using primers specific to Wolbachia 16S rDNA. The geographic pattern of the infection rates of the southwestern (high) and the northeastern (low or zero) populations broadly resembled that of a previous study of incompatibility. In populations which originated from the boundary regions between the southwestern and northeastern populations, the infected and uninfected cytotypes coexisted. It is suggested that in some populations of L. striatellus, which formerly had been uninfected with Wolbachia, the infection property has changed to the infected. Based on our results, we conclude that the distribution of Wolbachia-infected L. striatellus populations have spread northeasterly during the last 12 years.}, } @article {pmid8719164, year = {1995}, author = {Sironi, M and Bandi, C and Sacchi, L and Di Sacco, B and Damiani, G and Genchi, C}, title = {Molecular evidence for a close relative of the arthropod endosymbiont Wolbachia in a filarial worm.}, journal = {Molecular and biochemical parasitology}, volume = {74}, number = {2}, pages = {223-227}, doi = {10.1016/0166-6851(95)02494-8}, pmid = {8719164}, issn = {0166-6851}, mesh = {Animals ; Arthropods/*microbiology ; Base Sequence ; DNA Primers/genetics ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Dirofilaria/*microbiology/ultrastructure ; Dogs ; Female ; Male ; Microscopy, Electron ; Molecular Sequence Data ; Rickettsiaceae/classification/*genetics/*isolation & purification ; Symbiosis ; }, } @article {pmid8582608, year = {1995}, author = {Merçot, H and Llorente, B and Jacques, M and Atlan, A and Montchamp-Moreau, C}, title = {Variability within the Seychelles cytoplasmic incompatibility system in Drosophila simulans.}, journal = {Genetics}, volume = {141}, number = {3}, pages = {1015-1023}, pmid = {8582608}, issn = {0016-6731}, mesh = {Animals ; Bacteria/classification/genetics ; Base Sequence ; Crosses, Genetic ; Drosophila/classification/*microbiology ; Extrachromosomal Inheritance/genetics ; Female ; Fertility/genetics ; Male ; Molecular Sequence Data ; Polymorphism, Restriction Fragment Length ; Species Specificity ; Symbiosis ; }, abstract = {In Drosophila simulans, we described a cytoplasmic incompatibility (CI) system (Seychelles) restricted to insular populations that harbor the mitochondrial type SiI. Since then, these populations have been shown to be heterogeneous, some being infected by one Wolbachia genetic variant only (wHa), while others are infected simultaneously by wHa and by another variant (wNo) always found in association with wHa. We have experimentally obtained two D. simulans strains only infected by the wNo variant. This variant determines its own cytoplasmic incompatibility type. In particular, the cross between wNo-bearing flies and wHa-bearing ones is bidirectionally incompatible. The Seychelles CI type, stricto sensu, is distinguished by being determined by the simultaneous presence of two Wolbachia variants that we found to be mutually incompatible. In addition, we observed incomplete maternal transmission of the Wolbachia.}, } @article {pmid7498425, year = {1995}, author = {Sinkins, SP and Braig, HR and O'Neill, SL}, title = {Wolbachia pipientis: bacterial density and unidirectional cytoplasmic incompatibility between infected populations of Aedes albopictus.}, journal = {Experimental parasitology}, volume = {81}, number = {3}, pages = {284-291}, doi = {10.1006/expr.1995.1119}, pmid = {7498425}, issn = {0014-4894}, support = {AI28778/AI/NIAID NIH HHS/United States ; AI34355/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/*microbiology ; Animals ; Base Sequence ; Crosses, Genetic ; DNA Primers/chemistry ; Drosophila/microbiology ; Female ; Insect Vectors/genetics/*microbiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Rickettsiaceae/genetics/*growth & development ; }, abstract = {Unidirectional cytoplasmic incompatibility is seen when certain Wolbachia-infected insect populations are crossed. Two hypotheses might explain this phenomenon: superinfections with mutually incompatible strains of Wolbachia producing incompatibility when crossed to individuals infected with only a single bacterial strain or, alternatively, a bacterial dosage model, with differences in Wolbachia densities responsible for the incompatibility. A quantitative PCR assay was set up as a general method to compare Wolbachia densities between populations. Using this assay in unidirectionally incompatible stocks of the mosquito Aedes albopictus, we have determined that densities are significantly higher in Houston than in the Mauritius and Koh Samui stocks. This is consistent with a dosage model for the observed crossing patterns, but does not rule out the possibility that superinfection is the primary cause of the incompatibility.}, } @article {pmid8587875, year = {1995}, author = {Sinkins, SP and Braig, HR and O'Neill, SL}, title = {Wolbachia superinfections and the expression of cytoplasmic incompatibility.}, journal = {Proceedings. Biological sciences}, volume = {261}, number = {1362}, pages = {325-330}, doi = {10.1098/rspb.1995.0154}, pmid = {8587875}, issn = {0962-8452}, support = {AI28778/AI/NIAID NIH HHS/United States ; AI34355/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/microbiology ; Animals ; Base Sequence ; Cell Line ; Crosses, Genetic ; DNA, Bacterial/analysis ; Drosophila/*genetics/*microbiology ; Female ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/genetics/pathogenicity/*physiology ; Superinfection ; Symbiosis ; }, abstract = {Strains of Drosophila simulans from Riverside, California (DSR) and Hawaii (DSH) harbour distinct strains of the cytoplasmic incompatibility microorganism Wolbachia, resulting in the expression of bidirectional incompatibility when crossed. D. simulans lines carrying both of these (superinfected) Wolbachia strains were generated by the transfer of infected DSH cytoplasm into DSR embryos by microinjection. The superinfected flies were unidirectionally incompatible with both DSR and DSH individuals. As a result of this pattern, the superinfected state was observed to replace single infections in laboratory populations. The ability of the superinfection to spread was modulated by the production of singly infected offspring from superinfected mothers: strain segregation was observed under crowded larval rearing conditions. An inverse correlation between the penetrance of the cytoplasmic incompatibility phenotype and the degree of larval crowding was also observed. The findings have implications for the evolution of bidirectionally incompatible strains, and lead to the prediction that superinfections should be relatively common in field populations. Evidence for a natural superinfection in the mosquito Aedes albopictus is discussed. The results also have applied significance for the generation of insect lines capable of driving desirable genes into populations already infected with Wolbachia, thus allowing repeated opportunities for population replacement.}, } @article {pmid7558891, year = {1995}, author = {Werren, JH and Jaenike, J}, title = {Wolbachia and cytoplasmic incompatibility in mycophagous Drosophila and their relatives.}, journal = {Heredity}, volume = {75 (Pt 3)}, number = {}, pages = {320-326}, doi = {10.1038/hdy.1995.140}, pmid = {7558891}, issn = {0018-067X}, mesh = {Animals ; Base Sequence ; Cloning, Molecular ; Crosses, Genetic ; Cytoplasm/*physiology ; DNA Primers ; Drosophila/genetics/*microbiology/physiology ; Female ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Rickettsiaceae/genetics/isolation & purification/*physiology ; }, abstract = {Bacterial symbionts belonging to the genus Wolbachia are associated with postzygotic reproductive incompatibility in a number of insect species. Using polymerase chain reaction (PCR) amplification of bacterial gene sequences, strains from 10 species belonging to the closely related quinaria, testacea and tripunctata groups of Drosophila were screened for the presence of Wolbachia in their reproductive tissues. Those screened included the mycophagous species D. falleni, D. recens, D. phalerata, D. testacea, D. neotestacea, D. orientacea, D. putrida and D. tripunctata, and the nonmycophagous species D. palustris and D. quinaria. Two species, D. recens and D. orientacea, were found to be infected with Wolbachia. Subsequent tests of four additional strains of D. recens found all to be infected with the bacteria. It was established that these bacteria cause partial cytoplasmic incompatibility in D. recens by antibiotic curing followed by crosses between cured and uncured strains. Curing was confirmed by a PCR assay. Although most species of insects shown to be infected with Wolbachia are cosmopolitan and/or have undergone recent range expansion in association with human activity, D. recens and D. orientacea are endemic species with specialized ecological habits. Preliminary molecular phylogenetic analysis indicates that, among the species we examined, D. quinaria is most closely related to D. recens. To determine whether the bacteria are involved in reproductive isolation between these two species, reciprocal crosses were carried out between D. quinaria and both infected and uninfected (cured) strains of D. recens. Although these species did mate with each other, all interspecific crosses failed to yield hybrid progeny, indicating that the bacteria are not responsible for reproductive incompatibility between these species.}, } @article {pmid8589844, year = {1995}, author = {Zchori-Fein, E and Faktor, O and Zeidan, M and Gottlieb, Y and Czosnek, H and Rosen, D}, title = {Parthenogenesis-inducing microorganisms in Aphytis (Hymenoptera: Aphelinidae).}, journal = {Insect molecular biology}, volume = {4}, number = {3}, pages = {173-178}, doi = {10.1111/j.1365-2583.1995.tb00023.x}, pmid = {8589844}, issn = {0962-1075}, mesh = {Animals ; Antibiotics, Antitubercular/pharmacology ; Base Sequence ; DNA Primers ; DNA, Bacterial/analysis ; *DNA, Ribosomal ; Female ; Fertility ; Male ; Molecular Sequence Data ; *Parthenogenesis ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/*physiology ; Rifampin/pharmacology ; Symbiosis ; Wasps/classification/*microbiology/*physiology ; }, abstract = {Production of males in uniparental lines of two species in the parasitic wasp genus Aphytis was induced by rifampicin, and male sexual functioning was determined. Wolbachia-specific 16S rDNA primers were used in a PCR in order to: (1) assess correlation between thelytokous reproduction and the presence of Wolbachia; (2) detect the loss of Wolbachia DNA in uniparental A. lingnanensis following antibiotic treatments, with or without the presence of a host; and (3) clone and sequence part of the Wolbachia 16S rDNA from the uniparental Aphytis species for phylogenetic studies. Males produced viable sperm that was transferred to the female spermatheca following mating. However, sperm failure to effect egg fertilization resulted in all-male progeny. Wolbachia were found in the two uniparental (A. lingnanensis and A. diaspidis) but not in the two biparental (A. lingnanensis and A. melinus) Aphytis lines tested. They can be detected in wasps up to 7 days following antibiotic treatments, regardless of the presence of host. The 16S rDNA for the symbionts in the two Aphytis species is virtually identical, and is most closely related to the Wolbachia found in Muscidifurax uniraptor (Pteromalidae).}, } @article {pmid7498773, year = {1995}, author = {Turelli, M and Hoffmann, AA}, title = {Cytoplasmic incompatibility in Drosophila simulans: dynamics and parameter estimates from natural populations.}, journal = {Genetics}, volume = {140}, number = {4}, pages = {1319-1338}, pmid = {7498773}, issn = {0016-6731}, mesh = {Africa, Southern ; Age Factors ; Americas ; Animals ; Base Sequence ; California ; Crosses, Genetic ; Cytoplasm ; DNA, Mitochondrial/genetics ; Drosophila/genetics/*microbiology/physiology ; Female ; Florida ; Genetics, Population ; Hawaii ; Infertility/microbiology ; Male ; Molecular Sequence Data ; New Caledonia ; Polymerase Chain Reaction ; Rickettsiaceae/*physiology ; Spermatozoa/microbiology ; }, abstract = {In Drosophila simulans, cytoplasmically transmitted Wolbachia microbes cause reduced egg hatch when infected males mate with uninfected females. A Wolbachia infection and an associated mtDNA variant have spread northward through California since 1986. PCR assays show that Wolbachia infection is prevalent throughout the continental US and Central and South America, but some lines from Florida and Ecuador that are PCR-positive for Wolbachia do not cause incompatibility. We estimate from natural populations infection frequencies and the transmission and incompatibility parameter values that affect the spread of the infection. On average, infected females from nature produce 3-4% uninfected ova. Infected females with relatively low fidelity of maternal transmission show partial incompatibility with very young infected laboratory males. Nevertheless, crosses between infected flies in nature produce egg-hatch rates indistinguishable from those produced by crosses between uninfected individuals. Incompatible crosses in nature produce hatch rates 30-70% as high as those from compatible crosses. Wild-caught infected and uninfected females are equally fecund in the laboratory. Incompatibility decreases with male age, and age-specific incompatibility levels suggest that males mating in nature may often be 2 or 3 weeks old. Our parameter estimates accurately predict the frequency of Wolbachia infection in California populations.}, } @article {pmid7498772, year = {1995}, author = {Giordano, R and O'Neill, SL and Robertson, HM}, title = {Wolbachia infections and the expression of cytoplasmic incompatibility in Drosophila sechellia and D. mauritiana.}, journal = {Genetics}, volume = {140}, number = {4}, pages = {1307-1317}, pmid = {7498772}, issn = {0016-6731}, support = {AI-34355/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Arthropods/classification/microbiology ; Crosses, Genetic ; Cytoplasm ; Drosophila/classification/genetics/*microbiology ; Female ; Infertility/microbiology ; Male ; Molecular Sequence Data ; Phylogeny ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/classification/genetics/*physiology ; Species Specificity ; Spermatozoa/microbiology ; }, abstract = {Various stocks of Drosophila mauritiana and D. sechellia were found to be infected with Wolbachia, a Rickettsia-like bacterium that is known to cause cytoplasmic incompatibility and other reproductive abnormalities in arthropods. Testing for the expression of cytoplasmic incompatibility in these two species showed partial incompatibility in D. sechellia but no expression of incompatibility in D. mauritiana. To determine whether absence of cytoplasmic incompatibility in D. mauritiana was due to either the bacterial or host genome, we transferred bacteria from D. mauritiana into an uninfected strain of D. simulans, a host species known to express high levels of incompatibility with endogenous Wolbachia. We also performed the reciprocal transfer of the natural D. simulans Riverside infection into a tetracycline-treated stock of D. mauritiana. In each case, the ability to express incompatibility was unaltered by the different host genetic background. These experiments indicate that in D. simulans and D. mauritiana expression of the cytoplasmic incompatibility phenotype is determined by the bacterial strain and that D. mauritiana harbors a neutral strain of Wolbachia.}, } @article {pmid7644549, year = {1995}, author = {Werren, JH and Zhang, W and Guo, LR}, title = {Evolution and phylogeny of Wolbachia: reproductive parasites of arthropods.}, journal = {Proceedings. Biological sciences}, volume = {261}, number = {1360}, pages = {55-63}, doi = {10.1098/rspb.1995.0117}, pmid = {7644549}, issn = {0962-8452}, mesh = {Animals ; Arthropods/*microbiology ; Base Sequence ; Cloning, Molecular ; DNA Primers ; Molecular Sequence Data ; *Phylogeny ; Polymerase Chain Reaction ; Rickettsiaceae/classification/*genetics/pathogenicity ; }, abstract = {Wolbachia are cytoplasmically inherited bacteria found in reproductive tissues of many arthropod species. These bacteria are associated with reproductive alterations in their hosts, including parthenogenesis, reproductive incompatibility and feminization. A fine-scale phylogenetic analysis was done using DNA sequences from ftsZ, a rapidly evolving bacterial cell-cycle gene. ftsZ sequences were determined for 38 different Wolbachia strains from 31 different species of insects and one isopod. The following results were found: (i) there are two major division of Wolbachia (A and B) which diverged 58-67 millions years before present based upon synonymous substitution rates; (ii) a general concordance is found between the ftsZ and 16S rDNA phylogenies, indicating that these represent bacterial strain (rather than simply gene) phylogenies; however, a possible example of recombination between A and B division bacteria may have occurred in the feminizing Wolbachia present in an isopod; (iii) extensive horizontal transmission of Wolbachia has occurred between insect taxa, including different insect orders; one strain in particular (designated Adm) shows extensive recent horizontal transmission; (iv) there is an association between the Wolbachia found in a parasitic wasp (Nasonia) and its fly host (Protocalliphora), suggesting exchange of bacteria between these species; (v) parthenogenesis induction has evolved several times among the Wolbachia; and (vi) some insects harbour infections with more than one Wolbachia strain, even within individual insects.}, } @article {pmid7604001, year = {1995}, author = {Rousset, F and Solignac, M}, title = {Evolution of single and double Wolbachia symbioses during speciation in the Drosophila simulans complex.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {92}, number = {14}, pages = {6389-6393}, pmid = {7604001}, issn = {0027-8424}, mesh = {Animals ; Base Sequence ; *Biological Evolution ; DNA, Bacterial/genetics ; DNA, Mitochondrial/*genetics ; DNA, Ribosomal/genetics ; Drosophila/*genetics/*microbiology ; Female ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Restriction Mapping ; Rickettsiaceae/genetics/*physiology ; Species Specificity ; *Symbiosis ; }, abstract = {Maternally inherited bacteria of the genus Wolbachia are responsible for the early death of embryos in crosses between uninfected females and infected males in several insect species. This phenomenon, known as cytoplasmic incompatibility, also occurs between strains infected by different symbionts in some species, including Drosophila simulans. Wolbachia was found in two species closely related to D. simulans, Drosophila mauritiana, and Drosophila sechellia, and shown to cause incompatibility in the latter species but not in D. mauritiana. Comparison of bacterial and mtDNA history clarifies the origins of bacterial and incompatibility polymorphisms in D. simulans. Infection in D. mauritiana is probably the result of introgression of an infected D. simulans cytoplasm. Some D. simulans and D. sechellia cytoplasmic lineages harbor two bacteria as a consequence of a double infection which probably occurred in a common ancestor. The descendant symbionts in each species are associated with similar incompatibility relationships, which suggests that little variation of incompatibility types has occurred within maternal lineages beyond that related to the density of symbionts in their hosts.}, } @article {pmid8525055, year = {1995}, author = {Roux, V and Raoult, D}, title = {Phylogenetic analysis of the genus Rickettsia by 16S rDNA sequencing.}, journal = {Research in microbiology}, volume = {146}, number = {5}, pages = {385-396}, doi = {10.1016/0923-2508(96)80284-1}, pmid = {8525055}, issn = {0923-2508}, mesh = {Base Sequence ; In Vitro Techniques ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/*chemistry/genetics ; Rickettsia/*classification/genetics ; }, abstract = {Rickettsiae are Gram-negative bacteria which multiply only inside host cells and need arthropods either as reservoirs or as vectors. Using the polymerase chain reaction and an automated laser fluorescent DNA sequencer, we amplified and sequenced the 16S rRNA (rDNA) of all available bacteria of the genus Rickettsia. R. tsutsugamushi remained close to the other bacteria of the genus Rickettsia using this technique, contrary to previous conclusions based on the study of the Sta-58 protein antigen. We found that R. canada was not included in the typhus group, as is currently recognized, but was grouped with the rickettsiae of the spotted fever group (SFG). All the SFG rickettsiae tested were grouped in the same cluster (R. conorii, Indian tick typhus rickettsia, Astrakhan fever rickettsia, Israeli tick typhus rickettsia, HA-91, R. sibirica, R. parkeri, "R. africae", "R. slovaca", R. rickettsii, Thai tick typhus rickettsia, R. japonica, R. massiliae, R. rhipicephali, R. montana, two recent isolates GS and Bar 29, R. australis, R. akari, R. bellii and R. helvetica). The recently described ELB bacterium, the agent of the Californian murine typhus, and AB bacterium, a bacterium associated with male killing in the ladybird beetle, were found in this cluster. The sequences of R. conorii Moroccan strain/Indian tick typhus rickettsia, R. massiliae/GS and R. sibirica/HA-91 were identical. All the rickettsiae had a unique ancestor with bacteria also isolated in arthropods (Ehrlichia, Cowdria, Anaplasma, Wolbachia pipientis), eventually pathogenic for mammals and implicated in parthenogenesis and cytoplasmic incompatibility. We conclude that a unique bacterium started a stable association with arthropod ancestors and generated the observed diversity of the currently isolated members of the Rickettsiales.}, } @article {pmid7547474, year = {1995}, author = {Kose, H and Karr, TL}, title = {Organization of Wolbachia pipientis in the Drosophila fertilized egg and embryo revealed by an anti-Wolbachia monoclonal antibody.}, journal = {Mechanisms of development}, volume = {51}, number = {2-3}, pages = {275-288}, doi = {10.1016/0925-4773(95)00372-x}, pmid = {7547474}, issn = {0925-4773}, support = {R01 GM47962/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Antibodies, Monoclonal ; Blastoderm/microbiology ; Centrosome/microbiology ; Colony Count, Microbial ; Drosophila/embryology/*microbiology ; Female ; Fluorescent Antibody Technique, Indirect ; Male ; Microtubules/microbiology ; Mitosis ; Ovum/microbiology ; Rickettsiaceae/*immunology/*isolation & purification ; Symbiosis ; }, abstract = {Cytoplasmic incompatibility (CI) in Drosophila is related to the presence of Wolbachia, an intracellular microorganism found in many species of insects. In order to study the intracellular localization of Wolbachia in eggs and embryos, we have purified the bacteria from fly embryos and subsequently generated a monoclonal antibody (Mab Wol-1) specific for Wolbachia. Indirect immunofluorescence staining using Wol-1 reveals that during mitosis, Wolbachia are localized near spindle poles and centrosomes. Double label immunofluorescence experiments using anti-tubulin and anti-Wolbachia antibodies show that Wolbachia co-localize with centrosomal microtubules throughout the cell cycle. Direct interactions between the bacteria and centrosome-organized microtubules are implied from seven observations: (1) throughout the mitotic cycle, the position and movement of Wolbachia precisely mimic the behavior of the centrosome and apparently associated with centrosome-organized microtubules; (2) Wolbachia segregate equally to each spindle pole during mitosis; (3) Wolbachia do not associate with spindle microtubules during mitosis; (4) Wolbachia located in the egg cortex localize to the domains of cytoplasm organized by microtubules during blastoderm formation; (5) polar body nuclei that lack centrosomes but contain associated microtubules do not contain Wolbachia; (6) Wolbachia no longer associated with yolk nuclei, following differentiation and loss of centrosomes; (7) during pole cell formation, Wolbachia co-localize with the centrosome on the apical side of the nucleus as pole cells form. Quantitative data indicates that no Wolbachia growth occurs during the preblastoderm period even though rapid nuclear, and subsequent cellular, proliferation takes place during this same period. This indicates that Wolbachia are under strict growth regulation by the host suggesting that host factors play a role in regulating growth of Wolbachia in the egg. Further cellular and molecular studies of the extensive, global interactions between host and symbiont observed in this egg should provide important new insights into the evolution of host/symbiosis and the cell biology of cytoplasmic incompatibility.}, } @article {pmid7598906, year = {1995}, author = {Reed, KM and Werren, JH}, title = {Induction of paternal genome loss by the paternal-sex-ratio chromosome and cytoplasmic incompatibility bacteria (Wolbachia): a comparative study of early embryonic events.}, journal = {Molecular reproduction and development}, volume = {40}, number = {4}, pages = {408-418}, doi = {10.1002/mrd.1080400404}, pmid = {7598906}, issn = {1040-452X}, support = {R01-HD24310/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; Bacteria/*genetics ; Embryo, Nonmammalian/microbiology ; Gene Expression Regulation, Bacterial ; Gene Expression Regulation, Developmental ; Genome ; Male ; Sex Chromosomes/*genetics ; Wasps/*embryology/genetics/microbiology ; }, abstract = {Paternal genome loss (PGL) during early embryogenesis is caused by two different genetic elements in the parasitoid wasp, Nasonia vitripennis. Paternal sex ratio (PSR) is a paternally inherited supernumerary chromosome that disrupts condensation of the paternal chromosomes by the first mitotic division of fertilized eggs. Bacteria belonging to the genus Wolbachia are present in Nasonia eggs and also disrupt paternal chromosome condensation in crosses between cytoplasmically incompatible strains. Cytoplasmic incompatibility Wolbachia are widespread in insects, whereas PSR is specific to this wasp. PGL results in production of male progeny in Nasonia due to haplodiploid sex determination. The cytological events associated with PGL induced by the PSR chromosome and by Wolbachia were compared by fluorescent light microscopy using the fluorochrome Hoescht 33258. Cytological examination of eggs fertilized with PSR-bearing sperm revealed that a dense paternal chromatin mass forms prior to the first metaphase. Quantification of chromatin by epifluorescence indicates that this mass does undergo replication along with the maternal chromatin prior to the first mitotic division but does not replicate during later mitotic cycles. Contrary to previous reports using other staining methods, the paternal chromatin mass remains condensed during interphase and persists over subsequent mitotic cycles, at least until formation of the syncytial blastoderm and cellularization, at which time it remains near the center of the egg with the yolk nuclei. Wolbachia-induced PGL shows several marked differences. Most notable is that the paternal chromatin mass is more diffuse and tends to be fragmented during the first mitotic division, with portions becoming associated with the daughter nuclei. Nuclei containing portions of the paternal chromatin mass appear to be delayed in subsequent mitotic divisions relative to nuclei free of paternal chromatin. Crosses combining incompatibility with PSR were cytologically similar to Wolbachia-induced PGL, although shearing of the paternal chromatin mass was reduced. Wolbachia may, therefore, block an earlier stage of paternal chromatin processing in the fertilized eggs than does PSR.}, } @article {pmid7800041, year = {1995}, author = {Wade, MJ and Chang, NW}, title = {Increased male fertility in Tribolium confusum beetles after infection with the intracellular parasite Wolbachia.}, journal = {Nature}, volume = {373}, number = {6509}, pages = {72-74}, doi = {10.1038/373072a0}, pmid = {7800041}, issn = {0028-0836}, mesh = {Animals ; Female ; *Fertility ; Male ; Rickettsiaceae/*physiology ; Symbiosis ; Tribolium/genetics/microbiology/*physiology ; }, abstract = {The cytoplasmically inherited microorganism Wolbachia pipientis behaves like a sexually selected trait in its host, the flour beetle Tribolium confusum, enhancing male fertility at the expense of female fecundity. Here we show that infected females have fewer offspring than uninfected females but infected males have a large fertility advantage over uninfected males within multiply-inseminated infected or uninfected females. The male fertility effect accelerates the spread of the Wolbachia through the host population and expands the initial opportunity for hitch-hiking of host nuclear genes. Sperm competition in a host, mediated by endosymbionts, has not been previously described.}, } @article {pmid7894745, year = {1994}, author = {Bourtzis, K and Nirgianaki, A and Onyango, P and Savakis, C}, title = {A prokaryotic dnaA sequence in Drosophila melanogaster: Wolbachia infection and cytoplasmic incompatibility among laboratory strains.}, journal = {Insect molecular biology}, volume = {3}, number = {3}, pages = {131-142}, doi = {10.1111/j.1365-2583.1994.tb00160.x}, pmid = {7894745}, issn = {0962-1075}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/*genetics ; Base Sequence ; Crosses, Genetic ; DNA-Binding Proteins/*genetics ; Drosophila/classification/genetics/microbiology ; Drosophila melanogaster/genetics/*microbiology ; *Extrachromosomal Inheritance ; Female ; *Genes, Bacterial ; Infertility/microbiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Alignment ; Tetracycline/therapeutic use ; }, abstract = {Using oligonucleotide primers derived from the aligned polypeptide sequences of several prokaryotic dnaA genes, we amplified from Drosophila melanogaster DNA a 557 bp fragment containing a single open reading frame. The predicted peptide sequence shows a significant similarity to previously characterized protein sequences that are encoded by the dnaA genes of several prokaryotes. The dnaA sequences are also detectable by PCR in DNA from Drosophila simulans and Nasonia vitripennis flies which are infected by a symbiotic bacterium assigned to the type species Wolbachia pipientis. A tetracycline treatment that eradicates bacterial parasites from insects, abolishes the dnaA sequences from Drosophila and Nasonia DNA. In addition, dnaA-positive Drosophila melanogaster contain numerous rod-shaped bacteria in embryos, which are abolished in subsequent generations after treatment with tetracycline. Combined with phylogenetic analysis of DnaA and 16S rRNA sequences, these results show that the dnaA cognate comes from Wolbachia. A survey of Drosophila stocks using PCR amplification of dnaA and 16S rRNA sequences showed that Wolbachia is widely spread among D. melanogaster laboratory strains but absent from several established strains of the Mediterranean fruit fly Ceratitis capitata. Evidence is also presented that presence of the bacterium can cause partial cytoplasmic incompatibility between infected and non-infected D. melanogaster strains.}, } @article {pmid8200813, year = {1994}, author = {Rousset, F and de Stordeur, E}, title = {Properties of Drosophila simulans strains experimentally infected by different clones of the bacterium Wolbachia.}, journal = {Heredity}, volume = {72 (Pt 4)}, number = {}, pages = {325-331}, doi = {10.1038/hdy.1994.48}, pmid = {8200813}, issn = {0018-067X}, mesh = {Animals ; Base Sequence ; Crosses, Genetic ; DNA Primers ; Drosophila/microbiology/*physiology ; Female ; Male ; Molecular Sequence Data ; Reproduction/physiology ; Rickettsiaceae/genetics/isolation & purification/*physiology ; }, abstract = {Maternally inherited bacteria of the genus Wolbachia are responsible for reproductive incompatibilities between strains of Drosophila simulans. Such incompatibilities are known in several types of crosses, including both directions of crossing between two types of infected strains, 'R' and 'S'. To determine whether the bidirectional incompatibility between R and S flies is due only to differences between their bacteria, flies from an uninfected strain have been experimentally infected with bacteria associated with each type. The incompatibility properties of experimental strains are close to those of original strains harbouring the same bacteria and therefore independent of nuclear background. Backcross experiments, however, show that the infection level of a strain depends on the nature of paternal ancestors. This is not explained by nuclear effects but is possibly the result of an interaction between the infection levels of both parents, in which the infection level of S strains is an equilibrium between a tendency for females to produce weakly infected offspring and selection of more infected eggs by sperm from infected males.}, } @article {pmid7909859, year = {1994}, author = {Wade, MJ and Stevens, L}, title = {The effect of population subdivision on the rate of spread of parasite-mediated cytoplasmic incompatibility.}, journal = {Journal of theoretical biology}, volume = {167}, number = {1}, pages = {81-87}, doi = {10.1006/jtbi.1994.1052}, pmid = {7909859}, issn = {0022-5193}, support = {GM22523/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*parasitology ; Bacterial Infections/transmission ; Cytoplasm/physiology ; Female ; *Host-Parasite Interactions ; Male ; Models, Biological ; Polymorphism, Restriction Fragment Length ; Population ; Reproduction/*physiology ; }, abstract = {Over 20 species of arthropods are known to harbor an endosymbiotic bacterium, Wolbachia associated with partial reproductive incompatibility. The micro-organism spreads through a random mating population with positive frequency dependence. Many insect populations, however, are subdivided to some degree. In this paper we model the effect of population subdivision on the rate of spread of Wolbachia and show that it slows the rate of spread of the micro-organism but by itself cannot cause a stable polymorphism. The effects of the micro-organism on host fitness, partial incompatibility, and environmental curing are also investigated, both for panmictic and subdivided populations. In all cases, population subdivision slows the rate of spread relative to a panmictic population, but does not produce a stable polymorphism of infection.}, } @article {pmid8005448, year = {1994}, author = {Hoffmann, AA and Clancy, DJ and Merton, E}, title = {Cytoplasmic incompatibility in Australian populations of Drosophila melanogaster.}, journal = {Genetics}, volume = {136}, number = {3}, pages = {993-999}, pmid = {8005448}, issn = {0016-6731}, mesh = {Alphaproteobacteria/isolation & purification/pathogenicity ; Animals ; Australia ; Crosses, Genetic ; Cytoplasm/microbiology ; Drosophila melanogaster/*genetics/microbiology ; Female ; Fluorescent Dyes ; Genetics, Population ; Indoles ; Infertility/genetics/microbiology ; Male ; Polymorphism, Genetic ; }, abstract = {In Drosophila melanogaster, weak incompatibility in crosses between infected and uninfected strains is associated with a Wolbachia microorganism. Crosses between infected males and uninfected females show a reduction (15-30%) in egg hatch. Progeny tests indicated that the infection is widespread in Australian D. melanogaster populations and that populations are polymorphic for the presence of the infection. The infection status of 266 lines from 12 populations along the eastern coast of Australia was determined by 4',6-diamidino-2-phenylindole (DAPI) staining of embryos. All populations contained both infected and uninfected flies. Infection frequencies varied between populations but there was no discernible geographical pattern. Laboratory experiments indicated that the infection was not associated with a reduction in fecundity as in Drosophila simulans. Incompatibility levels could not be increased by laboratory selection on isofemale lines. Factors contributing to the persistence of the infection in D. melanogaster populations are discussed.}, } @article {pmid7906391, year = {1994}, author = {Braig, HR and Guzman, H and Tesh, RB and O'Neill, SL}, title = {Replacement of the natural Wolbachia symbiont of Drosophila simulans with a mosquito counterpart.}, journal = {Nature}, volume = {367}, number = {6462}, pages = {453-455}, doi = {10.1038/367453a0}, pmid = {7906391}, issn = {0028-0836}, mesh = {Aedes/*microbiology ; Animals ; Crosses, Genetic ; DNA, Mitochondrial/analysis ; Drosophila/*microbiology ; Female ; Male ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S/genetics ; Reproduction/physiology ; Rickettsia/genetics/*physiology ; *Symbiosis ; }, abstract = {Inherited rickettsial symbionts of the genus Wolbachia occur commonly in arthropods and have been implicated in the expression of parthenogenesis, feminization and cytoplasmic incompatibility Wolbachia from the Asian tiger mosquito, Aedes albopictus, to replace the natural infection of Drosophila simulans by means of embryonic microinjection techniques. The transferred Wolbachia infection behaves like a natural Drosophila infection with regard to its inheritance, cytoskeleton interactions and ability to induce incompatibility when crossed with uninfected flies. The transinfected flies are bidirectionally incompatible with all other naturally infected strains of Drosophila simulans, however, and as such represent a unique crossing type. The successful transfer of this symbiont between distantly related hosts suggests that it may be possible to introduce this agent experimentally into arthropod species of medical and agricultural importance in order to manipulate natural populations genetically.}, } @article {pmid8123561, year = {1994}, author = {Forsman, M and Sandström, G and Sjöstedt, A}, title = {Analysis of 16S ribosomal DNA sequences of Francisella strains and utilization for determination of the phylogeny of the genus and for identification of strains by PCR.}, journal = {International journal of systematic bacteriology}, volume = {44}, number = {1}, pages = {38-46}, doi = {10.1099/00207713-44-1-38}, pmid = {8123561}, issn = {0020-7713}, mesh = {Base Sequence ; DNA, Bacterial/*chemistry ; DNA, Ribosomal/*chemistry ; Francisella/classification/*genetics/isolation & purification ; Molecular Sequence Data ; Phylogeny ; *Polymerase Chain Reaction ; RNA, Ribosomal, 16S/*genetics ; }, abstract = {The 16S ribosomal DNAs (rDNAs) of two strains of Francisella tularensis and one strain of Francisella philomiragia were sequenced. On the basis of phylogenetic analysis data, the genus Francisella was placed in the gamma subclass of the Proteobacteria. The most closely related organism was the intracellular bacterium Wolbachia persica. The sequenced 16S rDNA molecules of the Francisella species exhibited very high levels of similarity (98.5 to 99.9%). Two variable regions, comprising 390 to 450 nucleotides of the 16S rDNA molecules of 17 additional Francisella strains, including members of the species F. tularensis and F. philomiragia, were also sequenced. At most, six nucleotide differences were observed among the sequences of the F. tularensis strains. The sequence of Francisella novicida was virtually identical to the sequences of the F. tularensis strains, thereby supporting the hypothesis that these organisms are members of the same species. On the basis of the observed differences, primer pairs were designed to distinguish strains by using the PCR at the genus, species, and subspecies levels. This permitted sensitive identification of strains belonging to the genus Francisella and discrimination of the species F. tularensis and F. philomiragia.}, } @article {pmid8117465, year = {1994}, author = {Drancourt, M and Raoult, D}, title = {Taxonomic position of the rickettsiae: current knowledge.}, journal = {FEMS microbiology reviews}, volume = {13}, number = {1}, pages = {13-24}, doi = {10.1111/j.1574-6976.1994.tb00032.x}, pmid = {8117465}, issn = {0168-6445}, mesh = {Alphaproteobacteria/*classification/genetics ; Phenotype ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*classification/genetics ; }, abstract = {The term rickettsiae initially encompassed all intracellular bacteria. Early rickettsial taxonomy was based on a comparison of a few phenotypic characteristics and recently, molecular studies brought new bases for rickettsial taxonomy. All rickettsial species studied so far belong to the alpha and gamma groups of the Proteobacteria. Ehrlichiae complex groups Cowdria ruminantium, Anaplasma marginale and Wolbachia pipientis and the related parthenogenesis and cytoplasmic incompatibility bacteria, whereas Rochalimaea species group with Bartonella bacilliformis. Rickettsia tsutsugamushi may form an independent lineage, whereas molecular data allow to regroup serologically defined typhus and spotted fever group rickettsiae. The true scale of Rickettsia and Coxiella genera remain to be determined.}, } @article {pmid7994122, year = {1994}, author = {Kane, MD and Pierce, NE}, title = {Diversity within diversity: molecular approaches to studying microbial interactions with insects.}, journal = {EXS}, volume = {69}, number = {}, pages = {509-524}, doi = {10.1007/978-3-0348-7527-1_29}, pmid = {7994122}, issn = {1023-294X}, mesh = {Animals ; Bacteria/genetics ; Biological Evolution ; Ecology ; Insecta/genetics/*microbiology ; Parthenogenesis/genetics ; Phylogeny ; Rickettsiaceae/genetics ; Symbiosis/*genetics ; }, abstract = {DNA sequence information has greatly augmented the number of characters available for analysis in phylogenetic research. Nowhere is this more evident than in studies of microbial evolution. Ribosomal DNA sequence data has simultaneously permitted the distinction between individual species and the inference of their phylogenetic relationships in many cases where both were formerly impossible. These have contributed to our understanding of the ecology of particular microbe-host interactions and the history of these relationships over evolutionary time. We describe examples from two ends of the ecological spectrum in insect/bacterial associations: one in which bacteria mediate host cytoplasmic incompatibility and parthenogenesis, and the other in which mycetocyte bacteria augment host nutrition. In the former, the pattern of bacterial interaction is general, with the same or closely related strains of the genus Wolbachia associating with a wide range of insect taxa. In the latter, concordance between host and microbe phylogenies suggests cospeciation between bacteria and host, although it is as yet unclear whether this process has involved step-wise, reciprocal coevolution. We conclude with a discussion of how developments in molecular techniques may aid in analyzing more complex interactions between insects and microbes.}, } @article {pmid8268495, year = {1993}, author = {O'Neill, SL and Gooding, RH and Aksoy, S}, title = {Phylogenetically distant symbiotic microorganisms reside in Glossina midgut and ovary tissues.}, journal = {Medical and veterinary entomology}, volume = {7}, number = {4}, pages = {377-383}, doi = {10.1111/j.1365-2915.1993.tb00709.x}, pmid = {8268495}, issn = {0269-283X}, support = {AI-28778/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Base Sequence ; DNA Primers/chemistry ; DNA, Bacterial/analysis/chemistry ; Female ; Intestines/microbiology ; Molecular Sequence Data ; Ovary/microbiology ; Phylogeny ; Polymerase Chain Reaction ; Rickettsiaceae/genetics/*isolation & purification ; Symbiosis ; Tsetse Flies/*microbiology ; }, abstract = {Many blood-feeding insects, including tsetse flies (Diptera: Glossinidae), harbour intracellular bacterial symbionts. Using isolates from tissues of several Glossina species and diagnostic DNA oligonucleotide primers, a polymerase chain reaction (PCR) based assay was designed to identify symbiotic bacteria. Those inhabiting the midgut of Glossina were found to belong to the gamma subdivision, whereas ovarian Proteobacteria were of the alpha subdivision--probably genus Wolbachia (Rickettsiaceae). The presence of Wolbachia-like Rickettsia in the ovaries of G.morsitans subspecies may help to explain the maternally inherited incompatibility of some crosses within this species.}, } @article {pmid7691685, year = {1993}, author = {Holden, PR and Jones, P and Brookfield, JF}, title = {Evidence for a Wolbachia symbiont in Drosophila melanogaster.}, journal = {Genetical research}, volume = {62}, number = {1}, pages = {23-29}, doi = {10.1017/s0016672300031529}, pmid = {7691685}, mesh = {Animals ; Base Sequence ; Drosophila melanogaster/*microbiology ; Female ; Genes, Bacterial/*genetics ; Male ; Molecular Sequence Data ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/genetics/*isolation & purification ; }, abstract = {The bacterial cell division gene, ftsZ, was used as a specific probe to show the presence of a symbiotic bacterium in two wild type strains of Drosophila melanogaster. Under stringent hybridization conditions we have shown that the bacterium is transferred to the progeny of these strains from infected mothers and can be eradicated by treatment with the antibiotic tetracycline. We have characterized this bacterium, by amplifying and sequencing its 16S rRNA gene, as being a member of the genus Wolbachia, an organism that is known to parasitize a range of insects including Drosophila simulans. In a series of reciprocal crosses no evidence was found that the symbiont causes cytoplasmic incompatibility (CI) which is known to occur in infected strains of D. simulans. The implications of these findings are discussed.}, } @article {pmid7689140, year = {1993}, author = {Holden, PR and Brookfield, JF and Jones, P}, title = {Cloning and characterization of an ftsZ homologue from a bacterial symbiont of Drosophila melanogaster.}, journal = {Molecular & general genetics : MGG}, volume = {240}, number = {2}, pages = {213-220}, pmid = {7689140}, issn = {0026-8925}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; Blotting, Southern ; Cloning, Molecular ; *Cytoskeletal Proteins ; DNA, Bacterial ; Drosophila melanogaster/*microbiology ; GTP-Binding Proteins/*genetics/metabolism ; Larva ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsieae/*genetics/metabolism/physiology ; Sequence Homology, Amino Acid ; Symbiosis ; Tetracycline/pharmacology ; }, abstract = {A 1194 bp open reading frame that codes for a 398 amino acid peptide was cloned from a lambda gt11 library of Drosophila melanogaster genomic DNA. The predicted peptide sequence is very similar to three previously characterized protein sequences that are encoded by the ftsZ genes in Escherichia coli, Bacillus subtilis and Rhizobium meliloti. The FtsZ protein has a major role in the initiation of cell division in prokaryotic cells. Using a tetracycline treatment that eradicates bacterial parasites from insects, the ftsZ homologue has been found to be derived from a bacterium that lives within the D. melanogaster strain. However, polymerase chain reaction (PCR) amplification of the gene from treated embryos suggests that it is not derived from a gut bacterium. Nevertheless, by amplifying and characterizing part of the 16S rRNA from this bacterium we have been able to demonstrate that it is a member of the genus Wolbachia, a parasitic organism that infects, and disturbs the sexual cycle of various strains of Drosophila simulans. We suggest that this ftsZ homologue is implicated in the cell division of Wolbachia, an organism that fails to grow outside the host organism. Sequence and alignment analysis of this ftsZ homologue show the presence of a potential GTP-binding motif indicating that it may function as a GTPase. The consequences of this function particularly with respect to its role in cell division are discussed.}, } @article {pmid8511587, year = {1993}, author = {Boyle, L and O'Neill, SL and Robertson, HM and Karr, TL}, title = {Interspecific and intraspecific horizontal transfer of Wolbachia in Drosophila.}, journal = {Science (New York, N.Y.)}, volume = {260}, number = {5115}, pages = {1796-1799}, doi = {10.1126/science.8511587}, pmid = {8511587}, issn = {0036-8075}, mesh = {Animals ; Cytoplasm/microbiology/physiology ; Drosophila/*microbiology/physiology ; Drosophila melanogaster/*microbiology/physiology ; Female ; Male ; Microinjections ; Microscopy ; Ovum/microbiology/physiology ; Rickettsiaceae/*physiology ; }, abstract = {Cytoplasmic incompatibility (CI) in Drosophila simulans is related to infection of the germ line by a rickettsial endosymbiont (genus Wolbachia). Wolbachia were transferred by microinjection of egg cytoplasm into uninfected eggs of both D. simulans and D. melanogaster to generate infected populations. Transinfected strains of D. melanogaster with lower densities of Wolbachia than the naturally infected D. simulans strain did not express high levels of CI. However, transinfected D. melanogaster egg cytoplasm, transferred back into D. simulans, generated infected populations that expressed CI at levels near those of the naturally infected strain. A transinfected D. melanogaster line selected for increased levels of CI expression also displayed increased symbiont densities. These data suggest that a threshold level of infection is required for normal expression of CI and that host factors help determine the density of the symbiont in the host.}, } @article {pmid8510117, year = {1993}, author = {Yano, Y and Takada, N and Fujita, H}, title = {Ultrastructure of spotted fever rickettsialike microorganisms observed in tissues of Dermacentor taiwanensis (Acari: Ixodidae).}, journal = {Journal of medical entomology}, volume = {30}, number = {3}, pages = {579-585}, doi = {10.1093/jmedent/30.3.579}, pmid = {8510117}, issn = {0022-2585}, mesh = {Animals ; Dermacentor/*microbiology ; Female ; Intestines/microbiology ; Malpighian Tubules/microbiology ; Microscopy, Electron ; Ovary/microbiology ; Rickettsia/isolation & purification/*ultrastructure ; Salivary Glands/microbiology ; }, abstract = {Tissues of midguts, Malpighian tubules, salivary glands, and ovaries of female Dermacentor taiwanensis Sugimoto collected in an area endemic for Japanese spotted fever were examined with an electron microscope, and the presence of two kinds of rickettsialike microorganisms was confirmed in Japan. Based on ultrastructural findings, a rod-shaped organism detected in all the tissues is a Rickettsia; another, irregularly shaped organism that was found only in the ovary, is a Wolbachia. A hemolymph test using a specific immunostain indicated that the Rickettsia belongs to the spotted fever group of rickettsiae.}, } @article {pmid7689622, year = {1993}, author = {Bressac, C and Rousset, F}, title = {The reproductive incompatibility system in Drosophila simulans: DAPI-staining analysis of the Wolbachia symbionts in sperm cysts.}, journal = {Journal of invertebrate pathology}, volume = {61}, number = {3}, pages = {226-230}, doi = {10.1006/jipa.1993.1044}, pmid = {7689622}, issn = {0022-2011}, mesh = {Aging ; Animals ; Drosophila/*microbiology ; Fluorescent Dyes ; Indoles ; Male ; Rickettsiaceae/*isolation & purification ; Spermatozoa/microbiology ; Staining and Labeling/*methods ; }, abstract = {Any hitherto available technique used to detect the presence of Wolbachia, a rickettsia-like endosymbiont of Drosophila simulans, i.e., crossing with tester strains, electron microscope observations, DAPI-staining of embryo, or amplification of specific sequences by PCR are notably time-consuming techniques. Staining sperm cysts with the fluorochrome DAPI is here shown to be a fast and reliable technique to detect the symbiont and evaluate the level of infection of individual males. Wolbachia cells are abundant in sperm cysts and make up a mass which is eliminated in the waste bag during spermatogenesis. Such a mass was absent in all those strains known to be aposymbiotic. In infected strains, tetracycline treatment led to the disappearance of the symbionts within sperm cysts. In aging males, a decrease in the proportion of infected cysts were observed. There was however no apparent change in the number of symbionts in the cysts which remained infected. The reduction of reproductive incompatibility, which is observed in older males, may be explained by this process, i.e., a decrease in the frequency of infected cysts.}, } @article {pmid8436273, year = {1993}, author = {Rigaud, T and Juchault, P}, title = {Conflict between feminizing sex ratio distorters and an autosomal masculinizing gene in the terrestrial isopod Armadillidium vulgare Latr.}, journal = {Genetics}, volume = {133}, number = {2}, pages = {247-252}, pmid = {8436273}, issn = {0016-6731}, mesh = {Animals ; Bacteria/genetics/isolation & purification ; Crosses, Genetic ; Crustacea/*genetics/microbiology ; Female ; Gene Frequency ; *Genes, Dominant ; Hermaphroditic Organisms ; Male ; Phenotype ; *Sex Determination Analysis ; Sex Determination Processes/genetics ; *Sex Ratio ; }, abstract = {Female sex determination in the pill bug Armadillidium vulgare is frequently under the control of feminizing parasitic sex factors (PSF). One of these PSF is an intracytoplasmic Wolbachia-like bacterium (F), while the other (f) is suspected of being an F-bacterial DNA sequence unstably integrated into the host genome. In most wild populations harboring PSF, all individuals are genetic males (ZZ), and female phenotypes occur only due to the presence of PSF which overrides the male determinant carried by the Z chromosome (females are thus ZZ +F or ZZ +f neo-females). Here we report the effects of the conflict between these PSF and a dominant autosomal masculinizing gene (M) on phenotypes. The M gene is able to override the feminizing effect of the f sex factor and, consequently, male sex may be restored. However, M is unable to restore male sex when competing with the F bacteria. It seems that the main effect of M is to delay the expression of F bacteria slightly, inducing intersex phenotypes. Most of these intersexes are functional females, able to transmit the masculinizing gene. The frequency of M and its effects on the sex ratio in wild populations are discussed.}, } @article {pmid9087560, year = {1993}, author = {Campbell, BC and Steffen-Campbell, JD and Werren, JH}, title = {Phylogeny of the Nasonia species complex (Hymenoptera: Pteromalidae) inferred from an internal transcribed spacer (ITS2) and 28S rDNA sequences.}, journal = {Insect molecular biology}, volume = {2}, number = {4}, pages = {225-237}, doi = {10.1111/j.1365-2583.1994.tb00142.x}, pmid = {9087560}, issn = {0962-1075}, mesh = {Animals ; Base Sequence ; *DNA, Ribosomal ; Molecular Sequence Data ; Phylogeny ; *RNA, Ribosomal, 28S ; Sequence Homology, Nucleic Acid ; Wasps/classification/*genetics ; }, abstract = {The phylogeny of the cryptic species complex of wasps in the genus Nasonia was inferred by analysis of nucleotide sequences of an rDNA internal transcribed spacer (ITS2) and the D2 region of 28S rDNA. Phylogenetic analysis showed that N. vitripennis descended from a theoretical common ancestor with that of a lineage that diverged into N. longicornis and N. giraulti. Differences in the ITS2 regions clearly distinguished two strains of N. giraulti. Another member of the Dibrachys Group, Trichomalopsis dubius, was placed outside of the Nasonia complex. The D2 region had a base substitution rate approximately 2 times slower than the ITS2 region and was used to resolve the phylogenetic affiliation of an eulophid, Melittobia digitata, to the pteromalids. Tree topology of the Nasonia complex was congruent with the phylogeny of a cluster of Wolbachia bacteria which infect these insects. The possible role of these bacteria in driving speciation is discussed.}, } @article {pmid8163156, year = {1993}, author = {Grandjean, F and Rigaud, T and Raimond, R and Juchault, P and Souty-Grosset, C}, title = {Mitochondrial DNA polymorphism and feminizing sex factors dynamics in a natural population of Armadillidium vulgare (Crustacea, Isopoda).}, journal = {Genetica}, volume = {92}, number = {1}, pages = {55-60}, pmid = {8163156}, issn = {0016-6707}, mesh = {Animals ; Crustacea/*genetics/microbiology ; DNA, Mitochondrial/*genetics ; *Extrachromosomal Inheritance ; Female ; Haplotypes ; *Hermaphroditic Organisms ; Male ; Models, Genetic ; *Polymorphism, Genetic ; Rickettsiaceae/isolation & purification/*physiology ; *Sex Determination Analysis ; *Sex Determination Processes ; Sex Ratio ; Symbiosis ; }, abstract = {Sex determination in Armadillidium vulgare may be under the control of two parasitic sex factors that reverse genetic males into functional neo-females. The first feminizing factor (F) is a Wolbachia and the other (f) is probably a sequence of the F bacterial DNA unstably integrated into the host genome. Both of these feminizing factors are mainly maternally transmitted. Here we investigate the mitochondrial DNA polymorphism of wild iso-female lineages harbouring either F or f. Among the four haplotypes present in the population, two were the f-harbouring lineages, while two were common to the F- and f-harbouring lineages. This result suggests that there has been an introgression of the f factor into lineages infected by F Wolbachia. Based on previous data, we propose two different ways to account for such introgression. Given the particular dynamics of feminizing factors (f-harbouring lineages increase in populations at the expense of F-harbouring lineages), such an introgression should prevent the replacement of F-linked mitochondrial types by f-linked mitochondrial types in wild populations.}, } @article {pmid8096432, year = {1993}, author = {Ndiaye, M and Mattei, X}, title = {Endosymbiotic relationship between a rickettsia-like microorganism and the male germ-cells of Culex tigripes.}, journal = {Journal of submicroscopic cytology and pathology}, volume = {25}, number = {1}, pages = {71-77}, pmid = {8096432}, issn = {1122-9497}, mesh = {Animals ; Culicidae/*parasitology ; Host-Parasite Interactions ; Male ; *Rickettsia/ultrastructure ; Rickettsia Infections/*transmission ; Seminal Vesicles/ultrastructure ; Spermatogonia/*parasitology/ultrastructure ; Symbiosis ; Testis/parasitology/ultrastructure ; }, abstract = {The male mosquitoes of the species Culex tigripes possess endosymbionts localized in the cytoplasm of the germ-cells. The somatic cells of the testis do not exhibit this particularity. The spermatogonia, spermatocytes and spermatids all possess a few symbionts. They are eliminated in the residual body at the end of spermiogenesis when the spermatozoon matures. They are absent from the seminal vesicles. A few spermatocytes and spermatids show a proliferation of the endosymbionts. This seems to be associated with an abnormal spermiogenesis. These symbionts are comparable to the Wolbachia described in two other species of mosquitoes: Culex pipiens and Aedes scutellaris.}, } @article {pmid1361987, year = {1992}, author = {Rousset, F and Bouchon, D and Pintureau, B and Juchault, P and Solignac, M}, title = {Wolbachia endosymbionts responsible for various alterations of sexuality in arthropods.}, journal = {Proceedings. Biological sciences}, volume = {250}, number = {1328}, pages = {91-98}, doi = {10.1098/rspb.1992.0135}, pmid = {1361987}, issn = {0962-8452}, mesh = {Animals ; Base Sequence ; DNA, Ribosomal/genetics/*isolation & purification ; Escherichia coli/genetics ; Female ; Insecta/genetics/*physiology ; Male ; Molecular Sequence Data ; Oligodeoxyribonucleotides ; Ovary ; Phylogeny ; Polymerase Chain Reaction ; Rickettsiaceae/*physiology ; Sequence Homology, Nucleic Acid ; *Sex Determination Analysis ; Testis ; Wasps/genetics/*physiology ; }, abstract = {Rickettsia-like maternally inherited bacteria have been shown to be involved in a variety of alterations of arthropod sexuality, such as female-biased sex ratios, parthenogenesis, and sterility of crosses either between infected males and uninfected females or between infected individuals (cytoplasmic incompatibility). We have characterized several of these microorganisms through partial sequences of the small (16S) and large (23S) subunit ribosomal DNA. All the symbionts identified, which include several cytoplasmic incompatibility microorganisms, several endosymbionts of terrestrial isopods, and symbionts of two thelytokous Trichogramma wasp species, belong to a monophyletic group of related symbionts, some of which have previously been detected in several insects exhibiting cytoplasmic incompatibility. Three molecular lineages can be identified on the basis of 16S as well as 23S sequences. Although they are only known as endocellular symbionts, Wolbachia spread by horizontal transfer across host lineages as evidenced by their diversification which occurred long after that of their hosts, and by the non-congruence of the phylogenetic relationships of symbionts and their hosts. Indeed, symbionts of two different lineages have been found in the same host species, whereas closely related endosymbionts are found in distinct insect orders. Isopod endosymbionts form a separate lineage, and they can determine feminization as well as cytoplasmic incompatibility. The ability to determine cytoplasmic incompatibility, found in all lineages, is probably ancestral to this group.}, } @article {pmid1557375, year = {1992}, author = {O'Neill, SL and Giordano, R and Colbert, AM and Karr, TL and Robertson, HM}, title = {16S rRNA phylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {89}, number = {7}, pages = {2699-2702}, pmid = {1557375}, issn = {0027-8424}, mesh = {Animals ; Bacteria/*genetics ; Bacterial Infections/diagnosis/veterinary ; Base Sequence ; Insecta/genetics/*microbiology ; Molecular Sequence Data ; Oligodeoxyribonucleotides/chemistry ; RNA, Ribosomal, 16S/*genetics ; Symbiosis ; }, abstract = {Bacterial endosymbionts of insects have long been implicated in the phenomenon of cytoplasmic incompatibility, in which certain crosses between symbiont-infected individuals lead to embryonic death or sex ratio distortion. The taxonomic position of these bacteria has, however, not been known with any certainty. Similarly, the relatedness of the bacteria infecting various insect hosts has been unclear. The inability to grow these bacteria on defined cell-free medium has been the major factor underlying these uncertainties. We circumvented this problem by selective PCR amplification and subsequent sequencing of the symbiont 16S rRNA genes directly from infected insect tissue. Maximum parsimony analysis of these sequences indicates that the symbionts belong in the alpha-subdivision of the Proteobacteria, where they are most closely related to the Rickettsia and their relatives. They are all closely related to each other and are assigned to the type species Wolbachia pipientis. Lack of congruence between the phylogeny of the symbionts and their insect hosts suggest that horizontal transfer of symbionts between insect species may occur. Comparison of the sequences for W. pipientis and for Wolbachia persica, an endosymbiont of ticks, shows that the genus Wolbachia is polyphyletic. A PCR assay based on 16S primers was designed for the detection of W. pipientis in insect tissue, and initial screening of insects indicates that cytoplasmic incompatibility may be a more general phenomenon in insects than is currently recognized.}, } @article {pmid1350096, year = {1992}, author = {Rousset, F and Vautrin, D and Solignac, M}, title = {Molecular identification of Wolbachia, the agent of cytoplasmic incompatibility in Drosophila simulans, and variability in relation with host mitochondrial types.}, journal = {Proceedings. Biological sciences}, volume = {247}, number = {1320}, pages = {163-168}, doi = {10.1098/rspb.1992.0023}, pmid = {1350096}, issn = {0962-8452}, mesh = {Animals ; Base Sequence ; Cytoplasm/microbiology ; DNA, Bacterial/genetics ; DNA, Mitochondrial/genetics ; Drosophila/genetics/*microbiology ; Female ; Male ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rickettsiaceae/*genetics/isolation & purification ; Sequence Homology, Nucleic Acid ; Symbiosis ; }, abstract = {Sequences of a segment of the 16S ribosomal DNA of Wolbachia, a rickettsia-like microorganism responsible for cytoplasmic incompatibility in Drosophila simulans, have been obtained after polymerase chain reaction (PCR) amplification. Their comparison with other eubacterial sequences allows us to assign these endosymbionts to the alpha subdivision of purple bacteria. Four related sequences have been obtained for microorganisms carried by eight isofemale lines representative of the three mitochondrial types of D. simulans. Their phylogeny and level of divergence do not parallel that of the mitochondrial DNA, suggesting that several independent infections occurred. There is no direct relation between bacterial phylogeny and formerly identified incompatibility types.}, } @article {pmid1371057, year = {1992}, author = {Fryer, JL and Lannan, CN and Giovannoni, SJ and Wood, ND}, title = {Piscirickettsia salmonis gen. nov., sp. nov., the causative agent of an epizootic disease in salmonid fishes.}, journal = {International journal of systematic bacteriology}, volume = {42}, number = {1}, pages = {120-126}, doi = {10.1099/00207713-42-1-120}, pmid = {1371057}, issn = {0020-7713}, mesh = {Animals ; Base Sequence ; Fish Diseases/*microbiology ; Molecular Sequence Data ; RNA, Bacterial/chemistry ; RNA, Ribosomal, 16S/chemistry ; Rickettsiaceae/classification/genetics/*isolation & purification ; Salmon/*microbiology ; }, abstract = {A novel intracellular pathogen morphologically similar to the ehrlichiae has been isolated in cell culture and identified as the cause of an epizootic disease of salmonid fish. Like the ehrlichiae, the salmonid pathogen, designated strain LF-89, replicates within membrane-bound cytoplasmic vacuoles in host cells. This agent is the first with characteristics of this type to be isolated from a fish. Analysis of the LF-89 16S rRNA indicated that, unlike the ehrlichiae, LF-89 is a gamma proteobacterium distantly related to Coxiella burnetii and perhaps Wolbachia persica. A new genus and species (Piscirickettsia salmonis gen. nov., sp. nov.) are proposed for this organism, with ATCC(R) VR 1361 as the type strain.}, } @article {pmid1743484, year = {1991}, author = {Montchamp-Moreau, C and Ferveur, JF and Jacques, M}, title = {Geographic distribution and inheritance of three cytoplasmic incompatibility types in Drosophila simulans.}, journal = {Genetics}, volume = {129}, number = {2}, pages = {399-407}, pmid = {1743484}, issn = {0016-6731}, mesh = {Animals ; Crosses, Genetic ; *Cytoplasm ; Drosophila/*genetics/microbiology ; Female ; Genetic Variation ; Male ; Reproduction ; Rickettsiaceae/isolation & purification ; Symbiosis ; }, abstract = {Wolbachia-like microorganisms have been implicated in unidirectional cytoplasmic incompatibility between strains of Drosophila simulans. Reduced egg eclosion occurs when females from uninfected strains (type W) are crossed with males from infected strains (type R). Here we characterize a third incompatibility type (type S) which is also correlated with the presence of Wolbachia-like microorganisms. Despite the fact that the symbionts cannot be morphologically distinguished, we observed complete bidirectional incompatibility between R and S strains. This indicates that the determinants of incompatibility are different in the two infected types. S/W incompatibility is unidirectional and similar to R/W incompatibility. A worldwide survey of D. simulans strains showed that type S incompatibility was found only in insular populations which harbor the mitochondrial type SiI. Both W and R types were found among mainland and island populations harboring the worldwide mitochondrial type SiII. Type S incompatibility could be involved in the reinforcement of the geographical isolation of SiI populations.}, } @article {pmid2076821, year = {1990}, author = {Hoffmann, AA and Turelli, M and Harshman, LG}, title = {Factors affecting the distribution of cytoplasmic incompatibility in Drosophila simulans.}, journal = {Genetics}, volume = {126}, number = {4}, pages = {933-948}, pmid = {2076821}, issn = {0016-6731}, mesh = {Aging ; Analysis of Variance ; Animals ; Cytoplasm/*physiology ; Drosophila/*genetics/physiology ; Female ; Male ; Oviposition ; *Polymorphism, Genetic ; Reproduction/genetics ; Spermatozoa/physiology ; Temperature ; }, abstract = {In Drosophila simulans a Wolbachia-like microorganism is responsible for reduced egg-hatch when infected males mate with uninfected females. Both incompatibility types have previously been found in North America, Europe and Africa. Some California populations have remained polymorphic for over two years, and the infection is apparently spreading in central California. Egg hatch proportions for wild-caught females from polymorphic populations show that the incompatibility system acts in nature, but egg mortality rates are apparently lower than observed in laboratory populations. Although infected females maintained under various laboratory conditions never produce uninfected offspring, some wild-caught infected females produce both infected and uninfected progeny. This helps explain the persistence of a low frequency of uninfected flies in predominantly infected populations and may also explain the other polymorphisms observed. Fitness comparisons of infected and uninfected stocks, including both larval and adult fitness components, indicate that fecundity may be the component most affected. Infected females suffer a fecundity reduction of 10-20% in the laboratory, but the reduction seems to be smaller in nature. A theoretical analysis provides some insight into the population biology of the infection.}, } @article {pmid2753854, year = {1989}, author = {Weisburg, WG and Dobson, ME and Samuel, JE and Dasch, GA and Mallavia, LP and Baca, O and Mandelco, L and Sechrest, JE and Weiss, E and Woese, CR}, title = {Phylogenetic diversity of the Rickettsiae.}, journal = {Journal of bacteriology}, volume = {171}, number = {8}, pages = {4202-4206}, pmid = {2753854}, issn = {0021-9193}, support = {AI 20190/AI/NIAID NIH HHS/United States ; AI 22910/AI/NIAID NIH HHS/United States ; }, mesh = {Bacteria/genetics ; Base Sequence ; Biological Evolution ; Molecular Sequence Data ; *Phylogeny ; RNA, Ribosomal/*genetics ; RNA, Ribosomal, 16S/*genetics ; Rickettsiaceae/*genetics ; Species Specificity ; }, abstract = {Small subunit rRNA sequences have been determined for representative strains of six species of the family Rickettsiaceae: Rickettsia rickettsii, Rickettsia prowazekii, Rickettsia typhi, Coxiella burnetii, Ehrlichia risticii, and Wolbachia persica. The relationships among these sequences and those of other eubacteria show that all members of the family Rickettsiaceae belong to the so-called purple bacterial phylum. The three representatives of the genus Rickettsia form a tight monophyletic cluster within the alpha subdivision of the purple bacteria. E. risticii also belongs to the alpha subdivision and shows a distant yet specific relationship to the genus Rickettsia. However, the family as a whole is not monophyletic, in that C. burnetii and W. persica are members of the gamma subdivision. The former appears to show a specific, but rather distant, relationship to the genus Legionella.}, } @article {pmid3409790, year = {1988}, author = {el Shoura, SM}, title = {Fine structure of the vagina, accessory glands, uterus, oviducts and Gene's organ in the unfed tick, Ornithodoros (Pavlovskyella) erraticus (Ixodoidea: Argasidae).}, journal = {Experimental & applied acarology}, volume = {4}, number = {2}, pages = {95-108}, pmid = {3409790}, issn = {0168-8162}, mesh = {Animals ; Arachnid Vectors/*ultrastructure ; Female ; Food Deprivation ; Genitalia, Female/ultrastructure ; Microscopy, Electron ; Oviducts/ultrastructure ; Ticks/*ultrastructure ; Uterus/ultrastructure ; Vagina/ultrastructure ; }, abstract = {The fine structure of the reproductive system of the unfed female Ornithodoros (Pavloskyella) erraticus is described. The vagina consisting of vestibular (VV) and cervical (CV) regions is formed of a single epithelial layer lined with cuticle and surrounded by muscle layers. Epithelial cells of VV show no signs of activity, while those of CV have structural features of transporting epithelia. A pair of tubular accessory glands opening at the junction of the two vaginal regions consist of a layer of microvillate columnar cells containing rough endoplasmic reticulum; this is possibly involved in the formation of colloidal material which presumably coats the egg surface during its passage through the vagina. The bilobed uterus opening into the cervical vagina secretes material that is possibly responsible for the release of sperm from the endospermatophore. The paired oviducts joining the uterine lobes anteriorly have distinctive distal (DO) and proximal (PO) parts as well as 'ampullae' (AMP) at their junction. DO cells contain many lysosome-like structures which probably play a role in breaking down some sperms ascending to the ovary. Cells of AMP are packed with rickettsia-like Wolbachia, while those of PO contain a few granules. Gene's organ, present in female ticks only, consists of a stalk and two horns of epithelium lined internally by a cuticular layer. Bundles of microtubules are detected in the horn cell cytoplasm. This organ functions to waterproof the eggs with a waxy layer just before they are deposited.}, } @article {pmid6145350, year = {1984}, author = {Avakyan, AA and Popov, VL}, title = {Rickettsiaceae and Chlamydiaceae: comparative electron microscopic studies.}, journal = {Acta virologica}, volume = {28}, number = {2}, pages = {159-173}, pmid = {6145350}, issn = {0001-723X}, mesh = {Chlamydia/cytology/growth & development/ultrastructure ; Chlamydiaceae/classification/*ultrastructure ; Microscopy, Electron ; Models, Structural ; Rickettsia/cytology/ultrastructure ; Rickettsiaceae/classification/*ultrastructure ; Species Specificity ; }, abstract = {The structure and cytopathology of obligate intracellular bacteria belonging to families Rickettsiaceae and Chlamydiaceae and their interaction with eukaryotic host cells were compared in electron microscopic studies. "Rickettsia-like" and "chlamydia-like" types of organization of bacterial cells and their interaction with host cells are presented. The rickettsia-like type is characterized by short rod-shaped cells multiplying freely (extravacuolarly) in the cytoplasm or nucleoplasm of the host cell; the chlamydia-like type has spherical cells multiplying inside the cytoplasmic vacuole limited by the host membrane. The rickettsia-like type includes the genus Rickettsia and rod-shaped symbionts from genera Wolbachia and Symbiotes ; the chlamydia-like type falls into genera Chlamydia, Ehrlichia, Cowdria and Neorickettsia . The transitional types represented by Wolbachia persica (type 1), Coxiella and Rickettsiella (type 2) are also described. The possible evolutional relationships of the genera comprising both families are considered and their classification is proposed.}, } @article {pmid6863975, year = {1983}, author = {Larsson, R}, title = {A rickettsia-like microorganism similar to Wolbachia pipientis and its occurrence in Culex mosquitoes.}, journal = {Journal of invertebrate pathology}, volume = {41}, number = {3}, pages = {387-390}, doi = {10.1016/0022-2011(83)90258-6}, pmid = {6863975}, issn = {0022-2011}, mesh = {Animals ; Culex/*microbiology ; Female ; Rickettsiaceae/*isolation & purification/ultrastructure ; }, } @article {pmid6404990, year = {1983}, author = {Awahmukalah, DS and Brooks, MA}, title = {Reproduction of an inbred strain of Culex pipiens prevented by loss of Wolbachia pipientis.}, journal = {Journal of invertebrate pathology}, volume = {41}, number = {2}, pages = {184-190}, doi = {10.1016/0022-2011(83)90218-5}, pmid = {6404990}, issn = {0022-2011}, support = {AI 09914/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culex/microbiology/*physiology ; Female ; Male ; Reproduction ; Rickettsiaceae/*physiology ; }, } @article {pmid7411685, year = {1980}, author = {Wright, JD and Barr, AR}, title = {The ultrastructure and symbiotic relationships of Wolbachia of mosquitoes of the Aedes scutellaris group.}, journal = {Journal of ultrastructure research}, volume = {72}, number = {1}, pages = {52-64}, doi = {10.1016/s0022-5320(80)90135-5}, pmid = {7411685}, issn = {0022-5320}, support = {AI-11847/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology/ultrastructure ; Animals ; Endoplasmic Reticulum/ultrastructure ; Female ; Oocytes/microbiology ; Ovary/microbiology ; Rickettsiaceae/physiology/*ultrastructure ; Symbiosis ; }, } @article {pmid671578, year = {1978}, author = {Wright, JD and Sjöstrand, FS and Portaro, JK and Barr, AR}, title = {The ultrastructure of the rickettsia-like microorganism Wolbachia pipientis and associated virus-like bodies in the mosquito Culex pipiens.}, journal = {Journal of ultrastructure research}, volume = {63}, number = {1}, pages = {79-85}, doi = {10.1016/s0022-5320(78)80046-x}, pmid = {671578}, issn = {0022-5320}, mesh = {Animals ; Bacteriophages/*ultrastructure ; Cell Division ; Cell Membrane/ultrastructure ; Cell Nucleus/ultrastructure ; Culex/*microbiology ; Ribosomes/ultrastructure ; Rickettsiaceae/*ultrastructure ; }, } @article {pmid600395, year = {1977}, author = {Sixl-Voigt, B and Roshdy, MA and Nosek, J and Sixl, W}, title = {Electronmicroscopic investigations of Wolbachia-like microorganisms in Haemaphysalis inermis.}, journal = {Mikroskopie}, volume = {33}, number = {9-10}, pages = {255-257}, pmid = {600395}, issn = {0026-3702}, mesh = {Animals ; Female ; Malpighian Tubules/microbiology ; Nymph/microbiology ; Ovary/microbiology ; Rickettsiaceae/*ultrastructure ; Ticks/*microbiology ; }, } @article {pmid241251, year = {1975}, author = {Cory, J and Yunker, CE and Howarth, JA and Hokama, Y and Hughes, LE and Thomas, LA and Clifford, CM}, title = {Isolation of spotted fever group and Wolbachia-like agents from field-collected materials by means of plaque formation in mammalian and mosquito cells.}, journal = {Acta virologica}, volume = {19}, number = {5}, pages = {443-445}, pmid = {241251}, issn = {0001-723X}, mesh = {Aedes ; Animals ; *Cell Line ; Dermacentor/microbiology ; Haplorhini ; Kidney ; Moths ; Rickettsiaceae/*isolation & purification ; Sciuridae/microbiology ; *Viral Plaque Assay ; }, abstract = {Three isolations from ticks (Dermacentor occidentalis) of a rickettsia of the spotted fever group and 5 isolations from chipmunk (Eutamias rugicaudus) blood of a Wolbachia-like agent were obtained from plaques formed in Singh's Aedes albopictus (mosquito) and Vero (African green monkey kidney) cell cultures. These organisms could not be isolated by injection of the infected ticks or blood into embryonated chicken eggs, guinea pigs, or voles (Microtus pennsylvanicus), but fluid cultures of Grace's Antheraea eucalypti (moth1 and Singh's A. albopictus cells inoculated with the bloods yielded the Wolbachia-like agent.}, } @article {pmid808618, year = {1975}, author = {Portaro, JK and Barr, AR}, title = {"Curing" Wolbachia infections in Culex pipiens.}, journal = {Journal of medical entomology}, volume = {12}, number = {2}, pages = {265}, doi = {10.1093/jmedent/12.2.265}, pmid = {808618}, issn = {0022-2585}, mesh = {Animals ; Culex/*microbiology ; Female ; Male ; *Rickettsia ; Rickettsiaceae ; }, } @article {pmid53109, year = {1975}, author = {Croizier, G and Meynadier, G and Morel, G and Capponi, M}, title = {[Immunologic comparison between some Wolbachiae and research of antigentic community with other members of the order of Rickettsiales].}, journal = {Bulletin de la Societe de pathologie exotique et de ses filiales}, volume = {68}, number = {2}, pages = {133-141}, pmid = {53109}, mesh = {Antibodies, Bacterial/analysis ; Antigens, Bacterial/*analysis ; Arthropods/microbiology ; Epitopes ; Rickettsia/immunology ; Rickettsiaceae/*immunology ; }, abstract = {The antigenic relationship between some rickettsiae of arthropods, which do not transmit germs pathogenic to vertebrates was investigated by agglutination and immunofluorescence techniques. The comparison of R. melolonthae, R. tipulae, R. cetonidarum, R. grylli and of the rickettsiae of the scorpion Buthus occitanus shows that the Wolbachiae are divided into three serological groups. The insect crystallogen rickettsiae belong to the first group, R. grylli and the scorpion Rickettsia are antigenically distinct (tables I and II). No antigenic relationship was found between these arthropod microorganisms and the following agents of vertebrate rickettsioses and chlamydioses : historical typhus, murine typhus, pustulous fever, Q-fever, epizootic absortion of sheep and psittacosis (tables III and IV). Bacteria of OX2 and OX19 strain of P. vulgaris when mixed with anti-Wolbachiae serums of I, II, III groups, give a negative Weil a Felix reaction. The OXK Proteus are however agglutinated by 4 out of 6 rabbit anti-Rickettsiella serums of the group. This fact calls for a further investigation regarding the comparison of these Rickettsiella with various strains of R. tsutsugamushi.}, } @article {pmid20018, year = {1975}, author = {Yen, JH}, title = {Transovarial transmission of Rickettsia-like microorganisms in mosquitoes.}, journal = {Annals of the New York Academy of Sciences}, volume = {266}, number = {}, pages = {152-161}, doi = {10.1111/j.1749-6632.1975.tb35096.x}, pmid = {20018}, issn = {0077-8923}, mesh = {Aedes/microbiology/physiology ; Animals ; Crosses, Genetic ; Culex/microbiology/physiology ; Culicidae/*microbiology ; Female ; Insect Vectors/*microbiology ; Male ; Ovary/microbiology/ultrastructure ; Ovum/microbiology ; *Rickettsiaceae/growth & development/ultrastructure ; Testis/microbiology/ultrastructure ; }, abstract = {The wolbachiae found in Culex pipiens and the Tafahi strain of the A. scutellaris group are small rickettsia-like symbionts of the gonads. They are extrachromosomal self-replicating units that are vertically transmitted through the ovaries. Their presence in the only two groups of mosquitoes known to exhibit incompatibility, the fact that they are found in only the Tafahi strain, and the loss of incompatibility after removal of Wolbachia in C. pipiens are compelling evidence for the role that Wolbachia plays in incompatibility.}, } @article {pmid4997776, year = {1971}, author = {Devauchelle, G and Vago, C and Meynadier, G}, title = {[Comparative ultrastructure of Ricketsiella melolonthae (Rickettsiales Wolbachiae) and of the agent of venereal lymphogranulomatosis (Rickettsiales Clamydiaceae)].}, journal = {Comptes rendus hebdomadaires des seances de l'Academie des sciences. Serie D: Sciences naturelles}, volume = {272}, number = {23}, pages = {2972-2974}, pmid = {4997776}, mesh = {Escherichia coli/physiology ; HeLa Cells ; Lymphogranuloma Venereum/microbiology ; Microscopy, Electron ; Rickettsia/classification/*cytology/growth & development ; }, }